The energy metabolism of the freshwater leech Whitmania pigra in response to fasting.

Non-blood-feeding leeches, Whitmania pigra, have evolved unique digestive structures and physiological mechanisms to cope with fasting. However, the metabolic changes and molecular mechanisms induced by fasting remain unclear. Therefore, this study recorded the weights of leeches during the fasting process. The weight changes were divided into two stages: a rapid decline period (1-9 weeks) and a fluctuating decline period (9-24 weeks). Leeches fasted for 4 (H4), 11 (H11), and 24 (H24) weeks were selected for transcriptome sequencing. Compared to the control group (H0), 436, 1157, and 337 differentially expressed genes (DEGs) were identified, which were mainly related to glycolysis/gluconeogenesis, amino acid metabolism, and the lipid metabolism pathway. The 6-phosphofructokinase (Pfk), pyruvate kinase (PK), and phosphoenolpyruvate carboxykinase (Pck) transcription levels revealed glycolysis/gluconeogenesis activation during the early stage of fasting and peaked at 11 weeks. Decreased expression of the rate-limiting enzyme acetyl-CoA carboxylase (ACC) in fatty acid synthesis during fasting may impede fatty acid synthesis. These results indicated that the nutrient storage and energy-supplying pathways in W. pigra were modified to improve fasting resistance. The findings of this study provided guidance for exploring the mechanism underlying fasting metabolism and laid a foundation for artificial breeding to improve the resistance of leeches.

Complete mitochondrial DNA sequence of Alboglossiphonia lata Oka, 1910 (Rhynchobdellida: Glossiphoniidae) and its phylogenetic analysis.

The complete mitochondrial genome of Alboglossiphonia lata (basionym: Glossiphonia lata), sourced from a biodiversity hotspot of China, has been determined and reported in this study. It was 15,236 bp in length and consisted of 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and three control regions. The mitogenome was deposited GenBank under the accession number PP165800. A. lata and other species within the Glossiphoniidae family were clustered together with high bootstrap values. The mitochondrial genome of A. lata provides valuable molecular data for further phylogenetic research on the Glossiphoniidae family.

Differential mRNA and lncRNA Expression Profiles Associated with Early Pregnancy Loss in ART Patients.

Early pregnancy loss (EPL) is the most common complication in assisted reproductive technology (ART). However, the precise causes for nearly 50% patients remain unexplained. In the current study, we aimed to discover the differentially expressed profiling of mRNAs and lncRNAs by RNA sequencing (RNA-seq). Human chorionic villi tissues were collected from patients with EPL and natural control (NC) group. RNA sequencing (RNA-seq) of these specimens was performed for transcriptome analysis. As a result, we identified a total of 141 mRNAs and 137 lncRNAs that were significantly differentially expressed between villi tissues from EPL and NC. Functional enrichment analyses indicated enrichment of differentially expressed genes involved in pathways were associated with growth hormone receptor binding, PI3K-Akt signaling pathway, Jak-STAT signaling pathway, transcriptional misregulation in cancer, metabolic pathways and Rap1 signaling pathway. Additionally, the co-expression networks (lncRNA-miRNA-mRNA) was constructed based on the correlation analysis between the differentially expressed RNAs.7 mRNAs and 6 lncRNAs were successfully technically validated with RT-PCR. In conclusion, our results suggest a direction for the further study of EPL-related mRNAs and lncRNAs and may ultimately assist in understanding the pathogenesis of EPL.

Candida causes recurrent vulvovaginal candidiasis by forming morphologically disparate biofilms on the human vaginal epithelium.

Background: Recurrent vulvovaginal candidiasis (RVVC) is a recalcitrant medical condition that affects many women of reproductive age. The importance of biofilm formation by Candida in RVVC has been recently questioned. This study aimed to elucidate the fundamental growth modes of Candida in the vagina of patients with RVVC or sporadic vulvovaginal candidiasis (VVC) and to assess their roles in the persistence of RVVC. Methods: Vaginal tissues were sampled from twelve patients clinically and microbiologically diagnosed as RVVC or VVC at a post-antifungal-treatment and asymptomatic period. High-resolution scanning electron microscopy, fluorescence in situ hybridization in combination with Candida-specific 18S rRNA probes and viable fungal burden were used to qualitatively and quantitatively evaluate Candida growth in the human vagina. The presence of Candida biofilm extracellular polymeric substances was examined using confocal laser scanning microscopy and biopsy sections pre-stained with Concanavalin A. Histopathological analysis was carried out on infected vaginal tissues stained with hematoxylin and eosin. Lastly, the susceptibility of epithelium-associated Candida biofilms to fluconazole at the peak serum concentration was evaluated. Results: Candida species grew on the vaginal epithelium of RVVC patients as morphologically disparate biofilms including monolayers, microcolonies, and macro-colonies, in addition to sporadic adherent cells. Candida biofilm growth on the vaginal epithelium was associated with mild lymphocytic infiltration of the vaginal mucosa. These epithelium-based Candida biofilms presented an important characteristic contributing to the persistence of RVVC that is the high tolerance to fluconazole. Conclusions: In summary, our study provides direct evidence to support the presence of Candida biofilms in RVVC and an important role of biofilm formation in disease persistence.

Division of developmental phases of freshwater leech Whitmania pigra and key genes related to neurogenesis revealed by whole genome and transcriptome analysis.

The freshwater leech Whitmania pigra (W. pigra) Whitman (Annelida phylum) is a model organism for neurodevelopmental studies. However, molecular biology research on its embryonic development is still scarce. Here, we described a series of developmental stages of the W. pigra embryos and defined five broad stages of embryogenesis: cleavage stages, blastocyst stage, gastrula stage, organogenesis and refinement, juvenile. We obtained a total of 239.64 Gb transcriptome data of eight representative developmental phases of embryos (from blastocyst stage to maturity), which was then assembled into 21,482 unigenes according to our reference genome sequenced by single-molecule real-time (SMRT) long-read sequencing. We found 3114 genes differentially expressed during the eight phases with phase-specific expression pattern. Using a comprehensive transcriptome dataset, we demonstrated that 57, 49 and 77 DEGs were respectively related to morphogenesis, signal pathways and neurogenesis. 49 DEGs related to signal pathways included 30 wnt genes, 14 notch genes, and 5 hedgehog genes. In particular, we found a cluster consisting of 7 genes related to signal pathways as well as synapses, which were essential for regulating embryonic development. Eight genes cooperatively participated in regulating neurogenesis. Our results reveal the whole picture of W. pigra development mechanism from the perspective of transcriptome and provide new clues for organogenesis and neurodevelopmental studies of Annelida species.

Independent value of serum ß-human chorionic gonadotropin in predicting early pregnancy loss risks in IVF/ICSI cycles.

Background: Early pregnancy loss (EPL) is the most prevalent complication, particularly in couples undergoing assisted reproductive technology treatment. The present study aimed to determine whether the serum ß-human chorionic gonadotropin (ß-hCG) level after 14 days of embryo transfer, either alone or in conjunction with other parameters in IVF/ICSI cycles, could be used to predict subsequent EPL. Methods: This was a retrospective cohort study of all couples who received clinical pregnancy and underwent fresh IVF/ICSI cycles at a single large reproductive medical center between January 2013 and June 2020. The research involved a total of 6600 cycles. For risk variables, we conducted the least absolute shrinkage and selection operator (LASSO) analysis, and for risk scoring, we used logistic regression coefficients. To analyze relevant risk factors for EPL, univariate and multivariate logistic regression analyses were employed. Areas under the curve (AUC) were determined and compared between ß-hCG and other factors using receiver operating characteristic (ROC) curves. Results: ß-hCG level was considerably lower in women who had EPL than in those who were ongoing pregnancy (564.03 ± 838.16 vs 1139.04 ± 1048.72 IU/L, p< 0.001). Univariable and multivariable logistic regression revealed that ß-hCG levels were significantly correlated with the probability of EPL, independent of other risk factors. More importantly, the ß-hCG level could independently predict the occurrence of EPL and was comparable to the model that combined other risk factors. The optimal serum ß-hCG cut-off value for predicting EPL was 542.45 IU/L. Conclusions: Our results suggest that the serum ß-hCG level has a strong independent predictive value for EPL occurrence in fresh IVF/ICSI cycles.

Risk factors related to early pregnancy loss in fresh IVF/ICSI: An analysis of 954 embryo transfer cycles.

Early pregnancy loss (EPL) is a common complication of assisted reproductive technology treatment; however, the exact factors involved in EPL are not fully understood. This study aimed to evaluate the risk factors for EPL in fresh in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) cycles. This retrospective cohort study was conducted on all couples who underwent clinical pregnancy in fresh IVF/ICSI cycles from January to December 2019 at a single large reproductive medical center. In total, 954 cycles were included in this study. Univariate and multivariate logistic regression analyses were performed to evaluate relevant risk factors for EPL. Curve fitting and threshold analyses were used to explore the association between risk factors and EPL. Compared with women with a normal total antral follicle count (AFC) (≥10, <15), those with a low AFC (<10) had a higher risk of EPL (odds ratio 2.97, 95% confidence interval: 1.38-6.38, P < .05). Patients with an estradiol/progesterone ratio (E2/P) ≥ 1.1 had significantly lower odds of EPL than women with E2/P < 1.1 (odds ratio 0.51, 95% confidence interval 0.28-0.91, P < .05). E2/P and serum human chorionic gonadotropin (hCG) levels negatively correlated with EPL. By using a 2-piecewise regression model, the inflection point of serum hCG level was 599.9 IU/L. Our results showed that lower AFC, E2/P, and serum hCG levels were associated with a higher EPL risk in fresh IVF/ICSI cycles.

Temporal Relation Extraction with Joint Semantic and Syntactic Attention.

Determining the temporal relationship between events has always been a challenging natural language understanding task. Previous research mainly relies on neural networks to learn effective features or artificial language features to extract temporal relationships, which usually fails when the context between two events is complex or extensive. In this paper, we propose our JSSA (Joint Semantic and Syntactic Attention) model, a method that combines both coarse-grained information from semantic level and fine-grained information from syntactic level. We utilize neighbor triples of events on syntactic dependency trees and events triple to construct syntactic attention served as clue information and prior guidance for analyzing the context information. The experiment results on TB-Dense and MATRES datasets have proved the effectiveness of our ideas.

Antibacterial and antibiofilm activities of thyme oil against foodborne multiple antibiotics-resistant Enterococcus faecalis.

The inhibitory and bactericidal activities of thyme oil against the foodborne multiple antibiotics-resistant Enterococcus faecalis biofilm were evaluated in this study. Gas chromatography-mass spectrometry revealed that more than 70% of the composition of thyme oil is thymol. Crystal violet staining assay showed that 128 and 256 µg/mL thyme oil significantly inhibited the biofilm formation of E. faecalis. The cell adherence of E. faecalis, as shown by its swimming and swarming motilities, was reduced by thyme oil. The exopolysaccharide (EPS) quantification assay showed that thyme oil inhibited the EPS synthesis in E. faecalis biofilms. The 3D-view observations through confocal laser scanning and scanning electron microscopy suggested that cell adherence and biofilm thickness were decreased in thyme oil-treated biofilms. Quantitative real-time analyses showed that the transcription of ebp and epa gene clusters, which were related to cell mobility and EPS production, was inhibited by thyme oil. Thus, thyme oil effectively inhibited the biofilm formation of E. faecalis by affecting cell adherence and EPS synthesis. Furthermore, 2,048 and 4,096 µg/mL thyme oil can effectively inactivate E. faecalis population in the mature E. faecalis biofilms by 5.75 and 7.20 log CFU/mL, respectively, after 30 min of treatment. Thus, thyme oil at different concentrations can be used as an effective antibiofilm or germicidal agent to control E. faecalis biofilms.

Antimicrobial Activity of Phenyllactic Acid Against Enterococcus faecalis and Its Effect on Cell Membrane.

3-Phenyllactic acid (PLA) was reported to have an effective antimicrobial activity. This study evaluated the antimicrobial activity of PLA against foodborne Enterococcus faecalis and its effect on cell membrane. The minimum concentration of PLA to inactivate E. faecalis in brain heart infusion broth was 5 mg/mL. PLA solutions of 5 and 10 mg/mL can inactivate E. faecalis population ≥6 log CFU/mL within 60 and 30 min, respectively. The cell membranes of most E. faecalis cells were damaged after PLA treatment according to the images of scanning electron microscopy and transmission electron microscopy. The differences in the regions of cell membrane protein, fatty acid, and polysaccharide were revealed by Fourier transform infrared spectroscopy, which further indicated cell membrane damages. The cell membrane permeability was increased when the concentration of PLA treatment was increased in the membrane permeability assays. Finally, almost all bacterial cells were damaged after treatment with 10 mg/mL PLA for 30 min, further confirmed by flow cytometry analysis. This study concluded that PLA is effective in inactivating E. faecalis cells through the leakage of intracellular components caused by cell membrane damage.

Altered Circular RNA Expression in Patients with Repeated Implantation Failure.

BACKGROUND/AIMS: CircRNAs play an important role in regulating gene expression and the specific role of circRNAs in the pathogenesis of repeated implantation failure remains unclear. The aim of this study is to assess the differentially expressed circRNAs in patients with repeated implantation failure. METHODS: We screened circRNA expression profiles in endometrial biopsies taken from six women with repeated implantation failure and control group using circRNA microarray. Bioinformatic analyses were applied to study these differentially expressed circRNAs. Furthermore, quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to confirm these results. RESULTS: The data from circRNA microarrays clearly revealed that 856 unique circRNAs were significantly altered (p<0.05). The up-regulated expression of hsa_circRNA_070616, hsa_circRNA_103716, hsa_circRNA_104001, hsa_circRNA_104854 and the down-regulated expression of hsa_circRNA_004183, hsa_circRNA_044353, hsa_circRNA_404686 were further validated by qRT-PCR. CONCLUSION: this study demonstrates that a number of circRNAs were differentially expressed in patients with repeated implantation failure compared with normal controls and may offer novel molecular candidates for diagnosis and clinical treatment of embryo implantation failures.

A high surface area flower-like Ni-Fe layered double hydroxide for electrocatalytic water oxidation reaction.

Layered double hydroxide has been used in a variety of areas, including but not limited to catalysis, energy storage, drug or gene delivery, water treatment, etc. Herein, we report a new simple hydrothermal method to prepare a high surface area flower-like Ni-Fe layered double hydroxide (LDH) assembled by nanosheets by using nickel alkoxide and FeSO4 as the only starting materials. It is free of alkaline solution and other additives for directing or supporting in the synthesis procedure. The formation mechanism of this flower-like LDH formed by ultrathin nanosheets is also discussed. Moreover, the as-obtained LDH material shows increased electrocatalytic activity and stability toward WOR in alkaline media compared with the materials prepared without a Ni alkoxide precursor or Fe precursor, namely α-Fe2O3 and Ni(OH)2, respectively. In addition, the electrocatalytic activity is demonstrated to be related to the molar ratio of Fe and Ni in the final Ni-Fe material, and the best activity is achieved when the ratio reaches 0.52 : 1. The phase compositions of the resulting Ni-Fe(x) are discussed. Furthermore, the Ni-Fe LDH material reported herein might be employed as a promising noble-metal-free water oxidation catalyst to replace the IrOx material-the state-of-the-art water oxidation catalyst.

Precursor-mediated synthesis and sensing properties of wurtzite ZnO microspheres composed of radially aligned porous nanorods.

Wurtzite ZnO microspheres composed of radially aligned porous nanorods are prepared via a simple thermal treatment of a "pre-synthesized" zinc monoglycerolate precursor. The as-prepared hierarchical nanomaterial can serve as a highly sensitive sensing material for ethanol detection.