Search Results (286)

Hearing impairments (HIs) are clinically and genetically very heterogeneous. Finding the causative mutations in patients is frequently a challenge. We investigated two brothers affected by a sensorineural, moderate non-syndromic HI. Exome sequencing revealed that they carried the heterozygous c.812C>T (p.Ser271Leu) variant in GATA3. This gene encodes a transcription factor involved in embryonic development, its mutations causing the autosomal dominant HDR (hypoparathyroidism, deafness, and renal disease) syndrome. The variant affects a conserved residue within the proximal zinc-finger motif of GATA3. Sanger sequencing confirmed the presence of the variant in the two brothers, but it showed that surprisingly it was not carried by any of the parents. Segregation studies on 20 fully informative microsatellite markers in the family confirmed that the variant arose de novo. A benign SNP in the mother, close to the position of the variant, allowed us to determine that this was inherited from the father. Gene reporter functional assays supported the pathogenicity of the variant. Clinical reassessment of the two brothers did not disclose any additional abnormality. We conclude that mosaicism for this de novo mutation in the father’s germ line explains the pattern of inheritance in this family and that p.Ser271Leu is causing this unexpected phenotype of non-syndromic HI. Full article

Type II testicular germ cell tumors (TGCTs) are the most common solid malignancies in young men and are classified into seminomas and non-seminomatous subtypes. Seminomas are known for their highly pro-inflammatory tumor microenvironment (TME) with abundant immune cell infiltration. While previous work has demonstrated that the seminoma-derived cell line TCam-2 induces immune cell activation in co-culture and undergoes phenotypic changes itself, the underlying mechanisms remained unclear. To explore the role of direct cell–cell interaction and the effects mediated by soluble mediators such as cytokines, we conducted co-culture experiments of TCam-2 cells with purified human T cells or monocytes, including Transwell assays and treatments with IL-6, TNFα, or their respective blocking antibodies Tocilizumab and Adalimumab. In this way, we found that immune cell activation, indicated by enhanced secretion of pro-inflammatory cytokines and an upregulation of activation markers, strongly depended on direct physical contact between both cell types. Nonetheless, we also unveiled the role of soluble mediators in both immune cell activation and promoting a shift in TCam-2 cells from a seminoma-like phenotype to a more dedifferentiated phenotype, suggesting that cytokines critically shape the TME. These observations highlight the complexity of tumor–immune interactions in the seminoma microenvironment, offering new insight into immune-driven dynamics in TGCTs. Full article

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease affecting motor neurons with a phenotypic and genetic heterogeneity and elusive molecular mechanisms. With the present pilot study, we investigated different genetic mutations (C9orf72, TARDBP, and KIF5A) associated with ALS by generating induced pluripotent stem cells (iPSCs) from peripheral blood of ALS patients and healthy donors. iPSCs showed the typical morphology, expressed stem cell markers both at RNA (OCT4, SOX2, KLF4, and c-Myc) and protein (Oct4, Sox2, SSEA3, and Tra1-60) levels. Moreover, embryoid bodies expressing the three germ-layer markers and neurospheres expressing neural progenitor markers were generated. Importantly, the transcriptomic profiles of iPSCs and neurospheres were analyzed to highlight the differences between ALS patients and healthy controls. Interestingly, the differentially expressed genes (DEGs) shared across all ALS iPSCs are linked to extracellular matrix, highlighting its importance in ALS progression. In contrast, ALS neurospheres displayed widespread deficits in neuronal pathways, although these DEGs were varied among patients, reflecting the disease’s heterogeneity. Overall, we generated iPSC lines from ALS patients with diverse genetic backgrounds offering a tool for unravelling the intricate molecular landscape of ALS, paving the way for identifying key pathways implicated in pathogenesis and the disease’s phenotypic variability. Full article

Background/Objectives: Metastatic testicular germ cell tumor (mGCT) is a highly curable disease with first-line cisplatin-based combination chemotherapy. This study aims to evaluate the clinicopathological characteristics and survival outcomes of patients with metastatic testicular cancer in a nationwide multicenter cohort. Methods: This multicenter retrospective cohort study included 316 male patients diagnosed with mGCT who were treated with first-line cisplatin-based chemotherapy across 10 institutions in Turkey between 2011 and 2024. Clinical and pathological data, including International Germ Cell Cancer Collaborative Group (IGCCCG) risk classification, treatment details, and survival outcomes, were analyzed. Results: The median age of the cohort was 28 years, and 76.3% of patients were diagnosed with non-seminoma. According to IGCCCG risk stratification, 53.2% had good-risk, 25.3% intermediate-risk, and 21.5% poor-risk disease. Median follow-up was 38.4 months. Among patients with seminoma, the 5-year overall survival (OS) rate was 100% in the good-risk group and 87.5% in the intermediate-risk group. In patients with non-seminoma, 5-year OS rates were 96.6%, 86.9%, and 65.1% in the good-, intermediate-, and poor-risk groups, respectively. Among 125 patients who received salvage treatment, high-dose chemotherapy (HDCT) significantly improved survival in the International Prognostic Factors Study Group (IPFSG) very high-risk group (3-year OS: 55.0% vs. 16.3% with conventional-dose chemotherapy (CDCT), p = 0.007). Conclusions: This study provides the first large-scale nationwide dataset on mGCT outcomes in Turkey, demonstrating overall survival rates comparable to international cohorts. The findings emphasize the importance of a multidisciplinary approach, adherence to treatment guidelines, and optimal surgical interventions in improving patient outcomes. Full article

Chinese tongue sole (Cynoglossus semilaevis) is an important marine fish in China. It has sexual dimorphism. The weight and growth rate of female fish are much greater than those of male fish. However, high temperatures can induce sex reversal in genetic female fish (ZW) to phenotypic male fish; thus, identifying the genetic elements involved in temperature perception will provide the molecular basis for sex control. The heat shock transcription factor (hsf) is known as an important component of temperature sensing and mediates the heat shock response in fish such as Danio rerio; however, its function in C. semilaevis is unclear. In this study, five hsf genes (hsf1, hsf2, hsf4, hsf5a, and hsf5b) were identified in tongue sole and found to be expressed in the gonads at different developmental stages, peaking from 7M to 1Y. Gonadal in situ hybridization revealed that hsf gene signals were mainly localized in germ cells, e.g., sperm in the testis and all-stage oocytes in the ovary. Upon high-temperature stimulation, the expression of the hsf gene in the gonads increased gradually with increasing stimulation time, but different hsf genes presented different response patterns. After the RNA interference of hsf in the testis and ovarian cell lines, a series of sex-related genes, such as foxl2 and dmrt1, significantly changed. In vivo RNA interference had an effect on the female gonads and mainly affected neurl3 expression. On the basis of these data, we speculate that hsf responds to temperature stimulation and plays an important role in sex differentiation. This study helps elucidate the relationship between temperature sensing and sex differentiation in C. semilaevis. Full article

To address the growing consumer demands for improved fish meat quality, desirable morphological traits, and sustainable production practices, researchers have intensified efforts in the selective breeding and genetic improvement of carp (Cyprinus carpio) varieties. However, traditional breeding methods are often time-consuming and inefficient, which poses challenges to the sustainable development of the carp aquaculture industry. The establishment of germ stem cell lines offers a crucial tool for the study of germ cells, genetic improvement, and species conservation. In this study, we successfully established a spermatogonial stem cell line (YRSSCs) from Yellow River carp (Cyprinus carpio haematopterus) that can be cultured in vitro for the long term. We optimized the culture conditions to maintain their self-renewal and differentiation capabilities. The results demonstrated that YRSSCs have a diploid karyotype and can stably proliferate for over a year in L-15 medium supplemented with 5 mmol/L HEPES, 50 μmol/L β-mercaptoethanol, 15% FBS, 2 ng/mL bFGF, 2 ng/mL LIF, 1% carp serum, 800 IU/mL penicillin, 0.8 mg/mL streptomycin, 2 μg/mL amphotericin B, 1% zebrafish embryo extract, and 1% glutamine at 30 °C in the absence of CO₂. The cells exhibited a typical germ stem cell gene expression profile, with strong expression of the vasa, plzf-a, and Oct4-a genes. Additionally, this study found that YRSSCs possess the ability to differentiate in vitro and functionally colonize in vivo within recipient bodies. This research explored the establishment of YRSSCs and their differentiation potential both in vitro and in vivo, providing a novel strategy for the genetic improvement of aquaculture fish species through germ stem cell-based gene editing and transplantation technologies. Full article

The endoplasmic reticulum–Golgi apparatus system is an important organelle regulating male reproduction. USO1 vesicle transport factor (USO1), as an important molecule in this system, is a general vesicular transport factor and regulates various biological processes in vivo. However, the potential role of USO1 in mammalian testis development and spermatogenesis has not been investigated. We documented the presence of USO1 in mouse germ cells and its functional roles by generating Uso1-knockout germ cell lines. Uso1 depletion suppressed cell proliferation and growth while stimulating apoptosis in GC1 and GC2 cells. In addition, the Uso1 knockout blocked cell cycle progression and weakened DNA damage repair. Mechanistically, USO1 is associated with male reproduction by regulating the expression of genes involved in spermatogenesis, and we found evidence that USO1 is closely linked to centriolar satellites (CSs), which may play an important biological role. Overall, our findings reveal a vital role for USO1 in male fertility and offer a significant understanding of the functions of golgin proteins in reproductive biology. Full article

Inhibin (INH) plays a key role in the regulation of the reproductive performance of geese. It inhibits follicle-stimulating hormone (FSH) secretion from the anterior pituitary gland to regulate spermatogenesis. Immunization against INH in male geese leads to the production of antibodies to neutralize the INH activity that enhances testicular function and gonadotropin production. The objectives of the present research were to elaborate on the effects of inhibin (INH) immunization on testicular histology, plasma LH, pituitary PRL mRNA, and hypothalamic VIP mRNA expressions in Yangzhou ganders. A total of 60 birds were selected and divided into control (CON) and INH-immunized (INH-immunized) groups, having 30 in each group. In this experiment, the ganders were immunized with INH three times, and birds in the CON group were inoculated with bovine serum albumin (BSA). The analyzed data revealed that immunization against inhibin had no significant effects on improving the plasma concentration of LH hormone; however, significant effects were observed on the germ cell line, hypothalamic VIP mRNA, and pituitary PRL mRNA expressions. It is concluded that INH (INH) immunization is an effective tool to improve reproductive efficiency in Yangzhou ganders; however, INH immunization may harm pituitary PRL mRNA and hypothalamic mRNA expressions and LH plasma concentration. Seasonality played a vital impact on the hypothalamus–pituitary–gonadal (HPG) axis. Full article

Millimeter-waveband spectra of Venus from both the James Clerk Maxwell Telescope (JCMT) and the Atacama Large Millimeter/submillimeter Array (ALMA) seem to indicate there may be evidence (signal-to-noise ratio of about 15σ) of a phosphine absorption-line profile against the thermal background from deeper, hotter layers of the atmosphere. Phosphine is an important biomarker; e.g., the trace of phosphine in the Earth’s atmosphere is unequivocally associated with anthropogenic activity and microbial life (which produces this highly reducing gas even in an overall oxidizing environment). Motivated by the JCMT and ALMA tantalizing observations, we reexamine whether Venus could accommodate Earthly life. More concretely, we hypothesize that the microorganisms populating the Venusian atmosphere are not free floating but confined to the liquid environment inside cloud aerosols or droplets. Armed with this hypothesis, we generalize a study of airborne germ transmission to constrain the maximum size of droplets that could be floating in the Venusian atmosphere by demanding that their Stokes fallout times to reach moderately high temperatures are pronouncedly larger than the microbe’s replication time. We also comment on the effect of cosmic ray showers on the evolution of aerial microbial life. Full article

Mucolipidosis type II (ML II) is a rare and fatal disease of acid hydrolase trafficking. It is caused by pathogenic variants in the GNPTAB gene, leading to the absence of GlcNAc-1-phosphotransferase activity, an enzyme that catalyzes the first step in the formation of the mannose 6-phosphate (M6P) tag, essential for the trafficking of most lysosomal hydrolases. Without M6P, these do not reach the lysosome, which accumulates undegraded substrates. The lack of samples and adequate disease models limits the investigation into the pathophysiological mechanisms of the disease and potential therapies. Here, we report the generation and characterization of an ML II induced pluripotent stem cell (iPSC) line carrying the most frequent ML II pathogenic variant [NM_024312.5(GNPTAB):c.3503_3504del (p.Leu1168fs)]. Skin fibroblasts were successfully reprogrammed into iPSCs that express pluripotency markers, maintain a normal karyotype, and can differentiate into the three germ layers. Furthermore, ML II iPSCs showed a phenotype comparable to that of the somatic cells that originated them in terms of key ML II hallmarks: lower enzymatic activity of M6P-dependent hydrolases inside the cells but higher in conditioned media, and no differences in an M6P-independent hydrolase and accumulation of free cholesterol. Thus, ML II iPSCs constitute a novel model for ML II disease, with the inherent iPSC potential to become a valuable model for future studies on the pathogenic mechanisms and testing potential therapeutic approaches. Full article

Insects detect pathogens through their germ-line encoded pattern recognition receptors (PRRs). Among these, β-1,3-glucan recognition protein (βGRP) is a crucial PRR that specifically identifies pathogenic microorganisms and triggers innate immune signaling cascades. However, it remains unclear whether βGRP can detect viruses and protect the host from viral threats. In this study, using high-throughput sequencing technology, we observed a significant suppression of βGRP-3 in Bombyx mori during infection with the Bombyx mori cytoplasmic polyhedrosis virus (BmCPV). Moreover, overexpression of βGRP-3 in BmN cell lines resulted in a reduction of BmCPV proliferation, whereas knockdown of βGRP-3 in BmN cells promoted BmCPV proliferation. These findings suggest that the βGRP family functions not only as anti-bacterial, antifungal, and anti-yeast PRRs but also as protectors against various harmful viruses in insects. Full article

The testis-specific serine kinases (TSSKs) are post-meiotically expressed in testicular germ cells. Their testis-specific expression, together with their putative role in phosphorylation pathways, suggests that TSSKs have relevant roles in spermiogenesis, sperm function, or both. Independent Tssk3 and Tssk6 knockout mice, as well as the double Tssk1/Tssk2 KO males, are sterile. However, the double KO results are silent regarding the individual roles of TSSK1 and TSSK2. The aim of this study was to develop independent mutant mouse models of Tssk1 and Tssk2, using CRISPR/Cas9, to evaluate their independent roles in reproduction. Male heterozygous pups were used to establish one Tssk1 and two independent Tssk2 mutant lines. Natural mating mutant Tssk1 and Tssk2 homozygous males but not females were found to be sterile. Additionally, homozygous males have lower sperm numbers and decreased motility, and were infertile in vitro. Anti-TSSK2 antibodies were validated against Tssk2 mutants and used in Western blot and immunofluorescence experiments. TSSK2 is localized to the sperm head; importantly, it is present in the testes and sperm from Tssk1 mutant mice, confirming individual mutation. Our results indicate that both TSSK1 and TSSK2 are individually essential for male reproduction and support both kinases as suitable nonhormonal male contraceptive targets. Full article

Bovine pluripotent stem cells (PSCs) hold significant potential for diverse applications in agriculture, reproductive biotechnology, and biomedical research. However, challenges persist in establishing stable bovine PSC lines and understanding the mechanisms underlying their pluripotency maintenance. Here, we derived bovine embryonic stem cells (bESCs) from Holstein cattle embryos. These cells exhibited robust differentiation capacity into three germ layers in vitro and in vivo. Transcriptome analysis revealed distinct molecular profiles compared to primed-state bESCs. Notably, bESC proliferation ceased on methanol-treated feeder cells, in contrast to mouse ESCs (mESCs), which proliferated normally. Pathway analysis identified key signaling events critical for bESC survival and proliferation, highlighting species-specific regulatory mechanisms. Furthermore, the derived bESCs demonstrated chimerism capacity in early bovine embryos, underscoring their functional pluripotency. This work provides a foundation for advancing bovine embryology research and stem cell-based biotechnologies in livestock. Full article

Wheat germ oil (WGO), derived from the nutrient-dense germ of wheat kernels, is a functional bioactive product, known for its rich composition of essential fatty acids, sterols, tocopherols, and polyphenols. This study aimed to comprehensively profile the molecular and therapeutic properties of WGO, focusing on its antioxidant, cytotoxic, and anti-inflammatory activity. Using advanced analytical techniques such as gas chromatography-mass spectrometry (GC-MS), Fourier Transform Infrared (FTIR) spectroscopy, and fluorescence analysis, WGO was shown to contain high levels of linoleic acid (45.3%), squalene (2.52 g/100 g), and polyphenols. WGO displayed selective cytotoxicity, inhibiting cancer cells’ viability in melanoma, prostate, and colorectal cancer cell lines, but not normal cells, highlighting its chemoprevention potential. Furthermore, WGO significantly reduced LPS-induced nitric oxide and IL-6 production in macrophages, with effects plateauing at higher doses. The 3D fluorescence spectra showed a significant decrease in fluorescence intensity when human serum albumin interacted with the WGO polyphenol fraction, indicating a strong binding affinity and stable complex formation. These findings emphasize the nutritional and therapeutic potential of WGO as a natural bioactive agent, warranting further mechanistic investigation and broader applications in health and disease management. Full article

Methamphetamine (METH) use disorder (MUD) is a public health catastrophe. Herein, we used a METH self-administration model to assess behavioral responses to the dopamine receptor D1 (DRD1) antagonist, SCH23390. Differential gene expression was measured in the dorsal striatum after a 30-day withdrawal from METH. SCH23390 administration reduced METH taking in all animals. Shock Resistant (SR) rats showed greater incubation of METH seeking, which was correlated with increased Creb1, Cbp, and JunD mRNA expression. Cytoplasmic polyadenylation element binding protein 4 (Cpeb4) mRNA levels were increased in shock-sensitive (SS) rats. SS rats also showed increased protein levels for cleavage and polyadenylation specificity factor (CPSF) and germ line development 2 (GLD2) that are CPEB4-interacting proteins. Interestingly, GLD2-regulated GLUN2A mRNA and its protein showed increased expression in the shock-sensitive rats. Taken together, these observations identified CPEB4-regulated molecular mechanisms acting via NMDA GLUN2A receptors as potential targets for the treatment of METH use disorder. Full article