Superoxide Anions Inhibit Intracellular Calcium Response in Porcine Airway Smooth Muscle Cells.

Background Superoxide anions (O 2 - ) have multiple effects on pulmonary parenchyma altering cell proliferation, cellular metabolism, and airway smooth muscle (ASM) contraction. Intracellular calcium ([Ca 2+ ] i ) concentration plays a significant role in the regulation of ASM contraction, relaxation, proliferation, and gene expression. Objective We investigated the effects of O 2 - on agonist-stimulated changes in [Ca 2+ ] i in ASM cells. Design/Methods Fura-2 AM-loaded, freshly isolated porcine ASM (PASM) cells were used to examine [Ca 2+ ] i release in response to acetylcholine (ACh), histamine, endothelin, caffeine, and thapsigargin (TPG) in the presence or absence of extracellular Ca 2+ . Results Exposure of PASM cells to xanthine and xanthine oxidase (X + XO) resulted in a time-dependent generation of O 2 - , inhibited by superoxide dismutase (SOD). Preincubating PASM cells with X + XO for 15- or 45-minute inhibited net [Ca 2+ ] i responses to ACh, histamine, caffeine, and TPG compared with control cells. Pretreating PASM cells with SOD for 30 minutes mitigated the inhibitory effect of X + XO treatment on ACh-induced Ca 2+ elevation suggesting role of O 2 - . X + XO treatment also inhibited caffeine- and TPG-induced Ca 2+ elevation suggesting effect of O 2 - on [Ca 2+ ] i release and reuptake mechanisms. Conclusion Superoxide attenuates [Ca 2+ ] i release, reuptake, and may interfere with physiological functions of ASM cells.

Déjerine-Roussy syndrome presenting with atypical involuntary movements.

BACKGROUND: The Déjerine-Roussy syndrome is caused by a stroke in the posterior lateral nuclei of the thalamus. It has a 17 to 18% prevalence after a stroke involving the inferior lateral thalamus. METHODS: We reported a case of a 65-year-old, diabetic and hypertensive male who presented with sudden onset of superficial hemianesthesia, allodynia, severe and paroxysmal pain on the right side of the body with choreoathetoid movements in the upperN limb, along with slured speech, and unsteadiness while walking. CONCLUSIONS: It is characterized by superficial hemianesthesia, allodynia, severe paroxysmal pain, and choreoathetoid movements in the limbs on the paralyzed side. A posterior lateral thalamic lesion can present with ataxia hemiparesis contralateral to the side of the lesion.

Knowledge, attitudes, and practices of dentists after the COVID-19 pandemic: A cross-sectional study.

Background: Dentists are at an increased risk of encountering cross-infections because of working in close proximity with oral mucosa and saliva making use of high-speed rotary instruments during treatment generating large amount of aerosols and splatter which in turn increases the probability of nosocomial spread of infections. The present study aimed to evaluate the knowledge, attitudes, and practices of dentists after the COVID-19 pandemic. Materials and Methods: The present cross-sectional study was conducted using Google Forms wherein a well-structured questionnaire composed of a total of 25 closed-ended questions was distributed among practicing dentists through online mode. Step-wise linear regression analysis was done to determine the influence of independent variables on the knowledge, attitude, and practice scores of the participants. P < 0.05 was considered statistically significant. Results: On analyzing the results, multiple linear regression analysis revealed no significant difference in the knowledge scores among the participants, while good attitude and practice scores were significantly associated with higher qualification of the participants. Another noteworthy finding in the study was the significant correlation observed between the various components of the knowledge, attitude, and practice scores among the participants (P < 0.05). Conclusion: It could be concluded from the findings of the present survey that dentists were found to have good knowledge scores about the COVID-19 pandemic. They should, however, pay more emphasis on practicing tele-triaging and proper screening and should follow various guidelines and advisories issued from time to time by the competent authorities.

Survivin Inhibition by Piperine Sensitizes Glioblastoma Cancer Stem Cells and Leads to Better Drug Response.

Glioblastoma multiforme (GBM) cancer stem cells (GSCs) are one of the strongest contributing factors to treatment resistance in GBM. Identification of biomarkers capable of directly affecting these cells within the bulk tumor is a major challenge associated with the development of new targeting strategies. In this study, we focus on understanding the potential of the multifunctional extraordinaire survivin as a biomarker for GSCs. We analyzed the expression profiles of this gene using various publicly available datasets to understand its importance in stemness and other cancer processes. The findings from these studies were further validated using human GSCs isolated from a GBM cell line. In these GSCs, survivin was inhibited using the dietary phytochemical piperine (PIP) and the subsequent effects on stemness, cancer processes and Temozolomide were investigated. In silico analysis identified survivin to be one of the most significant differentially regulated gene in GSCs, in comparison to common stemness markers. Further validation studies on the isolated GSCs showed the importance of survivin in stemness, cancer progression and therapy resistance. Taken together, our study identifies survivin as a more consistent GSC marker and also suggests the possibility of using survivin inhibitors along with standard of care drugs for better therapeutic outcomes.

PSMC1 variant causes a novel neurological syndrome.

Proteasome 26S, the eukaryotic proteasome, serves as the machinery for cellular protein degradation. It is composed of the 20S core particle and one or two 19S regulatory particles, composed of a base and a lid. To date, several human diseases have been associated with mutations within the 26S proteasome subunits; only one of them affects a base subunit. We now delineate an autosomal recessive syndrome of failure to thrive, severe developmental delay and intellectual disability, spastic tetraplegia with central hypotonia, chorea, hearing loss, micropenis and undescended testes, as well as mild elevation of liver enzymes. None of the affected individuals achieved verbal communication or ambulation. Ventriculomegaly was evident on MRI. Homozygosity mapping combined with exome sequencing revealed a disease-associated p.I328T PSMC1 variant. Protein modeling demonstrated that the PSMC1 variant is located at the highly conserved putative ATP binding and hydrolysis domain, and is suggested to interrupt a hydrophobic core within the protein. Fruit flies in which we silenced the Drosophila ortholog Rpt2 specifically in the eye exhibited an apparent phenotype that was highly rescued by the human wild-type PSMC1, yet only partly by the mutant PSMC1, proving the functional effect of the p.I328T disease-causing variant.

Safety and feasibility of posterior fossa neurosurgery in the supine position - A UK series from a large centre.

BACKGROUND: Posterior fossa surgery in the supine position remains a relatively underutilised approach, compared to the routinely performed prone, park-bench or sitting positions. This surgical approach may confer additional advantages over other modalities commonly restricted by patient co-morbidity and anaesthetic concerns. The purpose of this article is to highlight this approach as a potential viable, safe and alternative approach. METHODS: We retrospectively collected data of all supine infra-tentorial/posterior fossa surgery by one surgeon between 2015 and the present via our electronic patient record system. Demographic data alongside duration of surgery, ASA grading, location of lesions, length of stay, presence of post-operative infections, presence of post-operative CSF leak/pseudomeningocele and post-operative mortality were assessed. RESULTS: A total of 64 procedures on 58 patients were identified. Of the procedures, 60 were performed for neoplasms (93.8%). Mean overall surgical time was 176 min. Median ASA grade for tumour surgery was 3. Median length of stay was 3 days. Of the non-emergency tumour cases (n = 53), 43 (81.1%) lesions were located in the cerebellar hemisphere, the remainder were in the vermis and tentorium. There were 6 documented post-operative infections (9.4%). The rates of CSF-related complications were: CSF Leak 1.6% (1/64) and Pseudomeningocele 1.6% (1/64). 30-day mortality was 1.6% (1/64). CONCLUSION: This study suggests that supine positioning is a safe alternative to be considered when operating upon posterior fossa lesions. Further studies are warranted to assess efficacy of this approach, but it can be considered for wider use in the UK and further afield.

Rare Presentation of Limb-Body Wall Complex in a Neonate: Case Report and Review of Literature.

The limb-body wall complex (LBWC) aka body stalk syndrome is an uncommon congenital disorder characterized by severe malformations of limb, thorax, and abdomen, characterized by the presence of thoracoschisis, abdominoschisis, limb defects, and exencephaly. This condition is extremely rare with an incidence of 1 per 14,000 and 1 per 31,000 pregnancies in large epidemiologic studies. Majority of these malformed fetuses end up with spontaneous abortions. We present this rare case with occurrence in a preterm infant of 35 weeks' gestation. Our report highlights majority of the clinical presentations as reported in previous literature, but the significant pathological findings of absent genitalia and malformed genitourinary as well as anorectal malformations make this case presentation an even more rare occurrence. Infant karyotyping was normal male and there is no specific underlying genetic correlation in this condition which has a fatal prognosis.

Intratracheal Keratinocyte Growth Factor Enhances Surfactant Protein B Expression in Mechanically Ventilated Preterm Pigs.

Bronchopulmonary dysplasia (BPD) is a devastating disease of prematurity that is associated with mechanical ventilation and hyperoxia. We used preterm pigs delivered at gestational day 102 as a translational model for 26-28-week infants to test the hypothesis administering recombinant human keratinocyte growth factor (rhKGF) at initiation of mechanical ventilation will stimulate type II cell proliferation and surfactant production, mitigate ventilator induced lung injury, and reduce epithelial to mesenchymal transition considered as a precursor to BPD. Newborn preterm pigs were intubated and randomized to receive intratracheal rhKGF (20 µg/kg; n = 6) or saline (0.5 ml 0.9% saline; control; n = 6) before initiating 24 h of ventilation followed by extubation to nasal oxygen for 12 h before euthanasia and collection of lungs for histopathology and immunohistochemistry to assess expression of surfactant protein B and markers of epithelial to mesenchymal transition. rhKGF pigs required less oxygen during mechanical ventilation, had higher tidal volumes at similar peak pressures indicative of improved lung compliance, and survival was higher after extubation (83% vs. 16%). rhKGF increased surfactant protein B expression (p < 0.05) and reduced TGF-1ß (p < 0.05), that inhibits surfactant production and is a prominent marker for epithelial to mesenchymal transition. Our findings suggest intratracheal administration of rhKGF at initiation of mechanical ventilation enhances surfactant production, reduces ventilator induced lung injury, and attenuates epithelial-mesenchymal transition while improving pulmonary functions. rhKGF is a potential therapeutic strategy to mitigate pulmonary responses of preterm infants that require mechanical ventilation and thereby reduce the incidence and severity of bronchopulmonary dysplasia.

Recapitulating Actin Module Organization in the Drosophila Oocyte Reveals New Roles for Bristle-Actin-Modulating Proteins.

The generation of F-actin bundles is controlled by the action of actin-binding proteins. In Drosophila bristle development, two major actin-bundling proteins-Forked and Fascin-were identified, but still the molecular mechanism by which these actin-bundling proteins and other proteins generate bristle actin bundles is unknown. In this study, we developed a technique that allows recapitulation of bristle actin module organization using the Drosophila ovary by a combination of confocal microscopy, super-resolution structured illumination microscopy, and correlative light and electron microscope analysis. Since Forked generated a distinct ectopic network of actin bundles in the oocyte, the additive effect of two other actin-associated proteins, namely, Fascin and Javelin (Jv), was studied. We found that co-expression of Fascin and Forked demonstrated that the number of actin filaments within the actin bundles dramatically increased, and in their geometric organization, they resembled bristle-like actin bundles. On the other hand, co-expression of Jv with Forked increased the length and density of the actin bundles. When all three proteins co-expressed, the actin bundles were longer and denser, and contained a high number of actin filaments in the bundle. Thus, our results demonstrate that the Drosophila oocyte could serve as a test tube for actin bundle analysis.

Phosphoproteomics of the developing heart identifies PERM1 - An outer mitochondrial membrane protein.

Heart development relies on PTMs that control cardiomyocyte proliferation, differentiation and cardiac morphogenesis. We generated a map of phosphorylation sites during the early stages of cardiac postnatal development in mice; we quantified over 10,000 phosphorylation sites and 5000 proteins that were assigned to different pathways. Analysis of mitochondrial proteins led to the identification of PGC-1- and ERR-induced regulator in muscle 1 (PERM1), which is specifically expressed in skeletal muscle and heart tissue and associates with the outer mitochondrial membrane. We demonstrate PERM1 is subject to rapid changes mediated by the UPS through phosphorylation of its PEST motif by casein kinase 2. Ablation of Perm1 in mice results in reduced protein expression of lipin-1 accompanied by accumulation of specific phospholipid species. Isolation of Perm1-deficient mitochondria revealed significant downregulation of mitochondrial transport proteins for amino acids and carnitines, including SLC25A12/13/29/34 and CPT2. Consistently, we observed altered levels of various lipid species, amino acids, and acylcarnitines in Perm1-/- mitochondria. We conclude that the outer mitochondrial membrane protein PERM1 regulates homeostasis of lipid and amino acid metabolites in mitochondria.

An Atypical Presentation of Left Lateral Medullary Syndrome - A Case Report.

Lateral medullary syndrome (LMS), known as Wallenberg's syndrome and posterior inferior cerebellar artery syndrome, is a rare cause of stroke. It often results from thrombosis or emboli of the vertebral artery or posterior inferior cerebellar artery. The triad of Horner's syndrome, ipsilateral ataxia, and ipsilateral hyperalgesia clinically identify patients with LMS. We report a case of a 62-year-old diabetic, hypertensive male who presented with symptoms involving the left lateral dorsal medulla along with partial Horner's syndrome, left lateral rectus palsy, and left lower motor neuron-type facial palsy. Atypical presentation in LMS could be explained by infraction of left facial colliculus in addition to the left lateral medulla. Association of these entities with Wallenberg's syndrome is atypical features in our case, which questioned the diagnosis of a simple LMS.

Revisiting the Role of ß-Tubulin in Drosophila Development: ß-tubulin60D is not an Essential Gene, and its Novel Pin 1 Allele has a Tissue-Specific Dominant-Negative Impact.

Diversity in cytoskeleton organization and function may be achieved through alternative tubulin isotypes and by a variety of post-translational modifications. The Drosophila genome contains five different ß-tubulin paralogs, which may play an isotype tissue-specific function in vivo. One of these genes, the ß-tubulin60D gene, which is expressed in a tissue-specific manner, was found to be essential for fly viability and fertility. To further understand the role of the ß-tubulin60D gene, we generated new ß-tubulin60D null alleles (ß-tubulin60D M ) using the CRISPR/Cas9 system and found that the homozygous flies were viable and fertile. Moreover, using a combination of genetic complementation tests, rescue experiments, and cell biology analyses, we identified Pin 1 , an unknown dominant mutant with bristle developmental defects, as a dominant-negative allele of ß-tubulin60D. We also found a missense mutation in the Pin1 mutant that results in an amino acid replacement from the highly conserved glutamate at position 75 to lysine (E75K). Analyzing the ß-tubulin structure suggests that this E75K alteration destabilizes the alpha-helix structure and may also alter the GTP-Mg2+ complex binding capabilities. Our results revisited the credence that ß-tubulin60D is required for fly viability and revealed for the first time in Drosophila, a novel dominant-negative function of missense ß-tubulin60D mutation in bristle morphogenesis.

A Comparative Study to Evaluate the Effects of Probiotic Curd on Streptococcus mutans, Bifidobacterium dentium, and pH of Saliva in Caries-free Children: An In Vivo Study.

BACKGROUND: Dental caries is one of the commonest oral diseases that require early stages of prevention and intervention. The primary organisms related to dental caries are streptococcus mutans and lactobacilli. In the past decades, some of the non-pathogenic bacteria called probiotics have been added to the food products for the beneficial effect of human health. Intake of probiotic food products has been shown to have beneficial effects on the oral tissues. Hence the present study aimed to compare the levels of salivary pH, S. mutans, B. dentium counts in children consuming probiotic and non-probiotic curd. METHODS AND MATERIAL: 40 healthy caries free children were randomly allocated into two groups. First baseline salivary samples were collected from all the participants and tested for salivary pH, salivary B. dentium and S. mutans count. After collecting the baseline values test group of children were provided with probiotic curd whereas the control group were provided with non probiotic curd for 15 days. After 15 days of consumption of these dairy products, salivary samples were collected again and tested for salivary pH, salivary B. dentium and S. mutans count. RESULTS: Probiotic curd was found to be more effective in reducing the colony counts of B.dentium and S.mutans. There was a marginal decrease in pH in both the groups but was well above the critical pH. CONCLUSIONS: Probiotic curd showed considerable level of decrease in colony counts of B.dentium, S.mutans and salivary pH when compared to normal curd.

Perception, Knowledge, and Attitude of Problem-based Learning among Dental College Students in India: A Closed-ended Questionnaire Study.

CONTEXT: Problem-based learning (PBL) is mostly a student-centered learning method, and it is a new method used in medical and dental education all over the world. In India, PBL is a part of the learning process sparingly followed. Contrary to other institutions, PBL has been introduced in dental curriculum along with the traditional curriculum very recently. AIM: The aim of this study was to evaluate the newly introduced PBL curriculum among the dental students using a closed-ended questionnaire on a five-point scale. MATERIALS AND METHODS: The questionnaire on PBL was prepared in Google Forms, comprising 22 closed-ended questions on a five-point Likert scale, ranging from strongly disagree (1) to strongly agree (5). We carried out a pilot study in 10% of the target population to check validity and feasibility. Final study was conducted in the targeted population of 185 dental students who had undergone PBL sessions. RESULTS: The responses for the questionnaire thus received from the students were compiled and analyzed using Google Forms software. Overall rating of PBL by the respondents was as follows. The majority of the student's responses were agreed with 54.6% and a significant number of students strongly agreed with PBL at 33%. CONCLUSIONS: This study shows the responses of the students on aspects of group learning, imparting content knowledge, improvement of group skills, improvement of communication skills, and carrying out self-directed learning within the Indian context of dental health education.

Efficacy of a dementia intensive support (DIS) service at preventing admissions to medical and psychiatric wards: qualitative and quantitative evaluation.

AIMS AND METHOD: To establish whether a dementia intensive support (DIS) service that is part of a crisis resolution and home treatment team for older people is preventing admissions to acute hospital and psychiatric wards. The number of referrals in 2017 to the DIS service was established and those admitted to hospital ascertained. Senior doctors examined 30 sets of notes in detail and reached a conclusion on whether DIS had contributed to admission prevention. This information was then re-examined in two meetings with at least eight senior psychiatrists present. A consensus opinion was then reached as to whether DIS had contributed to admission prevention in each case. RESULTS: Over 12 months, 30/171 patients (18%) referred were admitted to hospital. For the subset of 30 referrals examined in detail, DIS contributed to admission avoidance in 21 cases (70%). CLINICAL IMPLICATIONS: Our evaluation demonstrates that the DIS service is an effective way of preventing admission.

Evaluation of Postsurgical Dentofacial Deformities in Children Operated for Correction of Cleft Lip and Palate-A Cross-sectional Study.

AIM: The aim of this study is to evaluate the various dentofacial deformities in children who were surgically managed for cleft lip and palate (CLP). MATERIALS AND METHODS: The study was carried out in 100 surgically managed cleft lip palate children between the ages of 4 years and 15 years. After eliciting a detailed history, a thorough intraoral and extraoral examination was done and details were recorded in a printed proforma with photographs. DESIGN: Cross-sectional study. STATISTICAL ANALYSIS USED: All the data were analyzed using SPSS 11.5 software for evaluation using the Chi-square test. RESULTS: A wide range of surgical, dental, and functional problems among the surgically managed CLP patients were seen. CONCLUSION: The timing of direct lip repair showed a significant influence on the severity of dentofacial deformities. Lip repair before the age of 1 increases the severity of the deformity. HOW TO CITE THIS ARTICLE: Joybell C, Krishnan R, et al. Evaluation of Postsurgical Dentofacial Deformities in Children Operated for Correction of Cleft Lip and Palate-A Cross-sectional Study. Int J Clin Pediatr Dent 2019;12(3):165-177.

Management and Disposal of Mercury and Amalgam in the Dental Clinics of South India: A Cross-Sectional Study.

CONTEXT: Dental offices are known to be one of the largest users of inorganic mercury in the preparation of amalgam-a restorative material which, if not handled and disposed through scientific methods, can pose grave threats to the biosphere. AIMS: The objective of this study was to assess and record the mercury management and disposal strategies of dental practitioners belonging to the two South Indian states, Kerala and Tamil Nadu. SUBJECTS AND METHODS: A questionnaire regarding the usage and disposal of a filling material containing mercury (amalgam) was designed and distributed online. The 150 dental practitioners partaking in this study responded anonymously. RESULTS: The results were statistically analyzed using chi-square test and the P value was evaluated. The usage of amalgam was correlated with the age of the practitioner and the nature of practice and it was found to be statistically significant. The number of fillings was correlated with the years of practice and locality of the clinic, which were found to be statistically significant. CONCLUSIONS: This study showed that mercury was widely preferred and utilized even today as a restorative material by dentists and dental specialists, despite the availability of other alternative strategies. Only a minor section of practitioners were found to be aware of the global changes in the guidelines pertaining to the handling and disposal of amalgam. We feel that Safe Mercury Amalgam Removal Technique, amalgam safety rules, and amalgam-free practice should be a part of the academic curriculum and continuing dental education.

Effect of Dental Health Education on the Knowledge and Attitude Among Expectant Mothers: A Questionnaire Study.

BACKGROUND: Although dental caries levels have declined and stabilized the world over, the problem of early childhood caries has remained persistent in many areas of the world affecting certain segments of society, especially the socially deprived, who remain at high risk to this disease. Parents have an essential role in mediating between their children and environment for which prenatal period is the best time for health interventions. This mediation is done through preventive services they provide to their children such as dietary habits supervision, oral hygiene performance, and other practices that can be enhanced by prenatal dental health education. PURPOSE: The purpose of the study was to evaluate, by means of a questionnaire, the expectant mother's knowledge and attitude toward dental health and to test the effectiveness of the health education tool. MATERIALS AND METHODS: The baseline questionnaire consisted of 27 questions, which was distributed to 138 participants. Subsequently, provision for dental health education was made in batches of 20. After 3 weeks, 10 questions were randomly selected from the first questionnaire for reassessment session. Paired t test was used for statistical analysis. RESULTS: The mean overall knowledge improved from 67.4% to 97.32% and attitude improved from 75.83% to 99.22%. CONCLUSION: Knowledge and attitude of mothers was found to increase on provision of dental health education, which were retained even after a period of 3 weeks.

Correlation of ICDAS-LAA and Ora test for Caries Risk Assessment: A Cross-sectional Study.

BACKGROUND: Dental caries is the most common chronic disease of children aged 6-11 years. The International Caries Detection and Assessment System-Lesion Activity Assessment (ICDAS-LAA) is a precise tool for the clinical assessment of dental caries by visual appearance, tactile feeling, and potential for plaque accumulation. Ora test is a chairside tool to estimate the ability of acid produced by bacteria to cause demineralization, in a suitable environment. Aim of this study is to correlate ICDAS-LAA and Ora test in assessing the caries risk of a patient on the dental chair. MATERIALS AND METHODS: A total of 60 school children aged between 6 and 9 years were included in the study. Of 60 children, 30 (Group A) were caries free and 30 (Group B) had decayed missing extraction, filled (DMF) score of 1-3. All the children were initially subjected to ICDAS-LAA scoring, which was assessed by one examiner followed by chairside Ora test performed by another examiner. RESULTS: Statistical analysis was done using Pearson correlation test, and a negative linear relation was found between the time taken for color change in Ora test and ICDAS-LAA scores. CONCLUSION: A direct correlation exists between ICDAS-LAA scores and Ora test, which shows that microbial activity correlates well with lesion activity.

A rare occurrence of supplementary maxillary lateral incisors and a detailed review on supernumerary teeth.

This article highlights a rare occurrence of nonsyndromic supplemental maxillary lateral incisors in a 11-year-old boy. It also includes a detailed literature review of supernumerary teeth.