The mediation of the placenta on the association between maternal ambient temperature exposure and birth weight.

Studies have indicated that exposure to low and high temperatures during pregnancy negatively affects fetal development. The placenta plays vital functions in fetal development and could also be impacted by suboptimal temperatures. However, whether the placenta mediates the association between suboptimal temperature and birth weight is unknown. Our study aims to evaluate the association between ambient temperature and birth weight as well as the mediation effect of the placenta. A prospective birth cohort study was conducted during 2017-2020 in Guangzhou, China (n = 3349 participants). We defined extreme temperature exposure during the whole pregnancy by using different thresholds, including low temperatures (< 25th, < 15th, < 10th, < 5th percentiles), and high temperatures (> 75th, > 85th, > 90th, > 95th percentiles). Three different approaches (generalized linear model, inverse probability weighting, and doubly robust model) were applied to estimate the effects of low/high temperatures on birth weight and placental indicators, including placental weight, placental volume, and placental-to-birth weight ratio (PFR), respectively. We observed that both low and high ambient temperatures during the whole pregnancy were associated with lower birth weight and negative changes in placental indicators. The estimated lower mean birth weight ranged from -158 g (95 % CI: -192 g, -123 g) to -363 g (95 % CI: -424 g, -301 g) for low temperatures and from -97 g (95 % CI: -135 g, -59 g) to -664 g (95 % CI: -742 g, -585 g) for high temperatures. In mediation analyses, placental weight mediated 28.79 % to 40.47 % and 48.22 % to 54.38 % of the association of low and high temperatures with birth weight, respectively. The findings suggest that placental weight may mediate the association between ambient temperature exposure and birth weight.

Identifying the critical windows and joint effects of temperature and PM2.5 exposure on small for gestational age.

The potential critical windows for extreme ambient temperature, air pollution exposure and small for gestational age (SGA) are still unclear, and no study has explored their joint effects on SGA. In a national multi-center prospective cohort, we included 179,761 pairs of mother-infant from 16 counties of 8 provinces in China during 2014-2018. Daily averaged temperature and PM2.5 concentration were matched to the maternal residential address to estimate personal exposure. Extreme temperature exposures were categorized by a series of percentile in each meteorological and geographic division for the entire pregnancy, each trimester and gestational week (GA-week). Generalized linear mixed models (GLMMs) and distributed lag nonlinear models (DLNMs) were used to estimate the whole pregnancy-, trimester-specific, and weekly-specific associations of extreme temperature and PM2.5 exposures with SGA. Combined effects were evaluated with the relative excess risk due to interaction (RERI) and proportion attributable to interaction (AP). We observed that by referring to temperature at the 41st - 50th percentile, heat (>90th percentile) exposure during 13th - 29th GA-weeks was associated with SGA; odds ratio (OR) and 95 % confidence intervals (CI) was 1.16 (1.06, 1.28). For cold (<=10th percentile), inverse associations were observed during the 1st - 8th GA-weeks. PM2.5 exposure during the 2nd - 5th and 19th - 27th GA-weeks was associated with SGA, with the strongest association in the 2nd GA-week (OR = 1.0017, 95 %CI: 1.0001, 1.0034, for a 10 µg/m3 increase). No interactive effects between ambient temperature and PM2.5 on SGA were observed. Our findings suggest the weekly susceptibility windows for heat and PM2.5 exposure were primarily the gestational weeks within the 2nd trimester, therefore, corresponding protective measures should be conveyed to pregnant women during routine prenatal visits to reduce exposures.

Thermal perception and lung function: a panel study in young adults with exercise under high outdoor temperature.

It has been observed that high temperature exposure is associated with a reduction in lung function and some possible biological mechanisms have been suggested. However, it is unclear if thermal perception plays a role in the association. From September 3rd to 15th, 2018, in Guangzhou, China, we repeatedly measured daily thermal perception and lung function among 126 participants with outdoor military training. We performed a linear mixed model and stratified analyses by the origin of students, gender, and the training period to evaluate the effects of thermal perception on lung function. A total of 399 measurements were collected. Per vote increase in thermal sensation vote towards the "hot" direction was associated with a - 0.04 L (95% CI: - 0.08 to - 0.01) decrease in forced vital capacity (FVC), and - 0.04 L (95% CI: - 0.08 to - 0.01) decrease in forced expiratory volume in 1 s (FEV1). Per grade increase towards the "very uncomfortable" direction for thermal comfort vote was associated with an increased percentage of forced expiratory volume in 1 s (FEV1%) by 1.52% (95% CI: 0.18 to 2.86). For thermal preference, with preferred cooler vote increased by one level, FVC and FEV1 decreased by - 0.05 L/s (95% CI: - 0.08 to - 0.02) and - 0.05L/s (95% CI: - 0.08 to - 0.02), respectively. The effects of thermal perception on lung function were stronger among non-local and in the first week of training. Our study suggests that in the same high-temperature environment, thermal perception is associated with lung function, even in healthy adults.

High concordance in preimplantation genetic testing for aneuploidy between automatic identification via Ion S5 and manual identification via Miseq.

The Ion S5 (Thermo Fisher Scientific) and Miseq (Illumina) NGS systems are both widely used in the clinical laboratories conducting PGT-A. Each system employs discrepant library preparation steps, sequencing principles, and data processing algorithms. The automatic interpretation via Ion Reporter software (Thermo Fisher Scientific) and the manual interpretation via BlueFuse Multi software (Illumina) for chromosomal copy number variation (CNV) represent very different reporting approaches. Thus, it is intriguing to compare their ability of ploidy detection as PGT-A/NGS system. In the present study, four aneuploid cell lines were individually mixed with a diploid cell line at different aneuploid ratios of 0% (0:5), 10% (1:9), 20% (1:4), 40% (2:3), 50% (3:3), 60% (3:2), 80% (4:1) and 100% (5:0) to assess the sensitivity and specificity for whole chromosomal and segmental aneuploidy detection. The clinical biopsies of 107 blastocysts from 46 IVF/PGT-A cycles recruited between December 2019 and February 2020 were used to calculate the concordance. Initially, the pre-amplified products were divided into two aliquots for different library preparation procedures of each system. Applying the same calling criteria, automatic identification was achieved through the Ion Reporter, while well-trained technicians manually identified each sample through the BlueFuse Multi. The results displayed that both systems reliably distinguished chromosomal CNV of the mixtures with at least 10% aneuploidy from karyotypically normal samples ([Ion S5] whole-chromosomal duplication: 2.14 vs. 2.05, p value = 0.009, segmental deletion: 1.88 vs. 2.05, p value = 0.003; [Miseq] whole-chromosomal duplication: 2.12 vs. 2.03, p value = 0.047, segmental deletion: 1.82 vs. 2.03, p value = 0.002). The sensitivity and specificity were comparable between the Ion S5 and Miseq ([sensitivity] 93% vs. 90%, p = 0.78; [specificity] 100% vs. 100%, p value = 1.0). In the 107 clinical biopsies, three displayed chaotic patterns (2.8%), which could not be interpreted for the ploidy. The ploidy concordance was 99.04% (103/104) per embryo and 99.47% (2265/2277) per chromosome pair. Since their ability of detection were proven to be similar, the automatic identification in Ion S5 system presents comparatively faster and more standardized performance.

NR1D1 suppressed the growth of ovarian cancer by abrogating the JAK/STAT3 signaling pathway.

BACKGROUND: Nuclear receptor subfamily 1 group D member 1 (NR1D1), a nuclear receptor associated with a variety of physiological processes, has a low level in ovarian cancer tissues compared with adjacent normal tissues. However, its role in ovarian cancer remains unclear. METHODS: The level of NR1D1 in ovarian cancer cells was determined by quantitative real-time PCR. Its role in ovarian cancer was explored through gain-of-function and lose-of-function. Cell growth was evaluated by CCK8 assay, immunofluorescence and flow cytometry. Western blot was conducted to assess the activation of JAK/STAT3 signaling pathway. A xenograft model of ovarian cancer was established to explore the role of NR1D1 in vivo. RESULTS: Up-regulation of NR1D1 repressed the ovarian cancer cell proliferation and induced cell cycle arrest and apoptosis, while silencing NR1D1 promoted their proliferation and G1/S transition. In addition, the JAK/STAT3 signaling pathway, an intracellular signal transduction closely associated with cancer progression, was inhibited by NR1D1. Consistently, xenografts with NR1D1 over-expression grew more slowly in vivo than the controls. Furthermore, NR1D1 up-regulated the expression of suppressor of cytokine signaling 3 (SOCS3), an inhibitor of the JAK/STAT3 signaling pathway. Whereas, SOCS3 silencing abolished the function of NR1D1 over-expression on ovarian cancer growth and JAK/STAT3 signaling pathway. CONCLUSIONS: NR1D1 up-regulated the expression of SOCS3, resulting in suppression of the JAK/STAT3 signaling pathway, thus retarding the growth of ovarian cancer cells. This study highlights a profound role of NR1D1 in the treatment of ovarian cancer.

Hyperoxia sensitizes hypoxic HeLa cells to ionizing radiation by downregulating HIF­1α and VEGF expression.

The current study investigated whether hyperoxia may reverse hypoxia­induced radioresistance (RR) in cervical cancer. Human HeLa cells exposed to hypoxic, normoxic or hyperoxic conditions were irradiated using X­rays. Cell proliferation and apoptosis were analyzed using MTT assays and flow cytometry. The expression levels of hypoxia­inducible factor­1α (HIF­1α), VEGF165, VEGFRs, Akt and ERK were measured via western blotting and/or ELISA. The results demonstrated that hypoxia stimulated HIF­1α and VEGF expression, and induced RR in HeLa cells. The administration of recombinant VEGF or the forced expression of VEGF promoted RR, whereas inactivating HIF­1α or blocking the VEGF­VEGFR interaction abrogated hypoxia­induced RR. Notably, hyperoxia decreased the level of hypoxia­stimulated HIF­1α and VEGF, and enhanced radiosensitivity in hypoxic HeLa cells. The results demonstrated that hyperoxia suppressed the hypoxia­activated Akt and ERK signaling pathways in HeLa cells. Therefore, a high O2 concentration may be considered as a radiotherapeutic sensitizer for hypoxic HeLa cells.

Concordance between different trophectoderm biopsy sites and the inner cell mass of chromosomal composition measured with a next-generation sequencing platform.

STUDY QUESTION: In PGS, does chromosomal constitution differ among trophectoderm (TE) biopsy sites and between them and the inner cell mass (ICM)? SUMMARY ANSWER: The ploidy concordance between ICM and TE was independent of whether the biopsy site in the TE was near to or far from the ICM. WHAT IS KNOWN ALREADY: TE biopsies are considered less harmful to developing embryos than blastomere biopsies. Removal of multi-cellular samples permits high-resolution next-generation sequencing (Veriseq NGS) to detect aneuploidy present in a minority of cells (mosaicism of diploid and aneuploid cells). However, the prevalence of ploidy discrepancies between different TE biopsy sites and the ICM, as well as confined mosaicism (aneuploidy only in a particular area), has not been established. STUDY DESIGN, SIZE, DURATION: Biopsies were taken from a site opposite to the ICM (TE1), near the ICM (TE2) and within the ICM of the same embryo in 33 donated blastocysts obtained from 12 volunteer patients. The samples were analyzed by the Veriseq NGS to assess ploidy concordance. PARTICIPANTS/MATERIALS, SETTING, METHODS: The mean age of the patients was 34.4 years, and samples from all three biopsy sites were achieved in 29 frozen thawed blastocysts. The aneuploid percentage in each sample was interpreted by Veriseq NGS at the finest resolution involving the number of reads after filtering, sample overall noise score, and average quality/alignment scores according to the Veriseq quality control assessment. Ploidy concordance was then assessed between different TE fractions, and between the TE and ICM. MAIN RESULTS AND THE ROLE OF CHANCE: The euploid rates were similar in the TEs and ICM, and no preferential allocation of euploid lineage within a blastocyst was demonstrated. Whether the biopsy site in the TE was near to or far from the ICM, the chromosomal consistency rate was similar [TE1-to-ICM, 86.2% (25/29) versus TE2-to-ICM, 89.7% (26/29); P = 1.0], suggesting that the cells with different chromosomal components may spread randomly throughout the TE. The following two types of inconsistent PGS conclusions between TE and ICM due to confined mosaicism were observed: (i) euploid TE with mosaic ICM (3%) (1/29); and (ii) mosaic TE with euploid ICM (3%) (1/29) or with aneuploid ICM (7%) (2/29). Thus, the overall rate of confined mosaicism was 14% (4/29). LARGE SCALE DATA: N/A. LIMITATION, REASONS FOR CAUTION: The approach used in the present study was affected by biopsy manipulation limitations involving possible cell contamination and the technical challenge of comprehensive chromosomal screening (CCS) procedures. WIDER IMPLICATIONS OF THE FINDINGS: The rate of confined mosaicism in the blastocysts was estimated in this preliminary study, thus, specifying the incidence of biological sampling biases. The results also verified the random distribution of different cell lineages, and the representative value of a single biopsied sample from the TE. STUDY FUNDING AND CONFLICT OF INTEREST(S): No external funding was obtained; all the authors declare no conflicts of interest regarding this study.

Identification of mosaic and segmental aneuploidies by next-generation sequencing in preimplantation genetic screening can improve clinical outcomes compared to array-comparative genomic hybridization.

BACKGROUND: Chromosomal mosaicism is observed as the presence of both euploid and aneuploid cells in a particular blastocyst. Recent studies have reported that the implantation rate of mosaic embryo transfer is remarkably lower than the euploid embryos. The superior capability of next-generation sequencing (NGS) to detect chromosomal mosaicism in preimplantation genetic screening (PGS) remains controversial, and several data displayed similar implantation and pregnancy rates using NGS or array comparative genomic hybridization (aCGH). RESULTS: In this study, the main inconsistency of aneuploidy detection and clinical performance between the NGS and aCGH were assessed. The phase I consisted of a parallel comparison in 182 blastocysts from 45 selected PGS patients for both the NGS and aCGH platforms. The phase II retrospectively compared the clinical outcomes of 90 patients with NGS-screened euploid embryo transfer to that of 129 patients with aCGH-screened euploid embryo transfer. The parallel comparison showed that the inconsistency of embryo euploidy was 11.8% (p = 0.01). Chromosomal mosaicism (10.7% with NGS vs. 3.9% with aCGH) and segmental aneuploidy (10.7% with NGS vs. 6.7% with aCGH) contributed to the discrepancy mainly. The chromosomally mosaic embryos (20%-50% of aneuploidy) and several embryos with segmental aneuploidy (≥10 Mbp) were hard to distinguish using the aCGH platform, but could be clearly identified using the NGS platform. After the first euploid embryo cryotransfer, the ß-HCG(+) rate and implantation rate significantly increased in the PGS/NGS patients (HCG[+] rate: 73.3% in PGS/NGS vs. 60.5% in PGS/aCGH, p = 0.048; implantation rate: 53.2% in PGS/NGS vs. 45.0% in PGS/aCGH, p = 0.043). The clinical and ongoing pregnancy rates appeared higher in the NGS group, but did not reached statistical significance. CONCLUSIONS: The results demonstrated that the NGS platform can identify embryos with chromosomal mosaicism and segmental aneuploidy more precisely than the aCGH platform, and the following clinical performance of NGS was more favorable.

Anti-herpes simplex virus activity of polysaccharides from Eucheuma gelatinae.

Acyclovir is a commonly-used drug for treating herpes simplex virus (HSV) infections, but with its wide clinical application, more and more resistant strains have been found. Therefore, seeking a drug that can act against acyclovir-resistant virus has become an important goal of drug screening and development. In this study, plaque reduction assay, real-time PCR, Western blot, and immunofluorescence technique were used to investigate the antiviral effect of Eucheuma gelatinae polysaccharide (EGP) on HSV and to preliminarily clarify the in vitro anti-HSV mechanism of EGP. EGP was found to significantly inhibit HSV infection in vitro and displayed a good inhibitory effect on acyclovir-resistant strains. More detailed experiments have shown that EGP prevented early HSV-1 infection through directly inactivating HSV-1 particles and impairing virus attachment, but without effect on viral penetration. EGP also inhibited the RNA synthesis of HSV-1 early gene and late gene as well as viral DNA replication; no effect on immediate-early gene synthesis was observed. Besides, through immunofluorescence and western blot, we found that EGP significantly affected the protein synthesis of HSV-1. Taken together, these results demonstrate that EGP exerts its anti-HSV activity mainly through impeding early HSV-1 infection and inhibiting viral RNA and DNA syntheses. The weak cytotoxicity, strong viral inactivation as well as attachment inhibition activity enable EGP to be a virucide candidate for HSV therapy, especially for drug-resistant strains.

[Progress in polyphosphate and related metabolizing enzymes].

Inorganic polyphosphate (Poly P) is a polymer consisting of ten to hundreds of phosphate residues linked by "high-energy" phosphoanhydride bonds, which is abundantly found in all organisms and nature. Here the basic facts of PolyP are summarized: genes regulated by polyP, role in DNA uptake, motility of microorganism, function in stress response, the virulence of pathogens, as well as the proliferation of mammary cancer cells, blood coagulation, cell calcification and the modulation of mitochondrial activity. Enzymes with activities requiring polyP, such as endopolyphosphatase, glucokinase, NAD kinase, AMP phosphotransferase are outlined too. The structure and activity of enzymes regulating polyP level such as polyphosphate kinase and exopolyphosphatase are noted. A thorough analysis of the mycobacterium tuberculosis PPX protein homologs and their biochemical activity is presented.