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1.
Environ Res ; 212(Pt A): 113135, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35364041

RESUMO

Baseline assessments of marine microbial studies are very limited around ecologically sensitive areas of the Nuclear Power Plant (NPP) site with respect to their occurrence, distribution, role in adaptation, and their potential remediation process. The distribution and diversity of marine microbes are largely dependent on the physicochemical parameters relating to a specific area, especially spore-producing marine actinobacteria are a source for indigenous bioremediation agents. Marine actinobacterial diversity with conventional and 16 S rRNA gene analysis was done with different pre-treatment conditions and selective media. Totally, 170 different strains are identified in genera level and it belongs to 18 genera with dominant by Streptomyces sp. (75species) followed by Nocardiposis sp, (18species) Rhodococcus sp. (14species). Multiple k-dominance plots simplified the perception of marine actinobacteria according to genera level influence to standard stock. This is the first kind of study in India and the results could act as baseline inventory in terms of microbial diversity around NPP sites. Further, a potential strain of Actinomadura sp. (T5S13) produced 243.7 mg/L of EPS and remediate the Uranium radionuclides. The functional group shifting and adsorption nature were also confirmed by SEM with EDS analysis.


Assuntos
Actinobacteria , Urânio , Actinobacteria/genética , Bactérias/genética , DNA Bacteriano , Centrais Nucleares , Filogenia , RNA Ribossômico 16S/genética
2.
Curr Microbiol ; 78(10): 3620-3633, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34448061

RESUMO

The SARS-CoV-2 virus has spread worldwide to cause a full blown pandemic since 2020. To date, several promising synthetic therapeutics are repurposed and vaccines through different stages of clinical trials were approved and being administered, but still the efficacy of the drugs and vaccines are yet to be decoded. This article highlights the importance of traditional medicinal plants and the phytomolecules derived from them, which possess in vitro antiviral and anti-CoV properties and further explores their potential as inhibitors to molecular targets of SARS-CoV-2 that were evaluated by in silico approaches. Botanicals in traditional medicinal systems have been investigated for anti-SARS-CoV-2 activity through in silico and in vitro studies. However, information linking structure of phytomolecules to their antiviral activity is limited. Most phytomolecules with anti-CoV activity were studied for inhibition of the human ACE2 receptor through which the virus enters host cells, and non-structural proteins 3CLpro and PLpro. Although the proteases are ideal anti-CoV targets, information on plant-based inhibitors for the CoV structural proteins, e.g., spike, envelope, membrane, nucleocapsid required further investigations. In absence of scientific evaluations through in vitro and biocompatibility studies, plant-based antivirals fall short as treatment options. Plant-based anti-SARS-CoV-2 therapeutics can be promising alternatives to their synthetic counterparts as they are economical and bear fewer chances of toxicity, side effects, and viral resistance. Our review could provide a systematic overview of the potential phytomolecules which can be repurposed and subjected to further modes of experimental evaluation to qualify for use in treatment and prophylaxis of SARS-CoV-2 infections.


Assuntos
COVID-19 , Antivirais/farmacologia , Humanos , Pandemias , SARS-CoV-2
3.
Curr Microbiol ; 78(2): 502-512, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33389059

RESUMO

Sutures are widely used materials for closing the surgical wounds, and being an inert material, sutures are often colonized with drug-resistant polymicrobial biofilms. Surgical site infection (SSI) is a hospital-acquired infection caused by bacteria and fungi specifically in the sutured sites. Although most of the currently available sutures possess antibacterial property, their ability to prevent biofilm colonization by polymicrobial communities is underexplored. So, the present study shows that extracted chitosan (EC) from crab shells prevented the adherence of Staphylococcus epidermidis and Candida albicans, the predominant members that exist as mixed species at the site of SSI. In comparison with a commercial chitosan, EC showed profound inhibition of slime formation and mixed species biofilm inhibition. Intriguingly, EC-coated sutures could inhibit the growth of both bacterial and fungal pathogens when comparing with a commercial triclosan-coated suture which was active only against the bacterial pathogen. Scanning electron microscopy results revealed inhibition of C. albicans hyphal formation by the EC-coated sutures that is a crucial virulence factor responsible for tissue invasiveness. Collectively, the results of the present study showed that EC from crab shells (discarded material as a recalcitrant biowaste) could be used as an alternative to combat drug-resistant biofilms which are the prime cause for SSIs.


Assuntos
Quitosana , Microbiota , Preparações Farmacêuticas , Biofilmes , Quitosana/farmacologia , Suturas
4.
J Bacteriol ; 196(23): 4057-70, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25225267

RESUMO

The adaptation of Porphyromonas gingivalis to H2O2-induced stress while inducible is modulated by an unknown OxyR-independent mechanism. Previously, we reported that the PG_2212 gene was highly upregulated in P. gingivalis under conditions of prolonged oxidative stress. Because this gene may have regulatory properties, its function in response to H2O2 was further characterized. PG2212, annotated as a hypothetical protein of unknown function, is a 10.3-kDa protein with a cysteine 2-histidine 2 (Cys2His2) zinc finger domain. The isogenic mutant P. gingivalis FLL366 (ΔPG_2212) showed increased sensitivity to H2O2 and decreased gingipain activity compared to the parent strain. Transcriptome analysis of P. gingivalis FLL366 revealed that approximately 11% of the genome displayed altered expression (130 downregulated genes and 120 upregulated genes) in response to prolonged H2O2-induced stress. The majority of the modulated genes were hypothetical or of unknown function, although some are known to participate in oxidative stress resistance. The promoter region of several of the most highly modulated genes contained conserved motifs. In electrophoretic mobility shift assays, the purified rPG2212 protein did not bind its own promoter region but bound a similar region in several of the genes modulated in the PG_2212-deficient mutant. A metabolome analysis revealed that PG2212 can regulate a number of genes coding for proteins involved in metabolic pathways critical for its survival under the conditions of oxidative stress. Collectively, our data suggest that PG2212 is a transcriptional regulator that plays an important role in oxidative stress resistance and virulence regulation in P. gingivalis.


Assuntos
Regulação Bacteriana da Expressão Gênica , Peróxido de Hidrogênio/toxicidade , Estresse Oxidativo , Porphyromonas gingivalis/efeitos dos fármacos , Porphyromonas gingivalis/fisiologia , Estresse Fisiológico , Fatores de Transcrição/metabolismo , DNA Bacteriano/metabolismo , Ensaio de Desvio de Mobilidade Eletroforética , Deleção de Genes , Perfilação da Expressão Gênica , Porphyromonas gingivalis/genética , Ligação Proteica , Fatores de Transcrição/genética , Dedos de Zinco
5.
J Bacteriol ; 194(6): 1582-92, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22247513

RESUMO

Porphyromonas gingivalis, the causative agent of adult periodontitis, must maintain nitric oxide (NO) homeostasis and surmount nitric oxide stress from host immune responses or other oral bacteria to survive in the periodontal pocket. To determine the involvement of a putative hydroxylamine reductase (PG0893) and a putative nitrite reductase-related protein (PG2213) in P. gingivalis W83 NO stress resistance, genes encoding those proteins were inactivated by allelic exchange mutagenesis. The isogenic mutants P. gingivalis FLL455 (PG0893ermF) and FLL456 (PG2213ermF) were black pigmented and showed growth rates and gingipain and hemolytic activities similar to those of the wild-type strain. P. gingivalis FLL455 was more sensitive to NO than the wild type. Complementation of P. gingivalis FLL455 with the wild-type gene restored the level of NO sensitivity to a level similar to that of the parent strain. P. gingivalis FLL455 and FLL456 showed sensitivity to oxidative stress similar to that of the wild-type strain. DNA microarray analysis showed that PG0893 and PG2213 were upregulated 1.4- and 2-fold, respectively, in cells exposed to NO. In addition, 178 genes were upregulated and 201 genes downregulated more than 2-fold. The majority of these modulated genes were hypothetical or of unknown function. PG1181, predicted to encode a transcriptional regulator, was upregulated 76-fold. Transcriptome in silico analysis of the microarray data showed major metabolomic variations in key pathways. Collectively, these findings indicate that PG0893 and several other genes may play an important role in P. gingivalis NO stress resistance.


Assuntos
Farmacorresistência Bacteriana , Óxido Nítrico/toxicidade , Oxirredutases/metabolismo , Porphyromonas gingivalis/efeitos dos fármacos , Porphyromonas gingivalis/fisiologia , Estresse Fisiológico , Deleção de Genes , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Teste de Complementação Genética , Análise em Microsséries , Oxirredutases/genética , Porphyromonas gingivalis/enzimologia , Porphyromonas gingivalis/genética
6.
Microbiology (Reading) ; 158(Pt 10): 2465-2479, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22745271

RESUMO

Porphyromonas gingivalis, an anaerobic oral pathogen implicated in adult periodontitis, can exist in an environment of oxidative stress. To evaluate its adaptation to this environment, we have assessed the response of P. gingivalis W83 to varying levels and durations of hydrogen peroxide (H(2)O(2))-induced stress. When P. gingivalis was initially exposed to a subinhibitory concentration of H(2)O(2) (0.1 mM), an adaptive response to higher concentrations could be induced. Transcriptome analysis demonstrated that oxidative stress can modulate several functional classes of genes depending on the severity and duration of the exposure. A 10 min exposure to H(2)O(2) revealed increased expression of genes involved in DNA damage and repair, while after 15 min, genes involved in protein fate, protein folding and stabilization were upregulated. Approximately 9 and 2.8% of the P. gingivalis genome displayed altered expression in response to H(2)O(2) exposure at 10 and 15 min, respectively. Substantially more genes were upregulated (109 at 10 min; 47 at 15 min) than downregulated (76 at 10 min; 11 at 15 min) by twofold or higher in response to H(2)O(2) exposure. The majority of these modulated genes were hypothetical or of unknown function. One of those genes (pg1372) with DNA-binding properties that was upregulated during prolonged oxidative stress was inactivated by allelic exchange mutagenesis. The isogenic mutant P. gingivalis FLL363 (pg1372 : : ermF) showed increased sensitivity to H(2)O(2) compared with the parent strain. Collectively, our data indicate the adaptive ability of P. gingivalis to oxidative stress and further underscore the complex nature of its resistance strategy under those conditions.


Assuntos
Adaptação Fisiológica , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica , Peróxido de Hidrogênio/farmacologia , Estresse Oxidativo/fisiologia , Porphyromonas gingivalis/efeitos dos fármacos , Transcriptoma , Proteínas de Bactérias/genética , Humanos , Mutação , Análise de Sequência com Séries de Oligonucleotídeos , Estresse Oxidativo/efeitos dos fármacos , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/crescimento & desenvolvimento , Porphyromonas gingivalis/fisiologia
7.
Int J Biol Macromol ; 194: 870-881, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34843816

RESUMO

Marine ecosystem associated organisms are an affluent source of bioactive compounds. Polysaccharides with unique structural and practical entities have gained special studies interest inside the current biomedical zone. Polysaccharides are the main components of marine algae, plants, animals, insects, and microorganisms. In recent times research on seaweed is more persistent for extraction of natural bioactive "Sulfated polysaccharides" (SPs). The considerable amount of SP exists in the algae in the form of fucans, fucoidans, carrageenans, ulvan, etc. Major function of SPs is to act as a defensive lattice towards the infective organism. All SPs possess the high potential and possess a broad range of therapeutic applications as antitumor, immunomodulatory, vaccine adjuvant, anti-inflammatory, anticoagulant, antiviral, antiprotozoal, antimicrobial, antilipemic, therapy of regenerative medicine, also in drug delivery and tissue engineering application. This review aims to discuss the biomedicine applications of sulfated polysaccharides from marine seaweeds.


Assuntos
Organismos Aquáticos/química , Pesquisa Biomédica , Polissacarídeos/química , Sulfatos/química , Materiais Biocompatíveis/química , Fenômenos Químicos , Carboidratos da Dieta , Sistemas de Liberação de Medicamentos , Desenvolvimento de Medicamentos , Estrutura Molecular , Plantas/química , Polissacarídeos/farmacologia , Alga Marinha/química , Engenharia Tecidual
8.
Sci Rep ; 12(1): 19250, 2022 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-36357447

RESUMO

Recent breakthroughs in the field of nanoparticle-based therapeutic delivery methods have changed the standpoint of cancer therapy by effectively delaying the process of disease development. Nanoparticles have a unique capacity of good penetrating ability than other therapeutic leads used in traditional therapeutics, and also, they have the highest impact on disease management. In the current study isolongifolene-loaded Chitosan nanoparticles have been formulated, synthesized and then characterized by the use of Fourier Transform Infrared Spectroscopy, X-ray Diffraction, Scanning Electron Microscopy and Transmission Electron Microscopy. Further, the characterized chitosan nano formulation was evaluated for hemocompatibility, plasma stability, and in-vitro release. Isolongifolene-loaded chitosan nanoparticles were found to be compatible with plasma and also, they exhibited a constant release pattern. Hence, chitosan-loaded nanoparticles could be employed as an excellent adjuvant in cancer therapeutic, to combat the multi-drug resistance in solid tumors.


Assuntos
Quitosana , Nanopartículas , Neoplasias , Quitosana/química , Nanopartículas/química , Microscopia Eletrônica de Transmissão , Microscopia Eletrônica de Varredura , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X , Tamanho da Partícula , Portadores de Fármacos/química , Neoplasias/tratamento farmacológico
9.
Sci Rep ; 12(1): 4792, 2022 03 21.
Artigo em Inglês | MEDLINE | ID: mdl-35314727

RESUMO

Marine wastes pose a great threat to the ecosystem leading to severe environmental hazards and health issues particularly the shellfish wastes. The shellfish waste which contains half of the amount of chitin can be efficiently transformed into useful products. Various approaches for the hydrolysis of chitin like physical, chemical, and enzymatic processes are there. Still, the use of enzyme chitinase is well documented as an effective and eco-friendly method. The present study summarizes the isolation of chitinase enzyme producing bacteria from different shrimp waste disposal sites in Parangipettai (India), and the possible use of an enzyme hydrolyzate as an immunostimulant to Asian Seabass (Lates calcarifer). The potential chitinase-producing bacteria were identified by 16S rRNA gene sequencing as Stenotrophomonas maltophilia. After purification, the chitinase specific activity was 5.01 (U/ml) and the protein content was 72 mg and the recovery rate was 48.06%. The optimum pH and temperature for the chitinolytic activity were 6.5 and at 35-50 °C, respectively. The animal experiment trial was done with our feed supplements which included 0.0 (control), 0.5%, 1% and 2% of chitin degraded product. All the supplementary feed had an optimal 42% (w/w) of crude protein. The feed protein level was 41-43% on average and gross energy was 13-17 kcal/g and the feed was observed to exhibit a significantly higher (p < 0.05) survival rate, condition factor, specific growth rates, and body weight gain was also found to be promising compared to other fishes fed with control diet only. The red blood cells (RBC) and white blood cell (WBC) counts were found to increase significantly after being challenged with infection in animals fed with chitin derivatives from 1st week to 3rd week when compared to the control. The hematocrit (Hct) values were low on the 2nd and 3rd week in infected fish fed with chitin derivatives. This low level was due to infection lyses of the red blood cells and increased nitro blue tetrazolium reduction. The control diet-fed fish showed 70% mortality but the chitin derivative supplemented fishes showed only 20% mortality post-infection. The results of the study encompass that the use of chitin-derivate enriched feed further is taken into large-scale approaches thereby benefitting the aquaculture sector.


Assuntos
Quitinases , Perciformes , Stenotrophomonas maltophilia , Animais , Quitina/metabolismo , Quitinases/metabolismo , Dieta , Ecossistema , Peixes/metabolismo , Perciformes/metabolismo , RNA Ribossômico 16S/genética , Stenotrophomonas maltophilia/metabolismo
10.
Sci Rep ; 12(1): 3217, 2022 02 25.
Artigo em Inglês | MEDLINE | ID: mdl-35217708

RESUMO

The cultural microbiomes of 27 bacteria colonies were isolated from Mugil cephalus for analysis of the antibacterial and antagonistic activities. A potent probiotic bacterium was characterized using16S r RNA sequencing. The potent strain was added to fish diet to perform the challenge test and to study the growth and immunological parameter. The extracellular proteins from the probiotic were collected and characterized using MALDI TOF/TOF. Out of G27, G9 strain inhibited all the five pathogenic strains. An isolated bacterium was identified as Bacillus subtilis PRBD09 with accession number KF765648. After 35 days of feeding period B. subtilis PRBD09 enhance the both cellular and humoral immune responses, which responsible for survive of the Mugil cephalus against Aeromonas hydrophila infection. The MALDI TOF sample 08 and 09 were recognized as hypothetical proteins based on the MALDI TOF sample. A cytidinedeaminase was found in samples 10, 11, and 12. Extracellular proteins may be involved for the immunological increase in Mugil cephalus against Aeromonas hydrophila, according to the current research.


Assuntos
Doenças dos Peixes , Infecções por Bactérias Gram-Negativas , Probióticos , Smegmamorpha , Aeromonas hydrophila/fisiologia , Animais , Bacillus subtilis , Doenças dos Peixes/microbiologia , Peixes , Infecções por Bactérias Gram-Negativas/microbiologia , Sistema Imunitário/patologia , Imunidade Inata , Probióticos/farmacologia
11.
Infect Immun ; 79(7): 2779-91, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21502589

RESUMO

The Porphyromonas gingivalis recombinant VimA can interact with the gingipains and several other proteins, including a sialidase. Sialylation can be involved in protein maturation; however, its role in virulence regulation in P. gingivalis is unknown. The three sialidase-related proteins in P. gingivalis showed the characteristic sialidase Asp signature motif (SXDXGXTW) and other unique domains. To evaluate the roles of the associated genes, randomly chosen P. gingivalis isogenic mutants created by allelic exchange and designated FLL401 (PG0778::ermF), FLL402 (PG1724::ermF), and FLL403 (PG0352::ermF-ermAM) were characterized. Similar to the wild-type strain, FLL402 and FLL403 displayed a black-pigmented phenotype in contrast to FLL401, which was not black pigmented. Sialidase activity in P. gingivalis FLL401 was reduced by approximately 70% in comparison to those in FLL402 and FLL403, which were reduced by approximately 42% and 5%, respectively. Although there were no changes in the expression of the gingipain genes, their activities were reduced by 60 to 90% in all the isogenic mutants compared to that for the wild type. Immunoreactive bands representing the catalytic domains for RgpA, RgpB, and Kgp were present in FLL402 and FLL403 but were missing in FLL401. While adhesion was decreased, the capacity for invasion of epithelial cells by the isogenic mutants was increased by 11 to 16% over that of the wild-type strain. Isogenic mutants defective in PG0778 and PG0352 were more sensitive to hydrogen peroxide than the wild type. Taken together, these results suggest that the P. gingivalis sialidase activity may be involved in regulating gingipain activity and other virulence factors and may be important in the pathogenesis of this organism.


Assuntos
Metaloendopeptidases/metabolismo , Neuraminidase/metabolismo , Porphyromonas gingivalis/patogenicidade , Fatores de Virulência/genética , Fatores de Virulência/metabolismo , Adesinas Bacterianas/genética , Adesinas Bacterianas/metabolismo , Aderência Bacteriana , Linhagem Celular Tumoral , Cisteína Endopeptidases/genética , Cisteína Endopeptidases/metabolismo , Eletroforese em Gel de Poliacrilamida , Cisteína Endopeptidases Gingipaínas , Células HeLa , Humanos , Immunoblotting , Metaloendopeptidases/genética , Mutação , Neuraminidase/genética , Porphyromonas gingivalis/enzimologia , Porphyromonas gingivalis/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência
12.
J Biomol Struct Dyn ; 39(6): 2106-2117, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32212961

RESUMO

Genes encoding proteins with A20/AN1 zinc-finger domains, belonging to the stress associated protein (SAP) gene family, are present in all eukaryotes and play a decisive role in plant response to diverse physiological and molecular activities particularly on biotic and abiotic stresses (AbS). In this first and foremost study, global transcriptome analysis of members of the SAP gene family was carried out in C3 model-Oryza sativa (OsSAP) aiming at the identification of OsSAP genes activated in response to unique or Combined AbS (CAbS). Based on the available spatio-temporal and phytohormonal RNA-Seq expression profile datasets, nine OsSAP genes were filtered out and identified by a differential expression signature noted in various tissues as well as plant hormones. Comparative genome ideogram of OsSAP genes confirmed the orthologous collinearity with C4 panicoid genomes. Interactome of these genes, revealed the molecular cross-talks of OsSAP. Thus, the computational expression signature of OsSAP genes led to a better understanding of gene dynamism in diverse developmental tissues/organs. Transcriptional regulation analysis of key OsSAP genes in response to stress (drought and salinity) suggested the novel role of OsSAP1, OsSAP2, OsSAP5, OsSAP7, OsSAP8 and OsSAP11 in AbS. Altogether, the study provides deeper insights on molecular characteristics of OsSAP genes, which could be deployed further to decipher their precise functional roles in AbS responses.Communicated by Ramaswamy H. Sarma.


Assuntos
Oryza , Perfilação da Expressão Gênica , Proteínas de Choque Térmico , Oryza/genética , Oryza/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Fisiológico/genética , Transcriptoma
13.
Chemosphere ; 281: 130746, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34022595

RESUMO

Globally, the disposal of shellfishery waste is a major challenge and causes a risk to the coastal region. For potential development in aquaculture, the use of safe supplements to improve fish production and health is important. Chitosan (CS) used as feed additives for several fish species that enhanced production and immunity. The present study was intended to assess the effect of feed additives N-acetyl-d-glucosamine (NAG) loaded chitosan nanoparticles (CSNPs) on productivity, survival rate, and protein conversion efficiency of Oreochromis niloticus (L.). This is the first report on the effect of CSNPs and NAG loaded CSNPs as feed additives enhanced growth performance and non-specific immunity of O. niloticus. CSNPs and NAG loaded CSNPs were synthesized and characterized by scanning and transmission electron microscope, FT-IR, X-ray diffraction, particle size distribution, and zeta sizer. Fish (15.30 ± 0.23 g) administered diets fortified with 0.0, 0.25, 0.5, 1.0, and 2.0 g CSNPs/kg feed loaded with NAG for 45 d. The diets containing 1.0 g/kg NAG loaded CSNPs enhanced specific growth rate, weight gain, survival rate, respiratory burst, and lysozyme activities of tilapia compared control group. The data shows biologically active CSNPs and NAG loaded CSNPs are potent antimicrobial agents against selected bacterial pathogens. In conclusion, the findings suggested that the dietary supplement containing NAG loaded CSNPs significantly increased immune-modulatory properties, growth performance, and enhanced their disease resistance of Nile tilapia.


Assuntos
Quitosana , Ciclídeos , Doenças dos Peixes , Nanopartículas , Ração Animal/análise , Animais , Quitina , Dieta/veterinária , Suplementos Nutricionais/análise , Glucosamina , Espectroscopia de Infravermelho com Transformada de Fourier
14.
Microbiology (Reading) ; 156(Pt 7): 2180-2193, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20378652

RESUMO

The Porphyromonas gingivalis vimA gene has been previously shown to play a significant role in the biogenesis of gingipains. Further, in P. gingivalis FLL92, a vimA-defective mutant, there was increased auto-aggregation, suggesting alteration in membrane surface proteins. In order to determine the role of the VimA protein in cell surface biogenesis, the surface morphology of P. gingivalis FLL92 was further characterized. Transmission electron microscopy demonstrated abundant fimbrial appendages and a less well defined and irregular capsule in FLL92 compared with the wild-type. In addition, atomic force microscopy showed that the wild-type had a smoother surface compared with FLL92. Western blot analysis using anti-FimA antibodies showed a 41 kDa immunoreactive protein band in P. gingivalis FLL92 which was missing in the wild-type P. gingivalis W83 strain. There was increased sensitivity to globomycin and vancomycin in FLL92 compared with the wild-type. Outer membrane fractions from FLL92 had a modified lectin-binding profile. Furthermore, in contrast with the wild-type strain, nine proteins were missing from the outer membrane fraction of FLL92, while 20 proteins present in that fraction from FLL92 were missing in the wild-type strain. Taken together, these results suggest that the VimA protein affects capsular synthesis and fimbrial phenotypic expression, and plays a role in the glycosylation and anchorage of several surface proteins.


Assuntos
Proteínas de Bactérias/metabolismo , Membrana Celular/metabolismo , Porphyromonas gingivalis/metabolismo , Fatores de Virulência/metabolismo , Antibacterianos/farmacologia , Cápsulas Bacterianas/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Membrana Celular/química , Membrana Celular/genética , Fímbrias Bacterianas/metabolismo , Humanos , Conformação Molecular , Mutação , Porphyromonas gingivalis/química , Porphyromonas gingivalis/efeitos dos fármacos , Porphyromonas gingivalis/genética , Fatores de Virulência/química , Fatores de Virulência/genética
15.
J Biotechnol ; 317: 59-69, 2020 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-32353392

RESUMO

Antimicrobial resistance is a major public health concern in infection control. Hence, a multi-pronged approach is necessary to curb the severity of infections. The present study entails the identification of docosanol (fatty alcohol) from Streptomyces as a novel antibiofilm agent which can target the virulence factors of MRSA. Results showed that docosanol as a potent antibiofilm agent and found to inhibit several virulence factors of MRSA. The antibiofilm efficacy of docosanol analyzed through light and scanning electron microscopy showed a significant reduction in adherent cells. Moreover, analysis of three-dimensional structure of biofilm matrix by confocal laser scanning microscope demonstrated effective antibiofilm potential of docosanol. In addition, docosanol reduced the survival rate of MRSA in healthy human blood and enhanced the neutrophil-mediated killing by interfering with hemolysin production. RT-qPCR analysis revealed the down regulation of several virulence genes, possibly by affecting the expression of the accessory gene regulator (agr) system and transcriptional regulator sarA. These findings suggest that docosanol could effectively reduce the biofilm phenotype and virulence production, and thus becomes a promising candidate to treat MRSA infections.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Álcoois Graxos/farmacologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Fatores de Virulência/metabolismo , Animais , Eritrócitos , Hemólise/efeitos dos fármacos , Humanos , Staphylococcus aureus Resistente à Meticilina/metabolismo , Ovinos , Transcriptoma/efeitos dos fármacos
16.
J Photochem Photobiol B ; 203: 111773, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31931385

RESUMO

Glioma is the prime cause of cancer allied mortality in adolescent people and it accounts about 80% of all malignant tumours. Eugenol is a major bioactive constituent present in the essential oils with numerous pharmacological benefits including nueroprotective activity. The major drawback of eugenol is its extreme volatile property and oxygen sensitivity therefore we increased the efficacy of drug; eugenol by encapsulating with chitosan polymer. Eugenol loaded chitosan polymer (EuCs) was characterized using FTIR, XRD, SEM, HR-TEM analysis and the encapsulation, drug release efficacy was assessed at in vitro condition. The induction of autophagy and anticancer efficacy of EuCs on glioma cells was evaluated with rat C6 glioma cells using MTT assay, acridine orange staining, immunocytochemical analysis of NFκß protein expression and FLOW cytometric analysis. The anti-metastatic property of Eu-CS was assessed by immunoblotting and RT-PCR analysis of epithelial mesenchymal transition protein expression in EuCs treated rat C6 glioma cells. Our characterization analysis proves that EuCs possess essential physical and functional properties of copolymer to be utilized as a drug. Further the MTT analysis and AO staining confirms even in the presence of oncogenic inducer and autophagic inhibitors, EuCs exhibits apoptotic potency on rat C6 glioma cells. The result of immunocytochemical studies depicts the inhibition of NFκß protein expression and flow cytometry studies confirm apoptosis induction by EuCs. The inhibition of metastasis by EuCs was proven by the decrease in epithelial mesenchymal transition protein expression in Eu-Cs treated rat C6 glioma cells. Over all our results authentically confirms eugenol loaded chitosan nanopolymer persuasively induces apoptosis and inhibits metastasis in rat C6 glioma cells.


Assuntos
Antineoplásicos/química , Apoptose/efeitos dos fármacos , Quitosana/química , Eugenol/química , Metaloproteinase 9 da Matriz/metabolismo , Nanoestruturas/química , Transdução de Sinais/efeitos dos fármacos , Animais , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Portadores de Fármacos/química , Eugenol/farmacologia , Glioma/metabolismo , Glioma/patologia , NF-kappa B/metabolismo , Ratos , Ativador de Plasminogênio Tipo Uroquinase/metabolismo
17.
Sci Rep ; 10(1): 11898, 2020 07 17.
Artigo em Inglês | MEDLINE | ID: mdl-32681120

RESUMO

Marine pollution is a significant issue in recent decades, with the increase in industries and their waste harming the environment and ecosystems. Notably, the rise in shellfish industries contributes to tons of shellfish waste composed of up to 58% chitin. Chitin, the second most ample polymer next to cellulose, is insoluble and resistant to degradation. It requires chemical-based treatment or enzymatic hydrolysis to cleave the chitin polymers. The chemical-based treatment can lead to environmental pollution, so to solve this problem, enzymatic hydrolysis is the best option. Moreover, the resulting biopolymer by-products can be used to boost the fish immune system and also as drug delivery agents. Many marine microbial strains have chitinase producing ability. Nevertheless, we still lack an economical and highly stable chitinase enzyme for use in the industrial sector. So we isolate a novel marine bacterial strain Achromobacter xylosoxidans from the shrimp waste disposal site using chitin minimal medium. Placket-Burman and central composite design statistical models for culture condition optimisation predicted a 464.2 U/ml of chitinase production. The culture conditions were optimised for maximum chitinase production recording up to 467 U/ml. This chitinase from the A. xylosoxidans was 100% active at an optimum temperature of 45 °C (withstand up to 55 °C) and pH 8 with 80% stability. The HPLC analysis of chitinase degraded shellfish waste reveals a major amino acid profile composition-arginine, lysine, aspartic acid, alanine, threonine and low levels of isoleucine and methionine. These chitinase degraded products and by-products can be used as supplements in the aquaculture industry.


Assuntos
Achromobacter denitrificans/enzimologia , Achromobacter denitrificans/isolamento & purificação , Quitina/metabolismo , Quitinases/biossíntese , Crustáceos/microbiologia , Eliminação de Resíduos , Aminoácidos/análise , Animais , Quitina/química , Quitinases/isolamento & purificação , Estabilidade Enzimática , Concentração de Íons de Hidrogênio , Filogenia , Temperatura
18.
PLoS One ; 14(9): e0223145, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31545847

RESUMO

[This corrects the article DOI: 10.1371/journal.pone.0063367.].

19.
Sci Rep ; 8(1): 9270, 2018 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-29915249

RESUMO

The diversity in plant metabolites with improved phytonutrients is essential to achieve global food security and sustainable crop yield. Our study using computational metabolomics genome wide association study (cmGWAS) reports on a comprehensive profiling of threonine (Thr) metabolite in rice. Sixteen abiotic stress responsive (AbSR) - Thr metabolite producing genes (ThrMPG), modulate metabolite levels and play a significant role determining both physiological and nutritional importance of rice. These AbSR-ThrMPG were computationally analysed for their protein properties using OryzaCyc through plant metabolic network analyser. A total of 1373 and 1028 SNPs were involved in complex traits and genomic variations. Comparative mapping of AbSR-ThrMPG revealed the chromosomal colinearity with C4 grass species. Further, computational expression pattern of these genes predicted a differential expression profiling in diverse developmental tissues. Protein interaction of protein coding gene sequences revealed that the abiotic stresses (AbS) are multigenic in nature. In silico expression of AbSR-ThrMPG determined the putative involvement in response to individual AbS. This is the first comprehensive genome wide study reporting on AbSR -ThrMPG analysis in rice. The results of this study provide a pivotal resource for further functional investigation of these key genes in the vital areas of manipulating AbS signaling in rice improvement.


Assuntos
Adaptação Fisiológica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Oryza/genética , Oryza/fisiologia , Estresse Fisiológico , Treonina/metabolismo , Vias Biossintéticas/genética , Simulação por Computador , Sequência Conservada , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Anotação de Sequência Molecular , Motivos de Nucleotídeos/genética , Oryza/metabolismo , Filogenia , Polimorfismo de Nucleotídeo Único/genética , Fatores de Tempo
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