Pregnancy (im)possibilities: identifying factors that influence sexual minority women’s pregnancy desires

ABSTRACT

Sexual minority women (SMW) face both increased risk for unintended pregnancy and barriers to achieving wanted pregnancy, but little research investigates SMW’s pregnancy desires. To fill this gap, we conducted five focus groups and 11 in-depth interviews with 20-30-year-old SMW in three US cities. Findings highlight that the heteronormative pregnancy planning paradigm lacks salience for SMW. While some SMW clearly wish to avoid pregnancy, many others are unsure, and factors influencing this uncertainty include relationship context, anticipating logistical barriers, and discord between queer identity and pregnancy.     

New antimicrobial flavonoids and chalcone from <Emphasis Type="Italic">Colutea armata</Emphasis>

Colucins A (1) and B (2), new flavonoids and colucone (3), the new chalcone derivative, have been isolated from the CHCl₃-soluble fraction of the whole plant of Colutea armata along with luteolin (4), luteolin 7-O-β-D-glucoside (5), isoliquiritigenin (6), trans-caffeic acid (7) and stigmasterol (8) reported for the first time from this species. Their structures were elucidated by spectroscopic techniques including MS and 2D-NMR spectroscopy. Compounds 1 and 2 showed significant antimicrobial activity against two Gram positive and three Gram negative bacterial strains while 3 was moderately active.     

Understanding Factors Associated with Postpartum Visit Attendance and Contraception Choices: Listening to Low-Income Postpartum Women and Health Care Providers

Background While there is considerable variability with respect to attendance at the postpartum visit, not much is known about women’s preferences with respect to postpartum care. Likewise, there is also limited information on providers’ practices regarding the postpartum visit and care including the delivery of contraception. To understand and address deficits in the delivery and utilization of postpartum care, we examined the perceptions of low-income postpartum women with respect to barriers to and preferences for the timing and location of the postpartum visit and receipt of contraception. We also examined providers’ current prenatal and postnatal care practices for promoting the use of postpartum care and their attitudes toward alternative approaches for delivering contraceptive services in the postpartum period. Methods Qualitative face-to-face interviews were completed with 20 postpartum women and in-depth qualitative phone interviews were completed with 12 health care providers who had regular contact with postpartum women. Interviews were coded using Atlas.ti software and themes were identified. Results Women believed that receiving care during the postpartum period was an important resource for monitoring physical and mental health and also strongly supported the provision of contraception earlier than the 6-week postpartum visit. Providers reported barriers to women’s use of postpartum care on the patient, provider, and system levels. However, providers were receptive to exploring new clinical practices that may widen the reach of postpartum care and increase access to postpartum contraception. Conclusion Approaches that increase the flexibility and convenience of postpartum care and the delivery of postpartum contraception may increase the likelihood that women will take advantage of essential postpartum services.     

SPATA33 localizes calcineurin to the mitochondria and regulates sperm motility in mice

Calcineurin is a calcium-dependent phosphatase that plays roles in a variety of biological processes including immune responses. In spermatozoa, there is a testis-enriched calcineurin composed of PPP3CC and PPP3R2 (sperm calcineurin) that is essential for sperm motility and male fertility. Because sperm calcineurin has been proposed as a target for reversible male contraceptives, identifying proteins that interact with sperm calcineurin widens the choice for developing specific inhibitors. Here, by screening the calcineurin-interacting PxIxIT consensus motif in silico and analyzing the function of candidate proteins through the generation of gene-modified mice, we discovered that SPATA33 interacts with sperm calcineurin via a PQIIIT sequence. Spata33 knockout mice exhibit reduced sperm motility because of an inflexible midpiece, leading to impaired male fertility, which phenocopies Ppp3cc and Ppp3r2 knockout mice. Further analysis reveals that sperm calcineurin disappears from the mitochondria in the Spata33 knockout testis. In addition, immunoprecipitation analysis indicates that sperm calcineurin interacts with not only SPATA33 but also the mitochondrial protein VDAC2. These results indicate that SPATA33 localizes calcineurin to the mitochondria and regulates sperm motility.

Fertilization is the union of two gametes, spermatozoa and eggs. In the female reproductive tract, spermatozoa need to travel a long distance to reach the egg and pass through the zona pellucida (ZP), an extracellular matrix that surrounds the egg. Sperm motility plays critical roles in these processes, which is executed by the flagellum that can be divided into three parts: midpiece, principal piece, and end piece (1, 2). The central motility apparatus of the flagellum is the axoneme, a “9+2” microtubule structure, that is found in all the parts of the flagellum. In addition to the axoneme, the midpiece possesses spirally arranged mitochondria called the mitochondrial sheath. The principal piece possesses a fibrous sheath that provides elastic rigidity and a scaffold for glycolytic and signaling molecules (1). The end piece contains no accessory structures. Defects in these structures’ formation or function could lead to impaired sperm motility and male infertility (1, 2).Calcineurin is a calcium-dependent phosphatase that is evolutionarily conserved from yeasts to mammals and is comprised of two subunits, a catalytic and a regulatory subunit. In mammals, three isoforms (PPP3CA, PPP3CB, and PPP3CC) of the catalytic subunit and two isoforms (PPP3R1 and PPP3R2) of the regulatory subunit have been identified. Ppp3ca, Ppp3cb, and Ppp3r1 are expressed ubiquitously and play roles in a variety of biological processes, including immune responses and cardiac morphogenesis (3, 4). In contrast, PPP3CC and PPP3R2 compose the testis-enriched calcineurin (sperm calcineurin), the disruption of which leads to an inflexible midpiece, impaired sperm motility, and male infertility (5–7). The administration of calcineurin inhibitors such as cyclosporine A and tacrolimus (FK506), both widely used immunosuppressant drugs, to wild-type (WT) males phenocopied the Ppp3cc and Ppp3r2 knockout (KO) mice (5). These defects appear within 4 to 5 d of treatment, and male fertility recovered 1 wk after halting the drug administration, suggesting that sperm calcineurin can be a target for reversible and rapidly acting male contraceptives (5). However, it is challenging to develop molecules that specifically inhibit sperm calcineurin and not somatic calcineurin because of sequence similarities (82% amino acid identity between human PPP3CA and PPP3CC and 85% amino acid identity between human PPP3R1 and PPP3R2). Therefore, identifying proteins that interact with sperm calcineurin widens the choice of inhibitors that target the sperm calcineurin pathway.The PxIxIT motif is a conserved sequence found in calcineurin-binding proteins (8, 9). Proteins with this consensus motif can be a substrate of calcineurin. For example, NFAT, in which the PxIxIT motif was first discovered, is translocated to the nucleus and activates gene expression when it is dephosphorylated by calcineurin, which plays critical roles in immune responses (10). In addition, PxIxIT motif-containing proteins can be regulators of calcineurin. For example, RCAN1 is an endogenous inhibitor of calcineurin (11, 12), while AKAP5 anchors calcineurin to the L-type Ca²⁺ channel in the plasma membrane (13, 14). In contrast to a variety of molecules known to bind to somatic calcineurin, little is known about the proteins that interact with sperm calcineurin.Here, we searched the testis-enriched proteins that contain the PxIxIT consensus motif to identify substrates or regulators of sperm calcineurin, which may help not only reveal how sperm calcineurin works in regulating sperm motility but also discover new male contraceptive targets and understand reproductive toxicities that can be caused by calcineurin inhibitor–based immunosuppression. By analyzing the function of candidate proteins through the generation of gene-modified mice with the CRISPR/Cas9 system, we identified that SPATA33 is essential in localizing sperm calcineurin to the mitochondria and regulating sperm motility.     

Biological Transformation of Zearalenone by Some Bacterial Isolates Associated with Ruminant and Food Samples

Zearalenone (ZEA) is a secondary metabolite produced by Fusarium spp., the filamentous fungi. Food and feed contamination with zearalenone has adverse effects on health and economy. ZEA degradation through microorganisms is providing a promising preventive measure. The current study includes isolation of 47 bacterial strains from 100 different food and rumen samples. Seventeen isolates showed maximum activity of ZEA reduction. A bacterial isolate, RS-5, reduced ZEA concentration up to 78.3% through ELISA analysis and 74.3% as determined through HPLC. Ten of the most efficient strains were further selected for comparison of their biodegradation activity in different conditions such as incubation period, and different growth media. The samples were analyzed after 24 h, 48 h, and 72 h of incubation. De Man Rogosa Sharp (MRS) broth, Tryptic soy broth, and nutrient broth were used as different carbon sources for comparison of activity through ELISA. The mean degradation % ± SD through ELISA and HPLC were 70.77% ± 3.935 and 69.11% ± 2.768, respectively. Optimum reducing activity was detected at 72 h of incubation, and MRS broth is a suitable medium. Phylogenetic analysis based on 16S rRNA gene nucleotide sequences confirmed that one of the bacterial isolate RS-5 bacterial isolates with higher mycotoxin degradation is identified as Bacillus subtilis isolated from rumen sample. B05 (FSL-8) bacterial isolate of yogurt belongs to the genus Lactobacillus with 99.66% similarity with Lactobacillus delbrukii. Similarly, three other bacterial isolates, D05, H05 and F04 (FS-17, FSL-2 and FS-20), were found to be the sub-species/strains Pseudomonas gessardii of genus Pseudomonas based on their similarity level of (99.2%, 96% and 96.88%) and positioning in the phylogenetic tree. Promising detoxification results were revealed through GC-MS analysis of RS-5 and FSL-8 activity.     

Fucoidan induces apoptosis of HepG2 cells by down-regulating p-Stat3

Summary： Fucoidan is one of the main bioactive components of polysaccharides. The current study was focused on the anti-tumor effects of fucoidan on human heptoma cell line HepG2 and the possible mechanisms. Fucoidan treatment resulted in cell cycle arrest and apoptosis of HepG2 cells in a dose-dependent manner detected by MTT assay, flow cytometry and fluorescent microscopy. The results of flow cytometric analysis revealed that fucoidan induced G2/M arrest in the cell cycle progression. Hoechst 33258 and Annexin V/PI staining results showed that the apoptotic cell number was increased, which was associated with a dose-dependent up-regulation of Bax and down-regulation of Bcl-2 and p-Stat3. In parallel, the up-regulation of p53 and the increase in reactive oxygen species were also observed, Which may play important roles in the inhibition of HepG2 growth by fucoidan. In the meantime, Cyelin B 1 and CDK1 were down-regulated by fucoidan treatment. Down-regulation of p-Stat3 by fucoidan resulted in apoptosis and an increase in ROS in response to fucoidan exposure. We therefore concluded that fucoidan induces apoptosis through the down-regulation of p-Stat3. These results suggest that fucoidan may be used as a novel anti-cancer agent for hepatocarcinoma.     

Molecularly imprinted polymeric coatings for sensitive and selective gravimetric detection of artemether

Monitoring antimalarial drugs is necessary for clinical assays, human health, and routine quality control practices in pharmaceutical industries. Herein, we present the development of sensor coatings based on molecularly imprinted polymers (MIPs) combined with quartz crystal microbalance (QCM) for sensitive and selective gravimetric detection of an antimalarial drug: artemether. The MIP coatings are synthesized by using artemether as the template in a poly(methacrylic acid-co-ethylene glycol dimethacrylate) matrix. Artemether-MIP and the non-imprinted polymer (NIP) control or reference layers are deposited on 10 MHz dual-electrode QCM by spin coating (187 ± 9 nm layer thickness after optimization). The coatings are characterized by FTIR spectroscopy and atomic force microscopy that reveal marked differences among the MIP and NIP. The MIP-QCM sensor exhibits high sensitivity (0.51 Hz ppm⁻¹) with sub-10 ppm detection and quantification limits. The MIP-QCM sensor also exhibits a 6-fold higher sensitivity compared to the NIP-QCM, and a dynamic working range of 30–100 ppm. The response time of MIP-QCM devices for a single cycle of analyte adsorption, signal saturation, and MIP regeneration is less than 2.5 min. The sensor also demonstrates selectivity factors of artemether-MIP of 2.2 and 4.1 compared to artemisinin and lumefantrine, respectively. Reversibility tests reveal less than 5% variation in sensor responses over three cycles of measurements at each tested concentration. The MIP-QCM showed lower detection limits than conventional HPLC-UV, and faster response time compared to HPLC-UV and liquid chromatography-mass spectrometry (LC-MS).

Chemical structures of the antimalarial drugs: artemisinin, artemether (a methyl ether derivative of artemisinin), and lumefantrine.     

A comprehensive Y-STR portrait of Yousafzai’s population

In the current study, 17 Y-Chromosomal short tandem repeats (Y-STRs) included in theAmpFlSTR Y-Filer amplification kit (Applied Biosystems, Foster City, USA) were investigated in 146 unrelated Yousafzai males residing in the Khyber Pakhtunkhwa Province of Pakistan. A total of 94 (89.52%) unique haplotypes were observed. Discrimination capacity was 71.92%. Haplotype diversity ranged from 0.354 (DYS456) to 0.663 (DYS458). Both Rst pairwise analysis and multidimensional scaling plot showed that the genetic structure of the Yousafzais is significantly different from neighbouring populations.