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Abstract Bloodstream infections (BSIs) and sepsis are major health problems, annually claiming millions of lives. Traditional blood culture techniques, employed to identify sepsis-causing pathogens and assess antibiotic susceptibility, usually take 2–4 days. Early and accurate antibiotic prescription is vital in sepsis to mitigate mortality and antibiotic resistance. This study aimed to reduce the wait time for sepsis diagnosis by employing shorter blood culture incubation times for BD BACTEC™ bottles using standard laboratory incubators, followed by real-time nanopore sequencing and data analysis. The method was tested on nine blood samples spiked with clinical isolates from the six most prevalent sepsis-causing pathogens. The results showed that pathogen identification was possible at as low as 102–104 CFU/mL, achieved after just 2 h of incubation and within 40 min of nanopore sequencing. Moreover, all the antimicrobial resistance genes were identified at 103–107 CFU/mL, achieved after incubation for 5 h and only 10 min to 3 h of sequencing. Therefore, the total turnaround time from sample collection to the information required for an informed decision on the right antibiotic treatment was between 7 and 9 h. These results hold significant promise for better clinical management of sepsis compared with current culture-based methods.

ABSTRACT Objective: Enlargement of the adrenal glands and variable adrenocortical function have been reported in patients with pulmonary tuberculosis and, in a few studies, in patients with extrapulmonary tuberculosis (EPTB). However, none of the studies have evaluated the course of the adrenal morphology in these patients. Subjects and methods: Prospective study including 37 patients with EPTB and 37 healthy age- and sex-matched controls. The adrenal function was evaluated by measurement of cortisol levels at baseline and after stimulation with ACTH (Acton Prolongatum) before and 6 months after antituberculosis treatment. The size of both adrenal glands was evaluated using 64-slice computed tomography (CT) scanning before and 6 months after treatment. The findings were compared with those in a group of healthy matched controls. Results: Clinical and biochemical parameters were comparable between groups. The mean baseline serum cortisol level was significantly lower in the EPTB group (397.1 ± 184.9 nmol/L) compared with the control group (696.3 ± 101.8 nmol/L). Compared with controls, patients with EPTB had significantly lower mean cortisol levels at baseline and 1 hour after ACTH, both before (397 ± 184.9 nmol/L and 750.7 ± 176.8 nmol/L, respectively) and after (529.7 ± 100.4 nmol/L and 1017.2 ± 119.7 nmol/L, respectively) antituberculosis treatment. Both the length and thickness of the right and left adrenal glands were greater in patients with EPTB than in controls but became comparable to those in controls after treatment completion. Conclusions: Patients with EPTB have an enlarged adrenal size and low baseline and stimulated serum cortisol levels. After treatment completion, cortisol levels increased significantly, and the adrenal size normalized in these patients.

SMK Negeri 1 Sidoarjo aims to create an automated hydroponic greenhouse system with IoT technology. Limited knowledge and costs hinder their progress. However, the Audio Video Engineering Department seeks assistance to achieve the program's goals by upgrading their knowledge and skills in automation. The Service Learning in Computer Engineering (SLiCE) program focuses on introducing and implementing automation and IoT technology in greenhouses. The community service project starts with surveys and discussions involving the principal and teachers. It then proceeds to analyze the needs and develop a hydroponic and IoT automation system. Training is provided for students and teachers of SMK Negeri 1 Sidoarjo. The ESP32 microcontroller enables remote monitoring and automatic control of nutrient levels and water pH, ensuring optimal plant growth. The outcome is a real-time hydroponic automation system capable of monitoring pH, nutrient PPM, and controlling nutrient addition through a website. After training, 87.9% of students reported improved comprehension of IoT and its implementation.

Background/Aim. The decreased mouth opening (MO) is a key factor in diagnosing most oral health-related issues. The aim of the study was to assess the magnitude of the maximum mouth opening (MMO) and its correlation with the facial types of children in the southern part of Saudi Arabia. Methods. A cross-sectional study was conducted in a group of 555 children ranging from 2 to 12 years of age. The MMO was measured as the distance between the incisal edge of maxillary and mandibular central incisors in the midline using the digital Vernier caliper. Three readings of MMO value were obtained, and the average mean of these readings was taken as the final MMO value. Based on the calculated facial index (FI), five facial types were determined in the study subjects: hypereuryprosopic (very broad face, FI range ≤ 79.9); euryprosopic (broad face, FI range 80– 84.9); mesoprosopic (round face, FI range 85–89.9); leptoprosopic (long face, FI range 90–94.9); hyperleptoprosopic (very l ong f ace, F I range ≥ 95). Subjects were categorized and compared accordingly. The facial profile, height, weight, and body mass index (BMI) of each subject were recorded as well. The data hence obtained was subjected to statistical analysis using SPSS (version 19.0, SPSS Incorporated, Chicago, IL, USA). Results. The overall mean value ± standard deviation of MMO for all the subjects in the study was 47.3 ± 8.7 mm. The mean value of MMO in hyperleptoprosopic children (FI range ≥ 95.0) was significantly higher than in other children. The mean difference value of MO in mesoprosopic (FI range 85.0–89.9) children was significantly lower than in leptoprosopic (FI range 90.0– 94.9) and hyperleptoprosopic (FI range ≥ 95.0) children. The mean difference value of MO of the leptoprosopic (FI range 90.0–94.9) type was significantly lower than that of the hyperleptoprosopic type (FI range ≥ 95.0). Conclusion. Based on the analyzed results, it was found that the MMO correlates with different facial types and facial profiles among the studied population.

Background: Rapid diagnostics of pathogens is essential to prescribe appropriate antibiotic therapy. The current methods for pathogen detection require the bacteria to grow in a culture medium, which is time-consuming. This increases the mortality rate and global burden of antimicrobial resistance. Culture-free detection methods are still under development and are not common in the clinical routine. Therefore, decreasing the culture time for accurately detecting infection and resistance is vital for diagnosis. Methods: This study investigated easy-to-implement factors (in a minimal laboratory set-up), including inoculum size, incubation temperature, and additional supplementation ( e.g., vitamin B12 and trace metals), that can significantly reduce the bacterial lag time (t lag). These factors were arranged in simple two-level factorial designs using Gram-positive cocci ( Staphylococcus aureus), Gram-positive bacilli ( Bacillus subtilis), and Gram-negative bacilli ( Escherichia coli and Pseudomonas aeruginosa) bacteria, including clinical isolates with known antimicrobial resistance profiles. Blood samples spiked with a clinical isolate of E. coli CCUG 17620 (Culture Collection University of Gothenburg) were also tested to see the effect of elevated incubation temperature on bacterial growth in blood cultures. Results: We observed that increased incubation temperature (42°C) along with vitamin B12 supplementation significantly reduced the t lag (10 – 115 minutes or 4% - 49%) in pure clinical isolates and blood samples spiked with E. coli CCUG17620. In the case of the blood sample, PCR results also detected bacterial DNA after only 3h of incubation and at three times the CFU/mL. Conclusion: Enrichment of bacterial culture media with growth supplements such as vitamin B12 and increased incubation temperature can be a cheap and rapid method for the early detection of pathogens. This proof-of-concept study is restricted to a few bacterial strains and growth conditions. In the future, the effect of other growth conditions and difficult-to-culture bacteria should be explored to shorten the lag phase.

Abstract Background The biological relevance and accuracy of gene expression data depend on the adequacy of data normalization. This is both due to its role in resolving and accounting for technical variation and errors, and its defining role in shaping the viewpoint of biological interpretations. Still, the choice of the normalization method is often not explicitly motivated although this choice may be particularly decisive for conclusions in studies involving pronounced cellular plasticity. In this study, we highlight the consequences of using three fundamentally different modes of normalization for interpreting RNA-seq data from human skeletal muscle undergoing exercise-training-induced growth. Briefly, 25 participants conducted 12 weeks of high-load resistance training. Muscle biopsy specimens were sampled from m. vastus lateralis before, after two weeks of training (week 2) and after the intervention (week 12), and were subsequently analyzed using RNA-seq. Transcript counts were modeled as (1) per-library-size, (2) per-total-RNA, and (3) per-sample-size (per-mg-tissue). Result Initially, the three modes of transcript modeling led to the identification of three unique sets of stable genes, which displayed differential expression profiles. Specifically, genes showing stable expression across samples in the per-library-size dataset displayed training-associated increases in per-total-RNA and per-sample-size datasets. These gene sets were then used for normalization of the entire dataset, providing transcript abundance estimates corresponding to each of the three biological viewpoints (i.e., per-library-size, per-total-RNA, and per-sample-size). The different normalization modes led to different conclusions, measured as training-associated changes in transcript expression. Briefly, for 27% and 20% of the transcripts, training was associated with changes in expression in per-total-RNA and per-sample-size scenarios, but not in the per-library-size scenario. At week 2, this led to opposite conclusions for 4% of the transcripts between per-library-size and per-sample-size datasets (↑ vs. ↓, respectively). Conclusion Scientists should be explicit with their choice of normalization strategies and should interpret the results of gene expression analyses with caution. This is particularly important for data sets involving a limited number of genes or involving growing or differentiating cellular models, where the risk of biased conclusions is pronounced.

Background: Rapid diagnostics of pathogens is essential to prescribe appropriate antibiotic therapy. The current methods for pathogen detection require the bacteria to grow in a culture medium, which is time-consuming. This increases the mortality rate and global burden of antimicrobial resistance. Culture-free detection methods are still under development and are not common in the clinical routine. Therefore, decreasing the culture time for accurately detecting infection and resistance is vital for diagnosis. Methods: This study investigated easy-to-implement factors (in a minimal laboratory set-up), including inoculum size, incubation temperature, and additional supplementation (e.g., vitamin B12 and trace metals), that can significantly reduce the bacterial lag time (tlag). These factors were arranged in simple two-level factorial designs using Gram-positive cocci (Staphylococcus aureus), Gram-positive bacilli (Bacillus subtilis), and Gram-negative bacilli (Escherichia coli and Pseudomonas aeruginosa) bacteria, including clinical isolates with known antimicrobial resistance profiles. Blood samples spiked with a clinical isolate of E. coli CCUG 17620 (Culture Collection University of Gothenburg) were also tested to see the effect of elevated incubation temperature on bacterial growth in blood cultures. Results: We observed that increased incubation temperature (42°C) along with vitamin B12 supplementation significantly reduced the tlag (10 – 115 minutes or 4% - 49%) in pure clinical isolates and blood samples spiked with E. coli CCUG17620. In the case of the blood sample, PCR results also detected bacterial DNA after only 3h of incubation and at three times the CFU/mL. Conclusion: Enrichment of bacterial culture media with growth supplements such as vitamin B12 and increased incubation temperature can be a cheap and rapid method for the early detection of pathogens. This proof-of-concept study is restricted to a few bacterial strains and growth conditions. In the future, the effect of other growth conditions and difficult-to-culture bacteria should be explored to shorten the lag phase.

Current state-of-the-art infection and antimicrobial resistance (AMR) diagnostics are based on culture-based methods with a detection time of 48–96 h. Therefore, it is essential to develop novel methods that can do real-time diagnoses. Here, we demonstrate that the complimentary use of label-free optical assay with whole-genome sequencing (WGS) can enable rapid diagnosis of infection and AMR. Our assay is based on microscopy methods exploiting label-free, highly sensitive quantitative phase microscopy (QPM) followed by deep convolutional neural networks-based classification. The workflow was benchmarked on 21 clinical isolates from four WHO priority pathogens that were antibiotic susceptibility tested, and their AMR profile was determined by WGS. The proposed optical assay was in good agreement with the WGS characterization. Accurate classification based on the gram staining (100% recall for gram-negative and 83.4% for gram-positive), species (98.6%), and resistant/susceptible type (96.4%), as well as at the individual strain level (100% sensitivity in predicting 19 out of the 21 strains, with an overall accuracy of 95.45%). The results from this initial proof-of-concept study demonstrate the potential of the QPM assay as a rapid and first-stage tool for species, strain-level classification, and the presence or absence of AMR, which WGS can follow up for confirmation. Overall, a combined workflow with QPM and WGS complemented with deep learning data analyses could, in the future, be transformative for detecting and identifying pathogens and characterization of the AMR profile and antibiotic susceptibility.

Background: The rapid diagnostics of pathogens is essential to prescribe appropriate and early antibiotic therapy. The current methods for pathogen detection require the bacteria to grow in a culture medium, which is time-consuming. This increases the mortality rate and the global burden of antimicrobial resistance. Culture-free detection methods are still under development and are not used in the clinical routine. Therefore decreasing the culture time for accurate detection of infection and resistance is vital for diagnosis. Methods: In this study, we wanted to investigate easy-to-implement factors (in a minimal laboratory set-up), including inoculum size, incubation temperature, and additional supplementation ( e.g., vitamin B12 and trace metals), that can significantly reduce the lag time (t lag). These factors were arranged in simple two-level factorial designs using Gram-positive ( Escherichia coli and Pseudomonas aeruginosa) and Gram-negative ( Staphylococcus aureus and Bacillus subtilis) bacteria, including clinical isolates with known antimicrobial resistance profiles. Blood samples spiked with a clinical isolate of E. coli CCUG17620 were also tested to see the effect of elevated incubation temperature on bacterial growth in blood cultures. Results: We observed that increased incubation temperature (42°C) along with vitamin B12 supplementation significantly reduced the t lag (10 – 115 minutes or 4% - 49%) in pure clinical isolates and blood samples spiked with E. coli CCUG17620. In the case of the blood sample, PCR results also detected bacterial DNA after only 3h of incubation and at three times the CFU/mL. Conclusions: Enrichment of bacterial culture media with growth supplements such as vitamin B12 and increased incubation temperature can be a cheap and rapid method for the early detection of pathogens. This is a proof-of-concept study restricted to a few bacterial strains and growth conditions. In the future, the effect of other growth conditions and difficult-to-culture bacteria should be explored to shorten the lag phase.

IntroductionRapid and accurate diagnosis of causative pathogens in mastitis would minimize the imprudent use of antibiotics and, therefore, reduce the spread of antimicrobial resistance. Whole genome sequencing offers a unique opportunity to study the microbial community and antimicrobial resistance (AMR) in mastitis. However, the complexity of milk samples and the presence of a high amount of host DNA in milk from infected udders often make this very challenging.MethodsHere, we tested 24 bovine milk samples (18 mastitis and six non-mastitis) using four different commercial kits (Qiagens’ DNeasy® PowerFood® Microbial, Norgens’ Milk Bacterial DNA Isolation, and Molzyms’ MolYsis™ Plus and Complete5) in combination with filtration, low-speed centrifugation, nuclease, and 10% bile extract of male bovine (Ox bile). Isolated DNA was quantified, checked for the presence/absence of host and pathogen using PCR and sequenced using MinION nanopore sequencing. Bioinformatics analysis was performed for taxonomic classification and antimicrobial resistance gene detection.ResultsThe results showed that kits designed explicitly for bacterial DNA isolation from food and dairy matrices could not deplete/minimize host DNA. Following using MolYsis™ Complete 5 + 10% Ox bile + micrococcal nuclease combination, on average, 17% and 66.5% of reads were classified as bovine and Staphylococcus aureus reads, respectively. This combination also effectively enriched other mastitis pathogens, including Escherichia coli and Streptococcus dysgalactiae. Furthermore, using this approach, we identified important AMR genes such as Tet (A), Tet (38), fosB-Saur, and blaZ. We showed that even 40 min of the MinION run was enough for bacterial identification and detecting the first AMR gene.ConclusionWe implemented an effective method (sensitivity of 100% and specificity of 92.3%) for host DNA removal and bacterial DNA enrichment (both gram-negative and positive) directly from bovine mastitis milk. To the best of our knowledge, this is the first culture- and amplification-independent study using nanopore-based metagenomic sequencing for real-time detection of the pathogen (within 5 hours) and the AMR profile (within 5–9 hours), in mastitis milk samples. These results provide a promising and potential future on-farm adaptable approach for better clinical management of mastitis.

Introduction: Dental caries is the most common chronic infectious disease involving children and toddlers. Carious invasion of anterior teeth causes infection, pain, compromised speech, mastication, and impacts the esthetics and appearance of the child. Dental caries' immediate and long-term effects affect the child's quality of life and his family, leading to significant social, economic, and psychological consequences. Aim: The present study was conducted to evaluate the effect of anterior dental caries on Saudi pediatric patients' psychological behavior and quality of life. Materials and Methods: A self-administered, structured questionnaire was used to assess the demographic variables, the prevalence of dental caries (anterior teeth), and their psychological effects on children. The data obtained were subjected to statistical analysis using IBM SPSS version 20.0 software. Results and Discussion: Five hundred and twenty-two children were included in the study, out of which 67.62% were affected with anterior carious lesions, with a mean age of 4.90 ± 1.981 years. Most parents reported that their child has anterior carious lesions, with 50% of children having toothache and 49% difficulty in eating food, especially sweets. Affected children showed psychological issues as they avoided playing, hesitated to smile, were introverted, and covered their faces while smiling. Conclusion: A high anterior dental caries prevalence and the resultant effect on psychological behavior and quality of life were observed among young children in Saudi Arabia. Enhanced educational, counseling, and preventive programs among parents and children will reduce dental caries incidence and ensure young children have a good quality of life.

Abstract Background Subjects with chronic obstructive pulmonary disease (COPD) are prone to accelerated decay of muscle strength and mass with advancing age. This is believed to be driven by disease-inherent systemic pathophysiologies, which are also assumed to drive muscle cells into a state of anabolic resistance, leading to impaired abilities to adapt to resistance exercise training. Currently, this phenomenon remains largely unstudied. In this study, we aimed to investigate the assumed negative effects of COPD for health- and muscle-related responsiveness to resistance training using a healthy control-based translational approach. Methods Subjects with COPD (n = 20, GOLD II-III, FEV1predicted 57 ± 11%, age 69 ± 5) and healthy controls (Healthy, n = 58, FEV1predicted 112 ± 16%, age 67 ± 4) conducted identical whole-body resistance training interventions for 13 weeks, consisting of two weekly supervised training sessions. Leg exercises were performed unilaterally, with one leg conducting high-load training (10RM) and the contralateral leg conducting low-load training (30RM). Measurements included muscle strength (nvariables = 7), endurance performance (nvariables = 6), muscle mass (nvariables = 3), muscle quality, muscle biology (m. vastus lateralis; muscle fiber characteristics, RNA content including transcriptome) and health variables (body composition, blood). For core outcome domains, weighted combined factors were calculated from the range of singular assessments. Results COPD displayed well-known pathophysiologies at baseline, including elevated levels of systemic low-grade inflammation ([c-reactive protein]serum), reduced muscle mass and functionality, and muscle biological aberrancies. Despite this, resistance training led to improved lower-limb muscle strength (15 ± 8%), muscle mass (7 ± 5%), muscle quality (8 ± 8%) and lower-limb/whole-body endurance performance (26 ± 12%/8 ± 9%) in COPD, resembling or exceeding responses in Healthy, measured in both relative and numeric change terms. Within the COPD cluster, lower FEV1predicted was associated with larger numeric and relative increases in muscle mass and superior relative improvements in maximal muscle strength. This was accompanied by similar changes in hallmarks of muscle biology such as rRNA-content↑, muscle fiber cross-sectional area↑, type IIX proportions↓, and changes in mRNA transcriptomics. Neither of the core outcome domains were differentially affected by resistance training load. Conclusions COPD showed hitherto largely unrecognized responsiveness to resistance training, rejecting the notion of disease-related impairments and rather advocating such training as a potent measure to relieve pathophysiologies. Trial registration: ClinicalTrials.gov ID: NCT02598830. Registered November 6th 2015, https://clinicaltrials.gov/ct2/show/NCT02598830

Abstract Background Lifestyle therapy with resistance training is a potent measure to counteract age‐related loss in muscle strength and mass. Unfortunately, many individuals fail to respond in the expected manner. This phenomenon is particularly common among older adults and those with chronic diseases (e.g. chronic obstructive pulmonary disease, COPD) and may involve endocrine variables such as vitamin D. At present, the effects of vitamin D supplementation on responses to resistance training remain largely unexplored. Methods Ninety‐five male and female participants (healthy, n = 71; COPD, n = 24; age 68 ± 5 years) were randomly assigned to receive either vitamin D3 or placebo supplementation for 28 weeks in a double‐blinded manner (latitude 61°N, September–May). Seventy‐eight participants completed the RCT, which was initiated by 12 weeks of supplementation‐only (two weeks with 10 000 IU/day, followed by 2000 IU/day), followed by 13 weeks of combined supplementation (2000 IU/day) and supervised whole‐body resistance training (twice weekly), interspersed with testing and measurements. Outcome measures included multiple assessments of muscle strength (nvariables = 7), endurance performance (n = 6), and muscle mass (n = 3, legs, primary), as well as muscle quality (legs), muscle biology (m. vastus lateralis; muscle fibre characteristics, transcriptome), and health‐related variables (e.g. visceral fat mass and blood lipid profile). For main outcome domains such as muscle strength and muscle mass, weighted combined factors were calculated from the range of singular assessments. Results Overall, 13 weeks of resistance training increased muscle strength (13% ± 8%), muscle mass (9% ± 8%), and endurance performance (one‐legged, 23% ± 15%; whole‐body, 8% ± 7%), assessed as weighted combined factors, and were associated with changes in health variables (e.g. visceral fat, −6% ± 21%; [LDL]serum, −4% ± 14%) and muscle tissue characteristics such as fibre type proportions (e.g. IIX, −3% points), myonuclei per fibre (30% ± 65%), total RNA/rRNA abundances (15%/6–19%), and transcriptome profiles (e.g. 312 differentially expressed genes). Vitamin D3 supplementation did not affect training‐associated changes for any of the main outcome domains, despite robust increases in [25(OH)D]serum (∆49% vs. placebo). No conditional effects were observed for COPD vs. healthy or pre‐RCT [25(OH)D]serum. In secondary analyses, vitamin D3 affected expression of gene sets involved in vascular functions in muscle tissue and strength gains in participants with high fat mass, which advocates further study. Conclusions Vitamin D3 supplementation did not affect muscular responses to resistance training in older adults with or without COPD.

Coronavirus disease 2019 (COVID-19), a mystified cryptic virus has challenged the mankind that has brought life to a standstill. Catastrophic loss of life, perplexed healthcare system and the downfall of global economy are some of the outcomes of this pandemic. Humans are raging a war with an unknown enemy. Infections, irrespective of age and gender, and more so in comorbidities are escalating at an alarming rate. Cardiovascular diseases, are the leading cause of death globally with an estimate of 31% of deaths worldwide out of which nearly 85% are due to heart attacks and stroke. Theoretically and practically, researchers have observed that persons with pre-existing cardiovascular conditions are comparatively more vulnerable to the COVID-19 infection. Moreover, they have studied the data between less severe and more severe cases, survivors and non survivors, intensive care unit (ICU) patients and non ICU patients, to analyse the relationship and the influence of COVID-19 on cardiovascular health of an individual, further the risk of susceptibility to submit to the virus. This review aims to provide a comprehensive particular on the possible effects, either direct or indirect, of COVID-19 on the cardiovascular heath of an individual.

The objective of this article was to systematically provide an up-to-date review on the different methods of remineralizing human dentine using different biomimetic agents. The authors performed a systematic search within PubMed, Scopus, and Web of Science in addition to the grey literature in Google Scholar® using MeSH terms. The PICO question was P: human teeth dentinal sections; I: application of biomimetic remineralizing agents; C: other non-biomimetic approaches; O: extent of remineralization and physical properties of remineralized dentine. The initially identified studies were screened for titles and abstracts. Non-English articles, reviews, animal studies, studies involving the resin–dentine interface, and other irrelevant articles were then excluded. The other remaining full-text articles were retrieved. Bibliographies of the remaining articles were searched for relevant studies that could be included. A total of 4741 articles were found, and finally, 39 full-text articles were incorporated in the current systematic review. From these, twenty-six research studies used non-collagenous protein (NCP) analogs to biomineralize dentine, six studies used bioactive materials derived from natural sources, six studies used zinc hydroxyapatite, and one study used amelogenin peptide to induce hydroxyapatite formation on the surface of demineralized dentine. Additive effects of triclosan and epigenin were assessed when combined with commonly available NCPs. Overall, a moderate risk of bias was observed and, hence, the findings of the included studies could be acceptable. A meta-analysis of some similar studies was performed to assess the depth of remineralization and elastic modulus. Despite having high heterogeneity (I2 > 90), all the studies showed a significant improvement in biomimetic remineralization efficacy as compared to the control. All the included studies carried out a functional remineralization assessment and found a 90–98% efficacy in the extent of remineralization while the elastic modulus reached 88.78 ± 8.35 GPa, which is close to natural dentine. It is pertinent to note the limitations of these studies that have been carried out in vitro under controlled settings, which lack the effects of a natural oral environment. To conclude, the authors suggest that the biomimetic remineralization of dentine using NCP analogs, bioactive materials, and natural products carries significant potential in treating dentinal lesions; however, more long-term studies are needed to assess their clinical applications in vivo.

Antimicrobial resistance (AMR), defined as the ability of microorganisms to withstand antimicrobial treatment, is responsible for millions of deaths annually. The rapid spread of AMR across continents warrants systematic changes in healthcare routines and protocols. One of the fundamental issues with AMR spread is the lack of rapid diagnostic tools for pathogen identification and AMR detection. Resistance profile identification often depends on pathogen culturing and thus may last up to several days. This contributes to the misuse of antibiotics for viral infection, the use of inappropriate antibiotics, the overuse of broad-spectrum antibiotics, or delayed infection treatment. Current DNA sequencing technologies offer the potential to develop rapid infection and AMR diagnostic tools that can provide information in a few hours rather than days. However, these techniques commonly require advanced bioinformatics knowledge and, at present, are not suited for routine lab use. In this review, we give an overview of the AMR burden on healthcare, describe current pathogen identification and AMR screening methods, and provide perspectives on how DNA sequencing may be used for rapid diagnostics. Additionally, we discuss the common steps used for DNA data analysis, currently available pipelines, and tools for analysis. Direct, culture-independent sequencing has the potential to complement current culture-based methods in routine clinical settings. However, there is a need for a minimum set of standards in terms of evaluating the results generated. Additionally, we discuss the use of machine learning algorithms regarding pathogen phenotype detection (resistance/susceptibility to an antibiotic).

Diseases of the oral cavity and musculoskeletal disorders (MSDs) are common occurrences. They are commonly linked with partial mobility, resulting in limited visits to dentists for routine oral care, causing poor periodontal condition, bleeding, gingival inflammation, and increased depth of periodontal pockets. The present study was conducted to measure joint movements, hand grip strength, and pain in joints and their association with oral health. Materials and methods: The study included 200 subjects, half suffering from back, neck, shoulder, elbow, and wrist problems, belonging to different age groups and professions; 100 had joint pain, and 100 were without joint pain. The study assessed individuals exposed to oral health issues by measuring the strength of hand grip, flexibility, and pain score of the back, neck, shoulder, elbow, and wrist. The strength of the hand grip and the angle of the elbow and shoulder were measured in addition to a questionnaire to assess the relationship between back pain and oral health. To evaluate dental health status and oral hygiene, the total number of decayed, missing, and filled teeth (DMF/T) and Oral Hygiene Index—Simplified (OHI-S) indices were used. Results: The difference between all demographic parameters was statistically significant (p < 0.05). It was observed that there was a significant difference in calculus, debris, and dental caries scores in both groups, with significantly lower scores and better oral hygiene in patients without joint pain. Concerning pain score and joint movements, the group without joint pain showed a significantly better range of movements and less pain than patients suffering from joint pain, and statistically a significant difference (p < 0.05) was observed between both groups. Conclusion: The present study revealed that musculoskeletal disorders, pain in the neck and hand, and restricted movements were common among professionals. We observed that pain in joints, neck, and hands, with restricted movements, caused a serious impact on the maintenance of oral hygiene practices among subjects of different professions.

Rapid bacterial identification and antimicrobial resistance gene (ARG) detection are crucial for fast optimization of antibiotic treatment, especially for septic patients where each hour of delayed antibiotic prescription might have lethal consequences. This work investigates whether the Oxford Nanopore Technology’s (ONT) Flongle sequencing platform is suitable for real-time sequencing directly from blood cultures to identify bacteria and detect resistance-encoding genes. For the analysis, we used pure bacterial cultures of four clinical isolates of Escherichia coli and Klebsiella pneumoniae and two blood samples spiked with either E. coli or K. pneumoniae that had been cultured overnight. We sequenced both the whole genome and plasmids isolated from these bacteria using two different sequencing kits. Generally, Flongle data allow rapid bacterial ID and resistome detection based on the first 1,000–3,000 generated sequences (10 min to 3 h from the sequencing start), albeit ARG variant identification did not always correspond to ONT MinION and Illumina sequencing-based data. Flongle data are sufficient for 99.9% genome coverage within at most 20,000 (clinical isolates) or 50,000 (positive blood cultures) sequences generated. The SQK-LSK110 Ligation kit resulted in higher genome coverage and more accurate bacterial identification than the SQK-RBK004 Rapid Barcode kit.

The aim of this study is to develop and to test a low-cost virtual reality spatial trainer in terms of its effectiveness in spatial training. The researchers adopted three features deriving from the constructivist perspective to guide the design of the trainer, namely interaction, instruction, and support. The no control pre test post test experimental procedure was used as the research design involving a sample, which comprised a class of 35 middle graders (15 girls and 20 boys) with mean age of 15.5 years. The researchers hypothesized that there would be a significant improvement in spatial visualization after training and the three training attributes would receive different ratings from the participants. The research instruments used were Spatial Visualization Test (for pre-testing and post-testing) and usability survey questionnaire. The participants used the trainer for four consecutive weeks, with each session lasting for three hours. Data collected were analyzed using Statistical Package for Social Science (SPSS). Result of a paired samples t-test indicated that the participants had made significant improvement after training, thus supporting the first research hypothesis. However, the results of independent samples t-test did not support the second hypothesis, where all the three training features were rated equally. Interestingly, subtle but significant orientations were detected, where the instructive and interactive features were rated differently by girls and boys, respectively. The researchers also discuss the practical implications of the low-cost virtual reality application for spatial training by proposing a set of design guidelines.

Antibiotic resistance poses a major threat to public health. More effective ways of the antibiotic prescription are needed to delay the spread of antibiotic resistance. Employment of sequencing technologies coupled with the use of trained neural network algorithms for genotype-to-phenotype prediction will reduce the time needed for antibiotic susceptibility profile identification from days to hours.In this work, we have sequenced and phenotypically characterized 171 clinical isolates of Escherichia coli and Klebsiella pneumoniae from Norway and India. Based on the data, we have created neural networks to predict susceptibility for ampicillin, 3rd generation cephalosporins and carbapenems. All networks were trained on unassembled data, enabling prediction within minutes after the sequencing information becomes available. Moreover, they can be used both on Illumina and MinION generated data and do not require high genome coverage for phenotype prediction. We cross-checked our networks with previously published algorithms for genotype-to-phenotype prediction and their corresponding datasets. Besides, we also created an ensemble of networks trained on different datasets, which improved the cross-dataset prediction compared to a single network.Additionally, we have used data from direct sequencing of spiked blood cultures and found that AMR-Diag networks, coupled with MinION sequencing, can predict bacterial species, resistome, and phenotype as fast as 1–8 h from the sequencing start. To our knowledge, this is the first study for genotype-to-phenotype prediction: (1) employing a neural network method; (2) using data from more than one sequencing platform; and (3) utilizing sequence data from spiked blood cultures.

Abstract Background Human skeletal muscle responds to weight-bearing exercise with significant inter-individual differences. Investigation of transcriptome responses could improve our understanding of this variation. However, this requires bioinformatic pipelines to be established and evaluated in study-specific contexts. Skeletal muscle subjected to mechanical stress, such as through resistance training (RT), accumulates RNA due to increased ribosomal biogenesis. When a fixed amount of total-RNA is used for RNA-seq library preparations, mRNA counts are thus assessed in different amounts of tissue, potentially invalidating subsequent conclusions. The purpose of this study was to establish a bioinformatic pipeline specific for analysis of RNA-seq data from skeletal muscles, to explore the effects of different normalization strategies and to identify genes responding to RT in a volume-dependent manner (moderate vs. low volume). To this end, we analyzed RNA-seq data derived from a twelve-week RT intervention, wherein 25 participants performed both low- and moderate-volume leg RT, allocated to the two legs in a randomized manner. Bilateral muscle biopsies were sampled from m. vastus lateralis before and after the intervention, as well as before and after the fifth training session (Week 2). Result Bioinformatic tools were selected based on read quality, observed gene counts, methodological variation between paired observations, and correlations between mRNA abundance and protein expression of myosin heavy chain family proteins. Different normalization strategies were compared to account for global changes in RNA to tissue ratio. After accounting for the amounts of muscle tissue used in library preparation, global mRNA expression increased by 43–53%. At Week 2, this was accompanied by dose-dependent increases for 21 genes in rested-state muscle, most of which were related to the extracellular matrix. In contrast, at Week 12, no readily explainable dose-dependencies were observed. Instead, traditional normalization and non-normalized models resulted in counterintuitive reverse dose-dependency for many genes. Overall, training led to robust transcriptome changes, with the number of differentially expressed genes ranging from 603 to 5110, varying with time point and normalization strategy. Conclusion Optimized selection of bioinformatic tools increases the biological relevance of transcriptome analyses from resistance-trained skeletal muscle. Moreover, normalization procedures need to account for global changes in rRNA and mRNA abundance.

Objective: The objective of the study is to understand the attitude and perceptions of children toward dentists, dental treatment, and clinical environment. Materials and Methods: A cross-sectional study was conducted where pictorial questionnaires were distributed among 498 children, within the age range of 3–14 years, who visited dental clinics in Aseer region of Saudi Arabia. Fisher's exact and Chi-square tests were applied to the collected data. Results: A majority (59.6%) of the children did like their visit to the dentists. About 63.1% of children preferred to be treated by a female dentist, and 53% of them liked their dentists wearing a colored coat. Female children were found to have significantly more positive responses toward protective gear of dentist and dental experience as compared to the males. Further, 9–11 years of children had more positive responses than the other children of different age groups. Conclusions: It was observed that most of the children liked visiting the clinics, and their siblings also shared their positive experience during dental treatment. However, children are found to fear the dental treatment procedure, especially the use of syringes. Fear toward dentists and clinical environment can be curbed using engaging dental practices. Children are found to be strongly affected by the gender and appearance of the dentists.

Abstract Background Sperm DNA integrity is considered essential for successful transmission of the paternal genome, fertilization and normal embryo development. DNA fragmentation index (DFI, %) has become a key parameter in the swine artificial insemination industry to assess sperm DNA integrity. Recently, in some elite Norwegian Landrace boars (boars with excellent field fertility records), a higher level of sperm DFI has been observed. In order to obtain a better understanding of this, and to study the complexity of sperm DNA integrity, liquid preserved semen samples from elite boars with contrasting DFI levels were examined for protamine deficiency, thiol profile and disulphide bonds. Additionally, the DNA methylation profiles of the samples were determined by reduced representation bisulphite sequencing (RRBS). Results In this study, different traits related to sperm DNA integrity were investigated (n = 18 ejaculates). Upon liquid storage, the levels of total thiols and disulphide bonds decreased significantly, while the DFI and protamine deficiency level increased significantly. The RRBS results revealed similar global patterns of low methylation from semen samples with different levels of DFI (low, medium and high). Differential methylation analyses indicated that the number of differentially methylated cytosines (DMCs) increased in the low-high compared to the low-medium and the medium-high DFI groups. Annotating the DMCs with gene and CpG features revealed clear differences between DFI groups. In addition, the number of annotated transcription starting sites (TSS) and associated pathways in the low-high comparison was greater than the other two groups. Pathway analysis showed that genes (based on the closest TSS to DMCs) corresponding to low-high DFI comparison were associated with important processes such as membrane function, metabolic cascade and antioxidant defence system. Conclusion To our knowledge, this is the first study evaluating DNA methylation in boar sperm cells with different levels of DFI. The present study shows that sperm cells with varying levels of DNA fragmentation exhibit similar global methylation, but different site-specific DNA methylation signatures. Moreover, with increasing DNA fragmentation in spermatozoa, there is an increase in the number of potentially affected downstream genes and their respective regulatory pathways.

This paper presents the systematic development of a "Uses and Gratification Expectancy" (UGE) conceptual framework which is able to predict students' "Perceived e-Learning Experience." It is argued that students' UGE as regards e-learning resources cannot be implicitly or explicitly explored without first examining underlying communication theories and learning perspectives. As such, the theoretical framework is grounded in the confluence of theories from communication theories and learning perspectives. The integration of Expectancy-value Theory, and the Uses and Gratification Theory serves to accommodate the suggestion that e-learning resources offer gratifications that are expected and valued by students. The key theoretical and practical assumptions of the UGE approach are highlighted and consistently implemented in the conceptual edifice. (Contains 6 figures.)

Taking advantage of the convergence of technology and new insights of teaching and learning, the Ministry of Education of Malaysia has implemented a new educational reform primarily to revise the current curriculum, establish new learning standards, and incorporate the use of technology as an integral part of the learning process. This new educational reform suggests more of a focus on critical-thinking processes, problem solving, and student assessment deemed critical in academic curricula. The purpose of this research was to examine factors that may influence students' perceived ability or self-efficacy to learn Engineering Drawing at the foundation year, namely in the Form Four level of the Malaysian secondary schools. The school system in Malaysia consists of primary and secondary levels. The Engineering Drawing course covers a two-year period, with fundamental concepts, theories, and techniques covered during the first year and advanced topics covered during the second year (11th grade). Several factors have been identified to be critical in shaping students' perceived ability to learn the subject matter. Three hypotheses were formulated from prior research: (1) High spatial experience students will have higher perceived ability than low spatial experience students to learn Engineering Drawing; (2) High mathematics achievers will have higher perceived ability than low mathematics achievers to learn Engineering Drawing; and (3) Male students will have higher perceived ability than female students to learn Engineering Drawing. In addition to the above hypotheses, the study also sought to identify the factor(s) among the three variables that may have a strong predictive power to explain the variation in the perceived ability of students when learning the subject. A total of two hundred and twenty four (224) students, including 75 females and 149 males of Form Four level (10th graders) taking the Engineering Drawing course at the foundation year participated in the study. This study employed a quantitative survey approach using two questionnaires. Findings from the study have provided several insights concerning the perceived ability of students to learn the subject matter of engineering drawing. Spatial experience was found to be a significant factor, having a substantial relationship with students' perceived ability to learn Engineering Drawing for all four categories of topics, thus confirming the first hypothesis of the study. Students with more spatial experience perceive that they will be able to learn the content of the course. Prior mathematics achievement was found to have a weak relationship in predicting students' perceptions on their ability to learn Engineering Drawing content overall. Thus the second hypothesis was only weakly supported. The notion that males would have a higher perception about their success in Engineering Drawing, the third hypothesis, was not supported. Females were found to have a higher perception than males about learning the topics, especially the low spatial content group.

Resin-based pit fissure sealants (RBS) are used to prevent occlusal caries in children. The success of RBS in preventing dental caries is largely influenced by its retention on the tooth surface, which is also affected by its physicomechanical properties. The physicomechanical properties of RBS can be enhanced through the addition of fillers. With the advent of nanofillers, the physicomechanical properties were improved without altering RBS flowability. The present study developed an RBS with a 70 wt% resin matrix and 30 wt% nanofillers. The resin matrix consisted of urethane dimethacrylate (55 wt%), triethylene glycol dimethacrylate (45 wt%), camphoroquinone (0.3 wt%), and 2-(dimethylamino) ethyl methacrylate (0.7 wt%). Silane-treated rice husk-derived nanosilica (20 wt%) and nanohydroxyapatite (10 wt%) were added as fillers. Clinpro, Fissurit FX, and an unfilled sealant were controls. All RBS were tested for surface roughness, Vickers hardness, flexural strength, and flowability. Statistical analysis with oneway ANOVA revealed significant differences between groups in surface roughness, hardness, flowability (p < 0.001), flexural strength, and flexural modulus (p < 0.05). Experimental sealants had higher flexural strength (78 MPa) and flow distance (29.05+1.16 mm) than commercial controls. However, the surface roughness of experimental sealants (0.25+0.08 µm) was higher than Clinpro (0.087+0.027 µm) but lesser than Fissurit FX (0.35+0.19 µm). The Vickers hardness of experimental sealants (23+1.63 VHN) was less than Fissurit FX (28.80+1.69 VHN) but higher than Clinpro (21.74+1.68 VHN). This novel RBS had physicomechanical properties comparable to commercial sealants. The use of nanosilica from rice husk makes this pit and fissure sealer sustainable and environmentally friendly in dentistry. [ABSTRACT FROM AUTHOR]

With increase in the understanding of stem-cell behavior, their application has extended well beyond not only regenerative medicine but also dentistry. The breakthrough of stem cells has produced new possibilities in the regeneration of different organs and tissues. In past few years many studies and revelations have been conducted in dentistry which indicate that stem cells and tissue engineering together are a unit have given rise to so called “Regenerative Dentistry”. The tooth (permanent teeth and deciduous teeth) is the natural store house for these stem cells. These are capable of reproducing themselves and can be voluntarily recovered at the time of an intended dental procedure. Besides dental stem cells have lot of medical benefits in the progress of new medical therapies. Like regenerative diseases and cancer therapy. This article highlights the research in dental field on Stem cells their current applications and what can be done and achieved in the future.