Potential to use cannabinoids as adjunct therapy for dexamethasone: An in vitro study with canine peripheral blood mononuclear cells.

Dexamethasone (dex) is a potent glucocorticoid used to treat a variety of diseases. It is widely used in veterinary medicine in many species; for instance, in dogs, it can be used for emergent cases of anaphylaxis or trauma, management of immune-mediated hemolytic anemia or thrombocytopenia, certain cancers, allergic reactions, and topically for skin or eye inflammation. Dex is not without its side effects, especially when administered systemically, which might compromise compliance and effective treatment. Thus, adjunct therapies have been suggested to allow for decreased dex dosing and reduction in side effects while maintaining immunosuppressive efficacy. The goal of this study was to evaluate the potential for cannabinoids to serve as adjunct therapies for dex. Immune function was assessed in canine peripheral blood mononuclear cells (PBMCs) after treatment with dex with and without cannabidiol (CBD) and/or Δ9-tetrahydrocannabinol (THC). Dex suppressed IFN-γ protein secretion in a concentration-dependent manner and this suppression by low concentrations of dex was enhanced in the presence of CBD, THC, or the combination of CBD and THC. Similar effects were found with INFG and TNFA mRNA expression. These findings provide a rationale for using CBD or THC in vivo to reduce dex dosing and side effects.

Genetic Inhibition of Mitochondrial Permeability Transition Pore Exacerbates Ryanodine Receptor 2 Dysfunction in Arrhythmic Disease.

The brief opening mode of the mitochondrial permeability transition pore (mPTP) serves as a calcium (Ca2+) release valve to prevent mitochondrial Ca2+ (mCa2+) overload. Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a stress-induced arrhythmic syndrome due to mutations in the Ca2+ release channel complex of ryanodine receptor 2 (RyR2). We hypothesize that inhibiting the mPTP opening in CPVT exacerbates the disease phenotype. By crossbreeding a CPVT model of CASQ2 knockout (KO) with a mouse missing CypD, an activator of mPTP, a double KO model (DKO) was generated. Echocardiography, cardiac histology, and live-cell imaging were employed to assess the severity of cardiac pathology. Western blot and RNAseq were performed to evaluate the contribution of various signaling pathways. Although exacerbated arrhythmias were reported, the DKO model did not exhibit pathological remodeling. Myocyte Ca2+ handling was similar to that of the CASQ2 KO mouse at a low pacing frequency. However, increased ROS production, activation of the CaMKII pathway, and hyperphosphorylation of RyR2 were detected in DKO. Transcriptome analysis identified altered gene expression profiles associated with electrical instability in DKO. Our study provides evidence that genetic inhibition of mPTP exacerbates RyR2 dysfunction in CPVT by increasing activation of the CaMKII pathway and subsequent hyperphosphorylation of RyR2.

A protocol for feasibility of plasma based GeneXpert platform and Dried Blood Spot (DBS) based Abbott platform for HIV-1 viral load testing among the people living with HIV attending ART centers in India.

BACKGROUND: HIV-1 Viral load (VL) measures efficiency of the antiretroviral therapy (ART) after treatment initiation and helps to diagnose virological failures at an early stage. Current VL assays require sophisticated laboratory facilities. As well as there are other challenges pertaining to insufficient laboratory access, cold-chain management and sample transportation. Hence the number of HIV-1 VL testing laboratories is inadequate in the resource limited settings. The revised national tuberculosis elimination programme (NTEP) in India has developed a vast network of point of care (PoC) testing facilities for diagnosis of tuberculosis and several GeneXpert platforms are functional under this programme. Both the GeneXpert HIV-1 assay and HIV-1 Abbott real time assay are comparable and GeneXpert HIV-1 assay can be used as PoC for HIV-1 Viral load testing. Also, the dried blood spot (DBS) as a sample type has been considered as a good option for HIV-1 VL testing in hard to reach areas. This protocol is therefore developed to assess the feasibility of integrating HIV-1 VL testing among people living with HIV (PLHIV) attending ART centres using the two public health models under the current programme: 1. HIV-1 VL testing using GeneXpert platform and plasma as a sample type, and 2. HIV-1 VL testing using Abbott m2000 platform and DBS as a sample type. METHODS: This ethically approved feasibility study will be implemented at two moderate to high burden ART centres where VL testing facility is not available in the town. Under Model-1, arrangements will be made to carry out VL testing on the adjacent GeneXpert facility and under Model-2, DBS will be prepared on site and couriered to identified viral load testing laboratories. In order to assess the feasibility, data will be collected on pretested questionnaire pertaining to number of samples tested for VL testing, number of samples tested for tuberculosis (TB) diagnosis and the turnaround time (TAT). In-depth interviews will be conducted among the service providers at ART centre and different laboratories for addressing any issues regarding the model implementation. RESULTS: The proportion of PLHIV tested for VL at ART centres, total TAT for both models including TAT for sample transportation, sample testing and receipt of results as well as proportion of sample rejections and reasons for the same, correlation coefficient between DBS based and plasma based VL testing will be estimated using various statistical tools. CONCLUSION: If found promising, these public health approaches will be helpful for the policy makers and program implementation in scaling up HIV-1 viral load testing within India.

Evaluation of Nanoleakage Depth and Pattern of Cervical Restorations Bonded with Different Adhesive Systems.

Aim: To evaluate nanoleakage depth and pattern of cervical restorations bonded with different adhesive systems. Materials and methods: Thirty-six extracted human premolar teeth were used for the study and grouped according to different bonding agents.Group I: fifth generation dentin bonding agent-ONE COAT SL.Group II: sixth generation dentin bonding agent-PARABOND.Group III: seventh generation dentin bonding agent-ONE COAT 7.0.For nanoleakage depth evaluation, 36 teeth were divided into three groups of 12 teeth each, according to adhesive systems used. For each adhesive system, teeth were subdivided into three subgroups of four teeth each, according to storage period, 24 hours, 1 month, and 3 months before the examination. In each tooth, two cavities were prepared (buccal and lingual), each cavity was lined with different adhesive systems and restored using a nanohybrid composite. The restored teeth were then immersed in water bath at temperature 37oC for intended period of time and then stored in 50% silver nitrate for 24 hours and photo developing solution for 8 hours. After this, the teeth were cut in buccolingual direction and subjected to scanning electron microscope (SEM) analysis for nanoleakage depth analysis. Results: Group II showed the highest nanoleakage at all three periods. At 24 hours, group III showed more leakage than group I (mean = 0.2869 > 0.2506). At 1 month storage period, there was no significant difference in the leakage. At 3 months storage period, group III showed less leakage than group I (mean = 0.5544 < 0.7313). How to cite this article: Bhupanapadu N, Sattar MA, Deb A. Evaluation of Nanoleakage Depth and Pattern of Cervical Restorations Bonded with Different Adhesive Systems. Int J Clin Pediatr Dent 2022;15(3):299-303.

SR-Mitochondria Crosstalk Shapes Ca Signalling to Impact Pathophenotype in Disease Models Marked by Dysregulated Intracellular Ca Release.

AIMS: Diastolic Ca release (DCR) from sarcoplasmic reticulum (SR) Ca release channel ryanodine receptor (RyR2) has been linked to multiple cardiac pathologies, but its exact role in shaping divergent cardiac pathologies remains unclear. We hypothesize that the SR-mitochondria interplay contributes to disease phenotypes by shaping Ca signalling. METHODS AND RESULTS: A genetic model of catecholaminergic polymorphic ventricular tachycardia (CPVT2 model of CASQ2 knockout) and a pre-diabetic cardiomyopathy model of fructose-fed mice (FFD), both marked by DCR, are employed in this study. Mitochondria Ca (mCa) is modulated by pharmacologically targeting mitochondria Ca uniporter (MCU) or permeability transition pore (mPTP), mCa uptake, and extrusion mechanisms, respectively. An MCU activator abolished Ca waves in CPVT2 but exacerbated waves in FFD cells. Mechanistically this is ascribed to mitochondria's function as a Ca buffer or source of reactive oxygen species (mtROS) to exacerbate RyR2 functionality, respectively. Enhancing mCa uptake reduced and elevated mtROS production in CPVT2 and FFD, respectively. In CPVT2, mitochondria took up more Ca in permeabilized cells, and had higher level of mCa content in intact cells vs. FFD. Conditional ablation of MCU in the CPVT2 model caused lethality and cardiac remodelling, but reduced arrhythmias in the FFD model. In parallel, CPVT2 mitochondria also employ up-regulated mPTP-mediated Ca efflux to avoid mCa overload, as seen by elevated incidence of MitoWinks (an indicator of mPTP-mediated Ca efflux) vs. FFD. Both pharmacological and genetic inhibition of mPTP promoted mtROS production and exacerbation of myocyte Ca handling in CPVT2. Further, genetic inhibition of mPTP exacerbated arrhythmias in CPVT2. CONCLUSION: In contrast to FFD, which is more susceptible to mtROS-dependent RyR2 leak, in CPVT2 mitochondria buffer SR-derived DCR to mitigate Ca-dependent pathological remodelling and rely on mPTP-mediated Ca efflux to avoid mCa overload. SR-mitochondria interplay contributes to the divergent pathologies by disparately shaping intracellular Ca signalling.

Evaluation of Immediate and Delayed Microleakage of Class V Cavities Restored with Chitosan-incorporated Composite Resins: An In Vitro Study.

AIM AND OBJECTIVE: To evaluate and compare the microleakage of unmodified microhybrid composite and 0.2% chitosan-incorporated composite in class V cavities restored immediately and after 3 months of storage in artificial saliva. MATERIALS AND METHODS: Sixty human permanent maxillary premolars were collected and standardized class V cavity prepared on the buccal surface of each tooth with dimensions: mesiodistally 3 mm, occluso cervically 2 mm, and depth of 1.5 mm and restored with microhybrid composite and chitosan-incorporated composite resins respectively and randomly divided: Group I: control-microhybrid composite (n = 30): (a) 15 teeth tested immediately (b) 15 teeth tested after 3 months. Group II-restored with chitosan + composite (n = 30): (a) 15 teeth tested immediately (b) 15 teeth tested after 3 months. Specimens were stored in artificial saliva following which a dye extraction test was carried out using a spectrophotometer. RESULTS: There was no statistically significant difference in microleakage score between the chitosan-composite group and unmodified composite group when evaluated immediately after placing the restoration. Microleakage values of the unmodified composite group increased significantly after 3 months of storage in artificial saliva and values of the chitosan-composite group did not differ significantly even after 3 months of storage. Microleakage was seen significantly less in the chitosan-composite group compared to the unmodified composite group after 3 months of storage in artificial saliva. CONCLUSION: It can be concluded that chitosan-incorporated composite seems to have improved mechanical properties and forms a more stable bond when compared with unmodified microhybrid composite in addition to being antibacterial. CLINICAL SIGNIFICANCE: Considering the advantageous properties of this material, it may be clinically useful in restoring class V cavities in patients with high caries risk. However, further in vitro and in vivo studies need to be carried out. HOW TO CITE THIS ARTICLE: Deb A, Pai V, Nadig RR. Evaluation of Immediate and Delayed Microleakage of Class V Cavities Restored with Chitosan-incorporated Composite Resins: An In Vitro Study. Int J Clin Pediatr Dent 2021;14(5):621-627.

Evaluation of Microleakage of Micro Hybrid Composite Resins versus Chitosan-Incorporated Composite Resins When Restored in Class V Cavities Using Total Etch and Self-Etch Adhesives: An In vitro Study.

AIM: To evaluate the in-vitro microleakage of traditional micro hybrid composite resin and 0.2% chitosan-incorporated composite resin when restored in Class V cavities using total etch versus self-etch adhesives after storing in artificial saliva for 24 h. MATERIALS AND METHODOLOGY: Sixty permanent maxillary premolars collected and Class V cavities were prepared on buccal surface of each tooth (dimensions: mesio-distally 3 mm, occluso cervically 2 mm, and depth of 1.5 mm) and restored with Group 1: micro hybrid (30 teeth) and Group 2: chitosan-incorporated composite (30 teeth), which was further subdivided into: (a) 15 teeth using total-etch adhesives. (b) 15 teeth using self-etch adhesives. Next dye extraction test was carried out using spectrophotometer. RESULTS: Comparison within groups: In Group 1: Self-etch demonstrated less microleakage (0.0129) compared with total etch (0.0183). The difference was statistically significant, and in Group 2: No statistically significant difference was found in mean microleakage scores after using either self-etch (0.0118) or total etch adhesives (0.0120). CONCLUSION: It can be concluded that chitosan-incorporated composite seems to have improved mechanical properties with a stable bond when used with either self-etch or total etch adhesives in addition to being antibacterial. It may be clinically useful in restoring Class V cavities in patients with high caries risk. However, further in vitro and in-vivo studies need to be carried out.