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The mechanisms underlying ovarian dysfunction in polycystic ovary syndrome (PCOS) have not been definitively established. Our objective was to perform a detailed examination of ovarian responses to recombinant follicle-stimulating hormone (rFSH) in women with PCOS and controls.This prospective, crossover, dose-response study included three rFSH stimulation periods. Each stimulation period involved three consecutive, daily, subcutaneous injections of rFSH administered at a single dose. Low, medium and high rFSH doses were weight-adjusted, corresponding to 0.5, 1.1 and 2.2 IU/kg/d, respectively. Stimulation periods occurred in randomized order and were separated by 8-week washouts.Thirty participants (8 PCOS and 22 controls) were studied. PCOS was defined by oligomenorrhea and clinical or biochemical androgen excess, excluding other aetiologies of ovulatory dysfunction.Blood samples were obtained for hormone measurements before and 24 h after each rFSH injection.Participants with PCOS had significantly greater body mass index, antral follicle count and circulating testosterone, anti-mullerian hormone (AMH) and luteinizing hormone concentrations compared with controls participants. Baseline estradiol (E2) concentrations were similar in both groups. At the lowest dose of rFSH, PCOS participants did not demonstrate E2 increments, whereas a significant increase occurred in controls. rFSH-induced E2 production per follicle was significantly reduced in PCOS participants compared with controls at all rFSH doses. Increasing T and decreasing AMH concentrations were associated with augmented E2 production per follicle.Women with PCOS exhibited diminished initial E2 responses to rFSH compared with controls. These findings suggest that the mechanism of anovulation in PCOS may involve altered ovarian response to gonadotropins.

The Omicron BA.1 variant of SARS-CoV-2 escapes convalescent sera and monoclonal antibodies that are effective against earlier strains of the virus. This immune evasion is largely a consequence of mutations in the BA.1 receptor binding domain (RBD), the major antigenic target of SARS-CoV-2. Previous studies have identified several key RBD mutations leading to escape from most antibodies. However, little is known about how these escape mutations interact with each other and with other mutations in the RBD. Here, we systematically map these interactions by measuring the binding affinity of all possible combinations of these 15 RBD mutations (215 = 32,768 genotypes) to four monoclonal antibodies (LY-CoV016, LY-CoV555, REGN10987, and S309) with distinct epitopes. We find that BA.1 can lose affinity to diverse antibodies by acquiring a few large-effect mutations and can reduce affinity to others through several small-effect mutations. However, our results also reveal alternative pathways to antibody escape that do not include every large-effect mutation. Moreover, epistatic interactions are shown to constrain affinity decline in S309 but only modestly shape the affinity landscapes of other antibodies. Together with previous work on the ACE2 affinity landscape, our results suggest that escape of each antibody is mediated by distinct groups of mutations, whose deleterious effects on ACE2 affinity are compensated by another distinct group of mutations (most notably Q498R and N501Y).

Context Hyperandrogenism is a central feature of polycystic ovary syndrome (PCOS). In vitro studies have demonstrated that inflammatory stimuli promote whereas ibuprofen inhibits androgen production by ovarian theca-interstitial cells. Objective This work aimed to determine the effects of nonselective inhibitor of cyclooxygenases COX-1 and COX-2 on testosterone levels. Methods A prospective pilot study took place in an academic hospital of women with PCOS defined according to Rotterdam criteria (N = 20). Evaluations were taken at baseline and after 3 weeks of ibuprofen administration (400 mg twice a day or 400 mg 3 times a day, respectively, in women with weight Results Ibuprofen administration was associated with a decline of total testosterone from 0.75 ± 0.06 ng/mL to 0.59 ± 0.05 ng/mL (P = .008). There was no statistically significant change in the levels of other relevant hormones including dehydroepiandrosterone sulfate, gonadotropins, and insulin. Multiple regression analysis identified the greatest decline of testosterone was independently predicted by baseline testosterone level (P = .004) and by baseline insulin sensitivity index (P = .03). Conclusion Nonselective inhibition of COX-1 and COX-2 leads to selective reduction of testosterone consistent with direct inhibitory effect on ovarian steroidogenesis.

STUDY QUESTION Is polycystic ovary syndrome (PCOS) associated with an elevation of markers of endotoxemia? SUMMARY ANSWER In women with PCOS serum levels of lipopolysaccharides (LPS), the LPS to high-density lipoprotein (HDL) ratio and LPS-binding protein (LBP) are significantly greater than those of normal control subjects. WHAT IS KNOWN ALREADY Mononuclear cells from women with PCOS respond excessively to LPS by releasing pro-inflammatory cytokines. In rat ovarian theca-interstitial cell cultures LPS stimulates androgen production. STUDY DESIGN, SIZE, DURATION Cross-sectional study comparing markers of endotoxemia in women with PCOS (n = 62), healthy ovulatory women with polycystic ovary morphology (PCOM, n = 39) and a control group of healthy ovulatory women without PCOM [normal (NL), n = 43]. PARTICIPANTS/MATERIALS, SETTING, METHODS LPS was measured using a chromogenic assay. LBP was measured by ELISA. Total cholesterol and lipids were measured using a homogeneous enzyme colorimetric method. Androgens, gonadotrophins, prolactin, insulin, high-sensitivity C-reactive protein (hs-CRP) and sex hormone-binding globulin were determined by electrochemiluminescence assays. Glucose was measured using an enzymatic reference method with hexokinase. MAIN RESULTS AND THE ROLE OF CHANCE Women with PCOS, when compared with NL subjects, had a significantly higher mean LPS (P = 0.045), LPS/HDL ratio (P = 0.007) and LBP (P = 0.01). Women with PCOM had intermediate levels of markers of endotoxemia. Comparison among all groups revealed that markers of endotoxemia correlated positively with testosterone level, ovarian volume, number of antral follicles and hirsutism score, but negatively with the number of spontaneous menses per year. In multiple regression analysis, all measures of endotoxemia correlated independently and positively with hs-CRP and with ovarian volume. LIMITATIONS, REASONS FOR CAUTION This cross-sectional study reveals that markers of endotoxemia are associated with several clinical features observed in women with PCOS. However, responsible mechanisms and causation remain unknown. Steroid quantification was carried out by electrochemiluminescence assays and not by the current gold standard: liquid chromatography-mass spectrometry. Hence, the relationship of endotoxemia with features of PCOS and the extent to which endotoxemia contributes to reproductive and metabolic dysfunction warrants further investigation. WIDER IMPLICATIONS OF THE FINDINGS This study reveals the novel observation that markers of endotoxemia are elevated in young and otherwise healthy women with PCOS without significant metabolic dysfunction. Moreover, the association of clinical and endocrine markers of PCOS with those of endotoxemia may represent a pathophysiologic link to reproductive dysfunction as well as metabolic and long-term cardiovascular risks associated with this disorder. STUDY FUNDING/COMPETING INTEREST(S) Intramural funding from Poznan University of Medical Sciences. The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER N/A

Breast cancer is a leading cause of global cancer-related mortality. About 75% of breast tumors are estrogen receptor-positive (ER+). Nearly 30-50% of ER+ breast cancers eventually become refractory to multiple lines of therapy and progress to an uncurable metastatic stage despite positive 5-year results on primary endocrine therapy. To uncover the malignant phenotypes underlying the development of therapeutic resistance, we performed single-cell RNA sequencing (scRNA-seq) of 219,532 cells from 80 ER+ breast cancer patients. This included 28 solid tumors and 52 metastatic fluid samples (pleural effusions and ascites). We grouped samples in three groups to study the evolution of biological phenotypes based on the number of lines of therapy administered to a patient: 1. Early (non-metastatic), 2. Advanced (metastatic with less than 3 lines of therapy), and Refractory (metastatic with 3 or more lines of therapy). We used Pareto task inference analysis to model evolutionary task trade-offs across malignant cell populations from the early, advanced, and refractory tumors. Our model revealed 5 unique phenotypic states or “archetypes” that existed across ER+ cancer cells. A multitask learning model based on group lasso revealed the specialized biological tasks associated with each archetype, including 1. Metabolism and self-renewal, 2. EGF/STAT3 signaling, 3. Estrogen signaling, 4. Apoptosis, and 5. MYC activation with rapid proliferation. We found the estrogen signaling archetype was progressively lost in advanced and refractory tumors (P=7.2x10-110), while the MYC archetype was progressively gained in the advanced and refractory tumors (P=2.45x10-9). The EGF/STAT3 archetype was unique to refractory tumors (P= 1.42x10-37). We further investigated the evolution of resistant traits by modeling the pathway phenotypes in malignant cells against the number of lines of therapy received by the patients. Patients refractive to multiple lines of therapy progressively gained mTOR signaling pathway activity concurrent with loss of estrogen-signaling. Multiple pathways that crosstalk with the mTOR pathway were activated in the tumors receiving multiple lines of therapy, including IGFR, and EGF/ERBB signaling. In comparison to early and advanced tumors, refractory tumors also gained JNK/p38 signaling (P=0.006) and activated glycolysis (P=5x10-6). This study reveals how metastatic ER+ breast cancer cells acquire capabilities for resistance to multiple lines of therapy, thus serving as a unique resource for understanding refractory ER+ breast cancer biology. Citation Format: Aritro Nath, Patrick Cosgrove, Jason Griffiths, Jeffrey Chang, Adam Cohen, Andrea Bild. Single-cell RNA sequencing of early-stage to refractory metastatic ER+ breast cancers reveal phenotypic states underlying resistance to multiple lines of therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3129.

Over the past two decades, several broadly neutralizing antibodies (bnAbs) that confer protection against diverse influenza strains have been isolated1,2. Structural and biochemical characterization of these bnAbs has provided molecular insight into how they bind distinct antigens1. However, our understanding of the evolutionary pathways leading to bnAbs, and thus how best to elicit them, remains limited. Here, we measure equilibrium dissociation constants of combinatorially complete mutational libraries for two naturally isolated influenza bnAbs3–5 (CR-9114, 16 mutations; CR-6261, 11 mutations), reconstructing all possible intermediates back to the unmutated germline sequences. We find that these two libraries exhibit strikingly different patterns of breadth: while many variants of CR-6261 display moderate affinity to diverse antigens, those of CR-9114 display appreciable affinity only in specific, nested combinations. By examining the extensive pairwise and higher-order epistasis between mutations, we find key sites with strong synergistic interactions that are highly similar across antigens for CR-6261 and different for CR-9114. Together, these features of the binding affinity landscapes strongly favor sequential acquisition of affinity to diverse antigens for CR-9114, while the acquisition of breadth to more similar antigens for CR-6261 is less constrained. These results, if generalizable to other bnAbs, may explain the molecular basis for the widespread observation that sequential exposure favors greater breadth6–8, and such mechanistic insight will be essential for predicting and eliciting broadly protective immune responses.

Objectives In implementing the 2021 CKD-EPI without race equation in our institution, we sought to investigate the differences between the new CKD-EPI equation and the current MDRD equation to assist clinicians in transitioning to the new equation. Methods A retrospective study was performed in patients with eGFR Results eGFRs calculated by the 2021 CKD-EPI without race equation were well correlated to eGFRs calculated by the MDRD. However, notable differences were observed. Compared to the MDRD, the CKD-EPI w/o race equation for Black males with a serum creatinine less than 1.5 yielded decreased eGFR results with mean negative differences of -4.49 (18-60 years old), -6.43 (60-80 years old), and -8.87 (>80 years old). For serum creatinine cohorts between 1.5 to 2.0 and greater than 2.0, the differences were -3.35, -5.46, -7.48 and -1.86, -3.40, -4.14, respectively. A similar trend was noted in Black females. For those with a serum creatinine less than 1.5, the CKD-EPI equation yielded mean negative differences that increased with advancing age: -2.56, -4.74 and -7.27. For serum creatinine between 1.5 to 2.0 and greater than 2.0, the negative differences were -2.31, -3.73 and -5.28 and -1.30, -2.06 and -2.70, respectively. Therefore, in Black patients, the mean eGFR difference between the two equations increased as age increased and serum creatinine level decreased. In contrast, the CKD-EPI equation for White males with a serum creatinine less than 1.5 yielded increased eGFR results with mean positive difference of 7.22 (18-60 years old), 4.09 (60-80 years old), and 1.26 (>80 years old). For serum creatinine cohorts between 1.5 to 2.0 and greater than 2.0, the differences were 5.15, 2.54 and 0.18 and 2.01, 0.69 and -0.15, respectively. Similarly, White females had mean positive differences of 8.09 (18-60 years old), 4.61 (60-80 years old) and 2.00 (>80 years old). For cohorts with serum creatinine between 1.5 to 2.0 and greater than 2.0, the positive differences were 4.21, 2.24, 0.51 and 2.02, 0.82, 0.10. Therefore, in White patients, the difference of eGFR between the two equations increased as age decreased and creatinine level decreased. Conclusions Although the 2021 CKD-EPI equation strongly correlated with the MDRD, distinct differences exist. In our study, Black males and females exhibited a negative difference of around -5.0 and -4.0, respectively. The difference in eGFR was greater in those of older age and serum creatinine less than 1.5. Meanwhile, White males and females exhibited a positive difference of around 2.5 and 2.7. The difference in eGFR was greater in those of younger age and serum creatinine less than 1.5.

Background In the ovarian follicle, the Theca Cells (TCs) have two main functions: preserving morphological integrity and, importantly, secreting steroid androgen hormones. TCs express the essential enzyme 17α-hydroxylase/17,20-desmolase (CYP17), which permits the conversion of pregnenolone and progesterone into androgens. Dysregulation of CYP17 enzyme activity due to an intrinsic ovarian defect is hypothesized to be a cause of hyperandrogenism in women. Androgen excess is observed in women with polycystic ovary syndrome (PCOS) resulting from excess endogenous androgen production, and in transgender males undergoing exogenous testosterone therapy after female sex assignment at birth. However, the molecular and morphological effects of Cyp17 overexpression and androgen excess on folliculogenesis is unknown. Methods In this work, seeking a comprehensive profiling of the local outcomes of the androgen excess in the ovary, we generated a transgenic mouse model (TC17) with doxycycline (Dox)-induced Cyp17 overexpression in a local and temporal manner. TC17 mice were obtained by a combination of the Tet-dependent expression system and the Cre/LoxP gene control system. Results Ovaries of Dox-treated TC17 mice overexpressed Cyp17 specifically in TCs, inducing high testosterone levels. Surprisingly, TC17 ovarian morphology resembled the human ovarian features of testosterone-treated transgender men (partially impaired folliculogenesis, hypertrophic or luteinized stromal cells, atretic follicles, and collapsed clusters). We additionally assessed TC17 fertility denoting a perturbation of the normal reproductive functions (e.g., low pregnancy rate and numbers of pups per litter). Finally, RNAseq analysis permitted us to identify dysregulated genes (Lhcgr, Fshr, Runx1) and pathways (Extra Cellular Matrix and Steroid Synthesis). Conclusions Our novel mouse model is a versatile tool to provide innovative insights into study the effects of Cyp17 overexpression and hyperandrogenism in the ovary.

Polycystic ovary syndrome (PCOS) is a reproductive-metabolic disorder that is the most common endocrinopathy of women in their reproductive years of life. The reproductive phenotype includes hirsutism, irregular menstrual bleeding, and polycystic ovaries. Because of chronic anovulation, infertility is a major concern. Commonly, these features are accompanied by obesity, hyperlipidaemia, insulin resistance, and an increased risk for type 2 diabetes mellitus. In addition, there is increased prevalence of depression and anxiety disorders, acne, acanthosis nigricans, sleep apnoea, and metabolic syndrome. Genetic markers for PCOS have not been identified. The pathophysiology has been well described, but the underling basis for PCOS is unknown. The onset of symptoms occurs during or soon after puberty and there is a familial inheritance pattern. Long-term health concerns include an increased risk for uterine and cancer cardiovascular disease.

Objective To study the effects of ibuprofen on androgen production, gene expression, and cell viability in rat theca–interstitial cells exposed to the proinflammatory stimuli interleukin-1β (IL-1β) and lipopolysaccharide (LPS). Design Animal study. Setting University-based research laboratory. Patient(s)/Animal(s) Theca-interstitial cells were isolated from 30 day old female Sprague Dawley rats. Intervention(s) Theca cells were cultured with pro-inflammatory media containing IL-1β and LPS and compared with cells cultured in control media. Main Outcome Measure(s) Androstenedione quantification was performed on conditioned cell culture medium using liquid chromatography-mass spectrometry. Theca cell viability was assessed using PrestoBlue cell viability assay. The gene expression of Cyp17a1, Cyp11a1, and Hsd3b was analyzed using quantitative polymerase chain reaction. Result(s) Both proinflammatory stimuli IL-1β and LPS increased androstenedione concentration in cell culture medium, and these effects were mitigated with ibuprofen. Both inflammatory agents in addition increased the expression of key genes involved in androgen synthesis: Cyp17a1, Cyp11a1, and Hsd3b; the addition of ibuprofen to the culture medium inhibited these effects. Theca cell viability increased with IL-1β and LPS. Ibuprofen inhibited the IL-1β-mediated increase in cell viability but did not reverse the effects of LPS. Conclusion(s) In conclusion, our findings support the hypothesis that many of the alterations induced by inflammatory stimuli in theca–interstitial cells are abrogated by the addition of ibuprofen.

In a prospective, multicentre, randomised controlled trial in an average colon cancer risk population, the TOPAZ trial, Dr Cash and colleagues1 report that colon capsule endoscopy (CCE) revealed higher diagnostic yield (defined by finding a colonic polyp ≥6 mm that warrants an optical colonoscopy) and higher sensitivity compared with CT colonography (CTC). Based on these findings, Cash et al 1 advocate the use of CCE as a more accurate/superior diagnostic test among patients who cannot/prefer not to undergo optical colonoscopy. As abdominal radiologists experienced in CTC and as members of the American College of Radiology’s Colon Cancer Committee, we appreciate the effort by the authors in this clinical trial, but we have comments and concerns about the results, discussion and drawn conclusions

David J Ramsey,1,* Samuel J Poulin,1,* Lauren C LaMonica,2 Gregory R Blaha,1 Fina C Barouch,1 Jeffrey Chang,1 Jeffrey L Marx1 1Division of Ophthalmology, Lahey Hospital & Medical Center, Tufts University School of Medicine, Peabody, MA 01960, USA; 2University of Michigan Medical School, Ann Arbor, MI 48109, USA*These authors contributed equally to this workCorrespondence: David J RamseyLahey Hospital & Medical Center, Tufts University School of Medicine, 1 Essex Center Drive, Peabody, MA 01960, USAEmail David.J.Ramsey@lahey.orgPurpose: To evaluate the functional and anatomic outcomes, as well as cost-effectiveness, of the timing of conversion to intravitreal aflibercept (IVA) in patients with treatment-resistant diabetic macular edema (DME).Methods: Thirty consecutive eyes (25 patients) were identified that were treated with ≥ 3 intravitreal bevacizumab (IVB) and/or ranibizumab (IVR) injections prior to treatment with ≥ 3 IVA injections. Eyes that received ≤ 6 IVB and/or IVR injections (early-switch) were compared to those that received ≥ 7 injections (late-switch) prior to conversion to IVA. Treatment effectiveness was measured in quality-adjusted life years (QALYs). A micro-simulation model examined the impact of treatment duration on outcomes.Results: Early- (n=18) and late- (n=12) switch eyes had similar vision prior to conversion to IVA. Despite improvements in retinal thickness, only the early-switch eyes maintained vision gains after conversion to IVA through the end of follow-up (p=0.027). Early switch saved $22,884/eye and produced an additional 0.027 QALYs.Conclusion: Early conversion to IVA optimizes vision outcomes and results in lower overall treatment expenditures.Keywords: diabetic macular edema; DME, socioeconomics, diabetic retinopathy

Paracrine interactions between ovarian theca-interstitial cells (TICs) and granulosa cells (GCs) play an important role in the regulation of follicular steroidogenesis. Androgens serve as substrates for aromatization as well as affect GC function. This study evaluated the effects of co-culture of GC with TICs and the role of testosterone (T) and 5-alpha-dihydrotestosterone (DHT), and estradiol (E2) in modulation of GC expression of genes involved in the production of progesterone: 3β-hydroxysteroid dehydrogenase/Δ(5–4) isomerase (Hsd3b) and cholesterol side-chain cleavage (Cyp11). GCs obtained from immature Sprague-Dawley rats and were cultured in chemically defined media without or with TICs, DHT, or T. Hsd3b and Cyp11 transcripts were analyzed by qt-PCR. Co-culture of GCs with TICs stimulated Hsd3b and CYP11 expression in GCs. DHT and T induced a concentration-dependent upregulation of Hsd3b and CYP11 expression, as well as increased progesterone concentrations in spent media. E2 also increased expression of Hsd3b, and Cyp11. Effects of androgens were abrogated in the presence of an anti-androgen bicalutamide and the antiestrogen ICI 182780 (ICI). In conclusion, present findings demonstrate that androgens upregulate production of progesterone in GCs; these effects are likely due to a combination of direct action on androgen receptors and effects mediated by estrogen receptors.

Once unimaginable, fertility management is now a nationally established part of cancer care in institutions, from academic centers to community hospitals to private practices. Over the last two decades, advances in medicine and reproductive science have made it possible for men, women and children to be connected with an oncofertility specialist or offered fertility preservation soon after a cancer diagnosis. The Oncofertility Consortium's National Physicians Cooperative is a large-scale effort to engage physicians across disciplines – oncology, urology, obstetrics and gynecology, reproductive endocrinology, and behavioral health – in clinical and research activities to enable significant progress in providing fertility preservation options to children and adults. Here, we review the structure and function of the National Physicians Cooperative and identify next steps.

Objective Lower body fat is associated with diminishing cardiometabolic risk. Physiological differences between gluteofemoral and abdominal subcutaneous adipocyte functions are known, but the molecular basis for depot differences in adipocyte function is poorly understood. The objective of this study was to identify depot differences in microRNA (miRNA) expression in human abdominal and gluteofemoral subcutaneous adipose tissues and their implication in gene regulation. Methods Abdominal and gluteofemoral adipose tissue aspirates obtained from 18 participants (9 male and 9 female, age 30 ± 1.5 y, BMI 27.3 ± 1.23 kg/m2) were analyzed for miRNA expression profiles by next-generation DNA sequencing. The raw reads were mapped to miRBase 17, and differentially expressed miRNAs were confirmed by qRT-PCR. The hsa-mimic-miR196a was transfected into cultured abdominal preadipocytes isolated from five women with obesity. Target gene expression was evaluated by RT-qPCR. Results Among the 640 miRNAs detected in adipose tissue, miR196a2, miR196a1, miR196b, and miR204 showed a higher expression in the gluteofemoral depot (fold change = 2.7, 2.3, 1.7, and 2.3, respectively) independent of sex. Bioinformatic analyses and human primary preadipocyte transfection with miR196 suggested that the differentially expressed miRNAs could directly or indirectly modulate homeobox (HOX) gene expression. Conclusions The miR196 gene family could play an important role in the regulation of HOX gene expression in subcutaneous adipose tissue and in fat distribution variation.

Context: Peripubertal hyperandrogenemia—a precursor to polycystic ovary syndrome—is prominent in girls with obesity. Objective: We examined sources of overnight testosterone (T) and progesterone (P4) and potential sources of obesity-associated hyperandrogenemia during puberty. Design: Cross-sectional. Setting: Research unit. Participants/Interventions: Fifty girls ages 7 to 18 years—both normal weight (NW) and overweight (OW)—underwent dexamethasone (DEX) suppression (1 mg orally; 10 pm) and cosyntropin stimulation testing (0.25 mg intravenously; 8 am the following day), with blood sampled before and 30 and 60 minutes after cosyntropin. Thirty-nine subjects receiving DEX had frequent blood sampling overnight (every 10 minutes from 10 pm to 7 am) and were compared with 70 historical controls who did not receive DEX. Outcomes: Random coefficient regression modeling assessed changes in hormone concentrations after DEX and cosyntropin. Results: NW early pubertal controls exhibited early morning increases in free T and P4 levels; NW and OW late pubertal controls exhibited early morning increases in P4. Such changes were not observed in subjects receiving DEX. Post-DEX morning free T levels were fourfold higher in OW vs NW late pubertal girls. Postcosyntropin changes in free T and androstenedione were both 2.3-fold higher in OW vs NW late pubertal girls. Conclusions: These data suggest that (1) overnight increases in free T and P4 concentrations in NW early pubertal girls and P4 concentrations in late pubertal girls are of adrenal origin and (2) OW late pubertal girls demonstrate elevated nonadrenal free T levels after DEX and exaggerated adrenal androgen (free T and androstenedione) responses to cosyntropin., Overweight late pubertal girls demonstrated higher free testosterone (T) after dexamethasone suppression and greater free T and androstenedione responses to cosyntropin stimulation.

Infection with the intestinal helminth parasite Heligmosomoides polygyrus exacerbates the colitis caused by the bacterial enteropathogen Citrobacter rodentium. To clarify the underlying mechanism, we analyzed fecal microbiota composition of control and helminth-infected mice and evaluated the functional role of compositional differences by microbiota transplantation experiments. Our results showed that infection of Balb/c mice with H. polygyrus resulted in significant changes in the composition of the gut microbiota, characterized by a marked increase in the abundance of Bacteroidetes and decreases in Firmicutes and Lactobacillales. Recipients of the gut microbiota from helminth-infected wide-type, but not STAT 6-deficient, Balb/c donors had increased fecal pathogen shedding and significant worsening of Citrobacter-induced colitis compared to recipients of microbiota from control donors. Recipients of helminth-altered microbiota also displayed increased regulatory T cells and IL-10 expression. Depletion of CD4+CD25+ T cells and neutralization of IL-10 in recipients of helminth-altered microbiota led to reduced stool C. rodentium numbers and attenuated colitis. These results indicate that alteration of the gut microbiota is a significant contributor to the H. polygyrus-induced exacerbation of C. rodentium colitis. The helminth-induced alteration of the microbiota is Th2-dependent and acts by promoting regulatory T cells that suppress protective responses to bacterial enteropathogens.

Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder characterized by theca cell hyperplasia and excessive androgen production. An increasing body of evidence has pointed to a close association between PCOS and low-grade chronic systemic inflammation. However, the mechanistic basis for this linkage is unknown. Therefore, we evaluated the effects of the inflammatory agents lipopolysaccharide (LPS) and IL-1β on rat theca-interstitial cells (TICs). We found that incubation with either LPS or IL-1β elicited a dose-dependent increase in both TIC viability and androgen production. Using RNA sequencing analysis, we found that both of these inflammatory agents also triggered profound and widespread shifts in gene expression. Using a stringent statistical cutoff, LPS and IL-1β elicited differential expression of 5201 and 5953 genes, respectively. Among the genes upregulated by both LPS and IL-1β were key regulatory genes involved in the cholesterol and androgen biosynthesis pathways, including Cyp17a1, Cyp11a1, Hsd3b, and Hmgcr. This provides a molecular explanation for the mechanism of action of inflammatory agents leading to increased androgen production. Gene ontology and pathway analysis revealed that both LPS and IL-1β regulated genes highly enriched for many common functions, including the immune response and apoptosis. However, a large number of genes (n = 2222) were also uniquely regulated by LPS and IL-1β, indicating that these inflammatory mediators have substantial differences in their mechanism of action. Together, these findings highlight the potential molecular mechanisms through which chronic low-grade inflammation contributes to the pathogenesis of androgen excess in PCOS.

Double-bond photoisomerization in molecules such as the green fluorescent protein (GFP) chromophore can occur either via a volume-demanding one-bond-flip pathway or via a volume-conserving hula-twist pathway. Understanding the factors that determine the pathway of photoisomerization would inform the rational design of photoswitchable GFPs as improved tools for super-resolution microscopy. In this communication, we reveal the photoisomerization pathway of a photoswitchable GFP, rsEGFP2, by solving crystal structures of cis and trans rsEGFP2 containing a monochlorinated chromophore. The position of the chlorine substituent in the trans state breaks the symmetry of the phenolate ring of the chromophore and allows us to distinguish the two pathways. Surprisingly, we find that the pathway depends on the arrangement of protein monomers within the crystal lattice: in a looser packing, the one-bond-flip occurs, whereas in a tighter packing (7% smaller unit cell size), the hula-twist occurs.

Crohn’s disease (CD) is a multifactorial, chronic immune-mediated disorder. The oral cavity is involved in 0.5% to 20% of the patients with CD. Oral manifestations of CD are sometimes nonspecific and can be overlooked by the clinicians. These manifestations may precede intestinal symptoms and can serve as indicators for early diagnosis. To increase awareness and to contribute to the standard intervention, here we report a paediatric case with persistent idiopathic swelling of the lower lip and perianal fistula. Microscopic examinations revealed multiple non-necrotising granulomas with chronic inflammation, oedema and lymphangiectasia. The patient was treated with metronidazole 500 mg and ciprofloxacin 500 mg twice a day for one month. The perioral lesions were managed with topical 0.03% tacrolimus and oral prednisone 10 mgtwice a day for one month, followed by a tapered regimen of 10 mg/day for another month. The patient’s symptoms improved without full remission at the 6-month follow-up.

This paper represents an international collaboration of paediatric endocrine and other societies (listed in the Appendix) under the International Consortium of Paediatric Endocrinology (ICPE) aiming to improve worldwide care of adolescent girls with polycystic ovary syndrome (PCOS)1. The manuscript examines pathophysiology and guidelines for the diagnosis and management of PCOS during adolescence. The complex pathophysiology of PCOS involves the interaction of genetic and epigenetic changes, primary ovarian abnormalities, neuroendocrine alterations, and endocrine and metabolic modifiers such as anti-Müllerian hormone, hyperinsulinemia, insulin resistance, adiposity, and adiponectin levels. Appropriate diagnosis of adolescent PCOS should include adequate and careful evaluation of symptoms, such as hirsutism, severe acne, and menstrual irregularities 2 years beyond menarche, and elevated androgen levels. Polycystic ovarian morphology on ultrasound without hyperandrogenism or menstrual irregularities should not be used to diagnose adolescent PCOS. Hyperinsulinemia, insulin resistance, and obesity may be present in adolescents with PCOS, but are not considered to be diagnostic criteria. Treatment of adolescent PCOS should include lifestyle intervention, local therapies, and medications. Insulin sensitizers like metformin and oral contraceptive pills provide short-term benefits on PCOS symptoms. There are limited data on anti-androgens and combined therapies showing additive/synergistic actions for adolescents. Reproductive aspects and transition should be taken into account when managing adolescents.

Blood-based microarray studies comparing individuals affected with autism spectrum disorder (ASD) and typically developing individuals help characterize differences in circulating immune cell functions and offer potential biomarker signal. We sought to combine the subject-level data from previously published studies by mega-analysis to increase the statistical power. We identified studies that compared ex vivo blood or lymphocytes from ASD-affected individuals and unrelated comparison subjects using Affymetrix or Illumina array platforms. Raw microarray data and clinical meta-data were obtained from seven studies, totaling 626 affected and 447 comparison subjects. Microarray data were processed using uniform methods. Covariate-controlled mixed-effect linear models were used to identify gene transcripts and co-expression network modules that were significantly associated with diagnostic status. Permutation-based gene-set analysis was used to identify functionally related sets of genes that were over- and under-expressed among ASD samples. Our results were consistent with diminished interferon-, EGF-, PDGF-, PI3K-AKT-mTOR-, and RAS-MAPK-signaling cascades, and increased ribosomal translation and NK-cell related activity in ASD. We explored evidence for sex-differences in the ASD-related transcriptomic signature. We also demonstrated that machine-learning classifiers using blood transcriptome data perform with moderate accuracy when data are combined across studies. Comparing our results with those from blood-based studies of protein biomarkers (e.g., cytokines and trophic factors), we propose that ASD may feature decoupling between certain circulating signaling proteins (higher in ASD samples) and the transcriptional cascades which they typically elicit within circulating immune cells (lower in ASD samples). These findings provide insight into ASD-related transcriptional differences in circulating immune cells. © 2016 Wiley Periodicals, Inc.

Binding of mannose presenting macromolecules to the protein receptor concanavalin A (ConA) is investigated by means of single-molecule atomic force spectroscopy (SMFS) in combination with dynamic light scattering and molecular modeling. Oligomeric (Mw ≈ 1.5-2.5 kDa) and polymeric (Mw ≈ 22-30 kDa) glycomacromolecules with controlled number and positioning of mannose units along the scaffolds accessible by combining solid phase synthesis and thiol-ene coupling are used as model systems to assess the molecular mechanisms that contribute to multivalent ConA-mannose complexes. SMFS measurements show increasing dissociation force from monovalent (≈57 pN) to pentavalent oligomers (≈75 pN) suggesting subsite binding to ConA. Polymeric glycomacromolecules with larger hydrodynamic diameters compared to the binding site spacing of ConA exhibit larger dissociation forces (≈80 pN), indicating simultaneous dissociation from multiple ConA binding sites. Nevertheless, although simultaneous dissociation of multiple ligands could be expected for such multivalent systems, predominantly single dissociation events are observed. This is rationalized by strong coiling of the macromolecules' polyamide backbone due to intramolecular hydrogen bonding hindering unfolding of the coil. Therefore, this study shows that the design of glycopolymers for multivalent receptor binding and clustering must consider 3D structure and intramolecular interactions of the scaffold.

© 2015 by IGI Global. All rights reserved.Cloud computing refers to a scalable network infrastructure where consumers receive IT services such as software and data storage through the Internet on a subscription basis. Potential benefits include cost savings, simpler IT and reduced energy consumption. The UK government and local authorities, like commercial organisations, are considering cloud-based services. Concerns have been raised, however, over issues such as security, access, data protection and ownership. This study attempts to investigate the likely impact of cloud computing on local government based on a conceptual framework and case studies of four London borough councils. It reveals that the concept of cloud computing is new and not clearly understood. Local authorities, who face further cuts in government funding, welcome a cloud-based IT infrastructure which may lead to considerable savings. Yet local government is conservative, so with their risk-adverse attitude local authorities are more likely to adopt a hybrid approach to implementation.

Polycystic ovary syndrome is the most common endocrinopathy of reproductive-aged women. Major clinical manifestations include hirsutism and irregular menstrual bleeding as a result of altered hypothalamic-pituitary-ovarian interaction. Metabolic dysfunction is common and includes insulin resistance, lipid abnormalities, and obesity. There is increased risk for diabetes, uterine cancer, and cardiovascular disease.