The recently created professional science master's degree may be the answer to the increasing need for science-savvy employees in the business world

“I didn't want to be just another MBA,” says Pascal Herzer, one of the first recipients of a new graduate credential known as the professional science master's, or PSM. “Not many people have the ability to understand science and business, and [the PSM] program was designed for that very purpose.”PSMs are two-year American master's degrees financed in large part by the Alfred P. Sloan Foundation to cultivate science managers. Sloan's ultimate goal is to make science careers more attractive to talented young people like Herzer, a 2003 PSM graduate in Applied Biosciences from the University of Arizona, who believes his PSM makes him more marketable to science-based businesses. “I am at the true junction of science and business,” he says.The Missing Degree Fortunately for Herzer, the business of science is booming. Jobs for scientists and engineers grew four times faster than the United States national average since 1980, and should outpace the market until at least 2010. Surprisingly to many academics, most of these jobs are in industry. In 1999, the last year with complete data, two out of three employed science and engineering (S&E) graduates worked in industry, including the great majority of bachelor's and master's degree holders, and 40% of doctorates. In other words, industry, not academe, now drives American S&E employment, and will for the near future. Like academia, industry needs scientifically literate personnel; unlike academia, industry wants employees with business savvy as well. However, in the past, graduate students received either science or business instruction, not both. “Industry simply hired regular master's-degreed people, or MBAs, or more likely PhDs, and just expected them to learn their weaknesses on the job,” says Eleanor L. Babco, Executive Director of the Commission on Professionals in Science and Technology, a nonprofit corporation with funding from the Sloan Foundation to assess PSM graduates. For science-based businesses, then, the American S&E doctorate—viewed by many as the worldwide gold standard for science education—is too specialized for their needs (see  Box 1 ). But a master's degree may be just right. Box 1. Is There a Doctorate in the House? The length of time to obtain a biological science doctorate has increased…
 …The number of postdocs and part-time faculty in the biological sciences is increasing…
 …And the proportion of doctorates in academia is decreasing…
 (Statistics taken from the National Science Board's “Science and Engineering Indicators 2004” [ http://www.nsf.gov/sbe/srs/seind04/ ] and the National Science Foundation Science Resources Statistics Division's “1995 and 2001 Survey of Doctorate Recipients” [ http://www.nsf.gov/sbe/srs/infbrief/nsf04328/table1.xls ].)
The Missing Degree
Fortunately for Herzer, the business of science is booming. Jobs for scientists and engineers grew four times faster than the United States national average since 1980, and should outpace the market until at least 2010. Surprisingly to many academics, most of these jobs are in industry. In 1999, the last year with complete data, two out of three employed science and engineering (S&E) graduates worked in industry, including the great majority of bachelor's and master's degree holders, and 40% of doctorates. In other words, industry, not academe, now drives American S&E employment, and will for the near future.Like academia, industry needs scientifically literate personnel; unlike academia, industry wants employees with business savvy as well. However, in the past, graduate students received either science or business instruction, not both. “Industry simply hired regular master's-degreed people, or MBAs, or more likely PhDs, and just expected them to learn their weaknesses on the job,” says Eleanor L. Babco, Executive Director of the Commission on Professionals in Science and Technology, a nonprofit corporation with funding from the Sloan Foundation to assess PSM graduates.For science-based businesses, then, the American S&E doctorate—viewed by many as the worldwide gold standard for science education—is too specialized for their needs (see  Box 1 ). But a master's degree may be just right.Box 1. Is There a Doctorate in the House? The length of time to obtain a biological science doctorate has increased…
 …The number of postdocs and part-time faculty in the biological sciences is increasing…
 …And the proportion of doctorates in academia is decreasing…
 (Statistics taken from the National Science Board's “Science and Engineering Indicators 2004” [ http://www.nsf.gov/sbe/srs/seind04/ ] and the National Science Foundation Science Resources Statistics Division's “1995 and 2001 Survey of Doctorate Recipients” [ http://www.nsf.gov/sbe/srs/infbrief/nsf04328/table1.xls ].)
Box 1. Is There a Doctorate in the House?
The length of time to obtain a biological science doctorate has increased……The number of postdocs and part-time faculty in the biological sciences is increasing……And the proportion of doctorates in academia is decreasing…(Statistics taken from the National Science Board's “Science and Engineering Indicators 2004” [ http://www.nsf.gov/sbe/srs/seind04/ ] and the National Science Foundation Science Resources Statistics Division's “1995 and 2001 Survey of Doctorate Recipients” [ http://www.nsf.gov/sbe/srs/infbrief/nsf04328/table1.xls ].)Bridging the Gap During the 20th century, the master's degree evolved as a professional credential in many fields, including business, education, and social work, and more recently, pharmacy, physical therapy, and accounting. In the 1990s, non-incidental master's in the sciences—in other words, intentionally terminal degrees, not consolation prizes for failing out of graduate school—crept into engineering and applied mathematics, too, as companies grew more reliant on computational analysis and hired accordingly. From 1981 to 2000, for example, the number of earned master's degrees in mathematics and computer science more than doubled. With hopes of spurring a “significant movement,” in 1997 the Sloan Foundation bet big on professional master's degrees, eventually spending $11 million on almost 100 programs across the US. Sloan Foundation–backed PSM programs now operate at 45 universities in 20 states, in such fields as microbial biotechnology and applied genomics; similar programs have also developed independently of the Sloan Foundation, such as the Master of Science in Bioinformatics at Johns Hopkins. And while most PSM-style programs are currently in the US, this may soon change: the 1999 “Bologna Agreement” requires all European Union universities to adopt uniform undergraduate and graduate degrees “relevant to the European labour market”; so master's-level industry-centric degrees are sure to follow. At Leiden University in the Netherlands, for example, students can now add a “science-based business” focus to any research master of science (MSc) program. Like all graduate programs, PSMs offer advanced coursework in a (science or math) specialization, usually in an emerging or hybrid field such as bioinformatics. Most PSMs also provide business courses—including finance, project management, regulatory affairs, and intellectual property law—and information technology classes as well. PSMs are “industry relevant” by design, with external advisory committees populated by local business leaders, weekly colloquia led by corporate representatives, special arrangements for employed students, and industry internships or final projects exploring realistic business scenarios (see  Box 2 ). Box 2. Requirements for PSM Programs Only programs meeting most of the following requirements may earn the official moniker “professional science master's.”
 Two years of science or math graduate-level coursework, taught by regular faculty, characterized by interdisciplinary studies and a focus on informatics Training in business fundamentals—such as finance, marketing, project management, communication, and team building—and exposure to industry professionals Final project reflecting a realistic workplace issue and/or industry internship Advisory board of industry professionals Targeted recruitment and admissions separate from other degree programs Commitment to tracking graduates through first five years Long-term sustainability
 (Source:  http://www.sciencemasters.com/affiliation.html .) A key principle underlying the PSM model is interdisciplinarity. PSM students are encouraged to reach out to other departments and broaden their expertise in multiple areas, to better understand the collaborative culture of industry-style scientific enterprise. To promote such connections, PSM programs explicitly teach teamwork and effective scientific communication, with authentic case studies analyzed alongside MBA students, classroom presentations and public seminars, and open defenses of final projects. Consequently, PSM graduates, unlike many doctoral graduates, are trained to possess a wide array of interactive skills, including sizing up an audience for their ability to comprehend the presented material and adapting appropriately. In a science-based business, ideas must flow freely between scientists and non-scientists in and out of the company—between researchers and marketers, say, or inventors and patent lawyers—to capitalize on discoveries and comply with regulations. When non-scientists misunderstand the science underpinning a business model, profits suffer. But the presence of a central employee who streams data between differently educated members of the network may boost the bottom line. PSM students are specifically trained to act as such “science translators.” “[My PSM] allows me to serve as an efficient mediator between corporate entities, university personnel, and scientists,” says Herzer. For this reason, small companies and start-ups, which cannot afford specialists for every position, may particularly benefit from PSM-credentialed employees, able to connect different people and function in multiple roles; indeed, many PSM graduates have job descriptions expressly created for them. “We need generalists rather than specialists,” says James L. Ratcliff, Chairman and CEO of Rowpar Pharmaceuticals, a dental products company in Scottsdale, Arizona. For small companies like his, Ratcliff says, “PSM graduates have an appropriate combination of project management expertise, an understanding of business environments and priorities, and advanced knowledge in the physical and life sciences.” Although it is too early for comprehensive assessment, employment outcomes for PSM graduates have been examined, and this result is clear: they are getting industry jobs. According to The Conference Board, an independent business management organization funded by the Sloan Foundation to survey PSM alumni, by 2002, 91% of the first PSM graduates had obtained full-time positions within their field despite a white-collar recession, two-thirds with salaries of $50,000 or more. A separate analysis by the Commission on Professionals in Science and Technology found that 61.5% of employed respondents were hired by businesses. Employment opportunities range from marketing to bioinformatics (see  Box 3 ). “Companies need people that can work in companies,” says Lindy A. Brigham, coordinator of the Applied Biosciences PSM program at the University of Arizona. Box 3. First Jobs Obtained by PSM Graduates A sampling of actual first jobs obtained by students in PSM or PSM-style programs:
 Coordinator of Regulatory Affairs Associate Criminalist Licensing Assistant Staff Researcher Senior Computer Database Specialist Project Manager E-Product Marketing Specialist Clinical Consultant Technical Support Specialist Bioinformatics Programmer Manager of Medical Affairs
 (Source: personal communications from L. A. Brigham, D. Ascher, T. Tiongson Pohar, and S. Inamdar.)
Bridging the Gap
During the 20th century, the master's degree evolved as a professional credential in many fields, including business, education, and social work, and more recently, pharmacy, physical therapy, and accounting. In the 1990s, non-incidental master's in the sciences—in other words, intentionally terminal degrees, not consolation prizes for failing out of graduate school—crept into engineering and applied mathematics, too, as companies grew more reliant on computational analysis and hired accordingly. From 1981 to 2000, for example, the number of earned master's degrees in mathematics and computer science more than doubled.With hopes of spurring a “significant movement,” in 1997 the Sloan Foundation bet big on professional master's degrees, eventually spending $11 million on almost 100 programs across the US. Sloan Foundation–backed PSM programs now operate at 45 universities in 20 states, in such fields as microbial biotechnology and applied genomics; similar programs have also developed independently of the Sloan Foundation, such as the Master of Science in Bioinformatics at Johns Hopkins. And while most PSM-style programs are currently in the US, this may soon change: the 1999 “Bologna Agreement” requires all European Union universities to adopt uniform undergraduate and graduate degrees “relevant to the European labour market”; so master's-level industry-centric degrees are sure to follow. At Leiden University in the Netherlands, for example, students can now add a “science-based business” focus to any research master of science (MSc) program.Like all graduate programs, PSMs offer advanced coursework in a (science or math) specialization, usually in an emerging or hybrid field such as bioinformatics. Most PSMs also provide business courses—including finance, project management, regulatory affairs, and intellectual property law—and information technology classes as well. PSMs are “industry relevant” by design, with external advisory committees populated by local business leaders, weekly colloquia led by corporate representatives, special arrangements for employed students, and industry internships or final projects exploring realistic business scenarios (see  Box 2 ).Box 2. Requirements for PSM Programs Only programs meeting most of the following requirements may earn the official moniker “professional science master's.”
 Two years of science or math graduate-level coursework, taught by regular faculty, characterized by interdisciplinary studies and a focus on informatics Training in business fundamentals—such as finance, marketing, project management, communication, and team building—and exposure to industry professionals Final project reflecting a realistic workplace issue and/or industry internship Advisory board of industry professionals Targeted recruitment and admissions separate from other degree programs Commitment to tracking graduates through first five years Long-term sustainability
 (Source:  http://www.sciencemasters.com/affiliation.html .)
Box 2. Requirements for PSM Programs
Only programs meeting most of the following requirements may earn the official moniker “professional science master's.”Two years of science or math graduate-level coursework, taught by regular faculty, characterized by interdisciplinary studies and a focus on informatics Training in business fundamentals—such as finance, marketing, project management, communication, and team building—and exposure to industry professionals Final project reflecting a realistic workplace issue and/or industry internship Advisory board of industry professionals Targeted recruitment and admissions separate from other degree programs Commitment to tracking graduates through first five years Long-term sustainabilityTwo years of science or math graduate-level coursework, taught by regular faculty, characterized by interdisciplinary studies and a focus on informaticsTraining in business fundamentals—such as finance, marketing, project management, communication, and team building—and exposure to industry professionalsFinal project reflecting a realistic workplace issue and/or industry internshipAdvisory board of industry professionalsTargeted recruitment and admissions separate from other degree programsCommitment to tracking graduates through first five yearsLong-term sustainability(Source:  http://www.sciencemasters.com/affiliation.html .)A key principle underlying the PSM model is interdisciplinarity. PSM students are encouraged to reach out to other departments and broaden their expertise in multiple areas, to better understand the collaborative culture of industry-style scientific enterprise. To promote such connections, PSM programs explicitly teach teamwork and effective scientific communication, with authentic case studies analyzed alongside MBA students, classroom presentations and public seminars, and open defenses of final projects. Consequently, PSM graduates, unlike many doctoral graduates, are trained to possess a wide array of interactive skills, including sizing up an audience for their ability to comprehend the presented material and adapting appropriately.In a science-based business, ideas must flow freely between scientists and non-scientists in and out of the company—between researchers and marketers, say, or inventors and patent lawyers—to capitalize on discoveries and comply with regulations. When non-scientists misunderstand the science underpinning a business model, profits suffer. But the presence of a central employee who streams data between differently educated members of the network may boost the bottom line. PSM students are specifically trained to act as such “science translators.” “[My PSM] allows me to serve as an efficient mediator between corporate entities, university personnel, and scientists,” says Herzer.For this reason, small companies and start-ups, which cannot afford specialists for every position, may particularly benefit from PSM-credentialed employees, able to connect different people and function in multiple roles; indeed, many PSM graduates have job descriptions expressly created for them. “We need generalists rather than specialists,” says James L. Ratcliff, Chairman and CEO of Rowpar Pharmaceuticals, a dental products company in Scottsdale, Arizona. For small companies like his, Ratcliff says, “PSM graduates have an appropriate combination of project management expertise, an understanding of business environments and priorities, and advanced knowledge in the physical and life sciences.”Although it is too early for comprehensive assessment, employment outcomes for PSM graduates have been examined, and this result is clear: they are getting industry jobs. According to The Conference Board, an independent business management organization funded by the Sloan Foundation to survey PSM alumni, by 2002, 91% of the first PSM graduates had obtained full-time positions within their field despite a white-collar recession, two-thirds with salaries of $50,000 or more. A separate analysis by the Commission on Professionals in Science and Technology found that 61.5% of employed respondents were hired by businesses. Employment opportunities range from marketing to bioinformatics (see  Box 3 ). “Companies need people that can work in companies,” says Lindy A. Brigham, coordinator of the Applied Biosciences PSM program at the University of Arizona.Box 3. First Jobs Obtained by PSM Graduates A sampling of actual first jobs obtained by students in PSM or PSM-style programs:
 Coordinator of Regulatory Affairs Associate Criminalist Licensing Assistant Staff Researcher Senior Computer Database Specialist Project Manager E-Product Marketing Specialist Clinical Consultant Technical Support Specialist Bioinformatics Programmer Manager of Medical Affairs
 (Source: personal communications from L. A. Brigham, D. Ascher, T. Tiongson Pohar, and S. Inamdar.)
Box 3. First Jobs Obtained by PSM Graduates
A sampling of actual first jobs obtained by students in PSM or PSM-style programs:
 Coordinator of Regulatory Affairs Associate Criminalist Licensing Assistant Staff Researcher Senior Computer Database Specialist Project Manager E-Product Marketing Specialist Clinical Consultant Technical Support Specialist Bioinformatics Programmer Manager of Medical AffairsCoordinator of Regulatory AffairsAssociate CriminalistLicensing AssistantStaff ResearcherSenior Computer Database SpecialistProject ManagerE-Product Marketing SpecialistClinical ConsultantTechnical Support SpecialistBioinformatics ProgrammerManager of Medical Affairs(Source: personal communications from L. A. Brigham, D. Ascher, T. Tiongson Pohar, and S. Inamdar.)Not a Perfect Cure Although most scientific careers demand a graduate degree, a professional master's in many hard sciences still encounters entrenched academic opposition. According to Lee-Jen Wei, then acting chair of the Department of Biostatistics in the Harvard School of Public Health, quoted in the  Wall Street Journal , “Harvard tries to create leadership in industry, academics and government, and our philosophy is we don't think that with a master's degree people can fill that role very easily.” The government appears to agree with this view. While most doctoral candidates receive federal funds for tuition and other expenses, there is little money for master's students, who disproportionately end up in industry regardless of specialization. PSM students are especially affected by this problem because “interdisciplinary” equals “expensive.” Similarly, “interdisciplinary” can also mean “hard to find”—companies with targeted recruitment often miss PSM students, who are not “in” any particular department—and “confusing”—differences in these new, still somewhat vaguely defined programs can make hiring comparisons difficult. But perhaps the most conspicuous drawback to PSMs is their newness, and resulting obscurity: almost half of graduates say they are “not sure” employers will value their PSM, or the unique skill set it affords.
Not a Perfect Cure
Although most scientific careers demand a graduate degree, a professional master's in many hard sciences still encounters entrenched academic opposition. According to Lee-Jen Wei, then acting chair of the Department of Biostatistics in the Harvard School of Public Health, quoted in the  Wall Street Journal , “Harvard tries to create leadership in industry, academics and government, and our philosophy is we don't think that with a master's degree people can fill that role very easily.”Wall Street JournalThe government appears to agree with this view. While most doctoral candidates receive federal funds for tuition and other expenses, there is little money for master's students, who disproportionately end up in industry regardless of specialization. PSM students are especially affected by this problem because “interdisciplinary” equals “expensive.” Similarly, “interdisciplinary” can also mean “hard to find”—companies with targeted recruitment often miss PSM students, who are not “in” any particular department—and “confusing”—differences in these new, still somewhat vaguely defined programs can make hiring comparisons difficult. But perhaps the most conspicuous drawback to PSMs is their newness, and resulting obscurity: almost half of graduates say they are “not sure” employers will value their PSM, or the unique skill set it affords.But Will They Succeed? Still, many observers of higher education support the PSM concept. Judith Glazer-Raymo, author of the forthcoming book  Professionalizing Graduate Education: The Master's Degree in the Marketplace , argues that converging market forces will lead to the success of the professional master's degree in science. These forces include: rapid technological change; the rise of alternative learning channels such as online and distance education, corporate universities, and hi-tech certification programs; the proliferation of degrees in general, and in multidisciplinary fields specifically; and a fundamental societal shift away from public service and toward entrepreneurship, profitability, and competition. Kenneth R. Smith, former dean of the Eller College of Business and Public Administration at the University of Arizona, and others make the case that PSMs may protect students' careers from outsourcing to foreign countries. The American S&E labor pool is shrinking, and industry has already responded by transferring much of its research and development overseas; however, companies are mostly moving lab scientists, not strategic analysts. Cross-training in both science and business could thus provide an edge for domestic workers in the near-term employment environment; in fact, PSM programs have a higher proportion of US citizens and residents than S&E doctoral programs. Further, in its 2003 report, the National Science Board urged the government to better align S&E graduate education with “expected national skill needs,” including “interdisciplinary skills.” The report also recommended federal funding for a wider range of educational options and more attention on the real economic concerns of students—code words for support of professional master's degree initiatives. In the same vein, top universities now advocate “interconnections” between their professional schools and traditional departments, as a way of strengthening the overall academic mission, and many countries are sponsoring initiatives to stimulate university–industry links, to maximize marketing of technological innovations. For advocates, then, the PSM both advances the cause of science education reform and addresses changing employment conditions with one big idea: reinvention of the two-year graduate credential for an entrepreneurial age. Herzer, for one, now a technology development representative at the Scripps Research Institute, has staked his future on the potential of professional master's degrees. “Scientists rarely understand business dealings, and business personnel rarely comprehend scientific discoveries,” he says. “The overlay of the two is crucial for any successful business transaction of scientific origins.” Median length of time to doctorate degree Number of doctorates employed in the biological sciences, by position Science and engineering doctorates by employment sector
But Will They Succeed?
Still, many observers of higher education support the PSM concept. Judith Glazer-Raymo, author of the forthcoming book  Professionalizing Graduate Education: The Master's Degree in the Marketplace , argues that converging market forces will lead to the success of the professional master's degree in science. These forces include: rapid technological change; the rise of alternative learning channels such as online and distance education, corporate universities, and hi-tech certification programs; the proliferation of degrees in general, and in multidisciplinary fields specifically; and a fundamental societal shift away from public service and toward entrepreneurship, profitability, and competition.Professionalizing Graduate Education: The Master's Degree in the MarketplaceKenneth R. Smith, former dean of the Eller College of Business and Public Administration at the University of Arizona, and others make the case that PSMs may protect students' careers from outsourcing to foreign countries. The American S&E labor pool is shrinking, and industry has already responded by transferring much of its research and development overseas; however, companies are mostly moving lab scientists, not strategic analysts. Cross-training in both science and business could thus provide an edge for domestic workers in the near-term employment environment; in fact, PSM programs have a higher proportion of US citizens and residents than S&E doctoral programs.Further, in its 2003 report, the National Science Board urged the government to better align S&E graduate education with “expected national skill needs,” including “interdisciplinary skills.” The report also recommended federal funding for a wider range of educational options and more attention on the real economic concerns of students—code words for support of professional master's degree initiatives. In the same vein, top universities now advocate “interconnections” between their professional schools and traditional departments, as a way of strengthening the overall academic mission, and many countries are sponsoring initiatives to stimulate university–industry links, to maximize marketing of technological innovations.For advocates, then, the PSM both advances the cause of science education reform and addresses changing employment conditions with one big idea: reinvention of the two-year graduate credential for an entrepreneurial age. Herzer, for one, now a technology development representative at the Scripps Research Institute, has staked his future on the potential of professional master's degrees. “Scientists rarely understand business dealings, and business personnel rarely comprehend scientific discoveries,” he says. “The overlay of the two is crucial for any successful business transaction of scientific origins.”


Background Premature Ovarian Failure (POF), defined as the development of hypergonadotropic amenorrhea before the age of 40 years, occurs in about 1% of all women. Other than karyotype abnormalities, very few genes are known to be associated with this ovarian dysfunction. Recently, in seven patients who presented with POF and white matter abnormalities on MRI (ovarioleukodystrophy) eight mutationswere found in  EIF2B2 ,  4  and  5 .
Background
Premature Ovarian Failure (POF), defined as the development of hypergonadotropic amenorrhea before the age of 40 years, occurs in about 1% of all women. Other than karyotype abnormalities, very few genes are known to be associated with this ovarian dysfunction. Recently, in seven patients who presented with POF and white matter abnormalities on MRI (ovarioleukodystrophy) eight mutationswere found in  EIF2B2 ,  4  and  5 .EIF2B245Methods To further test the involvement of known mutations of  EIF2B  genes in POF, we screened 93 patients with POF who did not have identified leukodystrophy or neurological symptoms. We evaluated these eight mutations and two additional mutations that had been found in patients with milder forms of eIF2B-related disorders. We used restriction enzymes and direct sequencing.
Methods
To further test the involvement of known mutations of  EIF2B  genes in POF, we screened 93 patients with POF who did not have identified leukodystrophy or neurological symptoms. We evaluated these eight mutations and two additional mutations that had been found in patients with milder forms of eIF2B-related disorders. We used restriction enzymes and direct sequencing.EIF2BResults None of the known mutations in  EIF2B  genes, either homozygous or heterozygous, were identified in our 93 patients with pure 46,XX POF. The upper 95 % confidence limit of the proportion 0/93 is 3.2%.
Results
None of the known mutations in  EIF2B  genes, either homozygous or heterozygous, were identified in our 93 patients with pure 46,XX POF. The upper 95 % confidence limit of the proportion 0/93 is 3.2%.EIF2BConclusions We conclude that eIF2B mutations, already described in cases of POF associated with white matter abnormalities, are an uncommon cause of pure spontaneous premature ovarian failure.
Conclusions
We conclude that eIF2B mutations, already described in cases of POF associated with white matter abnormalities, are an uncommon cause of pure spontaneous premature ovarian failure.

Background Premature Ovarian Failure (POF) can present as a primary or secondary amenorrhea and is associated with elevated gonadotropins before 40 years of age. POF affects 1% of all women and occurs in 0.1% before the age of 30 years [ 1 ]. POF has been associated with karyotype abnormalities, including various X chromosome aberrations such as Turner syndrome, which causes depletion of ovarian follicles during development [ 2 ]. While conditions such as autoimmune diseases are also associated with POF, the cause is unknown in about 90 % of cases. However, since many affected women have a family history of the condition, predisposition to POF may be inherited [ 3 ]. To date, mutations associated with POF have been identified in a small number of genes [ 4 ], including those encoding the inhibin alpha [ 5 ], the FSH receptor [ 6 ], the LH/choriogonadotrophin receptor [ 7 ], and the forkhead transcription factor 2 [ 8 ]. No more than 10% of women with ovarian failure have mutations in these different genes [ 8 ]. Recently three of the five  EIF2B  genes ( EIF2B2 ,  4  and  5 ) were reportedly involved in seven patients who presented with POF and white matter abnormalities on MRI (ovarioleukodystrophy) [ 9 ]. These genes encode the five subunits of the eucaryotic initiation factor 2B (eIF2B alpha to epsilon), which is involved in the first step of protein synthesis. eIF2B-related disorders include a large group of phenotypes with a recognizable MRI pattern but different clinical severities. The clinical  spectrum  can range from a rapid course leading to death in severe congenital forms to asymptomatic MRI findings in adult patients [ 10 , 11 ]. Ovarioleukodystrophy might present in a phase without neurological symptoms and an apparently isolated form of POF [ 9 ]. Therefore, we screened a series of 93 patients with apparently pure, karyotypically normal POF for mutations in  EIF2B  genes.
Background
Premature Ovarian Failure (POF) can present as a primary or secondary amenorrhea and is associated with elevated gonadotropins before 40 years of age. POF affects 1% of all women and occurs in 0.1% before the age of 30 years [ 1 ]. POF has been associated with karyotype abnormalities, including various X chromosome aberrations such as Turner syndrome, which causes depletion of ovarian follicles during development [ 2 ]. While conditions such as autoimmune diseases are also associated with POF, the cause is unknown in about 90 % of cases. However, since many affected women have a family history of the condition, predisposition to POF may be inherited [ 3 ]. To date, mutations associated with POF have been identified in a small number of genes [ 4 ], including those encoding the inhibin alpha [ 5 ], the FSH receptor [ 6 ], the LH/choriogonadotrophin receptor [ 7 ], and the forkhead transcription factor 2 [ 8 ]. No more than 10% of women with ovarian failure have mutations in these different genes [ 8 ].Recently three of the five  EIF2B  genes ( EIF2B2 ,  4  and  5 ) were reportedly involved in seven patients who presented with POF and white matter abnormalities on MRI (ovarioleukodystrophy) [ 9 ]. These genes encode the five subunits of the eucaryotic initiation factor 2B (eIF2B alpha to epsilon), which is involved in the first step of protein synthesis. eIF2B-related disorders include a large group of phenotypes with a recognizable MRI pattern but different clinical severities. The clinical  spectrum  can range from a rapid course leading to death in severe congenital forms to asymptomatic MRI findings in adult patients [ 10 , 11 ]. Ovarioleukodystrophy might present in a phase without neurological symptoms and an apparently isolated form of POF [ 9 ]. Therefore, we screened a series of 93 patients with apparently pure, karyotypically normal POF for mutations in  EIF2B  genes.EIF2BEIF2B245spectrumEIF2BMethods Selection of patients with premature ovarian failure In the current study, we evaluated the presence of  EIF2B  mutations in 93 unrelated and well-characterized women with POF. An institutional review board approved the study and all participants gave a written informed consent. Referring physicians made the diagnosis of premature ovarian failure based on the following criteria: development of at least 4 months of amenorrhea before age 40 associated with two serum FSH levels in the menopausal range. Women with premature ovarian failure as a result of surgery, radiation, chemotherapy, or known karyotype abnormalities were not included in the study. There were 6 Asians, 12 Blacks, 4 Hispanics and 71 Caucasians. The median age at the onset of menstrual irregularity was 24.5 years (range 13 to 39). Eighteen women had a family history of POF. All women underwent a history and physical examination and laboratory screening to confirm the diagnosis of POF and all had a normal karyotype. None of the women had evidence of a neurological disorder. EIF2B mutations screening Genomic DNA was extracted from peripheral blood using standard procedures. The exons of the genes  EIF2B2 ,  4  and  5  which contain mutations found in POF patients or in milder forms of eIF2B-related disorders were amplified by the polymerase chain reaction (PCR) as previously described (Table  1 )[ 11 ]. Table 1 Sequences of PCR primers used and their PCR conditions. Nucleotide change tested (gene) Primers sequences (5'-3') PCR conditions C512T ( EIF2B2 ), C547T ( EIF2B2 ) F: GCAAAACCGTTCTTAC R: CCTACCCATCTCTCGTTTAT PCR preparation : 1.5 mM MgCl2, 0.225 mM dNTP, 0.8 μM primers, 100 ng primers, 1 unit AmpliTaq Gold™(Applied Biosystems), 1X Taq buffer. 607–612del/insTG ( EIF2B2 ), A638G ( EIF2B2 ) F: GGAAATTATGTGCTGGATATG R: ACTTTATTCTCTCACCGTGGAT P243L ( EIF2B4 ) F: ATGCTCAAGCTCCCTTTCAA R: CTTCACAACTTACAAAGCCT R374C ( EIF2B4 ) F: ATTCAAGCACCTGGCATGAT R: CGCTGCACTCCATCCTTATC PCR reaction : 95°C 12 min, 35 cycles (94°C 30 s, 55°C 30 s, 72°C 45 s), 72°C 10 min, 4°C. T1393C ( EIF2B4 ), T1465C ( EIF2B4 ) F: TGTCCTGTAAGTAGGGGACCTT R: AAGGGGTTGTGAAGTCTGGA G338A ( EIF2B5 ), C583T ( EIF2B5 ) F: GAGAAGGACTGTGAGTGCTGA R: GCCTTCTAAGGGGACAATAAC F: forward primer, R: reverse primer Nine mutations were tested by restriction enzymes directly on PCR products (Table  2 ): 500 ng of PCR products were incubated with 1 unit of specific restriction enzyme from Biolabs ®  Inc. for 90 minutes, according to the supplier's instructions. Restriction fragments were analyzed by standard acrylamide gel electrophoresis. Table 2 EIF2B2 ,  4  and  5  mutations tested with restriction enzymes. Mutation tested: nucleotides changes (amino acid changes) Mutated gene Restriction enzyme used PCR product size (base pairs) Restriction profile (number of restriction fragments: their size in base pairs) No mut* Het mut* Hom mut* C512T (S171F) EIF2B2 Hpy188III 251 4 fragments: 51, 29, 16 and 155 bp. 5 fragments: 51, 29, 16, 155 and 171 bp. 3 fragments: 51, 29 and 171 bp. 607–612del/insTG (M203fs) EIF2B2 HphI 313 2 fragments: 310 and 3 bp. 4 fragments: 310, 142, 164 and 3 bp. 3 fragments: 142, 164 and 3 bp. C547T (R183stop) EIF2B2 EcoNI 253 2 fragments: 120 and 133 bp. 4 fragments: 120, 133, 14 and 119 bp. 3 fragments: 120, 14 and 119 bp. A638G (E213G) EIF2B2 BsmAI 313 2 fragments: 153 and 160 bp. 3 fragments: 313, 160 and 153 bp. 1 fragment: 313 bp P243L (C728T) EIF2B4 AciI 612 3 fragments: 353, 223 and 36 bp. 4 fragments: 576, 353, 223 and 36 bp. 2 fragments: 576 and 36 bp. R374C (C1120T) EIF2B4 HpyCH4IV 640 2 fragments: 447 and 193 bp. 3 fragments: 640, 447 and 193 bp. 1 fragment: 640 bp. T1393C (C465R) EIF2B4 BsrDI 694 3 fragments: 368, 32 and 294 bp. 4 fragments: 368, 32, 294 and 326 bp. 2 fragments: 368 and 326 bp. T1465C (Y489H) EIF2B4 NlaIII 707 3 fragments: 109, 310 and 288 bp. 5 fragments: 109, 310, 288, 57 and 231 bp. 4 fragments: 109, 310, 57 and 231 bp. G338A (R113H) EIF2B5 Fnu4HI 800 3 fragments: 115, 633 and 52 bp. 4 fragments: 115, 633, 52 and 748 bp. 2 fragments: 748 and 52 bp. * No mut: no mutation; Het mut: heterozygous mutation; Hom mut: homozygous mutation. The C583T (R195C) mutation in the  EIF2B5  gene was tested by direct sequencing of exon 4 as previously described [ 11 ].
Methods
Selection of patients with premature ovarian failure In the current study, we evaluated the presence of  EIF2B  mutations in 93 unrelated and well-characterized women with POF. An institutional review board approved the study and all participants gave a written informed consent. Referring physicians made the diagnosis of premature ovarian failure based on the following criteria: development of at least 4 months of amenorrhea before age 40 associated with two serum FSH levels in the menopausal range. Women with premature ovarian failure as a result of surgery, radiation, chemotherapy, or known karyotype abnormalities were not included in the study. There were 6 Asians, 12 Blacks, 4 Hispanics and 71 Caucasians. The median age at the onset of menstrual irregularity was 24.5 years (range 13 to 39). Eighteen women had a family history of POF. All women underwent a history and physical examination and laboratory screening to confirm the diagnosis of POF and all had a normal karyotype. None of the women had evidence of a neurological disorder.
Selection of patients with premature ovarian failure
In the current study, we evaluated the presence of  EIF2B  mutations in 93 unrelated and well-characterized women with POF. An institutional review board approved the study and all participants gave a written informed consent. Referring physicians made the diagnosis of premature ovarian failure based on the following criteria: development of at least 4 months of amenorrhea before age 40 associated with two serum FSH levels in the menopausal range. Women with premature ovarian failure as a result of surgery, radiation, chemotherapy, or known karyotype abnormalities were not included in the study. There were 6 Asians, 12 Blacks, 4 Hispanics and 71 Caucasians. The median age at the onset of menstrual irregularity was 24.5 years (range 13 to 39). Eighteen women had a family history of POF. All women underwent a history and physical examination and laboratory screening to confirm the diagnosis of POF and all had a normal karyotype. None of the women had evidence of a neurological disorder.EIF2BEIF2B mutations screening Genomic DNA was extracted from peripheral blood using standard procedures. The exons of the genes  EIF2B2 ,  4  and  5  which contain mutations found in POF patients or in milder forms of eIF2B-related disorders were amplified by the polymerase chain reaction (PCR) as previously described (Table  1 )[ 11 ]. Table 1 Sequences of PCR primers used and their PCR conditions. Nucleotide change tested (gene) Primers sequences (5'-3') PCR conditions C512T ( EIF2B2 ), C547T ( EIF2B2 ) F: GCAAAACCGTTCTTAC R: CCTACCCATCTCTCGTTTAT PCR preparation : 1.5 mM MgCl2, 0.225 mM dNTP, 0.8 μM primers, 100 ng primers, 1 unit AmpliTaq Gold™(Applied Biosystems), 1X Taq buffer. 607–612del/insTG ( EIF2B2 ), A638G ( EIF2B2 ) F: GGAAATTATGTGCTGGATATG R: ACTTTATTCTCTCACCGTGGAT P243L ( EIF2B4 ) F: ATGCTCAAGCTCCCTTTCAA R: CTTCACAACTTACAAAGCCT R374C ( EIF2B4 ) F: ATTCAAGCACCTGGCATGAT R: CGCTGCACTCCATCCTTATC PCR reaction : 95°C 12 min, 35 cycles (94°C 30 s, 55°C 30 s, 72°C 45 s), 72°C 10 min, 4°C. T1393C ( EIF2B4 ), T1465C ( EIF2B4 ) F: TGTCCTGTAAGTAGGGGACCTT R: AAGGGGTTGTGAAGTCTGGA G338A ( EIF2B5 ), C583T ( EIF2B5 ) F: GAGAAGGACTGTGAGTGCTGA R: GCCTTCTAAGGGGACAATAAC F: forward primer, R: reverse primer Nine mutations were tested by restriction enzymes directly on PCR products (Table  2 ): 500 ng of PCR products were incubated with 1 unit of specific restriction enzyme from Biolabs ®  Inc. for 90 minutes, according to the supplier's instructions. Restriction fragments were analyzed by standard acrylamide gel electrophoresis. Table 2 EIF2B2 ,  4  and  5  mutations tested with restriction enzymes. Mutation tested: nucleotides changes (amino acid changes) Mutated gene Restriction enzyme used PCR product size (base pairs) Restriction profile (number of restriction fragments: their size in base pairs) No mut* Het mut* Hom mut* C512T (S171F) EIF2B2 Hpy188III 251 4 fragments: 51, 29, 16 and 155 bp. 5 fragments: 51, 29, 16, 155 and 171 bp. 3 fragments: 51, 29 and 171 bp. 607–612del/insTG (M203fs) EIF2B2 HphI 313 2 fragments: 310 and 3 bp. 4 fragments: 310, 142, 164 and 3 bp. 3 fragments: 142, 164 and 3 bp. C547T (R183stop) EIF2B2 EcoNI 253 2 fragments: 120 and 133 bp. 4 fragments: 120, 133, 14 and 119 bp. 3 fragments: 120, 14 and 119 bp. A638G (E213G) EIF2B2 BsmAI 313 2 fragments: 153 and 160 bp. 3 fragments: 313, 160 and 153 bp. 1 fragment: 313 bp P243L (C728T) EIF2B4 AciI 612 3 fragments: 353, 223 and 36 bp. 4 fragments: 576, 353, 223 and 36 bp. 2 fragments: 576 and 36 bp. R374C (C1120T) EIF2B4 HpyCH4IV 640 2 fragments: 447 and 193 bp. 3 fragments: 640, 447 and 193 bp. 1 fragment: 640 bp. T1393C (C465R) EIF2B4 BsrDI 694 3 fragments: 368, 32 and 294 bp. 4 fragments: 368, 32, 294 and 326 bp. 2 fragments: 368 and 326 bp. T1465C (Y489H) EIF2B4 NlaIII 707 3 fragments: 109, 310 and 288 bp. 5 fragments: 109, 310, 288, 57 and 231 bp. 4 fragments: 109, 310, 57 and 231 bp. G338A (R113H) EIF2B5 Fnu4HI 800 3 fragments: 115, 633 and 52 bp. 4 fragments: 115, 633, 52 and 748 bp. 2 fragments: 748 and 52 bp. * No mut: no mutation; Het mut: heterozygous mutation; Hom mut: homozygous mutation. The C583T (R195C) mutation in the  EIF2B5  gene was tested by direct sequencing of exon 4 as previously described [ 11 ].
EIF2B mutations screening
Genomic DNA was extracted from peripheral blood using standard procedures.The exons of the genes  EIF2B2 ,  4  and  5  which contain mutations found in POF patients or in milder forms of eIF2B-related disorders were amplified by the polymerase chain reaction (PCR) as previously described (Table  1 )[ 11 ].EIF2B245Sequences of PCR primers used and their PCR conditions.EIF2B2EIF2B2EIF2B2EIF2B2EIF2B4EIF2B4EIF2B4EIF2B4EIF2B5EIF2B5F: forward primer, R: reverse primerNine mutations were tested by restriction enzymes directly on PCR products (Table  2 ): 500 ng of PCR products were incubated with 1 unit of specific restriction enzyme from Biolabs ®  Inc. for 90 minutes, according to the supplier's instructions. Restriction fragments were analyzed by standard acrylamide gel electrophoresis.EIF2B2 ,  4  and  5  mutations tested with restriction enzymes.EIF2B245EIF2B2EIF2B2EIF2B2EIF2B2EIF2B4EIF2B4EIF2B4EIF2B4EIF2B5* No mut: no mutation; Het mut: heterozygous mutation; Hom mut: homozygous mutation.The C583T (R195C) mutation in the  EIF2B5  gene was tested by direct sequencing of exon 4 as previously described [ 11 ].EIF2B5Results None of the eight mutations already described in ovarioleukodystrophy were detected in our 93 patients with pure 46,XX POF, neither in a homozygous nor in a heterozygous state. In addition, the mutations C728T and C1120T ( EIF2B4 ) described in milder forms of eIF2B-related disorders were not found in this series of 93 patients with POF. The upper 95 % confidence limit of the proportion 0/93 is 3.2%.
Results
None of the eight mutations already described in ovarioleukodystrophy were detected in our 93 patients with pure 46,XX POF, neither in a homozygous nor in a heterozygous state. In addition, the mutations C728T and C1120T ( EIF2B4 ) described in milder forms of eIF2B-related disorders were not found in this series of 93 patients with POF. The upper 95 % confidence limit of the proportion 0/93 is 3.2%.EIF2B4Discussion eIF2B-related disorders include a large group of phenotypes with different clinical severities. Individuals can be classified into three clinical groups according to their age at disease onset: <2 years (group 1), 2 to 5 years (group 2) and > 5 years (group 3) [ 11 ]. Group 3 corresponds to individuals with the milder form of the disease, including the six families (seven patients) already described presenting with ovarioleukodystrophy [ 9 ]. In these six eIF2B-mutated families, neurological symptoms with abnormalities of the cerebral white matter on MRI were associated with primary or secondary amenorrhea due to POF [ 9 ]. A correlation was observed between the age of onset of the neurological deterioration and the severity of the ovarian failure, suggesting a common pathophysiological pathway [ 9 ]. The mutated eIF2B may be responsible for both increased apoptosis of ovarian follicles leading to POF, and a defect in glial cell development causing abnormal formation of white matter structures. In ovarioleukodystrophy, a phase of amenorrhea without neurological symptoms can be observed, suggesting that an apparently isolated case of POF might be due to  EIF2B  mutations. In the present study, we tested for  EIF2B  mutations a series of 93 patients with pure, karyotypically normal POF without identified signs of cerebral dysfunction. In eIF2B-related disorders, a correlation exists between genotype and disease onset [ 11 ]. The mutations G338A ( EIF2B5  gene) and A638G ( EIF2B2  gene) are found in 71% of families with late onset forms of eIF2B-related disorders (group 3) [ 11 ]. In ovarioleukodystrophy, 4/6 families have a G338A or A638G mutation in a heterozygote or a homozygote state. Thus, to further evaluate involvement of eIF2B mutations in apparently isolated cases of POF, we restricted our screening to the 10 mutations associated with the late onset form (group 3) of eIF2B-related disorders. In the present series of 93 patients with pure, karyotypically normal POF, no mutations were detected, suggesting a low frequency of  EIF2B  mutations in women with POF who have no apparent neurological signs.
Discussion
eIF2B-related disorders include a large group of phenotypes with different clinical severities. Individuals can be classified into three clinical groups according to their age at disease onset: <2 years (group 1), 2 to 5 years (group 2) and > 5 years (group 3) [ 11 ]. Group 3 corresponds to individuals with the milder form of the disease, including the six families (seven patients) already described presenting with ovarioleukodystrophy [ 9 ].In these six eIF2B-mutated families, neurological symptoms with abnormalities of the cerebral white matter on MRI were associated with primary or secondary amenorrhea due to POF [ 9 ]. A correlation was observed between the age of onset of the neurological deterioration and the severity of the ovarian failure, suggesting a common pathophysiological pathway [ 9 ].The mutated eIF2B may be responsible for both increased apoptosis of ovarian follicles leading to POF, and a defect in glial cell development causing abnormal formation of white matter structures. In ovarioleukodystrophy, a phase of amenorrhea without neurological symptoms can be observed, suggesting that an apparently isolated case of POF might be due to  EIF2B  mutations. In the present study, we tested for  EIF2B  mutations a series of 93 patients with pure, karyotypically normal POF without identified signs of cerebral dysfunction.EIF2BEIF2BIn eIF2B-related disorders, a correlation exists between genotype and disease onset [ 11 ]. The mutations G338A ( EIF2B5  gene) and A638G ( EIF2B2  gene) are found in 71% of families with late onset forms of eIF2B-related disorders (group 3) [ 11 ]. In ovarioleukodystrophy, 4/6 families have a G338A or A638G mutation in a heterozygote or a homozygote state. Thus, to further evaluate involvement of eIF2B mutations in apparently isolated cases of POF, we restricted our screening to the 10 mutations associated with the late onset form (group 3) of eIF2B-related disorders. In the present series of 93 patients with pure, karyotypically normal POF, no mutations were detected, suggesting a low frequency of  EIF2B  mutations in women with POF who have no apparent neurological signs.EIF2B5EIF2B2EIF2BConclusions For patients presenting with POF without neurological signs or MRI abnormalities, the routine screening of the  EIF2B  mutations is not clinically indicated.
Conclusions
For patients presenting with POF without neurological signs or MRI abnormalities, the routine screening of the  EIF2B  mutations is not clinically indicated.EIF2BCompeting interests The author(s) declare that they have no competing interests.
Competing interests
The author(s) declare that they have no competing interests.Authors'contributions AF and FGB carried out the molecular genetics studies, including enzyme restrictions (AF) and sequencing (AF and FGB). AF drafted and conceived of the study. RS participated in the coordination of the study. VHV, VKB, LMN recruited and evaluated the patients, collected DNA samples, participated in the design and coordination of the study, and helped in drafting the manuscript. OBT conceived of the study, and participated in its design and coordination. All authors participated in the writing of the manuscript and have read and approved the final manuscript.
Authors'contributions
AF and FGB carried out the molecular genetics studies, including enzyme restrictions (AF) and sequencing (AF and FGB). AF drafted and conceived of the study. RS participated in the coordination of the study. VHV, VKB, LMN recruited and evaluated the patients, collected DNA samples, participated in the design and coordination of the study, and helped in drafting the manuscript. OBT conceived of the study, and participated in its design and coordination. All authors participated in the writing of the manuscript and have read and approved the final manuscript.Pre-publication history The pre-publication history for this paper can be accessed here:
Pre-publication history
The pre-publication history for this paper can be accessed here: