A Conversation With Christine Lagarde

https://www.cfr.org/event/conversation-christine-lagarde Past Event — December 12, 2018 8:30am EST Issei Kato/Reuters Christine Lagarde Managing Director, International Monetary Fund Presider James Manyika Chairman and Director, McKinsey Global Institute, and Senior Partner, McKinsey & Company, Inc.; Member, Board of Directors, Council on Foreign Relations Introductory Remarks Rachel Vogelstein Douglas Dillon Senior Fellow and Director of the Women and Foreign Policy Program, Council on Foreign Relations from Women and Foreign Policy Program Despite the positive relationship between women’s labor force participation and GDP growth, legal barriers undermine female economic potential in every region of the world. Gender-based legal restrictions exist in almost 90% of nations, ranging from limitations on property ownership to prohibitions on signing contracts. Christine Lagarde discusses the economic implications of gender inequality under the law and outlines policy recommendations to accelerate women’s economic participation. For more information, visit the Women's Workplace Equality Index. VOGELSTEIN: Good morning. Welcome to the Council on Foreign Relations. My name is Rachel Vogelstein. I lead the Women and Foreign Policy Program here at CFR, which analyzes how elevating the status of women and girls advances U.S. foreign-policy objectives. On behalf of Richard Haass, the president of the Council, it is my great pleasure to commence our first CFR Symposium on Women and the Law. I want to begin by extending my gratitude to our esteemed speakers for joining us this morning. I also want to thank all of you for participating today, and welcome everyone tuning in to our live cast. And I’d like to extend a special appreciation to those of you who traveled here from out of town. We are very glad to have you with us. Today’s Symposium on Women and the Law is focused on an issue that has been part of our national and global dialogue for well over a year—how to level the economic playing field for women. In capitals and boardrooms, in the workplace and on the streets, through hashtags and social media, even around the proverbial water cooler and dinner table, good men and women alike have grappled with the persistent barriers that hamper women in the workplace in the 21st century, as well as the question of how to address them. To help answer this question, scholars in the Women and Foreign Policy Program here at the Council have created several new tools to assess the status of women in the workplace around the world and explain why leveling the legal playing field for women is so critical to economic prosperity and stability. To that end, today we are launching the Women’s Workplace Equality Index, which is the first-ever global ranking of countries based on gender equality in the workplace. Under CFR’s gender-equality ranking, which draws upon World Bank data, Australia comes in first. Canada is second. Mexico is fifth. And notably, the United States is not even in the top ten, falling at 20th. At the bottom of the list are Iran, Sudan, Qatar, Syria, and Yemen. The index highlights the pervasive nature of legal barriers to women in the workplace, finding that over one hundred countries restrict the kinds of jobs that women can hold, that fifty-nine countries lack any legal protection against sexual harassment in the workplace whatsoever, and not a single nation of 189 covered has a level legal playing field for women at work. Today we are also issuing a new report on Women and the Law, which analyzes the five areas in which the greatest obstacles to women’s economic participation endure: financial inclusion, national identification law, land rights, workplace discrimination, and family law. This report reflects field research conducted by CFR scholars in Tanzania, Nigeria, and Pakistan. And it covers many of the themes we’ll discuss in our panel sessions today. So our data confirm that legal barriers to women’s workforce participation persist everywhere in the world. But why do these barriers to women’s equality in the workplace matter? Our new interactive report, entitled Growing Economies Through Gender Parity, outlines the economic states, visualizing data from the McKinsey Global Institute showing that closing the gender gap in workforce participation could add a staggering $28 trillion to global GDP. Both advanced and developing economies alike stand to benefit if women are able to participate in the labor force to the same extent as men. The U.S. economy could grow by 19 percent, China’s by 20 percent, Mexico’s by 43 percent, and India’s by a remarkable 60 percent. You can find each of these new reports in the materials at your seats and online at CFR.org. The bottom line from our research is this: In the 21st century, nations cannot get ahead economically by leaving half of their population behind. Leveling the playing field affords an opportunity to take stock of the barriers that persist for women at work, to explore policy approaches in both the public and private sectors, and to ask hard questions about the way forward. And we have four terrific panels to accomplish just that. Our keynote session will feature a conversation with IMF Managing Director Christine Lagarde to explore the economic implications of gender equality under the law. Our second session, with World Bank CEO Kristalina Georgieva, will analyze financial inclusion in the digital age. Our third session will examine the issue of workplace discrimination in the era of an increasingly global #MeToo movement. And our final session will assess efforts to improve women’s economic participation through family-law reform, perhaps the most contentious area of the law for women. We have a stellar roster of panelists, and we’re grateful to each of them for sharing their expertise with us. Before we begin, a special word of thanks goes to the Bill and Melinda Gates Foundation, which has generously sponsored today’s symposium and our work on women and economic growth. In particular, I want to thank Rosita Najmi and her colleagues for their leadership and continued support for the Council’s work. I’d also like to thank my Women and Foreign Policy Program colleagues, whose scholarship has paved the way for our discussion today: Senior fellows Jamille Bigio, who led development of our new Workplace Equality Index; Meighan Stone, who is studying the global implications of the #MeToo movement; and Gayle Lemmon, Catherine Powell, and Carrie Bettinger-Lopez, who contributed to the materials that we’re releasing today. I’d also like to thank the extraordinary CFR team for making this symposium possible, including Stacey LaFollette, Kayla Ermanni, and Carrie Bueche, as well as the outstanding Rebecca Turkington, Alexandra Bro, and Rebecca Hughes of the Women and Foreign Policy team. Finally, before we get started, I’d like to remind everyone that today’s conversation will be on the record. With that, please join me in welcoming to the stage the honorable Christine Lagarde and James Manyika. Thank you very much. (Applause.) MANYIKA: Well, good morning, and welcome to this keynote session of the Symposium on Women and the Law. And the topic for this discussion is Leveling the Economic Playing Field. I’m James Manyika. I’m the chairman of the McKinsey Global Institute and also a member of the board of the Council on Foreign Relations. And it’s my distinct honor—and I think it’s quite fitting that we have Christine Lagarde here to discuss this. I think it’s worth—I mean, she’s obviously very well known to all of you, but I think it’s worth noting that you were the first woman managing director of the International Monetary Fund. And before that, she was also the finance and economic minister in France for many years. So it is really a real pleasure to have this conversation with you. I thought where we might start is to actually take an economic view to all these questions. And so when we talk about an uneven economic playing field, what are we talking about? What are the elements of inequity or a lack of parity when it comes to women and the economy? LAGARDE: Thank you, James. And good morning to all of you. One thing that you missed out in my resume, which is actually of interest to that room, is that I was also for many, many years a lawyer—(laughter)—and very much part of that community. I was also in my firm—at Baker McKenzie, I was the first managing partner. So I want to disclaimer first. Although head of the International Monetary Fund and surrounded by a whole bunch of terribly talented economists, a lawyer is always a lawyer. (Laughter.) So that’s one. Number two, if I may, how many of you in this room are lawyers? OK, that’s a pretty sizable number. How many in this room are partners in their respective law firm? MANYIKA: Two. Wow. I just see two hands. LAGARDE: OK. How many are managing partners in their respective law firms? OK, I rest my case. (Laughter.) I hope things can continue to change as they had for a period of time. But I’m afraid—I looked at numbers from the ABA and the IBA, thanks to my general counsel, who is a woman. Rhoda, thank you. And the numbers didn’t look that great. And there is a bit of backtracking in the profession that I’m a little bit worried about, because—and I come to your question—there is obviously a moral-philosophical case to be had concerning women’s access, parity, equality of opportunity, income, and blah, blah, blah, blah, blah. And I’m the daughter of the—(inaudible)—civilization, and I could plead that case. But I’m not going to. I just want to look at the economic data that we have. And whether you look—and thank you to the Council on Foreign Relationships (sic). You did a fabulous job. And I’m very pleased that under your leadership we are looking at those issues. There is an obvious case for making sure that women have equal access to education—primary, secondary, tertiary—equal access to the job market, equal compensation for equal jobs that they do. And that is reflected in numbers. It’s very, very clear. Let me give you—because many of your numbers are right, so I’m not going to repeat them. They—first of all, I think there is a distinction between advanced economies, generally put together under the OECD countries, and then you have the middle-income countries and you have the low-income countries. And whether you measure the female labor force participation or whether you measure the compensation, in all three instances of countries you have a significant gap. So women access the labor market less so than men. Women receive less compensation than men. And that is true across the board, in advanced, in middle-income, and in low-income countries. It is actually more so the case in middle-income countries. So the gap is, on average, in the OECD countries at about 16 percent when it comes to female labor force participation. It’s much higher than that, around 26 percent, in the middle-income countries. And it’s not that high in low-income countries, but what we measure there is uncertain because there is a mass of informal and informality in their economy, which makes the calculation a bit difficult. And that is not to mention the unreported, unaccounted-for, and yet terribly important work that is discharged generally by women. And that has to do with work at home; work to carry the water, literally. You know, I’m not just saying carrying the water, but literally doing that. So the gap is very significant. If you look at compensation now, on a global basis—so you take into account all countries and, you know, the entire population, working population—you’re talking about a difference of compensation of 50 percent—five-zero. That is huge. Now, you will say 50 percent. What is she talking about? Fifty percent on a global basis. If you look then at advanced economies, it’s much lower than that. I have to look at my numbers, not to give you any wrong ones in case you want to report it, but it is 16 percent. So, sorry, in F—female labor force participation, OECD countries, 14 percent; and in compensation, 16 percent. Those are the differences that we’re talking about. Now, does it matter? Well, yes, it does matter. It does matter enormously, because I think if you—in a way, the worst-case example, where there is a gender, female labor force participation, gap of 30 percent, assume—and all that is by way of modelization. It’s not real-life, you know, empirical evidence. It’s modelization because that’s what we’ve got to play with. But if you close that gap entirely and if you have the same participation in the labor force for both men and women, the overall GDP of that community, a nation in that particular case, would increase by 25 percent. So close the gap of 30 percent, equal 25 percent increase in the GDP. I have yet to meet a head of state, a head of government, or a finance minister—most of them will be men, don’t worry—who say to me, I am not interested in additional growth. I am not interested in making the cake a bit larger so that I can allocate it a bit better. So there is an obvious economic argument in increasing the size of the economy in order to then determine what policies will be applicable. So that’s really obvious. MANYIKA: Let’s— LAGARDE: And, by the way—one more thing—(laughter)—sorry—because that’s not the only thing it does. Many of you have heard the debate, particularly in light of what’s happening in France at the moment, about excessive and rising inequalities. The fact that you close those gaps, both in terms of female labor force participation and in terms of compensation, mathematically actually reduces the inequalities. That’s number one. Number two, observations that we’ve made, looking at the 189 membership that we have in the IMF, is that it also leads to a diversification of the economies. So on those two accounts, as well—because we know that non-diversified economies are very vulnerable. If you only depend on one single big—say, oil, for instance, or copper or, you know, some of those commodities—if you don’t diversify, if you don’t move into services promptly, you’re going to be at risk. So bringing women, reducing those two gaps, actually leads to that. MANYIKA: Well, you talk quite a bit about labor force participation rates and the inequity and lack of parity there, but you’ve also—in other settings I’ve heard you talk about other aspects of inequality, things like access to capital, access to technology, things that enable women to participate more fully in the economy. Say a little bit more about that. How important are those other economic access inequities? LAGARDE: Well, I’ll say a little bit, but I don’t want to say too much because my friend Kristalina is going to talk about financial inclusion, and on that subject I want to leave the ground to her, and they’ve done superb work in that respect. I’ll just say one thing because it’s a special research paper that we recently published which tries to examine what the impact of the future of work—understood as a very broad concept of impact of technologies at large, from artificial intelligence, to data mining, to robotization, and on, and you put it all together—and the compounded effect of that is bound to transform jobs, to remove some jobs, to affect the way in which is discharged and services are provided. If you try to anticipate the impact that it will have on a gender-desegregated basis, you realize very quickly that women are going to be more impacted than men and by significant account because you realize—and I know that McKinsey has done some great studies on that as well, but you realize that 11 percent of female jobs around the world are going to be significantly affected, if not removed, when 7 percent of men’s jobs will be equally—either affected significantly or just removed. So that’s a major change. How many jobs does it mean—away from percentages? It means 28 million female jobs in those thirty countries that are studied in that particular study. If you then extrapolate to the entire membership, you are talking about 280 million jobs that are either significantly affected, cannot be discharged without massive training and adjustment, or you are talking about jobs gone. Now people will say, why is that? Are women that stupid? No, don’t need to actually argue that point, but it’s simply because in many of those economies, women tend to discharge those routine, repetitive tasks that are much more easily automated or substituted by the new technologies that are coming to the markets—that’s all there is about it—which calls for policies. MANYIKA: Well, let’s come—I mean, you made a very compelling argument and case for why this makes economic sense, why we should have more parity and participation and so forth. Why do you think it’s so hard? Because when I look at countries that do need economic growth, whether it’s countries in Europe or even the United States, developed, developing economies, surely there should be enough momentum to do something about this. What are the barriers, do you think? Why is this so difficult? LAGARDE: I’ll just—this is a personal assessment of mine. I think that what we are seeing in terms of women’s role in our economies is not less than revolutionary, and revolutions are difficult to swallow. Now I’ll say two things. One is—having many lawyers in the room I want to make that point. There are multiple legal barriers to the inclusion of women on a parity basis, and again, tribute to the World Bank. The World Bank every two years produces a study of the legal discriminations that apply around our respective memberships. And we’ve done some sort of digging into those discriminations. Eighty-eight percent of countries around the world include in their constitution, in their law, in their executive orders, discriminations against women. OK, once you have said that you have said not much because you need to dig deep into what kind of discriminations are we talking about. So you have the combination of constitutional principles—whether you are talking about civil rights, whether you are talking about right to ownership, whether you are talking about—you then go into family law as well. Inheritance is a huge, big package of discrimination. And I want to celebrate here for second Tunisia because Tunisia, two weeks ago, has introduced in parliament a law that would equalize the status of men and women in case of passing away of the parents so that brothers and sisters can inherit equally. This is fiercely debated by the Islamist party in Tunisia on the basis that a man equals two women and therefore the inheritance should be appropriately and proportionately differentiated. So you have in many developing countries, particularly in sub-Saharan Africa—an interesting phenomenon actually from a legal point of view—which is that many of the old Spanish or French civil codes or soft laws as a result—there were some decrees that were taken at the time of colonizations and sometimes decolonization as well—which actually introduced and have maintained in those legislations discriminations against women. A decree from 1954 that was put in place in many of the ex-French colonies actually still to this day requires that the husband consent to the opening of a bank account for a woman. It seems like—well, I remember when I was a little girl—it doesn’t make me any younger, but anyway—(laughter)—that’s OK—my mother had to do that. When she wanted to open a bank account, my father had to consent, and for a few years, I could see them both signing the check because they had—there were restrictions in France to prevent that. So, bottom line, there are multiple legal discriminations around the globe on all those issues—economic access, inheritance—and you can understand the consequences. If a woman is not entitled to either own property or to inherit sufficiently, then she doesn’t have the collaterals that will be needed in order to secure a loan assuming she is entitled to access to a loan. So that really matters. And it—when it changes—when it changes it makes a difference. We did a study back in 2015. We haven’t redone it on the basis of the latest figures of Kristalina, but in 2015 we looked at all those countries that had those restrictions on the past, and we tried to see whether there was a difference as a result of a change. And in 50 percent of those countries that actually implemented those changes, we realized very quickly that there was within five years of the constitutional, or soft law, or discriminatory legal basis—there was a difference in female labor force participation. That is the case very clearly in countries like Namibia, in Peru, in Malawi, and various others. MANYIKA: But I think it’s quite easy to paint this as a challenge for developing economies. You still have legal restrictions, even in advanced economies like the United States and others, even in the tax code. Talk a little bit more about some of the challenges that you see in some of developed advanced economies like the United States. LAGARDE: Well, in the tax code there is one that is just in our face which actually applies to countries like Germany or France or, you know, a few others, which is that a tax—a tax unit where you assess whatever, you know, personal income tax you want to assess—a tax unit comprises a couple. So taxpayers are not individuals; they are a household comprising the two spouses. And guess what? Marginally most affected is going to be the, quote, unquote, secondary earner, and that is invariably the woman. So just by changing that and by turning the tax unit into a person rather than a family unit would actually eliminate that implied discrimination for those households that actually pay taxes. MANYIKA: Right. You know, well, one of the things that I found quite fascinating about a lot of the studies that you’ve done and that the IMF has done is to show that it’s not just about parity; in fact, the diversity in itself is also valuable when you think about the diverse skills, and profiles, and capabilities that diverse teams bring to—not just to companies, but also to the economy. And I’m quite curious about you think about that—that it’s not just about getting to parity, but leveraging the diversity of teams and organizations as well. LAGARDE: I’m not sure that we have actually done some research paper on that, but it’s—you know, in a way, that’s my life, so I don’t find it surprising because I—you know, I grew up with Baker McKenzie which was, at the time, the most diverse firm you could ever think, where, you know, equal right to all partners whether you sat in Rome, or New York, or in Beijing. And the IMF is Babylon. I mean, you have all languages, all nationalities, and we try to enforce diversity. And we have targets by gender, by countries, by groups of countries, by education—by educational background. And we measure our performance against those thresholds. And I’m not claiming that, oh, we’ve made it; we are at full parity. No, we still have a way to go. And I don’t—I would never trumpet success in that respect. Can I come back to one topic, because— MANYIKA: Yeah. LAGARDE: —now that you made me think about the latest research. There is one interesting one which I think would be of concern to some. We very—I very often hear the case that if you promote women, if you give them equal access and if you push the labor force participation using quotas, targets, segmented targets, and all the rest of it, it’s going to discriminate against men, OK. Now, we did—and actually it was driven by one of our men researchers, but we did a study on the actual economic impact of reducing the female labor force participation, of which those numbers that I’ve mentioned here result. But he went further into that to actually try to identify the female factor. And I know that there are two schools of thoughts. And I stand on the side of those who think that each gender actually brings characteristics, if not specific, and maybe not naturally driven, but characteristics that are gender-determined. That’s where I stand. I know there is the other current, and I respect them, and we can have a debate forever about it. But his study was, I think, premised on that basis. And the study really tried to identify what was the economic impact of that—those particularly gender-determined characteristics. And the conclusions of that study are that there is an added impact, because when you add an additional full-time equivalent, irrespective of gender, you get an up. If you add a full-time equivalent that is gender-distinct and begins to close the gap, you bring added value that respect in increased productivity. And that increased productivity—everything being normal in the economy, at least—brings added revenues, not just for those that raise the productivity, but to all genders. So as a result of that, men as well as women typically receive higher wages. That’s assuming economic rules operate. Then you have all the other principles of stakeholders and how much you distribute to your shareholders and how much you keep within and reallocate. That’s another debate. But on pure economics, increased productivity, brought about by the characteristic determined by gender, benefit both men and women. So to those who say, oh, men will be disadvantaged, no, not true. MANYIKA: Yeah. No, I actually found that research fascinating because it looked, I think, at the—you know, the elasticity of substitution and the— LAGARDE: Yeah. I tried to stay away from those words, because they are so—(laughter)— MANYIKA: Right, which is fascinating. LAGARDE: —for lawyers. MANYIKA: But I think you also found—one of the other things that I found interesting was the observation that, in fact, as the economies move towards more services-oriented economies, that also—that reallocation of sectors also benefits the economy. And women tend to be much, much more—the labor-participation rates for women in the services sector tend to be much, much higher. LAGARDE: True. MANYIKA: But I’m curious. I mean, beyond what you’re doing in the IMF itself as an organization, I’m just curious, as somebody who spends a lot of time in policy conversations and trying to influence and help people improve their economies, what are you doing as the IMF—as you talk to world leaders or business leaders, what are you doing to try and help address some of these challenges? LAGARDE: Well, first of all, it’s a little bit new in the organization. And it took me a few years to convince the board of the IMF that actually gender economics could be macro-critical. It’s only 50 percent of the population I’m talking about. (Laughter.) But it took a while. The World Bank was much better at doing it and is excellent at focusing on those issues. And I remember vividly the annual meeting where Bob Zoellick actually had women in various corners of these annual meetings to showcase the voices of women. So at the IMF it’s not that frequent. I think we are at a stage where it’s now really regarded as macro-critical. And we’ve introduced it at three levels, and I’ll go through those three. One is—because we do—we have three business lines. We do macroeconomic surveillance, so we audit economies around the world. That’s consented by all countries. It’s part of the contract. Second, we do technical assistance. And third, we do financial support. We never give grants. We give short-term loans in order to take countries out of their misery when they have balance-of-payment major crisis. So those are the three lines. If you look at those three, in surveillance it’s not part of the normal surveillance process. We piloted that through 39 countries. It’s now streamlined; mainstreamed, rather. And wherever it’s macro-critically relevant—in other words, we’re not going to do it in Norway, in Denmark, in Finland, because we have to use our resources where it’s going to be relevant and make the case—but in other countries we’re going to have that as part of the surveillance, and we’re going to look at the economic impact of discrimination, lack of access to finance, inability of women to do this, that, and the other. That’s number one. And we do give those—for instance, the tax advice that I’ve just mentioned, it was in the 2018 audit that we did of Germany. Change your tax system. In the 2018 audit of the U.S. economy, we said it’s maybe about time to have a real maternity leave, because you are one of the outliers in the OECD countries with no federal maternity leave, so to speak. So that’s what we do there. Technical assistance. We now provide a lot of what we call gender budgeting technical assistance so that countries, when they put their budget in place, instead of doing that blindly, they do it thinking, OK, is this particular fiscal policy going to help or hinder women? And as a result, if there is a determination within that government to actually give access to women, then focus on what will have positive impact. And the third one is whenever we have a lending arrangement with a country that is going through a lot of trouble caused by balance-of-payment crisis, either for internal or external reasons, we say when it’s macro-critically relevant, let’s examine what you could do specifically to help women access to the workforce. So we’ve done that in the case of Jordan, for instance, which is determined to help women access and have a better chance. We did that in the case of Niger. We did that in the case of Egypt. We are doing that in the case of Argentina. And it takes very different forms. As you can imagine with those countries, it often takes the form of having some child-care budget and improving the child-care facilities. But it takes unusual forms as well. In Jordan, for instance, it was a determination to actually spend more money on transportation, because unsafe transportations actually prevented many women from going from home to work because of fear what would happen in between. So those are the kinds of things that we try to do. MANYIKA: And do you—I mean, there was a point, as was pointed out at the beginning, that, in fact, we’ve been having societally this conversation now for about a year and a half or so. But as you think about the economic landscape, do you think this is a turning point? Do you— LAGARDE: No. No. No. I think it—I think it’s—we are on a road. We’ve come a long way. You know, I remember my mother co-signing checks with my father, bless both of them. But it’s an ongoing journey and one where we have to just continue to stand together shoulder to shoulder with people like you and many other men who believe that it’s actually worth it, and it’s not going to jeopardize our respective relationships. It’s a continuous road, because there is always a tendency to backtrack. There are always forces that will tend to suppress and repress rather than open barriers. MANYIKA: Open them. LAGARDE: Yes. MANYIKA: Well— LAGARDE: That’s my life, so—(laughter)— MANYIKA: We’re going to go to the— LAGARDE: Maybe other— MANYIKA: We’re going to go to the audience for some questions. And if you have a question, please put up your hand. And we’re on the record. Just say your name and your affiliation. And please ask a question. There’s one right there. Q: Good morning. I’m Lauren Leader. I run All In Together, which is a women’s civic-education organization here in the U.S. On the subject of civics and politics, I haven’t had a chance to read the Council’s certainly stellar report. I’m astonished often at how rarely the topic of women’s political leadership and participation comes up in the context of economic participation. The WEF looks at those things in consort. So the U.S., for instance, is 96th in the world on political participation, though number one in the world for educational attainment. As you mentioned, Tunisia, that’s what triggered it, because Tunisia has had some very progressive laws around women’s political representation in their parliament. Could you talk about the impact of women’s political representation in parliaments and the governments on their seriousness with which they look at closing the labor force participation, addressing some of these larger economic issues? I think they’re inseparable, but I continue to see them treated very separately in the sort of global dialogue about that. LAGARDE: Very good point. First of all, global and average numbers are abysmal, whether it’s in parliament, in government, heads of states. The number of women—I think the heads of states, we have eleven at the moment heads of government, twelve, and the average participation in parliament is around 20-ish percent. So that’s really not good. In countries where—second point. I think that quotas can actually make a difference. I’ve seen it and I’ve witnessed that in my country, where quotas were imposed at local level gradually, then at the senatorial levels, and when we had—and as a result of that, we simply had a very large and growing numbers—number of women in parliament; not there yet, but moving. Third point: In those countries where, either because of quotas or because of the political determination of people or because of the scarcity of resources, the numbers look much better. And obviously you will think at this point of the Nordic countries. Surprise, surprise—they have much higher participation of women in their respective organizations at executive and legislative levels, and they also have the smallest gender gap when it comes to female labor force participation, although no country has actually effectively closed that gap. I think the lowest gap ever in the world is seven. And no country has gone below that. But I also think of countries like Rwanda or like Ethiopia, where in Rwanda you have 61 percent female participation in parliament. In Ethiopia, I think the goal is 50 percent for the next election. And the government itself in Ethiopia is now 50 percent female. In Rwanda it’s a little less than that. But when I look at Rwanda, there are notable things that can be said about Rwanda, but you cannot deny that it’s efficient, that it’s well run, and that the development indexes—and maybe Kristalina will talk to that more than me—are quite stellar. One more, actually, which I’m taking politics into—hiring and retaining people and promoting them along the way has a political dimension. And in the field that I know well, when you look at the banking sector and you look at those banks and those financial institutions where you have a higher proportion of women than, again, the abysmal number of 20 percent board members or members of executive committees, and 2 percent of CEOs, when you look at those institutions where the percentage is much higher, you also realize that the nonperforming loans—surprise, surprise—it’s correlation, not causality. But correlations can speak volumes. Volume of nonperforming loans, much lower; risk index, much better; and buffers in terms of capital, much higher. Quote, for me, that means safer. MANYIKA: There’s a question in the back that went up first. Q: Hi. Good morning, and thank you so much. My name is Amina Tirana. I’m with Visa, Inc. And I’d like to thank you especially for bringing attention and calling out both the low-income countries but also the question of informality. And I’d love to hear some of your thoughts, maybe on a couple of policy priorities or recommendations, for addressing the informality of labor and women in many of these countries. And it’s something that at Visa we’re thinking quite a bit about, especially for micro- and small-business owners, who are people who have what we call livelihoods or even subsistence business in retail, where the goals—women are often tied to home. They can’t move for jobs. And the goals, as we do more and more research, not always about growing and becoming a big enterprise, but about security of income, stability for the family, and providing for the next generation. They don’t want their children to have to do what they’re doing as well. Often those businesses are informal or quasi-formal. Do you have recommendations about how we can help them thrive and meet their goals? LAGARDE: First of all, I think that we should be able much better to measure that and to really take that into account in any of those growth measurements and economies’ measurements, because what you can’t measure, you can’t actually influence very much. So we operate on the basis of assumed informality numbers. But we don’t know enough. We don’t have enough in that respect. As a follow-up to that, in a way, I think that technologies can actually help a big way. If you look at countries like India or Kenya, I mean, they’re the most typically quoted references, but there are other countries as well which are using, you know, digital payment, mobile applications, that can actually identify economic operators, and in many instances simplify their life, but also measure what transactions are conducted, what business is generated. Third point, I think you can design taxations and you can design your fiscal principles in order to not repress that activity but bring it into the fold by not assessing massive either social-security charges or tax charges in a brutal and heavy way, but do it very gradually and very incrementally in order to bring about that informality. Fourth thing, which I believe also matters a lot for women, is to put in place those cash-transfer systems that have been used in some instances with technologies in the process in order to eliminate the risk of theft violence against women that apply whenever there is real cash being transferred and carried around; and second, encourage other factors such as girls in education or less informality and better access to services. It’s all linked, and you really have to treat that in a very comprehensive way. But I think the means—I think we’re getting more tools to reach those objectives. MANYIKA: The person right here in front. Q: Thank you so much for being here this morning. My name is Nili Gilbert. I’m a co-founder and portfolio manager at Matarin Capital. My question for you is specifically about women in control of capital. As you know, when it comes to moving major institutional capital around the world, much of the money is being moved by men. And I think a lot, as a portfolio manager, about how the backgrounds of the people on our team making decisions about money affect the way they move the money, the decisions that they make. I wonder, you know, particularly for a woman like you in your role or Ms. Georgieva at the World Bank, whether you think that if there were more women making decisions about how the major pools of capital are moving around the world, would it have the potential to actually change the structure of the capital markets or the types of projects that capital is flowing to. Thank you. LAGARDE: That’s a really—I’m learning from you as much as I’m trying to think about the response. I’m known to have said that in 2008, if Lehman Brothers had been Lehman Sisters, we would not be—(laughter)—in the mess we were in and continue to be. (Applause.) So I don’t know what it would look like if there were more women involved in those portfolio managements and in those financial flows that we see around the world. I don’t know because I—we don’t have the counterfactual, because there are so few women in those businesses. So it’s hard to imagine. But if we extrapolate a little bit—and based on the work that we’ve done on banks and financial establishments, I would assume that the business conducted in those fields would be less risky, would be safer, would probably be better hedged, and possibly, when it comes to the financing of infrastructure projects, for instance, would take into account the impact on women’s life. So if—you know, I’m dreaming here, but if policymakers and the financiers of those decisions were more women, had more women, maybe the choice between a huge big arena—(laughter)—France World Cup, yes, but—but—(laughter)—and a major project that would help, you know, with water, you know, a good, sensible dam with irrigation projects associated with it. If you—if more women were involved in that choice, maybe—maybe—the choice would go in the direction of water than soccer. (Laughter.) MANYIKA: We’ll take a couple more. One over there? Q: Thank you very much. My name is Janet Fleischman with the CSIS Global Health Policy Center. Melinda Gates had said last year that no country has emerged from poverty without expanding access to contraceptives. And I wonder if you could speak about the enabling environment for economic empowerment for women and the importance of access to women’s health services, to family planning, and the impact that this has in countries around the world, including in the U.S., but certainly in many of the developing countries as well. LAGARDE: You know, where we have the most empirical evidence is in relation to low-income countries where you see—where you have seen over the course of time an empirical evidence of the fact that when education is longer, when family size are smaller, it’s an—well, sorry, I shouldn’t—I’ll work it around the other way. When you have reduced—it’s not reduced fertility, it’s reduced natality, it’s invariably caused by longer and higher levels of education for women. So the linkages between these two factors, education for women and number of children, is obvious. The same goes for age of marriage. And we’ve done a study on India in that respect where you can actually very visibly see that with an additional number of compulsory education, the age of marriage moves into time; and therefore, the number of children subsequently is slightly reduced. But that’s, I mean, that’s the best that we can produce in that respect and it’s obviously as a result that in order to develop, grow an economy, you need to provide the level of education that will actually maintain fertility at an acceptable rate. And I agree with Melinda. MANYIKA: Yeah. Actually, there was a terrific study that I think U.N. Women did that actually included that. LAGARDE: Yes. MANYIKA: It also even included factors like physical safety, for example, how the linkages— LAGARDE: Transportation— MANYIKA: Transportation, right. LAGARDE: —comes to mind. You know, access to proper sanitation is another one. Yeah. MANYIKA: Right. There’s a question right in front here. Q: Thank you. LAGARDE: I remember, by the way—this is—this is a funny memory that you bring back because you mentioned Melinda. But we had a fantastic conversation in India with the prime minister in attendance about the size of sanitation and the height of walls behind which people could do what they have to do and how it impacts on the freedom of movement of women and their ability to actually go to work or leave the house. We don’t think about it because we are in the U.S. and that’s not something that we’re exposed to, but boy. Sorry for that discretion. Q: No, it’s a terrific one. And thank you so much and such an inspiration to hear you and also Kristalina when we get to her conversations around these issues. I loved your piece, your response in terms of this is a—this is a journey, this isn’t something that’s going to turn around in a short period of time, and just kind of linking this to the various dimensions of this question of economic opportunity, including the political. But also, I’d love to hear your thoughts around the social context as well because we’ve been—we’ve seen evidence over time and we’ve been learning as we’ve done more research and studies. But at the heart of it, we’re talking about social constructs and values, I think, in communities in terms of how women are perceived in society. And I would love to hear your comments around where, if there are any gaps, or how could we look differently at those, how could we respond differently, I think, to those questions as we are putting forth the evidence that you’ve just described in terms of the gap numbers, the impact on women, the questions around whether women’s participation in some of these decision-making would actually make a difference, et cetera. But getting back to your thoughts just around that whole, at the end of the day, the social construct that we’re dealing with around how women are perceived and values that we have. LAGARDE: I’d be out of my depth to address your question. So you might be yourself better placed in a position to address those issues. (Laughter.) You know, I can only speak to what we try to measure, what we try to influence, and based on honest facts—honest facts—and numbers, yes—(laughter)—and my—and my own history. But, you know, I’m deeply convinced that it is a revolutionary process because it changes, it disrupts. It shouldn’t displace and it shouldn’t be conducted in a way that is aggressive and adversarial because I think everybody stands to benefit from that. I have other beliefs, but that’s irrelevant. (Laughter.) MANYIKA: We’ll take probably two more. One over there on this side. Q: Nan Cohen, Princeton. Thank you for being here. I wanted to go back to part of an answer you gave earlier about the ways in which including more women brings greater diversity and, therefore, greater productivity. Could you say something? Used the word “characteristics”—what sorts of characteristics, given that we’re all individuals and very varied, what sorts of characteristics do women in general bring that strengthens an organization? LAGARDE: The easy answer to that without taking sides—and I’ve indicated on which side I am, right—but I think the first easy answer is diversity in and of itself, because we look different, we have different backgrounds, we have different approaches to life in many instances, all of us. And it’s not just diversity based on gender, it’s a diversity that is based on gender, on colors, on religion, on background, on education. You know, from Princeton, you come with a particular background. If I come from another vocational school somewhere, I have a different background and we both equally can contribute, yet we will bring diverse contributions. The simple fact that there is diversity I think enriches the debate and forces you, forces me to listen to somebody who comes from a different background, different religion, different framework of mind. We were talking about that earlier on. You know, how do you put yourself in the shoes of somebody who’s been trained and educated in China or in the Middle East or in a civilization and with a cultural background that is so different from yours and from mine? That in and of itself because it leads you to doubting and questioning and arguing for your own sake. That is, I think, a plus and enriches and makes the contribution—the output as they call it—more productive. I also think for myself—and that’s only based on my life in, you know, twenty-five years of private sector, now soon ten years in public sector—I believe that men and women react in a different way to situations. It’s overstated as a generality. I would say that a large majority of the responses, I can, you know, blindly contribute to the playing of music, for instance, which— MANYIKA: Right. LAGARDE: —even behind a curtain, yeah, I think that there are differences in terms of being more prone to taking risks or not, thinking ahead, which is why I was thinking that maybe that world of portfolio management would be less risky and better hedged. I’m sorry to say, but thinking of others rather than thinking about oneself also comes to mind. And I don’t know whether it’s, you know, determined by nature, whether it’s determined by history, whether it’s determined by education, I don’t want to opine on that. But I have seen for myself those clear distinctions and we are seeing it in portfolio management as well and in—and in banks. Again, big, strong correlations do not justify causality necessarily, but it’s pretty strong. MANYIKA: Over to this side. Thank you. Q: Hi. Thank you so much. My name is Nancy Lieberman. I’ve been a partner at a major international law firm, Skadden, Arps— LAGARDE: Yeah. Q: —for decades. I’ve been there nearly forty years. And I do M&A law. And my experience in addressing corporate boards has led me to conclude that if there were more women directors, just as you were answering the last question, it would make a sea change difference in so many areas that impact equality of women in the workplace. I know that the EC now has a quota system for demanding that women be on corporate boards. I personally am not a great believer in quotas because I think it diminishes what a woman brings to the party if the label is “I’m a quota person.” So I don’t like that. So my question to you is—and I have served on a corporate major board, so I’m familiar with the process—what do you think will change the way boards choose directors to get more women on? Because from my own personal experience, I think there’s a huge amount of resistance still. LAGARDE: Yeah. Q: So what’s your solution to the problem? LAGARDE: I’ll give you a very practical example. And by the way, I’m almost on the same page as you and I was totally on the same page as you until I became chairman at Baker & McKenzie. And I looked at the number of female partners and I looked at the population we had and I looked at the gap and I thought solve it, we need quotas, and I’ve completely changed my mind, not forever, but the step is just too high and we need—we need—we really need it. And I think what we’re seeing in some of the—some of the European countries—and France was, you know, on the—on the forefront of requiring that there be, first of all, 30 percent and then soon 40 percent of women on boards. I’ll give you my personal experience of that because I think that it goes down to the practical details and the weeds of what we do. I was finance minister for a period of four years right at the time of the financial crisis, so a tough job to do. But I had a portfolio myself of state-owned or partly state-owned companies and not, you know, small things. You talked about, you know, the big telecom companies, the big utility companies, you talk about airports, you talk about airline companies, all those. So it didn’t exist at the time, but I said I would like to meet each and every one of the chairmen or CEOs. It’s good enough to be represented at the board by some of my civil servants—fine—but I want to see them, and I’m going to invite them into my office to hear about their strategy because I just want to hear. So they all came and, invariably, I finished those strategy-dedicated meetings to, of course, you have women on your board? And invariably, the answer was, uh, no, or sometimes, oh, yes, of course, I have one. (Laughter.) Q: It’s still like that, by the way. LAGARDE: Huh? Q: It’s still like that. LAGARDE: I know. Yeah. And so we all have to continue doing that. So I said, well, why is that? And they said, well, you know, we’re making an effort by blah, blah, blah, blah, blah, blah, it’s very difficult to find. So I said, well, you know what? When we see each other next year, I hope you’ve made progress and you’ve identified board members that will have joined your board because you’ve got a couple of very aging gentlemen here—(laughter)—that would be better off doing something else. So comes the next year. I knew what I was going to get, I just knew it, so I was prepared for that. So they all came, and I got invariably, except for one, the same answer: Minister, I tried so hard, but I couldn’t find a competent woman to do the job. So I said, oh, very good. You know what? And I pulled out of my pocket a list of about thirty women, who were not only competent, but willing and just ready to serve. So I said next year, you just come and see me, and if you haven’t changed your board composition, you’re in real trouble. And that changed. And it’s at that time that the parliament actually passed the—I did not cause that law, you know, in fairness, I supported it, but that’s when they passed the law on minimum threshold in all boards of publicly quoted companies. And now it’s moving to the medium-size companies as well where there’s a huge, big, big gap. MANYIKA: We’ll take our last question over there. Q: I know how you feel when you go to a meeting where you are singled out somewhere. (Laughter.) But I am a philanthropist and I work with the United Nations, with the IMF, and the World Bank, with UNESCO. And I want to share a personal story about my idea about the women’s empowerment. I had a chance to serve with Hillary Clinton on a project in Amman, Jordan for a museum fund for international artists. And the UNESCO was looking for a philanthropist, you know, and they were not able to find someone who was committed to women’s empowerment, who liked the Muslim culture, and who had the incentive to go into such a project, but I was the one. So you (just find ?) about my sorrow when Hillary lost her election in November 2015 (sic; 2016). I tried to recover from that. But my point is the fact that, if I am a women-empowerment-committed man, the reason is because when I was six years old in the kitchen, my mother explained to me the fact of life, that women cannot drive, not in the city, cannot borrow money, cannot own money, cannot have property, and everything. I said, wow, this is incredible, I want to fight against that. So my suggestion—and by the way, there is something good that is happening right now. Three weeks ago I was at Goldman Sachs for a (third year ?). Normally, there is one woman there and then, yes, there’s one there, but this year there was a lot of women. And I said, my goodness, something happened here. And in the panel, the same thing. There was a new CEO at Goldman Sachs and then he told us this is something I want to achieve. But my suggestion, why not train, implement, influence young kids at six years old in the elementary school, even preschool, about what is a woman, why she’s good, why you should look at her and change your mentality early on. This will change a lot of things. And then probably two, three, five years ago, I will have (half ?) women interested by this subject because it is very, very crucial. Thank you. LAGARDE: Thank you so much. You know, I don’t—I don’t think anybody can disagree with you. I would go one step further. I believe—I strongly believe that everywhere we go, everywhere we are, we can actually have that at heart and make sure that we identify in our family life, in our professional life, in our day-to-day life how we can better respect, better access, and better encourage. Because, you know, when I started at the IMF, macroeconomists in the main said, oh. So even there, now there is a very strong current of people who say, yes, of course it matters, yes, it is macro-critical. So when you’re a six year old, yes, absolutely, because you determine, and even before that possibly, and on and on and on and wherever and forming teams and foraging alliances with both men and women. Yes. MANYIKA: Well, that’s a wonderful note to end on. I just want to thank you for taking the time, Christine, to have this conversation, to come here on this macro-critical issue. (Laughter.) Thank you. LAGARDE: You got it. (Applause.) (END)

omics journal

Medicinal Plants Used In The Ejisu-Juaben Municipality, Southern ... https://www.omicsonline.org/.../medicinal-plants-used-in-the-ejisujuaben-municipality... Medicinal plants used in the Ejisu-Juaben municipality, Southern Ghana: An ... Aim: The aim of this study was to explore and document the use of medicinal

Mean feet: the tap-dancing duo who were Fred Astaire's heroes

https://www.theguardian.com/stage/2016/oct/06/tap-dancing-fred-astaire-fabulous-nicholas-brothers Blending a floating grace with a fierce athleticism, the Fabulous Nicholas Brothers won fans from Gene Kelly to Michael Jackson. Now a cinematic retrospective celebrates their talents Judith Mackrell @judithmackrell Thu 6 Oct 2016 07.00 BST Last modified on Mon 10 Oct 2016 16.42 BST Astonishing … the Nicholas Brothers – Fayard, left, and Harold – in 1943’s Stormy Weather. Photographs: Twentieth Century Fox Film Corporation/Photofest When connoisseurs of dance are asked to compile a list of the world’s greatest movers, Fred Astaire is nearly always close to the top. As a dancer, he brought a ballroom finesse and a loose-knit debonair grace to the grounded rhythms of tap. As a choreographer, he was remarkable for the puckish playfulness of his imagination: playing the drums while he danced in the 1937 film A Damsel in Distress; dancing with his shadow in Swing Time (1936); and revelling in the possibilities of trick photography as he danced up the walls and across the ceiling in Royal Wedding (1951). Yet Astaire himself declared that his own tap heroes were Fayard and Harold Nicholas, two black dancers who became one of the most popular double acts of the mid 20th century. During the 1930s and 40s, the brothers toured the world and acquired an international public through Hollywood movies such as Down Argentine Way and Stormy Weather. Nor was it just Astaire who rated them professionally – over the years, dance luminaries such as Gene Kelly, George Balanchine, Mikhail Baryshnikov and Michael Jackson added their own accolades. Tap devotees still prize the Fabulous Nicholas Brothers, but to the general public their names have become less well known. Picturehouse Cinemas’ retrospective of their work should lead to a deserved reappraisal of their importance. Fayard (born in 1914) and Harold (born in 1921) grew up in Philadelphia, where their parents led the resident band of the Standard theatre. It was the heyday of black vaudeville in the States – a period when great African American performers such as Bessie Smith toured the country. And much like Josephine Baker, who learned to dance by imitating the acts she’d seen at her local theatre in St Louis, the brothers acquired their skills at the Philadelphia Standard, as Fayard studied acts including Willie Bryant and Bill Robinson, then passed his skills on to his younger brother. Stormy Weather (1943) Directed by Andrew L. Stone Shown from left: Fayard Nicholas, Harold Nicholas Fabulous Nicholas Brothers press image supplied by Gemma Cole Production Manager elevenfiftyfive.com 07794166846 Facebook Twitter Pinterest Natural mimics as well as naturally talented, the Nicholas boys also picked up tricks from the acrobats and comedians who appeared on the Standard stage. By the time they started performing themselves, around 1930, they had developed their own unique physical virtuosity, and by 1932 they’d moved up to New York, starring at the Lafayette vaudeville theatre in Harlem and at the Mecca of black dance and music, the Cotton Club. Sign up to our Film Today email Read more Given the brutal segregation that operated within the entertainment industry, it was hard for black entertainers to break into the white mainstream. But the Nicholas brothers were among the privileged few, like Baker and Smith, whose talent was too irresistible a commodity. In 1936, when Harold was still virtually a child, they were hired to perform in the Ziegfeld Follies on Broadway, where their dancing created such an uproar that the singer and comedienne Fannie Brice, who came on stage after the brothers, had to quiet down the theatre by cracking the line: “Do you think we can talk now?” The following year, ballet choreographer George Balanchine hired the brothers to dance in the musical Babes in Arms. Hollywood, too, came calling, and during the 1940s the two men featured in some dozen major films, their bravura skills showcased in sequences such as this Chattanooga Choo Choo routine, in the 1941 movie Sun Valley Serenade. The brothers did not make the transition on screen from dancers to romantic leads, as Fred Astaire had done the previous decade. Their skin colour was an obvious barrier, although Astaire himself would think nothing of blacking up his face to perform the “Mr Bojangles” number in Swing Time. But as dance artists the brothers pushed themselves constantly, absorbing new ideas and taking influences from ballet, a form they greatly admired. In performance they seemed both fearless and joyous. Baryshnikov declared they were the most “amazing dancers” he’d ever seen; and their most astonishing screen performance, the finale to the 1943 film Stormy Weather, was adjudged by Astaire to be the “greatest dance and music” sequence in the history of cinema. Facebook Twitter Pinterest Watching this clip, its striking how similar the brothers were to Astaire in certain ways; although physically chunkier, they brought the same quality of upper-body float to their dancing, the same easiness in their carriage and arms. It’s a lightness that informs their glissando slides (at 2mins 45secs) and the insouciant timing of their jumps – even when they’re leaping hazardously from table to table, over the band (0.32) or up a flight of stairs, the brothers work that special dance miracle of stretching out the seconds, of catching an impossible breath mid-air. Like Astaire their phrasing is impeccable, not least in the witty call-and-response section they perform with their pianist (1.30). Almost any other dance act would have milked this moment for much longer – Astaire certainly would have done so – yet here its only a glorious joke, a brilliant bagatelle before the brothers move on to other feats. The special hallmark of their act was its athleticism, however: the brothers’ ability to incorporate spins, kicks and flips into the middle of riotously fast and jazzy footwork; their special stunt of sliding down into the splits and back up again (1.36), as if friction and gravity were meaningless. The climax of this Stormy Weather clip almost beggars belief, as the two men make a leapfrogging descent of the staircase, taking it in turns to jump over each others heads, each time landing in the splits and rebounding back up. All this they manage in full evening dress, and without a moment’s hesitation flickering through the mega-wattage of their exuberance and charm. Gregory Hines, no mean dancer himself, said of the Nicholas brothers that it would be impossible to make a movie about their lives because it would be impossible to cast. No two dancers could ever be found to match their a skills, Hines believed, and the revelations of this three-minute clip suggest he may have been right. The Fabulous Nicholas Brothers season is at selected cinemas from 19 October–4 November as part of the BFI’s UK-wide season Black Star.

Herbal fertility treatments used in North America... now on PubMed

Elsevier Dear Dr. Lans, The files used to generate your final article, Herbal fertility treatments used in North America from colonial times to 1900, and their potential for improving the success rate of assisted reproductive technology in Reproductive Biomedicine & Society Online, have been published on PubMed Central (PMC). http://www.ncbi.nlm.nih.gov/pmc/articles/6047296

Hazelnuts 365: Why Oregon's state nut may be the key to disease prevention

Public Release: 11-Dec-2018 https://www.eurekalert.org/pub_releases/2018-12/fh-h3w121018.php New study finds daily consumption of hazelnuts improves biomarker of vitamin E status in older adults Oregon Hazelnut Marketing Board IMAGE IMAGE: 99% of U.S. hazelnuts are grown in Oregon view more Credit: Oregon Hazelnut Marketing Board Dec 10, 2018 - Aurora, Ore. - Hazelnuts are poised to be the "it" nut for 2019, and new research suggests that adding hazelnuts to your daily diet could bode well for long-term health. The new study, administered by the Linus Pauling Institute at Oregon State University and published in the December 2018 issue of The Journal of Nutrition , found that older adults who added hazelnuts to their diet for 16 weeks significantly improved their levels of two key micronutrients. Results showed increased blood concentrations of magnesium and elevated urinary levels of a breakdown product of alpha tocopherol, commonly known as vitamin E. Older adults are at increased risk of various chronic diseases where inadequate levels of vitamins and minerals may play a significant role, including cardiovascular disease, Alzheimer's disease, liver disease and cancer. Tree nuts, including hazelnuts, contain a wide variety of vitamins and minerals, and are an excellent source of vitamin E and good source of magnesium, two "shortfall nutrients" that are lacking in the typical American diet. Study Details Researchers stated the objective of the study was to determine whether daily hazelnut consumption by healthy older adults for 16 weeks improves biomarkers of micronutrient status, especially vitamin E and magnesium. Participants (n = 32 including 22 women; mean ± SD age: 63 ± 6 y) consumed hazelnuts (?57 g/d) for 16 weeks. Blood and urine samples and anthropomorphic measures were taken at the start and end of the intervention to determine plasma concentrations of α-tocopherol and serum concentrations of magnesium, lipids, glucose, insulin, and high-sensitivity C-reactive protein along with urinary vitamin E metabolites; several other micronutrients were measured by a lymphocyte proliferation assay. There were 3 primary endpoints, calculated as the mean changes in measurements between baseline and the end of the 16 week intervention for 1) plasma α-tocopherol, 2) urinary α-carboxyethyl hydroxychromanol (α-CEHC; an α-tocopherol metabolite), and 3) serum magnesium. Hazelnuts: The indulgent health nut A one-ounce serving (28.35 g) of raw hazelnuts contains 27 percent (4 mg) of your daily value (15 mg) of vitamin E . Vitamin E is a shortfall micronutrient, as identified by the Dietary Guidelines for Americans 2015-2020 , which frequently is consumed at levels less than the Estimated Average Requirement of 15 mg/day. These new findings complement existing knowledge about the role of nuts in heart health. In 2003, the U.S. Food and Drug Administration approved a qualified health claim related to nuts that states: "Scientific evidence suggests but does not prove that eating 1.5 ounces per day of most nuts, such as hazelnuts, as part of a diet low in saturated fat and cholesterol may reduce the risk of heart disease." U.S Dietary Guidelines recommend that the majority of your fat intake be unsaturated. One serving of raw hazelnuts (28.35 grams, or about 21 hazelnuts) has 6 grams of monounsaturated fat and only 1 gram of saturated fat. This discovery also builds upon a growing body of scientific evidence on the benefits of nuts for older adults (average age 63 + 6 years). In 2013, observational researchers at Harvard University looked at how eating nuts may help reduce the risk of mortality, finding that those who ate nuts daily, such as hazelnuts, saw health benefits nearly double. The benefits were seen in both men and women, independent of other predictors for mortality. The results, published in the New England Journal of Medicine , are based on approximately three decades of follow-up among 76,464 women in the Nurses' Health Study (1980-2010) and 42,498 men in the Health Professionals Follow-up Study (1986-2010), including examination of food questionnaire data. Compared to people who didn't eat nuts, people who ate nuts saw benefits that increased along with the number of servings of nuts they ate. Those who ate nuts seven times a week had nearly twice the benefit compared to those who ate nuts once a week; once a week nut eaters had a small, but still significant benefit. This observational study is an important addition to the body of research on nuts and heart health; however, given its observational nature, it's not possible to conclude cause and effect between nut consumption and mortality. Hazelnuts 365: A tasty way to get your daily dose of nuts Oregon hazelnuts are a delicious way to get seven servings of nuts per week. Because they are so versatile, these crunchy Pacific Northwest gems upgrade the flavor and nutrition of salads, entrees, desserts and snacks. A one-ounce serving of hazelnuts equals approximately 21 nuts. A 2017 consumer survey, funded by the Oregon Hazelnut Marketing Board, found that 47 percent of people found hazelnuts to be "very healthy," which was nearly twice the number from the previous year. The survey also found people don't view hazelnuts as being as expensive as some other nuts. Food manufacturers have taken note and hazelnuts are gradually starting to appear in more commercially available products, according to the survey, growing from 63 products in 2013 to 93 in 2015, when data was last available. Oregon boasts an ideal climate for producing the world's highest quality hazelnuts and it's in this special corner of the world where temperate ocean, mountain and river climates meet with rich volcanic soils to create prime hazelnut-growing country. Ninety-nine percent of U.S. hazelnuts are grown in Oregon across 72,000 acres. The 2018 harvest has officially come to an end, and early reports indicate a yield of 46,000 to 48,000 tons, an increase of 44-50 percent over last year's 32,000 tons. "This harvest season was a great success thanks to perfect weather conditions, new acreage, dedicated farmers and the hazelnut industry work force," said Meredith Nagely, manager of the Oregon Hazelnut Marketing Board. "Consumers can expect to see an increase in hazelnuts available at retailers and on menus." ### The Oregon State University Foundation and the Oregon Hazelnut Marketing Board funded this research. About Oregon Hazelnut Marketing Board The Oregon Hazelnut Marketing Board was established in 1949 by the growers and handlers of hazelnuts. The purpose of the board is to set quality standards for the industry, ensure all imported product meet U.S. standards and provide funding for promotion of hazelnuts through research, education and promotion programs. For more information, visit oregonhazelnuts.org.

Green leafy vegetables may prevent liver steatosis

https://ki.se/en/news/green-leafy-vegetables-may-prevent-liver-steatosis Published 2018-12-17 21:00. Updated 2018-12-18 19:05Denna sida på svenska Nitratrik salladsblandning. Foto: iStock A larger portion of green leafy vegetables in the diet may reduce the risk of developing liver steatosis, or fatty liver. In a study published in PNAS researchers from Karolinska Institutet in Sweden show how a larger intake of inorganic nitrate, which occurs naturally in many types of vegetable, reduces accumulation of fat in the liver. There is currently no approved treatment for the disease, which can deteriorate into life-threatening conditions such as cirrhosis and liver cancer. Liver steatosis, or fatty liver, is a common liver disease that affects approximately 25 per cent of the population. The most important causes are overweight or high alcohol consumption and there is currently no medical treatment for the disease. Researchers at Karolinska Institutet have now shown how a greater intake of inorganic nitrate can prevent the accumulation of fat in the liver. “When we supplemented with dietary nitrate to mice fed with a high-fat and sugar Western diet, we noticed a significantly lower proportion of fat in the liver,” says Mattias Carlström, Associate Professor at the Department of Physiology and Pharmacology, Karolinska Institutet. Their results were confirmed by using two different cell culture studies in human liver cells. Apart from a lower risk of steatosis, the researchers also observed reduction of blood pressure and improved insulin/glucose homeostasis in mice with type 2 diabetes. The research group’s focus is the prevention of cardiovascular diseases and type 2 diabetes through dietary changes and by other means. Previous studies have shown that dietary nitrate from vegetables enhances the efficiency of the mitochondria, the cell’s power-plant, which can improve physical endurance. It has also been shown that a higher intake of fruit and vegetables has a beneficial effect on cardiovascular function and on diabetes. Mattias Carlström. Foto: Privat “We think that these diseases are connected by similar mechanisms, where oxidative stress causes compromised nitric oxide signalling, which has a detrimental impact on cardiometabolic functions,” says Dr Carlström. “We now demonstrate an alternative way to produce nitric oxide, where more nitrate in our diet can be converted to nitric oxide and other bioactive nitrogen species in our body.” Even though many clinical studies have been done, there is still considerable debate about what properties of vegetable make them healthy. “No one has yet focused on nitrate, which we think is the key,” continues Dr Carlström. “We now want to conduct clinical studies to investigate the therapeutic value of nitrate supplementation to reduce the risk of liver steatosis. The results could lead to the development of new pharmacological and nutritional approaches.” While larger clinical studies are needed to confirm the role of nitrate, the researchers can still advise on eating more green leafy vegetables, such as regular lettuce or the more nitrate-rich spinach and rocket. “And it doesn’t take huge amounts to obtain the protective effects we have observed – only about 200 grams per day,” says Dr Carlström.“ Unfortunately, however, many people choose not to eat enough vegetables these days.” The study was financed by the Swedish Research Council, the Swedish Heart and Lung Foundation, Novo Nordisk, the European Research Council and Karolinska Institutet. Two of the authors, Jon O Lundberg and Eddie Weitzberg, are coinventors on patent applications related to the therapeutic use of inorganic nitrate. Magnus Ingelman-Sundberg is cofounder of the contract research organisation (CRO) HepaPredict AB. Publication “AMP-activated protein kinase activation and NADPH oxidase inhibition by inorganic nitrate and nitrite prevents liver steatosis”. Isabel Cordero-Herrera, Mikael Kozyra, Zhengbing Zhuge, Sarah McCann Haworth, Chiara Moretti, Maria Peleli, Mayara Caldeira-Diaz, Arghavan Jahandideh, Han Huirong, Josiane Cruz, Andrei Kleschyov, Marcelo Montenegro, Magnus Ingelman-Sundberg, Eddie Weitzberg, Jon O Lundberg and Mattias Carlström. PNAS, online 17 December 2018, doi: 10.1073/pnas.1809406115. 18 Dec 2018 by Kommunikationsavdelningen.

Aging-related premature luteinization of granulosa cells is avoided by early oocyte retrieval

in Journal of Endocrinology Authors: Yan-Guang Wu 1 , David H Barad 1 , 1 , 1 , Vitaly A Kushnir 1 , 1 , Emanuela Lazzaroni 1 , Qi Wang 1 , David F Albertini 1 , 1 and Norbert Gleicher 1 , 1 , 1 View More DOI: https://doi.org/10.1530/JOE-15-0246 Page(s): 167–180 Volume/Issue: Volume 226: Issue 3 Article Type: Research Article Online Publication Date: Sep 2015 Copyright: © 2015 Society for Endocrinology 2015 Free access Citation Alert Citation Alerts Get Permissions Download PDF Check for updates Abstract/Excerpt Full Text PDF Abstract Why IVF pregnancy rates decline sharply after age 43 is unknown. In this study, we compared granulosa cell (GC) function in young oocyte donors (n=31, ages 21–29), middle-aged (n=64, ages 30–37) and older infertile patients (n=41, ages 43–47). Gene expressions related to gonadotropin activity, steroidogenesis, apoptosis and luteinization were examined by real-time PCR and western blot in GCs collected from follicular fluid. FSH receptor (FSHR), aromatase (CYP19A1) and 17β-hydroxysteroid dehydrogenase (HSD17B) expression were found down regulated with advancing age, while LH receptor (LHCGR), P450scc (CYP11A1) and progesterone receptor (PGR) were up regulated. Upon in vitro culture, GCs were found to exhibit lower proliferation and increased apoptosis with aging. While FSH supplementation stimulated GCs growth and prevented luteinization in vitro. These observations demonstrate age-related functional declines in GCs, consistent with premature luteinization. To avoid premature luteinization in women above age 43, we advanced oocyte retrieval by administering human chorionic gonadotropin at maximal leading follicle size of 16 mm (routine 19–21 mm). Compared to normal cycles in women of similar age, earlier retrieved patients demonstrated only a marginal increase in oocyte prematurity, yet exhibited improved embryo numbers as well as quality and respectable clinical pregnancy rates. Premature follicular luteinization appears to contribute to rapidly declining IVF pregnancy chances after age 43, and can be avoided by earlier oocyte retrieval. Introduction Effects of female reproductive aging on assisted reproductive technologies are widely acknowledged. Age-related gradual declines in implantation and pregnancy rates (van Noord-Zaadstra et al. 1991) as well as increases in spontaneous miscarriages (Belloc et al. 2008, Grande et al. 2012) were observed. Declines in oocyte quantity and quality are considered principal driving forces through as yet ill-defined mechanisms (Navot et al. 1991). Based on annual reports to the Centers for Disease Control and Prevention (CDC), as required under a federal statute from IVF centers in the United States, our center is distinguished from most in serving the oldest IVF patient population. CDC reports further establish that with respect to the oldest age group of patients seeking infertility treatments (>43 years), for which almost no national data are available, our center is unique in accommodating a disproportionate number of patients of advanced age. While our center's IVF outcomes are reflective of the gradual decline in pregnancy rates normally observed with advancing female age, we have noted that this decline after age 43 sharply increases. Mechanisms leading to accelerated loss of ovarian function are unknown undoubtedly, however, related to the poor reproductive performance in women above age 43. Accordingly, we hypothesized that fast decreasing IVF pregnancy rates in older patients reflect poor ovarian environments, which adversely impacts oocyte/embryo quality and IVF outcomes (5). Specifically, we predicted that changes in immediate ovaria environments of oocytes, represented by granulosa cells (GCs), should be identifiable by comparing cell function at different maternal ages. Rapid declines in IVF pregnancy rates after age 43 may, therefore, reflect age-related functional decline in ovarian cells. These changes in the ovarian environment should be discoverable by comparing GC function. Oocytes in primordial follicles remain arrested in meiotic prophase I until recruited for oogenesis. Oocytes and accompanying GCs engage and maintain a symbiotic relationship (Buccione et al. 1990). GCs form the follicular microenvironment, which facilitates oocyte development, supplies energy, disposes of waste and participates in molecular signaling. If GC function becomes impaired with advancing age, oocyte growth and competence will be compromised in parallel. For example, GCs synthesize and transport energy substrates, nucleotides and amino acids into oocytes (Buccione et al. 1990). Using an in vitro oocyte growth model, Schultz et al. demonstrated that oocyte growth positively correlated with the number of adherent cumulus cells (CCs, representing more differentiated GCs) and the extent of metabolic cooperation between them (Brower & Schultz 1982, Herlands & Schultz 1984). Transcription in oocytes depends on the presence of attached CCs (De La Fuente & Eppig 2001). In addition, the maintenance of oocyte arrest before recruitment also relies on the contribution of autocrine and paracrine factors synthesized in GCs, including cAMP/cGMP (Webb et al. 2002, Wigglesworth et al. 2013, Shuhaibar et al. 2015), purine (Downs 1993), kit ligand (Ye et al. 2009), and NPR2 (Zhang et al. 2010, Tsuji et al. 2012, Wigglesworth et al. 2013). Oocytes from antral follicles resume and complete meiosis spontaneously after removal of surrounding CCs (Buccione et al. 1990, Mehlmann 2005), suggesting that CCs control oocyte nuclear maturation. Cytoplasmic maturation of oocytes also depends on CCs and GCs. Cumulus-oocyte complexes can mature and support embryo development after in vitro maturation (IVM). Removal of CCs before culture, resulting in denuded oocytes (DOs), leads to impaired oocyte and embryo development (Buccione et al. 1990). One striking example of metabolic cooperation between oocytes and surrounding CCs pertains to glutathione synthesis. Because glutathione derived from GCs and CCs is required for sperm decondensation and male pronucleus formation, lack of ability to produce glutathione in DOs restrains their development (Perreault et al. 1988, Zhou et al. 2008, 2010). Hence, when DOs are co-cultured with GC monolayer during IVM, glutathione levels are restored and developmental competence is reestablished (Zhou et al. 2008, 2010). Normal growth and maturation of the oocyte is thus a direct reflection of physiological status of the GCs. Certain defects of gene expression result in loss of GC function, which in turn can lead to reproductive dysfunctions. For example, follicle stimulating hormone receptor (Fshr) knockout in mouse GCs results in infertility due to lack of antral follicles (Dierich et al. 1998). Similarly, knockout of aromatase (Cyp19a1) (Fisher et al. 1998), IGF1 (Baker et al. 1996), estrogen receptor β (Esr2) (Couse et al. 2005) and androgen receptor (Ar) (Sen & Hammes 2010) in GCs leads to premature ovarian aging (POA) and female subfertility/infertility. To further prove the importance of GCs, Seifer and Sadraie reported significantly higher percentages of apoptotic GCs in infertile women, diagnosed with low functional ovarian reserve (Seifer et al. 1996, Sadraie et al. 2000). Other investigators reported diminished proliferation (Seifer et al. 1993), and high levels of mitochondrial DNA deletions (Seifer et al. 2002) in GCs of aged IVF patients. All of these abnormalities in GCs have the potential of contributing to decreased reproductive success in older women. Our study, therefore, reports on functional attributes of GCs derived from three groups of women: young oocyte donors (Group 1), middle-aged infertile women (Group 2) and old infertile women above age 43 (Group 3). As we demonstrate, in Group 3, a significant decline in GC function is detected that exhibits characteristics of premature luteinization. Based on these findings, we also report on results of a clinical pilot study of early oocyte retrieval in Group 3, which appear to ameliorate the negative impact of premature luteinization. This discovery provides insights into ovarian aging, and offers alternative strategies for improving pregnancy chances in older women. Materials and methods Patient populations and institutional review board The Institutional Review Board of the Center for Human Reproduction (CHR) approved this study in expedited review. Based on age, three distinct age groups were investigated (Table 1): oocyte donors represented the youngest (Group 1; n=31). By definition, they are young and carefully selected, meeting age-specific ovarian reserve parameters. Group 2 represented young to middle-aged infertility patients within an age range of 28–38 years (n=64) while Group 3 included the oldest infertility patients at 43–47 years (n=41). Table 1 Patient populations and cycle characteristics Group 1 Donors (n=31) Group 2 Intermediate age infertility patients (n=64) Group 3 Older infertility patients (n=41) Average age (years) 24.4±0.52a 34.1±0.38b 44.3±0.23c FSH (mIU/ml) 6.3±0.23a 7.6±0.57a 10.3±0.34b AMH (ng/ml) 3.1±0.23a 2.8±0.26a 0.28±0.08b Number of follicles/cycle 22.5±8.3a 10.5±7.1b 6.8±5.1c Number of oocytes retrieved/cycle 20.6±1.2a 9.8±2.5b 5.2±1.3c Number of MII oocytes retrieved/cycle 15.5±5.2a 7.1±2.3b 3.6±0.8c Number of artretic oocytes retrieved/cycle 1.3±0.4a 0.9±0.2ab 0.6±0.1b Number of embryo ≥4 cells 15.5±4.6a 7.1±0.89b 3.6±20c Pregnant rate/cycle 16 (51.6%)a 22 (34.4%)b 3 (7.3%)c Progesterone/estradiol ratio 0.26±0.08a 0.5±0.15b 1.96±0.47c Values with same letters in their superscripts in same row were not different significantly (P>0.05). n, number of patients. Patient and IVF cycle characteristics are shown in Table 1. All subjects underwent controlled ovarian hyperstimulation and oocyte maturation by human chorionic gonadotropin (hCG) according to previously described protocols (Gleicher & Barad 2011, Gleicher et al. 2013), followed by transvaginal ultrasound-guided oocyte retrieval. hCG was administered when leading follicles reached 19–21 mm. Oocyte donors were stimulated in a long gonadotropin releasing hormone agonist cycle (GnRHa, Lupron, leuprolide acetate, Takeda Pharmaceutical USA, Inc., Deerfield, IL, USA) with daily dosages of 150–300 IU of human menopausal gonadotropin (hMG) from various manufacturers. In contrast, infertility patients were stimulated in microdose agonist cycles (Lupron) with daily dosages of 450–600 IU of gonadotropins, typically in a majority (300–450 IU) administered as FSH, and in a minority (150 IU) as hMG. Follicular fluid was collected at time of oocyte retrieval only from follicles 15 mm or larger. Oocyte/embryo assessment and fertilization All media and reagents for IVF were purchased from LifeGlobal (Guilford, CT, USA). Oocytes collected at retrievals were cultured in HTF medium containing 10% human serum albumin (HSA) for 2 h before insemination. After removal of cumulus by hyaluronidase treatment, oocytes were assessed according to morphology. Oocytes with obvious first polar body (1st Pb) were identified as mature (MII); oocytes without 1st Pb were identified as immature (MI & GV); oocytes with brown dark color, cytoplasmic fragments and/or broken membranes were identified as atretic. Only MII oocytes were used for insemination. Fertilized embryos were cultured in vitro in Blastocyst medium (LifeGlobal) for 3 days, then were assessed according to their morphology. Embryos with 4–12 blastomeres of equal size and minimal cytoplasmic fragmentation were identified as good embryos, and designated as suitable for transfer or cryopreservation. Hormone measurement All serum hormone concentrations of patients were examined with AIA 900 Automated Immunoassay Analyzer (Tosoh, Minato, Japan) by following the instruction of user manual except AMH. Basic concentrations of FSH and AMH were determined on day 2 or day 3 of menstrual cycle. Progesterone (P4) and estradiol (E2) were measured on the hCG administration day. Regarding the user manual and installment instruction from the technique supports, coefficient of variation (CV) was performed by running one sample 20 times, then was calculated using the following equation: CV=(s.d.) (100)/mean. The results were analyzed by Tosoh technique support and listed as follows: CV (FSH)=1.1%; CV (E2)=2.1; CV (P4)=2.2. All CV values were verified as normal by Tosoh. The P4/E2 ratio was calculated as P4 (in ng/ml)×1000/E2 (in pg/ml). Serum AMH was measured commercially (LabCorp., Ramsey, NJ, USA). GC isolation Following retrieval, clumps of GCs were removed from follicular fluid. To avoid blood contamination, collected GCs were washed twice in D-PBS (Zenith Biotech, Guilford, CT, USA) by centrifugation (326 g, 5 min), and following PBS removal GCs pellets were either frozen at −80 ° for future use or prepared for in vitro culture. RNA extraction and real-time PCR Total RNA was extracted using TRIzol-reagent (Invitrogen) and 1 μg of RNA was reverse transcribed using RT enzyme (Invitrogen). cDNA amplification and quantification of PCR products was done with the StepOne real-time PCR system (Applied Biosystems) according to the manufacturer's instructions using Sybr Green (Invitrogen). Standard PCR settings (95 °C for 10 min, and 40 cycles of 95 °C for 15 s and 60 °C for 1 min, then dissociation stage for 15 s at 95 °C, 1 min at 60 °C, 15 s at 95 °C, and 15 s at 60 °C) were used. PCR primers and product information of all tested genes are listed in Supplementary Table 1, see section on supplementary data given at the end of this article. To avoid DNA contamination in PCR, primers pair must be separated by at least one intron (at least 5000 bp) and the corresponding genomic DNA. The specific PCR amplifications were validated by running melting curve analysis and gel analysis. The primers that have only one PCR product with correct size were chosen for the study. The efficiency of amplification was determined by running standard curve (efficiency of assay: 90–105%; R2>0.98; all Cq values were similar). Each sample was run in duplicate. For each target gene, the number of mRNA molecules was calculated and expressed relative to ribosomal protein L19 (RPL19) reference mRNA. To compare and calculate results from different PCR running, the normalization was performed as follows: each PCR run included a reference cDNA sample as control, which was made by mixing ten different patients' GCs. The results from different PCR runs were calculated according to these reference samples and final average gene expressions were then analyzed statistically. Western blot analysis All antibodies were purchased from Santa Cruz. GCs were homogenized in Ripa buffer (Sigma), and protein was purified as described by instructions. Protein concentrations were determined by using the Pierce BCA protein kit (Thermo Fisher Scientific, Rockford, IL, USA). Gel runs separated 20 μg of total protein and Ripa buffer was used as negative control. After electric transfer, membranes were blocked for 2 h with 5% nonfat dry milk. Then membranes were incubated overnight at 4 °C with anti-CYP19A1 (sc-130733, 50 kDa) (1:500), anti-FSHR (sc-13935, 75 kDa) (1:500), anti-LHCGR (sc-25828, 85 kDa) (1:250), anti-BCL2 (sc-492, 26 kDa) (1:500) or anti-ACTB (sc-47778, 34 kDa) (1:500) antibody. After wash, secondary antibodies, conjugated to HRP (sc-2004) (1:10 000) were incubated for 2 h with membranes. Protein bands were visualized by incubating the membranes with Immoblot (Millipore Corp., Billerica, MA, USA). The specific bands were recognized regarding the expected sizes on the blot. Band densities were determined and normalized against the beta actin (ACTB) signal using Image J software (NIH, Bethesda, MD, USA). GC culture Isolated GCs were seeded into six-well plates (BD Bioscience, San Jose, CA, USA) at density of 10×105/ml in DMEM/F12 containing 10% FBS, followed by incubation for 8 h at 37 ° with 5% CO2 to allow GC attachment. To remove cell debris and serum factors, cultures were washed twice and incubated in serum-free DMEM/F12, containing 2 mg/ml of HSA (LifeGlobal), 2 mM glutamine (Life Technologies) and 1× Insulin-Transferin-Selenium X (Life Technologies). After overnight culture, medium was replaced once more. GCs were cultured for 1, 3 or 5 days, and medium was changed every 48 h. Cell proliferation and apoptosis assays Cell proliferation analyses were performed by using Vybrant MTT Cell Proliferation Assay Kit (Life Technologies). Briefly, GCs were seeded at densities of 5000 cells/well in 96-well plates. Following a medium change, 10 μl of 12 mM MTT stock solution was added to each well and the plate was incubated at 37 °C for 4 h. Then 100 μl of SDS–HCl was added, and following another 4 h of incubation absorbance of each well was read at 570 nm, using a micro-plate reader (Tecan, Mannedorf, Switzerland). Apoptosis was determined in GCs plated at the density previously described in four-well chamber slides (Thermo Fisher Scientific). Cultured GCs were fixed by adding in 1 ml D-PBS containing 4% paraformaldehyde for 15 min. Slides were then labeled with 10 μg/ml 4,6-diamidino-2-phenylindole-2-HCL (DAPI) (Sigma) for 10 min, and nuclear morphology was assessed using fluorescence microscopy. Apoptotic GCs, exhibiting distinct fragmented nuclei, were counted, and the apoptotic ratio was calculated for each treatment group based on number of apoptotic cells out of a total of 200 cells/slide. Pilot study of early oocyte retrieval Given the results of the previously described experiments, we hypothesized that the oldest patients (Group 3) might benefit clinically if their risk of premature luteinization could be curtailed or completely avoided. Therefore, we reasoned that development of premature luteinization could be pre-empted by scheduling oocyte retrieval earlier. We report here a preliminary summary of such an early retrieval group (ERG), that included 71 consecutive IVF cycles in women above age 43 (mean age 44.8±0.3 years), for which cycle outcomes could be compared to a reference group of 91 women above age 43, who in the preceding year had been treated with normal retrieval timing (NRG, mean age 44.3±0.15 years). The ERG received identical stimulation as previously described for the infertile patients (Groups 2 and 3) and, therefore, identical stimulation to the NRG control group. What distinguished these groups, however, was the timing of hCG administration. While NRG patients had been triggered with hCG at leading follicle size of 19–21 mm, the ERG group was triggered at 16 mm. Otherwise, IVF cycles were identical. Statistical analysis All statistical analyses were performed using Prism software (GraphPad Software, Inc., La Jolla, CA, USA). One-way ANOVA, followed by the Tukey test was used for the statistical analysis of real-time PCR, western blot, MTT assay and cell number calculations. Unpaired t-tests with Welch's correction was used for statistical comparison of clinical data between ERG and NRG patients. The data in all tables and figures are shown as value±s.e.m. Values were considered statistically significant at P<0.05. Results Patient populations and cycle characteristics Patient and IVF cycle characteristics are summarized in Table 1. Mean ages were 24.4±0.52 years for Group 1, 34.1±0.38 years Group 2 and 44.3±0.23 years for Group 3. The table also demonstrates expected increases in FSH and decreases in AMH values with advancing age as well as declining oocyte/embryo numbers and pregnancy rates. Thus, Group 3 clearly reflected the lowest reproductive potential. The table also reports serum P4 to E2 ratios (P4/E2) in all three groups. The P4/E2 was significantly higher in Group 3, while there was no significant difference between Groups 1 and 2. An elevated P4/E2 is a well known marker of premature luteinization (Ozcakir et al. 2004) and, therefore, suggested that older patients might be at higher risk for premature luteinization than the other two groups. Impact of maternal aging on gene expression in human GCs To determine the impact of maternal aging, we quantified expressions of gonadotropin and sex hormone receptors in GCs. FSHR expression was significantly lower in Group 3 patients than Group 1 and Group 2 (Fig. 1A), while, in contrast, LH receptor (LHCGR) expression was significantly higher in Group 3 than Groups 1 and 2 (Fig. 1B). Expressions of estrogen receptor β (ESR2) (Fig. 1C) and androgen receptor (AR) (Fig. 1D) did not differ between the three groups. Down-regulation of FSHR and up-regulation of LHCGR mRNA levels in older patients were then confirmed by western blot (Fig. 2 A, C and D). Figure 1 Download Figure Figure 1 mRNA expression of GC genes was determined by real-time PCR. Values with same letters or without letters above the columns within each unit figure were not different significantly (P>0.05). White columns: Group 1 (oocyte donors), n=7; Grey columns: Group 2 (middle-aged infertile patients), n=10; Black columns: Group 3 (older infertile patients), n=10. Citation: Journal of Endocrinology 226, 3; 10.1530/JOE-15-0246 Figure 2 Download Figure Figure 2 Protein expression of GC genes was determined by western blot. (A) Protein levels of aromatase, FHSR, LHR, BCL-2 and β-actin were evaluated by western blot analysis. G1: Group 1; G2: Group 2; G3: Group 3; (B, C, D, and E) relative quantitative protein levels of aromatase (B), FHSR (C), LHCGR (D), BCL2 (E) by western blot. The experiment was performed four times by using different samples of each age group. White columns: Group 1 (oocyte donors); grey columns: Group 2 (younger infertile patients); black columns: Group 3 (older infertile patients). *P<0.05; **P<0.01. Citation: Journal of Endocrinology 226, 3; 10.1530/JOE-15-0246 To define the steroidogenic activity of GCs with advancing age, mRNA expression of steroidogenic enzymes was analyzed. Aromatase (CYP19A1) (Fig. 1E) and 17β-HSD (HSD17B) (Fig. 1H) were expressed significantly lower in Group 3 women, while P450scc (CYP11A1) (Fig. 1F) was expressed higher. In contrast, expression of the steroidogenic acute regulatory protein (StAR) was similar in all three groups (Fig. 1G). Interestingly, despite small patient numbers, CYP19A1 expression in donors (Group 1) was higher than in both infertility groups (Group 2 and Group 3, P<0.05), a finding confirmed then by western blot (Fig. 2A and B). Because apoptosis is generally increased in older women (Seifer et al. 1996, Sadraie et al. 2000), we also investigated expressions of apoptosis-related genes in GCs (Fig. 1I, J and K). We found no differences in expression of B-cell lymphoma 2 (BCL2), bcl-2-associated X protein (BAX) and survivin (BIRC5) in all groups. These PCR results were then confirmed by western blot (Fig. 2A and E). Combined with increased LHCGR expression, reduced FSHR and CYP19A1 expression in older infertile women further suggests that their GCs undergo earlier luteinization. To obtain further evidence, we also measured expression of progesterone receptor (PGR), another GC differentiation marker. Q-PCR results demonstrated that GCs from older women (Group 3) expressed higher PGR than the other groups (Fig. 1L). Luteinization of GCs, therefore, appears to happen earlier and faster in older women. Impact of maternal aging on proliferation and apoptosis of GCs during in vitro culture To investigate the effect of maternal aging on GC proliferation, we cultured GCs of all three groups in vitro with or without FSH. As Fig. 3A demonstrates, in absence of FSH, cell proliferation between days 1–5 of GCs in Groups 1 and 2 did not change, while in Group 3 patients proliferation declined fast, and to an extremely low level. Distinctively different growing patterns are apparent in Fig. 3C. Figure 3 Download Figure Figure 3 GC proliferation was determined by MTT assay. PI staining after cell culture determined GC apoptosis. (A) Cell proliferation assay was performed after 1–5 days culture. GCs were collected from three different patients in each group and cultured separately. (B) PI staining after 1–5 days culture evaluated GC apoptosis. (C) Distinctively different growing patterns of cultured GCs on days 1–5, with and without FSH supplementation, are apparent in all three age groups. GCs were collected from three different patients in each age group. In the inserted photograph in B, arrows indicate apoptotic cells after PI staining. Values with common letters above the columns within each unit figure were not different significantly (P>0.05). G1: Group 1 (oocyte donors); G2: Group 2 (younger infertile patients); G3: Group 3 (older infertile patients). D1–5: day 1–5 of cell culture; Bar=20 μm. Citation: Journal of Endocrinology 226, 3; 10.1530/JOE-15-0246 Even though we did not observe changed apoptosis-related gene expression in freshly obtained GCs (Fig. 1), we still considered the possibility that the poor cell proliferation we observed in cultured GCs in Group 3 may be caused by increasing apoptosis. As Fig. 3B demonstrates, apoptotic cells increased during culture in all three groups but the increase in apoptotic cells occurred much faster in Group 3. As suggested by others (Tapanainen et al. 1987, Langhout et al. 1991, Rouillier et al. 1998), our results also showed that FSH in all three groups demonstrated positive effects on proliferation and apoptosis of cultured GCs (Fig. 3A and B). In the presence of FSH, GCs from older patients, however, still demonstrate lower proliferation and higher apoptosis after culture. Impact of maternal aging on gene expression of GCs during in vitro culture To determine the effect of age on gene expression of cultured GCs, expression of FSHR, LHCGR, CYP19A1 and BCL2 were investigated by Q-PCR and western blot. As shown in Fig. 4A and C, during culture FSHR and CYP19A1 mRNA expression increased in Groups 1 and 2, but not in Group 3. Presence of FSH in medium enhanced this upregulated expression even in Group 3. Figure 4 Download Figure Figure 4 mRNA expression of GC genes was determined by real-time PCR (A, B, C, and D). Protein expression was determined by western blot. GCs were collected from three different patients in each group and separately cultured. Values with common letters above the columns within each unit figure did not differ significantly (P>0.05). G1, Group 1 (oocyte donors); G2, Group 2 (younger infertile patients); G3, Group 3 (older infertile patients). C, control; FSH, follicle-stimulating hormone. Citation: Journal of Endocrinology 226, 3; 10.1530/JOE-15-0246 PCR results were then confirmed by protein level examination (Fig. 4E). As shown in Fig. 4B, LHCGR expression also increased during culture, though differently from FSHR in that the increase was much faster in Group 3 (Fig. 4B and E). BCL2 expression decreased in all three groups (Fig. 4D), though the fastest in Group 3, with BCL2 protein expression by western blot concurring with PCR results (Fig. 4E). FSH inhibited this decline, suggesting inhibition of apoptosis by FSH in concurrence with previously noted results in Fig. 3B. As demonstrated in Fig. 3B, compared to other groups, we noted higher cell apoptosis in Group 3 after culture in presence of FSH, which did not concur with the observed BCL2 expression in Fig. 4D and E. We, therefore, investigated in addition expressions of two other molecular markers for apoptosis, BAX and BIRC5 by real-time PCR (Supplementary Figure 1, see section on supplementary data given at the end of this article). Since we did not find differences in expression of both of these genes, jointly with our BCL2 findings, this suggests that, though FSH apparently can regulate expression of apoptosis-related genes, it cannot completely reverse apoptosis. Effects of early oocyte retrieval in women of very advanced age (>43 years) A preliminary assessment of early hCG administration in women of advanced age is presented in Table 2. As the table demonstrates, in comparison to 91 historical control cycles in women of very advanced age, who were retrieved with standard timing (the normal retrieval group, NRG), 71 women in this ERG were actually older (44.8±0.3 vs 44.3±0.15 years; P=0.001); Their atretic oocytes were significantly reduced (0.31±0.07 vs 0.78±0.14, P=0.02). Immature oocytes were significantly increased (1.98±0.98 vs 1.1±0.17; P=0.01) but good quality embryos per cycle still significantly increased (3.6±0.36 vs 2.8±0.24; P=0.04). Moreover, clear trends in favor of the ERG were also seen in clinical pregnancy rate per cycle start, clinical pregnancy rate per embryo transfer and in embryo implantation rate, though so far limited patient numbers did not offer the statistical power to reach statistical significance. Table 2 Comparison of IVF cycle outcomes between ERG and NRG ERG (n=71) NRG (n=91) P value Average age (years) 44.8±0.3 44.3±0.15 0.001 Number of follicles/cycle 7.2±0.58 7.3±0.56 0.93 Number of oocytes/cycle 6.7±0.63 5.9±0.49 0.31 Number of immature oocytes 1.98±0.29 1.1±0.17 0.01 Number of atretic oocytes from retrieval/cycle 0.31±0.07 0.78±0.14 0.02 Number of good embryos/cycle 3.6±0.36 2.7±0.24 0.04 Percentage of cycles resulting in pregnancies 15.5 (11/71) 7.7 (7/91) 0.14 Percentage of transferred cycles resulting in pregnancies 19.3 (11/57) 8.9 (7/78) 0.12 Implantation rate (%) 5.3 3.3 0.34 Progesterone/estradiol ratio 1.46±0.16 1.94±0.12 0.002 ERG, early retrieval group; NRG, normal retrieval group. n, patient number. The table, however, also demonstrates that early retrievals significantly decreased P4/E2 ratios in the ERG in comparison to the NRG (1.46±0.16 vs 1.94±0.12, P=0.002). Additionally, Fig. 5 presents gene expression studies, including 12 ERG, 12 NRG and six donor women, demonstrating significant improvements in LHCGR, PGR and CYP19A1 expression of ERG patients, while FSHR expression was not affected significantly (P>0.05) by early retrievals. Figure 5 Download Figure Figure 5 mRNA expression of GC genes was determined by real-time PCR. Values with common letters above the columns within each unit figure were not different significantly (P>0.05). Black columns: older patients with early retrieval, n=3; light grey columns: older patients with normal retrieval n=3; dark grey columns: oocyte donor with normal retrieval (older infertile patients), n=12. Citation: Journal of Endocrinology 226, 3; 10.1530/JOE-15-0246 These preliminary data suggested that early ovulation induction in women of very advanced age demonstrates no adverse outcome effects on IVF, improves a number of well-established outcome parameters of IVF and, likely, ultimately may also improve clinical pregnancy rates in women above age 43, though confirmation of the latter point awaits larger patient numbers. Discussion The decline of female fertility with advancing age is well documented (Tatone 2008, Tatone et al. 2008, Weeg et al. 2012, Younis 2012). It is usually attributed to declining oocyte numbers (Nasseri & Grifo 1998, Out et al. 2000) and quality (Nasseri & Grifo 1998, Slovis & Check 2013). If the assumption of poorer oocyte quality is correct, then even resting follicles and their enclosed oocytes should exhibit the detrimental consequences of ‘aging’. We have questioned this ‘oocentric’ viewpoint on theoretical as well as practical grounds. Since primordial follicles are progenitor structures, widely held to have limited energy needs and metabolic activity, one could alternatively propose that their predisposition toward ‘aging’ is, likely, only minimal. Had they been subject to cumulative damage during natural aging, they only unlikely would have retained the ability to yield pregnancies and normal offspring. Even women of very advanced age and/or with very low functional ovarian reserve, if treated appropriately, however, still conceive and give birth to normal offspring. We, therefore, suggested that the concept of the ‘aging oocyte’ might have to be replaced by a concept of ‘aging ovarian environments’ in which follicles after recruitment undergo growth and maturation (Gleicher et al. 2011a). The difference between these two concepts is fundamental since likelihood of reversing intrinsic aging damage in an ‘aged oocyte’ is practically zero. If the culprit behind ovarian aging is, however, of somatic origin, therapeutic strategies directed towards reconstituting and/or rejuvenating ‘aged ovarian environments’, from which developmentally competent oocytes could be obtained, would offer promise for treatment of age-related infertility in older women. This prospect is supported by androgen-related observations: older women exhibit relative low androgen levels (Gleicher et al. 2013). Moreover, androgens are known to be essential for normal follicle development during early growing follicle stages (Gleicher et al. 2011b). Indeed, androgen supplementation of the ovarian environment by raising testosterone levels improves egg/embryo numbers and quality as well as pregnancy rates (Gleicher & Barad 2011), offering evidence for a novel approach toward therapeutically reversing to a degree selected effects of ovarian ‘aging’. Since GCs surrounding the oocyte define the immediate ovarian microenvironment, their critical role in supporting oocyte development might be the underlying target for endocrine perturbations associated with aging (Buccione et al. 1990). Although numerous studies have documented the relationship between poor oocyte quality and GC abnormalities in human and animals (Buccione et al. 1990, Senbon et al. 2003, Assou et al. 2012, Huang & Wells 2012, Matsuda et al. 2012), effects of age on physiological function and molecular signature of GCs have so far been only sparsely investigated. Indeed, to our knowledge, they have never before been performed in such distinct age groupings (Hurwitz et al. 2010, McReynolds et al. 2012). Our study comprehensively investigated the effects of ovarian aging in humans, and suggests that gene expression, proliferation, apoptosis and ability to respond to FSH stimulation in human GCs are all significantly affected by female age. Notably, Group 3 GCs demonstrated significantly increased LHCGR, PGR and CYP11A1 but reduced FSHR and CYP19A1 expression in comparison to other groups. Similar results have been observed in primates and other species. Luborsky et al. (2002) reported up-regulated LHCGR expression in human luteinized GCs. Increased PGR (Natraj & Richards 1993) and CYP11A1 (Rao et al. 1978) expression has been reported in rat luteinized GCs. Studies in humans and in the bovine suggest that LH surge-induced declines of FSHR represent the initiation of GC luteinization (Nimz et al. 2009, Jeppesen et al. 2012). Likewise, disappearance of CYP19A1 is another marker of luteinization in GCs (Campbell et al. 1998). Our results in older infertility patients (Group 3), therefore, closely parallel previously findings in, and suggest that premature luteinization of GCs is more likely to occur in older than in younger women. High FSH initiates in natural cycles follicular development, leading to rising serum E2 concentrations by CYP19A1. This, in turn, causes a negative feedback on FSH release, and arrests the development of small growing follicles. High concentrations of E2 result in the preovulatory LH surge, which is responsible for final oocyte maturation and ovulation (Laven & Fauser 2006). In here, reported IVF patients, FSH is, however, because of controlled ovarian hyperstimulation, maintained at higher levels. The consequence in some patients can be the triggering of a premature LH surge before follicles/oocytes have fully matured, a phenomenon given the acronym ‘premature luteinization’ (60). Our results are, therefore, consistent with the fact that women above age 43 are at significantly increased risk to develop premature luteinization. That premature luteinization negatively impacts oocyte quality, fertilization and implantation is supported by Skiadas et al. (2012) who demonstrated an association between low functional ovarian reserve in older women and premature luteinization, clinically characterized by higher peripheral LH and lower AMH levels. Also, oocyte numbers and top quality embryo numbers have been reported to be significantly higher in normal patients than in women with premature luteinization (Bosch et al. 2003, Elnashar 2010). In conjunction with here reported molecular data of older women, all of this points to premature luteinization as a principal cause in the age-related decline of female fertility. Higher exogenous FSH exposure may be a contributing factor to the increased risk toward premature luteinization (Elnashar 2010). Women with premature luteinization, indeed, may have higher day 3 FSH levels (Younis et al. 1998, 2001), though there are no data to support higher intracycle levels in the literature. In this study, FSH levels during ovarian hyperstimulation were not higher in Group 3 patients. Their elevated P4/E2 (>1) still suggests an increased risk for premature luteinization (Ozcakir et al. 2004). In light of the widely held notion that physiological luteinization involves GC cell cycle exit and terminal differentiation, our results also suggest that premature luteinization may be linked to GC proliferation arrest and apoptosis. In support, Christenson & Stouffer (1996) reported in primates and rats that an ovulatory luteinizing stimulus causes proliferation arrest and luteinization in cell differentiation (Rao et al. 1978, Oonk et al. 1989, Christenson & Stouffer 1996), findings further supported by the down-regulation of cell cycle proteins, such as p27Kips and cyclin D2 (Fero et al. 1996, Cheng et al. 1999). In cancer cells, cell cycle regulators such as these are well-known mediators, which initiate apoptosis in response to cell cycle arrest (Murphy 2000, Gutierrez et al. 1997). Whether a similar cell cycle checkpoint is operative in GCs of aged women remains to be determined but would be consistent with our observation that GCs of Group 3, indeed, demonstrated a higher level of apoptosis during culture (Fig. 3B). Some caution is, nevertheless, warranted, especially in the presence of serum. GCs in culture spontaneously undergo structural and functional luteinization based upon changes in cell morphology, steroidogenesis and metabolism (Murphy 2000). Although GCs were cultured with serum-free medium here, it is impossible to completely prevent luteinization. FSH supplementation in medium can, inhibit GC luteinization in vitro, as demonstrated in the cow where GC morphology and estrogen production indicate maintenance of a pre-luteinized state (Gutierrez et al. 1997). Similar positive effects of FSH on GC growth and prevention of luteinization were also observed in human and rat (Lambert et al. 2000, Kwintkiewicz et al. 2010, Zhou et al. 2013). The effects of FSH in this study are particularly noteworthy: it significantly enhanced cell proliferation (Fig. 3A and C), reduced apoptosis (Fig. 3B), and up-regulated FSHR (Fig. 4A) and CYP19A1 (Fig. 4C) expression, suggesting at least partial inhibition of luteinization by FSH. Interestingly, although it caused significant improvements, GC function after FSH treatment was still far weaker in GCs of older women (Group 3), when compared to the other two groups, suggesting insufficient FSHR expression. Poor follicular response to FSH in older women is well recognized (Gleicher & Barad 2006). Our observations heightens the significance, which provide compelling evidence that GCs of older women respond less effectively to FSH stimulation during in vitro culture, suggesting an underlying pathophysiology for declining female fertility. Finally, our observation that FSH did not induce LHCGR expression in cultured GCs (Fig. 4B) requires further study because it is generally held that FSH induces LHCGR expression in vivo (Hirakawa et al. 1999, Orisaka et al. 2006, Cannon et al. 2009). A likely explanation for this result is that here investigated GCs were exposed to hCG in vivo. As shown by us and others (Murphy 2000, Hurwitz et al. 2010, Jeppesen et al. 2012, McReynolds et al. 2012), hCG administration causes luteinization of GCs, and a changing physiological and molecular signatures. Therefore, it is possible that hCG administration changes cell sensitivity of the FSH response. This hypothesis is supported by evidence from non-luteinizing GCs, where FSH activates the protein kinase A (PKA) pathway and then induces LHCGR transcription (Oury et al. 1992). But luteinization increases the stability of PKA subunit, which inhibited PKA activation by FSH (Gonzalez-Robayna et al. 1999). That the FSH/PKA-driven transcriptional protein, CREB, undergoes inhibitory phosphorylation in luteinized GCs (78), also supports this hypothesis. Recognizing this pathophysiology in aged GCs then raised the question how such premature luteinization could be prevented. We hypothesized that the likelihood was early oocyte retrieval, which should release oocytes earlier from the hyper-luteinized follicular environments. Preliminary outcome analysis of 71 early retrieval IVF cycles in women above age 43, in comparison to 91 normal routine retrieval cycles, is highly encouraging (Table 2), though much larger patient numbers will be required to unequivocally demonstrate in this patient population that this new management scheme, ultimately, improves IVF pregnancy and delivery rates. This study, however, with considerable certainty established non-inferiority for this new treatment and, with a reasonable level of likelihood suggest that early oocyte retrieval may improve IVF outcomes. The observation that earlier retrieval increased the number of high quality embryos available for transfer by reducing atretic oocyte numbers is reassuring because pregnancy and delivery success in IVF usually follows high quality embryo numbers. Optimism is also warranted since every outcome parameter, which did not significantly improve, without exception, strongly trended in favor of the ERG. It will take at least 150 IVF cycles in this patient population to reach adequate power for final statistical evidence that clinical pregnancy and live birth rates are, indeed, improved by a minimum of 20 percent. In summary, we present here convincing in vivo and in vitro evidence that premature luteinization in infertile women of advanced age was associated with rapidly declining IVF pregnancy rates. We also present preliminary evidence, suggesting that, if such premature luteinization is avoided by earlier oocyte retrieval, IVF outcomes will be improved. Final confirmation of improved pregnancy and delivery outcomes will, however, require a larger patient pool than is available at time of this publication. Supplementary data This is linked to the online version of the paper at http://dx.doi.org/10.1530/JOE-15-0246. Declaration of interest D H B and N G are listed as inventors on a number of US patents, among them, peripherally relevant to this study, patents claiming therapeutic benefits from androgen supplementation of women with LFOR. Both receive royalties from Fertility Nutraceuticals, LLC for licensing of some of these patents. N G holds shares in this company. None of the other authors report any potential conflicts in respect to the study. Funding This work was supported by the Foundation for Reproductive Medicine and intramural grants from the Center for Human Reproduction (CHR) – New York. References AssouSHaouziDDe VosJHamamahS2012Human cumulus cells as biomarkers for embryo and pregnancy outcomes. Molecular Human Reproduction16531–538. (doi:10.1093/molehr/gaq032) Search Google Scholar Export Citation BakerJHardyMPZhouJBondyCLupuFBellveAREfstratiadisA1996Effects of an Igf1 gene null mutation on mouse reproduction. Molecular Endocrinology10903–918. (doi:10.1210/mend.10.7.8813730) Search Google Scholar Export Citation BellocSCohen-BacriePBenkhalifaMCohen-BacrieMDe MouzonJHazoutAMenezoY2008Effect of maternal and paternal age on pregnancy and miscarriage rates after intrauterine insemination. Reproductive Biomedicine Online17392–397. (doi:10.1016/S1472-6483(10)60223-4) Search Google Scholar Export Citation BoschEValenciaIEscuderoECrespoJSimonCRemohiJPellicerA2003Premature luteinization during gonadotropin-releasing hormone antagonist cycles and its relationship with in vitro fertilization outcome. Fertility and Sterility801444–1449. (doi:10.1016/j.fertnstert.2003.07.002) Search Google Scholar Export Citation BrowerPTSchultzRM1982Intercellular communication between granulosa cells and mouse oocytes: existence and possible nutritional role during oocyte growth. Developmental Biology90144–153. (doi:10.1016/0012-1606(82)90219-6) Search Google Scholar Export Citation BuccioneRSchroederACEppigJJ1990Interactions between somatic cells and germ cells throughout mammalian oogenesis. Biology of Reproduction43543–547. (doi:10.1095/biolreprod43.4.543) Search Google Scholar Export Citation CampbellBKBairdDTWebbR1998Effects of dose of LH on androgen production and luteinization of ovine theca cells cultured in a serum-free system. Journal of Reproduction and Fertility11269–77. (doi:10.1530/jrf.0.1120069) Search Google Scholar Export Citation CannonJDSeekalluSVVandevoortCAChaffinCL2009Association of luteinizing hormone receptor gene expression with cell cycle progression in granulosa cells. American Journal of Physiology. Endocrinology and Metabolism296E1392–E1399. (doi:10.1152/ajpendo.90965.2008) Search Google Scholar Export Citation ChengMOlivierPDiehlJAFeroMRousselMFRobertsJMSherrCJ1999The p21(Cip1) and p27(Kip1) CDK ‘inhibitors’ are essential activators of cyclin D-dependent kinases in murine fibroblasts. EMBO Journal181571–1583. (doi:10.1093/emboj/18.6.1571) Search Google Scholar Export Citation ChristensonLKStoufferRL1996Proliferation of microvascular endothelial cells in the primate corpus luteum during the menstrual cycle and simulated early pregnancy. Endocrinology137367–374. (doi:10.1210/endo.137.8536637) Search Google Scholar Export Citation CouseJFYatesMMDerooBJKorachKS2005Estrogen receptor-β is critical to granulosa cell differentiation and the ovulatory response to gonadotropins. Endocrinology1463247–3262. (doi:10.1210/en.2005-0213) Search Google Scholar Export Citation De La FuenteREppigJJ2001Transcriptional activity of the mouse oocyte genome: companion granulosa cells modulate transcription and chromatin remodeling. Developmental Biology229224–236. (doi:10.1006/dbio.2000.9947) Search Google Scholar Export Citation DierichASairamMRMonacoLFimiaGMGansmullerALeMeurMSassone-CorsiP1998Impairing follicle-stimulating hormone (FSH) signaling in vivo: targeted disruption of the FSH receptor leads to aberrant gametogenesis and hormonal imbalance. PNAS9513612–13617. (doi:10.1073/pnas.95.23.13612) Search Google Scholar Export Citation DownsSM1993Purine control of mouse oocyte maturation: evidence that nonmetabolized hypoxanthine maintains meiotic arrest. Molecular Reproduction and Development3582–94. (doi:10.1002/mrd.1080350114) Search Google Scholar Export Citation ElnasharAM2010Progesterone rise on the day of HCG administration (premature luteinization) in IVF: an overdue update. Journal of Assisted Reproduction and Genetics27149–155. (doi:10.1007/s10815-010-9393-8) Search Google Scholar Export Citation FeroMLRivkinMTaschMPorterPCarowCEFirpoEPolyakKTsaiLHBroudyVPerlmutterRM1996A syndrome of multiorgan hyperplasia with features of gigantism, tumorigenesis, and female sterility in p27(Kip1)-deficient mice. Cell85733–744. (doi:10.1016/S0092-8674(00)81239-8) Search Google Scholar Export Citation FisherCRGravesKHParlowAFSimpsonER1998Characterization of mice deficient in aromatase (ArKO) because of targeted disruption of the cyp19 gene. PNAS956965–6970. (doi:10.1073/pnas.95.12.6965) Search Google Scholar Export Citation GleicherNBaradDH2006Effects of transdermal testosterone application on the ovarian response to FSH in poor responders undergoing assisted reproduction technique-a prospective, randomized, double-blind study. Human Reproduction213027; author reply 3027–3028. (doi:10.1093/humrep/del302) Search Google Scholar Export Citation GleicherNBaradDH2011Dehydroepiandrosterone (DHEA) supplementation in diminished ovarian reserve (DOR). Reproductive Biology and Endocrinology967. (doi:10.1186/1477-7827-9-67) Search Google Scholar Export Citation GleicherNWeghoferABaradDH2011aDefining ovarian reserve to better understand ovarian aging. Reproductive Biology and Endocrinology923. (doi:10.1186/1477-7827-9-23) Search Google Scholar Export Citation GleicherNWeghoferABaradDH2011bThe role of androgens in follicle maturation and ovulation induction: friend or foe of infertility treatment?Reproductive Biology and Endocrinology9116. (doi:10.1186/1477-7827-9-116) Search Google Scholar Export Citation GleicherNKimAWeghoferAKushnirVAShohat-TalALazzaroniELeeHJBaradDH2013Hypoandrogenism in association with diminished functional ovarian reserve. Human Reproduction281084–1091. (doi:10.1093/humrep/det033) Search Google Scholar Export Citation Gonzalez-RobaynaIJAllistonTNBusePFirestoneGLRichardsJS1999Functional and subcellular changes in the A-kinase-signaling pathway: relation to aromatase and Sgk expression during the transition of granulosa cells to luteal cells. Molecular Endocrinology131318–1337. (doi:10.1210/mend.13.8.0334) Search Google Scholar Export Citation GrandeMBorrellAGarcia-PosadaRBorobioVMunozMCreusMSolerASanchezABalaschJ2012The effect of maternal age on chromosomal anomaly rate and spectrum in recurrent miscarriage. Human Reproduction273109–3117. (doi:10.1093/humrep/des251) Search Google Scholar Export Citation GutierrezCGCampbellBKWebbR1997Development of a long-term bovine granulosa cell culture system: induction and maintenance of estradiol production, response to follicle-stimulating hormone, and morphological characteristics. Biology of Reproduction56608–616. (doi:10.1095/biolreprod56.3.608) Search Google Scholar Export Citation HerlandsRLSchultzRM1984Regulation of mouse oocyte growth: probable nutritional role for intercellular communication between follicle cells and oocytes in oocyte growth. Journal of Experimental Zoology229317–325. (doi:10.1002/jez.1402290217) Search Google Scholar Export Citation HirakawaTMinegishiTTanoMKamedaTKishiHIbukiYMizutaniTMiyamotoK1999Effect of prolactin on the expression of luteinizing hormone receptors during cell differentiation in cultured rat granulosa cells. Endocrinology1403444–3451. (doi:10.1210/endo.140.8.6888) Search Google Scholar Export Citation HuangZWellsD2012The human oocyte and cumulus cells relationship: new insights from the cumulus cell transcriptome. Molecular Human Reproduction16715–725. (doi:10.1093/molehr/gaq031) Search Google Scholar Export Citation HurwitzJMJindalSGreenseidKBergerDBrooksASantoroNPalL2010Reproductive aging is associated with altered gene expression in human luteinized granulosa cells. Reproductive Sciences1756–67. (doi:10.1177/1933719109348028) Search Google Scholar Export Citation JeppesenJVKristensenSGNielsenMEHumaidanPDal CantoMFadiniRSchmidtKTErnstEYding AndersenC2012LH-receptor gene expression in human granulosa and cumulus cells from antral and preovulatory follicles. Journal of Clinical Endocrinology and Metabolism97E1524–E1531. (doi:10.1210/jc.2012-1427) Search Google Scholar Export Citation KwintkiewiczJNishiYYanaseTGiudiceLC2010Peroxisome proliferator-activated receptor-gamma mediates bisphenol A inhibition of FSH-stimulated IGF-1, aromatase, and estradiol in human granulosa cells. Environmental Health Perspectives118400–406. (doi:10.1289/ehp.0901161) Search Google Scholar Export Citation LambertAHarrisSDKnaggsPRobertsonWR2000Improved FSH sensitisation and aromatase assay in human granulosa-lutein cells. Molecular Human Reproduction6677–680. (doi:10.1093/molehr/6.8.677) Search Google Scholar Export Citation LanghoutDJSpicerLJGeisertRD1991Development of a culture system for bovine granulosa cells: effects of growth hormone, estradiol, and gonadotropins on cell proliferation, steroidogenesis, and protein synthesis. Journal of Animal Science693321–3334. Search Google Scholar Export Citation LavenJSFauserBC2006What role of estrogens in ovarian stimulation. Maturitas54356–362. (doi:10.1016/j.maturitas.2006.04.022) Search Google Scholar Export Citation LuborskyJLThiruppathiPRivnayBRoussevRCoulamCRadwanskaE2002Evidence for different aetiologies of low estradiol response to FSH: age-related accelerated luteinization of follicles or presence of ovarian autoantibodies. Human Reproduction172641–2649. (doi:10.1093/humrep/17.10.2641) Search Google Scholar Export Citation MatsudaFInoueNManabeNOhkuraS2012Follicular growth and atresia in mammalian ovaries: regulation by survival and death of granulosa cells. Journal of Reproduction and Development5844–50. (doi:10.1262/jrd.2011-012) Search Google Scholar Export Citation McReynoldsSDzieciatkowskaMMcCallieBRMitchellSDStevensJHansenKSchoolcraftWBKatz-JaffeMG2012Impact of maternal aging on the molecular signature of human cumulus cells. Fertility and Sterility981574–1580e5. (doi:10.1016/j.fertnstert.2012.08.012) Search Google Scholar Export Citation MehlmannLM2005Stops and starts in mammalian oocytes: recent advances in understanding the regulation of meiotic arrest and oocyte maturation. Reproduction130791–799. (doi:10.1530/rep.1.00793) Search Google Scholar Export Citation MurphyBD2000Models of luteinization. Biology of Reproduction632–11. (doi:10.1095/biolreprod63.1.2) Search Google Scholar Export Citation NasseriAGrifoJA1998Genetics, age, and infertility. Maturitas30189–192. (doi:10.1016/S0378-5122(98)00073-5) Search Google Scholar Export Citation NatrajURichardsJS1993Hormonal regulation, localization, and functional activity of the progesterone receptor in granulosa cells of rat preovulatory follicles. Endocrinology133761–769. (doi:10.1210/endo.133.2.8344215) Search Google Scholar Export Citation NavotDBerghPAWilliamsMAGarrisiGJGuzmanISandlerBGrunfeldL1991Poor oocyte quality rather than implantation failure as a cause of age-related decline in female fertility. Lancet3371375–1377. (doi:10.1016/0140-6736(91)93060-M) Search Google Scholar Export Citation NimzMSpitschakMSchneiderFFurbassRVanselowJ2009Down-regulation of genes encoding steroidogenic enzymes and hormone receptors in late preovulatory follicles of the cow coincides with an accumulation of intrafollicular steroids. Domestic Animal Endocrinology3745–54. (doi:10.1016/j.domaniend.2009.02.002) Search Google Scholar Export Citation van Noord-ZaadstraBMLoomanCWAlsbachHHabbemaJDte VeldeERKarbaatJ1991Delaying childbearing: effect of age on fecundity and outcome of pregnancy. BMJ3021361–1365. (doi:10.1136/bmj.302.6789.1361) Search Google Scholar Export Citation OonkRBKrasnowJSBeattieWGRichardsJS1989Cyclic AMP-dependent and -independent regulation of cholesterol side chain cleavage cytochrome P-450 (P-450scc) in rat ovarian granulosa cells and corpora lutea. cDNA and deduced amino acid sequence of rat P-450scc. Journal of Biological Chemistry26421934–21942. Search Google Scholar Export Citation OrisakaMMizutaniTTajimaKOrisakaSShukunamiKMiyamotoKKotsujiF2006Effects of ovarian theca cells on granulosa cell differentiation during gonadotropin-independent follicular growth in cattle. Molecular Reproduction and Development73737–744. (doi:10.1002/mrd.20246) Search Google Scholar Export Citation OuryFFaucherCRivesIBensaidMBoucheGDarbonJM1992Regulation of cyclic adenosine 3′,5′-monophosphate-dependent protein kinase activity and regulatory subunit RII β content by basic fibroblast growth factor (bFGF) during granulosa cell differentiation: possible implication of protein kinase C in bFGF action. Biology of Reproduction47202–212. (doi:10.1095/biolreprod47.2.202) Search Google Scholar Export Citation OutHJBraatDDLintsenBMGurganTBukulmezOGokmenOKelesGCaballeroPGonzalezJMFabreguesF2000Increasing the daily dose of recombinant follicle stimulating hormone (Puregon) does not compensate for the age-related decline in retrievable oocytes after ovarian stimulation. Human Reproduction1529–35. (doi:10.1093/humrep/15.1.29) Search Google Scholar Export Citation OzcakirHTLeviRTavmergenEGokerEN2004Premature luteinization defined as progesterone estradiol ratio >1 on hCG administration day seems to adversely affect clinical outcome in long gonadotropin-releasing hormone agonist cycles. Journal of Obstetrics and Gynaecology Research30100–104. (doi:10.1111/j.1447-0756.2003.00166.x) Search Google Scholar Export Citation PerreaultSDBarbeeRRSlottVL1988Importance of glutathione in the acquisition and maintenance of sperm nuclear decondensing activity in maturing hamster oocytes. Developmental Biology125181–186. (doi:10.1016/0012-1606(88)90070-X) Search Google Scholar Export Citation RaoMCMidgleyARJrRichardsJS1978Hormonal regulation of ovarian cellular proliferation. Cell1471–78. (doi:10.1016/0092-8674(78)90302-1) Search Google Scholar Export Citation RouillierPMattonPDufourMSirardMAGuilbaultLA1998Steroid production, cell proliferation, and apoptosis in cultured bovine antral and mural granulosa cells: development of an in vitro model to study estradiol production. Molecular Reproduction and Development50170–177. (doi:10.1002/(SICI)1098-2795(199806)50:2%3C;170::AID-MRD7%3E;3.0.CO;2-H) Search Google Scholar Export Citation SadraieSHSaitoHKanekoTSaitoTHiroiM2000Effects of aging on ovarian fecundity in terms of the incidence of apoptotic granulosa cells. Journal of Assisted Reproduction and Genetics17168–173. (doi:10.1023/A:1009422323306) Search Google Scholar Export Citation SeiferDBCharlandCBerlinskyDPenziasASHaningRVJrNaftolinFBarkerBE1993Proliferative index of human luteinized granulosa cells varies as a function of ovarian reserve. American Journal of Obstetrics and Gynecology1691531–1535. (doi:10.1016/0002-9378(93)90430-Q) Search Google Scholar Export Citation SeiferDBGardinerACFerreiraKAPelusoJJ1996Apoptosis as a function of ovarian reserve in women undergoing in vitro fertilization. Fertility and Sterility66593–598. Search Google Scholar Export Citation SeiferDBDeJesusVHubbardK2002Mitochondrial deletions in luteinized granulosa cells as a function of age in women undergoing in vitro fertilization. Fertility and Sterility781046–1048. (doi:10.1016/S0015-0282(02)04214-0) Search Google Scholar Export Citation SenAHammesSR2010Granulosa cell-specific androgen receptors are critical regulators of ovarian development and function. Molecular Endocrinology241393–1403. (doi:10.1210/me.2010-0006) Search Google Scholar Export Citation SenbonSHiraoYMiyanoT2003Interactions between the oocyte and surrounding somatic cells in follicular development: lessons from in vitro culture. Journal of Reproduction and Development49259–269. (doi:10.1262/jrd.49.259) Search Google Scholar Export Citation ShuhaibarLCEgbertJRNorrisRPLampePDNikolaevVOThunemannMWenLFeilRJaffeLA2015Intercellular signaling via cyclic GMP diffusion through gap junctions restarts meiosis in mouse ovarian follicles. PNAS1125527–5532. (doi:10.1073/pnas.1423598112) Search Google Scholar Export Citation SkiadasCCDuanSCorrellMRubioRKaracaNGinsburgESQuackenbushJRacowskyC2012Ovarian reserve status in young women is associated with altered gene expression in membrana granulosa cells. Molecular Human Reproduction18362–371. (doi:10.1093/molehr/gas008) Search Google Scholar Export Citation SlovisBHCheckJH2013Younger women with diminished oocyte reserve are not more prone to meiosis errors leading to spontaneous abortion than their age peers with normal oocyte reserve. Clinical and Experimental Obstetrics & Gynecology4029–32. Search Google Scholar Export Citation TapanainenJLeinonenPJTapanainenPYamamotoMJaffeRB1987Regulation of human granulosa-luteal cell progesterone production and proliferation by gonadotropins and growth factors. Fertility and Sterility48576–580. Search Google Scholar Export Citation TatoneC2008Oocyte senescence: a firm link to age-related female subfertility. Gynecological Endocrinology2459–63. (doi:10.1080/09513590701733504) Search Google Scholar Export Citation TatoneCAmicarelliFCarboneMCMonteleonePCasertaDMarciRArtiniPGPiomboniPFocarelliR2008Cellular and molecular aspects of ovarian follicle ageing. Human Reproduction Update14131–142. (doi:10.1093/humupd/dmm048) Search Google Scholar Export Citation TsujiTKiyosuCAkiyamaKKuniedaT2012CNP/NPR2 signaling maintains oocyte meiotic arrest in early antral follicles and is suppressed by EGFR-mediated signaling in preovulatory follicles. Molecular Reproduction and Development79795–802. (doi:10.1002/mrd.22114) Search Google Scholar Export Citation WebbRJMarshallFSwannKCarrollJ2002Follicle-stimulating hormone induces a gap junction-dependent dynamic change in [cAMP] and protein kinase a in mammalian oocytes. Developmental Biology246441–454. (doi:10.1006/dbio.2002.0630) Search Google Scholar Export Citation WeegNShalom-PazEWiserA2012Age and infertility: the clinical point of view. Minerva Ginecologica64477–483. Search Google Scholar Export Citation WigglesworthKLeeKBO'BrienMJPengJMatzukMMEppigJJ2013Bidirectional communication between oocytes and ovarian follicular somatic cells is required for meiotic arrest of mammalian oocytes. PNAS110E3723–E3729. (doi:10.1073/pnas.1314829110) Search Google Scholar Export Citation YeYKawamuraKSasakiMKawamuraNGroenenPGelpkeMDRauchRHsuehAJTanakaT2009Kit ligand promotes first polar body extrusion of mouse preovulatory oocytes. Reproductive Biology and Endocrinology726. (doi:10.1186/1477-7827-7-26) Search Google Scholar Export Citation YounisJS2012Ovarian aging and implications for fertility female health. Minerva Endocrinologica3741–57. Search Google Scholar Export Citation YounisJSHaddadSMatilskyMBen-AmiM1998Premature luteinization: could it be an early manifestation of low ovarian reserve?Fertility and Sterility69461–465. (doi:10.1016/S0015-0282(97)00561-X) Search Google Scholar Export Citation YounisJSMatilskyMRadinOBen-AmiM2001Increased progesterone/estradiol ratio in the late follicular phase could be related to low ovarian reserve in in vitro fertilization-embryo transfer cycles with a long gonadotropin-releasing hormone agonist. Fertility and Sterility76294–299. (doi:10.1016/S0015-0282(01)01918-5) Search Google Scholar Export Citation ZhangMSuYQSugiuraKXiaGEppigJJ2010Granulosa cell ligand NPPC and its receptor NPR2 maintain meiotic arrest in mouse oocytes. Science330366–369. (doi:10.1126/science.1193573) Search Google Scholar Export Citation ZhouPWuYGLiQLanGCWangGGaoDTanJH2008The interactions between cysteamine, cystine and cumulus cells increase the intracellular glutathione level and developmental capacity of goat cumulus-denuded oocytes. Reproduction135605–611. (doi:10.1530/REP-08-0003) Search Google Scholar Export Citation ZhouPWuYGWeiDLLiQWangGZhangJLuoMJTanJH2010Mouse cumulus-denuded oocytes restore developmental capacity completely when matured with optimal supplementation of cysteamine, cystine, and cumulus cells. Biology of Reproduction82759–768. (doi:10.1095/biolreprod.109.082206) Search Google Scholar Export Citation ZhouPBaumgartenSCWuYBennettJWinstonNHirshfeld-CytronJStoccoC2013IGF-I signaling is essential for FSH stimulation of AKT and steroidogenic genes in granulosa cells. Molecular Endocrinology27511–523. (doi:10.1210/me.2012-1307) Search Google Scholar Export Citation

Elementos de Etnomedicina Veterinaria en la Historia De Venezuela

https://docplayer.es/6533593-Elementos-de-etnomedicina-veterinaria-en-la-historia-de-venezuela.html

Antibiotics - The Perfect Storm

http://antibiotics-theperfectstorm.blogspot.com/2018/12/antibiotics-in-2019.html?m=1 We need new antibiotics to fight infections caused by resistant bacteria. But the marketplace, the structure of the pharmaceutical industry, regulatory agencies, and difficult science are conspiring to deny us the products we need. This blog will present perspectives and developments in the fight for new antibiotics. Wednesday, December 26, 2018 Antibiotics in 2019? Antibiotic R&D has had a particularly bad year starting with The Medicines Company who abandoned their antibiotic R&D efforts and sold their antibiotic assets to Melinta late last year right after getting approval for vabomere. This year both Sanofi and Novartis abandoned their antibiotic R&D efforts and divested their clinical and preclinical assets. Allergan, holder of the North American rights to ceftaroline, dalbavancin and ceftazidime-avibactam, also announced that they would divest their antibiotic assets. I have not heard that they were successful. Achaogen has now undergone two efforts at “restructuring” involving virtually eliminating all R&D and has essentially put up the “for sale” sign just after achieving approval for plazomicin. Finally, Melinta abandoned their antibiotic R&D efforts in the face of miserable sales of their recently launched antibiotics including delafloxacin and vabomere. For the last decade, we have seen the emergence of biotech as the primary antibiotic discovery and development engine replacing the large pharmaceutical companies that have continued to flee the area. Now, it seems, even biotech has hit the brick wall of the broken antibiotic market. Recently, Tetraphase gained approval for eravacycline, a new synthetic tetracycline with activity against resistant pathogens. They have priced the drug such that it will be easier for hospitals to put the drug on their formularies – but their limited label (only for intraabdominal infections) will still limit their sales prospects. Their current market cap is a miserable $58 million. Will Tetraphase achieve a successful launch in the face of all the other antibiotic launch failures? 2019 will provide the answer. Nabriva recently submitted two NDAs for two antibiotics. IV fosfomycin for urinary tract infection is used throughout the world but was never approved in the US. Nabriva purchased Zavante Therapeutics to obtain this asset and filed an NDA for complicated urinary tract infection this year. Lefamulin was discovered and developed by Nabriva (I was involved in getting lefamulin through early development) and targets community acquired pneumonia. Nabriva’s second NDA this year is for lefamulin. Nabriva’s current market cap is $80 million. Once again, 2019 will be the critical year for this emerging biotech company attempting to launch important new antibiotics. What does 2019 have in store for the three large pharmaceutical companies that still maintain antibiotic R&D? This year a new biotech, Prokaryotics, licensed a number of preclinical antibiotic assets from Merck. What antibiotic discovery activity still remains at Merck? I am guessing that this is a minimal effort. At the same time, Merck continues to sell the antibiotics it acquired from Cubist, tedizolid and ceftolozane-tazobactam – but sales have been slow at best. Merck continues its development of imipenem-cilastatin-relebactam at a slow pace. Will Merck continue beyond 2019? The antibiotic R&D effort at GSK has teetered on the edge of the precipice for years. A recent announcement that, with a new CEO, GSK will invest more in oncology and immuno-inflammation while continuing to focus on its top therapy areas: respiratory conditions, HIV and infectious diseases. Announcements like these in the past have not been harbingers of good news for antibiotics researchers. Roche, the third large pharmaceutical company still committed to antibiotics R&D, like Novartis, has its main research site in Basel, Switzerland. But most of the antibiotic interest in the company comes from its Genentech subsidiary in California. Will Roche follow the Novartis example in 2019 or will Genetech convince management to persevere? Without large pharma and their deep pockets, investors are more hesitant to invest in R&D projects. Public-private partnerships like CARB-X and others have, to a certain extent, taken on some of the burden from venture capitalists for antibiotic discovery activity. But even well-funded efforts like CARB-X are likely to come crashing into the sharp rocks of the broken antibiotics market and the lack of big pharma interest. Given the events of the last year and the outlook for new antibiotic launches, I believe 2019 will be the most critical year since most of pharma abandoned antibiotics at the turn of the last century. I fear a catastrophic collapse of antibiotic R&D and commercialization in the absence of government efforts to bolster the broken marketplace for these essential medicines.