This study evaluated squalene epoxidase (SQLE) mutations, and the antifungal susceptibility profile of Trichophyton species isolated from patients with recalcitrant dermatophytosis in Pakistan. The study was conducted at the Aga Khan University Hospital Laboratory, Karachi, between January 2023 and February 2024. Identification of the isolates was performed through sequencing of the internal transcribed spacer (ITS) region. Antifungal susceptibility testing for terbinafine, itraconazole, and voriconazole was performed using the broth microdilution method based on modified European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. Minimum inhibitory concentrations (MICs) were interpreted using EUCAST epidemiological cutoff values to classify isolates as wild-type or non-wild-type. DNA extraction was carried out using the QIAamp DNA Mini Kit, and ITS and SQLE gene regions were amplified and sequenced via Sanger sequencing. A total of 37 Trichophyton isolates were included, of which 17 (45.9%) were Trichophyton mentagrophytes, 16 (43.2%) were T. indotineae, 2 (5.40%) were T. rubrum, and 2 (5.40%) were T. interdigitale identified through ITS sequencing. Non-wild-type MICs to terbinafine were observed in 28 (75.7%) isolates, while eight isolates (21.6%) also showed non-wild-type MICs to itraconazole; no isolates demonstrated non-wild-type MICs to voriconazole. The most prevalent SQLE gene mutations were F397L (54%), Q408R (32.4%), and A448T (21.6%). This study represents the first report from Pakistan of antifungal resistance and SQLE gene mutations in Trichophyton strains from patients with recalcitrant dermatophytosis. The high prevalence of resistance underscores the urgent need for capacity building in antifungal susceptibility testing and highlights the importance of guiding appropriate treatment strategies to manage resistant dermatophytosis in the region effectively.

Gastrointestinal stromal tumors (GISTs) are the primary mesenchymal tumors in the gastrointestinal (GI) tract. Surgical treatments and tyrosine kinase inhibitors significantly improve patient outcomes. However, many patients develop resistance within 18-24 months, mainly due to mutations in the KIT and PDGFRA genes. Therefore, there is an urgent need for new biomarkers and therapeutic targets beyond KIT and PDGFRA to enhance diagnosis, monitor disease progression, and develop innovative treatment strategies. Gene expression data from GSE136755 were analyzed using R. Weighted gene co-expression network analysis identified co-expression modules, which were subsequently followed by GO/KEGG enrichment for functional insights. PPI networks were constructed using STRING, and hub genes were screened with CytoHubba. LASSO and ROC analyses evaluated the diagnostic value, while qRT-PCR and Western blot validated gene expression in GIST-T1 cells. Immune infiltration correlations were assessed using ssGSEA. Weighted gene co-expression network analysis identified 1323 genes in the MEblue module. Ten hub genes were recognized through PPI network analysis: CDK1, CCNB1, CCNA2, TOP2A, AURKA, AURKB, CDCA8, CHEK1, BUB1, and RAD51. Among these, 5 core signature genes were identified and validated through LASSO regression and ROC analysis, exhibiting strong diagnostic performance with AUCs ranging from 78.1% to 89.1%. Western blot and qRT-PCR tests validated these genes in GIST-T1 cells, and ssGSEA analysis indicated a significant relationship between these hub genes and immune cell infiltration. This study revealed a set of novel signature genes with high diagnostic value, offering promising targets for the diagnosis and treatment of GIST.

The role of cytokines in immune cell-mediated multiple myeloma and the identification of therapeutic targets.

Gastrointestinal Stromal Tumors (GISTs), derivatives of the Interstitial cells of Cajal, are the most common mesenchymal tumors of the gastrointestinal tract. Extra-gastrointestinal stromal tumors (EGISTs) are a subtype of GIST that represent less than 5% of all GISTs. EGIST arises either in the mesentery, omentum, or in the retroperitoneum, regardless of the gastrointestinal wall. Similar to GISTs, EGISTs are typically associated with mutations that result in gain of function in the KIT or PDGFRA receptor tyrosine kinase genes. Most EGISTs are found in the omentum and mesentery, and since they grow large in the abdominal cavity, most of them are large and palpable tumors. They are histologically defined as spindle or epithelioid cells, which are usually positive for CD117 (KIT) and DOG1. EGISTs are rare mesenchymal neoplasms that pose a significant clinical challenge due to their potential for aggression and a tendency to recur. We report a case of a high-risk, large mesenteric EGIST that had exhibited a complicated clinical course. The case went through an intraoperative tumor rupture, adjuvant treatment with imatinib, recurrence, and progression with increasing dose and second-line sunitinib treatment. EGISTs are more likely to have an aggressive clinical course and recurrence than GISTs of the same size and mitotic rate; therefore, aggressive treatment and long-term follow-up are needed. This case illustrates the impact of unfavorable prognostic factors, particularly tumor rupture and R1 resection, on the long-term prognosis of high-risk EGIST. It highlights the need for aggressive, multidisciplinary management, extended adjuvant therapy, and the challenges associated with managing sequential resistance in advanced EGIST treatment.

Relevance: Cutaneous melanoma is one of the most aggressive malignancies, in which mutations in the BRAF, NRAS, and KIT genes play a key role by activating the MAPK/ERK and RAS-RAF signaling pathways. In recent years, increasing attention has been given to the role of microRNAs (miRNAs) in regulating these signaling pathways. miRNAs can modulate the expression of oncogenes and tumor suppressor genes, influencing the growth, invasion, and therapy resistance of melanoma cells. Aim of the study: To identify the key “miRNA-gene” interaction pairs involved in melanoma pathogenesis and to evaluate their role in regulating major signaling cascades. Methods: Data from open-access databases NCBI and PubMed were used for analysis. Interactions between miRNAs and genes were investigated using the DIANA Tools bioinformatics platform, where interaction scores (Score) were calculated to reflect binding probability. Genes involved in the regulation of the cell cycle, apoptosis, and signal transduction were analyzed, as well as 30 of the most significant miRNAs associated with melanomagenesis. Results: Fifteen key genes (BRAF, NRAS, KRAS, KIT, NF1, MAP3K1, etc.) and 30 miRNAs modulating their expression were identified. The strongest interaction scores (Score ≥ 0.99) were found for the pairs BRAF-miR-5011-5p, KIT-miR-5011-5p, NRASmiR-4775, NF1-miR-3658, and HRH2-miR-4443. The study showed that miRNAs regulate the MAPK, PI3K/AKT, and RAS pathways, thereby affecting cell proliferation and apoptosis. Conclusion: The study identified new potential regulatory “miRNA-target gene” pairs associated with melanoma and demonstrated that post-transcriptional regulation plays a crucial role in melanoma pathogenesis. The findings suggest that specific miRNAs may serve as promising diagnostic biomarkers and therapeutic targets for melanoma.

To explore the correlation between ASXL1 gene mutation characteristics and clinical manifestations and prognosis in patients with myelodysplastic syndrome (MDS). The clinical date of 264 patients with MDS in Xuzhou Central Hospital, Southeast University from August 2010 to April 2024 was retrospectively analyzed. The patients were divided into ASXL1 wt group and ASXL1mut group according to the presence of ASXL1 gene mutation, and the correlation between gene mutation characteristics and clinical manifestations and prognosis was analyzed. Compared with ASXL1wt group, the ASXL1 mut group had a higher age of onset (P < 0.05), a higher proportion of males (P < 0.05), while the incidence of del(5q) was lower (P < 0.01). The mutation frequency of ASXL1 in MDS patients was 21.97%, and most of them were frameshift mutations. The p.Gly646fs was the most common amino acid variant, with a mutation frequency of 20.69%. The median overall survival (OS) and leukemia-free survival of patients with this sequence variant was 18.1 and 23.8 months, respectively, while in those without this sequence variant was 30 months and not reached, and the differences were statistically significant (P < 0.05). The results of multivariate analysis showed that the mutation of NRAS, WT1, KIT gene and the p.Gly646fs sequence mutation of ASXL1 gene were independent prognostic factors for OS in ASXL1mut patients. The median OS of ASXL1wt and ASXL1mut patients was 27.9(21.3-40.4) and 23.7(18.6-NA) months, respectively (P >0.05). Among 58 ASXL1mut patients, 5 cases (8.6%) transformed to acute leukemia, including 3 cases with RUNX1 mutation and 3 cases with TET2 mutation. Among 206 ASXL1wt patients, 28 cases (13.6%) transformed to acute leukemia. The difference in leukemia transformation rate between the two groups was not statistically significant (P >0.05). The efficacy of different treatment regimens was similar in the ASXL1mut group, while in the ASXL1wt group, patients receiving allogeneic hematopoietic stem cell transplantation had a significantly better prognosis than those receiving other treatment regimens (P < 0.001). The overall response rate to demethylation therapy was 68.7% and 67.6% in ASXL1mut and ASXL1wt group, respectively, and the difference between the two groups was not significant (P >0.05). The overall survival of MDS patients with ASXL1mut is poor. The patients with p.Gly646fs sequence mutation have a higher proportion of bone marrow blasts and a worse prognosis. There are no statistical differences in efficacy of different treatment strategies in ASXL1mut group. ASXL1 mutation shows no significant effect on the response of MDS to hypomethylating agent therapy.

IntroductionThe objective of this assessment was to determine the benefit of using a next-generation sequencing (NGS) gene panel for the clinical management of gastrointestinal stromal tumor (GIST) among patients in routine clinical practice. The aim was to assess the clinical utility of this procedure, the somatic molecular alterations of specific interest, and to define its role in the therapeutic care of patients with GIST.MethodsThe method used for this fast-track assessment were based on: (i) a critical analysis of systematic reviews, meta-analyses, and clinical practice guidelines identified by a systematic literature search; (ii) identification of the level of evidence of molecular alteration clinical actionability as set out by the European Society for Medical Oncology Scale, of targeted therapies included on the list of reimbursable drugs assessed by the French National Authority for Health, or drugs that have compassionate use authorizations issued by the French National Agency for Medicines and Health Products Safety; and (iii) stakeholder consultations and observations by public health institutions.ResultsAssessment of the evidence and data demonstrated that NGS gene panel testing has: (i) superior diagnostic performance for detecting KIT and PDGFRA molecular alterations, compared with Sanger sequencing; (ii) superior diagnostic performance for detecting NTRK1/2/3 fusion, compared with immunohistochemistry; and (iii) evidence of clinical utility of the targeted gene panel considering the benefits provided by targeted therapies.The composition of the NGS gene panel may be subject to change, in accordance with favorable assessments of new gene alterations. New assessments will be conducted in a dynamic manner in response to developments in scientific knowledge.ConclusionsThe French National Authority for Health deemed that funding the NGS gene panel for GIST was justified for: (i) KIT and PDGFRA genes in cases of locally advanced or metastatic GIST at an intermediate or high risk of recurrence, and in cases of suspected GIST when histology is inconclusive for diagnosis; and (ii) the NTRK1/2/3 gene in cases of refractory or relapsed locally advanced or metastatic pediatric wild-typeGIST.

Despite being one of the most iconic and immediately recognizable traits in domestic cattle, the variants underpinning the white-spotted coat pattern of Holstein-Friesian and related breeds remain uncharacterized. Here, we report two variants modulating these effects, comprising intronic and long-distance–acting regulatory variants of the MITF and KIT genes. We confirm causality through “Holsteinized” mouse models edited for these alleles and show that these variants are likely responsible for spotting traits in other bovine breeds. These effects include epistatic impacts on other bovine coat patterns, such as fine-scale speckling, “black socks,” and reversal of the otherwise dominant, “white-face” trait characteristic of Hereford cattle.

Clearance of RAS pathway mutations prior to allogeneic transplant in Acute Myeloid Leukemia are associated with favorable overall and leukemia free survival

Management of Mastocytosis and Mast Cell Activation in Children.

Resistance to 5α-reductase inhibitors (5ARIs) represents a significant therapeutic challenge in benign prostatic hyperplasia (BPH) clinical management. While the c-Kit-mediated signaling has been implicated in various pathological conditions, its role in BPH and 5ARI resistance remains undefined. Patient-derived organoids (PDOs) were established from BPH specimens and characterized through immunofluorescence, immunohistochemistry, and RT-qPCR analysis. Transcriptomic profiling was performed to identify differentially expressed genes between 5ARI-sensitive and resistant samples. The functional significance of c-Kit-mediated signaling was evaluated using selective inhibitor ISCK03. Further analysis identified cellular targets of c-Kit inhibition, and downstream signaling mechanisms were characterized through pathway analysis. RNA sequencing revealed differentially expressed genes between 5ARI-sensitive and resistant BPH PDOs, with significant enrichment in KIT and related genes. Enhanced c-Kit expression was confirmed in 5ARI-resistant specimens through multiple methodologies. Selective c-Kit inhibition with ISCK03 specifically suppressed 5ARI-resistant PDOs proliferation while sparing sensitive ones. Tests utilizing single-cell-derived organoids identified basal epithelial cells as primary targets of c-Kit inhibition. Mechanistic studies demonstrated that c-Kit maintains 5ARI resistance through the PI3K/AKT and Wnt/β-catenin signaling axis, with c-Kit inhibition significantly downregulating this pathway. c-Kit-mediated signaling is associated with 5ARI resistance in BPH, potentially through modulation of PI3K/AKT and Wnt/β-catenin pathways. These findings highlight c-Kit as a potential therapeutic target for overcoming 5ARI resistance.

IntroductionCameroon faced several waves of COVID-19 epidemics between 2020 and 2022. The epidemic peaks were characterized by variations in the number of positive cases and major variants. The aim of this study was to determine the frequency of COVID-19 in Cameroon during the different epidemic waves.Methodsnasopharyngeal samples were collected in different regions of Cameroon between 2020 and 2022. The Daan Gene kit (DaAn Gene, Guangzhou, Guangdong Province, China) was used to detect SARS-CoV-2 in the samples by RT-PCR assay. Excel 2013 software was used to record participants´ sociodemographic characteristics (age, sex, sampling date) and SARS-CoV-2 test results. Statistical analyses were performed using IBM SPSS version 25 software.Resultsfrom 16 March 2020 to 31 December 2022, 142,850 samples were tested. Participants ranged in age from 1 to 99 years with a M/F sex ratio of 1.32. Of the participants tested, 17,463 (12.2%) were positive for SARS-CoV-2. The SARS-CoV-2 detection rate decreased over time and was highest in 2020 (15.6%; 7255/46466) as opposed to 2021 (12.7%; 8859/69867) and 2022 (5.1%; 1349/26383). Four peaks of COVID-19 circulation were identified: May 2020, March 2021, September 2021 and December 2021. Risk factors for increased detection of SARS-CoV-2 were being older than 65 years and being from the Littoral region.Conclusionthe SARS-CoV-2 positivity rate in Cameroon decreased over the years, probably due to compliance with the barrier measures implemented by the Cameroonian government to reduce transmission rates.

Chronic lymphocytic leukemia (CLL) is characterized by a remarkably complex landscape. We performed a molecular characterization of liquid biopsies from 137 leukemia samples and 10 healthy subjects. Almost all patients express single-nucleotide polymorphism (SNP) mutations of the TP53 gene and the most common (n = 132 patients) was the Pro72Arg missense mutation. It was associated with significantly higher overall survival (p < 0.001) in untreated patients (n = 73) compared to treated patients (n = 58). However, this TP53 mutation is identified in both patients and healthy volunteers being likely a germinal mutation classified as SNP. In addition, a rare missense SNP mutation (Arg337Cys) of TP53 gene, correlated positively with clinical outcome (204 months; p = 0.00001) was also found. Its effects on p53 stability and function were subsequently evaluated. A missense mutation (Met541Leu) in the KIT gene was found in 18 patients and it was also significantly associated (p < 0.05) with a favorable outcome in untreated patients (n = 10) compared to treated ones (n = 8). We performed a comparative analysis among CLL patients classified into three prognostic groups (Binet A, B, and C). Survival analysis confirmed that Binet C patients had a significantly reduced survival compared to Binet A patients (p < 0.05).

Mast cell (MC) disorders result from inappropriate release of mediators and/or excessive accumulation of MCs, leading to symptoms of various organs and systems. Clonal MC disorders are defined by the presence of phenotypically aberrant and/or KIT-mutated MCs, and if aggregates of MCs are detectable, are designated as mastocytosis. Systemic mastocytosis (SM) affects mainly the bone marrow, with or without skin involvement. It is associated with the activating mutation D816V in the KIT gene. Other activating KIT gene variants are also observable in SM; activating KIT mutations are recognized as a minor diagnostic SM-criterion. We report a novel KIT variant in a patient with indolent SM, an in-frame deletion-insertion affecting amino acids D816 to N819 (D816_N819delinsll), creating an aliphatic pouch similar to that resulting from the D816V mutation, and leading to MC activation as suggested by the symptoms of the patient and the positivity for phosphorylated STAT5 in the clonal MCs.

Spermatogonial stem cells (SSCs) are essential for the continuous production of sperm and the maintenance of male fertility. Their selection, culture, and molecular characterization provide critical insights into spermatogenesis and potential therapeutic applications for male infertility. This study utilized CD49f-MACS and matrix selection techniques to isolate SSCs from mouse testicular samples. The molecular profile of the selected SSCs was analyzed through immunocytochemistry, gene ontology enrichment, weighted gene co-expression network analysis (WGCNA), and single-cell RNA sequencing (scRNA-seq). Additionally, protein-protein interaction (PPI) networks were constructed to identify key regulatory factors in SSC maintenance and differentiation. The selected SSCs exhibited a distinct molecular signature, with high expression of Dazl, Pou5f1 (Oct4), Gfra1, Nanog, and Kit. The Kit gene (c-kit) emerged as a crucial regulator of SSC differentiation, strongly associated with retinoic acid (RA)-mediated signaling pathways. Co-expression analysis revealed significant interactions between Kit, Nmyc, and other pluripotency-associated genes, highlighting its role in SSC development. Furthermore, single-cell RNA sequencing confirmed the dynamic expression of Kit during SSC differentiation and early meiosis initiation. Our findings underscore the pivotal role of Kit in spermatogenesis, reinforcing its potential as a therapeutic target for treating male infertility. The study also provides a comprehensive molecular framework for understanding SSC biology, with implications for regenerative medicine, fertility preservation, and in vitro gametogenesis. Further research integrating gene-editing technologies and in vivo models will be essential to explore the full therapeutic potential of SSC-based treatments.

Background/aimCystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CF is characterized by respiratory tract infections, pancreatic insufficiency, meconium ileus, intestinal obstruction, and male infertility. A genotype to phenotype correlation is difficult to establish because of the heterogeneity of disease severity. Even patients with the same CFTR mutation can have varying clinical severities. In recent years, studies have explored the role of microRNA (miRNA) expression in the regulation of respiratory diseases. However, no research has been conducted to date on miRNAs in siblings with the same CFTR mutation.Materials and methodsNasal cells of CF siblings from two families with discordant phenotype (n = 2 per family) were collected, and differentially expressed miRNAs were identified using miRNA arrays. Differentially expressed miRNAs and their target genes were determined using several bioinformatic databases and tools.ResultsmiR-449c-5p, miR-92b-3p, miR-34c-3p, miR-34c-5p, miR-6732-5p, and miR-4793-3p were differentially expressed in patients with severe disease compared to mild. CXCL1, CXCL2, DUSP1, GCLC, ICAM1, KIT, PRKAA2, and PTGS2 genes were identified as the target genes of candidate miRNAs (miR-34c-3p, miR-92b-3p, miR-449c-5p, miR-4793-3p). miRNA-mRNA interaction network analysis was performed and strong interaction was shown between miR-449c-5p target genes (CXCL1, CXCL2, PTGS2, ICAM1). CXCL1 expression decreased 5.28-fold in patients with severe disease compared to those with mild (p = 0.01).ConclusionOur results highlight the importance of miR-449c-5p interaction with CXCL1 and other target genes related to inflammation. Further studies should focus on the functional analysis of miR-449c-5p.

BackgroundResponse to trastuzumab-based neoadjuvant therapy in human epidermal growth factor receptor type 2 (HER2)-positive breast cancer is affected by multiple features of the tumor. Few studies have investigated epigenetic features in these patients. This study investigates whether changes in deoxyribonucleic acid (DNA) methylation patterns are linked to response to neoadjuvant therapy in HER2-positive breast cancer and aims to identify epigenetic markers of treatment resistance.Methods28 tumor samples were obtained from 20 HER2-positive breast cancer patients treated with neoadjuvant therapy: 12 from patients who achieved pathological complete response (pCR) before treatment, and 8 from patients who did not (non-pCR). For the non-pCR group, matched post-treatment samples were also collected, enabling paired pre- and post-treatment comparisons. After whole-genome methylation sequencing of all samples, the methylation differences between the pre-treatment pCR and non-pCR groups, as well as the methylation differences in non-pCR groups between pre-treatment and post-treatment samples were compared.ResultsBefore treatment, tumors in the non-pCR group showed slightly more hypomethylation events compared to the pCR group. After treatment, the same non-pCR tumors showed increased hypermethylation. Notably, immune-related pathways in these tumors were found to be hypermethylated, suggesting possible immune dysregulation. Methylation changes in the oncogenes MOS and RET were associated with potential resistance mechanisms. Additionally, four genes—KIT, LAD1, FAM110C, and DAPP1—were identified as candidate resistance markers based on their altered methylation patterns.ConclusionsThese findings highlight how DNA methylation changes may influence treatment outcomes in HER2-positive breast cancer and suggest novel epigenetic markers that could help predict or overcome therapy resistance.

Gastrointestinal stromal tumors (GISTs) are rare mesenchymal neoplasms of the digestive tract, most commonly originating in the stomach or small intestine, and driven by activating mutations in the KIT or PDGFRA genes. Liquid biopsy has emerged as a promising, minimally invasive technique to detect and monitor circulating tumor DNA (ctDNA), offering real-time insights into tumor dynamics and treatment response. Specifically, detecting KIT/PDGFRA mutations in ctDNA may aid in assessing prognosis, therapeutic response, and resistance. However, the clinical utility of this approach remains unclear. To address this, we conducted a systematic review and meta-analysis to evaluate the prognostic relevance of ctDNA mutations in GIST patients by comparing survival outcomes between those with KIT/PDGFRA mutations and those with wild-type profiles or no detectable ctDNA. A comprehensive systematic search was performed in the PubMed, Scopus, and Web of Science databases to identify studies evaluating overall survival (OS) at different time points in patients with GIST, stratified by ctDNA status (ctDNA-negative vs. ctDNA-positive). Hazard ratios (HRs) were extracted or calculated, and Kaplan-Meier curves were reconstructed using an adjusted Cox proportional hazards model, with 95% confidence intervals (CIs). A p-value < 0.05 was considered statistically significant. All statistical analyses were performed using RStudio software, version 4.2.3. This study included seven eligible studies comprising a total of 2024 histologically confirmed GIST patients, of whom 1610 were classified as ctDNA-positive and 414 had no detectable ctDNA mutations. OS at different time points was consistently more favorable in the ctDNA-negative group compared to the ctDNA-positive group (reference). The pooled hazard ratios (HR) were as follows: at 1year, HR 0.91 (95% CI: 0.89-0.93; p < 0.01; I2 = 0%); at 2years, HR 0.85 (95% CI: 0.83-0.88; p < 0.01; I2 = 20%); at 3years, HR 0.77 (95% CI: 0.74-0.81; p < 0.01; I2 = 28.2%); and at 5years, HR 0.63 (95% CI: 0.54-0.73; p < 0.01; I2 = 70.8%). At maximum follow-up (mean follow-up of 7.5months), OS showed a 49% reduction in survival in the ctDNA-positive group (HR 0.51; 95% CI: 0.40-0.64; p < 0.01; I2 = 79.9%). Additionally, in a pooled analysis of Kaplan-Meier data from patients with the KIT exon 11 (KIT11) mutation, the adjusted Cox proportional hazards model estimated an HR of 0.66 (95% CI: 0.49-0.89; p = 0.007), favoring the ctDNA-positive group. This meta-analysis highlights the potential of ctDNA as a prognostic biomarker in GIST, showing that its presence is consistently associated with poorer survival outcomes across mutational subtypes. These findings support the integration of ctDNA analysis into clinical practice as a minimally invasive tool for disease monitoring, contributing to more personalized and precise management of GIST patients.

The Swedish Mountain cattle and four other native Swedish cattle breeds show the phenotype of colour-sidedness. The causes of this phenotype are either a translocation and duplication from chromosome 6 to chromosome 29 (known as Cs29) including the KIT gene, or an additional translocation allele where part of Cs29 has been translocated back to chromosome 6 (Cs6). Besides the colour-sidedness, the Cs29 translocation has been associated with gonadal hypoplasia which can cause fertility problems. Despite breeding efforts during more than 80 years to reduce the prevalence of gonadal hypoplasia, there are still individuals with gonadal hypoplasia. We genotyped the Cs29 and Cs6 alleles in Swedish Mountain cattle and five other Swedish native breeds. Whole-genome sequence data of 30 cattle were analysed for the translocations, and 115 DNA samples were analysed using multiplex PCR. The current allele frequency of the Cs29 allele in Swedish mountain cattle was estimated to be 44%. The results indicate that the Cs29 translocation is also present in the four other native Swedish cattle breeds with colour-sidedness, and the Cs6 translocation in at least three of them.

ABSTRACT Background: Due to the complex histological and genomic nature of sarcomas, diagnosing and treating them has proven challenging. Delving into the genomic profiles and molecular markers linked to different sarcoma subtypes will aid in overcoming these obstacles and identifying new potential therapeutic targets. Objectives: The primary objective of this study was to investigate the genomic complexity of sarcoma, while the secondary objective was to identify potential therapeutic targets in the patients with sarcoma from India. Materials and Methods: This retrospective observational study was conducted from January 2020 to February 2024 at 4basecare Precision Health Pvt. Ltd., Bengaluru, India. We carried out comprehensive genomic profiling using gene panels or exome sequencing, including assessment of immunotherapy biomarkers (tumor mutation burden (TMB), microsatellite instability (MSI), programmed death-Ligand 1 (PD-L1)), in a cohort of 263 patients with sarcoma, categorized into 25 sarcoma types, for the present retrospective analysis. Results: We included 263 patients with sarcoma in our study and identified a diverse landscape of pathogenic variants across 138 genes, in 69.5% (183 patients) of the cohort. SNVs were prevalent in TP53 (25.1%; 66 patients), KIT (5.7%; 15 patients), PTEN (4.6%; 12 patients), and RB1 (4.6%; 12 patients), while CDK4 (5.2%; 17 patients) and MDM2 (5.7%; 15 patients) gene amplifications and SS18-SSX2 (1.1%; 3 patients), EWSR1-FLI1 (0.8%; 2 patients), and ASPSCR1-TFE3 (0.8%; 2 patients) gene fusions were recurrent. The majority of the patients harbored mutations affecting cell cycle control (39.2%; 103 patients), PI3K/AKT/MTOR (17.9%; 47 patients), and RAS/RAF/MAPK (14.8%; 39 patients) pathways. The average TMB was 7 mutations/mb, with 13.3% (35 patients) classified as TMB-H. Around 59.3% of the cohort (156 patients) harbored clinically actionable variants of therapeutic significance, including 8.7% of the cohort (23 patients) who were eligible for FDA/NCCN approved therapies. Conclusion: The findings emphasize the clinical usefulness of genomic profiling in guiding precision medicine for sarcoma treatment. Our research offers valuable insights into the genetic makeup of sarcomas, serving as a basis for devising efficient and precise diagnostic approaches and for planning preclinical and clinical studies to develop innovative treatment strategies.