Educational guidelines on sexual orientation, gender identity and expression, and sex characteristics biases in medical education.

A commitment to diversity, equity, inclusion, and belonging in medical education requires addressing both explicit and implicit biases based on sexual orientation, gender identity and expression, and sex characteristics and the intersectionality with other identities. Heterosexism and heteronormative attitudes contribute to health and healthcare disparities for lesbian, gay, bisexual, transgender and queer or questioning, intersex, asexual individuals. Student, trainee, and faculty competencies in medical education curricula regarding the care of lesbian, gay, bisexual, transgender and queer or questioning, intersex, asexual patients and those who are gender nonconforming or born with differences of sex development allow for better understanding and belonging within the clinical learning environment of lesbian, gay, bisexual, transgender and queer/questioning, intersex, asexual learners and educators. The Association of Professors of Gynecology and Obstetrics issued a call to action to achieve a future free from racism and bias through inclusivity in obstetrics and gynecology education and healthcare, which led to the creation of the Association of Professors of Gynecology and Obstetrics Diversity, Equity, and Inclusion Guidelines Task Force. The task force initially addressed racism, racial- and ethnicity-based bias, and discrimination in medical education and additionally identified other groups that are subject to bias and discrimination, including sexual orientation, gender identity and expression, and sex characteristic identities, persons with disabilities, and individuals with various religious and spiritual practices. In this scholarly perspective, the authors expand on previously developed guidelines to address sexual orientation, gender identity and expression, and sex characteristics bias, heterosexism, and heteronormative attitudes in obstetrics and gynecology educational products, materials, and clinical learning environments to improve access and equitable care to vulnerable individuals of the lesbian, gay, bisexual, transgender and queer or questioning, intersex, asexual community.

Educational guidelines for diversity and inclusion: addressing racism and eliminating biases in medical education.

Racism and bias contribute to healthcare disparities at a patient and population health level and also contribute to the stagnation or even regression of progress toward equitable representation in the workforce and in healthcare leadership. Medical education and healthcare systems have expended tremendous efforts over the past several years to address these inequities. However, systemic racism continues to impact health outcomes and the future physician workforce. The Association of Professors of Gynecology and Obstetrics called for action to achieve a future free from racism in obstetrics and gynecology education and healthcare. As a result of this call to action, the Diversity, Equity, and Inclusion Guidelines Task Force was created. The mission of the Task Force was to support educators in their efforts to identify and create educational materials that augment antiracist educational goals and prepare, recruit, and retain a talented and diverse workforce. In this Special Report, the authors share these guidelines that describe best practices and set new standards to increase diversity, foster inclusivity, address systemic racism, and eliminate bias in obstetrics and gynecology educational products, materials, and environments.

Obstetrics and gynecology resident perception of virtual fellowship interviews.

BACKGROUND: Travel restrictions amidst the COVID-19 pandemic reshaped interviewing for fellowships into a predominantly virtual process. How this impacts Obstetrics and Gynecology (OB/GYN) resident approaches to fellowship application and Match navigation is largely unknown. METHODS: We performed a cross-sectional survey study of fourth year OB/GYN residents in the United States who participated in at least one virtual fellowship interview in 2020. We collected information regarding demographics, application strategy, perceived strengths and weaknesses of virtual interviews, and confidence with rank list creation. Descriptive statistics were used for categorical variables and responses pre- and post-Match were compared using Fisher's exact test. RESULTS: Seventy-five out of an estimated 490 applicants (~ 15% response rate) completed the survey. Of the respondents, 65.3% felt they interviewed at more programs virtually than they would anticipate completing in person, but perceived less confidence in having the necessary information (n = 45, 60%) or understanding the culture of programs (n = 59, 78.7%) to create a rank list. Cost savings were the main benefit of virtual interviews (n = 50, 66.7%), and inability to get a true "feel" for a program was the biggest limitation (n = 43, 57.3%). A majority (46.7%) advocate for a future hybrid interview process. CONCLUSIONS: OB/GYN residents pursuing fellowship reported interviewing at more programs during the virtual season, but had less confidence with rank list creation. Cost savings benefits are weighed against difficulty getting a "feel" for programs virtually. Most would advocate for a future hybrid interview process.

To the Point: advising students applying to Obstetrics and Gynecology residency in 2020 and beyond.

This article, from the "To the Point" series by the Undergraduate Medical Education Committee of the Association of Professors of Gynecology and Obstetrics, is a guide for advising medical students applying to Obstetrics and Gynecology residency programs. The residency application process is changing rapidly in response to an increasingly complex and competitive atmosphere, with a wider recognition of the stress, expense, and difficulty of matching into graduate training programs. The coronavirus disease 2019 pandemic and societal upheaval make this application cycle more challenging than ever before. Medical students need reliable, accurate, and honest advising from the faculty in their field of choice to apply successfully to residency. The authors outline a model for faculty career advisors, distinct from mentors or general academic advisors. The faculty career advisor has detailed knowledge about the field, an in-depth understanding of the application process, and what constitutes a strong application. The faculty career advisor provides accurate information regarding residency programs within the specialty, helping students to strategically apply to programs where the student is likely to match, decreasing anxiety, expense, and overapplication. Faculty career advisor teams advise students throughout the application process with periodic review of student portfolios and are available for support and advice throughout the process. The authors provide a guide for the faculty career advisor in Obstetrics and Gynecology, including faculty development and quality improvement.

CREOGs Over Coffee: Feasibility of an Ob-Gyn Medical Education Podcast by Residents.

BACKGROUND: Podcasts and other digital resources are increasingly popular among medical learners and allow the dissemination of research to larger audiences. Little is known about the feasibility of graduate medical education trainees developing podcasts for their own and others' learning. OBJECTIVE: We described the development and implementation of a medical education podcast series by residents for obstetrics and gynecology (Ob-Gyn) resident learning, and demonstrated feasibility, sustainability, and acceptance of this series. METHODS: We used the Council on Resident Education in Obstetrics and Gynecology (CREOG) educational guidelines to create a weekly study podcast for Ob-Gyn residents over 10 months. Costs and donations (for feasibility), downloads over time (for sustainability), and number of reviews on Apple iTunes and followers on Twitter (for acceptability) were measured. RESULTS: Sixty episodes were released from September 30, 2018, to July 28, 2019 (43 weeks). Initial costs included $3,150 startup and $29 monthly. Online donations through Patreon amounted to $200 a month, which covered 58% of startup costs at 10 months and are projected to cover full costs by 1.5 years. The podcast had 173 995 downloads as recorded through Podbean (39 a month in September, increased to 31 206 a month in July). It gained 644 followers on Twitter and 147 ratings on iTunes, with an average of 4.86 out of 5 stars. CONCLUSIONS: Medical podcasts created by Ob-Gyn residents during their training appear feasible and highly acceptable over a sustained period.

Randomized crossover study investigating resident retention of menopause-related knowledge after completion of learning modules.

OBJECTIVE: The aim of this study was to evaluate residents' retention of menopause-based knowledge immediately after, and 3 months after completion of, self-administered modules that varied by menopause-related topic and delivery format. METHODS: Prospective crossover study of Obstetrics and Gynecology and Family Medicine residents at one institution over the 2017 to 2018 academic year. Residents were randomized to a series of three PowerPoints (Microsoft, Redwood, WA), each <30 slides, administered during regularly scheduled didactics. Each series contained three subjects (Menopause Basics [MB], Hormone Therapy [HT], and Genitourinary Syndrome of Menopause [GSM]) delivered through three different presentation styles (typical presentation [typical], pictures and a narration [pictures], and interactive to reveal information [interactive]). Knowledge and comfort were assessed through baseline, immediate postexposure, and 3-month follow-up surveys containing 24 knowledge questions (multiple choice) and 10 comfort and satisfaction questions (5-point Likert scale and multiple choice). Statistical tests were applied with P<0.05 considered significant. RESULTS: Thirty-three residents completed the 3-month follow-up. Immediately postexposure, knowledge and comfort increased from baseline for all topics (P < 0.05). When formats were grouped together to investigate retention by topic, the HT topic demonstrated a sustained increase in knowledge on 3-month follow-up (P = 0.047). The typical format of the GSM topic had significantly better retention than the picture format (P = 0.027). All formats were associated with a significant increase in comfort (all P < 0.01). CONCLUSIONS: Participation in this specialized menopause curriculum led to short-term increases in objectively assessed menopause-related knowledge. Tailoring self-administered learning modules to learning styles did not, however, effectively enhance overall knowledge retention on 3-month follow-up, though comfort in managing menopause remained increased. : Video Summary:http://links.lww.com/MENO/A474.

Obstetric Fistula Repair in Sub-Saharan Africa: Partnering to Create Sustainable Impact for Patients and Trainees.

Obstetric fistula, an abnormal connection between a woman's genital tract and urinary tract or rectum, can be physically and psychosocially debilitating. We describe a sustainable obstetric fistula surgical trip model that includes providers from Women and Infants Hospital at Brown University. These surgical trips provide pre-operative, surgical, and post-operative care to patients with fistulae at Kibagabaga Hospital in Kigali, Rwanda. To ensure patients are prepared for the recovery process after fistula surgery, the team created a post-operative education curriculum that includes illustrative visual aids and teaching guides translated into Kinyarwanda, focusing on topics including urinary catheter care, wound care, and pain management. Through this program, the team is committed to restoring women's dignity through fistula repair as well as providing a model for delivery of sustainable surgical care in low-resource settings. Involvement of trainees into a global health team like this can benefit both the trainee and the patients served.

Impact of 4 Components of Instructional Design Video on Medical Student Medical Decision Making During the Inpatient Rounding Experience.

INTRODUCTION: The Four Components of Instructional Design (4C-ID) Model has been used to teach Medical Decision Making (MDM), a core competency recognized by the Liaison Committee for Medical Education. 4 Components of Instructional Design (4C-ID) has been applied in general medical education, but not the inpatient clerkship setting. A 4C-ID video for inpatient rounding, like postpartum rounding in Ob/Gyn, could help improve MDM on busy services. METHODS: Students in the third year Ob/Gyn clerkship were randomized by clerkship group to receive a 20-minute postpartum rounding video, based on 4C-ID principles, or usual teaching. MDM and knowledge were assessed pre-/postintervention with the Diagnostic Thinking Inventory and a case-based evaluation. Satisfaction was assessed with Likert style questions. RESULTS: Seventy-eight students were randomized (36 control, 42 intervention). Both groups had equal baseline measures of MDM and knowledge, and similar postclerkship MDM. The intervention group demonstrated higher knowledge postclerkship (17.1, 22.6 p < 0.001). Students in the intervention felt prepared by the video, and would recommend it. Students in the control group reported higher satisfaction with their postpartum rounding experience (3.9, 3.5 p = 0.04). DISCUSSION: Videos are easy to incorporate teaching platforms for medical students, however, the 4C-ID based video in this study did not increase student MDM. In addition, educators should use caution when integrating video into coursework as use of video may lead to decreased student satisfaction as it did in this study.

Global Health Interest Among Female Pelvic Medicine and Reconstructive Surgery Fellows.

OBJECTIVE: To determine interest in global health (GH) work among Female Pelvic Medicine and Reconstructive Surgery (FPMRS)fellows. STUDY DESIGN: An anonymous, online survey was sent to FPMRS fellows in March 2012. All fellows at accredited and nonaccredited U.S. FPMRS programs were eligible. Of at least 123 fellows, 58 (47%) completed the survey and met inclusion criteria. Survey questions included demographics, GH interest and experience, barriers to GH experience, and career goals. RESULTS: Of those 58 fellows, 79% of respondents graduated from Ob-Gyn residencies, 41% were first year fellows, 45% spoke another language fluently, and 62% had previously worked and/or studied in a developing country. Of the respondents 74% desired GH experience during fellowship, 78% desired GH experience after fellowship, and 40% reported seeing themselves integrating GH into their career. Top barriers to GH work in fellowship were lack of elective time (74%), cost (70%), and personal commitments (67%). A total of 39% of respondents said the ability to work in GH somewhat or strongly affected their decision to pursue FPMRS, and 26% said the availability of GH opportunities affected their fellowship rank list. Family (88%), clinical commitments (78%), and cost (67%) were the biggest reported hurdles to future GH work. CONCLUSION: Nearly three-quarters of FPMRS fellows are interested in GH work in fellowship. Almost half would like to include it in future practice. Barriers in fellowship include elective time, cost, and personal commitments.

The ABRF Edman Sequencing Research Group 2008 Study: investigation into homopolymeric amino acid N-terminal sequence tags and their effects on automated Edman degradation.

The Edman Sequence Research Group (ESRG) of the Association of Biomolecular Resource designs and executes interlaboratory studies investigating the use of automated Edman degradation for protein and peptide analysis. In 2008, the ESRG enlisted the help of core sequencing facilities to investigate the effects of a repeating amino acid tag at the N-terminus of a protein. Commonly, to facilitate protein purification, an affinity tag containing a polyhistidine sequence is conjugated to the N-terminus of the protein. After expression, polyhistidine-tagged protein is readily purified via chelation with an immobilized metal affinity resin. The addition of the polyhistidine tag presents unique challenges for the determination of protein identity using Edman degradation chemistry. Participating laboratories were asked to sequence one protein engineered in three configurations: with an N-terminal polyhistidine tag; with an N-terminal polyalanine tag; or with no tag. Study participants were asked to return a data file containing the uncorrected amino acid picomole yields for the first 17 cycles. Initial and repetitive yield (R.Y.) information and the amount of lag were evaluated. Information about instrumentation and sample treatment was also collected as part of the study. For this study, the majority of participating laboratories successfully called the amino acid sequence for 17 cycles for all three test proteins. In general, laboratories found it more difficult to call the sequence containing the polyhistidine tag. Lag was observed earlier and more consistently with the polyhistidine-tagged protein than the polyalanine-tagged protein. Histidine yields were significantly less than the alanine yields in the tag portion of each analysis. The polyhistidine and polyalanine protein-R.Y. calculations were found to be equivalent. These calculations showed that the nontagged portion from each protein was equivalent. The terminal histidines from the tagged portion of the protein were demonstrated to be responsible for the high lag during N-terminal sequence analysis.

ABRF ESRG 2006 study: Edman sequencing as a method for polypeptide quantitation.

The Edman Sequencing Research Group (ESRG) designs studies on the use of Edman degradation for protein and peptide analysis. These studies provide a means for participating laboratories to compare their analyses against a benchmark of those from other laboratories that provide this valuable service. The main purpose of the 2006 study was to determine how accurate Edman sequencing is for quantitative analysis of polypeptides. Secondarily, participants were asked to identify a modified amino acid residue, N-epsilon-acetyl lysine [Lys(Ac)], present within one of the peptides. The ESRG 2006 peptide mixture consisted of three synthetic peptides. The Peptide Standards Research Group (PSRG) provided two peptides, with the following sequences: KAQYARSVLLEKDAEPDILELATGYR (peptide B), and RQAKVLLYSGR (peptide C). The third peptide, peptide C*, synthesized and characterized by ESRG, was identical to peptide C but with acetyl lysine in position 4. The mixture consisted of 20% peptide B and 40% each of peptide C and its acetylated form, peptide C*. Participating laboratories were provided with two tubes, each containing 100 picomoles of the peptide mixture (as determined by quantitative amino acid analysis) and were asked to provide amino acid assignments, peak areas, retention times at each cycle, as well as initial and repetitive yield estimates for each peptide in the mixture. Details about instruments and parameters used in the analysis were also collected. Participants in the study with access to a mass spectrometer (MALDI-TOF or ESI) were asked to provide information about the relative peak areas of the peptides in the mixture as a comparison with the peptide quantitation results from Edman sequencing. Positive amino acid assignments were 88% correct for peptide C and 93% correct for peptide B. The absolute initial sequencing yields were an average of 67% for peptide (C+C*) and 65.6 % for peptide B. The relative molar ratios determined by Edman sequencing were an average of 4.27 (expected ratio of 4) for peptides (C+C*)/B, and 1.49 for peptide C*/C (expected ratio of 1); the seemingly high 49% error in quantification of Lys(Ac) in peptide C* can be attributed to commercial unavailability of its PTH standard. These values compare very favorably with the values obtained by mass spectrometry.

Factors useful in predicting the success of oocyte donation: a 3-year retrospective analysis.

OBJECTIVE: To establish prognostic relevance of parameters assessed in oocyte donation cycles. DESIGN: Retrospective analysis. SETTING: Large university-based donor oocyte program. PATIENT(S): All oocyte recipient cycles achieving embryo transfer from September 1995 to October 1998. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Pregnancy. RESULT(S): Recipient age and reproductive status, day 9 and 12 serum estradiol (E(2)) levels and a progesterone (P) level obtained 2 days after initiation of hormonal therapy did not correlate with pregnancy. Endometrial thickness, but not endometrial pattern, was useful in predicting pregnancy outcome. The clinical pregnancy and live-birth rate in cycles where the endometrial thickness was less than 8 mm was significantly lower when compared to cycles with an endometrial thickness > or =9 mm. Cycles where optimal quality embryos were transferred had the highest implantation (36%), clinical pregnancy (63%) and live birth (54%) rates and these rates were significantly higher than those of cycles where only poor quality embryos were available for transfer (10% implantation, 17% clinical pregnancy, and 8% live birth rates, respectively; P<.05). CONCLUSION(S): The most reliable predictive factors for pregnancy in oocyte donation cycles are the quality of the embryos transferred and the recipient's mid-cycle endometrial thickness. Recipient monitoring should minimally include ultrasound assessment of endometrial thickness.

Construction and biological characterization of an HB-GAM/FGF-1 chimera for vascular tissue engineering.

OBJECTIVE: Cardiovascular tissue engineering approaches to vessel wall restoration have focused on the potent but relatively nonspecific and heparin-dependent mesenchymal cell mitogen fibroblast growth factor 1 (FGF-1). We hypothesized that linking FGF-1 to a sequence likely to bind to cell surface receptors relatively more abundant on endothelial cells (ECs) might induce a relative greater EC bioavailability of the FGF-1. We constructed a heparin-binding growth-associated molecule (HB-GAM)/FGF-1 chimera by linking full-length human HB-GAM to the amino-terminus of human FGF-1beta (21-154) and tested its activities on smooth muscle cells (SMCs) and ECs. METHODS: Primary canine carotid SMCs and jugular vein ECs were plated in 96-well plates in media containing 10% fetal bovine serum and grown to approximately 80% confluence. After being growth arrested in serum-free media for 24 hours, the cells were exposed to concentration ranges of cytokines and heparin, and proliferation was measured with tritiated-thymidine incorporation. Twenty percent fetal bovine serum was used as positive control, and phosphate-buffered saline was used as negative control. RESULTS: In the presence of heparin the HB-GAM/FGF-1 chimera stimulated less SMC proliferation than did the wild-type FGF-1 with a median effective dose of approximately 0.3 nmol versus approximately 0.1 nmol (P <.001). By contrast, the chimera retained full stimulating activity on EC proliferation with a median effective dose of 0.06 nmol for both cytokines. Unlike the wild-type protein, the chimera possessed heparin-independent activity. In the absence of heparin, the chimera induced dose-dependent EC and SMC proliferation at 0.06 nmol or more compared with the wild-type FGF-1, which stimulated minimal DNA synthesis at 6.0-nmol concentrations. CONCLUSIONS: The HB-GAM/FGF-1 chimera displays significantly greater and uniquely heparin-independent mitogenic activity for both cell types, and in the presence of heparin it displays a significantly greater EC specificity.

The cysteine-free fibroblast growth factor 1 mutant induces heparin-independent proliferation of endothelial cells and smooth muscle cells.

BACKGROUND: The structure/function relationships of fibroblast growth factor 1 (FGF-1) are being investigated using site mutation, yielding novel structures with potential clinical applicability for modulating tissue responses to vascular interventions. We generated a mutant FGF-1 in which all three cysteines were converted to serines and then tested the relative mitogenic activities on endothelial cells (ECs) and smooth muscle cells (SMCs) and the molecular stability of the protein to thrombin-induced degradation. METHODS: The dose responses of wild-type FGF-1 and the Cys-free mutant in the absence or presence of heparin were tested on ECs and SMCs. Cell proliferation was measured by [(3)H]thymidine incorporation. Data were normalized by positive control (20% fetal bovine serum) and expressed as percentage of positive control for comparison. The molecular stability was examined by exposure of the cytokines to thrombin at 37 degrees C for 0.5-24 h and then analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. RESULTS: Unlike wild-type FGF-1 which induced only minimal DNA synthesis at concentrations as high as 100 ng/ml, the Cys-free mutant induced a dose-dependent proliferation starting at 1 ng/ml on both ECs and SMCs in the absence of heparin. At 100 ng/ml, Cys-free mutant induced 4-fold more proliferation than wild-type FGF-1 on ECs (76.64 +/- 13.39% vs 14.58 +/- 1.38%, P < 0.01) and 12-fold more proliferation on SMCs (143.52 +/- 9.96 vs 11.25 +/- 3.32, P < 0.01). Heparin 5 U/ml potentiated the mitogenic activity of the Cys-free mutant at low dose range. Both proteins were degraded by thrombin progressively. But the Cys-free mutant showed more susceptibility with accelerated appearance of lower-molecular-weight fragment bands after incubation with thrombin. CONCLUSIONS: Conversion of cysteine residues to serine changed the heparin dependency of the growth factor and increased its mitogenic activity and its susceptibility to thrombin-induced degradation.

Reporting the outcomes of care: effecting the transition from caring to comparing.

Accountability in health care has taken on new dimensions with the drive to base contracting and provider of care selection upon data driven "report cards." The measurement and development of reportable outcomes are driving providers to move their organizations from a focus not only on "caring" but to one highly cognizant of "comparing" in order to maintain market position and meet regulatory requirements. This article defines the areas of organizational transition required for reporting, and profiles the actions taken by three health care providers moving to an organizational style ready for "comparative" competition.

Modulation of vascular cell growth kinetics by local cytokine delivery from fibrin glue suspensions.

PURPOSE: Fibrin glue (FG) has been used as a delivery system for bioactive agents on grafts and angioplasty sites. Reports from two different institutions suggest that heparin concentrations of 500 U/mL in FG inhibit smooth muscle cell (SMC) proliferation, but do not effect endothelial cell (EC) proliferation. The purposes of this study were to (1) quantify the diffusive release of fibroblast growth factor-1 (FGF-1) and heparin from FG; (2) determine the effect of heparin and FGF-1 on SMC proliferation when the cells are immediately plated on the FG; and (3) by means of the diffusive release data, design a new in vitro model that may differentiate the effect of FG-incorporated FGF-1 and heparin, rather than the released, solubilized components of these two factors, on SMC and EC proliferation. METHODS: 125I-FGF-1 or 3H-heparin release from FG into the overlying media was measured serially in a 96-hour period, either with or without cells. SMCs were immediately plated on FG containing various concentrations of FGF-1 and heparin. SMCs or ECs were plated on identical groups of FG containing FGF-1 and heparin 24 hours after the FG was made to exclude the effect on cell growth of the initial release of FGF-1 into the media. RESULTS: In the first 24 hours, 70% +/- 1% of the FGF-1 and 59% +/- 2% of the heparin in the FG was released into the overlying media, with minimal release occurring thereafter. The cell type or absence of cells did not affect release, but there was five times more FGF-1 and four times more heparin in the media at 72 hours for the immediate plating versus the delayed plating because of a diffusive release primarily in the first 24 hours. A heparin concentration of 500 U/mL inhibited SMC proliferation, as compared with 5 U/mL heparin, only when immediate plating of SMCs was used. Comparing immediate versus delayed SMC plating, at equivalent FGF-1 and heparin doses, immediate plating induced greater proliferation than delayed plating; this was likely caused by the higher soluble FGF-1 concentration. Heparin doses as high as 500 U/mL had little effect on SMC proliferation. In contrast, ECs died with delayed plating on FG containing 500 U/mL heparin, and their growth was inhibited at 50 U/mL heparin, as compared with 5 U/mL heparin. CONCLUSION: The differences in SMC proliferation when comparing immediate versus delayed plating are likely caused by diffusive release of heparin and FGF-1 into the media. Our ongoing work uses an optimized in vitro FG system that minimizes the effects of soluble factors. This is an important distinction, because the cytokines that are released in vivo will be removed by blood flow and, thus, may not exert an effect unless they are contained within the FG.

S100A13 is involved in the regulation of fibroblast growth factor-1 and p40 synaptotagmin-1 release in vitro.

We have previously characterized the release of the signal peptide sequence-less fibroblast growth factor (FGF) prototype, FGF-1, in vitro as a stress-induced pathway in which FGF-1 is released as a latent homodimer with the p40 extravesicular domain of p65 synaptotagmin (Syn)-1. To determine the biologic relevance of the FGF-1 release pathway in vivo, we sought to resolve and characterize from ovine brain a purified fraction that contained both FGF-1 and p40 Syn-1 and report that the brain-derived FGF-1:p40 Syn-1 aggregate is associated with the calcium-binding protein, S100A13. Since S100A13 binds the anti-inflammatory compound amlexanox and FGF-1 is involved in inflammation, we examined the effects of amlexanox on the release of FGF-1 and p40 Syn-1 in response to stress in vitro. We report that while amlexanox was able to repress the heat shock-induced release of FGF-1 and p40 Syn-1 in a concentration-dependent manner, it had no effect on the constitutive release of p40 Syn-1 from p40 Syn-1 NIH 3T3 cell transfectants. These data suggest the following: (i) FGF-1 is associated with Syn-1 and S100A13 in vivo; (ii) S100A13 may be involved in the regulation of FGF-1 and p40 Syn-1 release in response to temperature stress in vitro; and (iii) the FGF-1 release pathway may be accessible to pharmacologic regulation.

Analysis of putative heparin-binding domains of fibroblast growth factor-1. Using site-directed mutagenesis and peptide analogues.

The contribution of individual basic amino acids within three putative "consensus sequences" for heparin binding of fibroblast growth factor-1 have been examined by site-directed mutagenesis. The results indicate that a significant reduction in the apparent affinity of fibroblast growth factor-1 for heparin is only observed when basic residues in one of the three regions are mutated. Mutation in the other regions are without affect on heparin binding. The heparin binding properties of synthetic peptides based on the three "consensus sequences" paralleled the mutagenesis results. That is, synthetic peptides corresponding to regions of the protein that were affected by mutagenesis with respect to heparin binding exhibited a relatively high affinity for immobilized heparin, whereas those corresponding to regions of similar charge density that were unaffected by mutagenesis did not. In addition, amino acid substitution of a nonbasic residue in the heparin-binding peptide could abolish its heparin binding capacity. The heparin-binding peptide could antagonize the mitogenic activity of FGF-1, probably because of the heparin dependence of this activity. Together these data demonstrate that the heparin binding properties of fibroblast growth factor-1 are dictated by structural features more complex than clusters of basic amino acids. The results of these and other studies indicate that consensus motifs for heparin-binding require further definition. More importantly, the results provide a basis for the design of peptide-based inhibitors of FGF-1.