Anti-insomnia Effect of a Polyherbal Formulation on P-chlorophenyalanine Induced Experimental Animal Model.

Sleep is a dynamic and controlled set of physiological and behavioural practices during which the stabilisation and restoration processes of the body take place properly. Therefore, sleep disorders, especially chronic insomnia, can harm an individual's physical and mental health. However, the therapeutic alternatives are limited and possess severe side effects. Thus, in this study, we aimed to investigate the anti-insomnia effect of a polyherbal formulation (Sleep) (SLP) on p-chlorophenyalanine (PCPA) induced insomnia in rats. Intraperitoneal injection of PCPA induced the experimental condition, and the therapeutic effect of SLP was evaluated by studying the sleep pattern and expression of various neurotransmitters and receptors, along with neurotrophins. Moreover, insomnia-associated oxidative stress and inflammation were also studied. From the findings, we found that the SLP-supplemented animals improved their sleeping behaviour and that the major neurotransmitters, hormones, and receptors were maintained at an equilibrium level. Furthermore, the neurotrophin level was increased and pro-inflammatory cytokines were reduced. The evaluation of oxidative stress markers shows that the antioxidants were significantly boosted, and as a result, lipid peroxidation was prevented. The overall findings suggest that SLP can be used as an effective medication for the treatment of sleep disorders like insomnia as it triggers the major neurotransmitter system.

Ameliorating effect of polyherbal formulation (Febrojith) on acute and chronic inflammatory model via inhibiting inflammation and oxidative stress.

ETHNOPHARMACOLOGICAL RELEVANCE: Inflammation is a complex biological response of the tissue to noxious stimuli, which causes several debilitating inflammatory disorders. Currently, various conventional medicines are available, but their consumption causes adverse effects, hence researchers focused on alternatives like medical herbs from natural sources, as one of the most promising sources of therapeutic agents for inflammation. Febrojith is a well-known traditional Ayurvedic formulation obtained from the treasures of Ayurveda with a unique blend of herbs that are used effectively in preventing and combating a broad spectrum of infections, fevers, and also enhancing immunity for many years. However, its anti-inflammatory, efficacy and underlying mechanism remained unexplored. AIM OF THE STUDY: In the present study, we investigated the chemical characterization and in vivo anti-inflammatory efficacy of Febrojith (FB) on acute and chronic inflammatory models via inhibiting inflammation and oxidative stress. MATERIALS AND METHODS: FB was analyzed for chemical characterization & its phytoconstituents by UV-Vis spectrum, FT-IR, and GC-MS analysis. The anti-inflammatory activity of FB was studied on carrageenan-induced acute and adjuvant-induced chronic experimental models. The inflammatory cytokines and mediators were measured using the ELISA & Colorimetry techniques. Histopathology and cytology of paw tissue and synovium were analyzed by H&E and Papanicolau's (PAP)-staining methods. RESULTS: 100 mg/kg bwt was found to be a potent dose from the carrageenan model and evaluated its effect in the adjuvant-induced chronic arthritic model. In the chronic model, 84% of edema inhibition was observed at the dose of 100 mg/kg bwt. Moreover, the supplementation of FB was shown to significantly (p ≤ 0.05) decrease the TBARS level and activity of myeloperoxidase in the paw tissue. In addition, adjuvant-induced production of various pro-inflammatory cytokines like TNF-α, IL-1ß, IL-6, PGE2, NO and COX-2 were suppressed in inflamed rats subjected to FB supplementation. It also revealed that FB supplementation significantly (p ≤ 0.05) reduced the haematological markers. From the histopathology and cytological analysis, we found a reduction in the edema formation, and infiltration of inflammatory cells after the supplementation of FB. CONCLUSION: In conclusion, FB might be used as an effective and potent drug against inflammation.

Recent breakthroughs in nanostructured antiviral coating and filtration materials: a brief review.

COVID-19 persists as the most challenging pandemic of the 21st century with a high rate of transmission. The main pathway of SARS-CoV-2 transmission is aerosol-mediated infection transfer through virus-laden droplets that are expelled by infected people, whereas indirect transmission occurs when contact is made with a contaminated surface. This mini review delivers an overview of the current state of knowledge, research directions, and applications by examining the most recent developments in antiviral surface coatings and filters and analyzing their efficiencies. Reusable masks and other personal protective devices with antiviral properties and self-decontamination could be valuable tools in the fight against viral spread. Moreover, antiviral surface coatings that repel pathogens by preventing adhesion or neutralize pathogens with self-sanitizing ability are assumed to be the most desirable for terminating indirect transmission of viruses. Although many nanomaterials have shown high antiviral capacities, additional research is unquestionably required to develop next-generation antiviral agents with unique characteristics to face future viral outbreaks.

Association of Mothers' Genetic Taste Perception to Eating Habits and Its Influence on Early Childhood Caries in Preschool Children: An Analytical Study.

Aims: To examine (1) the association of mothers' 6-n-propylthiouracil (PROP) sensitivity with snacking habits and caries experience of both the mother and their 2-5-year-old preschool children and (2) the association of feeding habits with caries prevalence in 2-5-year-old children. Materials and methods: In this analytical cross-sectional study, 2-5-year-old preschool children were clinically examined by a single calibrated examiner and divided into two groups of 80 children with early childhood caries (ECC) and 80 without. Mothers of the children were clinically examined and a second blinded investigator determined their genetic taste type using PROP. Data regarding feeding practices of the child and snacking habits of the mother and child were obtained using questionnaires. The data obtained were statistically analyzed. Results: Cariogenic snack consumption was more among nontaster mothers and their children (p value < 0.001). More number of children of the nontaster mothers in the ECC group were bottle-fed with sweetened beverages (n = 24) and received sweet snacks as rewards (n = 32). Caries prevalence of 73%, 20%, and 7% was seen in children of nontaster, medium taster, and supertaster mothers, respectively (p value < 0.05). Significant association was found between mother's caries activity, past bottle-feeding, receiving sweet snacks as reward and their child's caries experience. Conclusion: Mothers' PROP sensitivity was significantly associated with snacking habits and caries experience of their children. Mothers' PROP type could be an important factor that may determine and explain the dental caries experience in their 2-5-year-old preschool children. Clinical significance: Early identification of nontaster mothers may prove beneficial in guiding them and consecutively their children toward healthy dietary habits, leading to lesser incidence of caries and their better overall wellbeing. How to cite this article: Nellamakkada K, Patil SS, Kakanur M, et al. Association of Mothers' Genetic Taste Perception to Eating Habits and Its Influence on Early Childhood Caries in Preschool Children: An Analytical Study. Int J Clin Pediatr Dent 2022;15(S-2):S135-S139.

Anti-inflammatory and anti-COVID-19 effect of a novel polyherbal formulation (Imusil) via modulating oxidative stress, inflammatory mediators and cytokine storm.

In the current scenario, most countries are affected by COVID-19, a pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that has a massive impact on human health. Previous studies showed that some traditionally used medicinal herbs and their combinations showed synergistic anti-viral and anti-inflammatory activity against SARS-CoV-2 type infections. Therefore, the goal of this study is to demonstrate the anti-viral and anti-inflammatory effects of a novel polyherbal formulation, hereinafter referred to as Imusil, on Vero E6 cell lines and Raw 264.7 murine macrophage cells respectively. The Imusil was subjected to identify its chemical characterisations such as UV-Visible spectrum profile, Fourier transform infrared spectroscopy (FT-IR) and gas chromatography-mass spectroscopic (GC-MS) analysis. FT-IR analysis of Imusil peak values with various functional compounds such as alcohol, esters, aliphatic and carboxylic acids. GC-MS analysis of compounds with totally 87 compounds major chemical compounds were identified, such as 3-(Octanoyloxy) propane-1,2-diyl bis(decanoate), Succinic acid, 2-methylhex-3-yl 2,2,2-trifluoroethyl ester, Neophytadiene, 3,5,9-Trioxa-4-phosphaheneicosan-1-aminium, 4-hydroxy-N,N,N-trimethyl-10-oxo-7-[(1-oxododecyl)oxy]-, hydroxide, inner salt, 4-oxide, (R)-. The anti-viral activity of Imusil against SARS-CoV-2 was assessed using plaque reduction assay and anti-inflammatory study was conducted on lipopolysaccharide (LPS)-induced RAW 264.7 cells. The results obtained from the study reveal that Imusil significantly inhibited SARS-CoV-2 replication in Vero E6 cells and the production of inflammatory mediator's cyclooxygenase-2 and pro-inflammatory cytokines like tumour necrosis factor-α and interleukin- 6 were significantly reduced, along with thwarting the significant oxidative stress by preventing the expression of NOX-2 thereby inhibiting the reactive oxygen species formation. Hence, considering the current study as a novel strategy for mediating the COVID-19 associated aliments, inceptive scientific evidence of Imusil promises its potential therapeutic implications against COVID-19 and inflammatory conditions.

A cross-sectional cariogram-based comparison of caries risk profile in children with various levels of intellectual disability.

AIM: To compare the caries risk profile in children with mild, moderate, and severe intellectual disability (ID) using cariogram. MATERIALS AND METHODS: A cross-sectional study was conducted to compare caries risk profile using cariogram among 150 children aged 6-18 years, with various levels of ID (50 each in Mild ID, Moderate ID, and Severe ID groups), attending special schools in Bengaluru city. The risk assessment consisted of: a questionnaire, an interview, estimation of oral hygiene, saliva sampling, microbial evaluation and clinical examination. Statistical analysis of the results obtained was performed using Chi-square/Fischer's exact test, Kruskal-Wallis test and Mann-Whitney test. RESULTS: The overall comparison showed significant differences between the groups across all parameters except diet (P = 0.131), fluoride exposure (P = 1), salivary flow rate (P = 0.3), and buffer capacity (P = 1). Caries risk was high among children with severe intellectually disability while the actual chance to avoid caries was found to be high in the Mild ID group. Bacteria was one of the dominant caries risk sectors followed by susceptibility, circumstances, and diet. CONCLUSION: Cariogram can be used as an effective tool in assessing caries prediction, thus aiding in identifying different risk groups in a community so that appropriate preventive measures can be implemented to overcome the caries risk.

A clinical evaluation of two electronic apex locators and conventional radiography in working length determination in primary molar and its influence on children's behavioral responses.

BACKGROUND: Accurate root canal length determination, simplicity of the procedure, and patient's cooperation are important determinants in the success of endodontic treatment in primary teeth. AIM: The aim of the study was to clinically compare the accuracy of Propex Pixi and Formatron D10 apex locators with conventional radiography in determining working length in mandibular primary second molars with and without varying degrees of physiological root resorption. DESIGN: A nonrandomized clinical trial was conducted in 115 mandibular primary second molars indicated for pulpectomy in 90 pediatric dental patients. A single calibrated examiner determined the working length by conventional radiographic method and electronic apex locator (EAL) methods in a total of 376 canals. Behavior of the children during execution of each method in each tooth was recorded using the Frankl's behavior rating scale. The data obtained were tabulated and subjected to statistical analysis. RESULTS: Significant correlation (intraclass correlation = 0.80 and 0.81, P < 0.001) could be detected between working length measurements obtained using Propex Pixi and Formatron D10 and measurements obtained using conventional radiography. Higher percentage of negative behaviors (negative and definitely negative) was observed during the conventional radiographic method (68.6%) as opposed to the Propex Pixi (16.5%) and Formatron D10 (20.8%) methods. CONCLUSIONS: Both the apex locators were as accurate as conventional radiography in determining working length in primary teeth; Formatron D10 being more accurate than Propex Pixi. The EALs were far better accepted by children than conventional radiography.

Anti-Ulcerative Effect of Curcumin-Galactomannoside Complex on Acetic Acid-Induced Experimental Model by Inhibiting Inflammation and Oxidative Stress.

Ulcerative colitis (UC) is a chronic inflammatory bowel disease that affects the mucosa and submucosa of colon. The pathogenesis of ulcerative colitis (UC) is related to reduced antioxidant capacity and increased inflammatory processes. Reactive oxygen metabolites are the potent inflammatory mediators that may be involved in tissue injury in inflammatory bowel disease. Conventional drug therapies for UC come with a myriad of side effects which further raise the need for natural bioactive agents. Curcumin has proven to be beneficial in the prevention and treatment of a number of inflammatory diseases, but due its poor bioavailability, the therapeutic applications are limited. Thus, to enhance its bioavailability, a new formulation - curcumin-galactomannoside (CGM)- was made by complexing curcumin with galactomannans derived from fenugreek. The present study aims to evaluate the effects of CGM on experimental UC model. Adult male Wistar rats were divided into 5 groups: normal control rats (NC); ulcerative colitis control rats (UC); UC + sulfasalazine (SS) treated; UC + curcumin (CM) treated; and UC + CGM supplemented for 21 days. The colonic mucosal injury was assessed by macroscopic and histological examination, along with evaluation of antioxidant status, inflammatory mediators, and gene expressions. Administration of CGM significantly enhanced antioxidant activities and decreased the level of inflammatory mediators and also suppressed the expression of inflammatory markers as compared with other groups. In conclusion, findings from these results reveal that CGM exerts marked curative effects on acute experimental colitis, possibly by regulating the antioxidant status and modulating inflammatory cascade.

3D Bioprinted Nanocellulose-Based Hydrogels for Tissue Engineering Applications: A Brief Review.

Nanocellulosic materials, such as cellulose nanocrystals, cellulose nanofibers, and bacterial nanocellulose, that display high surface area, mechanical strength, biodegradability, and tunable surface chemistry have attracted great attention over the last decade for biomedical applications. Simultaneously, 3D printing is revolutionizing the field of biomedical engineering, which enables the fast and on-demand printing of customizable scaffolds, tissues, and organs. Nanocellulosic materials hold tremendous potential for 3D bioprinting due to their printability, their shear thinning behavior, their ability to live cell support and owing to their excellent biocompatibility. The amalgamation of nanocellulose-based feedstocks and 3D bioprinting is therefore of critical interest for the development of advanced functional 3D hydrogels. In this context, this review briefly discusses the most recent key developments and challenges in 3D bioprinting nanocellulose-based hydrogel constructs that have been successfully tested for mammalian cell viability and used in tissue engineering applications.

Curcumin-galactomannoside complex inhibits pathogenesis in Ox-LDL-challenged human peripheral blood mononuclear cells.

Oxidised low-density lipoprotein (ox-LDL) is a pro-atherogenic molecule, which induces inflammatory response and contributes to the pathogenesis of vascular dysfunction to atherosclerosis. The aim of the present study was to explore the anti-inflammatory effect of a novel bioavailable formulation of curcumin as 'curcumagalactomannosides' (CGM) against ox-LDL-induced inflammatory responses in human peripheral blood mononuclear cells (hPBMCs). Curcumagalactomannosides was made from natural curcumin using the soluble dietary fibre (galactomannans) derived from fenugreek seeds (Trigonella foenumgracum) and the hPBMCs were isolated from healthy human volunteers. The cells were cultured in collagen-coated plates at 37 °C and grouped as Group I (Control), Group II (ox-LDL treated) and Group III (ox-LDL + CGM treated). Further analysis of inflammatory markers, reactive oxygen species and mRNA expression levels indicated significantly increased expressions of iNOS, TNF-α, IL-6 and VCAM-1 in ox-LDL-treated group along with the nuclear translocation of NF-κB. Other inflammatory markers such as LOX, PGE2, total COX and lipid peroxidation level were also found to be significantly (p < 0.05) increased upon ox-LDL treatment. The treatment with CGM on the other hand was found to down-regulate and reverse the ox-LDL-induced alterations indicating its potential anti-inflammatory effect on hPBMCs via. NF-κB signalling pathway.

A quantitative analysis of total carbohydrate content from the salivary expectorants in young children.

BACKGROUND: In this postfluoride era, the concentration of fermentable carbohydrate in saliva after food intake is important to determine the risk of developing dental caries. AIM: The aim of this study is to estimate the total carbohydrate content of salivary expectorants following consumption of commercially processed snacks. DESIGN: Thirty children aged 6-8 years were selected for estimation of total carbohydrate content of salivary expectorants using modified calorimetric anthrone-sulfuric acid-glucose reaction. The test foods analyzed were as follows: Test food A - potato chips, Test food B - glucose biscuits, Test food C - Oreo biscuits, Test food D - cake, and Test food E - cornflakes. The data obtained were analyzed using student's t-test and ANOVA. RESULTS: The difference between the mean carbohydrate values of salivary expectorants of various processed test food groups at 0 and 10 min was statistically significant (P ≤ 0.001). After 10 min interval, cornflakes were found to have highest total carbohydrate content in salivary expectorant (5.186 mg/ml). CONCLUSION: The foods with high starch content such as cornflakes and potato chips exhibited higher total carbohydrate content, thus depicting lower salivary clearance rate.

Interrogation of an autofluorescence-based method for protein fingerprinting.

In the present study, we have designed a laser-induced fluorescence (LIF) based instrumentation and developed a sensitive methodology for the effective separation, visualization, identification and analysis of proteins on a single platform. In this method, intrinsic fluorescence spectra of proteins were detected after separation on 1 or 2 dimensional Sodium Dodecyl Sulfate-Tris(2-carboxyethyl)phosphine (SDS-TCEP) polyacrylamide gel electrophoresis (PAGE) and the data were analyzed. The MATLAB assisted software was designed for the development of PAGE fingerprint for the visualization of protein after 1- and 2-dimensional protein separation. These provided objective parameters of intrinsic fluorescence intensity, emission peak, molecular weight and isoelectric point using a single platform. Further, the current architecture could differentiate the overlapping proteins in the PAGE gels which otherwise were not identifiable by conventional staining, imaging and tagging methods. Categorization of the proteins based on the presence or absence of tyrosine or tryptophan residues and assigning the corresponding emission peaks (309-356 nm) with pseudo colors allowed the detection of proportion of proteins within the given spectrum. The present methodology doesn't use stains or tags, hence amenable to couple with mass spectroscopic measurements. This technique may have relevance in the field of proteomics that is used for innumerable applications.

Anti-inflammatory effect of a novel formulation of coconut inflorescence sap against ox-LDL induced inflammatory responses in human peripheral blood mononuclear cells by modulating TLR-NF-κB signaling pathway.

Oxidized low density lipoprotein (ox LDL) induced inflammatory response was reported to play an important role in the pathogenesis of atherosclerosis. The purpose of this study was to explore the anti-inflammatory effect of a novel formulation of coconut inflorescence sap (CSP); COCOZEN™ against ox-LDL induced inflammatory responses in human peripheral blood mononuclear cells (hPBMCs). The hPBMCs were isolated from healthy human volunteers and cultured in collagen coated plates at 37 °C. The cells were grouped as Group I (Control), Group II (ox-LDL treated) and Group III (ox-LDL + CSP treated). Further analysis of inflammatory markers, reactive oxygen species, mRNA and protein expression levels indicated increased expressions of TLR-4, TNF-α, IL-6 and VCAM-1 in ox-LDL treated group along with the nuclear translocation of NF-κB. Other inflammatory markers such as LOX, PGE2, NO, total COX and lipid peroxidation level were also found to be significantly (p < .05) increased upon Ox-LDL treatment. The treatment with CSP on the other hand was found to down regulate and reverse the ox-LDL-induced alterations indicating its potential anti-inflammatory effect on hPBMCs via TLR-NF-κB signaling pathway.

Anti-inflammatory and antioxidant effect of Kerabala: a value-added ayurvedic formulation from virgin coconut oil inhibits pathogenesis in adjuvant-induced arthritis.

Kerabala (CB) is a novel ayurvedic formulation used for treating various inflammatory diseases. This formulation was made from virgin coconut oil and it comprises extracts of Sida cordifolia, coconut milk and sesame oil. The current study was performed to evaluate the anti-inflammatory action of CB on carrageenan-induced acute and adjuvant-induced chronic experimental models. 5 mg/kg bwt was found to be potent dose from carrageenan model and evaluated its effect in adjuvant-induced chronic arthritic model. The antioxidant assays like SOD, catalase, glutathione peroxidase, lipid peroxidation product, nitrate level and GSH were measured in paw tissue. Hematological parameters like hemoglobin (HB) count, ESR, WBC count, plasma CRP levels were analyzed. By RT-PCR, the inflammatory markers like cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) expressions were evaluated. The extracellular matrix proteins like MMP-2 and MMP-9 were determined by zymography and its expression by western blotting. Histopathology and cytology of paw tissue and synovium were analyzed. The result indicated that there was a significant increment in the levels of antioxidant enzymes on CB administration. The hematological markers such as ESR, WBC and plasma CRP levels were reduced by CB treatment and it also increases the HB level. The upregulated gene level expressions of inflammatory markers like COX-2, iNOS, TNF-α and IL-6 were down regulated by administration of CB. MMP-2 and MMP-9 expression significantly reduced by CB administration. Massive influx of inflammatory cell infiltration, proliferative collagen in histological analysis of paw tissue of arthritic rat was decreased by CB administration. Synovial cytology of CB administrated group shows reduced number of reactive mesothelial cells and synovial inflammatory cells. This current study shows that ayurvedic drug CB has an antioxidant, anti-inflammatory and anti-arthritic activity in experimental arthritic model. CB as an anti-arthritic drug has beneficial effect for treating inflammation, tissue damage and pain associated with arthritis.

DNA probe based colorimetric method for detection of rifampicin resistance of Mycobacterium tuberculosis.

Rifampicin resistance of Mycobacterium tuberculosis is due to the occurrence of point mutations of the rpoB gene and the site of mutations vary geographically. Commercialized molecular based methods are not able to comprehensively detect rifampicin resistance as they target a limited number of gene mutations which are thought to be common. The aim of the study was to establish a low cost DNA probe based colorimetric method that can be customized for detection of rifampicin resistance of M. tuberculosis. Thus, enzyme-linked oligosorbent assay (ELOSA) was developed for the detection of polymerase chain reaction (PCR) amplified fragments of rpoB gene of M. tuberculosis DNA on microtiter plates. Forty two M. tuberculosis isolates (rifampicin resistant and susceptible isolates identified by agar proportion method) were used for developing and validating the assay. The point mutations of resistant isolates had been previously determined by DNA sequencing. Two fragments of rpoB gene were labeled with digoxigenin by PCR. The amplified products were hybridized with selected allele specific probes for three mutations and its wild types (six probes) which were captured onto streptavidin coated microtiter plates and detected by color development. Both sensitivity and specificity of all probes were ≥96% and there was excellent discrimination (area under the curve (AUC)>0.9) between rifampicin susceptible cases and resistant cases. The probe-based colorimetric assay (PCR-ELOSA) developed in this study showed good agreement with reference mutations that were confirmed by DNA sequencing. In conclusion, PCR-ELOSA is a reliable and economical assay that can be customized for detection of rifampicin resistance.

The manual mycobacteria growth indicator tube and the nitrate reductase assay for the rapid detection of rifampicin resistance of M. Tuberculosis in low resource settings.

BACKGROUND: Tuberculosis (TB) is a disease of poverty that contributes significantly to ill-health in developing countries. Drug resistant TB is a major challenge to disease control. Early diagnosis and rapid determination of drug sensitivity is of paramount importance in eradication of TB. Although automated liquid culture based methods are available for rapid detection of drug resistance, the high cost of these tests prevent them from being used routinely in low resource settings. This study compares two phenotypic methods, the manual Mycobacteria Growth Indicator Tube (MGIT) and the Nitrate Reductase Assay (NRA) in liquid medium, with the agar proportion method (APM), the gold standard for susceptibility testing of Mycobacterium tuberculosis. METHODOLOGY: Fourteen day old M. tuberculosis strains (n=373) grown on solid media were used for drug susceptibility testing by APM, NRA and the manual MGIT method. Rifampicin free and rifampicin incorporated (final concentration, 1 µg/ml) media were inoculated with the recommended concentrations of mycobacterial suspensions and incubated at 37°C in 5% CO2. In the APM, the proportion of colonies in the drug containing medium was determined. In the NRA, the colour change in the medium was compared with a standard colour series after day 6 and day 12 of incubation. Growth in the MGIT was detected using the manual MGIT reader from day 2 onwards. The 2 methods were compared with the gold standard, APM to determine sensitivity and specificity and agreement between the methods was calculated using kappa statistics. RESULTS: Thirty one (31) rifampicin resistant isolates were identified. When compared with the APM, the sensitivity of detection of rifampicin resistance was 85% for the NRA and 93% for the manual MGIT and the specificity was 99% and 100% respectively. Both assays, NRA (κ=0.86) and manual MGIT method (κ= 0.94) were in excellent agreement with the APM. The mean turnaround time for manual MGIT method and NRA were 08 days and 10 days respectively. CONCLUSION: The NRA in liquid medium and manual MGIT are useful alternatives to APM for drug susceptibility testing of M. tuberculosis in low resource settings.

Multiple matrix metalloproteinases in type II collagen induced arthritis.

To study the relative contribution of various matrix degrading enzymes in the pathogenesis of arthritis, changes in the levels of various matrix metalloprtoteinases (MMPs) during the progression of collagen induced arthritis was studied in experimental animals. Arthritis was induced in male wistar rats by injecting an emulsion containing collagen type II and Freund's complete adjuvant. The duration of the experiment was 35 days. Synovial effusate was collected at regular intervals after induction. At the end of the experiment serum and cartilage were collected and analysed. Synovial fluid of osteoarthritic patients was also analyzed. Levels of MMP-2, MMP-3, MMP-9 and MT1-MMP were found to be high in synovial effusate and cartilage of experimental animals. In synovial effusate of arthritic animals the expression of MMP-3 was found to be high during the early stages while increase in MMP-2 and MMP-9 occurred at later stages. Synovial fluid of osteoarthritic patients also showed elevated levels of MMP-2, MMP-3 and MMP-9. Our results indicated that sequential action of MMPs such as MMP-3, MMP-2 and MMP-9 can cause degradation of articular cartilage extracellular matrix.

Effect of glycosaminoglycans on matrix metalloproteinases in type II collagen-induced experimental arthritis.

Matrix metalloproteinases (MMPs) are a family of neutral proteinases that are involved in tissue remodeling by mediating degradation of extracellular matrix components in both physiology and pathology. As MMPs appear to play a key role in the degradation of cartilage matrix in the progression of arthritic disease, MMPs are considered as potential therapeutic targets. The effect of chondroitin sulfate A (CSA) on MMPs in type II collagen-induced experimental arthritis was studied. The anti-arthritic effect of CSA was evidenced by a decrease in marker activities like lysosomal beta-hexosaminidase and beta-glucuronidase. Arthritic animals showed significantly higher activity of MMP2 and MMP9 and increased levels of other MMPs, including MMP3 and MT-1 MMP in cartilage and serum. Treatment with CSA significantly decreased the activity of MMPs, particularly MMP9 in serum and synovial effusate and cartilage. The effect of CSA was further studied by fragmenting CSA into low-molecular-weight oligosaccharides. The oligosaccharide-treated animals showed considerably lower MMP activity (particularly MMP9) compared with arthritic controls. The CSA (and the oligosaccharides derived from it) not only reduced the activity of MMPs but also decreased the protein level expression of MMPs, indicating that the production of MMPs is affected. These results indicate that the antiarthritic effect of CSA involves down-regulation of MMPs, which are critically involved in the progression of arthritic disease.

Parallel changes in fibronectin and alpha5beta1 integrin in articular cartilage in type II collagen-induced arthritis.

Interactions of cells with extracellular matrix (ECM) are mediated through specific cell surface receptors, belonging to the integrin family of transmembrane proteins. Integrins have been shown to be involved in chondrocyte-matrix interactions in the cartilage. In this study, the status of a matrix glycoprotein fibronectin (FN) and its receptor alpha5beta1 integrin in the articular cartilage in collagen type II-induced experimental arthritis in rats, as well as in synovial fluid from osteoarthritic patients was investigated. Experimental arthritis was induced by intradermal injection of type-II collagen (300 microg/100 g body wt) and Freund's complete adjuvant. Saline-treated animals served as control. Clinical severity was indicated by increase in paw volume. Significant increase in the activities of lysosomal enzymes beta-glucuronidase and beta-hexosaminidase was observed in synovial effusate, serum and cartilage of arthritic animals, when compared to untreated control, indicating dysfunction of cartilage. Changes in FN and alpha5beta1 integrin were studied by ELISA. A progressive increase was observed in the FN level in synovial effusate and cartilage of arthritic animals, when compared to untreated controls. FN levels were also significantly high in synovial fluid of osteoarthritic patients. A significant increase in the levels of alpha5beta1 integrin was found in cartilage of arthritic rats. Parallel changes in FN and alpha5beta1 integrin indicated that alterations in FN and alpha5beta1 integrin in chondrocytes constituted one of the molecular mechanisms during progression of arthritis.