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Making the best of a bad sample: Comparison of DNA extraction and quantification methods using sub-optimally stored Ixodes ricinus ticks

The primary aim of this study was to evaluate the performance of four DNA extraction methods on a collection of 200 sub-optimally stored Ixodes ricinus ticks from Southeastern Norway (177 nymphs, 11 males and 12 females). The methods were ammoniium hydroxide hydrolysis of homogenised or intact ticks and two commercial silica membrane-based kits, the QIAGEN Blood and Tissue kit and the QIAGEN Mini kit. DNA was evaluated by spectrophotometry, fluorometry, agarose gel electrophoresis and quantitative PCR. The second aim was to compare methods of evaluating the yield and purity. All four extraction methods provided amplifiable DNA, but the yield was variable (median 151 ng, range 0.13 to 4500 ng). DNA yields were not significantly different across the methods. Nine of 200 samples were inhibitory, all of which were ammonium hydroxide extracts of homogenised ticks. DNA purity, as judged by A260/280 ratios, was low; it was highest (mean = 1.63) for the Qiagen Blood and Tissue kit with the other methods showing values averaging 1.44. DNA yield measurements using qPCR, fluorometry (Qubit), drop spectrophotometry (NanoDrop) and gel electrophoresis (E-gels) correlated poorly, r ranging from <0 to 0.9 with no systematic pattern (average 0.4), probably reflecting the effects of low purity, low concentrations and differing amounts of single- and double-stranded DNA. Neoehrlichia mikurensis and Borrelia burgdorferi s.l. were detected in 5 and 13% of samples, respectively by qPCR. Our findings indicate that for the purposes of qPCR analysis, ammoniium hydroxide hydrolysis of intact ticks, a very cheap and simple method is as good as any of the other methods tested.

Polycross breeding enhances cumin quality and drought tolerance for sustainable agriculture

Constitutively active glucagon receptor drives high blood glucose in birds

The effect of traditional Thai massage vs routine physical therapy on gait pattern in spastic cerebral palsy: A cross-over randomized controlled trial

Background Various types of massage, including the traditional Thai, have not yet provided conclusive evidence to reduce the spasticity and improve walking ability for cerebral palsy (CP). Objectives To assess the effect of traditional Thai massage (TM) vs. a standard physical therapy (PT) on gait pattern in spastic CP. Methods A cross-over single blinded randomized controlled trial was conducted between October 2022 and October 2023 (Thai Clinical Trials Registry: TCTR20220530007). Individuals with aged 5 years or older, and diagnosed as spastic CP, Gross Motor Function Classification System (GMFCS) I-III were recruited. Participants were randomly assigned into group A (TM followed by PT), and group B (PT followed by TM). Each treatment lasted for 6 weeks (Phase I-II), with a 6-week washout period. Lower extremity range of motion, muscle tone, electromyography, gait profile score (GPS), oxygen consumption was blindly assessed and intention-to-treat analyzed. Results From 32 eligible patients (16 cases per group), average age was 16.1 ± 9.8 years (group A), and 13.6 ± 5.8 years (group B). Group B demonstrated higher left ankle dorsiflexion than group A (1.7 ± 13.0 degrees vs 12.1 ± 6.9 degrees, P-value = 0.011). GPS slightly improved in Phase I, and contradictory enhanced in TM while deteriorated in PT in Phase II without significant differences between groups. After adjusted for ankle dorsiflexion, TM significantly provided less overall GPS (-1.6 (95% confidence interval (CI) -2.6, -0.6), P-value = 0.001), and higher peak activity of right rectus femoris (0.132 mV (95%CI 0.001, 0.262), P-value = 0.030) compared to PT. Other outcomes did not differ significantly between two treatments, and no complication was detected. Conclusions TM has a positive effect on gait performance, as indicated by GPS and increased activity in the right rectus femoris muscle when compared to PT. A large-scale non-inferiority trial is required to affirm the impact of TM.

Application of RNA-seq for single nucleotide variation identification in a cohort of patients with hypertrophic cardiomyopathy

Abstract A variety of techniques for DNA sequencing, such as specific gene sequencing, whole genome sequencing, or exome sequencing, are currently used to detect single nucleotide variations (SNVs). Although RNA-seq can be used to identify SNVs, studies that employ this approach are uncommon, and those that do often rely on outdated mapping methods or methods that are more suitable for genomic and exomic alignment. In this work, our aim is to apply modern RNA-seq specific alignment method in order to identify SNV in a cohort of HCMP patients, and characterize those SNV to gain insight into possible mechanisms of HCMP pathogenesis. The algorithm of identification of SNV based on transcriptomic sequencing data has been developed and evaluated. The algorithm was evaluated and the optimal quality threshold was determined based on allelic discrimination for the rs397516037 mutation (MYBPC3 c.3697 C > T) among patients. A total of 42,809 SNVs with a quality of 75 or higher were identified in 48 transcriptomes of hypertrophic cardiomyopathy (HCMP) myocardial tissue. Verification of missense and nonsense variants in key HCMP genes using Sanger sequencing confirmed the accuracy of the pipeline results. To identify variants potentially associated with HCMP pathogenesis, a filtration process was conducted based on minor allele frequency, substitution prediction score and ClinVar outcome. 214 missense mutations and 6 nonsense mutations were selected. Together with nonsense mutations, 19 mutations meeting the strictest SIFT and PolypPhen criteria were identified as potential factors influencing HCMP pathogenesis. We have developed and validated a method for identifying SNVs based on transcriptomic data, which can be used to identify putative pathogenic variants. We identified mutations in key HCMP genes MYBPC3 and MYH7 in a cohort of patients. We also found potentially pathologic mutations in genes ANXA6 and FEM1 A and obtained data supporting the role of NEBL in myocardial diseases. This method would be useful in analyzing transcriptomic data available in the Gene Expression Omnibus, but should be used with caution as we have tested it on a specific disease.

Food label literacy among Malaysian school adolescents: A prevalence study

Making informed food choices is crucial for all ages, especially adolescents who are involved in selecting and buying food. Understanding nutrition labels helps guide choices and ensures nutritional needs are met. This study aims to assess the prevalence and level of food label literacy among school-going adolescents aged 13–16 in Malaysia. This study involved a cross-sectional online survey and was sampled by random sampling involving secondary school adolescents aged 13–16, totaling 1983 subjects across five regions in Malaysia: Central, Southern, Northern, and East Coast of Peninsular Malaysia, as well as East Malaysia (Sabah and Sarawak). The questionnaire includes sections on sociodemographic and anthropometric information, as well as three main food label literacy domains: comprehension and interpretation of food labels, skills in using food labels, and attitudes towards food labels. A total of 89.7% of adolescents read food labels with a moderate level of literacy across all three domains (comprehension and interpretation, skills in usage, and attitudes) classified using predefined score ranges based on previous studies. A significant relationship (p < 0.05) was found between age and body mass index-for-age z-score (BAZ) with the level of food label literacy, indicating higher literacy values in older adolescents. Mean scores of all domains A, B, and C across the five regions indicated significant differences at F(4,1978) = 25.8, p = 0.000, F(4,1978) = 17.43, p = 0.000, and F(4,1978) = 25.20, p = 0.000. In conclusion, the proportion of Malaysian adolescents reading food labels is high (89.7%). Nevertheless, the food label literacy level across all three domains is moderate. Hence, the promotion of the importance of nutrition labels in schools through organised programs such as interactive hands-on label reading activities and school-based nutrition campaigns are recommended. A crucial gap in health education can be addressed by fostering proper comprehension, skills, and attitudes toward food labels and facilitating informed dietary practices among adolescents.

A deep learning model for prediction of lysine crotonylation sites by fusing multi-features based on multi-head self-attention mechanism

Gender perspectives on zoonotic disease epidemiology; A strength weakness opportunities threats analysis in Bundibugyo district, Uganda

Background Gender mainstreaming in zoonotic disease prevention and control is paramount to produce sustainable solutions as well as keeping communities at the human-animal-environment interface safe and healthy. It is important to note that zoonoses register high mortality rates globally once they occur and they are highly transmissible. Hence this study aimed to explore the gender perspectives on zoonotic disease epidemiology using a (strength, weakness, opportunities and threats) SWOT analysis in Bundibugyo district, Uganda. Methods This study employed a descriptive participatory approach, utilizing data gathered sequentially from 12 Key informant interviews, 4 In-depth interviews and 4 Focus group discussions methods in Bundibugyo district. Interviews used interdisciplinary groups systematically using a SWOT analysis. The research methodology employed an interpretative phenomenological analysis (IPA) approach allowing participants to articulate their perspectives in their own words. Data analysis was done using both inductive and deductive thematically using NVIVO 12 pro, facilitating the development of codes, sub-themes, and themes. Results The results of this study prioritized key zoonotic diseases within the district that affect community health. The key themes from these results a) Threats increasing risk of zoonotic spillover, b) Community weaknesses increasing zoonotic diseases occurrence, c) Community strength for zoonotic disease management, prevention and control, d) Opportunities for communities in management of zoonotic diseases. The study emphasizes that political instability, land migration, food insecurity, cultural hunting practices, and climate change act to increase the risk of zoonotic diseases. Complications arise from the mis-identification of diseases due to similar symptoms, and a lack of community education about these diseases. The risk of exposure is influenced by gender roles, with men, who generally interact more with animals, at higher risk. Conversely, women, due to their roles in caring for the sick and involvement in child immunization, are at risk but also play a crucial role in disease control. Despite these challenges, there are opportunities for disease management and prevention such as leveraging experienced health workers for disease identification and education, utilizing local communication channels, engaging opinion leaders for effective risk communication, and providing regular training for health workers could address these issues. However, limited funding hinders the execution of recommended strategies such as regular surveillance, tracing of suspected cases, and health register reviews. Conclusion This study emphasizes the necessity of gender-sensitive approaches in understanding and mitigating zoonotic diseases, advocating for strategies that recognize socio-cultural factors, promote health education, and tailor interventions to provide comprehensive care and protection for all, irrespective of gender.

Optimization of robotic spray painting trajectories using machine learning for improved surface quality

Abstract The production process needs spray painting particularly within automobile manufacturing since product painting accuracy establishes product quality. The combination of hand spray techniques produces intricate designs as well as small quantity needs yet industrial robots excel at painting large industrial product orders. Taguchi Design of Experiments (DoE) is used to investigate the effect of six process variables which included spray distance along with pressure, temperature, humidity level, speed and viscosity rate. Experiments were conducted via industrial robotic spraying with subsequent statistical evaluation through ANOVA tests and regression calculations. The research shows that viscosity together with temperature stands as primary influential factors for thickness deviation, yet speed and temperature jointly determine surface roughness outcomes. The predictive model performed with substantial accuracy based on its ability to achieve R² values of 0.9224for surface roughness measurements and 0.9707 for thickness variation determination. The study offers clear guidelines for practitioners to enhance their processes to produce high-quality products and time efficiency.

The analysis of waste heat recovery in steel enterprises’ data centers based on the Co-ah cycle

To improve the energy economic efficiency of Data Centers (DCs) in steel enterprises, a centralized heating scheme for waste heat recovery based on the Co-ah cycle is proposed. This scheme establishes a thermoelectric connection between the self-owned power plant of the steel enterprise and the DC, creating a waste heat recovery centralized heating system for a 15 MW DC. The energy efficiency indicators, environmental benefits, economic feasibility, and adaptability of the system are evaluated. The results show that the system can effectively recover waste heat from the DC, significantly reducing cooling electricity consumption during the heating season and decreasing original heating steam consumption by about 25%. Compared to DC using free cooling, the annual operating cost is reduced by 9.7%, with a dynamic payback period for equipment of 6–7 years. The system saves 3,671.5 tons of standard coal and reduces CO2 emissions by 1,615 tons annually compared to DC using isolated free cooling and traditional heating systems. The Improved Power Usage Effectiveness (PUE’) of the system is 1.195, and the Energy Reuse Effectiveness (ERE) is 0.769, outperforming the free cooling’s index of 1.341, although Exergy Reuse Effectiveness (ExRE) is slightly higher than that of free cooling. This system offers mutual benefits for self-owned power plant, DC, and heating companies, achieving a win-win operational state through suitable energy trading prices. The research conclusion provides valuable reference for the future investment and operation of DCs in steel enterprises.

Initial cosmetic outcome six months after kilovoltage therapy of facial basal cell carcinoma: A Danish national prospective study of 932 patients

Heat shock transcription factor 2 reduces mitochondrial pathway apoptosis in intestinal epithelial cells by inhibiting the increase in mitochondrial membrane permeability in ulcerative colitis

The destruction of intestinal mucosal mechanical barrier homeostasis caused by excessive apoptosis of intestinal epithelial cells (IECs) is an important reason for the occurrence and development of ulcerative colitis (UC). The increase in mitochondrial membrane permeability caused by the opening of the mitochondrial membrane permeability transition pore (mPTP) is a key link in the initiation of mitochondrial pathway apoptosis. Our previous studies revealed that heat shock transcription factor 2 (HSF2), which is highly expressed in the intestinal mucosa of UC patients, can inhibit the expression of the cytochrome C (Cyto-C)/Caspase-9/Caspase-3 proteins in the mitochondrial pathway of apoptosis, but the regulatory mechanism is unknown. It has been reported that heat shock proteins regulated by heat shock transcription factors are closely related to mPTP opening. Therefore, we hypothesized that HSF2 affects mitochondrial pathway apoptosis in IECs by regulating mPTP opening. In this study, we altered the level of HSF2 in Caco-2 cells by lentivirus transfection to explore the changes in the mitochondrial membrane permeability of Caco-2 cells in an inflammatory environment. Subsequently, the mPTP agonist atractylorhizin (Atr) and inhibitor cyclosporine A (CsA) were used to clarify the regulatory effects of HSF2 on mPTP and the Cyto-C/Caspase-9/Caspase-3 pathways. Our study confirmed for the first time that HSF2 plays a protective role in UC by inhibiting mPTP opening, the increase in mitochondrial membrane permeability and the activation of the mitochondrial-mediated apoptosis pathway in IECs.

Development of new Eu3+/Sm3+ co-doped CaWO4 and CaMoO4 phosphors for optical thermometry based on ratiometric approach

Numerical study on the supersonic gas-solid two-phase injection mechanism of needle-free syringe

Supersonic gas-solid injection technology finds extensive use in drug particle delivery systems. However, the combined impact of particle diameter and mass flow rate on the delivery efficiency remain insufficiently explored. Within the Euler-Lagrange framework, this study utilizes the discrete phase method (DPM) for the numerical simulation of supersonic gas-particle flow in a needle-free injector. After validating the model’s accuracy with experiment results, further investigations were conducted into the influences of particle size and mass flow rate on particle behavior and flow field properties. The results indicate that the impact of larger particles on the compressible structure is stronger, while higher mass flow rate absorbs greater energy from the gas phase, reducing the gas expansion capacity, which results in lower velocity, Mach number, and higher temperature. The jet core zone is approximately x/X = 0.3 in length. Outside core zone, the gas velocity rapidly decays and temperature rises sharply. Within the jet core zone, drug particles are accelerated and cooled, while beyond core zone, they decelerate and heat up. The strongest inter-phase interactions occur primarily in the nozzle expansion area and the jet core zone. Smaller particles reach maximum velocity upstream. This implies that in designing needle-free injectors, the nozzle-to-skin distance must match the drug particle diameter to achieve maximum penetration effectiveness. Furthermore, the particle temperature decreases with smaller sizes. As the particle diameter rises from 10 μm to 100 μm, the minimum temperatures of the particles are 145 K and 264 K, respectively, indicating the need to match the particle diameter with the minimum temperature at which the drug particles remain effective. Additionally, higher mass flow rate doses reduce injection velocity and penetration ability, necessitating the rational control of the administered dose range. These results offer significant theoretical guidance for the design and improvement of needle-free injection.

Automated vacuum drying kinetics, thermodynamics, and economic analysis of sage leaves

Abstract Vacuum drying of sage leaves is important for preserving their essential oils, flavor, and medicinal properties by reducing oxidation and thermal degradation, but previous research has not investigated its impact on drying speed, thermodynamic properties, mathematical modeling, or economic viability. This study employed an automatic vacuum dryer at temperatures of 40 °C, 50 °C, and 60 °C under different pressure conditions (atmospheric, -5 kPa, and − 10 kPa) with a 1 cm layer thickness. Results showed that increasing temperature and decreasing pressure significantly improved drying efficiency, reducing the process time to just 90 min while achieving a drying rate of 22.34 kg water/kg dry matter/h and an effective moisture diffusivity of 6.716 × 10⁻⁹ m²/s under optimal conditions (60 °C and − 10 kPa). The Page model was identified as the most suitable for describing the thin-layer drying behavior. Thermodynamic analysis revealed activation energy values between 19.4 and 37.7 kJ/mol, with activation enthalpy decreasing at higher temperatures and lower pressures. The negative activation entropy values indicated chemical adsorption or structural modifications during drying. From an economic perspective, the most efficient drying conditions reduced the payback period to less than two months, demonstrating strong commercial potential. These findings highlight the industrial promise of vacuum drying for herb processing, with future research opportunities in process optimization, application to other herbs, and sustainability assessments to further enhance efficiency and economic benefits.

Deep-embedded clustering by relevant scales and genome-wide association study in autism

Autism spectrum disorder (ASD) presents with heterogeneous phenotypic and genetic characteristics. Despite investigation into the molecular mechanisms underlying ASD, its etiology remains elusive. In our previous investigation within the Simons Simplex Collection (SSC), we noted increased signals through a genome-wide association study (GWAS) by clustering patients with ASD and reducing the sample size. This study seeks to validate our previous study in a different population, the Simons Foundation Powering Autism Research for Knowledge (SPARK) population, while probing further into the genetic architecture of ASD. We examined data from 2,079 white male subjects and 875 unaffected SPARK siblings. Our methodology encompassed cluster analyses, followed by traditional GWAS and cluster-based GWAS (cGWAS). No significant associations were observed in the conventional GWAS when comparing all patients with all controls. However, in the cGWAS, by comparing patients clustered by phenotypes with controls, we identified 27 chromosomal loci meeting the criteria of p  < 5.0 × 10 ⁻  8 . Remarkably, several of these loci were situated within or in proximity to genes previously implicated as candidates for ASD. Nonetheless, our previous study of the SSC population did not fully replicate the SPARK population. The absence of reproducibility suggests the possibility of false positives within the cGWAS results due to potential technical factors. However, the emergence of multiple signals post-clustering and the association of numerous identified gene regions with ASD and related disorders provide supporting evidence for the validity of cGWAS outcomes.

Association of different treatment modalities for rectal cancer and second primary urinary system neoplasms, findings from SEER

Identification and validation of intergeneric hybrids between Saccharum officinarum and Erianthus rockii using molecular and cytogenetic tools

Sugarcane (Saccharum spp.), a vital crop for sugar and bioenergy production, faces challenges in breeding due to its narrow genetic base and susceptibility to environmental stresses. To enhance genetic diversity, distant hybridization with wild relatives such as Erianthus rockii, known for its drought tolerance and strong ratooning ability, offers a promising strategy. However, identifying true intergeneric hybrids remains a critical challenge. This study identified true intergeneric hybrids from crosses between S. officinarum and E. rockii using tetra-primer ARMS-PCR and HRM curve analysis targeting SNPs in the nrDNA-ITS region, with genomic in situ hybridization (GISH) validating chromosome composition. The results confirmed 13 true hybrids among 16 progeny, showing a chromosome inheritance pattern of 40 chromosomes derived from S. officinarum and 15 from E. rockii. Hybrid plants, while exhibiting traits from both parents, displayed inferior yield and sugar content compared to the maternal parent, indicating the necessity for further backcrossing to improve agronomic performance. This study pioneers the application of ARMS-PCR and HRM in identifying E. rockii hybrids and underscores their potential in advancing sugarcane breeding by facilitating the introgression of beneficial traits from wild relatives.

Long-term water immersion of dental composites based on bioactive glass

The motivational foundations of childbearing: Investigating relationships between childbearing attitudes, value preferences, and parental bonding

This study aims to reveal the motivational foundations of childbearing. To achieve this goal, we investigated how general motivational goals expressed in personal value preferences and relationships with one’s parents are connected to attitudes toward childbearing. The study is based on Schwartz’s theory of values [1]. The study was conducted in Israel among young adults (age 18–35) with no children (n = 1122). Higher preferences for tradition and benevolence-care and lower preferences for self-direction and universalism-nature values were associated with positive childbearing attitudes among men and women. In addition, higher preferences for security-social values were associated with positive childbearing attitudes among men. Maternal bonding was associated with positive childbearing attitudes among men and women, and paternal bonding was associated with positive childbearing attitudes among men. High religiosity, high income, and low education were associated with positive childbearing attitudes among women. Among men, only the effect of religiosity was significant. No difference between men and women in childbearing attitudes was found. Personal value preferences partly mediated the effects of parental bonding and socio-demographic variables on childbearing attitudes. The obtained results advance our understanding of the psychological mechanisms of childbearing.