ObjectiveColorectal cancer is the third most common malignancy in the world and among the most frequent causes of cancer-related death. Our study aimed to evaluate the molecular profile of the patients diagnosed with colorectal carcinoma at Clinical Hospital Acibadem-Sistina in Skopje.Materials and methodsThis study is retrospective-prospective, conducted at the Department of histopathology and cytology, at Clinical Hospital Acibadem-Sistina in Skopje. Tissue samples from surgical material from 152 patients diagnosed with CRC were processed for molecular and immunohistochemical analysis. KRAS and BRAF mutations were analyzed, and MMR status was obtained. In 90 metastatic cases, evaluation of HER2 and PDL-1 expression was performed on a tissue microarray.ResultsAmong 152 analyzed patients diagnosed with colorectal carcinoma, the majority were males (98, 64.47%) compared to females (54, 35.53%). The mean age was 68.4±11.3 years; the median age was 70 years. KRAS/NRAS mutations were detected in 47(31%) of patients, BRAF mutations in 11(7%) patients, and mismatch repair gene deficiency (MMRd) was found in 15(10%) of patients. HER2 positive expression was present in 36(40%) of patients, and 17(19%) of patients showed PDL-1 expression. In the group of 17 PDL1-positive tumors, a cutoff of more than 1% positive tumor cells was detected in 10 cases, more than 10% tumor cells in 4 cases, and more than 50% tumor cells in 3 cases. From 36 HER2 positive cases, 32(32,5%) were with score 2+, and 4(4,4%) with score 3+.ConclusionsContinued research into molecular mechanisms and biomarkers holds the promise of further improving CRC outcomes through personalized and effective interventions.

The purpose of our study is to expedite cancer diagnosis through the development of software for rapid detection of hereditary breast cancer (BC) with negative BRCA1/2 on MATLAB, utilizing a fuzzy logic system with several variants of genes associated with BC. This system serves as a clinical decision-support tool, assisting in early classification and interpretation of genetic variants by combining clinical and genetic data. Clinical data were obtained from Erciyes University Faculty of Medicine Department of Medical Genetics and Uludağ University Faculty of Medicine Department of Medical Genetics. 488 individuals were studied. Only 90 of them were relevant to our investigation since their BRCA1/2 genes did not exhibit notable genetic mutations. We examined 16 distinct breast cancer risk factors and focused on mutations related to 18 hereditary BC genes. The collected data were integrated into the developed system, and various membership functions were given varying degrees of possibility, ranging from 0 to 1, depending on their participation in input clusters. After the system was trained on 90 cases and validated on six independent patients, its accuracy was assessed, yielding reliable results. Following the training phase, outcomes revealed the presence of two pathogenic variants at 0.92 (92%), two benign variants at 0.25 (25%), and two variants of unknown significance at 0.5 (50%). Given the high incidence of breast cancer, early prediction is paramount. Despite the emergence of fuzzy logic systems in medical applications, limited research akin to our study exists. The establishment of this artificial intelligence software holds promise for advancing the early detection of BC in future clinical applications.

Miscarriage, defined as the spontaneous loss of a fetus before viability, is the most common complication of pregnancy. Among its many causes, genetic factors are thought to play a significant role. One of the key signaling pathways involved in embryonic development is the Wnt/β-catenin pathway, which regulates critical processes such as embryonic cell migration, cell fate determination, proliferation, and differentiation. This pathway is also essential for early developmental events, including preimplantation development and blastocyst implantation. Although numerous animal studies have linked disruptions in Wnt signaling to pregnancy loss, limited data exist on its role in human miscarriage.In this study, we aimed to investigate the expression profiles of genes involved in the Wnt/β-catenin signaling pathway in placental tissues from a total of 23 miscarriage cases, including 15 with normal and 8 with abnormal fetal karyotypes.Our analysis revealed that GSK3B, WNT3A, WNT4, AXIN2, and APC were upregulated in the normal karyo-type group, while CTNNB1 (β-catenin) and WNT5A were downregulated. DVL1 expression showed no significant difference between the groups.These findings suggest that upregulation of GSK3B, AXIN2, and APC, together with downregulation of β-catenin, may lead to inhibition of the Wnt/β-catenin signaling pathway. Such disruption could impair key cellular processes—including proliferation, migration, and blastocyst implantation—that are essential for early pregnancy maintenance.

The discordance rate of the results between immunohistochemistry (IHC) and molecular microsatellite instability (MSI-PCR) tests, the most commonly used methods for qualitative deficient mismatch repair (dMMR) testing, is 1–10%, highlighting the need for a more precise testing strategy. The next-generation sequencing (NGS) offers a more sensitive and effective dMMR analysis (MSI-NGS), which also provides quantitative data. The aim of the study was to evaluate the qualitative and quantitative aspects of IHC and MSI-PCR testing compared to MSI-NGS in detecting dMMR in patients with Lynch Syndrome (LS)-associated and sporadic colorectal (CRC) and endometrial cancers (EC). Our results demonstrate both qualitative and quantitative discrepancies in the results obtained with different methods. Regarding qualitative differences, dMMR was inadequately interpreted only in LS when relying solely on IHC or MSI-PCR testing. This was primarily due to the specific mutational profile in our population, indicating the need for the implementation of a specific strategy that combines both methods. Concerning the quantitative differences, we detected great variability in the MSI levels, which was partly attributed to the tissue type or to the type of mutation in LS patients. Our results suggest that MSI-NGS level could be used as a potential surrogate marker for neoantigen levels and provide more precise predictive information for immunotherapy in patients with dMMR deficiency.

To characterize the prevalence of CYP2B6 and OPRM1 gene polymorphisms and evaluate their associations with serum methadone concentrations and dose requirements in Vietnamese patients undergoing methadone maintenance treatment (MMT). This cross-sectional study enrolled 200 patients with opioid dependence from multiple MMT clinics in Ninh Binh, Vietnam, between October 2023 and March 2024. Demographic, clinical, and behavioral data were collected using structured questionnaires. Blood samples were obtained for genotyping CYP2B6 (rs2279343 [785A>G], rs3745274 [516G>T], rs8192709 [64C>T]) and OPRM1 (A118G, rs1799971) using Sanger sequencing. Trough serum methadone concentrations were measured by validated liquid chromatography-tandem mass spectrometry (LC-MS/MS). Multivariable linear regression was performed to identify clinical and genetic factors associated with serum methadone concentrations, the concentration-to-dose ratio (CDR), and maintenance methadone dose. Among 200 participants (99% male, 96.9% Kinh ethnicity), the most common CYP2B6 diplotypes were *1/*4 (33.3%) and *4/*6 (23.7%), with the OPRM1 AG genotype present in 47.5%. Most patients (53.5%) had been on methadone treatment for over 3 years. In multivariable regression, the *1/*6 genotype was independently associated with lower maintenance dose (β = −26.33, 95% CI: −51.97, −0.69). The *2/*6 genotype was significantly associated with lower log serum methadone concentration (β = −2.485, 95% CI: −3.024, −1.947) and lower log CDR (β = −2.595, 95% CI: −3.145, −2.046). Most demographic, behavioral, and OPRM1 genotypes showed no significant associations. Specific CYP2B6 genotypes and SNPs significantly influence methadone pharmacokinetics in Vietnamese patients receiving MMT, although actual daily dose remains the most important determinant of serum levels. These findings highlight the potential role of genotype-guided methadone dosing in optimizing therapy for opioid dependence in Vietnamese populations.

IntroductionThe aim of this study was to compare the frequency of rs1205, rs1130864 and rs1800947 single nucleotide polymorphisms (SNPs) in the C-reactive protein (CRP) gene in rheumatoid arthritis (RA) patients and healthy controls. In addition, the aim was to investigate the association between these polymorphisms and serum CRP levels, Disease Activity Score-28 (DAS28) and clinical findings, especially extra-articular involvement such as interstitial lung disease.MethodsThis single-center study included 120 patients with RA and 100 healthy controls. DAS28-CRP was used to measure disease activity. Interstitial lung disease was confirmed by ground-glass opacities and interstitial thickening on chest computed tomography (CT) and other CT findings. Peripheral blood samples were collected in tubes with EDTA for genotyping. After DNA extraction, SNPs were analyzed using TaqMan probe-based polymerase chain reaction.ResultsAmong the three CRP SNPs (rs1130864, rs1205, rs1800947), a significant difference was observed in the distribution of rs1130864 between RA patients and controls. TT homozygous and CT heterozygous genotypes were significantly lower in the patient group (p = 0.002). Furthermore, the TT genotype of rs1130864 was significantly associated with lung involvement in patients with RA (p = 0.017). No significant association was found between SNPs and baseline CRP levels and DAS28 scores.ConclusionIn our study, we found a significant association between CRP rs1130864 gene polymorphism and lung involvement in RA patients. To the best of our knowledge, this is the first study to evaluate this association. However, further studies with larger cohorts are needed to confirm these findings.

IntroductionA group of primary bone tumors called chondrosarcomas are diverse and characterized by neoplastic tissue of hyaline cartilage. They are the second most typical primary osseous cancer. Recurrence is indicative of a poor prognosis because traditional chondrosarcomas are resistant to treatment. Our knowledge of the pathobiology of conventional chondrosarcomas has greatly expanded owing to recent findings in these tumors’ biology, genetics, and epigenetics studies. These findings also provide information on possible treatment targets. CHEK2 encodes a checkpoint kinase involved in DNA damage response and cell cycle regulation, and it is an important cancer susceptibility gene.MethodsThe Gene2 drug and DSEA assisted with in silico screening. The antitumor activities of the candidate drugs were extracted from DepMap via the PRISM viability assay on eight chondrosarcoma cell lines.Results and ConclusionAdvances in studies show promise for discovering potential targeted treatments for cancer. Bisacodyl is a targeted G protein-coupled receptor 35. G protein-coupled receptors are widely known targets for cancer treatment. Here, we showed that bisacodyl can be a potential therapeutic agent in chondrosarcoma cell lines.

A thorough functional evaluation of the immune system requires a method capable of measuring multiple cytokines to assess the adaptive immune response, as well as immunoglobulin isotypes and seroconversion to evaluate the humoral immune response following infections and vaccinations. Flow Cytometry Multiplex Bead Array (FCMBA) technology has been shown to be highly effective in detecting and quantifying numerous analytes simultaneously in a single reaction well. This technique is increasingly being used in immune profiling for cytokines in various disease contexts, as well as in adoptive cell transfer, immunotherapies, and more recently, in the characterization of humoral responses to newly developed COVID-19 vaccinations. FCMBA technology has shown advantages in assessing the depth of the humoral response mounted against COVID-19 vaccinations, compared to post-COVID infection status, and screening for the suitability of sera for use in COVID-19 convalescent plasma therapy. In this article, we present an overview of the FCMBA technology concept and its versatility, highlighting its ability to map many essential analytes and its superiority over conventional Enzyme Linked Immunosorbent Assay (ELISA) techniques. Furthermore, we discuss the current and potential clinical applications of this technology in deciphering COVID-19-triggered humoral immune responses, whether after contracting the infection or post-vaccination.

X-linked adrenoleukodystrophy (X-ALD) is a fatal neurodegenerative disorder caused by mutations in the adenosine triphosphate-binding cassette D1 (ABCD1) gene. In this study, we report the case of a Moroccan patient diagnosed with X-ALD due to a mutation in the ABCD1 gene. This diagnosis enabled a genetic counseling with presymptomatic analysis of the disease in the patient's brother, facilitating proactive management for the family. This work expands the clinical and genetic spectrum of XALD and underscores the critical role of presymptomatic testing in the management of neurodegenerative diseases.

Monoallelic KIF11 variants are associated with Microcephaly with or without Chorioretinopathy, Lymph-edema, or Impaired Intellectual Development (MCLMR) (OMIM:152950). This study presents seven patients from two distinct families, exhibiting extreme clinical heterogeneity, along with novel clinical findings and genetic variants. Patient 1 presented with learning difficulties, epilepsy, behavioral abnormalities, and various ocular anomalies, in addition to microcephaly and facial dysmorphism. Patient 2 was prenatally diagnosed with dorsal pedal edema, which was confirmed postnatally, alongside facial dysmorphic features and the need for craniosynostosis surgery at four months. Peripheral blood samples were obtained for DNA isolation, and Whole Exome Sequencing (WES) was performed for molecular analysis. Whole-exome sequencing (WES) revealed a de novo heterozygous frameshift variant in the KIF11 gene (NM_004523.4: c.2224_2225del; p. Asn742TyrfsTer6) in patient 1, and a novel heterozygous frameshift variant (NM_004523.4: c.2946dup; p. Leu983SerfsTer6) in the same gene in patient 2. Segregation analysis demonstrated that Patient 2's variant was inherited from the mother and was also present in the maternal grandmother, sister, and two aunts. Our findings highlight the broad clinical heterogeneity of MCLMR syndrome, as Patient 1 exhibited keratoconus, optic nerve hypoplasia, behavioral problems, and seizures, which are rarely reported. Additionally, Patient 2 represents the first documented patient of prenatal lymphedema detection in MCLMR syndrome. Furthermore, craniosynostosis, identified in two patients (P2 and P5), has not been previously described, suggesting a potential novel phenotypic feature. These findings expand the genotypic and phenotypic spectrum of KIF11-related disease.