Invitro Analysis Research Articles

Development of Okra (Abelmoschus esculentus L.) Inulin extruded Snacks and Evaluation of their Prebiotic and Safety Potentials

Food industries are currently facing the challenge of meeting consumers’ demand for functional foods that provide additional health benefits. Development of appropriate extruded food snacks with inulin inclusion and its associated prebiotic benefits has not be fully exploited. This work is therefore aimed at bridging this gap. Okra pod (Abelmoschus esculentus L) inulin was blended with rice flour and other raw material inputs (sugar, salt, fat and water). These were subjected to extrusion conditions (feed rate: 300g/min, barrel temperature: 125oC and internal pressure: 20.5x10-5N/M2) for 30min. Thereafter, the extrudates were air cooled at room temperature. The resulting extrudates were subjected to bifidogenic invitro analysis using pure probiotic organisms (Bifidobacterium bifidum and Lactobacillus bulgaricus) and safety analyses using animal studies (wistar rats n=50). There were no significant differences (P<0.05) on the percentage weight gain of the experimental animals. Wistar rats fed with experimental snack samples showed progressive weight gain compared with the ones fed with control sample. Mean haemoglobin values were between 15.57mg/dl and 16.93mg/dl indicating that the snack samples are safe for human consumption. Generally, the developed snack samples were proved to have prebiotic potentials as they enhanced the growth of Bifidobacterial bifidum and Lactobacillus bulgaricus bacteria thrice more than the control sample.

In silico Repurposing of Anticancer Drug (5-Fluorouracil) as an Antibacterial Agent against Klebsiella pneumoniae

Background: Infections caused by drug-resistant microorganisms have been increasing worldwide, thereby being one of the major causes of morbidity in the 21st century. Klebsiella pneumoniae is one such bacteria causing lung inflammation, lung injury and death. The emergence of hyper-virulent and drug-resistant species, such as ESBL and CRKP, has made this microbe a serious and urgent threat. The pace of emergence of these species is outgrowing the development of novel drug and vaccine candidates, thereby shifting the research focus towards the drug repurposing approach. Objective: The study aimed at Homology Modeling of Thymidylate Synthase, verification of modeled structure, Molecular Docking and Molecular Dynamic Simulation of the docked complex, and finally, in vitro analysis of 5-FU activity against Klebsiella pneumonia. Methods: The 3D structure of Thymidylate Synthase was predicted using the Swiss model server and validated by in silico approaches. The protein-protein interactions were determined using the STRING database. Molecular Docking has been carried out and MD simulations of 5-FU with the predicted structure of thymidylate synthase have been conducted. Finally, in vitro antimicrobial drug sensitivity assay has been carried out at different concentrations. Result: Hydrogen bond was observed in Molecular Docking, protein-ligand complex remained stable during simulation, while 5-FU showed antimicrobial activity against Klebsiella pneumonia during the in vitro study. Conclusion: Both in silico as well as in vitro analysis have indicated that 5-FU can potentially be developed as an antimicrobial agent towards Klebsiella pneumonia.

Crystal structure, spectroscopic characterization, molecular docking, in vitro antibacterial potential and theoretical investigations on creatininium hydrogen maleate (CRHM) single crystal

The main focus of this work is to synthesize creatininium hydrogen maleate (CRHM) single crystal by using slow evaporation technique and to analyze its antibacterial activity by means of molecular docking and invitro analysis. Single Crystal X-ray diffraction (SXRD) analysis reveals that the grown crystal crystallizes in orthorhombic structure with non centrosymmetric space group P212121 and the optimized structural parameters explains the structural stability with N-H...O, C-H…O and O-H…O interactions. Hirshfeld surface analysis of grownup single crystal was performed to check and ensure the presence of intermolecular interactions that creates the molecule more stable. Natural Bond Orbital (NBO) analysis provides an efficient method for studying intermolecular and intramolecular hydrogen bonding, intermolecular charge transfer and delocalization of electron density. FTIR and Raman spectroscopy reveals the functional group associated with grown crystal along with absorption peaks and bands were studied and assigned in theoretical and experimental modes. The computed and experimental value of 1H and 13C NMR chemical shifts were performed. By TG/DTA analysis, it has been analyzed that the grown crystal is thermally stabile up to 165 °C. The mechanical strength of the grown crystal and work hardening coefficient is found to be 3 by using Vicker's microhardness test and confirms the soft nature that favors the bioactivity. The electronic transition of the materials were analyzed from UV- Visible absorbence spectrum it indicates that they exhibits good optical transparency in the UV–Visible and near IR region having lower cut-off wavelength of 310 nm and optical band gap of 3.9 eV. The green fluorescence nature of the synthesized material was analyzed by fluorescence spectral analysis. Frontier molecular orbital analysis has been carried out to explain the charge transfer within the molecule that enhances the antibacterial nature of the molecule and energy gap of the material was found to be 5.11 eV. Molecular electrostatic surface analysis revealed that positive potential is confined around the creatinine portion and negative region is scattered over the maleate portion exhibits an excellent ICT interactions exists within the lattice. The antibacterial activities of the compound have been screened by molecular docking. Moreover, the prepared compound is screened for its in vitro antibacterial activity against various types of bacteria, Escherichia coli, Pseudomonas aeruginosa, Serraitia marcescens, Bacillus cereus and Staphylococcus aureus. The results of these studies indicated that the title compound exhibit excellent antibacterial efficiency.

In-Silico and In-Vitro Analysis of Human SOS1 Protein Causing Noonan Syndrome - A Novel Approach to Explore the Molecular Pathways.

AimsPerform in-silico analysis of human SOS1 mutations to elucidate their pathogenic role in Noonan syndrome (NS).BackgroundNS is an autosomal dominant genetic disorder caused by single nucleotide mutation in PTPN11, SOS1, RAF1, and KRAS genes. NS is thought to affect approximately 1 in 1000. NS patients suffer different pathogenic effects depending on the mutations they carry. Analysis of the mutations would be a promising predictor in identifying the pathogenic effect of NS.MethodsWe performed computational analysis of the SOS1 gene to identify the pathogenic nonsynonymous single nucleotide polymorphisms (nsSNPs) th a t cause NS. SOS1 variants were retrieved from the SNP database (dbSNP) and analyzed by in-silico tools I-Mutant, iPTREESTAB, and MutPred to elucidate their structural and functional characteristics.ResultsWe found that 11 nsSNPs of SOS1 that were linked to NS. 3D modeling of the wild-type and the 11 nsSNPs of SOS1 showed that SOS1 interacts with cardiac proteins GATA4, TNNT2, and ACTN2. We also found that GRB2 and HRAS act as intermediate molecules between SOS1 and cardiac proteins. Our in-silico analysis findings were further validated using induced cardiomyocytes (iCMCs) derived from NS patients carrying SOS1 gene variant c.1654A>G (NSiCMCs) and compared to control human skin fibroblast-derived iCMCs (C-iCMCs). Our in vitro data confirmed that the SOS1, GRB2 and HRAS gene expressions as well as the activated ERK protein, were significantly decreased in NS-iCMCs when compared to C-iCMCs.ConclusionThis is the first in-silico and in vitro study demonstrating that 11 nsSNPs of SOS1 play deleterious pathogenic roles in causing NS.

Dietary Preference does not affect Salivary Total Antioxidant Capacity of Group of young and Middle-aged Adults in South India – An In-Vitro Analysis

Background: Oxidative stress is a pathological state resulting from excess free radical activity in the body. Studies show associations between oxidative stress and cancer. The saliva has anti-oxidant mechanisms that may play a role in preventing/fighting oral cancer. There is public health interest in determining dietary influences on salivary antioxidant capacity. Materials and method: Age and sex-matched participants were selected based on eligibility criteria after informed consent. A diet questionnaire and chart was administered to determine dietary preferences. They were then divided into two groups – vegetarian and non-vegetarian. Total salivary antioxidant capacity of unstimulated saliva samples was analyzed using spectrophotometry and compared between the two groups. Results and inference: 30 participants selected were categorized in two groups included in the study. Statistical analysis of the spectrophotometric findings indicated that there were no significant differences between the groups. Conclusion: Total salivary antioxidant capacity did not significantly differ between vegetarians and non-vegetarians. Therefore, the implication may be that, either the type of diet may not influence antioxidant capacity of saliva, or the effect of diet may not reflect in the salivary antioxidant capacity.

Leveraging health systems data to characterize a large effect variant conferring risk for liver disease in Puerto Ricans

The integration of genomic data into health systems offers opportunities to identify genomic factors underlying the continuum of rare and common disease. We applied a population-scale haplotype association approach based on identity-by-descent (IBD) in a large multi-ethnic biobank to a spectrum of disease outcomes derived from electronic health records (EHRs) and uncovered a risk locus for liver disease. We used genome sequencing and in silico approaches to fine-map the signal to a non-coding variant (c.2784-12T>C) in the gene ABCB4. Invitro analysis confirmed the variant disrupted splicing of the ABCB4 pre-mRNA. Four of five homozygotes had evidence of advanced liver disease, and there was a significant association with liver disease among heterozygotes, suggesting the variant is linked to increased risk of liver disease in an allele dose-dependent manner. Population-level screening revealed the variant to be at a carrier rate of 1.95% in Puerto Rican individuals, likely as the result of a Puerto Rican founder effect. This work demonstrates that integrating EHR and genomic data at a population scale can facilitate strategies for understanding the continuum of genomic risk for common diseases, particularly in populations underrepresented in genomic medicine.

In-vitro evaluation of electrospun cellulose acetate nanofiber containing Graphene oxide/TiO2/Curcumin for wound healing application

Graphene-based dressings have gained interest in wound healing due to their biocompatibility and antimicrobial properties. This study developed a graphene oxide/TiO 2 /curcumin integrated cellulose acetate (CA) nanofiber for wound dressing applications. Electrospinning was used to fabricate the CA/GO/TiO 2 /curcumin nanofiber. The formation of CA nanofiber was clearly visible using FE-SEM. The presence of curcumin in CA/GO/TiO 2 /curcumin nanofiber was confirmed by FT-IR analysis. The prepared nanofiber was characterized by swelling, degradation, tensile strength, and in-vitro wound healing properties. CA/GO/TiO 2 /curcumin nanofiber demonstrated considerable antibacterial activity against wound pathogens and improved wound healing, as confirmed by MTT and migration assays. The fabricated CA/GO/TiO 2 /curcumin nanofiber exhibited exceptional hemocompatibility and biocompatibility. The drug release study revealed that sustained curcumin release would improve skin regeneration and wound healing. Invitro analysis confirmed that CA/GO/TiO 2 /curcumin nanofiber has the potential to be used as a wound dressing material. • Graphene oxide/TiO 2 /curcumin incorporated cellulose acetate (CA) nanofiber for wound dressing applications. • The CA/GO/TiO 2 /curcumin nanofiber was fabricated by electrospinning method. • Composite nanofibers enhances wound healing process which is confirmed by MTT and migration assay. • The sustained drug release behaviour would enhance skin regeneration. • In vitro analysis confirms that the CA/GO/TiO 2 /curcumin nanofiber is a suitable wound dressing material.

HADHB, a fatty acid beta-oxidation enzyme, is a potential prognostic predictor in malignant lymphoma

In haematological malignancies, such as malignant lymphoma, reprogramming of fatty acid metabolism favours tumour cell survival and drug resistance. Hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha (HADHA), an enzyme involved in fatty acid beta-oxidation (FAO), is overexpressed in high-grade lymphoma and is a predictor of poor prognosis in diffuse large B-cell lymphoma (DLBCL). HADHB forms a heterodimer with HADHA and functions as an FAO enzyme together with HADHA; however, the relevance of its expression in malignant lymphoma is unknown. In this study, we investigated the roles and antitumour effects of HADHB expression in malignant lymphoma. Immunohistochemical analysis showed that HADHB was frequently overexpressed in the high-grade lymphoma subtype. HADHB overexpression was observed in 68% (87/128) of DLBCL cases and was an independent predictor of poor prognosis (p=0.001). Invitro analysis demonstrated that HADHB knockdown suppressed cell proliferation in LCL-K and MD901 cells (p<0.05). Additionally, treatment with the FAO inhibitor, ranolazine, increased cell death in control cells compared with that in HADHB knockdown LCL-K and MD901 cells (p<0.01). Cell death was also suppressed by the ferroptosis inhibitor, ferrosatin-1, in LCL-K and MD901 cells (p<0.05). Collectively, these findings provide basic evidence for the development of new cell death-based therapies for refractory malignant lymphoma. We plan to perform prospective studies and preclinical studies using animal models to confirm these results.

Anti-COVID-19 activity of some benzofused 1,2,3-triazolesulfonamide hybrids using in silico and in vitro analyses

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a pandemic fatal infection with no known treatment. The severity of the disease and the fast viral mutations forced the scientific community to search for potential solution. Here in the present manuscript, some benzofused1,2,3triazolesulfonamide hybrids were synthesized and evaluated for their anti- SARS-CoV-2 activity using in silico prediction then the most potent compounds were assessed using in-Vitro analysis. The in-Silico study was assessed against RNA dependent RNA polymerase, Spike protein S1, Main protease (3CLpro) and 2′-O-methyltransferase (nsp16). It was found that 4b and 4c showed high binding scores against RNA dependent RNA polymerase reached −8.40 and −8.75 ​kcal/mol, respectively compared to the approved antiviral (remdesivir −6.77 ​kcal/mol). Upon testing the binding score with SARS-CoV-2 Spike protein it was revealed that 4c exhibited the highest score (−7.22 ​kcal/mol) compared to the reference antibacterial drug Ceftazidime (−6.36 ​kcal/mol). Surprisingly, the two compounds 4b and 4c showed the highest binding scores against SARS-CoV-2 3CLpro (−8.75, −8.48 ​kcal/mol, respectively) and nsp16 (- 8.84 and – 8.89 ​kcal/mol, respectively) displaying many types of interaction with all the enzymes binding sites. The derivatives 4b and 4c were examined in vitro for their potential anti-SARS-CoV-2 and it was revealed that 4c was the most promising compound with IC50 reached 758.8108 ​mM and complete (100%) inhibition of the binding of SARS-CoV-2 virus to human ACE2 can be accomplished by using 0.01 ​mg.

THE ROLE OF INTERMEDIATE IRRIGANTS IN THE PREVENTION OF THE FORMATION OF ORANGE-BROWN PRECIPITATE, FORMED DUE TO INTERACTION BETWEEN SODIUM HYPOCHLORITE AND CHLORHEXIDINE: A SPECTROPHOTOMETRIC ANALYSIS.

This study aimed to assess the efciency of intermediate irrigants in the prevention of formation of precipitate, formed due to the interaction of sodium hypochlorite and chlorhexidine when used as root canal irrigants.The formation of orange brown precipitate, due to the reaction of 3% sodium hypochlorite and 2% chlorhexidine was assessed using spectrophotometric analysis. Three intermediate irrigants were then assessed for their capabilities to prevent the formation of the precipitate. Group A- 6.25% sodium metabisulphite, Group B- 10% sodium ascorbate and Group C- saline were used at different volumes and the values were recorded by measuring their optical density. An invitro analysis was further carried out on single rooted teeth, to verify the efcacy of intermediate irrigants. The teeth which were subjected to the tests were sectioned and observed under stereomicroscope for the presence of remnants of orange brown precipitate in the root canals. Results: The spectrophotometric analysis showed peak value at 450nm and the precipitate formation was found at 60µl of 2% chlorhexidine and 1 ml of sodium hypochlorite. There was statistical difference in the optical density values between the groups, with saline having highest optical density values followed by 6.25% sodium metabisulphite and 10% sodium ascorbate group. Wherein 10% sodium ascorbate solution prevented the formation of orange brown precipitate than other two group. It was concluded that 10% sodium ascorbate played a vital role in the reduction of formation of parachloro-aniline, when used as an intermediate irrigant. 6.25% Sodium metabisulphite also prevented the precipitate formation, to a lesser extent than sodium ascorbate. However, saline did not eliminate the formation of the precipitate.

A liquid culture cancer spheroid model reveals low PI3K/Akt pathway activity and low adhesiveness to the extracellular matrix.

Three-dimensional (3D) cultures of cancer cells in liquid without extracellular matrix (ECM) offer invitro models for metastasising conditions such as those in vessels and effusion. However, liquid culturing is often hindered by cell adhesiveness, which causes large cell clumps. We previously described a liquid culture material, LA717, which prevents nonclonal cell adhesion and subsequent clumping, thus allowing clonal growth of spheroids in an anchorage-independent manner. Here, we examined such liquid culture cancer spheroids for the acquisition of apical-basal polarity, sensitivity to an Akt inhibitor (anticancer drug MK-2206) and interaction with ECM. The spheroids present apical plasma membrane on the surface, which originated from the failure of polarisation at the single-cell stage and subsequent defects in phosphorylated ezrin accumulation at the cell boundary during the first cleavage, failing internal lumen formation. At the multicellular stage, liquid culture spheroids presented bleb-like protrusion on the surface, which was enhanced by the activation of the PI3K/Akt pathway and reduced by PI3K/Akt inhibitors. Liquid culture spheroids exhibited slow proliferation speed and low endogenous pAkt levels compared with gel-cultured spheroids and 2D-cultured cells, explaining the susceptibility to the Akt-inhibiting anticancer drug. Subcutaneous xenografting and invitro analysis demonstrated low viability and adhesive property of liquid culture spheroids to ECM, while migratory and invasive capacities were comparable with gel-cultured spheroids. These features agree with the low efficacy of circulating tumour spheroids in the settling step of metastasis. This study demonstrates the feature of anchorage-independent spheroids and validates liquid cultures as a useful method in cancer spheroid research.

Preparation of biocompatible polymeric bio-scaffold as wound healing activity: Characterization and In vivo analysis

Abstract In this study, a combination of collagen (CL), gelatin (GL) and squid ink (SI) were used to produce a biocompatible polymeric bio-scaffold (CL, CL: GL and CL: GL: SI) was examined for wound healing purposes. Biocompatible polymeric bio-scaffold (BPBS) plays an important role in wound care due to their cost-effective, good mechanical properties, bio-compatible and antimicrobial characteristics. Thermogravimetric analysis (TGA) and Fourier transform infrared spectroscopy (FTIR) of the BPBS has showed the heat stability and nature of samples, respectively, and while scanning electron microscopy (SEM) images revealed the blending of CL fiber, GL, and SI pattern interface. BPBS displayed excellent antimicrobial activity against both E.coli and S.aureus. The inhibition zone of BPBS results obtained in 4.57 ± 0.30 mm against E.coli and 4.74 ± 0.15 mm against S.aureus. Mechanical characteristics of BPBS shows tensile strength of 20.88 ± 0.05 (Mpa), 17.74 ± 0.05 (%) as Elongation at break, 26.31 ± 0.21 (%) of flexibility and water absorption of 29.80 ± 0.05 (%). Invitro analysis of CL: GL: SI, displayed 100% biocompatibility and cell viability was found 3rd days. In vivo study using mice by excisional animal model examined the CL: GL: SI scaffold can significantly accelerate the process of wound healing.

The Effect of Biopolymer Coatings on Hernia Meshes in a Rat Model

Purpose: To improve biocompatibility and texture of hernia meshes has played a key role in tissue engineering for decades. Biopolymer (Polyethylenimine (PEI) and 3-Glycidoxypropyltrimethoxysilane (GOPS)) coating on Polypropylene (PP) and expanded Polytetrafluoroethylene (ePTFE) mesh showed promising results in fibroblast adhesion and cell growth in an invitro analysis. The objective of this animal study was to evaluate whether this may influence the incorporation into host tissue. Methods: 30 male Lewis rats were divided into 3 groups (n=10): Group 1: ePTFE/PEI, Group 2: ePTFE/GOPS, Group 3: PP/PEI. In each animal, a 3x0.5 cm coated mesh was implanted in the right rectus sheath in sublay position, the uncoated mesh was implanted on the left equally. After 90 days, the rats were sacrificed and each side of the rectus sheath was analyzed separately for adhesions and mechanical strength. Histopathological assessment included Gieson’s stain and haematoxylin-eosin staining. The Wilcoxon test was used for statistical analyses. Results: The GOPS-coated ePTFE tends to cause more adhesions. There was no significant difference in the mechanical strength within and between the groups, but the PEI-coated polypropylene was significantly less extendible (p&lt;0.05) compared to the uncoated PP. In group 2, Gieson’s stain showed a significantly lower surrounding tissue reaction of foreign-body giant cells and scar tissue around the PEI-coated mesh compared to the uncoated ePTFE (p&lt;0.05). Conclusions: It is possible to coat surgical mesh devices with biopolymers. They do not lead to a lack of mechanical strength. The GOPS-coating did not show any general positive effect on the biocompatibility of meshes. The PEIcoating resulted in a lower surrounding tissue reaction and in a less extendible abdominal wall and should therefore be investigated further.

Effects of temperature on the nitrate reductase activity and growth of Ulva prolifera.

To better understand the effect of temperature on the growth and nitrate reductase activity (NRA) of Ulva prolifera and their relationships, the effects of five different temperatures (10, 15, 20, 25, and 30°C) were investigated in a laboratory setup. In this study, an optimization invitro analysis method for Ulva prolifera NRA was developed. Under different treatments, the NRA, nitrate concentration, pH, the intracellular nitrate and nitrite concentrations, and the POC/PON were evaluated. The results of the invitro analysis method showed it was optimal for the NRA assay when the extraction time was 6min, enzymatic reaction time 30min, volume of phenazine methosulfate (PMS) solution 50μL, NADH concentration 0.36mM, and KNO3 concentration 10mM. The maximal NRA (NRAmax ) appeared on the 2nd day in the 10, 15, and 20°C (low-temperature) groups and on the 1st day in the 25 and 30°C (high-temperature) groups. The algal growth ended earlier at a high temperature, ending after 5d at 30 and 25°C and 7d at 20°C and 9d at 15°C, and the alga at 10°C had been growing during the incubation period. Ulva prolifera cultivated in a range of 10-20°C had a long growth cycle and the NRA decreased with increasing temperature when exceeded 15°C, a positivecorrelation between algal growth and NRA was observed. This study supports NRA is a suitable proxy of the effects of temperature changes on the ability of Ulva prolifera to uptake and metabolize nitrogen nutrients.

Particle size, penetration rate and effects of smoke and smokeless tobacco products – an invitro analysis

Utilization of tobacco products is a global epidemic that causes a health threat among the adolescents and adults of the world. If the current pattern perpetuates, it is estimated to cause 10 million deaths each year by 2020. According to the 2009–2010 survey by Global Adult Tobacco Survey, 53.5% of Indians use tobacco products (66.2% of men and 40% of women). Cancers of the lip and oral cavity are highly frequent in Southern Asia (eg: India and Sri Lanka) and it is also the leading cause of cancer death among men in India and Sri Lanka. Aim and objectivesThe aim of the study is to evaluate the particle size, and assess the penetration rate in egg shell membrane and proliferation rate using cell lines among 4 commercially available tobacco products. Materials and methodsA total of 4 tobacco product samples namely Gutka, Khaini, Beedi and Cigarette was used to analyse the particle size using High resolution Scanning electron microscopy, the penetration rate on egg shell membrane using Spectroflurometry and proliferative rate using ELISA on cell lines respectively. ResultsKhaini revealed the smallest particle size and highest penetration and proliferation rate compared to Gutka, Beedi and Cigarette. ConclusionCurrent evidence shows that individuals who consume Khaini are at high risk to develop potentially malignant disorders and oral squamous cell carcinoma when compared to Gutka, Beedi and Cigarette (in the order of occurrence).

Detailed analysis of Japanese patients with adenosine deaminase 2 deficiency reveals characteristic elevation of type II interferon signature and STAT1 hyperactivation

Deficiency of adenosine deaminase 2 (DADA2) is an autosomal recessive inflammatory disease caused by loss-of-function mutations in both alleles of the ADA2 gene. Most patients with DADA2 exhibit systemic vasculopathy consistent with polyarteritis nodosa, but large phenotypic variability has been reported, and the pathogenesis of DADA2 remains unclear. This study sought to assess the clinical and genetic characteristics of Japanese patients with DADA2 and to gain insight into the pathogenesis of DADA2 by multi-omics analysis. Clinical and genetic data were collected from 8 Japanese patients with DADA2 diagnosed between 2016 and 2019. ADA2 variants in this cohort were functionally analyzed by invitro overexpression analysis. PBMCs from 4 patients with DADA2 were subjected to transcriptome and proteome analyses. Patient samples were collected before and after introduction of anti- TNF-α therapies. Transcriptome data were compared with those of normal controls and patients with other autoinflammatory diseases. Five novel ADA2 variants were identified in these 8 patients and were confirmed pathogenic by invitro analysis. Anti-TNF-α therapy controlled inflammation in all 8 patients. Transcriptome and proteome analyses showed that upregulation of type II interferon signaling was characteristic of DADA2. Network analysis identified STAT1 as a key regulator and a hub molecule in DADA2 pathogenesis, a finding supported by the hyperactivation of STAT1 in patients' monocytes and B cells after IFN-γ stimulation. Type II interferon signaling and STAT1 are associated with the pathogenesis of DADA2.

An In-Vitro Analysis of Peri-Implant Mucosal Seal Following Photofunctionalization of Zirconia Abutment Materials

The presence of epithelial and connective tissue attachment at the peri-implant–soft tissue region has been demonstrated to provide a biological barrier of the alveolar bone from the oral environment. This barrier can be improved via surface modification of implant abutment materials. The effect of photofunctionalization on creating a bioactive surface for the enhancement of the epithelial and connective tissue attachment of zirconia implant abutment’s peri-implant mucosal interface using organotypic model has not been investigated. Therefore, this study aimed to evaluate the soft tissue seal around peri-implant mucosa and to understand the effect of photofunctionalization on the abutment materials. Three types of abutment materials were used in this study; yttria-stabilized zirconia (YSZ), alumina-toughened zirconia, and grade 2 commercially pure titanium (CPTi) which were divided into nontreated (N-Tx) and photofunctionalized group (UV-Tx). The three-dimensional peri-implant mucosal model was constructed using primary human gingival keratinocytes and fibroblasts co-cultured on the acellular dermal membrane. The biological seal was determined through the concentration of tritiated water permeating the material–soft tissue interface. The biological seal formed by the soft tissue in the N-Tx group was significantly reduced compared to the UV-treated group (p < 0.001), with YSZ exhibiting the lowest permeability among all materials. Photofunctionalization of implant abutment materials improved the biological seal of the surrounding soft tissue peri-implant interface.

Comparative In-Vitro Analysis on Anti-Diabetic Potential of Selected Medicinal Herbs of Nepal

Comparative In-Vitro Analysis on Anti-Diabetic Potential of Selected Medicinal Herbs of Nepal

In-Vitro analysis of medicinal plant for anti-inflammatory properties

This examination demonstrations the in vitro mitigating impact of leaf concentrates of Polyalthia longifolia. These were investigated in various measures to decide their mitigating impacts. Protein denaturation and RBC film adjustment were measured. The outcomes were accounted for as % hindrance in triplicate assurance and were exposed to factual additionally, diclofenac Aspirin and sodium were utilized as norms. The consequence of the % restraint of protein denaturation demonstrated that at 1mg/ml diclofenac, ethanolic extricate, new leaf fluid concentrate and dry leaf watery concentrate. Because of dissecting the MS and UV information and in contrast with the legitimate norms, an aggregate of 10 polyphenols were recognized and measured, counting gallic corrosive, catechin, quercetin, kaempferol, and their subordinates. This is the primary investigation looking at the leaves, as opposed to the organic product or seeds reasonably collected yet underutilized by nearby indigenous people groups. The concoction outline of the leaf was thoroughly broke down and prompted the ID of 10 polyphenol mixes, counting phenolic corrosive and flavonoids. The individual polyphenols were effectively quantitated, utilizing UV recognition. Additional bioactivity examinations demonstrated that the concentrates of leaf display cancer prevention agent, antiproliferation, and calming exercises. The primary atomic component may mostly be donated by the hindrance of NF-κB enactment, a typical sign alleyway among multiplication and aggravation. Additional examinations are expected to investigate whether these polyphenol mixes could work synergistically to accomplish significantly better exercises than each solitary segment. The concoction bioactivities and profile decided to help its conventional usage and might assistance for its additional pharmacological investigations and nutraceutical requests.

Investigating the Cytotoxic and Anti-Bacterial Activity of Commercially Available Local Anesthetics: An In-Vitro Analysis

OBJECTIVE: To evaluate and compare the influence of three local anesthetic dental formulations manufactured in France (Septodont), Korea (Medicaine) and Pakistan (HD-Caine) in terms of cytotoxicity and anti-bacterial activities. METHODOLOGY: 90 commercially available local anesthetic cartridges of similar composition (2% lidocaine with epinephrine 100,000) viz. Septodont, Medicaine and HD-Caine were randomly collected from three different Pakistani cities and were assigned as Group S, Group M and Group H, respectively. The cartridges were further divided into three sub-groups each consisting of 10 cartridges to first evaluate cytotoxicity on Mesenchymal Stem Cells (MSCs) using a flow cytometer and secondly to investigate anti-bacterial activity by measuring zones of inhibition and through Broth Dilution Method against five bacterial strains. RESULTS: The results indicated that Septodont (94.5±0.1) and Medicaine (94.7±0.0) showed the highest viability percentage with no significant difference when the two were compared (P=0.6). HD-Caine (93.9±0.0) showed the least, being significantly (P&lt;0.01) different from Septodont and Medicane. A statistically significant (P&lt;0.05) difference was identified between the three study groups regarding the anti-bacterial activity. HD-Caine showed the highest anti-bacterial potential, followed by Medicaine and Septodont. CONCLUSION: Mild toxicity was observed by all the three groups in human MSCs, justifying their safe use in clinical practice. Additionally, Medicane and HD-Caine showed significant anti-bacterial activity indicating their possible use as sterile irrigants. KEYWORDS: Dental anesthesia, Lidocaine, Epinephrine, Antibacterial activity HOW TO CITE: Imran E, Moeen F, Satti H, Rahman L. Investigating the cytotoxic and anti-bacterial activity of commercially available local anesthetics: An in-vitro analysis. J Pak Dent Assoc 2020;29(4):185-192.