Thread-powered cell lysis and isotachophoresis: unlocking microbial DNA for diverse molecular applications.

Central to the domain of molecular biology resides the foundational process of DNA extraction and purification, a cornerstone underpinning a myriad of pivotal applications. In this research, we introduce a DNA extraction and purification technique leveraging polypropylene (PP) threads. The process commences with robust cell lysis achieved through the vigorous agitation of interwoven PP threads. The friction between the threads facilitates cell lysis especially those microbes having tough cell wall. For purification of DNA, thread-based isotachophoresis was employed which makes the whole process swift and cost-effective. Lysed cell-laden threads were submerged in a trailing electrolyte which separated DNA from other cellular contents. The process was performed with a tailored ITP device. An electric field directs DNA, cell debris, trailing electrolyte, and leading electrolyte toward the anode. Distinct ion migration resulted in DNA concentrating on the PP thread's anode-proximal region. The SYBR green dye is used to visualize DNA as a prominent green zone under blue light. The purified DNA exhibits high purity levels of 1.82 ± 0.1 (A260/A280), making it suitable for various applications aiming at nucleic acid detection.

Moving the Needle Forward in Genomically-Guided Precision Radiation Treatment.

Radiation treatment (RT) is a mainstay treatment for many types of cancer. Recommendations for RT and the radiation plan are individualized to each patient, taking into consideration the patient's tumor pathology, staging, anatomy, and other clinical characteristics. Information on germline mutations and somatic tumor mutations is at present rarely used to guide specific clinical decisions in RT. Many genes, such as ATM, and BRCA1/2, have been identified in the laboratory to confer radiation sensitivity. However, our understanding of the clinical significance of mutations in these genes remains limited and, as individual mutations in such genes can be rare, their impact on tumor response and toxicity remains unclear. Current guidelines, including those from the National Comprehensive Cancer Network (NCCN), provide limited guidance on how genetic results should be integrated into RT recommendations. With an increasing understanding of the molecular underpinning of radiation response, genomically-guided RT can inform decisions surrounding RT dose, volume, concurrent therapies, and even omission to further improve oncologic outcomes and reduce risks of toxicities. Here, we review existing evidence from laboratory, pre-clinical, and clinical studies with regard to how genetic alterations may affect radiosensitivity. We also summarize recent data from clinical trials and explore potential future directions to utilize genetic data to support clinical decision-making in developing a pathway toward personalized RT.

Carbon dots and their interactions with recognition molecules for enhanced nucleic acid detection.

Carbon Dots (C-dots) have exceptional fluorescence and incident wavelength alteration capabilities because of their π-π* electron transitions between the surface-trapped charges. They have clear, considerate and cost-effective applications in the domain of bio-sensing, optical imaging, medical diagnostics, fluorescence chemotherapy, forensics, and environmentology. Advances in the production process of C-dots can change their optical and chemical characteristics, allowing them to interact with a variety of chemicals and ions that can be exploited for the DNA detection in point-of-care devices. In the current scenario of pathogenic disease prevention, metagenomics and industrial processes, alternative genetic material identification is critical. This review focuses on the existing carbon dots-based DNA detection technologies and their interactions with other components such as metallic salts, dyes, and biological chemicals based on their surface charge distribution (positive or negative) employed in the DNA diagnostic devices and biosensors with their operating mechanism regarding their target component. These intriguing scientific discoveries and technologies will be extensively examined to translate them into real-world solutions which will have a significant societal and economic impact on overall well-being and innovation.

Quantification of Patient-Reported Pain Locations: Development of an Automated Measurement Method.

Patient-reported pain locations are critical for comprehensive pain assessment. Our study aim was to introduce an automated process for measuring the location and distribution of pain collected during a routine outpatient clinic visit. In a cross-sectional study, 116 adults with sickle cell disease-associated pain completed PAIN Report It Ⓡ . This computer-based instrument includes a two-dimensional, digital body outline on which patients mark their pain location. Using the ImageJ software, we calculated the percentage of the body surface area marked as painful and summarized data with descriptive statistics and a pain frequency map. The painful body areas most frequently marked were the left leg-front (73%), right leg-front (72%), upper back (72%), and lower back (70%). The frequency of pain marks in each of the 48 body segments ranged from 3 to 79 (mean, 33.2 ± 21.9). The mean percentage of painful body surface area per segment was 10.8% ± 7.5% (ranging from 1.3% to 33.1%). Patient-reported pain locations can be easily analyzed from digital drawings using an algorithm created via the free ImageJ software. This method may enhance comprehensive pain assessment, facilitating research and personalized care over time for patients with various pain conditions.

Comparing feature selection and machine learning approaches for predicting CYP2D6 methylation from genetic variation.

Introduction: Pharmacogenetics currently supports clinical decision-making on the basis of a limited number of variants in a few genes and may benefit paediatric prescribing where there is a need for more precise dosing. Integrating genomic information such as methylation into pharmacogenetic models holds the potential to improve their accuracy and consequently prescribing decisions. Cytochrome P450 2D6 (CYP2D6) is a highly polymorphic gene conventionally associated with the metabolism of commonly used drugs and endogenous substrates. We thus sought to predict epigenetic loci from single nucleotide polymorphisms (SNPs) related to CYP2D6 in children from the GUSTO cohort. Methods: Buffy coat DNA methylation was quantified using the Illumina Infinium Methylation EPIC beadchip. CpG sites associated with CYP2D6 were used as outcome variables in Linear Regression, Elastic Net and XGBoost models. We compared feature selection of SNPs from GWAS mQTLs, GTEx eQTLs and SNPs within 2 MB of the CYP2D6 gene and the impact of adding demographic data. The samples were split into training (75%) sets and test (25%) sets for validation. In Elastic Net model and XGBoost models, optimal hyperparameter search was done using 10-fold cross validation. Root Mean Square Error and R-squared values were obtained to investigate each models' performance. When GWAS was performed to determine SNPs associated with CpG sites, a total of 15 SNPs were identified where several SNPs appeared to influence multiple CpG sites. Results: Overall, Elastic Net models of genetic features appeared to perform marginally better than heritability estimates and substantially better than Linear Regression and XGBoost models. The addition of nongenetic features appeared to improve performance for some but not all feature sets and probes. The best feature set and Machine Learning (ML) approach differed substantially between CpG sites and a number of top variables were identified for each model. Discussion: The development of SNP-based prediction models for CYP2D6 CpG methylation in Singaporean children of varying ethnicities in this study has clinical application. With further validation, they may add to the set of tools available to improve precision medicine and pharmacogenetics-based dosing.

Image-based documentation of vulvodynia pain location.

Aims: Better documentation of vulvar pain is needed. We examined pain locations marked on general body and genital specific outlines among women with vulvodynia. Methods: 62 women (mean age 32.1 ± 9.5 years) with vulvodynia marked their pain on a digital genital specific outline (22 segments) and 59 of those women also marked their pain on a digital general body outline (48 segments). We used ImageJ software to determine body surface area (BSA) for each outline. Results: On the general body outline, 24/48 segments were marked; 22/22 segments were marked on the genital specific outline. There was a moderate correlation (r = 0.43; p = 0.001) between the BSA marked on the general body outline and the BSA marked on the genital area outline. Conclusions: Findings support concurrent validity of the BSA as a measure of pain location using either outline.

Women with genital pain need to be able to show their healthcare provider where they have pain, and providers need to document where women have pain. In total, 62 women with vulvodynia used a computer tablet to mark their pain on a general body outline and a genital specific outline. We used computer software to document the location of women's pain looking at percent body surface area. Women marked half of the segments on the general body outline and all the segments on the genital specific outline. Because women marked almost the same percent of body surface area on the general body outline as they did on the genital specific outline, percent body surface may be used to measure pain location.

Impact of Sensitization of Family Caregivers upon Treatment Compliance among Geriatric Patients Suffering from Elder Abuse and Neglect.

Geriatric patients in various outpatient department (OPDs) have been found to agonize from elder abuse and neglect (EAN). Such suffering imposes depressive states within individuals, which in turn affects treatment compliance. The objective of this study was to evaluate the impact of sensitization (psychotherapeutic) of family caregivers (FCGs) upon two denture treatment parameters (maintenance and treatment satisfaction) among EAN patients and compare the differences in outcome with non-abused patients. A survey of completely edentulous subjects (n = 860, aged 41-80 years) provided a sampling frame of 332 EAN patients from which 150 patients (including FCGs) fulfilling the study criteria were distributed (simple random, convenient) into two groups (Group A-control, Group B-test). FCG sensitization for subjects in Group B was performed by a clinical psychologist in 2-4 short (30 min) sessions. Demographic characteristics (frequency) were measured using a self-reported questionnaire, denture maintenance was measured using a denture hygiene index (scores), and treatment satisfaction was analyzed on a 10-point visual analog scale. Relevant data were calculated for means and absolute/relative frequencies. Any difference between two groups was estimated using an unpaired t-test while the level of relationship was determined by Karl Pearson's test at a p-value of < 0.05. The results showed highest frequency (38.6%) for neglect, with elder neglect (EN) being most common (38.14% alone and 14% in combination). EN was found more if the FCG was a son (52%), in the age group (21-30 years), and with low education and low income (75%). Patients whose FCGs were counselled (Group B) demonstrated low denture plaque scores (mean = 1.38 ± 0.618), while demonstrating comparatively higher scores in six different parameters of treatment satisfaction. Differences between the two groups for both parameters were also found to be statistically significant. Psychotherapeutic counselling in the form of FCG sensitization brings better results of denture maintenance and treatment satisfaction.

Ionic vs Manual Toothbrushes: Effect on Plaque and Oral Hygiene Status in Children.

INTRODUCTION: The foundation for healthy permanent teeth in children and teenagers is laid during the first few years of life. Toothbrushing should be presented as a habit. A wide array of toothbrushes is available in the market, which differ in size, design, length, hardness, arrangement of bristles. This present clinical study is undertaken to evaluate the effectiveness of ionic toothbrush on oral hygiene status and plaque removal. MATERIALS AND METHODS: A total of 30 children aged 6 years to 12 years were selected in this study. The children were divided into two groups, 15 children were given HyG ionic toothbrush, were instructed how to use product according to manufacturer's instructions, 15 children were assigned to use manual toothbrush, instructed to brush their teeth in usual manner. All subjects used the same commercially available dentifrice throughout the study. RESULTS: The results were evaluated at each visit day 0, day 15 and day 30 following 12-18 hours of no oral hygiene. The subjects brushed their teeth twice daily for 1 minute under supervision. The result showed that there was significant difference in all the mean values in all the parameters. It may be concluded from the present study that though the ionic toothbrush was insignificantly superior to the manual toothbrush, both the toothbrushes are clinically effective in removing plaque and improving the gingival conditions. HOW TO CITE THIS ARTICLE: Chandra S, Jain N, Garg R, et al. Ionic vs Manual Toothbrushes: Effect on Plaque and Oral Hygiene Status in Children. Int J Clin Pediatr Dent 2019;12(5):375-378.

Geriatric forensics - Part 2 "Prevalence of elder abuse and their potential forensic markers among medical and dental patients".

CONTEXT: This study is a continuation of the earlier studies and has been extended to investigate the potential forensic markers of elder abuse. AIMS: To determine the prevalence of elder abuse in various outpatient departments (OPDs). To study the associated parameters related to the abuser and the abused. To determine the existence of potential forensic markers of elder abuse. SETTINGS AND DESIGN: The subjects were randomly selected from the medical and the dental OPDs of the university. MATERIALS AND METHODS: Eight hundred and thirty two elderly subjects in the age range 40-60 years were interviewed using a questionnaire to determine the existence of elder abuse. The subjects were investigated and examined for weight, nutrition and hydration, vital signs, habits, existing visual and auditory capabilities, medications, disclosure of wills/deeds, signs of depression, and documented cleanliness. The mini-mental state examination, the Geriatric Depression Scale, the Clock drawing test, and the Brief Psychiatric Rating Scale were used to determine the potential forensic markers. STATISTICAL ANALYSIS USED: Mean values in percentage were determined by dividing the number of determined subjects by the total number of subjects for that parameter. RESULTS: About 37% in medical and 41% in dental OPDs were found to have suffered from abuse, mostly in the age group 60-70 years. Females received more abuse and a combination of son and daughter-in-law constituted most abusers. Various potential markers of elder abuse and neglect investigated among the elder abuse victims included depression (89%), signs of improper feeding (83%), changes in personal hygiene (69%), need for medical/dental treatment (78%), medication misuse (67%), changes in wills/deeds (26%), decubiti (10%), bruises (17%), skin tears (27%), and confusion (23%). CONCLUSIONS: Elder abuse exists in one or more forms in both medical and dental OPDs among both males and females in all age groups.

Priming the tooth surface with chlorhexidine and antibacterial activity of resin cement.

AIM: To evaluate the effect of priming the tooth surface with 2% chlorhexidine gluconate on antibacterial activity of resin cement. METHODS: Ten patients in whom a single missing tooth was present on both the right and left side in the upper or lower arch were selected. Two fixed partial dentures (FPDs) in each patient on the right and left side were planned. Each FPD was assigned either to the control or test group. In the control group, FPD was luted with resin cement and in the test group, the tooth surface was primed with 2% chlorhexidine gluconate before luting with resin cement. Bacteriological samples were collected at base line level, as the patient came to the outpatient department before the start of any treatment, 5 wk prior to cementation of FPD and at 13 wk (8 wk after final cementation). Microbiological processing of all samples was done and the results were statistically analyzed. RESULTS: In the test group, a predominance of aerobic/facultative gram positive cocci rod was seen which indicates a healthy periodontal site, whereas in the control group, a predominance of anaerobic gram negative rods was present which indicates an unhealthy periodontal condition. This is evident by the fact that the anaerobic bacteria percentage in the control sample is 57% and 15% in the test sample after 13 wk, whereas the aerobic/facultative bacteria percentage is 43% in the control sample and 85% in the test sample after 13 wk. The percentage of gram negative bacteria in the control sample is 61% and in the test sample is 20% after 13 wk, whereas the percentage of gram positive bacteria in the control sample is 39% and in the test sample is 80% after 13 wk. The shift from anaerobic gram negative bacteria to aerobic gram positive bacteria is clearly seen from the control to test sample after 13 wk. CONCLUSION: The present study demonstrated that priming the tooth surface with 2% chlorhexidine gluconate may enhance antibacterial activity of the resin cement.

Effect of incorporation of silane-treated silver and aluminum microparticles on strength and thermal conductivity of PMMA.

PURPOSE: The purpose of this study was to study the effect of addition of metal filler particles on different strengths of polymethylmethacrylate (PMMA) and to evaluate the thermal perception in vivo. MATERIALS AND METHODS: The study was carried out in two parts. Part 1 of the study was an in vitro investigation regarding the effect of addition of metal fillers (aluminum and silver) in concentrations of 10%, 20%, and 30%, by volume on the tensile, compressive, and flexural strength of PMMA. Part 2 of the study comprised the clinical evaluation of the thermal perception by 10 edentulous patients provided with two sets of complete dentures, one fabricated with unfilled PMMA and another with 20% aluminum particle filled PMMA on the palatal portion of the maxillary denture. Recorded data were subjected to Student's t-test and ANOVA test. RESULTS: The mean tensile and flexural strength values among control and other groups were found to have statistically significant differences (p < 0.05) except for Al1 and Al2 groups. Mean compressive strength values among control and other groups were statistically significant (p < 0.05). In the clinical study, all 10 participants reported higher perception of hot and cold sensations in dentures with a metalized palatal portion. CONCLUSIONS: Compressive strength increased progressively on increasing the filler concentration for both silver- and aluminum-filled PMMA. Silane-treated metalized PMMA showed reduction in tensile and flexural strength at 30% concentration. Metalized dentures led to an appreciable increase in thermal perception by the participants of this study.