Clinical Application of In Vitro Maturation of Oocytes

In vitro maturation (IVM) is a technique used to induce immature oocytes collected in different periods of embryonic growth. The rates vary for immature oocytes collected from different clinical sources to potentially develop into embryos and achieve live birth. As an effective treatment method, IVM can be used to treat patients with polycystic ovary syndrome (PCOS), ovarian hyperresponsiveness, and hyporesponsiveness, as well as to preserve the fertility of cancer patients. This technology has been used worldwide for the birth of thousands of healthy babies. The improvement in clinical IVM technology mainly focuses on the IVM medium and the optimization of the culture environment and operation process. At present, with the improvement in the in vitro fertilization (IVF) efficiency and culture systems, a natural cycle or mild stimulation may be more suitable for women receiving IVF treatments. A new treatment option was proposed to combine natural cycle/mild stimulation IVF with IVM. In particular, the combination of mild stimulation IVF and IVM is not only expected to become a viable alternative to current standard treatments but may also become a potential option of first-line treatment

Lattice and Magnetic structures of PrFeAsO, PrFeAsO0.85F0.15 and PrFeAsO0.85

We use powder neutron diffraction to study the spin and lattice structures of polycrystalline samples of nonsuperconducting PrFeAsO and superconducting PrFeAsO0.85F0.15 and PrFeAsO0.85. We find that PrFeAsO exhibits an abrupt structural phase transitions at 153 K, followed by static long range antiferromagnetic order at 127 K. Both the structural distortion and magnetic order are identical to other rare-earth oxypnictides. Electron-doping the system with either Fluorine or oxygen deficiency suppresses the structural distortion and static long range antiferromagnetic order, therefore placing these materials into the same class of FeAs-based superconductors.Comment: 14 pages, 3 figures, 1 tabl

Transcriptional Regulation of an Evolutionary Conserved Intergenic Region of CDT2-INTS7

In the mammalian genome, a substantial number of gene pairs (approximately 10%) are arranged head-to-head on opposite strands within 1,000 base pairs, and separated by a bidirectional promoter(s) that generally drives the co-expression of both genes and results in functional coupling. The significance of unique genomic configuration remains elusive.Here we report on the identification of an intergenic region of non-homologous genes, CDT2, a regulator of DNA replication, and an integrator complex subunit 7 (INTS7), an interactor of the largest subunit of RNA polymerase II. The CDT2-INTS7 intergenic region is 246 and 245 base pairs long in human and mouse respectively and is evolutionary well-conserved among several mammalian species. By measuring the luciferase activity in A549 cells, the intergenic human sequence was shown to be able to drive the reporter gene expression in either direction and notably, among transcription factors E2F, E2F1 approximately E2F4, but not E2F5 and E2F6, this sequence clearly up-regulated the reporter gene expression exclusively in the direction of the CDT2 gene. In contrast, B-Myb, c-Myb, and p53 down-regulated the reporter gene expression in the transcriptional direction of the INTS7 gene. Overexpression of E2F1 by adenoviral-mediated gene transfer resulted in an increased CDT2, but not INTS7, mRNA level. Real-time polymerase transcription (RT-PCR) analyses of the expression pattern for CDT2 and INTS7 mRNA in human adult and fetal tissues and cell lines revealed that transcription of these two genes are asymmetrically regulated. Moreover, the abundance of mRNA between mouse and rat tissues was similar, but these patterns were quite different from the results obtained from human tissues.These findings add a unique example and help to understand the mechanistic insights into the regulation of gene expression through an evolutionary conserved intergenic region of the mammalian genome

Development and validation of a visualized prediction model for early miscarriage risk in patients undergoing IVF/ICSI procedures: a real-world multi-center study

BackgroundThis study focuses on the risk of early miscarriage in patients undergoing in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). These patients commonly experience heightened stress levels and may discontinue treatment due to emotional burdens associated with repeated failures. Despite the identification of numerous potential factors contributing to early miscarriage, there exists a research gap in integrating these factors into predictive models specifically for IVF/ICSI patients. The objective of this study is to develop a user-friendly nomogram that incorporates relevant risk factors to predict early miscarriage in IVF/ICSI patients. Through internal and external validation, the nomogram facilitates early identification of high-risk patients, supporting clinicians in making informed decisions.MethodsA retrospective analysis was conducted on 20,322 first cycles out of 31,307 for IVF/ICSI treatment at Sun Yat-sen Memorial Hospital between January 2011 and December 2020. After excluding ineligible cycles, 6,724 first fresh cycles were included and randomly divided into a training dataset (n = 4,516) and an internal validation dataset (n = 2,208). An external dataset (n = 1,179) from another hospital was used for validation. Logistic and LASSO regression models identified risk factors, and a multivariable logistic regression constructed the nomogram. Model performance was evaluated using AUC, calibration curves, and decision curve analysis (DCA).ResultsSignificant risk factors for early miscarriage were identified, including female age, BMI, number of spontaneous abortions, number of induced abortions and medical abortions, basal FSH levels, endometrial thickness on hCG day, and number of good quality embryos. The predictive nomogram demonstrated good fit and discriminatory power, with AUC values of 0.660, 0.640, and 0.615 for the training, internal validation, and external validation datasets, respectively. Calibration curves showed good consistency with actual outcomes, and DCA confirmed the clinical usefulness. Subgroup analysis revealed variations; for the elder subgroup (age ≥35 years), female age, basal FSH levels, and number of available embryos were significant risk factors, while for the younger subgroup (age <35 years), female age, BMI, number of spontaneous abortions, and number of good quality embryos were significant.ConclusionsOur study provides valuable insights into the impact factors of early miscarriage in both the general study population and specific age subgroups, offering practical recommendations for clinical practitioners. We have taken into account the significance of population differences and regional variations, ensuring the adaptability and relevance of our model across diverse populations. The user-friendly visualization of results and subgroup analysis further enhance the applicability and value of our research. These findings have significant implications for informed decision-making, allowing for individualized treatment strategies and the optimization of outcomes in IVF/ICSI patients

A Method for Structure–Activity Analysis of Quorum-Sensing Signaling Peptides from Naturally Transformable Streptococci

Many species of streptococci secrete and use a competence-stimulating peptide (CSP) to initiate quorum sensing for induction of genetic competence, bacteriocin production, and other activities. These signaling molecules are small, unmodified peptides that induce powerful strain-specific activity at nano-molar concentrations. This feature has provided an excellent opportunity to explore their structure–function relationships. However, CSP variants have also been identified in many species, and each specifically activates its cognate receptor. How such minor changes dramatically affect the specificity of these peptides remains unclear. Structure–activity analysis of these peptides may provide clues for understanding the specificity of signaling peptide–receptor interactions. Here, we use the Streptococcus mutans CSP as an example to describe methods of analyzing its structure–activity relationship. The methods described here may provide a platform for studying quorum-sensing signaling peptides of other naturally transformable streptococci

EWS/FLI Mediates Transcriptional Repression via NKX2.2 during Oncogenic Transformation in Ewing's Sarcoma

EWS/FLI is a master regulator of Ewing's sarcoma formation. Gene expression studies in A673 Ewing's sarcoma cells have demonstrated that EWS/FLI downregulates more genes than it upregulates, suggesting that EWS/FLI, and/or its targets, function as transcriptional repressors. One critical EWS/FLI target, NKX2.2, is a transcription factor that contains both transcriptional activation and transcriptional repression domains, raising the possibility that it mediates portions of the EWS/FLI transcriptional signature. We now report that microarray analysis demonstrated that the transcriptional profile of NKX2.2 consists solely of downregulated genes, and overlaps with the EWS/FLI downregulated signature, suggesting that NKX2.2 mediates oncogenic transformation via transcriptional repression. Structure-function analysis revealed that the DNA binding and repressor domains in NKX2.2 are required for oncogenesis in Ewing's sarcoma cells, while the transcriptional activation domain is completely dispensable. Furthermore, blockade of TLE or HDAC function, two protein families thought to mediate the repressive function of NKX2.2, inhibited the transformed phenotype and reversed the NKX2.2 transcriptional profile in Ewing's sarcoma cells. Whole genome localization studies (ChIP-chip) revealed that a significant portion of the NKX2.2-repressed gene expression signature was directly mediated by NKX2.2 binding. These data demonstrate that the transcriptional repressive function of NKX2.2 is necessary, and sufficient, for the oncogenic phenotype of Ewing's sarcoma, and suggest a therapeutic approach to this disease

Age-Specific Differences in Oncogenic Pathway Deregulation Seen in Human Breast Tumors

Purpose. To define the biology driving the aggressive nature of breast cancer arising in young women. Experimental Design. Among 784 patients with early stage breast cancer, using prospectively-defined, age-specific cohorts (young ≤45 years; older ≥65 years), 411 eligible patients (n = 200≤545 years; n = 211≥65 years) with clinically-annotated Affymetrix microarray data were identified. GSEA, signatures of oncogenic pathway deregulation and predictors of chemotherapy sensitivity were evaluated within the two age-defined cohorts. Results. In comparing deregulation of oncogenic pathways between age groups, a higher probability of P13K (p = 0.006) and Myc (p = 0.03) pathway deregulation was observed in breast tumors arising in younger women. When evaluating unique patterns of pathway deregulation, a low probability of Src and E2F deregulation in tumors of younger women, concurrent with a higher probability of P13K, Myc, and β-catenin, conferred a worse prognosis (HR = 4.15). In contrast, a higher probability of Src and E2F pathway activation in tumors of older women, with concurrent low probability of P13K, Myc and β-catenin deregulation, was associated with poorer outcome (HR = 2.7). In multivariate analyses, genomic clusters of pathway deregulation illustrate prognostic value. Conclusion. Results demonstrate that breast cancer arising in young women represents a distinct biologic entity characterized by unique patterns of deregulated signaling pathways that are prognostic, independent of currently available clinico-pathologic variables. These results should enable refinement of targeted treatment strategies in this clinically challenging situation. Copyright

Rapid Processing of Both Reward Probability and Reward Uncertainty in the Human Anterior Cingulate Cortex

Reward probability and uncertainty are two fundamental parameters of decision making. Whereas reward probability indicates the prospect of winning, reward uncertainty, measured as the variance of probability, indicates the degree of risk. Several lines of evidence have suggested that the anterior cingulate cortex (ACC) plays an important role in reward processing. What is lacking is a quantitative analysis of the encoding of reward probability and uncertainty in the human ACC. In this study, we addressed this issue by analyzing the feedback-related negativity (FRN), an event-related potential (ERP) component that reflects the ACC activity, in a simple gambling task in which reward probability and uncertainty were parametrically manipulated through predicting cues. Results showed that at the outcome evaluation phase, while both win and loss-related FRN amplitudes increased as the probability of win or loss decreased, only the win-related FRN was modulated by reward uncertainty. This study demonstrates the rapid encoding of reward probability and uncertainty in the human ACC and offers new insights into the functions of the ACC

Osteopontin induces growth of metastatic tumors in a preclinical model of non-small lung cancer

Osteopontin (OPN), also known as SPP1 (secreted phosphoprotein), is an integrin binding glyco-phosphoprotein produced by a variety of tissues. In cancer patients expression of OPN has been associated with poor prognosis in several tumor types including breast, lung, and colorectal cancers. Despite wide expression in tumor cells and stroma, there is limited evidence supporting role of OPN in tumor progression and metastasis. Using phage display technology we identified a high affinity anti-OPN monoclonal antibody (hereafter AOM1). The binding site for AOM1 was identified as SVVYGLRSKS sequence which is immediately adjacent to the RGD motif and also spans the thrombin cleavage site of the human OPN. AOM1 efficiently inhibited OPNa binding to recombinant integrin αvβ3 with an IC50 of 65 nM. Due to its unique binding site, AOM1 is capable of inhibiting OPN cleavage by thrombin which has been shown to produce an OPN fragment that is biologically more active than the full length OPN. Screening of human cell lines identified tumor cells with increased expression of OPN receptors (αvβ3 and CD44v6) such as mesothelioma, hepatocellular carcinoma, breast, and non-small cell lung adenocarcinoma (NSCLC). CD44v6 and αvβ3 were also found to be highly enriched in the monocyte, but not lymphocyte, subset of human peripheral blood mononuclear cells (hPBMCs). In vitro, OPNa induced migration of both tumor and hPBMCs in a transwell migration assay. AOM1 significantly blocked cell migration further validating its specificity for the ligand. OPN was found to be enriched in mouse plasma in a number of pre-clinical tumor model of non-small cell lung cancers. To assess the role of OPN in tumor growth and metastasis and to evaluate a potential therapeutic indication for AOM1, we employed a KrasG12D-LSLp53fl/fl subcutaneously implanted in vivo model of NSCLC which possesses a high capacity to metastasize into the lung. Our data indicated that treatment of tumor bearing mice with AOM1 as a single agent or in combination with Carboplatin significantly inhibited growth of large metastatic tumors in the lung further supporting a role for OPN in tumor metastasis and progression