The accumulation of chronic conditions may reflect shared biological susceptibility. We assessed whether the angiotensin-converting enzyme insertion/deletion polymorphism was associated with four prespecified, participant-reported, physician-diagnosed conditions. This multisite crosssectional study included 544 adults consecutively recruited from community settings across eight wilayas in Eastern Algeria. Multimorbidity was defined as at least two of the following conditions: hypertension, diabetes, thyroid dysfunction, and cardiac disease. Logistic regression was adjusted for age, sex, and tobacco use. Among the 544 participants, 84 (15.4%) had multimorbidity. Genotype was a significant contributor to the adjusted model (likelihood-ratio P=0.037). Compared with the deletion/deletion (DD) genotype, the insertion/deletion (ID) genotype was associated with higher odds of multimorbidity (adjusted odds ratio, 2.15; 95% confidence interval, 1.19-3.90; P=0.011), whereas the estimate for the insertion/insertion (II) genotype was imprecise and not statistically significant (adjusted odds ratio, 0.74; 95% confidence interval, 0.20-2.77; P=0.656). The ID genotype was associated with the presence of at least two conditions but not with exactly one condition, and no additive association with the D allele was detected. Genotype frequencies deviated from Hardy– Weinberg equilibrium (exact P=2.53 × 10^{-7} ), reflecting a heterozygote deficit. Bias analysis indicated that non-differential misclassification of heterozygotes as DD would attenuate the observed association. These findings suggest an association between the ID genotype and multimorbidity, although they should be interpreted cautiously and confirmed in independent studies.
Published acenocoumarol dose-prediction equations may perform differently across populations. We hypothesized that equations developed outside Algeria would show limited transportability. This study externally validated two pharmacogenetic equations and one matched clinical equation in 119 adults receiving stable acenocoumarol therapy at the University Hospital of Sétif, Algeria. Published coefficients were applied without refitting. Model-specific analyses included 109 participants for the Tong pharmacogenetic equation and 119 for the Tong clinical and Bulgarian pharmacogenetic equations, and direct comparisons used the common 109-participant subset. Performance was assessed using prediction error, prediction R², calibration, and the proportion of estimates within 20% of the observed dose, with 95% confidence intervals obtained from 5000 bootstrap resamples. Mean errors were -10.07, -7.44, and -8.21 mg/week for the Tong pharmacogenetic, Tong clinical, and Bulgarian equations, respectively. Corresponding mean absolute errors were 10.73, 8.96, and 10.48 mg/week, and 19.3%, 31.9%, and 26.9% of predictions were within 20% of the observed dose. All prediction R² values were negative, calibration was inadequate, and performance was poorest in patients requiring more than 21 mg/week. Sensitivity analyses supported the same interpretation. These equations substantially underpredicted stable dose and require formal recalibration and independent prospective validation before use in comparable Algerian patients.
In the biphasic life cycle of the smooth newts, the aquatic larvae transform into terrestrial adults, the metamorphs. The Greek smooth newt Lissotriton graecus is characterized by facultative paedomorphosis in which 2 adult morphs can coexist. Paedomorphs are reproducing adults that live in an aquatic habitat and retain some larval characteristics. In this study, histological and histochemical characteristics of the skin of the L. graecus in all morphs were investigated: larvae, paedomorphs, and metamorphs. The most remarkable finding is that skin development in paedomorphic L. graecus, whose skin is a mosaic of larval and metamorphic features, is partially uncoupled from sexual maturity. Leydig cells in the epidermis and numerous clear/vacuolated cells in the skin glands are common features with larval, while the relative thickness and keratinized surface of the epidermis are common features with adult skin. Our results provide new insights into this neglected field, given the gap in the existing literature on skin development in paedomorphic urodeles. The results of this study emphasized the importance of histological investigation of the possible uncoupling of sexual and somatic development in different organ systems to achieve a deeper understanding of the complex process of paedomorphosis.
The role of DNA repair gene polymorphisms in pancreatic cancer susceptibility remains insufficiently characterized, particularly in North African populations. This study investigated the association between the XPC Lys939Gln (rs2228001) polymorphism and pancreatic cancer risk in an Algerian cohort, alongside key metabolic risk factors. This exploratory hospital-based case-control study included 40 patients with histologically confirmed pancreatic cancer and 120 age- and sex-matched controls. Genotyping was performed using PCR-RFLP. Associations were evaluated using logistic regression models adjusted for demographic and metabolic variables. Given the limited sample size, the study was not powered to detect modest genetic effects. No statistically significant association was observed between XPC rs2228001 and pancreatic cancer risk under dominant, recessive, or allelic models (adjusted OR=1.22, 95% CI: 0.27-5.52; P=0.801). Type 2 diabetes mellitus was more frequent among cases and was associated with increased risk (OR=3.48, 95% CI: 1.24-9.77; P=0.018), although this finding should be interpreted with caution due to the exploratory design and limited sample size. The lower BMI observed in cases likely reflects disease-related weight loss. Under the constraints of this exploratory pilot investigation, no statistically detectable association was observed for the XPC rs2228001 variant. These findings are preliminary and should be considered hypothesis-generating. Larger, well-designed studies are required to clarify the interplay between genetic variation and metabolic factors in pancreatic cancer susceptibility.
Paracetamol administration at hepatotoxic doses represents a widely used experimental model to investigate drug-induced liver injury, causing significant hepatic damage through the generation of reactive metabolites that deplete glutathione and trigger oxidative stress, without necessarily causing mortality. This study evaluated the hepatotoxic effects of acute paracetamol administration and the hepatoprotective potential of Silybum marianum in Wistar rats. Biochemical markers (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and bilirubin) and liver histopathology were examined. Paracetamol induced marked histological liver damage, including hepatocellular necrosis and architectural disorganization, with only moderate and inconsistent changes in serum enzyme levels, indicating a dissociation between biochemical markers and structural hepatic injury. S. marianum treatment improved liver histology and partially stabilized biochemical alterations, supporting its hepatoprotective activity through antioxidant and membrane-stabilizing mechanisms. These results show that histopathological examination is a more sensitive indicator of acute hepatic injury than serum transaminases and support the therapeutic potential of S. marianum in paracetamol-induced hepatotoxicity.
Chronic wounds require topical formulations that combine favorable physicochemical properties with measurable biological activity. In this study, a bio-based, oil-in-water cream containing Teucrium polium methanolic extract was developed and optimized using a D-optimal design and a radial basis function (RBF) modeling approach. Three formulation variables, extract content (X1), cetyl alcohol (X2), and shea butter (X3), were evaluated in relation to two responses: spreadability (Y1) and wound closure on Day 7 (Y2). Predictive modeling was performed using an RBF network with leave-one-out cross-validation (LOOCV), which provided an internal estimate of predictive performance (R2 training = 0.98-0.99; R2 test = 0.95-0.96). Multi-response optimization identified an optimal formulation containing 8.0% extract, 1.5% cetyl alcohol, and 1.0% shea butter. Experimental confirmation yielded a spreadability of 54.8±0.9 cm2 and 84.1±1.2% wound closure on Day 7. In the confirmatory in vivo comparison, a vehicle control, a positive control (Madecassol 1%), and selected formulations were followed through Day 14. F6, F11, and F18 showed the most favorable macroscopic woundclosure profiles, whereas F2 displayed a more moderate effect. These findings support the use of a multivariate optimization strategy for the rational development of bio-based topical creams, while the biological interpretation remains limited to wound closure under the tested conditions.
The study evaluated the antioxidant and hepatoprotective potential of the n-butanol extract of Centaurea pungens (BECP) against carbon tetrachloride ($ \mathrm{CCl_4} $)-induced hepatotoxicity in Wistar rats. The safety of BECP was demonstrated through acute cytotoxicity tests and MTT assays. BECP showed in vitro radical scavenging activity. Pretreatment with BECP (100, 200, and 400 mg/kg) significantly alleviated liver markers in a dose-dependent manner, restoring antioxidant status. The results suggested that Centaurea pungens exerts strong hepatoprotective effects mediated by bioactive compounds identified by total phenolic content and LC-MS analysis, supporting its potential in liver disorder treatment.
Sphingosine-1-phosphate receptor 1 (S1PR1) modulators are an effective therapy for patients with relapsingremitting multiple sclerosis (RRMS), although considerable inter-individual variability in treatment response remains. This study aimed to investigate genetic variability within the coding regions of S1PR1 and to assess its potential association with treatment response and clinical disability measures, including the Expanded Disability Status Scale (EDSS) and Multiple Sclerosis Severity Score (MSSS), in patients with RRMS. Thirty-one RRMS patients from Serbia underwent sequencing of the 5/ regulatory region and coding exons of S1PR1 using an Applied Biosystems 3130 Genetic Analyzer. Three polymorphic variants were identified. None of the analyzed variants demonstrated a significant association with treatment response. Despite the small sample size, rs41287280 showed a significant association with disability measures; in a dominant inheritance model, carriers of the G allele had lower EDSS and MSSS values (P=0.01 and P=0.006, respectively) compared with the CC genotype. In multivariable analyses, the rs41287280 G allele was associated with lower disability, whereas the rs3737577 T allele was associated with higher disability compared to the wild-type GG genotype after adjustment. These results provide preliminary evidence suggesting that S1PR1 genetic variation may be related to disease pathogenesis in RRMS, warranting further confirmation in larger, independent cohorts and, potentially, functional investigation.
Reproductive traits are important taxonomic characters in section Polygonum. However, the taxonomic value of this character remains uncertain because of morphological variability and overlap among taxa. The goal of this research was to evaluate the discriminatory power of a comparative analysis of the quantitative and qualitative morphological characters of the bracts, perianth, and fruit in selected representatives of sect. Polygonum. The sample included seven taxa from 54 populations inhabiting the Balkan Peninsula and the Pannonian and Wallachian plains, and 23 quantitative and 9 qualitative morphological characters were analyzed. The results revealed a high degree of similarity between certain taxa, particularly P. aviculare and P. arenastrum, as well as P. arenarium subsp. arenarium and P. arenarium subsp. pulchellum. Almost all qualitative characters, with the exception of pericarp ornamentation, were found to be unstable at the taxon level, calling into question their previously emphasized taxonomic significance. Although newly analyzed quantitative characters provided additional information, they did not enable complete separation of the investigated taxa. Overall, the results indicate that morphology alone has limited resolving power within sect. Polygonum and highlight the need for an integrative taxonomic approach combining morphological, molecular, and ecological data.
We hypothesized that VKORC1 genetic variation contributes to stable acenocoumarol maintenance-dose requirements in Algerian patients. This prospective single-center study included 119 of 321 adults assessed for eligibility in Sétif between November 2017 and September 2019. Eligible patients had received acenocoumarol for at least six months, maintained an indication-specific international normalized ratio within the therapeutic range for at least three months, and achieved a time in therapeutic range >60% according to the Rosendaal method. The stable maintenance dose was defined as the mean prescribed daily dose recorded during visits with therapeutic international normalized ratio values. VKORC1 rs9923231, rs7294, and rs17708472 were genotyped using polymerase chain reaction– restriction fragment length polymorphism. The mean maintenance dose was 3.08±1.60 mg/day. For rs9923231, the mean maintenance doses were 4.27, 3.42, and 2.11 mg/day in GG, GA, and AA genotype carriers, respectively. Compared with GG carriers, adjusted dose ratios were 0.87 (95% confidence interval [CI], 0.71-1.07) for GA and 0.53 (95% CI, 0.43-0.66) for AA (overall P < 0.001). The rs7294 variant was associated with a higher maintenance dose (dose ratio 1.18, 95% CI 1.05-1.34; Holm-adjusted P=0.016), whereas rs17708472 showed no independent association. These findings demonstrate that VKORC1 rs9923231 is a major determinant of stable acenocoumarol maintenance-dose requirements in this Algerian cohort, with the greatest dose reduction observed in AA homozygotes.
Oxygen deprivation, resulting from various pathological conditions, is a major cause of brain dysfunction throughout development and adulthood. Despite extensive preclinical research, much of it conducted in animal models, proposed neuroprotective treatments have largely failed in clinical trials. In the present study, using a human cell-based in vitro model of short-term hypoxic injury, we identified molecular alterations associated with impaired neural fate determination, ultimately resulting in a significant reduction in neuronal yield. RNA sequencing and functional enrichment analyses revealed that exposure to cobalt chloride, a hypoxia-mimetic agent, induced a robust transcriptome response in NT2/D1 embryonal carcinoma stem cells, including metabolic dysfunction and increased expression of regulatory genes involved in multiple developmental processes and survival. More than 200 genes associated with neurogenesis and over 100 genes associated with synaptic signaling were significantly downregulated in early neural progenitor cells derived from CoCl₂-treated NT2/D1 cells. These findings were consistent with the observed morphological and immunocytochemical characteristics of the cells. Importantly, our study demonstrates that cobalt chloride selectively modulates the expression of genes involved in the differentiation of various cell populations, underscoring the need for further research to elucidate how this crucial cellular response to hypoxia shapes brain development and cognition.
This study investigates the ecological and morphological traits of invasive pumpkinseed (Lepomis gibbosus) in five lowland reservoirs in Serbia. A total of 434 specimens were collected in October 2020 and analyzed for population structure and growth parameters, body shape, condition, and diet. The population consisted of seven age classes, with three distinct age structures identified across the analyzed reservoirs. The highest growth increment occurred during the first year of life. Compared to other European populations, this cohort exhibited the slowest overall growth, characterized by the lowest k values. However, the specimens maintained a robust body condition (K>1) and displayed positive allometric growth (b>3). The autumn diet included 14 prey categories, with Chironomidae larvae being the most prominent. Mean dietary similarity between reservoirs was 75.7%, indicating significant dietary niche overlap. Niche breadth analysis showed greater dietary specialization in Topola and Kudre & ccaron; 2, and lower specialization in Jatagan, where prey diversity was highest. These findings offer valuable insights into the adaptive potential and ecological impact of this invasive species and can support future management and monitoring strategies.
Biosynthesis of metal nanoparticles using plant extracts is considered an eco-friendly approach with promising biomedical applications; however, data on their safety, particularly genotoxicity, remain limited. This study evaluated the phenolic composition and genotoxic potential of aqueous extracts from the root (SPR) and aerial parts (SPA) of Salvia pratensis L. and their corresponding biogenic silver nanoparticles (SPR-AgNPs and SPA-AgNPs). Spectrophotometric methods were used to quantify total phenolic and flavonoid contents, and LC-MS analysis was employed to characterize the phenolic profiles. The genotoxic potential of the extracts and AgNPs was evaluated in human lymphocytes via the comet assay, with treatments applied at four concentrations ranging from 25 to 100 & micro;g/mL, either independently or in combination with hydrogen peroxide (H2O2). Both extracts exhibited high phenolic content, with lower flavonoid levels in SPR. LC-MS analysis identified rosmarinic acid as the predominant compound in SPA and salvianolic acid K in SPR. Both extracts and AgNPs increased DNA damage in lymphocytes. In contrast, only the SPA extract lowered H2O2-induced DNA damage, demonstrating a pronounced protective effect. These findings indicate potential genotoxic risks associated with S. pratensis extracts and their AgNPs, while also highlighting selective protective properties depending on the plant part used.
This study evaluated the fumigant toxicity of Thymus munbyanus subsp. coloratus essential oil against adult Ephestia kuehniella (Zeller). To determine sublethal effects, we integrated biochemical biomarkers with assessments of reproductive performance and locomotor activity. Adults were exposed via fumigation to concentrations between 0.05 and 0.8 mu L/mL of air, with lethal concentrations calculated after 24 h of exposure. The essential oil showed strong fumigant activity, with LC25 and LC50 values of 0.153 and 0.26 mu L/mL of air, respectively. Sublethal exposure at these concentrations was associated with changes in biomarker responses in adults, including increased glutathione S-transferase (GST) and catalase (CAT) activities, along with decreased acetylcholinesterase (AChE) activity. In addition, fumigation affected adult reproductive performance by reducing oviposition, as reflected by a decrease in the number of eggs laid and hatched. Locomotor activity was also impaired, with reductions in distance traveled, movement duration, and movement speed. Overall, these findings suggest that T. munbyanus essential oil has potential as a botanical fumigant against E. kuehniella adults.
Pancreatic cancer risk was investigated in relation to the angiotensin-I-converting enzyme (ACE) insertion/deletion (I/D) polymorphism and its interaction with type 2 diabetes mellitus in an Eastern Algerian population througha hospital-based case-control study. The research involved 35 patients with confirmed pancreatic cancer and 140 matchedhealthy controls. Genotyping was performed to determine ACE I/D profiles, and statistical analysis was used to quantifyrisk associations. Type 2 diabetes mellitus was common, affecting more than half of the patients. The heterozygous IDgenotype was associated with a 2.5-fold increased cancer risk, and this association was significant only among individualswith diabetes. Multivariable analysis suggested the ID genotype as an independent risk factor, with an approximately 8-foldhigher risk after adjustments. This preliminary study suggests the ACE I/D polymorphism may influence pancreatic cancerrisk, with type 2 diabetes acting as a critical effect modifier and identifying a potential high-risk subgroup for targetedsurveillance. These exploratory findings warrant validation in larger, population-based cohorts.
Marine fungi are known for their ability to produce bioactive compounds with pharmaceutical potential. However, their growth in diverse and often extreme habitats makes laboratory cultivation difficult, thereby limiting research progress. The algicolous endophyte fungus Aspergillus terreus strain BAWK-F6 was isolated from brown seaweed Dictyota dichotoma and identified through molecular analyses. The fungal extract was evaluated for a range of biological activities. Molecular identification revealed a high genetic similarity with other A. terreus strains, and chemical analysis using gas chromatography mass spectrometry (GC-MS) identified 28 compounds. The extract demonstrated antioxidant activity, with IC50 values ranging from 1.98 to 2.32 mg/mL. Antimicrobial assays demonstrated that the extract effectively inhibited a broad spectrum of Gram-negative pathogens, with minimum inhibitory concentrations (MIC) ranging from 0.625 to 1.25 mg/mL. The extract also exhibited potent antibiofilm activity, especially against Staphylococcus aureus. It exhibited anti-inflammatory activity by inhibiting protein denaturation and stabilizing red blood cell membranes, as well as antiurolithic potential by suppressing the formation of calcium phosphate crystals. These findings highlight the therapeutic promise of Aspergillus terreus BAWK-F6 as a source of bioactive compounds with potential applications in pharmacology and biotechnology.
Tuberculosis is a global public health concern, and its treatment is frequently associated with hepatotoxic sideeffects. Phytotherapy represents a promising complementary approach. This study aimed to characterize the phenolic profileof the n-butanol extract of Crotalaria vialattei (BCV), evaluate its antioxidant capacity, and assess its hepatoprotectiveeffects against liver damage induced by a fixed-dose antituberculosis drug combination containing rifampin, isoniazid, andpyrazinamide (RHZ). Phytochemical analysis revealed a phenolic-rich extract, identifying 17 polyphenolic compounds.BCV exhibited measurable antioxidant activity in vitro. In vivo, oral administration of RHZ (rifampin 150 mg/kg, isoniazid75 mg/kg, and pyrazinamide 400 mg/kg) induced marked alterations in hepatic biochemical markers, lipid profile, oxidativestatus, and liver histoarchitecture. BCV treatment significantly attenuated these changes by improving liver enzyme activities,restoring oxidative balance, and preserving liver architecture. Overall, the BCV extract demonstrates antioxidant-associatedhepatoprotective potential against RHZ-induced liver injury and may represent a promising complementary strategy toreduce antituberculosis drug-related hepatotoxicity.
The current research aimed to investigate the potential effectiveness of single and combined supplementation with troxerutin (Trx) and selenium (Se) against cypermethrin (Cyp)-induced neurotoxicity and oxidative stress in mice. Thirty mice were randomly divided into five groups. The control group received 0.9% normal saline orally; the Cyp group received Cyp (5 mg/kg) orally; the Cyp+Trx group received Trx (150 mg/kg) in combination with Cyp; the Cyp+Se group received Se (25 & micro;g/kg) with Cyp; and the Cyp+Trx+Se group received Trx (150 mg/kg) and Se (25 & micro;g/kg) together with Cyp, all administered orally for 28 days. The individual and combined supplementation of Trx and Se significantly improved motor impairment, memory function, hepatic and renal health markers, and reduced anxiety levels (inside time/time in centar zone) in Cyp-exposed mice. Moreover, all supplementations reduced the Cyp-induced oxidative stress, but the SOD activity was increased in the serum and brain by the supplementation of Trx and Se, respectively. Liver and kidney SOD activities were improved with all the supplementations. Combined supplementation of Trx and Se enhanced brain acetylcholinesterase activity compared to the Cyp group. These findings suggest that both single and combined supplementation with Trx and Se confer neuroprotection against pesticide-induced oxidative and behavioral alterations; however, brain acetylcholinesterase activity and renal health markers were improved only by the combined Trx and Se supplementation.
The genetic variant IL2RA rs2104286 (A>G) is strongly associated with multiple sclerosis (MS) risk. It modulates IL-2 signaling, thereby influencing the expression of lymphocyte function-related genes ORMDL3 and IKZF3 that are implicated in multiple sclerosis pathogenesis. This study aimed to investigate the association of rs2104286 with ORMDL3 and IKZF3 mRNA expression in peripheral blood mononuclear cells (PBMCs) of relapsing-remitting multiple sclerosis (RRMS) patients and controls. The study included 66 RRMS patients and 45 healthy controls from the Serbian population. Genotyping and gene expression analysis were performed using qPCR. In RRMS patients, increased ORMDL3 and IKZF3 mRNA expression was associated with the MS-protective G allele (ORMDL3: P = 0.014, 1.35-fold increase; IKZF3: P = 0.023, 1.38-fold increase). In controls, decreased expression of IKZF3 was associated with the G allele (P=0.038, 0.72-fold change). Sex, disease phase, interferon (3 therapy, and the ORMDL3 and IKZF3 expression quantitative trait locus (rs12946510) did not influence the association between target gene expression and rs2104286. The overall results suggest a disease-specific association between the IL2RA rs2104286 variant and ORMDL3 and IKZF3 expression. Our findings support the notion that the rs2104286 variant may modulate IL-2 signaling in PBMCs, thereby influencing the expression of the immune-relevant genes ORMDL3 and IKZF3.
We report the characterization and genetic variations of a novel betanucleorhabdovirus infecting sugar beet in Iran. Pairwise comparison of the obtained betanucleorhabdovirus sequence with betanucleorhabdoviruses from GenBank showed that it shares the highest nucleotide identity with tomato betanucleorhabdovirus 2 (TBRV2) and Sambucus betanucleorhabdovirus 4. The obtained sequence contained six open reading frames (ORFs) in antigenomic sense flanked by complementary 3' leader and 5' trailer sequences (3'-N-P-P3-M-G-L-5'). In the phylogenetic tree, the detected isolate was clustered with the betanucleorhabdoviruses and was most closely related to TBRV2, tomato betanucleorhabdovirus 1, and Sambucus betanucleorhabdovirus 1 to 5. The genome organization, phylogenetic relationships, and sequence similarities to other betanucleorhabdoviruses suggest that the virus is a new member of the genus Betanucleorhabdovirus, which we propose to name beet betanucleorhabdovirus 1 (BNRV1). The virus's partial spread and host-range investigations revealed that sugar beet fields in northeast Iran were infected by BNRV1, which also infects Chenopodium album, Malva neglecta, and Carthamus oxyacanthus weeds. Low values of Ks*, Kst*, Z*, and Snn indicate no substantial genetic differentiation between populations of this virus. Selection pressures on a portion of the BNRV1 N gene analyzed were negative, showing purifying selection was occurring. The magnitude of negative selection in the BNRV1 N gene was consistent with what has been reported for other betanucleorhabdoviruses.
