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Chemosphere Jul 2024The resuspension of phosphorus (P) in sediments has the most significant contribution to the overlying water. The PP release characterization during resuspension was...
The resuspension of phosphorus (P) in sediments has the most significant contribution to the overlying water. The PP release characterization during resuspension was investigated. The results indicated that the P in suspensions had more release risk compared to the sediments. The particulate P (PP) concentration (0.54 mg L) under high-intensity rotational speed (250 rad min) was about five times higher than others (0.11 mg L). The sorption parameters of zero equilibrium P concentration (EPC) and soluble reactive P (SRP) were significantly correlated with each other (p < 0.01, r = 0.73). Suspended solids expressed stronger P source than sediments. The values of EPC was highly significantly correlated with the sorption coefficient (K) and native adsorbed P (NAP) (p < 0.01). The mean values of NAP were 0.0612 mg g and 0.0604 mg g in the Prophase and Metaphase, respectively, and 0.0586 mg g at Anaphase. The values of P sorption index (PSI) ranged from 0.4359 to 0.6862 L g, with mean values of 0.5350 L g (Prophase), 0.6061 L g (Metaphase), and 0.4967 L g (Anaphase). The degree of P saturation (DPS) decreased in the order of Anaphase (2.73%) > Prophase (2.53%) > Metaphase (2.12%). The release risk index of P (ERI) decreased in the order of Anaphase (5.47%) > Prophase (4.72%) > Metaphase (3.59%), with a range of 2.12%-8.56%. To fast and slow scale, the results of NaOH-P (V<0, V>0) contribution indicated that the persistent disturbance promoted the release of adsorbed dissolved PP from NaOH-P in suspended sediment to the overlying water. The contribution of HCl-P (V > 0) was positive in the Anaphase of the slow scale, and HCl-P was a PP source in the frequently disturbing conditions.
Topics: Phosphorus; Geologic Sediments; Water Pollutants, Chemical; Environmental Monitoring; Adsorption
PubMed: 38777196
DOI: 10.1016/j.chemosphere.2024.142386 -
Computers in Biology and Medicine Jul 2024Automated karyotyping is of great importance for cytogenetic research, as it speeds up the process for cytogeneticists through incorporating AI-driven automated...
KaryoXpert: An accurate chromosome segmentation and classification framework for karyotyping analysis without training with manually labeled metaphase-image mask annotations.
Automated karyotyping is of great importance for cytogenetic research, as it speeds up the process for cytogeneticists through incorporating AI-driven automated segmentation and classification techniques. Existing frameworks confront two primary issues: Firstly the necessity for instance-level data annotation with either detection bounding boxes or semantic masks for training, and secondly, its poor robustness particularly when confronted with domain shifts. In this work, we first propose an accurate segmentation framework, namely KaryoXpert. This framework leverages the strengths of both morphology algorithms and deep learning models, allowing for efficient training that breaks the limit for the acquirement of manually labeled ground-truth mask annotations. Additionally, we present an accurate classification model based on metric learning, designed to overcome the challenges posed by inter-class similarity and batch effects. Our framework exhibits state-of-the-art performance with exceptional robustness in both chromosome segmentation and classification. The proposed KaryoXpert framework showcases its capacity for instance-level chromosome segmentation even in the absence of annotated data, offering novel insights into the research for automated chromosome segmentation. The proposed method has been successfully deployed to support clinical karyotype diagnosis.
Topics: Humans; Karyotyping; Metaphase; Algorithms; Chromosomes, Human; Image Processing, Computer-Assisted; Deep Learning
PubMed: 38776728
DOI: 10.1016/j.compbiomed.2024.108601 -
Molecular Human Reproduction May 2024The role of cumulus cells (CCs) in the acquisition of oocyte developmental competence is not yet fully understood. In a previous study, we matured cumulus-denuded...
The role of cumulus cells (CCs) in the acquisition of oocyte developmental competence is not yet fully understood. In a previous study, we matured cumulus-denuded fully-grown mouse oocytes to metaphase II (MII) on a feeder layer of CCs (FL-CCs) isolated from developmentally competent (FL-SN-CCs) or incompetent (FL-NSN-CCs) SN (surrounded nucleolus) or NSN (not surrounding nucleolus) oocytes, respectively. We observed that oocytes cultured on the former could develop into blastocysts, while those matured on the latter arrested at the 2-cell stage. To investigate the CC factors contributing to oocyte developmental competence, here we focused on the CCs' release into the medium of extracellular vesicles (EVs) and on their miRNA content. We found that, during the 15-h transition to MII, both FL-SN-CCs and FL-NSN-CCs release EVs that can be detected, by confocal microscopy, inside the zona pellucida (ZP) or the ooplasm. The majority of EVs are <200 nm in size, which is compatible with their ability to cross the ZP. Next-generation sequencing of the miRNome of FL-SN-CC versus FL-NSN-CC EVs highlighted 74 differentially expressed miRNAs, with 43 up- and 31 down-regulated. Although most of these miRNAs do not have known roles in the ovary, in silico functional analysis showed that seven of these miRNAs regulate 71 target genes with specific roles in meiosis resumption (N = 24), follicle growth (N = 23), fertilization (N = 1), and the acquisition of oocyte developmental competence (N = 23). Overall, our results indicate CC EVs as emerging candidates of the CC-to-oocyte communication axis and uncover a group of miRNAs as potential regulatory factors.
Topics: Animals; Cumulus Cells; Extracellular Vesicles; Oocytes; MicroRNAs; Mice; Female; In Vitro Oocyte Maturation Techniques; Oogenesis; Zona Pellucida
PubMed: 38745364
DOI: 10.1093/molehr/gaae019 -
Cell Journal May 2024This study aimed to investigate the relationship between follicular fluid Bisphenol A (BPA) concentrations with alterations in the expressions of and genes and the...
OBJECTIVE
This study aimed to investigate the relationship between follicular fluid Bisphenol A (BPA) concentrations with alterations in the expressions of and genes and the number of retrieved mature oocytes (MII oocyte) in the cumulus cells of infertile poor ovarian response stimulates women.
MATERIALS AND METHODS
In this prospective cohort study, 80 infertile unexpected poor ovarian response (POR) subjects were selected on the basis of subgroup 1a of the POSEIDON classification. They were divided into two groups: group 1 consisted of 40 women, each with a higher number of metaphase II (MII) oocytes (G1, 3-4 oocytes retrieved), while group 2 comprised of 40 women, each with a lower number of MII oocytes (G2, ≤2 oocytes retrieved). The expressions of the studied genes were evaluated by quantitative-real time polymerase chain reaction (PCR). The concentration of BPA in follicular fluid was measured with HPLC.
RESULTS
The expression levels of , and genes were significantly lower in G2 than G1 (P<0.05). Meanwhile, expression levels were higher in unexpected POR patients in G2 compared to G1 (P<0.05). There was a significant direct correlation between the levels of and gene expressions and there was also a significant inverse correlation (P<0.05) between the levels of CASPASE 3/7, with the number of MII oocytes and embryo development between the two groups. The concentration of BPA in the follicular fluids of G2 was higher compared to G1 (P<0.05).
CONCLUSION
A higher concentration of BPA was associated with a lower number of mature oocytes and oocyte quality in these patients. Also, alterations of and transcript levels in unexpected POR women were associated with BPA concentration.
PubMed: 38736411
DOI: 10.22074/cellj.2024.2020628.1488 -
Reproductive Biomedicine Online Mar 2024Can immature oocytes vitrified and warmed using a short protocol survive and resume meiosis?
RESEARCH QUESTION
Can immature oocytes vitrified and warmed using a short protocol survive and resume meiosis?
DESIGN
This study examined modifications of oocyte vitrification and warming protocols that reduce the length of exposure to vitrification and warming solutions. In total, 561 germinal vesicles and 218 metaphase I oocytes that were immature at oocyte retrieval were vitrified at room temperature for 2 min. Warming was performed at 37°C for 2 min. Resumption of meiotic activity was evaluated after 24 and 48 h of culture. Two different commercially available vitrification and warming kits were used for comparison.
RESULTS
Ninety-five percent of germinal vesicles survived, with no difference observed between the kits. The survival of metaphase I oocytes was, on average, 95.4% and did not differ significantly between the kits. Of the 533 germinal vesicles that survived, 491 converted to metaphase I oocytes (92.1%). After culture for 48 h, 54.4% converted to metaphase II oocytes. In addition, of the 208 metaphase I oocytes that survived warming, 84.1% converted to metaphase II oocytes after 24 h of culture. These maturation rates were similar to those of non-vitrified oocytes.
CONCLUSIONS
Vitrification and warming of oocytes at different nuclear maturation stages can be performed with 2 min of exposure to hypertonic solution and 2 min of exposure to hypotonic solution, respectively. This approach reduces exposure of the oocytes to room temperature during dehydration and rehydration. Warming in 0.5M sucrose helps to maintain and support the potential of oocytes to resume nuclear meiotic activity, and conversion from germinal vesicles to metaphase I and metaphase II oocytes.
PubMed: 38733676
DOI: 10.1016/j.rbmo.2024.103976 -
International Journal of Molecular... Apr 2024Numerous post-translational modifications are involved in oocyte maturation and embryo development. Recently, lactylation has emerged as a novel epigenetic modification...
Numerous post-translational modifications are involved in oocyte maturation and embryo development. Recently, lactylation has emerged as a novel epigenetic modification implicated in the regulation of diverse cellular processes. However, it remains unclear whether lactylation occurs during oocyte maturation and embryo development processes. Herein, the lysine lactylation (Kla) modifications were determined during mouse oocyte maturation and early embryo development by immunofluorescence staining. Exogenous lactate was supplemented to explore the consequences of modulating histone lactylation levels on oocyte maturation and embryo development processes by transcriptomics. Results demonstrated that lactylated proteins are widely present in mice with tissue- and cell-specific distribution. During mouse oocyte maturation, immunofluorescence for H3K9la, H3K14la, H4K8la, and H4K12la was most intense at the germinal vesicle (GV) stage and subsequently weakened or disappeared. Further, supplementing the culture medium with 10 mM sodium lactate elevated both the oocyte maturation rate and the histone Kla levels in GV oocytes, and there were substantial increases in Kla levels in metaphase II (MII) oocytes. It altered the transcription of molecules involved in oxidative phosphorylation. Moreover, histone lactylation levels changed dynamically during mouse early embryogenesis. Sodium lactate at 10 mM enhanced early embryo development and significantly increased lactylation, while impacting glycolytic gene transcription. This study reveals the roles of lactylation during oocyte maturation and embryo development, providing new insights to improving oocyte maturation and embryo quality.
Topics: Animals; Histones; Oocytes; Mice; Embryonic Development; Female; Protein Processing, Post-Translational; Oogenesis; Lysine; In Vitro Oocyte Maturation Techniques; Gene Expression Regulation, Developmental
PubMed: 38732042
DOI: 10.3390/ijms25094821 -
Foods (Basel, Switzerland) Apr 2024As the most consumed tea in the world, all kinds of black tea are developed from Wuyi black tea. In this study, quality components, regulatory gene expression, and key...
As the most consumed tea in the world, all kinds of black tea are developed from Wuyi black tea. In this study, quality components, regulatory gene expression, and key enzyme activity during the processing were analyzed to illustrate the taste formation of WBT. Withering mainly affected the content of amino acids, while catechins and tea pigments were most influenced by rolling and the pre-metaphase of fermentation. Notably, regulatory gene expression was significantly down-regulated after withering except for polyphenoloxidase1, polyphenoloxidase2, leucoanthocyanidin dioxygenase, chalcone isomerase, and flavonoid 3', 5'-hydroxylase. Co-expression of flavonoid pathway genes confirmed similar expression patterns of these genes in the same metabolic pathway. Interestingly, rolling and fermentation anaphase had a great effect on polyphenol oxidase, and fermentation pre-metaphase had the greatest effect on cellulase. Since gene regulation mainly occurs before picking, the influence of chemical reaction was greater during processing. It was speculated that polyphenol oxidase and cellulase, which promoted the transformation of quality components, were the key factors in the quality formation of WBT. The above results provide theoretical basis for the processing of WBT and the reference for producing high-quality black tea.
PubMed: 38731743
DOI: 10.3390/foods13091373 -
The Journal of Cell Biology Aug 2024Accurate chromosome segregation requires sister kinetochores to biorient, attaching to opposite spindle poles. To this end, the mammalian kinetochore destabilizes...
Accurate chromosome segregation requires sister kinetochores to biorient, attaching to opposite spindle poles. To this end, the mammalian kinetochore destabilizes incorrect attachments and stabilizes correct ones, but how it discriminates between these is not yet clear. Here, we test the model that kinetochore tension is the stabilizing cue and ask how chromosome size impacts that model. We live image PtK2 cells, with just 14 chromosomes, widely ranging in size, and find that long chromosomes align at the metaphase plate later than short chromosomes. Enriching for errors and imaging error correction live, we show that long chromosomes exhibit a specific delay in correcting attachments. Using chromokinesin overexpression and laser ablation to perturb polar ejection forces, we find that chromosome size and force on arms determine alignment order. Thus, we propose a model where increased force on long chromosomes can falsely stabilize incorrect attachments, delaying their biorientation. As such, long chromosomes may require compensatory mechanisms for correcting errors to avoid chromosomal instability.
Topics: Animals; Cell Line; Chromosome Segregation; Chromosomes, Mammalian; Kinetochores; Mitosis; Spindle Apparatus; Potoroidae
PubMed: 38727808
DOI: 10.1083/jcb.202310010 -
Human Reproduction (Oxford, England) May 2024Does intraovarian platelet-rich plasma (PRP) injection increase the number of mature oocytes obtained after controlled ovarian stimulation (COS) in young women with poor...
STUDY QUESTION
Does intraovarian platelet-rich plasma (PRP) injection increase the number of mature oocytes obtained after controlled ovarian stimulation (COS) in young women with poor ovarian response (POR) undergoing IVF?
SUMMARY ANSWER
Intraovarian PRP injection procedure does not improve mature oocyte yield after COS in women less than 38 years old with an established IVF history of POR.
WHAT IS KNOWN ALREADY
POR is frequently encountered among the infertile population and the number of women seeking infertility treatment related to POR is increasing. Effective treatment options for this patient population to conceive with autologous oocytes are lacking. Case series and cohort studies suggest that intraovarian PRP injection may improve follicular recruitment in women with premature ovarian insufficiency (POI) and POR, yet robust randomized studies have not been performed to date to determine the clinical utility of this intervention.
STUDY DESIGN, SIZE, DURATION
This was a multi-center randomized controlled trial (RCT) conducted at university-affiliated reproductive centers in the USA and Turkey, between January 2020 and November 2022.
PARTICIPANTS/MATERIALS, SETTING, METHODS
Patients who met inclusion criteria (<38 years old, two or more prior cycles with <3 oocytes retrieved; and without single gene disorders, prior ovarian surgery, endometriomas, BMI >35 kg/m2, or severe male factor infertility) were randomized to either the PRP or control group. Patients in both groups subsequently underwent COS, oocyte retrieval, ICSI, preimplantation genetic testing for aneuploidy (PGT-A), and single euploid embryo transfer. Number of metaphase II (MII) oocytes obtained was the primary outcome. Secondary outcomes included ovarian reserve tests (antral follicle count [AFC] and anti-Müllerian hormone [AMH]), blastocyst and euploid blastocyst yields, and sustained implantation. The study was powered to detect a difference of one mature oocyte obtained at oocyte retrieval.
MAIN RESULTS AND THE ROLE OF CHANCE
In total, 83 patients met inclusion criteria and were randomized to receive autologous intraovarian PRP injection (n = 41) or to no intervention (n = 42). No significant differences were observed in number of MII oocytes retrieved per cycle (2.8 ± 2.4 vs 3.1 ± 3.3 in PRP vs control, respectively; P = 0.9), blastocysts (1.0 ± 1.3 vs 1.3 ± 2.1, P = 0.8), or euploid blastocysts (0.8 ± 1.1 vs 0.9 ± 1.6; P = 0.5). Similarly, no differences were observed in the likelihood of obtaining at least one euploid blastocyst (45% vs 37%, P = 0.4; relative risk [RR], 95% CI = 0.9, 0.6-1.2) or the rate of sustained implantation (31% vs 29%, P = 0.9; RR 1.0, 0.7-1.3). Posttreatment AFC (7.9 ± 4.5 vs 6.8 ± 4.8, P = 0.3) and AMH (0.99 ± 0.98 vs 0.7 ± 0.6, P = 0.2) were also not different between the groups.
LIMITATIONS, REASONS FOR CAUTION
Results from this RCT may not be generalizable to other PRP preparations owing to heterogeneity and lack of standardization. The control groups did not undergo a sham ovarian injection, which would have been relevant had the results shown benefit of PRP injection. Only patients with POR were included in this study, and these results may not be generalizable to more severe diminution of ovarian reserve, as seen with POI.
WIDER IMPLICATIONS OF THE FINDINGS
The intraovarian PRP injection procedure does not improve mature oocyte yield or other parameters of IVF outcome in women less than 38 years old with an established IVF history of POR. The results from this study do not support the use of intraovarian PRP injection in this population.
STUDY FUNDING/COMPETING INTEREST(S)
Departmental funds were used and no external funding was requested for this study. ES is a consultant for and receives grant funding from the Foundation for Embryonic Competence. All other authors have no conflict of interest to declare.
TRIAL REGISTRATION NUMBER
Clinicaltrials.gov Registry Identifier: NCT04163640.
TRIAL REGISTRATION DATE
15 November 2019.
DATE OF FIRST PATIENT’S ENROLMENT
24 February 2020.
PubMed: 38725194
DOI: 10.1093/humrep/deae093 -
The EMBO Journal Jun 2024The 16-subunit Constitutive Centromere-associated Network (CCAN)-based inner kinetochore is well-known for connecting centromeric chromatin to the spindle-binding outer...
The 16-subunit Constitutive Centromere-associated Network (CCAN)-based inner kinetochore is well-known for connecting centromeric chromatin to the spindle-binding outer kinetochore. Here, we report a non-canonical role for the inner kinetochore in directly regulating sister-chromatid cohesion at centromeres. We provide biochemical, X-ray crystal structure, and intracellular ectopic localization evidence that the inner kinetochore directly binds cohesin, a ring-shaped multi-subunit complex that holds sister chromatids together from S-phase until anaphase onset. This interaction is mediated by binding of the 5-subunit CENP-OPQUR sub-complex of CCAN to the Scc1-SA2 sub-complex of cohesin. Mutation in the CENP-U subunit of the CENP-OPQUR complex that abolishes its binding to the composite interface between Scc1 and SA2 weakens centromeric cohesion, leading to premature separation of sister chromatids during delayed metaphase. We further show that CENP-U competes with the cohesin release factor Wapl for binding the interface of Scc1-SA2, and that the cohesion-protecting role for CENP-U can be bypassed by depleting Wapl. Taken together, this study reveals an inner kinetochore-bound pool of cohesin, which strengthens centromeric sister-chromatid cohesion to resist metaphase spindle pulling forces.
Topics: Kinetochores; Chromosomal Proteins, Non-Histone; Humans; Cell Cycle Proteins; Chromatids; Centromere; Cohesins; HeLa Cells; Protein Binding; Crystallography, X-Ray
PubMed: 38714893
DOI: 10.1038/s44318-024-00104-6