Search Results (13,355)

Background: Tic spectrum disorder (TSD), encompassing Tourette syndrome and chronic tic disorder, is a childhood-onset neurodevelopmental condition with complex genetic and environmental contributions. Heritable components have been implicated in TSD, but no clear genetic mechanisms have been identified. Significant aspects of TSD etiology remain unclear, with key uncertainties concerning the role of environmental influences in its development. In this study, we aimed to identify environmentally induced epigenomic and transcriptomic changes contributing to TSD pathology by investigating genetically similar monozygotic twins discordant for TSD. Methods: To investigate environmentally driven mechanisms, we analyzed peripheral blood from eleven monozygotic twin pairs, either discordant or concordant for TSD, using RNA sequencing and DNA methylation analysis. Results: Differential expression analysis identified a dozen differentially expressed genes between TSD and non-TSD individuals, most of which were long non-coding RNAs or pseudogenes. Expression of the small RNA gene RNY1 was significantly associated with tic severity, suggesting involvement of immune-related processes. DNA methylation (DNAm) analysis revealed ~30,000 probes with a nominal p < 0.05, however none of these were significant after multiple testing correction. Expression quantitative trait methylation (eQTM) analysis identified 236 methylation-associated genes. Gene set enrichment analysis demonstrated broad downregulation in TSD individuals for pathways related to translation, RNA processing, and neurobiological functions, with Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways including ribosome, nucleocytoplasmic transport, pluripotency signaling, and nicotine addiction. Conclusions: These results suggest that environmentally influenced gene expression may contribute to TSD pathogenesis through dysregulation of immune and neuronal pathways. Despite a small sample size, the monozygotic twin design provides strong control for genetic background and identifies significant differences that contribute to the understanding of the underlying molecular mechanisms of TSD. Full article

In recent years, diagnostic technologies that utilize noninvasively collected sweat have garnered significant interest. However, the concentration of components in sweat is lower than that in blood, making the introduction of a concentration step as a sample pretreatment crucial for achieving highly sensitive detection. In this study, we developed a PDMS-based microchannel filled with silica gel, a desiccant particle, to concentrate liquid samples at room temperature without requiring an external power source or heating. The evaluation of the basic characteristics of the fabricated microchannel confirmed that filling it with silica gel efficiently removed the solvent vapor from the liquid samples. In concentration tests using the fluorescent dye uranine as a model for sweat sugar, a maximum 1.4-fold concentration was achieved in DPBS solution and a 1.2-fold concentration in artificial sweat at room temperature. In contrast, no similar concentration effect was observed in microchannels without silica gel packing. The proposed silica-gel-packed PDMS microchannel features a simple structure and requires no external equipment, making it easily integrable with existing microfluidic devices as a sample pretreatment module. This method is considered useful as a passive and simple sample concentration technique for the analysis of low-molecular-weight components in sweat. Full article

A new generation of insecticide-treated nets (ITNs) that incorporate the synergist piperonyl butoxide (PBO) has been shown to restore susceptibility to pyrethroids where P450 enzymes are the primary mechanism conferring the resistance. The present study evaluated the efficacy of YAHE 4.0, a PBO ITN, against wild free-flying Anopheles arabiensis in experimental huts in Lower Moshi, north-eastern Tanzania. It is the first evaluation of YAHE 4.0 in the country. Bio-efficacy evaluations, including susceptibility tests and cone bioassays, were conducted using the standard WHO guidelines. DuraNet Plus, a WHO-recommended PBO ITN, and Interceptor ITNs served as active and standard comparators, respectively. Unwashed and 20 times washed nets were subjected to experimental hut trials. Multiple logistic regression was employed to analyse experimental hut trial data. The results of the susceptibility testing showed that the An. arabiensis population of Lower Moshi was resistant to pyrethroids, but susceptible to organophosphates. Particularly, low mortality was recorded for cyhalothrin (2%) and alpha-cypermethrin (38%). Mortality rates to alpha-cypermethrin pirimiphos-methyl were 38% and 100%, respectively. The non-inferiority of YAHE 4.0 to DuraNet Plus ITN in terms of mortality and blood feeding was determined according to the WHO guidelines. The results for pooled unwashed and 20 times washed ITNs showed that YAHE 4.0 was superior to Interceptor ITN (adjusted odds ratio, AOR = 1.33; 95% CI = 1.04–1.69; non-inferiority margin, NIM = 0.68; p-value = 0.023) and non-inferior to DuraNet Plus (AOR = 1.02; 95% CI = 0.78–1.35; NIM = 0.72; p-value = 0.867) in terms of mortality. In terms of blood feeding inhibition for pooled unwashed and 20× washed ITNs, YAHE 4.0 was superior to both Interceptor ITN (AOR = 0.80; 95% CI = 0.64–1.00; NIM = 1.35; p-value = 0.049) and DuraNet Plus (AOR = 0.67; 95% CI = 0.52–0.86; NIM = 1.33; p-value = 0.002). Chemical analysis showed higher wash retention of active ingredients in YAHE 4.0 LLIN (88.9% for PBO and 94.9% for alpha-cypermethrin) compared to DuraNet Plus LLIN (89.2% for PBO and 90.5% for alphaypermethrin) before the hut trial. YAHE 4.0 LLIN demonstrated superior entomological efficacy and wash durability to DuraNet Plus and Interceptor LLINs, and fulfilled WHO non-inferiority criteria for mosquito mortality and blood-feeding inhibition. Therefore, YAHE 4.0 LLIN should be considered as an addition to the current list of pyrethroid-PBO nets used for control of pyrethroid-resistant vector populations with P450 enzymes as the main mechanism conferring resistance. Full article

Pregnancies complicated by diabetes, including pregestational and gestational diabetes mellitus, are associated with increased maternal and fetal morbidity. Early identification of at-risk pregnancies is crucial for timely intervention and improved outcomes. Emerging evidence highlights the interplay of genetic predisposition, epigenetic modifications, and non-invasive biomarkers in the early detection of diabetic pregnancies. Genetic factors influencing insulin signaling, glucose metabolism, and pancreatic β-cell function may contribute to susceptibility to gestational hyperglycemia. Concurrently, epigenetic alterations, such as DNA methylation and histone modifications in maternal and placental tissues, have been linked to dysregulated metabolic pathways and adverse pregnancy outcomes. Non-invasive biomarkers, including circulating cell-free DNA and microRNAs in maternal blood, show promise for early diagnosis by offering a safer and more practical alternative to invasive testing. Integrating genetic, epigenetic, and molecular marker data could enhance risk stratification and enable personalized monitoring and management strategies. This review synthesizes current knowledge on the molecular underpinnings of diabetic pregnancies, evaluates the potential of emerging biomarkers for early diagnosis, and discusses the challenges and future perspectives for translating these findings into clinical practice. Understanding these mechanisms may pave the way for precision medicine approaches, ultimately improving maternal and neonatal outcomes in pregnancies affected by diabetes. Full article

Objective: Alzheimer’s disease (AD) and other forms of dementia are a heterogeneous group of neurodegenerative diseases characterized by progressive cognitive decline. Differential diagnosis between AD and other dementias is crucial for choosing the optimal treatment strategy. Currently, cerebrospinal fluid (CSF) analysis remains the most accurate diagnostic method, but its invasiveness limits its use. In this regard, the search for reliable biomarkers in the blood is an urgent task. Methods: The study included 31 dementia patients (23 women and 8 men) diagnosed via interdisciplinary consultations and neuropsychological testing (MMSE ≤ 24). CSF and blood plasma samples were collected and analyzed using Luminex technology. Biomarker concentrations were measured, and statistical analyses (ANOVA, Kruskal–Wallis, and Pearson correlation) were performed to compare groups and assess correlations. Results: Levels of Aβ40 and Aβ42 in CSF were significantly lower in patients with AD compared with non-AD dementia (p = 0.02 and p < 0.001, respectively). The Aβ42/40 ratio in CSF was higher in patients with non-AD dementia (p = 0.048). The concentration of Aβ42 in blood plasma was increased in patients with AD (p = 0.001). Positive correlations were found between Aβ42 in CSF and TDP-43 in plasma in non-AD dementia (r = 0.97, p < 0.001), as well as between neurogranin and TDP-43 in plasma in AD (r = 0.845, p < 0.001). Conclusions: The study demonstrates the potential of blood biomarkers, in particular Aβ42, for the differential diagnosis of AD and other forms of dementia. The discovered correlations between CSF and plasma biomarkers deepen the understanding of neurodegenerative processes and contribute to the development of noninvasive diagnostic methods. Full article

Microscopic examination of Giemsa-stained thick blood smears remains the reference standard for malaria diagnosis, but it requires specialized personnel and is difficult to scale in resource-limited settings. We present a lightweight, smartphone-based system for automatic detection of Plasmodium parasites in thick smears captured with mobile phones attached to a conventional microscope. We built a clinically validated dataset of 400 slides from Loreto, Peru, consisting of 8625 images acquired with three smartphone models and 54,531 annotated instances of Plasmodium vivax and P. falciparum across eight morphologic classes. The workflow includes YOLOv11n-based visual-field segmentation, rescaling, tiling into 640 × 640 patches, data augmentation, and parasite detection. Four lightweight detectors were evaluated; YOLOv11n achieved the best trade-off, with an F1-score of 0.938 and an overall accuracy of 90.92% on the test subset. For diagnostic interpretability, performance was also assessed at the visual-field level by grouping detections into Vivax, Falciparum, Mixed, and Background. On a high-end smartphone (Samsung Galaxy S24 Ultra), the deployed YOLOv11n model achieved 110.9 ms latency per 640 × 640 inference (9.02 FPS). Full article

Laboratory maintenance of mosquitoes is important for studying vector biology and transmission of diseases, and for testing vector control tools. Standard operating procedures require feeding larvae with commercial fish meal. However, for many insectaries in sub-Saharan Africa, the commonly used feeds are imported and accompanied by procurement challenges. Changing the larval feed abruptly without allowing the larvae to adapt to new brands of feed also leads to a decrease in mosquito colonies in the laboratory. We investigated locally acquired beans, maize, and dried herrings as alternate feeds for mosquito larvae reared under laboratory conditions. Four replicates for each treatment were prepared, each containing 100 first instar larvae of Anopheles gambiae Tiassalé mosquitoes. The larvae were introduced into 500 mL of dechlorinated tap water and maintained under standard environmental insectary conditions. The larvae were provided with 40 mg of the designated powdered feed—beans, maize, and herring fish—in single and combined treatments. Tetra^® goldfish meal was included as a control. The larval mortality, developmental time, and number of pupae were recorded to evaluate the effectiveness of the feeds. Adult mosquitoes were weighed and measured to assess fitness, and females from each treatment were blood-fed and allowed to lay eggs to evaluate fertility. Larval survival differed significantly across diets (Kruskal–Wallis, p = 0.01), with maize-fed larvae showing the highest mortality (41.3%) and those with herring-based diets the lowest. Pupation and adult emergence were poorest in the maize and maize–bean groups, while the maize–herring combination achieved the highest emergence (92.6%, p = 0.03). Although overall differences were detected among the groups, conservative pairwise tests did not pinpoint specific group contrasts, but effect size estimates suggested biologically meaningful patterns. Generally, adult body weight and wing length did not differ by treatment except in maize-fed males (β = 0.371 mm, p = 0.022). Herring fish-based diets consistently supported larval survival, timely development, and robust fecundity, whereas maize-based diets were nutritionally inadequate. These findings highlight herring fish-based diets as a sustainable and cost-effective alternative to commercial feeds for maintaining Anopheles mosquito colonies, with potential to strengthen vector research capacity in resource-limited laboratories. Full article

In this work, kaolin/chitin (K/Ch) composite aerogels with different mass ratios were successfully fabricated via a freeze–drying approach. The influence of kaolin content on the microstructure, properties and hemostatic performance of the composite aerogels was systematically investigated. The results demonstrated that the incorporation of kaolin endowed the chitin-based aerogels with tunable porous structures, excellent water absorption capacity (up to 4282% for K_0.25/Ch₂), and enhanced water retention (73.7% for K₂/Ch₂ at 60 min). Moreover, the K/Ch composite aerogels exhibited good biodegradability, no cytotoxicity (cell viability > 91.9%), and no hemolysis (hemolysis rate < 1.5% at all test concentrations). In vitro hemostatic evaluations revealed that the composite aerogels exhibited rapid blood coagulation (blood clotting time of 16 s for K₂/Ch₂) with a blood coagulation index (BCI) as low as 0.5%, which was attributed to the synergistic effect of the physical adsorption of chitin and the coagulation cascade activation by kaolin. These findings indicated that the K/Ch composite aerogels could be used as novel natural hemostatic materials for potential effective and rapid hemostasis. Full article

Background/Objectives: Despite clonal expansion during a primary immune response, or after subsequent antigen encounters, the frequency of memory B cells (B_mem) specific for an antigen remains low, making their detection difficult. However, unlike serum antibodies, which have a short half-life in vivo and thus require continuous replenishment to maintain stable titers, circulating B_mem are long-lived; they preserve immunological preparedness through their ability to rapidly engage in recall responses and differentiate into antibody-secreting cells (ASCs) upon antigen encounter. To this end, development of assays suited for the reliable detection of rare antigen-specific B_mem is critical and can provide insights into an individual’s antigen exposure history and immune status beyond that offered by traditional serum antibody measurements alone. Methods: ImmunoSpot^® has emerged as a suitable technique for the detection of individual antigen-specific B cells through visualizing their antibody-derived secretory footprints. Here, we report the theoretical and practical foundations for detecting rare antigen-specific B_mem in human peripheral blood mononuclear cells (PBMC). Leveraging the unique availability of verifiably naïve vs. antigen-experienced human samples, we used SARS-CoV-2 Spike (S-) and Nucleocapsid (NCAP) antigens to interrogate the presence of B_mem with these respective specificities. Results: While 100% diagnostic accuracy was achieved for both antigens, detection of NCAP-specific B_mem required reducing the lower detection limit of the standard assay. Specifically, this was achieved by testing a total of 2 million PBMC across multiple replicate assay wells and assessing the cumulative number of secretory footprints detected. Conclusion: The protocols described here should facilitate the reliable detection of ASCs present at varying precursor frequencies and serve as guidance for routine immune monitoring of rare B_mem with specificity for any antigen. Full article

Hemotropic mycoplasmas (hemoplasmas) are uncultivable, cell wall-less bacteria that parasitizeon the surface of red blood cells of mammals, potentially causing anemia and other systemic signs. While widely distributed among domestic and wild animals, their occurrence in equids remains poorly understood, and no species has been identified as host-specific to horses or donkeys. This study presents the first systematic survey of hemoplasmas in equids from southeastern Europe and only the second molecularly confirmed case in horses in Europe. A total of 843 equids (817 horses and 26 donkeys) from different regions of Croatia, representing various ages, uses, and husbandry systems, were screened for hemoplasmas by PCR targeting the 16S rRNA gene. Only one horse tested positive, identified as Mycoplasma wenyonii, a hemoplasma typically associated with cattle. The estimated prevalence was 0.12% (95% CI: 0.003–0.68%). No donkeys were infected. The extremely low prevalence observed here—the lowest reported in any study detecting hemoplasma-positive horses—supports the hypothesis that equids do not harbor host-specific hemoplasma species and may only sporadically acquire infections from other hosts via spillover. This finding underscores the apparent absence of persistent hemoplasma lineages adapted to equids and highlights the need for further research on their epidemiology, host specificity, and transmission dynamics. Full article

Background: Irisin, a muscle-derived hormone, enhances the energy metabolism by activating the brown adipose tissue and acts as a bone-forming agent across the entire life span. No consistent clinical data in humans have been published so far to highlight if blood irisin as glucose/bone biomarker should be refined based on the vitamin D status (deficient or sufficient). Therefore, we aimed to objectively assess the level of irisin in female adults with abnormal and normal vitamin D status, as reflected by the level of 25-hydroxyvitamin (25OHD) in relationship with glucose and bone metabolic parameters. Methods: This pilot, prospective, exploratory study included eighty-nine menopausal women aged over 50. We excluded subjects with malignancies, bone and metabolic disorders, insulin treatment, and active endocrine disorders. Fasting profile included glycaemia, insulin, and glycated haemoglobin A1c (HbA1c). Then, 75 g oral glucose tolerance test (OGTT) included glycaemia and insulin assay after 60 and 120 min. Bone status involved bone turnover markers and central dual-energy X-ray absorptiometry providing bone mineral density (BMD) and trabecular bone score. Results: Eighty-nine subjects were included in the following two groups depending on 25OHD: vitamin D-deficient (VDD) group (N = 48; 25OHD < 30 ng/mL) and vitamin D-sufficient (VDS) group (N = 41; 25OHD ≥ 30 ng/mL). The two groups had similar age and menopausal period (62.29 ± 10.19 vs. 63.56 ± 8.16 years, respectively; 15.82 ± 9.55 vs. 16.11 ± 9.00 years, p > 0.5 for each). A statistically significant higher body mass index (BMI) was found in VDD vs. VDS group (32.25 ± 5.9 vs. 28.93 ± 4.97 kg/m², p = 0.006). Circulating irisin was similar between the groups as follows: median (IQR) of 91.85 (44.76–121.76) vs. 71.17 (38.76–97.43) ng/mL, p = 0.506. Fasting profile and OGTT assays showed no between-group difference. Median HOMA-IR in VDD group pointed out insulin resistance of 2.67 (1.31–3.29). Lowest mean/median T-scores at DXA for both groups were consistent with osteopenia category, but they were confirmed at different central sites as follows: femoral neck in both groups [VDD versus VDS group: −1.1 (−1.20–−0.90) vs. −1.1 (−1.49–−0.91), p = 0.526, respectively], only at lumbar spine for VDS group (T-score of −1.18 ± 1.13). The correlations between irisin and the mentioned parameters displayed a different profile when the analysis was performed in the groups with different 25OHD levels. In VDD group, irisin levels statistically significantly correlated with serum phosphorus (r = −0.32, p = 0.022), osteocalcin (r = −0.293, p = 0.038), P1NP (r = −0.297, p = 0.04), HbA1c (r = 0.342, p = 0.014), and BMI (r = 0.408, p = 0.003). Conclusions: This pilot study brings awareness in the analysis of irisin in relationship with glucose and bone-related biomarkers correlates, showing a distinct type of association depending on 25OHD level, which might represent an important crossroad in the multitude of irisin-activated signal transduction pathways. Full article

Pathogens such as Toxoplasma gondii, lentiviruses (e.g., CAE), Hypoderma spp., Neospora caninum, Mycoplasma spp., and pestiviruses are important for goat farming in Lithuania; however, data on their prevalence remain limited. To address this gap, a multi-pathogen study was conducted between 2021 and 2024 using selected ELISA kits (ID.vet, Innovative Diagnostics, France). A total of 380 blood samples were collected from 30 goat herds across different regions of Lithuania; the sample size varied depending on the pathogen. Serum samples were tested for antibodies, and seroprevalence was calculated for each pathogen. The highest seroprevalence was detected for T. gondii (38.9%, 143/368) and CAE virus (19.5%, 74/380). Antibodies to Mycoplasma spp. (0.3%, 1/368), Hypoderma spp. (3.8%, 7/184), and N. caninum (0.5%, 2/368) were detected only sporadically, while no antibodies to Border disease virus or Q fever were identified. Mixed infections were found in 7.6% of samples. Chi-square analysis showed that co-infections with toxoplasmosis and CAE occurred more frequently than expected (χ² = 19.05, p < 0.001). Herd size was significantly associated only with CAE seroprevalence (χ² = 7.913, df = 1, p < 0.05). Overall, toxoplasmosis and CAE were identified as the most epidemiologically relevant infections in the Lithuanian goat population. Full article

Vancomycin-resistant Enterococci (VRE) are established nosocomial pathogens; however, vancomycin-dependent Enterococci (VDE) represent a rare and underrecognized phenomenon. These organisms paradoxically require vancomycin for growth due to mutations in cell wall precursor synthesis. Limited awareness and significant diagnostic challenges associated with VDE can lead to delayed recognition and treatment failure. We report a case of vancomycin-dependent Enterococcus faecium isolated from a liver transplant recipient receiving oral vancomycin prophylaxis for recurrent Clostridioides difficile infection. The isolate failed to grow on standard media but exhibited robust growth on vancomycin-supplemented agar, confirmed by vancomycin disc diffusion testing and PCR detection of the vanB gene. Additionally, we reviewed four further VDE cases identified over a two-year period in our tertiary care microbiology laboratory. All patients originated from complex care settings, had significant comorbidities, and had received prolonged glycopeptide therapy. We summarize the clinical features, diagnostic findings, and microbiological challenges encountered across this case series. This series documents the first reported Canadian case of VDE and highlights the critical need for clinical vigilance and diagnostic suspicion in high-risk patients with prior enterococcal colonization and ongoing glycopeptide exposure. Laboratory findings such as failure to grow on blood agar coupled with growth around vancomycin discs should prompt specific evaluation for VDE. Our findings reinforce the necessity for targeted antimicrobial stewardship and infection prevention strategies and underscore the remarkable evolutionary adaptability of Enterococci under sustained antimicrobial pressure. Full article

Blood-sucking organisms produce various anticoagulant proteins that prevent blood clotting in their prey. Even in well-studied species like Hirudo medicinalis, many such proteins remain unidentified. We previously described a novel cysteine-rich anticoagulant (CRA), a distant homolog of antistasin. Later, we discovered another, much larger homolog in the medicinal leech. Its amino acid sequence is also highly cysteine-rich. Analysis of cysteine patterns showed four antistasin-like domain motifs, with one of them strongly disrupted. Since both antistasin and CRA contain two such domains, the new protein represents a duplicated antistasin-like structure. We cloned its cDNA, expressed the recombinant protein in Escherichia coli, purified it by metal-chelate chromatography, refolded it, and tested its anticoagulant properties. Using standard clinical assays—activated partial thromboplastin time, prothrombin time, and thrombin time—we found that the protein inhibited coagulation in all tests, though to varying degrees. These findings suggest that different antistasin-like anticoagulants in the leech enable it to block both intrinsic and extrinsic coagulation pathways, while hirudin inhibits the final step of clot formation. The combination of different anticoagulant proteins allows the leech to effectively prevent the prey’s blood from clotting during feeding. Full article

The forest fly (Hippobosca equina) is an obligate haematophagous dipteran insect (order Diptera) that primarily infests horses and may contribute to the circulation of vector-borne pathogens. This study aimed to investigate the presence of Anaplasma phagocytophilum, Borrelia burgdorferi s.l., Babesia caballi, and Theileria equi, important vector-borne pathogens of equids, in forest flies collected from horses in endemic areas of Spain. A total of 170 forest flies were collected from 39 equids across four geographical regions in Spain (Segovia, Madrid, Toledo, and Menorca) and blood samples were collected from 27 of these horses. All flies were morphologically and molecularly identified as H. equina, and DNA extracted from flies and equine blood was screened using multiplex real-time and nested PCR, followed by sequencing and phylogenetic analysis. Neither flies nor horses tested positive for A. phagocytophilum, whereas one fly was positive for B. burgdorferi s.l. (0.6%). In contrast, T. equi and B. caballi DNA were detected in 11.2% and 1.2% of flies, respectively, and all positive flies were collected from horses positive for equine piroplasmosis (T. equi/B. caballi infection), with identical 18S rRNA sequences between hosts and flies. Nested PCR showed a higher detection rate than real-time PCR for the detection of these piroplasms in flies and blood samples. These findings provide the first molecular evidence of EP pathogens in H. equina and support further investigation into the epidemiological importance of forest flies in equine pathogen surveillance. Full article