Sex-specific effects of injury and beta-adrenergic activation on metabolic and inflammatory mediators in a murine model of post-traumatic osteoarthritis.

OBJECTIVE: Metabolic processes are intricately linked to the resolution of innate inflammation and tissue repair, two critical steps for treating post-traumatic osteoarthritis (PTOA). Based on lipolytic and immunoregulatory actions of norepinephrine, we hypothesized that intra-articular ß-adrenergic receptor (ßAR) stimulation would suppress PTOA-associated inflammation in the infrapatellar fat pad (IFP) and synovium. DESIGN: We used the ßAR agonist isoproterenol to perturb intra-articular metabolism 3.5 weeks after applying a non-invasive single-load compression injury to knees of 12-week-old male and female mice. We examined the acute effects of intra-articular isoproterenol treatment relative to saline on IFP histology, multiplex gene expression of synovium-IFP tissue, synovial fluid metabolomics, and mechanical allodynia. RESULTS: Injured knees developed PTOA pathology characterized by heterotopic ossification, articular cartilage loss, and IFP atrophy and fibrosis. Isoproterenol suppressed the upregulation of pro-fibrotic genes and downregulated the expression of adipose genes and pro-inflammatory genes (Adam17, Cd14, Icam1, Csf1r, and Casp1) in injured joints of female (but not male) mice. Analysis of published single-cell RNA-seq data identified elevated catecholamine-associated gene expression in resident-like synovial-IFP macrophages after injury. Injury substantially altered synovial fluid metabolites by increasing amino acids, peptides, sphingolipids, phospholipids, bile acids, and dicarboxylic acids, but these changes were not appreciably altered by isoproterenol. Intra-articular injection of either isoproterenol or saline increased mechanical allodynia in female mice, whereas neither substance affected male mice. CONCLUSIONS: Acute ßAR activation altered synovial-IFP transcription in a sex and injury-dependent manner, suggesting that women with PTOA may be more sensitive than men to treatments targeting sympathetic neural signaling pathways.

Study of chemical effects on L X-rays spectra of 59Pr compounds using high resolution WDXRF and EDXRF measurements.

Chemical effects on L X-rays spectra of PrF3, PrCl3, PrBr3, Pr2O3, Pr6O11 and Pr2(SO4)3 compounds have been investigated from measured ILi/ILα (i = l and ß6), ILj/ILß1 (j = Î·, γ5 and γ1), ILk/ILγ1 (k = Î³5 and η) intensity ratios using high resolution poly-chromatic WDXRF, and monochromatic excitation by 6.49 keV in vacuum (10-2 Torr) and Ag Kαß (22.581 keV) X-rays photons in EDXRF spectrometers. The experimental results clearly exhibit significant variation in measured intensity ratios of L X-ray components of investigated compounds from pure elemental form and theoretically predicted values evaluated using different atomic parameters. Furthermore, the change in inner shell/subshells binding energy resulting from the transitions of outer shell/subshell electrons is also inferred from the shifts of order ∼0.1-0.70 eV in Ll, Lß2 and Lγ1 components of PrF3, PrCl3, PrBr3, Pr2O3, Pr6O11 and Pr2(SO4)3 WDXRF spectra relative to pure 59Pr. The variation in intensity ratios and shift in Ll, Lß2 and Lγ1 X-ray components of 59Pr compounds are attributed to crystal defects, structural effects and exchange interactions between core and valence electrons of different ligands attached to central 59Pr atom. The reliable experimental data would be helpful in the theoretical interpretation of standard reference data for inner-shell vacancy decay parameters used in X-ray fluorescence analysis of compound samples.

Imaging mass cytometry reveals tissue-specific cellular immune phenotypes in the mouse knee following ACL injury.

Objective: To develop an imaging mass cytometry method for identifying complex cell phenotypes, inter-cellular interactions, and population changes in the synovium and infrapatellar fat pad (IFP) of the mouse knee following a non-invasive compression injury. Design: Fifteen male C57BL/6 mice were fed a high-fat diet for 8 weeks prior to random assignment to sham, 0.88 â€‹mm, or 1.7 â€‹mm knee compression displacement at 24 weeks of age. 2-weeks after loading, limbs were prepared for histologic and imaging mass cytometry analysis, focusing on myeloid immune cell populations in the synovium and IFP. Results: 1.7 â€‹mm compression caused anterior cruciate ligament (ACL) rupture, development of post-traumatic osteoarthritis, and a 2- to 3-fold increase in cellularity of synovium and IFP tissues compared to sham or 0.88 â€‹mm compression. Imaging mass cytometry identified 11 myeloid cell subpopulations in synovium and 7 in IFP, of which approximately half were elevated 2 weeks after ACL injury in association with the vasculature. Notably, two monocyte/macrophage subpopulations and an MHC IIhi population were elevated 2-weeks post-injury in the synovium but not IFP. Vascular and immune cell interactions were particularly diverse in the synovium, incorporating 8 unique combinations of 5 myeloid cell populations, including a monocyte/macrophage population, an MHC IIhi population, and 3 different undefined F4/80+ myeloid populations. Conclusions: Developing an imaging mass cytometry method for the mouse enabled us to identify a diverse array of synovial and IFP vascular-associated myeloid cell subpopulations. These subpopulations were differentially elevated in synovial and IFP tissues 2-weeks post injury, providing new details on tissue-specific immune regulation.

Fabrication of thin Molybdenum backed target using rolling method.

A successful attempt was made to fabricate a thin foil of natural Mo target on a thick Au backing with Indium in between to improve adhesion between the foils. Rolling at elevated temperature was considered to fabricate Mo foil while gold foil was fabricated employing conventional rolling technique. The heating of Mo foil under natural environment lead to the oxidation or carbonization on foil surface which was confirmed through Energy Dispersive X-ray Spectroscopy (EDS) measurements. Indium of thickness ∼86µg/cm2 was evaporated on Mo foil to improve adhesion between Mo and Au foils. The characterization of fabricated thin Mo foil was done using the Energy Dispersive X-ray Spectroscopy (EDS) and the Scanning Electron microscope (SEM) techniques. Thickness measurement of the target (Mo-Au) was done using Energy Dispersive X-ray Fluorescence (EDXRF) technique, in the measurements the thickness of the Mo foil and of gold backing are found out to be 1.3 mg/cm2 and 9 mg/cm2 respectively.

Is Old (Fludrabine/Busulfan/Cyclophosphamide/rAntiThymocyteGlobulin) Conditioning Still Gold for Allogeneic Transplants in Transfusion Dependent Beta-Thalassemia of All Risk Categories in 21st Century?

Allogeneic stem cell transplant (allo-SCT) is the only curative option for transfusion dependent thalassemia (TDT) until the gene therapy could bring paradigm shift. We analysed TDT allo-SCTs performed with Flu/Bu/Cy/rATG conditioning between October 2018 and April 2022 at our center. A retrospective analysis of 55 consecutive HLA matched alloSCT for TDT and was approved by hospital's Institutional Review Board. Median age was 7(2-13) years. On presentation, number of patients with Class I, II, III were 18 (32.7%), 14(25.4%) and 23(41.8%) respectively {ClassIIIA = 14(25.4%),ClassIIIB = 9(16.3%)}. After downstaging, Class I, II, III were 22(40%), 15(27.2%) and 18(32.7%) patients respectively {ClassIIIA = 15(27.2%),ClassIIIB = 3(5.4%)}. Graft was bone marrow in 53(96.4%) and peripheral blood stem cell in 2(3.6%) patients. Mean CD34 stem cell dose was 3.28(1.2-6.5) × 106/kg. Neutrophils and platelets engrafted at a median of 16(12-32) and 17(12-48) days. Median duration of follow-up was 20.7(1.8-43.9) months. There was no primary rejection. Although, mixed chimerism was common {17(30.9%)}, there was only one secondary rejection (1.8%). Venoocclusive disease was seen 12(21.8%) patients {mild = 9(75%), moderate = 2(16.6%) and severe = 1(8.3%)}. Acute and Chronic graft versus host disease was observed in 4(7.2%) and 4(7.2%) patients respectively. There was no treatment related mortality. Overall survival and Thalassemia Free Survival were 100% ± 0% and 98% ± 2% respectively. Flu/Bu/Cy/rATG conditioning with BM graft is a safe and effective regimen even in higher risk. It also highlights the importance of pretransplant downstaging of risk class in improving the outcomes.

Venoms classification and therapeutic uses: a narrative review.

The mere glimpse of venomous animals has always terrified humans because of the devastating effects of their venoms. However, researchers across the globe have isolated therapeutically active ingredients from these venoms and continue to explore them for drug leads. These efforts lead to the discovery of therapeutic molecules that the US-FDA has approved to treat different diseases, such as hypertension (Captopril), chronic pain (Ziconotide), and diabetes (Exenatide). The main active constituents of most venoms are proteins and peptides, which gained more attention because of advancements in biotechnology and drug delivery. The utilization of newer screening approaches improved our understanding of the pharmacological complexity of venom constituents and facilitated the development of novel therapeutics. Currently, with many venom-derived peptides undergoing different phases of clinical trials, more are in pre-clinical drug development phases. This review highlights the various sources of venoms, their pharmacological actions, and the current developments in venom-based therapeutics.

Elemental and Chemical Phase Analyses of Ras-Family Ayurvedic Medicinal Products.

Fifteen Ayurvedic medicines of Ras-family (herbo-mineral-metallic preparations) from three reputed manufactures were analysed for elemental quantification and their chemical phase identification using the energy-dispersive (ED) and wavelength-dispersive (WD) X-ray fluorescence (XRF) techniques, and powder X-ray diffraction (XRD) technique, respectively. The low-Z elements C, H, N, S and O constituting a major portion of these medicines were also determined by CHNSO analyser and further used as input for XRF analyses. The elements of concern, Hg, Pb and As, are identified in different medicine products with disquiet concentration values (maximum concentration values range ~ 4-10%) and that too with substantial variations in the products from different manufacturers. These elements are identified mainly in the cinnabar (α-HgS)/metacinnabar (ß-HgS), litharge (PbO) and alacranite (As4S4) phases in different medicines. Keeping in view the high concentration of chemicals of the Hg, Pb and As elements in the Ras-family medicines, it is vitally required to investigate their bioaccessibility and surmise the associated toxicological aspects. It is suggested that the formation of the bioaccessible toxic chemical forms of the Hg, Pb and As elements be avoided during preparation of the mineral ingredients or these soluble chemical forms be removed at suitable stage of the preparation. In view of large variations observed for the Hg, Pb and As based ingredients in the Ras family Ayurvedic medicine products from different manufacturers, adequate quality control mechanisms and production regulations are recommended.

Sirt3 Promotes Chondrogenesis, Chondrocyte Mitochondrial Respiration and the Development of High-Fat Diet-Induced Osteoarthritis in Mice.

Understanding how obesity-induced metabolic stress contributes to synovial joint tissue damage is difficult because of the complex role of metabolism in joint development, maintenance, and repair. Chondrocyte mitochondrial dysfunction is implicated in osteoarthritis (OA) pathology, which motivated us to study the mitochondrial deacetylase enzyme sirtuin 3 (Sirt3). We hypothesized that combining high-fat-diet (HFD)-induced obesity and cartilage Sirt3 loss at a young age would impair chondrocyte mitochondrial function, leading to cellular stress and accelerated OA. Instead, we unexpectedly found that depleting cartilage Sirt3 at 5 weeks of age using Sirt3-flox and Acan-CreERT2 mice protected against the development of cartilage degeneration and synovial hyperplasia following 20 weeks of HFD. This protection was associated with increased cartilage glycolysis proteins and reduced mitochondrial fatty acid metabolism proteins. Seahorse-based assays supported a mitochondrial-to-glycolytic shift in chondrocyte metabolism with Sirt3 deletion. Additional studies with primary murine juvenile chondrocytes under hypoxic and inflammatory conditions showed an increased expression of hypoxia-inducible factor (HIF-1) target genes with Sirt3 deletion. However, Sirt3 deletion impaired chondrogenesis using a murine bone marrow stem/stromal cell pellet model, suggesting a context-dependent role of Sirt3 in cartilage homeostasis. Overall, our data indicate that Sirt3 coordinates HFD-induced changes in mature chondrocyte metabolism that promote OA. © 2022 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

Genome-scale analysis of Arabidopsis splicing-related protein kinase families reveals roles in abiotic stress adaptation.

Nearly 60 - 80 % of intron-containing plant genes undergo alternative splicing in response to either stress or plant developmental cues. RNA splicing is performed by a large ribonucleoprotein complex called the spliceosome in conjunction with associated subunits such as serine arginine (SR) proteins, all of which undergo extensive phosphorylation. In plants, there are three main protein kinase families suggested to phosphorylate core spliceosome subunits and related splicing factors based on orthology to human splicing-related kinases: the SERINE/ARGININE PROTEIN KINASES (SRPK), ARABIDOPSIS FUS3 COMPLEMENT (AFC), and Pre-mRNA PROCESSING FACTOR 4 (PRP4K) protein kinases. To better define the conservation and role(s) of these kinases in plants, we performed a genome-scale analysis of the three families across photosynthetic eukaryotes, followed by extensive transcriptomic and bioinformatic analysis of all Arabidopsis thaliana SRPK, AFC, and PRP4K protein kinases to elucidate their biological functions. Unexpectedly, this revealed the existence of SRPK and AFC phylogenetic groups with distinct promoter elements and patterns of transcriptional response to abiotic stress, while PRP4Ks possess no phylogenetic sub-divisions, suggestive of functional redundancy. We also reveal splicing-related kinase families are both diel and photoperiod regulated, implicating different orthologs as discrete time-of-day RNA splicing regulators. This foundational work establishes a number of new hypotheses regarding how reversible spliceosome phosphorylation contributes to both diel plant cell regulation and abiotic stress adaptation in plants.

Comparative Appraisal of Leaf Proteomic and Mass Spectrometry Analyses During Fusarium Wilt Infection in Resistance and Susceptible Genotypes of Castor (Ricinus communis L.).

The resistant and susceptible genotypes of castor were utilized for leaf proteomic study during Fusarium wilt infection. The histopathological study was observed under SEM and it confirmed that the infection of Fusarium oxysporum f. sp. ricini was higher in the root of susceptible JI-35, while incompatible interaction is observed in resistant SKI-215 genotype. The acidic and neutral proteins were maximally up-expressed with 2 to 171 kDa in treated resistant and 2 to 150 kDa in treated susceptible interactions. In resistant genotype, the leaf proteins were recognized with 3.0- and 5.8-fold higher at infection stage and post-infection stage, respectively, as compared to susceptible genotype. The highly up expressions of leaf acidic (4.76 pI) and basic (8.77 pI) proteins were found with 224.94- and 61.68-fold change, respectively during the post-infection stage in treated resistance compared to its control. The protein spots at 4.76 pI and 8.77 pI were characterized with nanoLC-MS Triple TOF and were recognized as signalling molecules small GTP binding protein (23 kDa) and actin (8 kDa), respectively, on the basis of mass spectrometry and peptide sequences. However, basic and neutral proteins were up regulated as 30.11- and 20.30-fold changes in treated susceptible compared to its control. These proteins were identified as HSP90 (10 kDa) and LEA (27 kDa) proteins. The 148 kDa protein is recognized as histidine kinase in incompatible resistant interaction compared to compatible susceptible (serine threonine protein kinase, 65 kDa) as common acidic protein at 3.80 pI during infection stage. Some acidic proteins were maximally up-regulated in the leaf of resistant castor genotype and played a significant role in defense response.

Depigmenting effect of banana stem and flower on melanocytes.

Gingival repigmentation is an inevitable hindrance among different procedures accepted for gingival depigmentation. To overcome this, there is a need for the procedure that can delay the duration of reappearance of pigmentation. A number of studies using herbal extracts with antioxidant property shown to have anti melanogenic effect. In the present in - vitro study, we investigated the effect of banana stem and flower extracts on melanocytes, as Banana stem and flower are rich in polyphenols and flavonoids, which are potent antioxidants. The melanocytes were exposed to ethanolic extract of banana stem and flower at 2 concentrations (100 µgm, 150 µgm) for 72 h. The cellular melanin contents were measured using Bradford assay which depicted the reduction in the melanin content and Resazurin assay was used for assessment of cell viability showed no significant cytotoxic effect of banana stem and flower on the cells. The cells exposed to higher concentration (150 µgm) of banana stem and flower showed significant reduction in melanin content. Flower extract showed better reduction in the melanin content. Based on these results both banana flower and stem can be tried as potent gingival depigmenting agent.

Efficacy and safety of hydroxychloroquine for managing glycemia in type-2 diabetes: A systematic review and meta-analysis.

Aims: No Cochrane meta-analysis with grading of evidence is available on use of hydroxychloroquine (HCQ) in type-2 diabetes (T2DM). This meta-analysis evaluated the efficacy and safety of HCQ in T2DM. Methods: Electronic databases were searched using a Boolean search strategy: ((hydroxychloroquine) OR (chloroquine*)) AND ((diabetes) OR ("diabetes mellitus") OR (glycemia) OR (glucose) OR (insulin)) for studies evaluating hydroxychloroquine for glycemic control in T2DM. The primary outcome was a change in glycated haemoglobin (HbA1c). The secondary outcomes were changes in other glycemic/lipid parameters and adverse effects. Results: Data from 11 randomized controlled trials (RCTs) (3 having placebo as controls [passive controls] and 8 having anti-diabetes medications as controls [active controls]) involving 2,723 patients having a median follow-up of 24 weeks were analyzed. About 54.54% of the RCTs were of poor quality as evaluated by the Jadad scale. The performance bias and detection bias were at high risk in 63.64% of the RCTs. The HbA1c reduction with HCQ was marginally better compared to the active (mean differences [MD]-0.17% [95%, CI:-0.30--0.04;P=0.009;I2=89%; very low certainty of evidence, VLCE]), and passive (MD-1.35% [95%CI:-2.10--0.59;P=0.005;I2=74%]) controls. A reduction in fasting glucose (MD-16.63mg/dL[95%, CI: -25.99 - -7.28mg/dL;P<0.001;I2=97%;VLCE]) and post-prandial glucose [MD -8.41mg/dL (95%CI: -14.71 - -2.12mg/dL;P=0.009;I2=87%;VLCE]), appeared better with HCQ compared to active controls. The total adverse events (risk ratio [RR]0.93 [95% CI:0.68-1.28]; P=0.65;I2=66%) were not different with HCQ compared to the controls. Conclusion: The routine use of HCQ in T2DM cannot be recommended based on the current evidence.

Genetic overlap analysis of endometriosis and asthma identifies shared loci implicating sex hormones and thyroid signalling pathways.

STUDY QUESTION: Is there a shared genetic or causal association of endometriosis with asthma or what biological mechanisms may underlie their potential relationships? SUMMARY ANSWER: Our results confirm a significant but non-causal association of endometriosis with asthma implicating shared genetic susceptibility and biological pathways in the mechanisms of the disorders, and potentially, their co-occurrence. WHAT IS KNOWN ALREADY: Some observational studies have reported a pattern of co-occurring relationship between endometriosis and asthma; however, there is conflicting evidence and the aetiology, as well as the underlying mechanisms of the relationship, remain unclear. STUDY DESIGN, SIZE, DURATION: We applied multiple statistical genetic approaches in the analysis of well-powered, genome-wide association study (GWAS) summary data to comprehensively assess the relationship of endometriosis with asthma. Endometriosis GWAS from the International Endogene Consortium (IEC, 17 054 cases and 191 858 controls) and asthma GWAS from the United Kingdom Biobank (UKB, 26 332 cases and 375 505 controls) were analysed. Additional asthma data from the Trans-National Asthma Genetic Consortium (TAGC, 19 954 cases and 107 715 controls) were utilized for replication testing. PARTICIPANTS/MATERIALS, SETTING, METHODS: We assessed single-nucleotide polymorphism (SNP)-level genetic overlap and correlation between endometriosis and asthma using SNP effect concordance analysis (SECA) and linkage disequilibrium score regression analysis (LDSC) methods, respectively. GWAS meta-analysis, colocalization (GWAS-PW), gene-based and pathway-based functional enrichment analysis methods were applied, respectively, to identify SNP loci, genomic regions, genes and biological pathways shared by endometriosis and asthma. Potential causal associations between endometriosis and asthma were assessed using Mendelian randomization (MR) methods. MAIN RESULTS AND THE ROLE OF CHANCE: SECA revealed significant concordance of SNP risk effects across the IEC endometriosis and the UKB asthma GWAS. Also, LDSC analysis found a positive and significant genetic correlation (rG = 0.16, P = 2.01 × 10-6) between the two traits. GWAS meta-analysis of the IEC endometriosis and UKB asthma GWAS identified 14 genome-wide significant (Pmeta-analysis < 5.0 × 10-8) independent loci, five of which are putatively novel. Three of these loci were consistently replicated using TAGC asthma GWAS and reinforced in colocalization and gene-based analyses. Additional shared genomic regions were identified in the colocalization analysis. MR found no evidence of a significant causal association between endometriosis and asthma. However, combining gene-based association results across the GWAS for endometriosis and asthma, we identified 17 shared genes with a genome-wide significant Fisher's combined P-value (FCPgene) <2.73 × 10-6. Additional analyses (independent gene-based analysis) replicated evidence of gene-level genetic overlap between endometriosis and asthma. Biological mechanisms including 'thyroid hormone signalling', 'abnormality of immune system physiology', 'androgen biosynthetic process' and 'brain-derived neurotrophic factor signalling pathway', among others, were significantly enriched for endometriosis and asthma in a pathway-based analysis. LARGE SCALE DATA: The GWAS for endometriosis data were sourced from the International Endogen Consortium (IEC) and can be accessed by contacting the consortium. The GWAS data for asthma are freely available online at Lee Lab (https://www.leelabsg.org/resources) and from the Trans-National Asthma Genetic Consortium (TAGC). LIMITATIONS, REASONS FOR CAUTION: Given we analysed GWAS datasets from mainly European populations, our results may not be generalizable to other ancestries. WIDER IMPLICATIONS OF THE FINDINGS: This study provides novel insights into mechanisms underpinning endometriosis and asthma, and potentially their observed relationship. Findings support a co-occurring relationship of endometriosis with asthma largely due to shared genetic components. Agents targeting 'selective androgen receptor modulators' may be therapeutically relevant in both disorders. Moreover, SNPs, loci, genes and biological pathways identified in our study provide potential targets for further investigation in endometriosis and asthma. STUDY FUNDING/COMPETING INTEREST(S): National Health and Medical Research Council (NHMRC) of Australia (241,944, 339,462, 389,927, 389,875, 389,891, 389,892, 389,938, 443,036, 442,915, 442,981, 496,610, 496,739, 552,485, 552,498, 1,026,033 and 1,050,208), Wellcome Trust (awards 076113 and 085475) and the Lundbeck Foundation (R102-A9118 and R155-2014-1724). All researchers had full independence from the funders. Authors do not have any conflict of interest.

Evaluation of Galectin Binding by Surface Plasmon Resonance.

Surface plasmon resonance (SPR) instruments, like the BIAcore 3000, are useful for studying the binding between macromolecules in real time. The high sensitivity and low sample consumption in the Biacore enables the measurement of rapid kinetics and low affinities characteristics of many biological interactions. This chapter describes the affinity measurement of Galectins-1, -2 and -3 and their glycoside ligands using this approach.

Fundamentals of OA. An initiative of Osteoarthritis and Cartilage. Obesity and metabolic factors in OA.

OBJECTIVE: Obesity was once considered a risk factor for knee osteoarthritis (OA) primarily for biomechanical reasons. Here we provide an additional perspective by discussing how obesity also increases OA risk by altering metabolism and inflammation. DESIGN: This narrative review is presented in four sections: 1) metabolic syndrome and OA, 2) metabolic biomarkers of OA, 3) evidence for dysregulated chondrocyte metabolism in OA, and 4) metabolic inflammation: joint tissue mediators and mechanisms. RESULTS: Metabolic syndrome and its components are strongly associated with OA. However, evidence for a causal relationship is context dependent, varying by joint, gender, diagnostic criteria, and demographics, with additional environmental and genetic interactions yet to be fully defined. Importantly, some aspects of the etiology of obesity-induced OA appear to be distinct between men and women, especially regarding the role of adipose tissue. Metabolomic analyses of serum and synovial fluid have identified potential diagnostic biomarkers of knee OA and prognostic biomarkers of disease progression. Connecting these biomarkers to cellular pathophysiology will require future in vivo studies of joint tissue metabolism. Such studies will help reveal when a metabolic process or a metabolite itself is a causal factor in disease progression. Current evidence points towards impaired chondrocyte metabolic homeostasis and metabolic-immune dysregulation as likely factors connecting obesity to the increased risk of OA. CONCLUSIONS: A deeper understanding of how obesity alters metabolic and inflammatory pathways in synovial joint tissues is expected to provide new therapeutic targets and an improved definition of "metabolic" and "obesity" OA phenotypes.

Coronally Advanced Flap Combined With Sticky Bone and i-PRF-Coated Collagen Membrane to Treat Single Maxillary Gingival Recessions: Case Series.

INTRODUCTION: Numerous techniques have been reported in the literature for the reconstruction of gingival recession defects. The purpose of this case series was to evaluate clinically and radiographically the efficacy of sticky bone with i-PRF-coated collagen membrane in the treatment of gingival recession. CASE SERIES: Sixteen patients exhibiting isolated Miller's Class I or II recession in the maxillary esthetic zone were treated using sticky bone (i-PRF + freeze-dried bone allograft) with i-PRF-coated collagen membrane using the coronally advanced flap. Clinical parameters including probing depth (PD), width of keratinized gingiva (WKG), gingival thickness (GT), and recession depth (RD) were recorded at baseline and 6 months post-surgery. The radiographic (ST-CBCT) measurements computed were labial plate thickness (OT1, OT3, and OT5) and GT (GT1, GT3, and GT5) at baseline and 6 months post-treatment. Twelve out of sixteen treated cases achieved complete root coverage. An increase in GT was observed in all the cases. CONCLUSIONS: Within the limitations of this case series, sticky bone with i-PRF-coated collagen membrane showed promising results in the treatment of isolated maxillary Miller's Class I or II gingival recession and serves as an altered approach for root coverage procedure. However, histological analysis and larger sample size are needed to establish definitive proof of soft and hard tissue regeneration.

A clinico-microbiological and biochemical study evaluating the adjunctive use of antimicrobial photodynamic therapy and local drug delivery of 1.2 % simvastatin gel compared to scaling and root planing alone.

BACKGROUND: To evaluate and compare the efficacy of antimicrobial Photodynamic therapy (aPDT), Local Drug Delivery (LDD) of 1.2 % Simvastatin gel as an adjunct to scaling and root planning (SRP) and SRP alone in the treatment of Periodontitis using clinical, microbiological and biochemical parameters. MATERIALS AND METHODS: A total of 33 test sites in 11 Grade A Stage II periodontitis patients were randomly divided into three groups: GROUP I: Treated by SRP alone (SRP group); GROUP II: Treated by SRP followed by aPDT (aPDT group); GROUP III: Treated by SRP followed by single subgingival application of 1.2 % simvastatin gel (SMV group). Clinical parameters including API, PBI, PPD and RAL were assessed. Quantification of Porphyromonas gingivalis was evaluated by RT -PCR technique and estimation of RANKL levels was checked by ELISA. All assessments were done at baseline and 3 months RESULTS: All three groups showed significant reduction in the scores of clinical parameters, P. gingivalis DNA copy numbers and GCF RANKL levels at 3 months post therapy compared to baseline (p < 0.05). On comparison between the three groups, the results were non significant for all parameters both at baseline and at 3 months post therapy (p > 0.05). However slightly greater reduction was seen in the mean scores of PPD and RAL in the SMV Group and in P. gingivalis DNA copy numbers and GCF RANKL levels in aPDT group compared to the other groups although statistically non significant. A significant positive correlation(p < 0.05) was observed between P. gingivalis DNA copy numbers and PPD scores in SMV group and a significant negative correlation(p < 0.05) was observed between P. gingivalis DNA copy numbers and API & PBI scores in SRP group at 3 months follow up. CONCLUSIONS: aPDT, 1.2 % SMV local drug delivery as adjunct to SRP and SRP alone are effective in improving clinical parameters, reducing P. gingivalis DNA copy numbers and GCF RANKL levels. The superiority of one over another modality of treatment could not be established in this short term study.

Knowledge, attitude, and implementation of nicotine replacement therapy by dental and medical interns in Davangere city: A cross-sectional survey.

BACKGROUND: One of the prime causes of illness and premature death is smoking. Almost 50% of smokers attempt to quit the habit; however, at most, 2%-3% achieve success. The rationale is that innumerable withdrawal attempts are unplanned, and the most effective cessation aids are unacquainted. Nicotine replacement therapy (NRT) is the most common cessation aid. Furthermore, motivation from dental and medical professionals can be effective for patients to quit smoking. The study aimed to assess the knowledge, attitude, and practice regarding the implementation of NRT among dental and medical interns in Davangere city. MATERIALS AND METHODS: A questionnaire-based survey was conducted, which included 442 dental and medical interns from two dental and two medical colleges in Davangere city, Karnataka. The questionnaire included multiple-choice questions regarding knowledge, attitude, and implementation of NRT. The response rate of interns was 93.67%. RESULTS: Among dental and medical interns, there was no statistically significant difference in knowledge about NRT with P = 0.976 (P > 0.05). However, a statistically significant difference existed regarding attitude and implementation in the interns about NRT among dental and medical interns with P = 0.001 (P < 0.05). Among dental and medical interns, dental interns had a positive attitude and implementation toward NRT than medical interns. CONCLUSION: The overview implicated that the dental interns had better vision than medical interns; however, both the groups' comprehension concerning NRT is scanty and advocates education about the fundamentals of NRT either via workshop or by continuing dental education programs.

Ile73Asn mutation in protein C introduces a new N-linked glycosylation site on the first EGF-domain of protein C and causes thrombosis.

Activated protein C exerts its anticoagulant activity by protein S-dependent inactivation of factors Va and VIIIa by limited proteolysis. We identified a venous thrombosis patient who has plasma protein C antigen level of 63% and activity levels of 44% and 23%, as monitored by chromogenic and clotting assays. Genetic analysis revealed the proband carries compound heterozygous mutations (c.344T>A, p.I73N and c.1181G>A, p.R352Q) in PROC We individually expressed protein C mutations and discovered that thrombin-thrombomodulin activates both variants normally and the resulting activated protein C mutants exhibit normal amidolytic and proteolytic activities. However, while protein S-dependent catalytic activity of activated protein C-R352Q toward factor Va was normal, it was significantly impaired for activated protein C-I73N. These results suggest that the Ile to Asn substitution impairs interaction of activated protein C-I73N with protein S. This conclusion was supported by a normal anticoagulant activity for activated protein C-I73N in protein S-deficient but not in normal plasma. Further analysis revealed Ile to Asn substitution introduces a new glycosylation site on first EGF-like domain of protein C, thereby adversely affecting interaction of activated protein C with protein S. Activated protein C-R352Q only exhibited reduced activity in sub-physiological concentrations of Na+ and Ca2+, suggesting that this residue contributes to metal ion-binding affinity of the protease, with no apparent adverse effect on its function in the presence of physiological levels of metal ions. These results provide insight into the mechanism by which I73N/R352Q mutations in activated protein C cause thrombosis in proband carrying this compound heterozygous mutation.