Granulosa cell metabolism at ovulation correlates with oocyte competence and is disrupted by obesity and aging.

STUDY QUESTION: Is oocyte developmental competence associated with changes in granulosa cell (GC) metabolism? SUMMARY ANSWER: GC metabolism is regulated by the LH surge, altered by obesity and reproductive aging, and, in women, specific metabolic profiles are associated with failed fertilization versus increased blastocyst development. WHAT IS KNOWN ALREADY: The cellular environment in which an oocyte matures is critical to its future developmental competence. Metabolism is emerging as a potentially important factor; however, relative energy production profiles between GCs and cumulus cells and their use of differential substrates under normal in vivo ovulatory conditions are not well understood. STUDY DESIGN, SIZE, DURATION: This study identified metabolic and substrate utilization profiles within ovarian cells in response to the LH surge, using mouse models and GCs of women undergoing gonadotropin-induced oocyte aspiration followed by IVF/ICSI. PARTICIPANTS/MATERIALS, SETTING, METHODS: To comprehensively assess follicular energy metabolism, we used real-time metabolic analysis (Seahorse XFe96) to map energy metabolism dynamics (mitochondrial respiration, glycolysis, and fatty acid oxidation) in mouse GCs and cumulus-oocyte complexes (COCs) across a detailed time course in the lead up to ovulation. In parallel, the metabolic profile of GCs was measured in a cohort of 85 women undergoing IVF/ICSI (n = 21 with normal ovarian function; n = 64 with ovarian infertility) and correlated with clinical parameters and cycle outcomes. MAIN RESULTS AND THE ROLE OF CHANCE: Our study reveals dynamic changes in GC energy metabolism in response to ovulatory LH, with mitochondrial respiration and glycolysis differentially affected by obesity versus aging, in both mice and women. High respiration in GCs is associated with failed fertilization (P < 0.05) in a subset of women, while glycolytic reserve and mitochondrial ATP production are correlated with on-time development at Day 3 (P < 0.05) and blastocyst formation (P < 0.01) respectively. These data provide new insights into the cellular mechanisms of infertility, by uncovering significant associations between metabolism within the ovarian follicle and oocyte developmental competence. LIMITATIONS, REASONS FOR CAUTION: A larger prospective study is needed before the metabolic markers that were positively and negatively associated with oocyte quality can be used clinically to predict embryo outcomes. WIDER IMPLICATIONS OF THE FINDINGS: This study offers new insights into the importance of GC metabolism for subsequent embryonic development and highlights the potential for therapeutic strategies focused on optimizing mitochondrial metabolism to support embryonic development. STUDY FUNDING/COMPETING INTEREST(S): National Health and Medical Research Council (Australia). The authors have no competing interests. TRIAL REGISTRATION NUMBER: N/A.

Dynamic regulation of semaphorin 7A and adhesion receptors in ovarian follicle remodeling and ovulation.

The ovarian follicle is a complex structure that protects and helps in the maturation of the oocyte, and then releases it through the controlled molecular and structural remodeling process of ovulation. The progesterone receptor (PGR) has been shown to be essential in regulating ovulation-related gene expression changes. In this study, we found disrupted expression of the cellular adhesion receptor gene Sema7A in the granulosa cells of PGR-/- mice during ovulation. We subsequently found that expression of Sema7A in preovulatory follicles is promoted by gonadotropins and hypoxia, establishing an asymmetrical pattern with the SEMA7A protein enriched at the apex of large antral follicles. Sema7A expression was downregulated through a PGR-dependent mechanism in the periovulatory period, the abundance of SEMA7A protein was reduced, and the asymmetric pattern became more homogeneous after an ovulatory stimulus. Receptors for Sema7A can either repel or promote intercellular adhesion. During ovulation, striking inverse regulation of repulsive Plxnc1 and adhesive Itga5/Itgb1 receptors likely contributes to dramatic tissue remodeling. The adhesive receptor Itga5 was significantly increased in periovulatory granulosa cells and cumulus-oocyte complexes (COCs), and functional assays showed that periovulatory granulosa cells and COCs acquire increased adhesive phenotypes, while Sema7A repels granulosa cell contact. These findings suggest that the regulation of Sema7A and its associated receptors, along with the modulation of integrin α5, may be critical in establishing the multilaminar ovarian follicle structure and facilitating the remodeling and apical release of the cumulus-oocyte complex during ovulation.

Characterisation of saponins from Hedera nepalensis in Vietnam northwest mountainous areas with the aid of high-resolution mass spectrometry.

This research aims to explore the saponins composition of H. nepalensis in four northwest mountainous areas of Vietnam including Ha Giang, Lai Chau, Lao Cai, and Lang Son with the aid of high-resolution mass spectrometry. As a result, 42 saponins are successfully identified in H. nepalensis leaves by UHPLC-Q-TOF-MS/MS analyses, in which two 30-noroleanane and four oleanane triterpene saponins structures have been reported for the first time. Two structures of compound 20 were discovered in four samples. Two structures of compound 8 were found in H. nepalensis from Ha Giang and Lao Cai, while two structures of compound 28 were not observed in Lang Son. Different environmental and climatic circumstances in various places may have an impact on chemical constituents of H. nepalensis. By providing the phytochemicals profile of H. nepalensis leaves, our study supports the orientation for future research on this medicinal plant as well as other Hedera species.

Progesterone receptor mediates ovulatory transcription through RUNX transcription factor interactions and chromatin remodelling.

Progesterone receptor (PGR) plays diverse roles in reproductive tissues and thus coordinates mammalian fertility. In the ovary, rapid acute induction of PGR is the key determinant of ovulation through transcriptional control of a unique set of genes that culminates in follicle rupture. However, the molecular mechanisms for this specialized PGR function in ovulation is poorly understood. We have assembled a detailed genomic profile of PGR action through combined ATAC-seq, RNA-seq and ChIP-seq analysis in wildtype and isoform-specific PGR null mice. We demonstrate that stimulating ovulation rapidly reprograms chromatin accessibility in two-thirds of sites, correlating with altered gene expression. An ovary-specific PGR action involving interaction with RUNX transcription factors was observed with 70% of PGR-bound regions also bound by RUNX1. These transcriptional complexes direct PGR binding to proximal promoter regions. Additionally, direct PGR binding to the canonical NR3C motif enable chromatin accessibility. Together these PGR actions mediate induction of essential ovulatory genes. Our findings highlight a novel PGR transcriptional mechanism specific to ovulation, providing new targets for infertility treatments or new contraceptives that block ovulation.

Nuclear Receptors in Ovarian Function.

The ovary undergoes cycles of hormone production that regulate physiological changes necessary for folliculogenesis, ovulation and luteinisation, ultimately contributing to female reproductive success. Crucial to these biological processes is stage-specific nuclear receptor signalling. While the transcriptional regulatory roles of steroid receptors in female fertility and especially ovarian functions have long been documented, non-steroid receptors also play an important part in regulating gene expression at various stages of ovarian development. The recent application of high-throughput genomic and transcriptomic technologies has begun to shed light on the molecular mechanisms underlying ovarian nuclear receptor actions and pointed to a complex interplay between highly specific transcription co-regulators as well as between nuclear receptors in mediating mutual as well as unique target genes. Interrelationships between nuclear receptors as well as the involvement of context-specific protein and non-protein co-regulators are likely keys to the precise and specific nuclear receptor action in the ovary. Leveraging such knowledge on the nuclear receptor network is especially valuable in the development of novel fertility treatments as well as female contraceptives.

Reproductive biology research down under: highlights from the Australian and New Zealand Annual Meeting of the Society for Reproductive Biology, 2021.

Against the backdrop of a global pandemic, the Society for Reproductive Biology (SRB) 2021 meeting reunited the Australian and New Zealand reproductive research community for the first time since 2019 and was the first virtual SRB meeting. Despite the recent global research disruptions, the conference revealed significant advancements in reproductive research, the importance of which span human health, agriculture, and conservation. A core theme was novel technologies, including the use of medical microrobots for therapeutic and sperm delivery, diagnostic hyperspectral imaging, and hydrogel condoms with potential beyond contraception. The importance of challenging the contraceptive status quo was further highlighted with innovations in gene therapies, non-hormonal female contraceptives, epigenetic semen analysis, and in applying evolutionary theory to suppress pest population reproduction. How best to support pregnancies, particularly in the context of global trends of increasing maternal age, was also discussed, with several promising therapies for improved outcomes in assisted reproductive technology, pre-eclampsia, and pre-term birth prevention. The unique insights gained via non-model species was another key focus and presented research emphasised the importance of studying diverse systems to understand fundamental aspects of reproductive biology and evolution. Finally, the meeting highlighted how to effectively translate reproductive research into policy and industry practice.

Intraovarian, Isoform-Specific Transcriptional Roles of Progesterone Receptor in Ovulation.

Progesterone receptor (PGR) activity is obligatory for mammalian ovulation; however, there is no established direct functional pathway explaining how progesterone receptor completely and specifically regulates oocyte release. This study examined the overarching cell- and isoform-specific effects of the PGR within each cellular compartment of the ovary, using mice null for the PGR (PRKO), as well as isoform-specific null mice. The PGR was expressed in ovarian granulosa and stromal cells and although PRKO ovaries showed no visible histological changes in preovulatory ovarian morphology, follicle rupture did not occur. Reciprocal ovarian transplant experiments established the necessity of ovarian PGR expression for ovulation. Cumulus-oocyte complexes of PRKO mice exhibited normal morphology but showed some altered gene expression. The examination of mitochondrial activity showed subtle differences in PRKO oocytes but no differences in granulosa cell respiration, glycolysis or ß-oxidation. Concurrently, RNA-seq identified novel functional pathways through which the PGR may regulate ovulation. PGR-A was the predominant transcriptionally active isoform in granulosa cells and 154 key PGR-dependent genes were identified, including a secondary network of transcription factors. In addition, the PGR regulated unique gene networks in the ovarian stroma. Collectively, we establish the effector pathways activated by the PGR across the ovarian cell types and conclude that PGR coordinates gene expression in the cumulus, granulosa and stromal cells at ovulation. Identifying these networks linking the PGR to ovulation provides novel targets for fertility therapeutics and nonhormonal contraceptive development.

Sex-Specific Control of Human Heart Maturation by the Progesterone Receptor.

BACKGROUND: Despite in-depth knowledge of the molecular mechanisms controlling embryonic heart development, little is known about the signals governing postnatal maturation of the human heart. METHODS: Single-nucleus RNA sequencing of 54 140 nuclei from 9 human donors was used to profile transcriptional changes in diverse cardiac cell types during maturation from fetal stages to adulthood. Bulk RNA sequencing and the Assay for Transposase-Accessible Chromatin using sequencing were used to further validate transcriptional changes and to profile alterations in the chromatin accessibility landscape in purified cardiomyocyte nuclei from 21 human donors. Functional validation studies of sex steroids implicated in cardiac maturation were performed in human pluripotent stem cell-derived cardiac organoids and mice. RESULTS: Our data identify the progesterone receptor as a key mediator of sex-dependent transcriptional programs during cardiomyocyte maturation. Functional validation studies in human cardiac organoids and mice demonstrate that the progesterone receptor drives sex-specific metabolic programs and maturation of cardiac contractile properties. CONCLUSIONS: These data provide a blueprint for understanding human heart maturation in both sexes and reveal an important role for the progesterone receptor in human heart development.

Antiproliferative activity and apoptosis induction by trijuganone C isolated from the root of Salvia miltiorrhiza Bunge (Danshen).

In this study, we found that the hexane fraction of Danshen, the dried root of Salvia miltiorrhiza (Lamiaceae), exerted antiproliferative effects on human leukemia cells. Phytochemical investigation of the hexane fraction achieved the isolation of the tanshinone diterpenes: dihydrotanshinone I (1), trijuganone C (2), trijuganone B (3), cryptotanshinone (4), tanshinone IIA (5), and tanshinone I (6). Compound 2 showed significant antiproliferative activities against human leukemia cells HL-60, Jurkat, and U937. The antiproliferative activities of 2 against human cancer and normal cells indicated that 2 exhibited potent antiproliferative activities with IC50 values less than 10 µM against HL-60 and Jurkat cells as well as on the colon cancer cells DLD-1, COLO 205, and Caco-2. Compound 2 induced chromatin condensation, DNA fragmentation, activation of caspase-3, -8, and -9, and the cleavage of poly (ADP-ribose) polymerase (PARP) in HL-60 cells. Moreover, 2 activated Bid and Bax, leading to the loss of mitochondrial membrane potential, and 2 induced the cytochrome c release from mitochondria into cytosol. In contrast, Bcl-2 and Bcl-xL were unaffected by 2. These results suggest that 2 exerts antiproliferative effects via apoptosis induction mediated by mitochondrial dysfunction and caspase activation. Compound 2 may serve as a candidate of potential chemotherapeutic agent for human leukemia.

Bioactive Triterpenes from the Root of Salvia miltiorrhiza Bunge.

Danshen (Salvia miltiorrhiza) is a well-known medicinal herb in the oriental medicine. The current study on bioactive triterpenoid in the root of S. miltiorrhiza led to the isolation of a new highly hydroxylated ursane-type triterpene, urs-12-ene-2α,3ß,7ß,16α-tetraol (1) and five known ones including 2ß-hydroxypomolic acid (2), maslinic acid (3), asiatic acid (4), ursolic acid (5), and oleanolic acid (6). Their structures were elucidated on the basis of extensive spectroscopic analyses and comparison with literature data. The antiproliferative testing against HL-60 cells revealed that the new compound 1 and ursolic acid (5) showed weak and moderate activities with IC50 values of 42.2 and 11.7 µM. In addition, compounds 1-3 showed inhibitory effect on ghrelin activity. Copyright © 2017 John Wiley & Sons, Ltd.

Flow control using audio tones in resonant microfluidic networks: towards cell-phone controlled lab-on-a-chip devices.

Fluid control remains a challenge in development of portable lab-on-a-chip devices. Here, we show that microfluidic networks driven by single-frequency audio tones create resonant oscillating flow that is predicted by equivalent electrical circuit models. We fabricated microfluidic devices with fluidic resistors (R), inductors (L), and capacitors (C) to create RLC networks with band-pass resonance in the audible frequency range available on portable audio devices. Microfluidic devices were fabricated from laser-cut adhesive plastic, and a "buzzer" was glued to a diaphragm (capacitor) to integrate the actuator on the device. The AC flowrate magnitude was measured by imaging oscillation of bead tracers to allow direct comparison to the RLC circuit model across the frequency range. We present a systematic build-up from single-channel systems to multi-channel (3-channel) networks, and show that RLC circuit models predict complex frequency-dependent interactions within multi-channel networks. Finally, we show that adding flow rectifying valves to the network creates pumps that can be driven by amplified and non-amplified audio tones from common audio devices (iPod and iPhone). This work shows that RLC circuit models predict resonant flow responses in multi-channel fluidic networks as a step towards microfluidic devices controlled by audio tones.

Association between nutritional status and dengue infection: a systematic review and meta-analysis.

BACKGROUND: Dengue infection has various clinical manifestations, often with unpredictable clinical evolutions and outcomes. Several factors including nutritional status have been studied to find the relationship with dengue severity. However, the nutritional status had conflicting effects on the complication of dengue in some previous studies. Therefore, we conducted a systematic review and performed a meta-analysis to analyze the association between nutritional status and the outcome of dengue infection. METHODS: Eleven electronic databases and manual searching of reference lists were used to identify the relevant studies published before August 2013. At least two authors worked independently in every step to select eligible studies and extract data. Dengue severity in the included studies must be classified into three categories: dengue fever (DF), dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). RESULTS: Thirteen articles that met the inclusion criteria came to final analysis. A meta-analysis using fixed- or random-effects models was conducted to calculate pooled odds ratios (OR) with corresponding 95 % confidence intervals. It has shown that there was no statistically significant association between DHF group and DSS group in malnutritional and overweight/obesity patients with OR: 1.17 (95 % CI: 0.99-1.39), 1.31 (0.91-1.88), respectively. A significantly inverse relation between DF and DHF groups of malnutritional patients was revealed (OR = 0.71, 95 % CI: 0.56-0.90). Our meta-analysis also indicated a statistically significant negative correlation between malnourished children with dengue virus infection and healthy children (OR = 0.46, 95 % CI: 0.3-0.70). When analyzing patients with normal nutrition status, we found out that there was a significantly negative relationship between DHF and DSS groups (0.87; 95 % CI: 0.77-0.99). Other comparisons of DSS with DF/DHF groups, DSS/DHF with DF groups, and DHF with DF groups in normal nutritional patients showed no significant correlation. However, the findings should be interpreted cautiously because all significant associations were lost after removing of the largest study. CONCLUSIONS: Results from previous studies failed to show any solid consistency regarding the association between the nutritional status and dengue infection. Consequently, the effects of nutritional status on dengue disease outcome has been controversial. Further studies are recommended to clarify the impact of nutritional status on dengue infection.

[Surgery and curietherapy of keloids]. / Chirurgie et curiethérapie des chéloïdes.

Postoperative irradiation of keloids allows a decrease of the recurrence rate by about 50%, compared to surgery alone. A review of the literature illustrates the benefits due to the irradiation, and describes the techniques available. The Iridium 192 interstitial brachytherapy, with per-operative implantation of the plastic tubes and immediate irradiation of 20 Gy at 5 mm depth, is detailed as used by French teams. The analysis of the published results allows to recommend this technique which is tailored to each clinical situation, safe, and easy to perform by the surgeon.

[Temporomandibular joint luxation and Ehlers-Danlos disease. Apropos of a case]. / Luxation temporo-mandibulaire et maladie d'Ehlers-Danlos.

First described by Tschernogobow in 1981, Ehlers-Danlos syndrome is usually observed in white males. Symptoms results from defective collagen synthesis. Diagnosis is based on clinical presentation. There are 9 different clinical groups. Maxillofacial manifestations are usually seen in type VIII Ehlers-Danlos syndrome. The clinical case presented here illustrates the problems involving the temporomandibular joints and focuses on an assessment of proposed therapeutic options.

[Traumatic rupture of the levator tendon of the upper eyelid]. / La désinsertion traumatique du releveur de la paupière supérieure.

We report on two cases of traumatic desinsertion of levator aponeurosis. After a blunt trauma diagnosis between levator aponeurosis desinsertion and neurogenic ptosis is important in planing the treatment: early surgery for the first and foregoing for the later.

[The first- and second-arch syndrome. A treatment analysis and plan]. / Syndrome du premier et du deuxième arc. Analyse et plan de traitement.

There is a very great lesion staging in these syndromes, and a great number of procedures have been described, according to the age and severity of the lesions. The initial assessment is therefore important, since it allows classifying these malformations according to their severity in key sectors. Scheduling the treatment in time must take account of: growth, the necessity to perform successive operations in one region, since no operation must hinder a subsequent one. It is essential to determine the optimum time for bone reconstruction. An early treatment with a conventional bone graft, without any intrinsic growth potential, condemns the reconstructed region to immobility, therefore to a progressive degradation of the result in time and to successive corrections. Early reconstruction is justified only if the available reconstruction means allow the reconstructed area to grow, either naturally or with the aid of orthopaedic stimulation. Some means seem to be available to date. A choice must therefore be made between: delayed morphological surgery, early functional surgery. Considering the extent of the means implemented, this is reserved for severe cases. The other great problem is the reconstruction of the auricle, which most often requires a series of operations scheduled over 2 years. The middle ear may not be operated if the lesions are unilateral. The other malformations, including macrostomia, muscular and neural abnormalities, involvement of the eyelids, sometimes require correction, which must fit in a repair schedule that must be established as soon as possible and must take account of the predictable procedures in order to prevent them from hindering each other.

[Osteosarcomas of the mandible]. / Les ostéosarcomes de la mandibule.

On the base of 3 cases of mandibular osteosarcoma, the current role of surgery is specified. The most often used procedure is Rosen's protocol, which includes: preoperative chemotherapy, radical surgery with reconstruction, and postoperative chemotherapy. The key element is the pathological study of the surgical specimen. This will determine the response of the tumor to preoperative chemotherapy and, by comparing the histology of the initial biopsy specimen to that of the surgical specimen, will be used as a basis to select the postoperative chemotherapy. For this type of tumor, chemotherapy has completely substituted for radiation therapy, which is no longer used in the initial treatment. Prophylactic lung radiation therapy has also been given up due to the risks of pulmonary fibrosis entailed by the effective doses. Surgery still has a role of choice in the therapeutic schedule, even if it cannot be contemplated as the sole treatment. Considering the usually young age of the patients, we currently think that reconstruction must be immediate and must resort to all the techniques likely to produce the best possible functional and morphological result.

[Bone substitutes and infection in maxillofacial surgery]. / Substituts osseux et infection en chirurgie maxillo-faciale.

Bony substitutes are in fashion in maxillofacial surgery. They are used to fill bony cavities, in bony reconstruction to give shape-lines and re-create area of support or to fill bony defects. Then their use is frequently crowned by success in general surgery, it is not the same thing in maxillofacial surgery because of the usual impossibility to fulfil a requirement to biomaterial utilisation: the watertight. Furthermore, they don't have yet, for the most part, an essential quality: malleability. In our experience, their use are frequently disappointing because of postoperative infections and we stay faithful to the autograft bone. However it is highly probable that these biomaterials will take an importance more and more considerable when these problems will be overcame.

[A case of cellular blue nevus with bone and meningeal invasion]. / A propos d'un cas de naevus bleu cellulaire avec envahissement osseux et méningé.

The blue naevus is an essentially benign tumour. However, the cellular variety may rarely undergo malignant transformation. The authors report an unusual case of blue naevus in terms of size and evolution. This tumour which involved the occipitovertebral and parotid regions infiltrated underlying tissues and organs, in particular, the occipital bone, the dura mater and the parotid gland. The clinical, histological and histogenic problems associated with the blue naevus are discussed. There are only scattered reports of tumours of a similar nature in the literature and for this reason we felt that this case merited being reported.

[Value of MRI in the visualization of the meniscus in temporomandibular joints]. / L'intérêt de l'IRM dans la visualisation du ménisque des articulations temporo-mandibulaires (A.T.M.).

Modern imaging techniques, among which nuclear magnetic resonance currently ranks first, have made it much easier to understand the internal disorders of the temporomandibular joint. While computed tomography still is the ideal technique for all bony affections in the articular region, especially for injuries. MRI now is an essential first-intention complementary examination. It allows assessing the location of the meniscus and very well defines the pathology being explored: reducible or irreducible dislocation of the meniscus, either purely anterior or anterolateral, the latter being better visible on coronal views (which are not systematically taken). The examination includes T1-weighted parasagittal views perpendicular to the axis of the condyle, taken with a surface coil (knee coil). Two sequences are performed, one with the mouth open and one with the mouth shut. The meniscus appears as a biconcave hypointense signal, which normally lies on the head of the condyle both when the mouth is open and when it is shut. T2-weighted sequences have the advantage of revealing possible effusion or intra-articular adhesion between the capsule and the synovial membrane. They may also better demonstrate a possible myxoid degeneration of the meniscus. The considerable increase in scanning time required for these sequences accounts for their unfrequent use. The condition of the bone can also be assessed, but we find MRI less accurate than direct sagittal CT scans. The future prospects include shorter imaging times owing to fast-imaging sequences (short T1 with partial flip angle).