Sleep and Vigilance

Abstract Background The global establishment of cancer registries has prompted a hunt for innovative medications that destroy cancer but not healthy cells. Researchers are currently searching marine environments for new anticancer drugs. Modern chemotherapy uses numerous compounds of aquatic origin. These substances exhibit cytotoxic characteristics through various methods, including DNA damage, apoptosis induction, and growth suppression of cancerous cells. The discovery and development of novel anticancer agents from abundant marine fish is generating increasing interest. The objective of this present study is to extract and assess the anti-proliferative effect of a crude muscle extract from marine pufferfish on human cancer cell lines. Methods The biochemical constituents, protein profile, and anticancer activities of the prepared crude muscle extract were analysed using standard methods on human cancer cell lines (HT-29, MDA-MB-231, A-549, and AGS). Results The biochemical contents, such as protein (7.19 ± 0.20 mg/mL), lipid (1.56 ± 0.14 mg/mL), and carbohydrate (1.19 ± 0.09 mg/mL), were quantitatively analysed. Native PAGE and SDS-PAGE qualitatively analysed the protein profiles, revealing distinct protein bands with molecular weights ranging from 220 to 14 kDa. The crude muscle extract was screened for its cytotoxicity (vero) and cell viability (HT-29, MDA-MB-231, A-549, and AGS) against human cancer cell lines by the MTT assay method. The nuclear morphological changes of the apoptotic cells were stained using propidium iodide, and the morphological changes associated with apoptosis were assessed using acridine orange/ethidium bromide (AO/EB) fluorescence staining. The intensity of the mitochondrial membrane potential of the treated cells was measured using the Rh-123 stain, and the results of the DNA fragmentation assay showed that the crude muscle extract-treated cells showed DNA damage, which is indicative of apoptosis. Conclusions This preliminary study supports that the crude muscle extract from milk spotted marine pufferfish, C. patoca has strong anticancer properties. This implies potential for the development of more effective anticancer drugs in future. Graphical abstract

Abstract Introduction Colorectal cancer (CRC) is a leading cause of cancer mortality in the USA, highlighting the urgent need for early detection. Methylated septin 9 (mSEPT9) is a promising tumor marker for CRC, offering high sensitivity for early stage detection, but its use in Asia is limited. Method We conducted a systematic review of empirical quantitative studies on mSEPT9 gene in cell-free DNA collected from plasma in CRC, following PRISMA and Cochrane guidelines, with searches yielding 1,340 records, of which 13 met our inclusion criteria. Quality was assessed using the STROBE checklist, ensuring robust data extraction and analysis. Result A total of 13 quantitative studies analyzed showed an overall positive detection rate of 66.98% for mSEPT9 as a diagnostic test for colorectal cancer, with the highest detection rate of 99.99% in Stage IV and the lowest at 51.21% in Stage I. Conclusion Our study demonstrates that mSEPT9 gene expression was a key biomarker for diagnosing and staging CRC, with higher detection rates in advanced stages, particularly Stage IV. Its role in critical signaling pathways highlights its potential for improving early detection and treatment planning in CRC patients.

Abstract Background Protein and glycoprotein antigens on the surface of red blood cells are encoded by highly polymorphic genes in human nuclear genomes. Allelic variants of the blood group genes are known to be distributed differently across populations. Consequently, they are widely used for ancestral analyses and for designing donor recruitment strategies. In Malaysia, blood group allele frequency population data are available for many population groups in Peninsular Malaysia, but not for those from Sabah and Sarawak of Borneo. Thus, this study contributes to a wider systematic collection of blood group data and expands the range of analyses. Aim This study determined the allele frequencies of nine clinically significant blood group systems (Kell, Kidd, Duffy, MNS, Dombrock, Colton, Lutheran, Cartwright and Vel) in the Bajau, Kadazan-Dusun, Murut and Melanau populations of Borneo. Results Many of the blood group systems examined here are polymorphic in the Bajau, Kadazan-Dusun, Murut and Melanau except for Kell, Colton, Lutheran, Cartwright and Vel. Population data mapping shows that these four populations are more closely related to other Austronesian populations, than other linguistically and historically unrelated population groups in Malaysia and elsewhere. Conclusions In this study, we have successfully typed variants that determine Kell, Kidd, Duffy, MNS, Dombrock, Colton, Lutheran, Cartwright and Vel blood group systems in the samples collected from unrelated Bajau, Kadazan-Dusun, Murut and Melanau individuals of Borneo. Many of these blood group systems were observed to be polymorphic in these populations. These findings are similar to those reported for other population groups in Peninsular Malaysia. Clinical complications that are more likely to arise in these people (gestation and transfusion alloimmunizations as well as risk to diseases) should be considered by health practitioners, healthcare providers and healthcare policy makers based on our observations. Graphical abstract

Abstract Background Colorectal cancer (CRC) is the third most frequent cancer and the second deadliest cancer, worldwide. Long noncoding RNAs (lncRNAs) have been introduced as crucial regulators of CRC. lncRNA feline leukemia virus subgroup C receptor 1 antisense RNA 1 (FLVCR1‑AS1) is suggested to play a significant role in the tumorigenesis of several cancers. The Wnt signaling pathway is the most deregulated pathway in CRC. Objective The present study aimed to investigate the underlying mechanism of function of FLVCR1-AS1 in CRC through FLVCR1-AS1/miR-381-3p/CTNNB1, LRP6, and FZD3 axis. Methods The expression levels of FLVCR1-AS1 were analyzed in colorectal cancer (CRC) tissues compared to adjacent normal tissues, as well as across various CRC cell lines. In HCT116 cells, FLVCR1-AS1 was knocked down, and the subsequent effects on the expression levels of FLVCR1-AS1, miR-381-3p, and three genes were measured using real-time PCR. Proliferation differences were assessed through an MTT assay, while cell death was evaluated using flow cytometry. Results The results confirmed that FLVCR1-AS1 was upregulated in CRC tissues compared to adjacent normal tissues. RT-qPCR validated that FLVCR1-AS1 has the most level of expression in HT29, HCT116, SW480, and Caco2; respectively. Knockdown of FLVCR1‑AS1 was significantly followed by attenuated viability of HCT116 cells; while resulted in enhancement of apoptosis and necrosis. Conclusion These findings support the idea that FLVCR1-AS1 may act as an oncogene in CRC, and targeting FLVCR1-AS1/miR-381-3p/CTNNB1, LRP6, and FZD3 axis may be introduced as a novel target for CRC therapy and diagnosis in the future.

Abstract Background Natural killer (NK) cells are crucial constituents of innate immunity, playing a vital role in the early defense against viral infections and cancer. Their antiviral capabilities stem from direct cytotoxic activity against infected cells and immunoregulatory functions that modulate adaptive immune responses. In the context of Coronavirus disease (COVID-19), NK cells contribute to controlling viral replication and limiting excessive inflammatory responses. Aims This review highlights the mechanisms underlying NK cell-mediated antiviral activity, focusing on their interactions with severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2). We discuss potential sources of NK cells for adoptive transfer therapy, critical factors influencing their therapeutic efficacy, and considerations for clinical translation in COVID-19 treatment. Methods Various NK cell sources for adoptive therapy are discussed, including peripheral blood, cord blood, and genetically engineered NK cells. Key factors influencing the therapeutic efficacy and safety of NK cell-based therapies are analyzed, including cytokine modulation, NK cell subset selection, and delivery methods. Conclusion Understanding the mechanisms underlying NK cell-mediated antiviral activity and the factors that influence the success of adoptive NK cell therapy can inform the design of more effective immunotherapies for COVID-19 patients. Strategies for improving NK cell function, such as genetic modification and optimizing therapy protocols, are highlighted to enhance therapeutic outcomes. Graphical Abstract NK cells neutralize SARS-CoV-2 by releasing cytotoxic granules, inhibiting replication, and modulating inflammation. They originate from peripheral blood, cord blood, and genetic engineering. Therapeutic strategies include adoptive transfer, cytokine modulation, and optimized delivery. Inflammation is controlled, reducing excessive immune responses.

Abstract Background Cerebral small vessel disease (CSVD) is a significant cause of ischemic stroke in all ethnic groups. Genetic determinants of CSVD are poorly understood and have rarely been explored. One proposed mechanism behind SVD is the breakdown of the blood–brain barrier (BBB). An intact barrier relies heavily on appropriate pericyte (PC) coverage of BBB endothelial cells. FOXF2 and NOTCH3 are two genes implicated in PC development and maintenance, thus influencing PC coverage of brain blood vessels. The aim of this study is to investigate whether FOXF2 rs41300825 and NOTCH3 rs1043994 variants can be considered as candidate loci for CSVD in Egyptian patients, marking the first study of its kind in Egypt. The present study included 186 ischemic stroke patients, selected from neurology and psychiatry department at Ain Shams University hospitals. Patients were categorized into: Group I (SVD patients; n = 114) and Group II (LVD patients; n = 72). Genotyping was conducted for FOXF2 rs41300825 (G/C) and NOTCH3 rs1043994 (A/G/T) using RFLP-PCR, with results confirmed through Sanger sequencing and multiple alignments of sequencing using the MEGA X program. Results Results showed that genotype analysis for FOXF2 rs41300825 displayed one band of 334 bp, indicating the GG genotype. The same was observed for NOTCH3 rs1043994 with one band of 665 bp, representing the GG genotype. Sanger sequencing and multiple alignments of sequencing using the MEGA X program confirmed the results. Conclusion Neither FOXF2 rs41300825 nor NOTCH3 rs1043994 were able to differentiate between patients at risk of developing SVD and those susceptible to LVD. Further studies with larger sample sizes are necessary.

Abstract Background The protein from suppressors of cytokine signaling (SOCS) family regulates immune response by modulating the signaling pathways of cytokines. SOCS2, a member of this family, plays an important role in regulating growth hormone receptors, the JAK-STAT pathway, energy homeostasis, and other biological processes. Multiple non-synonymous SNPs (nsSNPs) have been found in the SOCS2 gene, which could affect protein function. However, there is limited understanding of disease susceptibility and abnormal functioning associated with these mutated SOCS2 gene. Results In this study, we examined nsSNPs in the human SOCS2 gene to evaluate their effects on protein stability, structure, and function through in silico approaches. SIFT; PhD-SNP, PROVEAN, PMut, PANTHER, PolyPhen-2, SNPs&GO, I-Mutant 3.0, and MUpro were the bioinformatics tools used to forecast the most harmful SNPs. ConSurf found the eight nsSNPs (L71F, G102R, G51E, G47R, R96Q, Y49H, P155Q, and I171S) to be present in highly conserved region, thus affecting their protein stability. The Project HOPE analysis predicts the 3D structure of the eight respective mutated proteins and assesses their potential molecular impact on protein function and structure. After a series of analysis, three mutants (G47R, Y49H, and I171S) were subjected to molecular dynamic simulation, principal component analysis, and free energy landscape to understand their impact during the course of mutation. The STRING algorithm was used to predict protein–protein interactions. Finally, KM plotter analysis showed that deregulation of SOCS2 gene expression has a significant impact on the patients’ survival rate of different types of cancers. Conclusion Our study has identified the eight most high-risk SNPs of SOCS2 gene that may contribute to diseases development associated with growth hormone signaling, immune dysregulation, and other energy metabolism.

Abstract Background Several studies have linked the development of steroid-resistant nephrotic syndrome (SRNS) to variations in the multidrug resistance 1 gene (MDR1). However, disparities in findings have been noted among children of different ethnic origins. Objectives This study aimed to examine the relationship between MDR1 variants [rs944806668 (V1), rs1816042256 (V2), and rs1295312177 (V3)] and the risk of developing SRNS in Iraqi patients with idiopathic nephrotic syndrome (INS). Methods This case–control study included children with steroid-sensitive INS (n = 30) and SRNS (n = 30) from the Babylon Hospital for Maternity and Pediatrics. Sanger sequencing was used to determine the participants’ genotypes. Results Children with the V2 T/C genotype were more likely to develop SRNS [odds ratio (OR) and 95% confidence interval (95% CI): 10.8 (3.26–35.72), p < 0.001] than those with the wild genotype. However, no significant association with steroid response was observed for the V1 genotypes and alleles [OR (95% CI): 0.58 (0.18–1.91) and 0.63 (0.21–1.9), respectively; p > 0.05]. Moreover, children with the wild–variant–variant genotype combination of the V1, V2, and V3 variants exhibited a significantly higher risk of developing SRNS [OR (95% CI) 34 (4.91–235.61), p < 0.001] than those with the wild–wild–wild genotype combination. Conclusion These findings highlight the need to incorporate pharmacogenetic screening into the clinical management of SRNS. Children with nephrosis having the V2 T/C and V3 A/T genotypes, whether individually or in combination, are likely to resist prednisolone therapy, implying the need for alternative therapeutic approaches. Further studies are required to elucidate the potential implications of MDR1 variants for personalized drug therapy in children with INS.

Abstract Introduction Inherited retinal diseases (IRDs) are a clinically and genetically heterogenous group where the robust advancement of next-generation sequencing technologies has facilitated genotype-assisted diagnosis. Leber congenital amaurosis (LCA) is a severe form of inherited retinal dystrophy that causes congenital blindness or near-blindness with a global prevalence of 3 per 100,000 live births.It is characterized by a loss of vision at birth or within the first few years of life with overlapping phenotypes to many syndromic and non-syndromic IRDs. With India's rich genetic heterogeneity, WES is a valuable tool for uncovering novel gene mutations linked to LCA. This genetic diversity expands our understanding of the disease's spectrum in the Indian population. Methods In our previous study, 92 Indian LCA families were screened through targeted resequencing, and 80% of probands exhibited mutations in known genes. Hence, the remaining 20% probands with additional family members (n = 40) were subjected to whole-exome sequencing. An in-house standard bioinformatics pipeline was used for variant calling and annotation. Homology modeling (Modeller-9.23) and molecular simulation were performed on an identified SLC6A6 gene variant that has not yet been associated with LCA to investigate its potential pathogenicity. Results Disease-causing pathogenic variants were identified in 15/20 families (75%) across 11 genes with 33% variants being novel. Among the identified 17 variants in 15 families, 35% were missense, 29% nonsense, 29% frameshift and 6% splice variants. Segregation analysis, control screening and in silico predictions confirmed the variant’s pathogenicity. All variants were classified as pathogenic according to ACMG guidelines. Homology modeling and molecular simulation in the membrane system for the p.Pro82Leu mutant in SLC6A6 protein showed significant modification in helical characteristics around the TM2 helix in the mutant, which could potentially hinder the regular function and cause disruption in taurine transport across the membrane leading to the disease. Conclusion Taurine being an essential amino acid for photoreceptor development and maintenance, our study suggests that mutation identified in SLC6A6 gene may cause LCA. This is the first report of SLC6A6 gene association with LCA and also the first case report in the Indian population.

Abstract Objective Wolfram syndrome (WS) is a genetically disorder that affect on many organs, and neurodegenerative disorder. Although various clinical dysfunctions may have different onset times, they can collectively contribute to delays in the diagnosis of the disorder. To date, more than 200 pathogenic and likely pathogenic variant have been identified. In the present investigation, we evaluated three families with WS and reported a mutation in the WFS1. Methods This study, we have evaluated mutation in the WFS gene in three consanguineous families including three patients with a history of young-onset DM, progressive hearing loss and optic atrophy further neurological abnormalities. Results Sequencing results showed a novel homozygous stop-gain variant, c.1444A > T (p.K482X), and two previously reported mutations (c.2006A > G and c.2105G > A) in exon 8 of WFS1 gene. The variant interpretation was done according to the genetic guidelines. Finally, p.K482X was determined as a novel pathogen variant. Also, analysis showed that variants in parents were heterozygous. Conclusions The present survey, revealed a novel nonsense mutation in the wolframin protein, creates a frameshift which causes a premature stop codon truncating the protein in amino acid 482 residues. This mutation occurs in transmembrane domain and causes elimination of 46% of wolframin protein.

Abstract Background RP5-833A20.1, DYNLRB2-2, and APOA1 antisense are pivotal in atherosclerotic plaque pathogenesis. This study examined whether changes in these circulating lncRNAs could serve as biomarkers for high-risk ischemic stroke (IS) patients with intracranial atherosclerotic disease (ICAD). Methods Sixty-three IS patients, presenting within the first 24 h after stroke onset, and 60 controls were included in the study. The circulating levels of RP5-833A20.1, DYNLRB2-2, and APOA1 antisense in IS patients were assessed using real-time polymerase chain reaction (RT-PCR). Results Significant decreases in the circulating levels of DYNLRB2-2 and RP5-833A20.1 were observed in IS patients compared to controls (P < 0.05). However, no significant difference in APOA1 antisense levels was noted between the two groups. Subgroup analysis revealed higher RP5-833A20.1 expression in IS patients with lower National Institutes of Health Stroke Scale (NIHSS) scores (0–6) compared to those with higher scores (3.59 ± 0.783 vs. 1.05 ± 0.505, P = 0.006). After adjusting for relevant covariates, multiple logistic regression indicated an inverse association between RP5-833A20.1 and the risk of IS (adjusted OR = 0.846, P = 0.028). Linear regression analyses further demonstrated a negative correlation between RP5-833A20.1 expression and NIHSS (beta = − 0.398, P = 0.006), which was confirmed by a significant negative Spearman correlation (r = − 0.41, P = 0.0007). DYNLRB2-2 exhibited a non-significant negative relationship with NIHSS. Conclusion The findings suggest a significant decrease in the circulating levels of RP5-833A20.1 and DYNLRB2-2 in IS patients with ICAD, potentially indicating a protective effect against ischemic stroke. These lncRNAs hold promise as valuable biomarkers for identifying high-risk IS patients, emphasizing the need for further exploration and validation in larger cohorts to elucidate their roles in IS pathogenesis and clinical applications.

Abstract Background Colorectal carcinoma ranks as the second most prevalent neoplasm in females and the third most prevalent in males. Methods In this investigation, we examined the influence of alpha-pinene (ALP), classified as a monoterpene, on the proliferation of HT-29 human colorectal carcinoma cells via the PI3K/AKT signaling cascade. HT-29 cells were exposed to a range of ALP concentrations. The survival rate of the cells was assessed through the MTT assay; while, apoptotic events were evaluated using flow cytometry in conjunction with Annexin V/PI staining. Additionally, cell cycle dynamics were scrutinized through fluorescence-activated cell sorting. The expression levels of Bax, Bcl-2, Caspase-3, PI3K, and AKT at both the gene and protein levels were quantified employing real-time PCR and western blot techniques. Results ALP treatment led to a significant, dose-dependent decrease in cell viability, with prolonged exposure exacerbating cytotoxic effects. Apoptosis analysis showed that ALP significantly increased apoptotic cells after 24 h of treatment. Cell cycle analysis showed that ALP induces G2/M phase arrest and inhibits cell proliferation and DNA synthesis. Gene expression analysis revealed upregulation of pro-apoptotic genes Bax and caspase-3 and decreased anti-apoptotic gene Bcl-2. Furthermore, the administration of ALP led to a notable reduction in both the gene and protein expression levels of PI3K and Akt, suggesting a suppression of the PI3K/Akt signaling cascade. Conclusion ALP inhibits HT-29 colon cancer cell growth by reducing cell viability, inducing apoptosis, and blocking the PI3K/AKT pathway. These findings highlight ALP’s potential as a therapeutic candidate for colorectal cancer.

Abstract Background Nicotine metabolism significantly influences the levels of harmful nicotine metabolites in smokers. CYP2A13, a key enzyme in nicotine and xenobiotic metabolism, is implicated in tobacco smoke-related lung cancer. Aim This study investigated the association between CYP2A13 genetic polymorphism and nicotine metabolism in male smokers in Medan, Indonesia. Materials and methods This cross-sectional study included 66 male smokers (aged 20–65 years) who met pre-defined inclusion and exclusion criteria. Nicotine metabolite levels were quantified in urine samples using high-performance liquid chromatography (HPLC). CYP2A13 polymorphism was determined using polymerase chain reaction-restriction fragment length polymorphism (PCR–RFLP) analysis of venous blood samples. Logistic regression analysis (Epi Info-7) assessed the association between CYP2A13 genotype and nicotine metabolism. Results No significant association (p > 0.05) was found between CYP2A13 genotype and nicotine metabolism. The CC genotype was most prevalent. The majority of participants exhibited rapid nicotine metabolism. Conclusion Further research with larger sample sizes and diverse populations is needed to elucidate the relationship between CYP2A13 genetic polymorphism and nicotine metabolism.

Abstract Background Chromosomal aberrations are frequent in human embryos and can be identified during IVF by performing preimplantation genetic testing for aneuploidies (PGT-A). Therefore, this study presents a case report highlighting the application of embryonic cell-free DNA isolated from spent culture medium for preimplantation aneuploidy screening. Case presentation This case report elucidates the potential use of non-invasive preimplantation genetic testing for aneuploidy (PGT-A) as a valuable resource for couples facing infertility challenges. By using PGT-A, these couples can assess the genetic health of embryos before implantation, thereby increasing the chances of a successful pregnancy through assisted reproductive technologies. Conclusions PGT-A, an innovative technique using embryonic cell-free DNA isolated from a spent culture medium, offers a non-invasive and effective method for assessing chromosomal aberrations in embryos. It has the potential to enhance preimplantation genetic testing and support medically personalized embryo selection.

Abstract Background Type 2 Diabetes Mellitus (T2DM) is a significant global health concern characterised by insulin resistance and chronic hyperglycemia. Genetic factors, particularly variations in the CAPN-10 gene, have been implicated in T2DM susceptibility across diverse populations. Aim and objective This study aimed to conduct a meta-analysis to investigate the associations of single nucleotide polymorphisms (SNPs) in the CAPN-10 gene with T2DM among various populations, focusing specifically on Nigerian cohorts. Materials and methods A comprehensive literature search yielded 150 studies, from which 45 met inclusion criteria, encompassing approximately 25,000 individuals, including 10,000 diagnosed with T2DM. Statistical analyses assessed the association between CAPN-10 SNPs (UCSNP-43, UCSNP-19, UCSNP-63) and T2DM risk. Results A significant association was observed for UCSNP-43 (rs3792267) with T2DM (OR 1.50; 95% CI 1.28–1.75; p < 0.001), particularly in urban Nigerian populations. UCSNP-19 (rs3842570) also showed a moderate association (OR 1.35; 95% CI 1.10–1.66; p = 0.01), especially in South-West Nigeria. No significant association was found for UCSNP-63 (OR 1.15; 95% CI 0.90–1.45; p = 0.30). Conclusion The findings indicate that CAPN-10 SNPs, particularly UCSNP-43 and UCSNP-19, contribute to T2DM susceptibility in Nigerian populations, emphasising the importance of genetic screening for personalised interventions in diabetes management.