Nitric oxide promotes cell-matrix adhesion of endothelial progenitor cells under hypoxia condition via ITGA5 CpG promoter demethylation.

Hypoxia or low oxygen tension causes changes in the structure and functional phenotype of the endothelial progenitor cells (EPCs). EPCs are found to be involved in angiogenesis and vascular repair. However, EPC's role in cell-matrix adhesion under hypoxia conditions is not clearly established. Nitric oxide (NO) exerts a wide range of biological functions, especially in regulating the mobilization and vascular repair of EPCs. In contrast, the link between NO and its role in cell-matrix deadhesion under hypoxia is not studied yet. Here, we investigated the protective role of NO in hypoxia-induced cell-matrix deadhesion of EPCs through an epigenetic mechanism. The EPCs were exposed to 2% hypoxia in the presence or absence of 10 µM Spermine NONOate (NO donor). The result demonstrates that hypoxia exposure intensified mitochondrial oxidative damage and energy defects. Using miScript miRNA qPCR array-based screening, the study found miR-148 as a novel target of hypoxia-induced DNMT1 activation. Mechanistically, the study discovered that hypoxia suppressed miR-148 levels and stimulated EPCs cell-matrix deadhesion via increasing DNMT1 mediated Integrin alpha-5 (ITGA5) CpG promoter hypermethylation. Treatment with a mitochondria-targeted antioxidant, MitoTEMPO, or epigenetic DNMT inhibitor, 5'-azacitidine, or miR-148 overexpression in hypoxic EPCs culture, prevented the cell-matrix deadhesion compared to hypoxic EPCs. Further, treatment of spNO or transient expression of eNOS-GFP attenuated hypoxia-induced cell-matrix deadhesion via inhibition of ITGA5 CpG island promoter methylation. In conclusion, the study provides evidence that NO is essential for cell-matrix adhesion of EPCs by epigenetically mitigating ITGA5 CpG promoter hypermethylation under hypoxia conditions. This finding uncovers the previously undefined mechanism of NO-mediated diminution of hypoxia-induced cell-matrix deadhesion and dysfunction induced by low oxygen tension.

miRNA-15/IL-10Rα axis promotes Kabasura Kudineer (Indian traditional Siddha formulation) induced immunomodulation by suppressing oxidative stress.

ETHNOPHARMACOLOGICAL RELEVANCE: Kabasura Kudineer (KK), the traditional Indian medicine of Siddha, effectively manages common respiratory symptoms such as flu, cold, and fever. However, there is no evidence of the immunomodulatory capacity of KK in the cultured Jurkat T-lymphocytes under the LPS insult studied. AIM OF THE STUDY: Assess the effect of the traditional Indian medicine of Siddha, Kabasura Kudineer (KK) on immunomodulation by suppressing oxidative damage in cultured Jurkat T cells in vitro. The miRNA activity on anti-inflammatory gene receptors and cellular nitric oxide levels also was studied. MATERIALS AND METHODS: Jurkat T cells were exposed to LPS treatment in the presence or absence of KK. Cell viability and nitric oxide (NO) were measured with MTT and Griess assay. Cellular antioxidant systems (glutathione and SOD) were determined using glutathione and SOD assay. Lipid peroxidation was measured using an MDA assay. MiRNA-15a-5p expression was performed using microRNA qPCR Assays. Both inflammatory and anti-inflammatory genes (IL-6, IL-1, IL-10, IL-13) were performed using a qPCR and ELISA assay. RESULTS: The data showed that reduced cell proliferation and exaggerated NO production was observed in LPS treated condition compared to the control condition. Further, LPS treatment increased lipid peroxidation and reduced antioxidant enzyme activities (SOD and glutathione) in cultured Jurkat T cells. However, treatment with KK or N-acetyl cysteine (NAC; antioxidant) treatment mitigates the above effect. Mechanistically, LPS-induced oxidative stress upregulated miR- 15-5p expression and suppressed IL-10 Receptor alpha (IL-10Rα) by binding to its 3'-UTR region. The deregulated expression of IL-10Rα expression leads to increased IL-6 and IL-1ß expression in LPS-induced Jurkat T cells; however, treatment with KK or NAC reversed the above effects. CONCLUSION: Collectively, our study revealed the previously undefined mechanistic role of Kabasura Kudineer (KK) that alleviates the LPS-induced oxidative damage associated with inflammation by inhibiting the miRNA-15-5p/IL-10Rα axis.

A case study of flood frequency analysis by intercomparison of graphical linear log-regression method and Gumbel's analytical method in the Vaigai river basin of Tamil Nadu, India.

Flood Frequency Analysis (FFA) is a systematic tool used for an efficient hydraulic structures design, operation and maintenance. An effort is made to study and compare the Linear Log Regression Graphical Method (LLRGM) and Gumbel's Analytical Method (GAM) to assess the future flood magnitude for any given Return Period (RP). Twenty-four years of annual daily peak flood flow value recorded at Vaigai reservoir gauging station between the year 1995 and 2018 was used in the two methods for detailed analysis. The results indicated that the GAM predicts the maximum possible optimum future flood in comparison with the LLRGM. This conclusion was drawn based on the coefficient of determination R2 obtained as 0.8904, which is nearing 1. Based on the analytical method of Gumbel's, the magnitude of frequency factor K has been introduced based on the size of data and coveted RP. The comparative study will provide boon to regulate the storage water to the posterior areas concerning safety and optimum utilization of water for various uses.

Knowledge, attitude, and practice of senior dental students toward management of complications in exodontia.

INTRODUCTION: Dental graduates have a professional responsibility of being competent in managing the complications in exodontia, a very commonly performed procedure. AIM: The aim of this paper was to assess the knowledge, attitude, and practice of senior dental students toward the management of complications in exodontia. MATERIALS AND METHODS: Four hundred dental students participated in the study voluntarily. Verbal consent was obtained after elaborating the purpose of the study. A short validated questionnaire consisting of 12 close-ended multiple choice questions was distributed to all the students. The questionnaires were collected back and results from fully filled questionnaires were tabulated in Microsoft Excel 2007. RESULTS: Nearly 93.4% of the students perform extractions in their practice on a routine basis. Among those 32.5% of the students have encountered complications, out of which only 20% of the students are confident in managing them. The remaining 64.8% of students who have not encountered complications are not confident in managing the same. CONCLUSION: This study revealed the need for increase in cases which will in turn help the students to transfer their theoretical knowledge into professional skills.

In vitro anticancer activity of fucoidan extracted from Sargassum cinereum against Caco-2 cells.

Fucoidan is a marine sulfated polysaccharide, which is extracted from brown seaweed that has a wide range of bioactivities including anti-cancer properties. However, the underlying mechanism of fucoidan on its anti-cancer and apoptotic activity against colon cancer cell line Caco-2 remains to be elucidated. Hence, the present study evaluated the cytotoxicity, apoptotic and anti-cancer activity of fucoidan extracted from brown seaweed Sargassum cinereum against Caco-2 cell line. Cytotoxicity, morphological examination of nuclei, mitochondrial membrane potential, flow cytometry, reactive oxygen species (ROS) formation and detection of apoptotic efficacy of fucoidan were assessed by different assay protocols. Fucoidan inhibited growth of Caco-2 cells in a dose-dependent manner. IC50 concentration of fucoidan was found to be 250 µg/ml. AO/EB, Hoechst and Annexin V/PI staining confirmed the apoptosis induced by fucoidan in Caco-2 cells. Fucoidan was also found to increase ROS production and augment mitochondrial membrane permeability. The findings of the study suggest that fucoidan exerts potent anti-cancer and apoptotic effect on Caco-2 cells by enhancing ROS production. Thus, fucoidan may be used as a promising therapeutic regimen against various cancer cell types.

Acceleration of Intruding Anterior Tooth by Alveolar Corticotomy.

True intrusion is one of the difficult and complex goals to achieve which requires clinical judgment, skills, and experience. Intrusion requires less force level than other tooth movement, but it requires delicacy since the entire stress is concentrated in the apex of the root. Although true intrusion alone is not challenging, eliminating the adverse effect while intruding requires tedious skills of the clinician. A 17-year-old male patient with Class I malocclusion with open bite has a prominent maxillary cortical bone. He had a convex profile with an unpleasant smile. To reduce the bulkiness of cortical bone and to intrude the upper anterior, so the decision was made to perform corticotomy.

Not all the infected develop the disease - A "Lotus and Cactus" model.

The immunogenetic dictum "not all the infected develop the disease" can best be explained by a "Lotus and Cactus" model. Lotuses grow in ponds and cacti in deserts: analogously, we can say that tubercle patient's lung (genetic makeup) functions as an ideal 'broth' for Mycobacterium tuberculosis (M.tb) germs to grow, but not the lungs of an endemic control. HLA association studies from Europe to Asia since 1983 till date, have shown a persistent HLA DR2 (15) association. Further, HLA DR2 and non-DR2 endemic controls showed disparate patterns of immune responses and gene expressions. The host and pathogen MHC diversities, Th1-Th2 paradigm and cytokine circuits all may play a crucial role in TB susceptibility. It is possible to decipher the protective immunity by controlling the known confounders - epidemiological, demographic, socio-biological and also host and pathogen diversities. This has become significant with our understanding on the 'out of Africa' migration and neolithic co-dispersal of M.tb with modern human. Divergence and expansion of various MHCs (eg HLA-DRB1*15, HLA-B*57) and non-MHC alleles in various continents might be responsible for the skewed transmission and distribution of the infectious diseases around the globe. The 'Lotus and Cactus' model proposed here exemplifies this. A holistic genetic epidemiology approach employing modern tools is the need of the hour to better understand infectious disease susceptibility.

Inhibition of quorum-sensing-dependent virulence factors and biofilm formation of clinical and environmental Pseudomonas aeruginosa strains by ZnO nanoparticles.

UNLABELLED: Quorum quenching decreases Pseudomonas aeruginosa virulence factors and biofilm formation, alleviating infections in animal models. Nevertheless, it is usually performed in laboratory strains such as PAO1 and PA14, and studies involving clinical or environmental isolates are scarce. In this work, the effects of ZnO nanoparticles, a potent quorum and virulence quencher for the PAO1 strain, were tested in six clinical strains from cystic fibrosis patients, a furanone C-30 resistant clinical strain from urine, two PA14 gallium resistant mutants, a PA14 C-30 resistant mutant and four environmental isolates. ZnO nanoparticles effectively decreased elastase, pyocyanin, and biofilm formation for most of the strains; regardless their origin or their resistance against the canonical quorum quencher C-30 or the novel antimicrobial gallium. The data indicate ZnO nanoparticles may have a broad spectrum for the quorum quenching of relevant strains and that may be an alternative to treat Ps. aeruginosa recalcitrant infections. SIGNIFICANCE AND IMPACT OF THE STUDY: Virulence inhibition by quorum quenchers in Pseudomonas aeruginosa is usually tested in laboratory strains and studies of their effects in relevant clinical and environmental strains are scarce. This study is significant as the effects of ZnO nanoparticles in QS-dependent virulence factor production were tested in six clinical strains from cystic fibrosis patients, a C-30 resistant clinical strain from urine, two PA14 gallium resistant mutants, a PA14 C-30 resistant mutant, and four environmental isolates. ZnO nanoparticles decreased elastase, pyocyanin, and biofilms for most of the strains; indicating they have broad spectrum and may be an alternative to treat Ps. aeruginosa infections.

Interactions between water temperature and contaminant toxicity to freshwater fish.

Warming of freshwaters as a result of climate change is expected to have complex interactions with the toxicity of contaminants to aquatic organisms. The present study evaluated the effects of temperature on the acute toxicity of endosulfan, chlorpyrifos, and phenol to 3 warm water species of fish-silver perch, rainbowfish, and western carp gudgeon-and 1 cold water species, rainbow trout. Endosulfan was more toxic to silver perch at 30 °C and 35 °C than at 15 °C, 20 °C and 25 °C during short exposures of 24 h, but at 96 h, temperature had no effect on toxicity. Toxicity to rainbow trout increased with increasing temperature, whereas warm water species exhibited maximum toxicity at around 30 °C, decreasing again toward 35 °C. Chlorpyrifos became more toxic to all species with increasing temperature. Phenol toxicity to all species decreased at low to intermediate temperatures; but as temperatures increased further toward the upper thermal limit, phenol became more toxic. Increasing toxicity in the upper thermal range of cold water species may contribute to upstream range contraction in rivers with high toxicant loads. In contrast, warm water species may not exhibit a range shift within rivers as a result of interactions between temperature and toxicity. Catchment management to offset global warming at local scales may present opportunities to mitigate increased toxicity of contaminants to fish.

Complex composite odontoma with characteristic histology.

Odontomas are the most commonly occurring odontogenic tumors, which are considered by many to be hamartomas rather than neoplasms. These clinically asymptomatic tumors are classified into complex and compound odontomas. They are usually discovered in radiographs and rarely cause bony expansion or infection. This paper discusses a case report of a complex odontoma exhibiting all the structural features and defects of enamel, dentine, and cementum in succession, with an overview on its etiology.

Idiopathic multiple impacted unerupted teeth: Case report and discussion.

Multiple impacted permanent teeth are usually related to syndromes, metabolic and hormonal disorders. However, in some cases, impaction of multiple teeth is not associated with any syndrome. In this report, we present a case of 17-year-old male patient with missing teeth. Radiographs revealed multiple impacted permanent teeth, though medical and family history along with physical examination was not suggestive of any syndromes. If other investigations are negative, an idiopathic case of multiple impacted teeth is suggested to be the possible diagnosis. The objective of this report is to increase awareness of such cases especially in the absence of hereditary/genetic/metabolic factors usually inherent in such scenarios. The patient management in such cases needs to be planned specifically from a multidisciplinary standpoint.

Novel pharmacological targets from Indian cone snails.

The oceans are a source of combinatorial library of unique natural products, 'not found in the terrestrial environment'. Marine invertebrates such as sponges, molluscs, bryozoans, tunicates (Urochordata) and their associated microorganisms are the major representatives of promising bioactive compounds. Among these, the predatory molluscan cone snails have evolved with highly structured small and complex array of peptides (more than 50,000) linked to their prey capture and defence. These peptides have become a valuable source of neuro pharmacological targets as many of them selectively modulate ion channels and transporters. A group of scientists from United States, Europe, Australia, Israel and China have been characterized drugs for neuropathic pain and pharmacological targets from the peptides of a few cone snail species. Several are now in Clinical and preclinical development. Less than 1% of the cono peptides are pharmacologically characterized. India has a diversity of 20-30% of total cone snail species distributed worldwide. A group of Indian Scientists have made promising drug discovery programs from Conus peptides. This review will focus on the Conus peptides from Indian cone snails species, their neuro pharmacological targets and future directions.

Two-year impacts on employment and income among adults receiving antiretroviral therapy in Tamil Nadu, India: a cohort study.

OBJECTIVES: to estimate the economic impact of antiretroviral therapy (ART) on employment and income of treated patients as well as a comparison group of pre-ART patients who receive care and support in Tamil Nadu, India. METHODS: a cohort of 1238 HIV-infected patients was followed between 2005 and 2007. Socioeconomic data were collected at 6-month intervals. A total of 515 patients initiated ART during the study period, whereas a comparison group of 723 patients were pre-ART. The impact of ART on four employment outcomes was analyzed: participation in economic activities in the past week, number of hours worked in the past week, individual income earned in the past 30 days and 6 months. Regression models including patient fixed effects were estimated. Data from the comparison group of patients were used to adjust for time trends in employment outcomes. RESULTS: At 6 months after initiation of ART, patients were 10 percentage points more likely to be economically active (P < 0.01) and worked 5.5 additional hours per week (P < 0.01). These increases were over and above those experienced by the comparison group. The estimated 24-month impacts represent a doubling of patients' employment levels at baseline. At 24 months after ART initiation, employment increases remained large and significant. Effects were almost twice as large for men compared with women. Income earned in the past 30 days and 6 months also rose significantly. CONCLUSION: ART resulted in a rapid and sustained increase in employment and income for patients. The results demonstrate that ART can improve the economic outcomes of HIV-infected patients.

Leprosy and the adaptation of human toll-like receptor 1.

Leprosy is an infectious disease caused by the obligate intracellular pathogen Mycobacterium leprae and remains endemic in many parts of the world. Despite several major studies on susceptibility to leprosy, few genomic loci have been replicated independently. We have conducted an association analysis of more than 1,500 individuals from different case-control and family studies, and observed consistent associations between genetic variants in both TLR1 and the HLA-DRB1/DQA1 regions with susceptibility to leprosy (TLR1 I602S, case-control P = 5.7 x 10(-8), OR = 0.31, 95% CI = 0.20-0.48, and HLA-DQA1 rs1071630, case-control P = 4.9 x 10(-14), OR = 0.43, 95% CI = 0.35-0.54). The effect sizes of these associations suggest that TLR1 and HLA-DRB1/DQA1 are major susceptibility genes in susceptibility to leprosy. Further population differentiation analysis shows that the TLR1 locus is extremely differentiated. The protective dysfunctional 602S allele is rare in Africa but expands to become the dominant allele among individuals of European descent. This supports the hypothesis that this locus may be under selection from mycobacteria or other pathogens that are recognized by TLR1 and its co-receptors. These observations provide insight into the long standing host-pathogen relationship between human and mycobacteria and highlight the key role of the TLR pathway in infectious diseases.

Meiotic recombination generates rich diversity in NK cell receptor genes, alleles, and haplotypes.

Natural killer (NK) cells contribute to the essential functions of innate immunity and reproduction. Various genes encode NK cell receptors that recognize the major histocompatibility complex (MHC) Class I molecules expressed by other cells. For primate NK cells, the killer-cell immunoglobulin-like receptors (KIR) are a variable and rapidly evolving family of MHC Class I receptors. Studied here is KIR3DL1/S1, which encodes receptors for highly polymorphic human HLA-A and -B and comprises three ancient allelic lineages that have been preserved by balancing selection throughout human evolution. While the 3DS1 lineage of activating receptors has been conserved, the two 3DL1 lineages of inhibitory receptors were diversified through inter-lineage recombination with each other and with 3DS1. Prominent targets for recombination were D0-domain polymorphisms, which modulate enhancer function, and dimorphism at position 283 in the D2 domain, which influences inhibitory function. In African populations, unequal crossing over between the 3DL1 and 3DL2 genes produced a deleted KIR haplotype in which the telomeric "half" was reduced to a single fusion gene with functional properties distinct from its 3DL1 and 3DL2 parents. Conversely, in Eurasian populations, duplication of the KIR3DL1/S1 locus by unequal crossing over has enabled individuals to carry and express alleles of all three KIR3DL1/S1 lineages. These results demonstrate how meiotic recombination combines with an ancient, preserved diversity to create new KIR phenotypes upon which natural selection acts. A consequence of such recombination is to blur the distinction between alleles and loci in the rapidly evolving human KIR gene family.

Effects of temperature on ventilatory behavior of fish exposed to sublethal concentrations of endosulfan and chlorpyrifos.

The ventilation amplitude and frequency of silver perch Bidyanus bidyanus, and the ventilation frequency of rainbow fish Melanotaenia duboulayi and rainbow trout Oncorhynchus mykiss, were determined at different temperatures upon exposure to endosulfan and chlorpyrifos, respectively. Silver perch and rainbow fish were tested at 15, 20, 25, 30, and 35 degrees C, while rainbow trout was tested at 10, 15, 20, and 25 degrees C. Although some trend of increasing amplitudes with increasing temperature was evident; there was no significant temperature response of ventilation frequency rates over time in silver perch pre-exposed to 10 microg L(-1) endosulfan for 18 h. The rate of ventilation frequency of rainbow fish pre-exposed to 200 microg L(-1) of chlorpyrifos for 96 h was lower in treatments than in the control at 15 degrees C. However, between 20 and 35 degrees C, rates were significantly higher in the treatments than those of the control. In rainbow trout pre-exposed to 100 microg L(-1) of chlorpyrifos, the rates of frequency were significantly lower than those of controls in temperatures between 10 and 20 degrees C but higher at 25 degrees C. The amplitude of silver perch seemed to increase with the increase in temperature; however, the corresponding temperature quotient values at various temperature regimes and over exposure time showed no significant differences. The ventilation frequency of rainbow fish and rainbow trout significantly increased at the higher test temperatures, and their corresponding temperature quotient values for both fish also increased at the elevated temperatures.

Distinct diversity of KIR genes in three southern Indian populations: comparison with world populations revealed a link between KIR gene content and pre-historic human migrations.

By interacting with polymorphic HLA class I molecules, the killer cell immunoglobulin-like receptors (KIR) influence the innate and adaptive immune response to infection. The KIR family varies in gene content and sequence polymorphism, thereby, distinguishing individuals and populations. To investigate KIR diversity in the earliest settlers of India, we have characterized the KIR gene content in three Dravidian-speaking populations (Mollukurumba, Kanikar, and Paravar) from the state of Tamil Nadu, southern India. The activating KIR genes and putative group-B KIR haplotypes were frequent in Paravar and Kanikar, a scenario analogous to those seen previously in other populations of Indian origin, indicating that predominance of group-B KIR haplotypes is the characteristic feature of Indian populations. In contrast, the KIR gene profile of Mollukurumba was more related to Caucasian type. It is not clear whether a local-specific selection or a recent admixture from Iran is responsible for such discrete profile in Mollukurumba. Each southern Indian population had distinct KIR genotype profile. Comparative analyses with world populations revealed that group-B KIR haplotypes were frequent in the natives of India, Australia, and America, the populations associated with those involved in extensive prehistoric human migrations. Whether or not natural selection has acted to enrich group-B KIR haplotypes in these migratory descendants is an issue that requires objective testing.

Unusual selection on the KIR3DL1/S1 natural killer cell receptor in Africans.

Interactions of killer cell immunoglobulin-like receptors (KIRs) with major histocompatibility complex (MHC) class I ligands diversify natural killer cell responses to infection. By analyzing sequence variation in diverse human populations, we show that the KIR3DL1/S1 locus encodes two lineages of polymorphic inhibitory KIR3DL1 allotypes that recognize Bw4 epitopes of protein">HLA-A and HLA-B and one lineage of conserved activating KIR3DS1 allotypes, also implicated in Bw4 recognition. Balancing selection has maintained these three lineages for over 3 million years. Variation was selected at D1 and D2 domain residues that contact HLA class I and at two sites on D0, the domain that enhances the binding of KIR3D to HLA class I. HLA-B variants that gained Bw4 through interallelic microconversion are also products of selection. A worldwide comparison uncovers unusual KIR3DL1/S1 evolution in modern sub-Saharan Africans. Balancing selection is weak and confined to D0, KIR3DS1 is rare and KIR3DL1 allotypes with similar binding sites predominate. Natural killer cells express the dominant KIR3DL1 at a high frequency and with high surface density, providing strong responses to cells perturbed in Bw4 expression.

Fouling deterrent chemical defence in three muricid gastropod egg masses from the Southeast coast of India.

The egg masses of the marine muricid gastropod molluscs Chicoreus virgineus, Chicoreus ramosus and Rapana rapiformis were studied for antifouling activities. The minimum inhibitory concentrations of crude extracts for the inhibition of byssal production and attachment of the brown mussel Perna indica were 650 mug ml(-1), 1150 mug ml(-1) and 925 mug ml(-1) from the three muricid gastropods, respectively. Higher LC(50) values than EC(50) values and 100% recovery of the mussels in the toxicity assay indicated the non-toxic nature of the extracts. The gradient partitioning of the egg mass extracts and subsequent antimicrofouling screening against 40 biofilm bacteria showed wide-spectrum antibacterial activity of the medium polar fraction from C. virgineus; the non-polar fraction from R. rapiformis and both non-polar and medium polar fractions from C. ramosus. The antimicrofouling activity from extracts of the three egg masses was found to be more prominent than antimacrofouling activity. This may be attributed to the targeting of a defence strategy against microbes in order to protect the developing mollusc embryos.