Salivary and serum estrogen level assessment in oral lichen planus patients and its correlative analysis with OLP and stress.

Background: Lichen planus is a chronic inflammatory disease of the skin and mucous membrane with higher predilection seen in the female population. Oral lichen planus (OLP) has been associated with various etiological factors, such as stress, hormonal imbalance, and immunological variation. The purpose of this study was to assess serum and salivary estrogen (E2) levels in OLP patients and correlate them with stress levels. Objectives: This study aimed to evaluate serum and salivary estrogen levels in female patients with OLP, along with the assessment of stress and its correlation with estrogen levels. Methods: A total of 78 females, 39 clinically diagnosed with OLP and 39 healthy females, were included in the study as the case and control groups, respectively. 2 ml each of salivary and serum samples was obtained from each participant to measure the estrogen levels. Stress levels in the study group patients were assessed using the Depression Anxiety Stress Scale (DASS-21) and the Perceived Stress Scale (PSS). The nonparametric Mann-Whitney test was used for intergroup comparisons. Results: Significantly higher serum estrogen levels with higher DASS-21 and PSS scores were noted in patients with OLP. Overall, significant positive correlations were observed between salivary E2 and serum E2 (r = 0.361, P = 0.001). There was a positive correlation between salivary and serum E2 and DASS score (r = 0.410, P < 0.001, and r = 0.768, P < 0.001, respectively), serum/salivary E2 and PSS score (r = 0.745, P < 0.001, and r = 0.410, P < 0.001, respectively), and DASS score and PSS score (r = 0.878, P < 0.001). Conclusion: Estrogen can be used as a useful biomarker for OLP in the future. Salivary samples can prove to be an accurate and feasible alternative to serum estrogen level determination. We also suggest that OLP patients must be given supportive psychological treatment for improved life quality and disease management.

Serum and salivary interleukin-1ß level in oral precancer: An observational study.

BACKGROUND AND AIM: Precancer biomarkers help in early detection and management of oral potentially malignant disorders (OPMDs). Interleukin-1ß (IL-1ß), a biomarker, is known to be altered in oral submucous fibrosis (OSMF) and oral leukoplakia (OL). Therefore, we evaluated and compared the serum and salivary IL-1ß levels in patients with OSMF/oral leukoplakia and in gender- and age-matched healthy individuals. MATERIALS AND METHODS: An in vivo, prospective, observational study was conducted on 40 subjects. Subjects were divided into two groups with 20 individuals in each group, that is, Group I: OSMF/oral leukoplakia and Group II: control group. Salivary and serum IL-1ß levels were quantitatively estimated using enzyme-linked immunosorbent assay (ELISA). The statistical tests used were unpaired t-test and Chi-square test. RESULTS: The serum IL-1ß levels were significantly (P 0.001) lesser in Group I in comparison to Group II. The salivary IL-1ß levels remained insignificant between both the groups. However, in both the groups, the salivary IL-1ß levels were significantly higher compared to the serum IL-1ß levels. CONCLUSION: We found that the serum IL-1ß level can be considered as a prospective biomarker for dysplasia, whereas salivary IL-1ß alone needs more elaborated studies to account for its application as a potential biomarker in OPMD.

Deciphering the Mechanisms and Biotechnological Implications of Nanoparticle Synthesis Through Microbial Consortia.

Nanomaterial synthesis is a growing study area because of its extensive range of uses. Nanoparticles' high surface-to-volume ratio and rapid interaction with various particles make them appealing for diverse applications. Traditional physical and chemical methods for creating metal nanoparticles are becoming outdated because they involve complex manufacturing processes, high energy consumption, and the formation of harmful by-products that pose major dangers to human health and the environment. Therefore, there is an increasing need to find alternative, cost-effective, dependable, biocompatible, and environmentally acceptable ways of producing nanoparticles. The process of synthesizing nanoparticles using microbes has become highly intriguing because of their ability to create nanoparticles of varying sizes, shapes, and compositions, each with unique physicochemical properties. Microbes are commonly used in nanoparticle production because they are easy to work with, can use low-cost materials, such as agricultural waste, are cheap to scale up, and can adsorb and reduce metal ions into nanoparticles through metabolic activities. Biogenic synthesis of nanoparticles provides a clean, nontoxic, ecologically friendly, and sustainable method using renewable ingredients for reducing metals and stabilizing nanoparticles. Nanomaterials produced by bacteria can serve as an effective pollution control method due to their many functional groups that can effectively target contaminants for efficient bioremediation, aiding in environmental cleanup. At the end of the paper, we will discuss the obstacles that hinder the use of biosynthesized nanoparticles and microbial-based nanoparticles. The paper aims to explore the sustainability of microorganisms in the burgeoning field of green nanotechnology.

Biofortification, metabolomic profiling and quantitative analysis of vitamin B12 enrichment in guava juice via lactic acid fermentation using Levilactobacillus brevis strain KU15152.

BACKGROUND: Chemical fortification and dose supplementation of vitamin B12 are widely implemented to combat deficiency symptoms. However, in situ, fortification of vitamin B12 in food matrixes can be a promising alternative to chemical fortification. The present study aimed to produce vitamin B12-rich, probiotic guava juice fermented with Levilactobacillus brevis strain KU15152. Pasteurized fresh guava juice was inoculated with 7.2 log CFU mL-1 L. brevis strain KU15152 and incubated for 72 h at 37 °C anaerobically. The antioxidants, total phenolic compounds, vitamin B12 production, sugars, organic acids, pH and viable count were analyzed at 24, 48 and 72 h of incubation. The fermented juice was stored at 4 °C, and the changes in its functional properties were analyzed at 7-day intervals up to 28 days of storage. RESULTS: During fermentation, the bacteria cell count was increased from 7.01 ± 0.06 to 9.76 ± 0.42 log CFU mL-1 after 72 h of fermentation and was decreased to 6.94 ± 0.34 CFU mL-1 during storage at 4 °C after 28 days. The pH, total soluble solids, crude fiber, citric acid and total sugars decreased, while titratable acidity, total protein, antioxidants, phenolic compounds and lactic acid contents increased during fermentation. The fermented guava juice exhibited higher 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS)) radical scavenging activities (85.97% and 75.97%, respectively) at 48 h of fermentation. The concentration of active vitamin B12 in the sample reached 109.5 µg L-1 at 72 h of fermentation. However, this concentration gradually decreased to 70.2 µg L-1 during the storage period. During storage for 28 days at 4 °C, both the fermented and control guava juices exhibited a decline in antioxidant and phenolic compound concentrations. Furthermore, the addition of 20% honey and guava flavor enhanced the organoleptic properties and acceptability of fermented guava juice. CONCLUSION: The value-added fermented guava juice could be a novel functional food product to combat vitamin B12 deficiency. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Autologous Concentrate Growth Factor for the Treatment of Endo-Perio Lesion - A Case Report.

INTRODUCTION: The success of a combined periodontal and endodontic lesion depends on the elimination of both these disease processes. In the case of a combined endo-perio lesion, endodontic therapy results in healing of the endodontic component of involvement, while the prognosis of teeth would finally depend on the healing of the periodontal structure. TREATMENT: This case report evaluates the efficacy of autologous fibrin glue and bone graft, that is, sticky bone in the management of bone defects associated with endo-perio lesion. The endo-perio lesion is first treated endodontically, followed by periodontal therapy. Conclusion: The patient was kept on follow-up for 9 months, and satisfactory results in terms of bone fill and reduction in pocket depth were obtained. TAKEAWAY LESSONS: The sticky bone enhances regeneration in treatment of endo-perio lesions.

Xanthan gum-capped Chromia Nanoparticles (XG-CrNPs): A promising nanoprobe for the detection of heavy metal ions.

The present study reports on the selective and sensitive detection of metals using xanthan gum-capped chromia nanoparticles (XG-CrNPs). The nanoparticles were synthesized by the chemical reduction method using sodium borohydride and xanthan gum as the reducing and capping agents, respectively. The synthesis of XG-CrNPs was confirmed by the appearance of the two absorption peaks at 272 nm and 371 nm in the UV-visible region. The nanoparticles have been extensively characterized by FTIR, TEM-EDX, XRD, and TGA analyses. The well-dispersed XG-CrNPs exhibited a quasi-spherical structure with an average particle size of 3 nm. A significantly low amount (2 µg/L) of XG-CrNPs was used for selective and sensitive detection of heavy metal ions. It showed excellent metal detecting properties by quenching its band gap signal which was extraordinarily conspicuous for Co(II), Hg(II), and Cd(II) in comparison to other metal ions like Ag(I), Ba(II), Mg(II), Mn(II), Ni(II), and Zn(II). The limit of detection of Co(II), Cd(II), and Hg(II) with this nanoprobe was found to be 2.167 µM, 1.065 µM, and 0.601 µM respectively. The nanoparticles manifested higher shelf-life and can be reused up to three consecutive cycles where most of its activity was conserved even after being used. Thus, it may find use in metal sensor devices for the detection of hazardous metals.

Remediation of Dyes Using Supramolecular Material Derived from Carbohydrate Based π-Gelator Using the Bottom-Up Assembly Approach.

As a consequence of rapid population growth, the earth has faced numerous environmental sustainability issues and crises, water pollution is one of the important points of concern because of industrialization. In particular, effluents discharged from dying industries are rated top among the various industrial effluents, especially by their volume and composition. Annually >7.5 × 105 metric tons of different dyes are produced and consumed in different industries. In order to dye 1 kg of fabric, approximately 100-150 L of water is required, and after the dying process, it is discharged as an effluent either on a landfill or in water bodies. It is our responsibility to conserve environmental sustainability. In this line, we have developed a simple protocol to generate carbohydrate-based amphiphile using D-sorbitol, and pyrene-1-carboxaldehyde in good yield. This carbohydrate-based π-gelator is prone to forming a gel in various solvents and oils by the bottom-up assembly process. Morphological analysis of the self-assembled structure was identified by using optical microscopy and SEM. The viscoelastic behavior of the gel was examined by using rheology. In this paper, we explored the dye adsorption and desorption characteristics of the gel. Further, we have developed a cartridge based on cellulose using a template-assisted assembly phenomenon and demonstrated its potential in adsorbing dyes such as methylene blue, crystal violet, rhodamine B, and Congo red.

Association of CC-chemokine ligand-2 gene polymorphisms with leprosy reactions.

BACKGROUND: C-C motif chemokine ligand 2, a gene that codes for a protein involved in inflammation. Certain SNPs in the CCL2 gene have been studied for their potential associations with susceptibility to various diseases. These SNPs may affect the production and function of the CCL2 protein, which is involved in the recruitment of immune cells to the site of inflammation. Variations in CCL2 may influence the immune response to Mycobacterium leprae infection. OBJECTIVE: To investigate the association of the C-C motif chemokine ligand-2 single nucleotide polymorphisms with leprosy. METHODS: CCL2 single nucleotide polymorphisms were analyzed in a total of 975 leprosy patients and 357 healthy controls. Of those, 577 leprosy and 288 healthy controls were analyzed by PCR-RFLP for CCL2 -2518 A>G, 535 leprosy and 290 controls for CCL2 -362 G>C, 295 leprosy and 240 controls for CCL2 -2134 T>G, 325 leprosy and 288 controls for CCL2 -1549 A>T SNPs by melting curve analysis using hybridization probe chemistry and detection by fluorescence resonance energy transfer (FRET) technique in Realtime PCR. The levels of CCL2, IL-12p70, IFN-γ, TNF-α, and TGF-ß were estimated in sera samples and correlated with CCL2 genotypes. RESULTS: The frequency of the GCT (-2518 A>G, -362 G>C, -2134 T>G) haplotype is observed to be higher in leprosy patients compared to healthy controls (P = 0.04). There was no significant difference observed in genotypic frequencies between leprosy patients and healthy controls {(-2518A>G, p = 0.53), (-362 G>C, p = 0.01), (-2134 T>G, p = 0.10)}. G allele at the -2134 site is predominant in leprosy (borderline) without any reaction (8 %) compared to borderline patients with RR reactions (2.1 %) (P = 0.03). GG genotype (p = 0.008) and G allele at -2518 (p = 0.030) of the CCL 2 gene were found to be associated with patients with ENL reaction. An elevated level of serum CCL2 was observed in leprosy patients with the -2518 AA and AG genotypes (p = 0.0001). CONCLUSIONS: G allele and GG genotype at the CCL2 -2518 site are associated with a risk of ENL reactions.

CRISPR-Cas9: A Potent Gene-editing Tool for the Treatment of Cancer.

The prokaryotic adaptive immune system has clustered regularly interspaced short palindromic repeat. CRISPR-associated protein (CRISPR-Cas) genome editing systems have been harnessed. A robust programmed technique for efficient and accurate genome editing and gene targeting has been developed. Engineered cell therapy, in vivo gene therapy, animal modeling, and cancer diagnosis and treatment are all possible applications of this ground-breaking approach. Multiple genetic and epigenetic changes in cancer cells induce malignant cell growth and provide chemoresistance. The capacity to repair or ablate such mutations has enormous potential in the fight against cancer. The CRISPR-Cas9 genome editing method has recently become popular in cancer treatment research due to its excellent efficiency and accuracy. The preceding study has shown therapeutic potential in expanding our anticancer treatments by using CRISPR-Cas9 to directly target cancer cell genomic DNA in cellular and animal cancer models. In addition, CRISPR-Cas9 can combat oncogenic infections and test anticancer medicines. It may design immune cells and oncolytic viruses for cancer immunotherapeutic applications. In this review, these preclinical CRISPRCas9- based cancer therapeutic techniques are summarised, along with the hurdles and advancements in converting therapeutic CRISPR-Cas9 into clinical use. It will increase their applicability in cancer research.

Bioremediation of hydrocarbon by co-culturing of biosurfactant-producing bacteria in microbial fuel cell with Fe2O3-modified anode.

The most common type of environmental contamination is petroleum hydrocarbons. Sustainable and environmentally friendly treatment strategies must be explored in light of the increasing challenges of toxic and critical wastewater contamination. This paper deals with the bacteria-producing biosurfactant and their employment in the bioremediation of hydrocarbon-containing waste through a microbial fuel cell (MFC) with Pseudomonas aeruginosa (exoelectrogen) as co-culture for simultaneous power generation. Staphylococcus aureus is isolated from hydrocarbon-contaminated soil and is effective in hydrocarbon degradation by utilizing hydrocarbon (engine oil) as the only carbon source. The biosurfactant was purified using silica-gel column chromatography and characterised through FTIR and GCMS, which showed its glycolipid nature. The isolated strains are later employed in the MFCs for the degradation of the hydrocarbon and power production simultaneously which has shown a power density of 6.4 W/m3 with a 93% engine oil degradation rate. A biogenic Fe2O3 nanoparticle (NP) was synthesized using Bambusa arundinacea shoot extract for anode modification. It increased the power output by 37% and gave the power density of 10.2 W/m3. Thus, simultaneous hydrocarbon bioremediation from oil-contamination and energy recovery can be achieved effectively in MFC with modified anode.

Characterization, Antioxidant, and Antimicrobial Properties of Mulberry Lattices.

In order to find the most advantageous bioactive compounds from mulberry latex for drug development in the near future, this study was conducted to characterize and evaluate antioxidant and antimicrobial properties from four different mulberry lattices (BR-2, S-1, AR-14, and S-146). The characterization of the lattices was performed by scanning electron microscopy with energy-dispersive X-ray spectroscopy, gas chromatography coupled to mass spectroscopy, and Fourier transform infrared spectroscopy. Further, screenings of the antioxidant and antimicrobial potential of selected lattices were performed in vitro using 2,2-diphenyl-1-picrylhydrazyl assay and agar well diffusion methods, respectively. Interestingly, the outcome of the current study revealed that tested mulberry lattices contain a considerable amount of bioactive phytoconstituents, particularly antimicrobial and antioxidant compounds, as revealed by chromatographic analysis. BR-2 latex was found to have significant antioxidant activity (75%) followed by S-146 (64.6%) and AR-14 (52.9%). The maximum antimicrobial activity was found in BR-2 latex compared to other tested latex varieties. The results of this investigation showed that mulberry latex from the BR-2 type may successfully control both bacterial and fungal infections, with the added benefit of having enhanced antioxidant capabilities.

Association of Early Pregnancy Values of Glycosylated Hemoglobin and the Development of Gestational Diabetes Mellitus.

Introduction There is no consensus regarding screening and diagnostic methods for gestational diabetes mellitus (GDM). The present study aimed to evaluate the association between early pregnancy values of glycosylated hemoglobin and the development of gestational diabetes mellitus among pregnant women in a tertiary care hospital in eastern India. Methods The prospective cohort study included 200 pregnant women aged between 18 and 35 years in their first trimester (gestational age eight to 13 weeks) attending the antenatal clinics of the study hospital. A glycated hemoglobin (HbA1c) test and a 75-g oral glucose tolerance test (OGTT) test were done in all study participants in their first trimester. Pregnant women with HbA1c ≥6.5% and OGTT ≥140 mg/dl were excluded from the study. In other women, the second trimester (24-28 weeks) and the third trimester OGTT (32-34 weeks) were done to detect gestational diabetes mellitus. Data collection was initiated after the approval of the Information, Education, and Communication (IEC) and relevant authorities. Receiver operating characteristic (ROC) analysis was done to identify the cut-off value of HbA1c that predicted the development of GDM. Results The incidence of GDM was 33% among our study participants. The mean HbA1c was significantly higher among women who had GDM (5.4 ± 0.4%) as compared to those who did not develop GDM (4.9 ± 0.2%) (p<0.001). On ROC analysis of HbA1c values to predict the development of GDM, a cut-off value of HbA1c ≥5.25%, irrespective of risk status, was calculated to have 84.8% sensitivity and 62.7% specificity, and among the high-risk group, HbA1c ≥5.15% had 83.3% sensitivity and 97% specificity in predicting GDM. On stratified analysis, a moderately strong positive correlation was demonstrated between HbA1c values and OGTT in the second trimester in both high-risk and low-risk cohorts (p<0.05). Conclusion Based on the findings of the present study, HbA1c can be proposed to be a suitable biomarker for GDM prediction, probably not independently but rather as a component of a multi-marker approach for high- and low-risk pregnant groups.

Use of biogenic silver nanoparticles on the cathode to improve bioelectricity production in microbial fuel cells.

To date, research on microbial fuel cells (MFCs) has. focused on the production of cost-effective, high-performance electrodes and catalysts. The present study focuses on the synthesis of silver nanoparticles (AgNPs) by Pseudomonas sp. and evaluates their role as an oxygen reduction reaction (ORR) catalyst in an MFC. Biogenic AgNPs were synthesized from Pseudomonas aeruginosa via facile hydrothermal synthesis. The physiochemical characterization of the biogenic AgNPs was conducted via scanning electron microscopy (SEM), X-ray diffraction (XRD), and UV-visible spectrum analysis. SEM micrographs showed a spherical cluster of AgNPs of 20-100 nm in size. The oxygen reduction reaction (ORR) ability of the biogenic AgNPs was studied using cyclic voltammetry (CV). The oxygen reduction peaks were observed at 0.43 V, 0.42 V, 0.410 V, and 0.39 V. Different concentrations of biogenic AgNPs (0.25-1.0 mg/cm2) were used as ORR catalysts at the cathode in the MFC. A steady increase in the power production was observed with increasing concentrations of biogenic AgNPs. Biogenic AgNPs loaded with 1.0 mg/cm2 exhibited the highest power density (PDmax) of 4.70 W/m3, which was approximately 26.30% higher than the PDmax of the sample loaded with 0.25 mg/cm2. The highest COD removal and Coulombic efficiency (CE) were also observed in biogenic AgNPs loaded with 1.0 mg/cm2 (83.8% and 11.7%, respectively). However, the opposite trend was observed in the internal resistance of the MFC. The lowest internal resistance was observed in a 1.0 mg/cm2 loading (87 Ω), which is attributed to the high oxygen reduction kinetics at the surface of the cathode by the biogenic AgNPs. The results of this study conclude that biogenic AgNPs are a cost-effective, high-performance ORR catalyst in MFCs.

Hyperglycemia is associated with duodenal dysbiosis and altered duodenal microenvironment.

The gut microbiome influences the pathogenesis and course of metabolic disorders such as diabetes. While it is likely that duodenal mucosa associated microbiota contributes to the genesis and progression of increased blood sugar, including the pre-diabetic stage, it is much less studied than stool. We investigated paired stool and duodenal microbiota in subjects with hyperglycemia (HbA1c ≥ 5.7% and fasting plasma glucose > 100 mg/dl) compared to normoglycemic. We found patients with hyperglycemia (n = 33) had higher duodenal bacterial count (p = 0.008), increased pathobionts and reduction in beneficial flora compared to normoglycemic (n = 21). The microenvironment of duodenum was assessed by measuring oxygen saturation using T-Stat, serum inflammatory markers and zonulin for gut permeability. We observed that bacterial overload was correlated with increased serum zonulin (p = 0.061) and higher TNF-α (p = 0.054). Moreover, reduced oxygen saturation (p = 0.021) and a systemic proinflammatory state [increased total leukocyte count (p = 0.031) and reduced IL-10 (p = 0.015)] characterized the duodenum of hyperglycemic. Unlike stool flora, the variability in duodenal bacterial profile was associated with glycemic status and was predicted by bioinformatic analysis to adversely affect nutrient metabolism. Our findings offer new understanding of the compositional changes in the small intestine bacteria by identifying duodenal dysbiosis and altered local metabolism as potentially early events in hyperglycemia.

A Click Chemistry-Based Artificial Metallo-Nuclease.

Artificial metallo-nucleases (AMNs) are promising DNA damaging drug candidates. Here, we demonstrate how the 1,2,3-triazole linker produced by the Cu-catalysed azide-alkyne cycloaddition (CuAAC) reaction can be directed to build Cu-binding AMN scaffolds. We selected biologically inert reaction partners tris(azidomethyl)mesitylene and ethynyl-thiophene to develop TC-Thio, a bioactive C3 -symmetric ligand in which three thiophene-triazole moieties are positioned around a central mesitylene core. The ligand was characterised by X-ray crystallography and forms multinuclear CuII and CuI complexes identified by mass spectrometry and rationalised by density functional theory (DFT). Upon Cu coordination, CuII -TC-Thio becomes a potent DNA binding and cleaving agent. Mechanistic studies reveal DNA recognition occurs exclusively at the minor groove with subsequent oxidative damage promoted through a superoxide- and peroxide-dependent pathway. Single molecule imaging of DNA isolated from peripheral blood mononuclear cells shows that the complex has comparable activity to the clinical drug temozolomide, causing DNA damage that is recognised by a combination of base excision repair (BER) enzymes.

Self-Assembling Nanoarchitectonics of Twisted Nanofibers of Fluorescent Amphiphiles as Chemo-Resistive Sensor for Methanol Detection.

The inhalation, ingestion, and body absorption of noxious gases lead to severe tissue damage, ophthalmological issues, and neurodegenerative disorders; death may even occur when recognized too late. In particular, methanol gas present in traces can cause blindness, non-reversible organ failure, and even death. Even though ample materials are available for the detection of methanol in other alcoholic analogs at ppm level, their scope is very limited because of the use of either toxic or expensive raw materials or tedious fabrication procedures. In this paper, we report on a simple synthesis of fluorescent amphiphiles achieved using a starting material derived from renewable resources, this material being methyl ricinoleate in good yields. The newly synthesized bio-based amphiphiles were prone to form a gel in a broad range of solvents. The morphology of the gel and the molecular-level interaction involved in the self-assembly process were thoroughly investigated. Rheological studies were carried out to probe the stability, thermal processability, and thixotropic behavior. In order to evaluate the potential application of the self-assembled gel in the field of sensors, we performed sensor measurements. Interestingly, the twisted fibers derived from the molecular assembly could be able to display a stable and selective response towards methanol. We believe that the bottom-up assembled system holds great promise in the environmental, healthcare, medicine, and biological fields.

Obesity impairs cardiolipin-dependent mitophagy and therapeutic intercellular mitochondrial transfer ability of mesenchymal stem cells.

Mesenchymal stem cell (MSC) transplantation alleviates metabolic defects in diseased recipient cells by intercellular mitochondrial transport (IMT). However, the effect of host metabolic conditions on IMT and thereby on the therapeutic efficacy of MSCs has largely remained unexplored. Here we found impaired mitophagy, and reduced IMT in MSCs derived from high-fat diet (HFD)-induced obese mouse (MSC-Ob). MSC-Ob failed to sequester their damaged mitochondria into LC3-dependent autophagosomes due to decrease in mitochondrial cardiolipin content, which we propose as a putative mitophagy receptor for LC3 in MSCs. Functionally, MSC-Ob exhibited diminished potential to rescue mitochondrial dysfunction and cell death in stress-induced airway epithelial cells. Pharmacological modulation of MSCs enhanced cardiolipin-dependent mitophagy and restored their IMT ability to airway epithelial cells. Therapeutically, these modulated MSCs attenuated features of allergic airway inflammation (AAI) in two independent mouse models by restoring healthy IMT. However, unmodulated MSC-Ob failed to do so. Notably, in human (h)MSCs, induced metabolic stress associated impaired cardiolipin-dependent mitophagy was restored upon pharmacological modulation. In summary, we have provided the first comprehensive molecular understanding of impaired mitophagy in obese-derived MSCs and highlight the importance of pharmacological modulation of these cells for therapeutic intervention. A MSCs obtained from (HFD)-induced obese mice (MSC-Ob) show underlying mitochondrial dysfunction with a concomitant decrease in cardiolipin content. These changes prevent LC3-cardiolipin interaction, thereby reducing dysfunctional mitochondria sequestration into LC3-autophagosomes and thus impaired mitophagy. The impaired mitophagy is associated with reduced intercellular mitochondrial transport (IMT) via tunneling nanotubes (TNTs) between MSC-Ob and epithelial cells in co-culture or in vivo. B Pyrroloquinoline quinone (PQQ) modulation in MSC-Ob restores mitochondrial health, cardiolipin content, and thereby sequestration of depolarized mitochondria into the autophagosomes to alleviate impaired mitophagy. Concomitantly, MSC-Ob shows restoration of mitochondrial health upon PQQ treatment (MSC-ObPQQ). During co-culture with epithelial cells or transplantation in vivo into the mice lungs, MSC-ObPQQ restores IMT and prevents epithelial cell death. C Upon transplantation in two independent allergic airway inflammatory mouse models, MSC-Ob failed to rescue the airway inflammation, hyperactivity, metabolic changes in epithelial cells. D PQQ modulated MSCs restored these metabolic defects and restored lung physiology and airway remodeling parameters.

Determining the Functional Oligomeric State of Membrane-Associated Protein Oligomers Forming Membrane Pores on Giant Lipid Vesicles.

Several peripheral membrane proteins are known to form membrane pores through multimerization. In many cases, in biochemical reconstitution experiments, a complex distribution of oligomeric states has been observed that may, in part, be irrelevant to their physiological functions. This phenomenon makes it difficult to identify the functional oligomeric states of membrane lipid interacting proteins, for example, during the formation of transient membrane pores. Using fibroblast growth factor 2 (FGF2) as an example, we present a methodology applicable to giant lipid vesicles by which functional oligomers can be distinguished from nonspecifically aggregated proteins without functionality. Two distinct populations of fibroblast growth factor 2 were identified with (i) dimers to hexamers and (ii) a broad population of higher oligomeric states of membrane-associated FGF2 oligomers significantly distorting the original unfiltered histogram of all detectable oligomeric species of FGF2. The presented statistical approach is relevant for various techniques for characterizing membrane-dependent protein oligomerization.

Sepsis modulates aortic AT1 and P2Y6 receptors to produce vascular hyporeactivity in mice.

PURPOSE: Hyporeactivity to vasopressors leading to multiple organ failure is a serious clinical implication in sepsis. Though the regulatory role of purinoceptors in inflammation is reported, their involvement in sepsis-induced vasoplegia is still unknown. Thus we investigated the effect of sepsis on vascular AT1 and P2Y6 receptors. MATERIALS AND METHODS: Polymicrobial sepsis was induced by cecal ligation and puncture in mice. Vascular reactivity was assessed by organ bath study and aortic mRNA expression of AT1 and P2Y6 was quantified by qRT-PCR. RESULTS: Both angiotensin-II and UDP produced higher contractions in the absence of endothelium as well as following inhibition of nitric oxide synthase. Angiotensin-II mediated aortic contraction was antagonized by losartan (AT1 antagonist), but not by PD123319 (AT2 antagonist) whereas UDP-induced aortic contraction was significantly inhibited by MRS2578 (P2Y6 antagonist). In addition, MRS2578 significantly inhibited the contractile response of Ang-II. Compared to SO mice, angiotensin-II and UDP-induced maximum contraction were found to be significantly attenuated in sepsis. Accordingly, aortic mRNA expression of AT1a receptors was significantly down-regulated while that of P2Y6 receptors was significantly increased in sepsis. 1400 W (a selective iNOS inhibitor) significantly reversed angiotensin-II-induced vascular hyporeactivity in sepsis without affecting UDP-induced hypo-reactivity. CONCLUSION: Sepsis-induced vascular hyporeactivity to angiotensin-II is mediated by enhanced expression of iNOS. Moreover, AT1R-P2Y6 cross talk/heterodimerization could be a novel target for regulating vascular dysfunction in sepsis.

Fabrication of mulberry leaf extract (MLE)- and tasar pupal oil (TPO)-loaded silk fibroin (SF) hydrogels and their antimicrobial properties.

Biocomposites have gained tremendous advantages over synthetic composites due to their biocompatibility, sustainable degradation, and ability to easily combine with other substances. In the present study, we have prepared silk fibroin (SF) hydrogel, mulberry leaf extract (MLE), tasar pupal oil (TPO), and their composites, such as TPO-loaded SF hydrogel and MLE-loaded SF hydrogel, and characterized them by using a phase contrast microscope (PCM), scanning electron microscope (SEM) SEM- EDX, and Fourier transform infrared spectroscopy (FTIR). In addition, 1H-NMR was used for profiling of mulberry leaf extract and GC-MS was used to find tasar pupal oil composition. Further, the disc diffusion method evaluated their antimicrobial activities against S. aureus, E. coli, A. flavus, and A. brassicae. PCM, SEM, and FTIR results validated the conjugation of MLE and SF hydrogel composite; 1H-NMR confirmed the 41 metabolites in MLE, and GC-MS established the composition of tasar pupal oil. Since both composites, such as TPO-loaded SF hydrogel and MLE-loaded SF hydrogel, reduced the S. aureus and E. coli activities at all tested concentrations, the antibacterial results were unambiguous in their conclusion. S. aureus could only be inhibited by SF hydrogel at a high concentration (300 g/ml), despite suppressing E. coli growth at all tested concentrations. At 300 g/ml, MLE demonstrated antibacterial action against S. aureus. Furthermore, at a dosage of 300 g/ml, TPO inhibited both S. aureus and E. coli. Both mulberry leaf extract (at 200 and 300 g/ml) and the MLE-loaded SF hydrogel composite displayed antifungal activity against A. flavus at all tested concentrations (100, 200, and 300 g/ml).