Copper oxide nanosheets as an effective nanozyme with haloperoxidase-like activity for the colorimetric detection of H2O2 and glucose.

Copper oxide nanosheets (CuO NSs) have been successfully obtained by exploiting an effective one-step approach of sugar-blowing method followed by calcination. The nanosheets were characterized by several techniques like X-ray powder diffraction (XRD), Transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR). Impressively, CuO NSs display haloperoxidase (HPO) like catalytic activity which catalyses the oxidation of chloride ions by H2O2 giving rise to reactive chlorine species (RCS). A sensitive and selective colorimetric sensor was then demonstrated via the oxidation of chromogenic substrate 3,3',5,5'- tetramethylbenzidine (TMB) by the novel nanoenzyme CuO NSs through the generation of RCS for H2O2 and glucose detection with limit of detection of 109 nM and 21 nM in the linear ranges of 4.6 µM to 769 µM and 0.22 µM to 19.57 µM respectively. Additionally, the methodology is validated for the analysis of glucose in real samples.

CDK5 as a therapeutic tool for the treatment of Alzheimer's disease: A review.

Alzheimer's disease (AD) remains one of the most challenging and prevalent neurodegenerative disorders worldwide. Despite extensive research efforts, effective treatments for AD are lacking, emphasising the need for a deeper understanding of its underlying molecular mechanisms. Cyclin-dependent kinase 5 (CDK5), a serine/threonine kinase primarily associated with cell cycle regulation and neuronal development, has emerged as a key player in AD pathology. This review article comprehensively explores the multifaceted roles of CDK5 in the pathogenesis of AD. We begin by elucidating the physiological functions of CDK5 in normal brain development and neuronal maintenance, highlighting its involvement in synaptic plasticity, neurotransmitter release, and cytoskeletal dynamics. Subsequently, we delve into the dysregulation of CDK5 activity observed in AD, encompassing aberrant hyperactivation, and dysregulated protein interactions. Moreover, we discuss the intricate interplay between CDK5 and AD-related proteins, including amyloid-beta precursor protein (APP) and tau protein, elucidating their collective impact on disease progression. Finally, we described various approaches available for the inhibition of CDK-5, which can be explored as future therapeutic intervention for AD. Through synthesizing evidence from in vitro studies, animal models, and clinical investigations, this review provides a comprehensive overview of the intricate relationship between CDK5 dysregulation and AD pathogenesis, offering insights that may inform future therapeutic interventions strategies.

Love wave on a flexoelectric piezoelectric-viscoelastic stratified structure with dielectrically conducting imperfect interface.

The present work investigates the propagation of Love wave in a flexoelectric piezoelectric-viscoelastic stratum imperfectly bonded to a piezoelectric-viscoelastic substrate. To study the impact of imperfect interfaces, the non-traditional boundary conditions for two different types of imperfect interfaces have been taken into account. The frequency relation has been obtained in complex form employing a suitable variable separable technique. The dispersion and damping equations of Love wave have been analytically determined in the closed form by separating the real and imaginary part of the frequency relation for two distinct interfaces, i.e., mechanically compliant and dielectrically weakly/highly conducting interfaces in both electrically open and electrically short cases. Numerical calculations are carried out to reveal the impact of the width of stratum, imperfectness parameter, flexoelectric parameters, and flexoelectric loss moduli on the phase velocity and attenuation coefficient of Love wave and are also depicted through graphs for both the interfaces. For validation purposes, the expressions derived as a result of the present study are matched with the standard Love wave equation.

A Long Term Follow-Up Study of the Reconstruction of the Oral Cavity at a Tertiary Care Center: An Original Research.

Introduction: In the subcontinent of India as well as for the other nations, the most usual cancer that affects the oral cavity the "squamous cell carcinoma". The main side effects of the intervention of the SCC are the large defect and disfigurement. This study sought to investigate the various interventions of the SCC in a designated cancer institute. Material and Methods: A retrospective clinical study of the records was done between 2000-2020 years. The operated cases were analysed for various patient demographics as well as the site of the lesion and number distributions. The change in the techniques and the preference flaps over the periods was noted, and the values were compared for every 5 years. Results: A1001 patients were treated between the intended periods. Though there were larger number of men, the variation was not significant. Common sites were floor of the mouth (FOM) and tongue. The common interventions were primary closure, soft tissue free flaps and bone flaps. The shift was seen towards the free flaps. Conclusion: The Free flaps were the common applications in the constructions. Bone free flaps overtook the other procedures that were commonly applied. The quality of life and the aesthetics are seen to be better with the free flaps.

Evaluation of the Efficacy of a New Dichoptic Digital Platform to Treat the Anisometropic and Isometropic Amblyopia.

The aim of the current study was to evaluate the results of a novel dichoptic training program using an online platform in a group of subjects with refractive amblyopia, performing a comparative analysis of unilateral and bilateral amblyopic cases. For this purpose, a retrospective study analysis of data of 161 children (4−13 years) who underwent dichoptic treatment with the Bynocs® platform (Kanohi Eye Pvt. Ltd., Mumbai, India) was performed. In all cases, the therapy protocol consisted of sessions of training of 30 min daily 5 times a week for 6 weeks. Best corrected visual acuity (BCVA) in the non-dominant eye improved significantly with the treatment, with a mean change of 0.39 logMAR in the whole sample (p < 0.001). Regarding binocularity, the binocular function (BF) score also experienced a significant improvement (p < 0.001), with a mean change of 1.55 with therapy in the whole sample. The BCVA of the dominant eye only improved significantly (p < 0.001) in the isometropic amblyopic subgroup. In conclusion, the use of the dichoptic therapy with the digital platform evaluated allows an effective restoration of visual acuity and binocular function in children with anisometropic and isometropic amblyopia.

Maternal Chromium Levels in Gestational Diabetes: Systematic Review and Meta-Analysis.

The topic of maternal Chromium (Cr) levels in Gestational Diabetes Mellitus (GDM) has remained controversial; some studies have found lower levels of Cr in GDM population, whereas others found no significant changes in Cr status in GDM. Therefore, this systematic review and meta-analysis was aimed at qualitatively and quantitatively synthesizing past studies to find the relationship of maternal Cr levels with GDM. The study protocol was registered at International prospective register for systematic reviews (PROSPERO) (ID CRD42021272979). Strict adherence to the Preferred Reporting Items for Systematic review and Meta-analysis checklist, 2009 was followed during the entire study. Random-effect model for calculation of distribution of true effect sizes was used for the meta-analysis with a confidence interval (CI) of 95%. The pooled Standard Mean Difference of control and GDM groups were compared using Z statistics with a P value of <.05 as significant. Six studies were included for the systematic review and four studies entered meta-analysis. The test of overall effect revealed that the pooled Cr values did not differ significantly between controls and GDM group (Z = 1.52, P =0.13). Heterogeneity between the studies was high (I2 = 97%). A subgroup analysis revealed that results varied as per place of study, trimester of pregnancy, and Cr estimation technique. Results from meta regression analysis revealed that sample size of individual studies (Q = 0.003, P =0.67) and year of publication of studies (Q = 0.22, P =.48) had no significant effect on the overall Standard Mean Difference. Factors such as ethnicity, lack of history of infection, and diet history can influence the results of this study.

Direct detection of resistance to fluoroquinolones/SLIDs in sputum specimen by GenoType MTBDRsl v.2.0 assay A study from Eastern Uttar Pradesh, India.

BACKGROUND: According to World Health Organization (WHO), drug-resistant tuberculosis (DR-TB) is a major contributor to antimicrobial resistance globally and continues to be a public health threat. Annually, about half a million people fall ill with DR-TB globally. The gradual increase in resistance to fluoroquinolones (FQs) and second-line injectable drugs (SLIDs), poses a serious threat to effective TB control and adequate patient management. Therefore, WHO suggests the use of GenoType MTBDRsl v.2.0 assay for detection of multiple mutations associated with FQs and SLIDs. Hence, the study was conducted to determine the prevalence of resistance to FQs and SLIDs by comparing direct GenoType MTBDRsl v.2.0 assay with phenotypic drug susceptibility testing (DST). METHODS: The study was conducted on 1320 smear positive sputum samples from a total of 2536 RR-TB, confirmed by GeneXpert MTB/RIF. The smear positive specimens were decontaminated, and DNA extraction was performed. Furthermore, the extracted DNA was used for GenoType MTBDRsl v.2.0 assay. While 20% of the decontaminated specimens were inoculated in Mycobacterium growth indicator tube (MGIT) for drug susceptibility testing (DST). RESULTS: Out of 1320 smear positive sputum samples, 1178 were identified as Mycobacterium tuberculosis complex (MTBC) and remaining were negative by GenoType MTBDRsl v.2.0 assay. Of the 1178 MTBC positive, 26.6% were sensitive to both FQs and SLIDs, whereas 57.3% were only FQs resistant and 15.9% were resistant to both FQs and SLIDs. Further DST of 225 isolates by liquid culture showed that 17% were sensitive to both FQs and SLIDs, 61.3% were only FQs resistant and 21.3% were resistant to both. The specificity for FQs and SLIDs was 92.31% and 100% whereas sensitivity was 100% respectively by GenoType MTBDRsl v.2.0 assay in direct sputum samples. CONCLUSIONS: Our study clearly suggests that GenoType MTBDRsl v.2.0 assay is a reliable test for the rapid detection of resistance to second-line drugs after confirmation by GeneXpert MTB/RIF assay for RR-TB. Though, high rate FQ (ofloxacin) resistance was seen in our setting, moxifloxacin could be used as treatment option owing to very low resistance.

Evaluation of the Efficacy of Regional Anesthesia and Intramuscular Diclofenac in the Management of Postoperative Pain: A Comparative Study.

AIM: Managing postoperative pain following maxillofacial surgery is an important task. Our study aims to compare this with regional anesthesia or IM diclofenac. MATERIALS AND METHODS: This study included 30 patients who underwent bi-jaw orthognathic surgery between April 2016 and January 2020. Two groups were formed. Group A was administered inferior alveolar nerve block at the surgical site bilaterally using 0.5% ropivacaine and Group B were administered 75 mg intramuscular diclofenac just before extubation. Tramadol HCl 2 mg/kg body wt is used as a rescue analgesic. The pain was evaluated periodically at 2nd, 4th, 6th, and 12 h postoperatively. RESULTS: The mean Visual Analog Scale score was 2 in Group A and 5 in Group B. The mean duration of analgesia was 6 h 42 min, whereas in Group B, it was 8 h and 5 min. In 2 patients (13.3%) belonging to Group A and 3 patients (20%) belonging to Group B. There were no local complications in any group. CONCLUSION: It was observed that regional anesthesia in the form of intraoral nerve blocks was efficient than diclofenac (75 mg) administered intramuscularly for the management of immediate postoperative pain.

Detection of clinically important non tuberculous mycobacteria (NTM) from pulmonary samples through one-step multiplex PCR assay.

BACKGROUND: The burden of non-tuberculous mycobacterial (NTM) disease is increasing worldwide but still its diagnosis is delayed and it is mistaken as multidrug-resistant tuberculosis (MDR-TB).The present study was performed to develop a multiplex PCR assay for detection and identification of clinically most common NTM to the species level from pulmonary samples. RESULTS: Out of 50 isolates, 26 were identified as Mycobacterium kansasii (MK), 20 were identified as Mycobacterium abscessus (MA) and 4 were identified as Mycobacterium avium complex (MAC) through multiplex PCR and further confirmed by sequencing. CONCLUSION: Our study showed that multiplex PCR assay is a simple, convenient, and reliable technique for detection and differential identification of major NTM species.

Endodontic Management of Complicated Vertical Crown-Root Fracture in Deciduous Molars: Report of Two Cases.

Dental traumatic injuries most commonly occur in the primary and mixed dentition, but vertical complicated crown-root fractures are rarely seen in children. Clinical and radiographic examination of these injuries helps in accurate diagnosis and management. According to the International Association of Dental Traumatology guidelines, treatment usually involves extraction followed by placement of a space maintainer. Cases of complicated crown and root fracture in primary posterior teeth are often unnoticed by the clinician, at the time of injury. Two such patients are presented, who reported symptoms a few weeks after their accident. They were managed conservatively by initial stabilization with stainless steel crowns, followed by root canal therapy. This report highlights the need for referral to specialists and emphasizes the importance of conservative management of primary teeth to maintain functional demands.

Mycobacterium tuberculosis nucleoside diphosphate kinase shows interaction with putative ATP binding cassette (ABC) transporter, Rv1273c.

Protein-protein interactions are crucial for all biological processes. Compiling this network provides many new insights into protein function and gives directions for the development of new drugs targeted to the pathogen. Mycobacterium tuberculosis Nucleoside diphosphate kinase (Mtb Ndk) has been reported to promote survival of mycobacterium within the macrophage and contribute significantly to mycobacterium virulence. Hence, the present study was aimed to identify and characterize the interacting partner for Ndk. The in vitro experiments, pull down and far western blotting have demonstrated that Mtb Ndk interacts with Rv1273c, a probable drug ABC transporter ATP-binding protein annotated to export drugs across the membrane. This observation was further confirmed by molecular docking and dynamic simulations studies. The homology model of Rv1273c was constructed and docked with Mtb Ndk for protein-protein interaction analysis. The critical residues involved at interface of Rv1273c-Ndk interaction were identified. MDS and Principal Component analysis carried out for conformational feasibility and stability concluded that the complex between the two proteins is more stable as compared to apo proteins. Our findings would be expected to improve the dissection of protein-protein interaction network and significantly advance our understanding of tuberculosis infection.Communicated by Ramaswamy H. Sarma.

Rv1273c, an ABC transporter of Mycobacterium tuberculosis promotes mycobacterial intracellular survival within macrophages via modulating the host cell immune response.

Mycobacterium proteins, especially cell wall associated proteins, interact with host macrophage to regulate the functions and cytokine production. So, identification and characterization of such proteins is essential for understanding tuberculosis pathogenesis. The role of the ABC transporter proteins in the pathophysiology and virulence of Mycobacterium tuberculosis is not clearly understood. In the present study, Rv1273c, an ABC transporter, has been expressed in a non-pathogenic and fast growing Mycobacterium smegmatis strain to explore its role in host pathogen interactions. Over expression of Rv1273c resulted in enhanced intracellular survival in macrophage as well as modified cell wall architecture. We found altered colony morphology and cell surface properties that might be linked with remodelling of bacterial cell wall which may help in the intracellular survival of mycobacterium. However, the enhanced intracellular survival was not found to be the consequence of an increased resistance to intracellular stresses. The activation of macrophage by Rv1273c was associated with perturbed cytokine production. Pharmacological inhibition experiment and western immunoblotting suggested that this altered cytokine profile was mediated possibly by NF-kB and p38 pathway in macrophage. Overall, the present findings indicated that Rv1273c enhanced mycobacterium persistence and mediated the evasion of immune responses during infection.

Mutation in MPT64 gene influencing diagnostic accuracy of SD Bioline assay (capilia).

BACKGROUND: Success of India's TB control program relies on rapid case detection, monitoring, care and treatment of drug resistance. Patients on multidrug resistance (MDR) treatment are monitored by follow up cultures. Discordant results (culture and smear positive while capilia negative) are usually declared negative Mycobacterium tuberculosis complex (MTBC). This study was designed to understand the possible causes of discordant results. METHODS: The capilia kit was evaluated to test its utility among 4737 follow up MDR patients enrolled during a period of 1 year. A total of 889 were liquid culture positive, 3375 were negative and 473 were contaminated. Of the 889 cultures positive, 829 were found positive by ZN smear, capilia test and MTBDR plus assay. The cultures which gave a positive result on Mycobacterium Growth Indicator Tube 960 (MGIT 960) and ZN smear but were negative on capilia test with no growth on Brain Heart Infusion agar (BHI) were included in this study. The conflicting results of capilia were compared with other molecular techniques; MTBDR plus assay and DNA sequence analysis of MPT64 gene. RESULTS: Out of 889 culture positive, 60 (6.7%) were found positive on liquid culture and ZN smear but were negative on capilia. Of these 60 cultures, 10 (16.7%) were found positive by both MTBDR plus assay and PCR. The sequencing analysis revealed that all of the capilia negative isolates had mutations within the MPT64 gene. CONCLUSION: Re-evaluation of culture positive but capilia negative isolates should be done before declaring them as Mycobacterium other than tuberculosis (MOTT) because such cases can act as chronic carriers of TB in the population which can lead to the rise of this lethal disease.

SIRT6 Is a Target of Regulation by UBE3A That Contributes to Liver Tumorigenesis in an ANXA2-Dependent Manner.

UBE3A is an E3 ubiquitin ligase well known for its role in the proteasomal degradation of p53 in human papillomavirus (HPV)-associated cancers. Here we report that UBE3A ubiquitylates and triggers degradation of the tumor-suppressive sirtuin SIRT6 in hepatocellular carcinoma. UBE3A ubiquitylated the highly conserved Lys160 residue on SIRT6. FOXO1-mediated transcriptional repression of UBE3A was sufficient to stabilize SIRT6 and to epigenetically repress ANXA2, a key mediator of UBE3A oncogenic function. Thus, UBE3A-mediated SIRT6 degradation promoted the proliferative capacity, migration potential, and invasiveness of cells. In mouse models of hepatocellular carcinoma, SIRT6 downregulation and consequent induction of ANXA2 were critical for UBE3A-mediated tumorigenesis. Furthermore, in clinical specimens, increased UBE3A levels correlated with reduced SIRT6 levels and elevated ANXA2 levels in increasing tumor grades. Overall, our findings show how the tumor suppressor SIRT6 is regulated in hepatocellular carcinoma and establish the mechanism underlying UBE3A-mediated tumorigenesis in this disease.Significance: These findings provide mechanistic insights into regulation of the tumor suppressive sirtuin SIRT6 and its implications for the development of hepatocellular carcinoma. Cancer Res; 78(3); 645-58. ©2017 AACR.

Molecular detection of drug resistance to ofloxacin and kanamycin in Mycobacterium tuberculosis by using multiplex allele-specific PCR.

Drug resistance in tuberculosis (TB) is the biggest global health challenge as it hinders the tuberculosis control program and makes the disease more worsen. Molecular methods interrupt the spread of drug resistance by facilitating the appropriate anti- tuberculosis therapy at correct time through rapid diagnosis of multi drug resistant (MDR) and extensively drug resistant tuberculosis (XDR-TB). In this study we standardized and evaluated the diagnostic utility of multiplex allele specific PCR (MAS-PCR) targeting gyrA D94G and rrs A1401G mutations for detection of resistance against two key drugs (ofloxacin and kanamycin) of second line anti tuberculosis treatment. MAS-PCR assays targeting gyrA D94G and rrs A1401G for ofloxacin (OFL) and kanamycin (KAN) resistance respectively were carried out on 150 multidrug resistant isolates of Mycobacterium tuberculosis. The results were compared with phenotypic drug susceptibility test against ofloxacin and kanamycin by using proportion method on MGIT 960. Of 150 MDR isolates 50 were resistant to both ofloxacin and kanamycin, 36 were resistant to ofloxacin only, 8 were resistant to kanamycin only and 56 were susceptible to both the drugs. MAS-PCR correctly identified gyrA D94G and rrs A1401G mutations in phenotypically resistant isolates with a specificity of 100%. The sensitivity of MAS-PCR was 88.66%, 93.55% and 86% for OFL, KAN and XDR-TB respectively. There was no mutation detected at gyrA D94G region of 12.86% (11 of 86) OFL resistant isolates while 6.89% (4 of 58) of KAN resistant isolates did not carry rrs A1401G substitution. MAS-PCR proves to be a rapid tool for detection of drug resistance which could also be used as an initial marker for screening of XDR-TB.

Rapid Screening of MDR-TB in Cases of Extra Pulmonary Tuberculosis Using Geno Type MTBDRplus.

BACKGROUND: Drug resistance in tuberculosis is a major public health challenge in developing countries. The limited data available on drug resistance in extra pulmonary tuberculosis stimulated us to design our study on anti-tuberculosis drug resistance pattern in cases of extra pulmonary tuberculosis in a tertiary referral hospital of North India. We performed Geno Type MTBDRplus assay in comparison with conventional drug susceptibility testing by proportion method to study the mutation patterns in rpoB, katG and inhA genes. METHODS: A total of 510 extra pulmonary samples were included in this study. After the smear microscopy, all the specimens were subjected for culture on Lowenstein Jensen (LJ) media. Phenotypic drug susceptibility testing (DST) was performed on LJ media for all the MTB isolates and compared with the results of Geno Type MTBDRplus assay which was performed with the DNA isolated from the culture by conventional method. RESULTS: Of 510 specimens cultured, the total culture positivity obtained was 11.8% (60) encompassing 54 (10.6%) Mycobacterium tuberculosis and 6 (1.2%) non-tubercular mycobacteria (NTM). DST results by Geno Type MTBDRplus assay and solid culture methods were compared in 51 MTB isolates excluding the two Rif indeterminate and one invalid test. Geno Type MTBDRplus accurately identified 13 of 14 rifampicin-resistant strains, 14 of 15 isoniazid-resistant strains and 13 of 14 as multi drug resistant tuberculosis (MDR-TB) in comparison with conventional method. Sensitivity and specificity were 92.86% and 97.30% respectively for detection of RIF resistance, 93.33% and 94.44% respectively for detection of INH resistance, 92.86% and 97.30% respectively for detection of MDR-TB, while the overall concordance of Geno Type MTBDRplus assay with conventional DST was 94.11%. The turn-around time for performing Geno Type MTBDRplus assay test was 48 hours. CONCLUSION: The problem of MDR in extra pulmonary tuberculosis (EPTB) cannot be overlooked and due attention on patients should be given. Routine use of Geno Type MTBDRplus assay for the diagnosis of MDR-EPTB can substantially reduce the time between diagnosis and drug therapy. Culture along with Geno Type MTBDRplus assay could be a solution for rapid and accurate diagnosis of MDR-TB in low bacillary non sputum specimens.

RP - HPLC method for the estimation of Tamsulosin Hydrochloride in Tablet Dosage Form.

A rapid and sensitive reverse phase RP-HPLC method is proposed for the estimation of tamsulosin hydrochloride in tablets. Tamsulosin hydrochloride was chromatographed on a reverse phase C18 column with a mobile phase consisting of acetonitrile and water in the ratio of 50:50 v/v. The mobile phase was pumped at a flow rate of 1.5 ml/min. The eluents were monitored at 214 nm. The retention time of the drug was 1.7 min. With this method, linearity was observed between area under curve and concentration of tamsulosin hydrochloride in the injected solution, in the range of 5 to 100 µg/ml. The method was found to be applicable for analysis of the drug in tablets. The results were validated statistically.