Exposure of calcium carbide induces apoptosis in mammalian fibroblast L929 cells.

Inspite of various health warnings from Government and health organizations, Calcium carbide (CaC2) is still the most commonly and widely used artificial fruit ripener, probably due to its easy availability, low cost and convenience of usage. Assessment of the hazardous effects of the CaC2 applications for fruit ripening has been a matter of interest since long. Several in vivo studies have reported the toxicological outcomes such as histopathological changes in lungs and kidneys, haematological and immunological responses, upon exposure with CaC2. However, a well-controlled study investigating the effects of CaC2 under in-vitro setup was lacking. Hence, this study has been conducted to explore the toxicity associated cellular events in L929 cells exposed with varying concentrations of CaC2 (0.00312-0.2 µg/µl) for 24 h exposure time. A 23.14% reduction in cell viability was observed at the highest dose of CaC2. A similar trend in cellular stress levels at 0.2 µg/µl dose was observed in terms of rounded cellular morphology and decreased adherence as compared to the control. Furthermore, Annexin V FITC/PI staining and subsequent confocal imaging revealed a similar trend of CaC2 induced apoptosis in a dose dependent manner. A gradual elevation of intracellular ROS has also been observed up to 0.025 µg/µl dose. Thus, the study concludes that short term CaC2 exposure may increase the cellular oxidative stress and disturb the redox balance of the cell which then undergoes apoptosis. The study concludes that the exposure of CaC2 can be associated with severe diseases and suggests to stop the uses of CaC2 as fruit ripening agent.

Computed Tomography (CT) and Magnetic Resonance (MR) Findings in Xanthogranulomatous Cholecystitis: Retrospective Analysis of Pathologically Proven 30 Cases - Tertiary Care Experience.

BACKGROUND: To study CT and MR findings in xanthogranulomatous cholecystitis (XGC). MATERIAL/METHODS: Retrospective analysis of 30 histopathologically confirmed cases of XGC. Seventeen patients underwent CECT and 13 underwent MRI. The following features were studied - wall thickness, intramural nodules, pericholecystic stranding, wall thickness, THAD, fat in gallbladder wall, cholelithiasis, infiltration, biliary dilatation, lymph nodes, complications. RESULTS: The majority of cases (22/30) showed discontinuous mucosal lining. Discontinuous mucosal lining was seen in all cases with wall thickness >10 mm, 75% of cases with wall thickness between 3-10 mm and none in normal wall thickness (p=0.03). Diffuse wall thickening was seen in 23 cases, focal thickening in 3 and polypoidal wall thickening in 2 cases. Polypoidal thickening was seen in gallbladder carcinoma. Intramural nodules were present in 87.5% of cases with discontinuous mucosal lining. Pericholecystic stranding was seen in 19, biliary dilatation in 12, liver infiltration in 13 and fat in 7 cases. Lymphadenopathy was seen in 1 case with gallbladder carcinoma. Four cases showed a signal drop in the intramural nodules on chemical shift MRI. CONCLUSIONS: Discontinuous mucosal lining is evident in xanthogranulomatous cholecystitis. Diffuse wall thickening, intramural nodules, continuous or discontinuous mucosal lining and cholelithiasis may indicate XGC rather than gallbladder carcinoma. Based on correlation with pathophysiological findings, we conclude that discontinuous mucosal lining is not an unusual finding in cases of XGC. Advances in knowledge: Being aware of the radiological findings described in this article may be helpful in making preoperative radiological diagnosis of XGC. Mucosal lining may be continuous or discontinuous in XGC.

Comparing the Effects of Virtual Reality Breathing Exercise and Incentive Spirometry Exercise on Improving Pulmonary Function in Children with Spastic Diplegic Cerebral Palsy.

INTRODUCTION: Children with cerebral palsy (CP) have a higher incidence of respiratory dysfunction than healthy children. Virtual reality breathing therapy is an assistive technology that is becoming popular in the rehabilitation of children with CP. METHODS: This experimental study included a total of 32 children with spastic diplegic CP who were divided into two groups: the virtual reality breathing training (VRBT) group and the incentive spirometry (IST) group. Individuals classified as levels I to III on the gross motor function classification system (GMFCS) were recruited using the simple random sampling method. RESULT: The results of comparing the values of forced vital capacity (FVC), forced expiratory volume at one second (FEV1), and the ratio of FVC/FEV1 showed a significant difference between groups. A significant difference was found in the VRBT group compared to the IST group, except for the peak expiratory flow (PEF) values, which showed a nonsignificant difference between the groups. CONCLUSION: There were significant differences in FVC and FEV1 between the VRBT and IST groups. It has been concluded that VRBT has additional benefits in improving pulmonary functions.

Antioxidant and Anti-inflammatory Properties of the Two Varieties of Musa acuminata: An In Vitro Study.

Background Free radicals are involved in the process of carcinogenesis. Conventional antioxidants and anti-inflammatory drugs have the disadvantages of side effects and high costs. Banana peel contains phenolic and non-phenolic antioxidants that are pivotal in removing inflammatory components by inhibiting reactive oxygen species (ROS), protecting protease inhibitors from oxidative damage, and preventing fibroblast degradation which protects the body against the ill effects of free radicals. Aim and objectives The present study aimed to evaluate the potential antioxidant and anti-inflammatory activity of peel extracts of the Musa acuminata Red Dacca(red banana) and Musa acuminata Colla (rasthali). Materials and methods The procured unripe peels of red bananas and rasthali bananas were dried, ground into powder, and used to create aqueous and alcoholic extracts. The aqueous extract was made by dissolving 5 grams of peel powder in 25 ml of distilled water, while the alcoholic extract was prepared by heating ethanol to 100°C for 30 minutes. The extracts were combined, shaken for 24 hours, filtered, and stored at 4°C. Following extract preparation, 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, hydrogen peroxide (H2O2) assay, bovine serum albumin (BSA) denaturation assay, and egg albumin (EA) denaturation assay were performed to evaluate the antioxidant and anti-inflammatory properties. The assays were performed in varying concentrations for the prepared extracts of red banana and rasthali and the 1:1 ratio combination extract of both varieties. The obtained data were tabulated and statistically tested using the IBM SPSS Statistics for Windows, Version 22.0 (Released 2013; IBM Corp., Armonk, New York, United States) by the Kruskal-Wallis test with the statistical significance set at p≤0.05. Results Results highlighted variations in the antioxidant and anti-inflammatory properties of the banana peel extracts and the standard used in all the assays, but there was no statistically significant difference between the extracts and the standard (p>0.05). There was an increase in the antioxidant and anti-inflammatory activity with an increase in the concentration of both the extracts and the standard. The 1:1 ratio combination extract showed the highest antioxidant property among the banana extracts in the majority of the concentrations in the DPPH assay, whereas the rasthali extract showed the same even more than the standard in the H2O2 assay. The rasthali extract showed the highest anti-inflammatory property in all the concentrations in the BSA assay, and the 1:1 ratio combination extract showed the same in the EA assay. Conclusion The banana peel extracts showed comparable antioxidant and anti-inflammatory properties with that of the standard in all the assays with no statistically significant difference. There was a rising trend in the properties with an increase in their concentration. Red banana and rasthali peel extracts, either individually or in combination, could be a promising, effective, and cost-effective alternative or adjunct to the currently available antioxidant medications.

Antimicrobial properties of green synthesized silver and chitosan nanocomposites.

An eco-friendly and simple approach was carried out for the synthesis of silver-chitosan nanocomposites using Azadirachta indica and fluconazole-mediated aqueous extract. This extract acted as a reducing agent as well as a capping agent for the green synthesis of silver nanoparticles. Chitosan nanoparticles on the other hand were synthesized from the deacetylation of the chitin matrix. To confirm the nanoparticle synthesis, a UV- A visible spectrophotometer was used and FTIR analysis confirmed the presence of functional groups in the prepared extract. The morphological characteristics of silver and chitosan nanoparticles and as nano-composites were studied and confirmed using scanning electron microscopy (SEM) analysis. The synthesized silver-chitosan nanocomposites were subjected to well-loaded agar plates for the evaluation of antibacterial properties against the Streptococcus mutans and Candida albicans for their antifungal properties. The synthesized silver and chitosan nanoparticles showed antibacterial and antifungal activities against common oral micro flora such as Streptococcus mutans and Candida albicans which were measured using the zone of inhibition method. This approach is a one-step, economical and eco-friendly, biocompatible, and effective alternative for nanoparticle synthesis for various prosthetic applications.

Bridging the Gap with Nanoparticles: A Novel Approach.

Two-piece implants unavoidably present a microgap between the implant and the abutment interface. Although numerous modalities have been attempted to overcome this situation, the implant abutment interface still remains a critical point for microbial colonization, which starts an inflammatory cascade of events eventually compromising the implants. Throughout our life, cells in all biological systems are unprotected to oxidative stress leading to the formation of Reactive oxygen species which is of concern when it comes to placing implants in patients who are periodontally compromised. This necessitates the development of alternative therapeutic modalities, which could counteract as well as prevent the microbial overload and ROS generation thereby improving the longevity of implants. To evaluate and assess the antibacterial, antioxidant and anti inflammatory effectiveness of quercetin-loaded titanium nanocomposites as coatings over healing abutments. Quercetin-loaded titanium nanocomposites were synthesized using green synthesis and confirmation was done using UV spectroscopy. Healing abutments were coated with the formulated nanocomposites, an intra-oral environment was simulated by thermocycling. Their antibacterial, antioxidant, anti-inflammatory, and cytotoxicity were assessed using standard tests. Healing abutments were coated with the formulated nanocomposites, an intra-oral environment was simulated by thermocycling. They showed potent antibacterial, antioxidant, and anti-inflammatory properties, which could prove beneficial in a variety of clinical scenarios in which there is a high risk for implant failure during early osseointegration.

COVID-19: Invasion, pathogenesis and possible cure - A review.

Today, Coronavirus disease (COVID-19) which is believed to be transmitted from bats to humans where the people of Wuhan city, China exposed to the wet animal market is an important international public health anxiety (Xiong et al., 2020). Although, several measures were undertaken to treat the diseases by various medical advancements and by a variety of treatment procedures, still the mortality is higher. Hence, social distancing has been implemented to control the current outburst of this pandemic which spreads through human to human transmission. As a consequence, there is a need to completely understand the route of invasions of the virus into the humans and the target receptors besides the other factors leading to the disease. Several vaccines and drugs have been developed with its own pros and cons. Many are still under the various phase of R&D and clinical trials. Here we highlight the possible entry molecules, pathogenesis, symptomatology, probable cure and the recently developed vaccines for the existing pandemic due to the COVID-19.

Green synthesis of calcium hydroxide-coated silver nanoparticles using Andrographis paniculata and Ocimum sanctum Linn. leaf extracts: An antimicrobial and cytotoxic activity.

Context: Silver is known for its antibacterial properties since ages. As nanoparticles have smaller size and greater surface area, silver has been utilized in the form of nanoparticles to enhance its antibacterial properties. Calcium hydroxide is a well-known intracanal medicament and serves as a gold standard for root canal disinfection. Using herbal extracts as reducing agents for nanoparticle synthesis appears to be an ecofriendly approach. Aim: The aim of this study was to synthesize calcium hydroxide-based silver nanoparticles using herbs as reducing agents and to test the cytotoxic levels and antimicrobial activity against oral microbes. Materials and Methods: The calcium hydroxide-based silver nanoparticles were synthesized using the leaves of Andrographis paniculata and Ocimum sanctum Linn. Various properties of the synthesized nanoparticles were also characterized by ultraviolet (UV) spectrophotometer analysis, transmission electron microscopy (TEM), X-ray diffraction (XRD), and Fourier-transform infrared spectroscopy (FTIR) analysis. The cytotoxic effects of these nanoparticles were analyzed using brine shrimp and MTT assay. Antimicrobial activity was assessed by measuring the zone of inhibition. The statistical analysis was done using parametric independent t-test. P value was set at < 0.05. Results: The calcium hydroxide-based silver nanoparticles were successfully synthesized and were confirmed by UV spectrophotometer analysis, TEM, X-ray diffraction (XRD), and FTIR analysis and showed a minimal cytotoxic effect. They also showed a good antimicrobial activity and a remarkable antifungal activity. Conclusions: The green synthesis of CaOHAgNPs yielded an effective nanoparticle preparation that could be used against common oral pathogens as a potential therapeutic agent in the form of root canal irrigant or intracanal medicament in the field of dentistry.

An Update on Advances in COVID-19 Laboratory Diagnosis and Testing Guidelines in India.

The declaration of COVID-19 as a global pandemic has warranted the urgent need for technologies and tools to be deployed for confirming diagnosis of suspected cases. Diagnostic testing for COVID-19 is critical for understanding epidemiology, contract-tracing, case management, and to repress the transmission of the SARS-CoV-2. Currently, the Nucleic Acid Amplification Test (NAAT)-based RT-PCR technique is a gold standard test used for routine diagnosis of COVID-19 infection. While there are many commercially available RT-PCR assay kits available in the market, selection of highly sensitive, specific, and validated assays is most crucial for the accurate diagnosis of COVID-19 infection. Laboratory diagnosis of SARS-CoV-2 is extremely important in the disease and outbreak management. Development of rapid point of care tests with better sensitivity and specificity is the critical need of the hour as this will help accurate diagnosis and aid in containing the spread of SARS-CoV-2 infection. Early detection of viral infection greatly enhances implementation of specific public health intervention, such as infection control, environmental decontamination, and the closure of specific high-risk zones. Large-scale sequencing of SARS-CoV-2 genome isolated from affected populations across the world needs to be carried to monitor mutations that might affect performance of molecular tests. Creation of genome repositories and open-source genetic databases for use by global researchers is clearly the way forward to manage COVID-19 outbreak and accelerate vaccine development. This review summarizes various molecular diagnostics methods, technical guidelines, and advanced testing strategies adopted in India for laboratory diagnosis of COVID-19.

Development and characterization of cell lines derived from rohu, Labeo rohita (Hamilton), and catla, Catla catla (Hamilton).

Two new cell lines, designated RE and CB, were derived from the eye of rohu, Labeo rohita, and the brain of catla, Catla catla, respectively. The cell lines were maintained in Leibovitz's L-15 supplemented with 20% foetal bovine serum. The RE cell line was sub-cultured for more than 70 passages and the CB cell line for more than 35 passages. The RE cells are rounded and consist predominantly of epithelial cells. The CB cell line consists of predominantly fibroblastic-like cells. Both cell lines are able to grow at temperatures between 25 and 32 degrees C with an optimum of 28 degrees C. The growth rate of the cells increased as the foetal bovine serum concentration increased from 2% to 20% at 28 degrees C, with optimum growth at concentrations of 15% or 20% foetal bovine serum. The cells were successfully cryopreserved and revived at different passage levels. The cell lines were not susceptible to four marine fish viruses. Extracellular products from Aeromonas sp. were toxic to the cell lines. When the cells were transfected with plasmid eukaryotic green fluorescent protein (pEGFP [Clontech, Carlsbad, CA, USA]) vector DNA, a significant fluorescent signal was observed suggesting that these cell lines could be a useful tool for transgenic and genetic manipulation studies. Polymerase chain reaction amplification of mitochondrial 12S rRNA from rohu and catla confirmed that the cell lines originated from these fish species. The cell lines were further characterized by immunocytochemistry using confocal laser scanning microscopy.

Influence of estrous stages on electrocardiography, clinical pathology and ovarian weight of experimental beagle dogs: a retrospective analysis.

Estrous cycle is a repetitive phenomenon occurring during the reproductive life of a female dog. The duration of the canine estrous cycle is considerably longer than one in the most of the other animals and is broadly grouped into follicular phase (proestrus and estrus), luteal phase (diestrus) and non-seasonal anestrus. Dogs in the same stage of cycle can be inadvertently assigned to same group during routine safety and metabolic studies leading to possible erroneous interpretation of test-item related effects. This retrospective analysis was conducted by analyzing data of 86 female beagle dogs from control/placebo treated groups to correlate any possible effect of estrous stages with electrocardiography, clinical pathology and ovarian weight. Different estrous cycle stages of beagles were confirmed histologically by evaluating ovary, uterus, vagina and mammary glands. The incidence of beagles in diestrus was the highest, followed by anestrus, proestrus and estrus. No significant effect was noticed on heart rate, P-A, P-D, RR, QRS and QT intervals across different stages of estrous cycle. However, significantly higher PQ (PR) interval in dogs in proestrus stage was observed compared to dogs in anestrus and estrus. Marginally higher WBCs, neutrophils, lymphocytes, RBCs, hemoglobin, AST and lower hematocrit, lipid profile (total cholesterol, HDL, LDL, triglycerides), ALP level was evident in estrous period. Relative ovary weight was significantly higher in dogs in diestrus stage. Considering these results, one may need to exercise caution while interpreting experimental data from female beagle dogs.

Oligonucleotide therapy: An emerging focus area for drug delivery in chronic inflammatory respiratory diseases.

Oligonucleotide-based therapies are advanced novel interventions used in the management of various respiratory diseases such as asthma and Chronic Obstructive Pulmonary Disease (COPD). These agents primarily act by gene silencing or RNA interference. Better methodologies and techniques are the need of the hour that can deliver these agents to tissues and cells in a target specific manner by which their maximum potential can be reached in the management of chronic inflammatory diseases. Nanoparticles play an important role in the target-specific delivery of drugs. In addition, oligonucleotides also are extensively used for gene transfer in the form of polymeric, liposomal and inorganic carrier materials. Therefore, the current review focuses on various novel dosage forms like nanoparticles, liposomes that can be used efficiently for the delivery of various oligonucleotides such as siRNA and miRNA. We also discuss the future perspectives and targets for oligonucleotides in the management of respiratory diseases.

Silencing VP28 gene of white spot syndrome virus of shrimp by bacterially expressed dsRNA.

An in vivo expression system to produce large amounts of virus-derived dsRNAs in bacteria to provide a practical control of white spot syndrome virus (WSSV) in shrimp was developed. The bacterially synthesized dsRNA specific to VP28 gene of WSSV promoted gene-specific interference with the WSSV infection in shrimp. Virus infectivity was significantly reduced in WSSV-challenged shrimp injected with VP28-dsRNA and 100% survival was recorded. The inhibition of the expression of WSSV VP28 gene in experimentally challenged animals by VP28-dsRNA was confirmed by RT-PCR and Western blot analyses. Furthermore, we have demonstrated the efficacy of bacterially expressed VP28-dsRNA to silence VP28 gene expression in SISK cell line transfected with eukaryotic expression vector (pcDNA3.1) inserted with VP28 gene of WSSV. The expression level of VP28 gene in SISK cells was determined by fluorescent microscopy and ELISA. The results showed that the expression was significantly reduced in cells transfected with VP28dsRNA, whereas the cells transected with pcDNA-VP28 alone showed higher expression. The in vivo production of dsRNA using prokaryotic expression system could be an alternative to in vitro method for large-scale production of dsRNA corresponding to VP28 gene of WSSV for practical application to control the WSSV in shrimp farming.

Natural aquatic insect carriers of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV).

Five different species of aquatic insects were collected from nursery ponds containing the freshwater prawn Macrobrachium rosenbergii infected with Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV). The insects were screened as potential natural carriers of MrNV and XSV. RT-PCR (reverse transcription polymerase chain reaction) analysis gave positive results for MrNV and XSV in Belostoma sp., Aesohna sp., Cybister sp. and Notonecta sp., and negative results for Nepa sp. An Aedes albopictus mosquito cell line (C6/36) was used for infectivity assays, with viral inoculum prepared from the aquatic insects, since C6/36 cells have recently been shown to be susceptible to infection with MrNV and XSV. The C6/36 cells were harvested 4 d post-challenge for examination by electron microscopy. This revealed aggregation of viral particles throughout the cytoplasm for cells challenged with inocula from all the insect species except Nepa sp. Our results indicate that several aquatic insect species may present a risk for MrNV and XSV transmission to M. rosenbergii.

Immunization with baculovirus displayed H6 hemagglutinin vaccine protects mice against lethal H6 influenza virus challenge.

Low pathogenic influenza viruses of H6 hemagglutinin (HA) subtype have a high prevalence among aquatic and domestic birds and have caused outbreaks in poultry worldwide. The first human infection with wild avian influenza H6N1 virus was reported in Taiwan and these subtype viruses may continue to evolve and accumulate changes which increasing the potential risk of human-to-human transmission. To develop a vaccine against influenza viruses of the H6 subtype, we displayed the HA gene on the baculovirus surface (Bac-HA), and studied its vaccine efficacy against a lethal challenge with mouse-adapted RG-H6(Shorebird) virus carrying the H6 HA gene from A/shorebird/DE/12/2004 (H6N8) virus and 7 genes from A/Puerto Rico/8/1934 (H1N1) virus. Immunization with 256 HA units of Bac-HA via the intranasal route triggered HA-specific serum and mucosal antibodies in mice besides increased HA inhibition titers compared to mice immunized subcutaneously. Moreover, we observed an increase in cellular immune response (IL-4) and improved in vitro neutralization activity in the mice immunized intranasally with live Bac-HA compared to mice immunized with inactivated influenza virus (IV). Interestingly, Bac-HA intranasal immunized mice showed one fold higher neutralization titer against heterologous H6 influenza virus compared to inactivated IV immunized mice. In addition, the live Bac-HA, administered through either immunization route, as well as the adjuvanted inactivated Bac-HA, administered subcutaneously, conferred 100% protection to mice challenged with homologous mouse-adapted RG-H6(Shorebird) virus. The reduction in viral titers and extend of histopathological changes of Bac-HA immunized mice lungs further demonstrated the protective efficacy of Bac-HA. Hence, the recombinant baculovirus subunit vaccine is an alternative candidate against H6 subtypes that could be propagated and administered with minimal biosafety concerns.

Quercetin conjugated superparamagnetic magnetite nanoparticles for in-vitro analysis of breast cancer cell lines for chemotherapy applications.

The magnetic nanoparticles attract increasing interest due to their opportunities in cancer therapy and used as drug carriers for several other diseases. The present study investigates the quercetin conjugated superparamagnetic Fe3O4 nanoparticles for in-vitro analysis of breast cancer cell lines for chemotherapy. A simple precipitation method was used to prepare the dextran coated Fe3O4 nanoparticles and the anticancer flavonoid quercetin was conjugated on the surface via carboxylic/amine group using nanoprecipitation method. The structural, morphological and the magnetic properties of the prepared materials were studied by using X-ray diffractometer (XRD), Fourier transformed infer-red spectrometer (FTIR), transmission electron microscope (TEM) and vibrating sample magnetometer (VSM). The MTT (3-(4,5-dimethylthiahiazol-2-yl)-2,5-diphenyl tetrazolium) assay of dextran coated Fe3O4 nanoparticles did not exhibit notable toxicity against MCF7 cells, whereas the cytotoxicity of quercetin conjugated Fe3O4 nanoparticles increased significantly in comparison with pure quercetin. The incubation of MCF-7 cells with quercetin conjugated Fe3O4 nanoparticles (QCMNPs) shows significant changes in cellular morphology observed through fluorescent microscopy. The results validate the prepared quercetin conjugated Fe3O4 nanoparticles are promising anticancer agents for targeted drug delivery.

Rhinolithiasis due to supernumerary ectopic tooth: very rare case.

An ectopic supernumerary tooth causing the formation of rhinolith was never reported before in the medical literature. A 30 years old male patient presented to our hospital with one sided nasal obstruction, recurrent epistaxis, and nasal discharge. Anterior rhinoscopy, nasal endoscopy and X-ray paranasal sinuses revealed a rhinolith in the left nasal cavity. Preoperative evaluation and post operative examination of the specimen proved that the nidus of rhinolith was a supernumerary ectopic tooth.