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1.
Brief Bioinform ; 22(2): 1076-1084, 2021 03 22.
Artículo en Inglés | MEDLINE | ID: mdl-33480398

RESUMEN

Viruses are responsible for causing various epidemics and pandemics with a high mortality rate e.g. ongoing SARS-CoronaVirus-2 crisis. The discovery of novel antivirals remains a challenge but drug repurposing is emerging as a potential solution to develop antivirals in a cost-effective manner. In this regard, we collated the information of repurposed drugs tested for antiviral activity from literature and presented it in the form of a user-friendly web server named 'DrugRepV'. The database contains 8485 entries (3448 unique) with biological, chemical, clinical and structural information of 23 viruses responsible to cause epidemics/pandemics. The database harbors browse and search options to explore the repurposed drug entries. The data can be explored by some important fields like drugs, viruses, drug targets, clinical trials, assays, etc. For summarizing the data, we provide overall statistics of the repurposed candidates. To make the database more informative, it is hyperlinked to various external repositories like DrugBank, PubChem, NCBI-Taxonomy, Clinicaltrials.gov, World Health Organization and many more. 'DrugRepV' database (https://bioinfo.imtech.res.in/manojk/drugrepv/) would be highly useful to the research community working to develop antivirals.


Asunto(s)
Antivirales/farmacología , Reposicionamiento de Medicamentos , Pandemias , COVID-19/virología , Bases de Datos Factuales , Humanos , SARS-CoV-2/efectos de los fármacos
2.
BMC Infect Dis ; 20(1): 671, 2020 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-32933490

RESUMEN

BACKGROUND: The increasing antimalarial drug resistance is a significant hindrance to malaria control and elimination programs. For the last six decades, chloroquine (CQ) plus pyrimethamine remains the first-line treatment for P. vivax malaria. Regions where both P. falciparum and P. vivax co-exist, P. vivax is exposed to antifolate drugs due to either misdiagnosis or improper treatment that causes selective drug pressure to evolve. Therefore, the present study aims to estimate antimalarial drug resistance among the complicated and uncomplicated P. vivax patients. METHODS: A total of 143 P. vivax malaria positive patients were enrolled in this study, and DNA was isolated from their blood samples. Pvcrt-o, Pvmdr-1, Pvdhps, and Pvdhfr genes were PCRs amplified, and drug resistance-associated gene mutations were analyzed. Statistical analysis of the drug resistance genes and population diversity was performed using MEGA vs. 7.0.21 and DnaSP v software. RESULTS: Among the CQ resistance marker gene Pvcrt-o, the prevalence of K10 insertion was 17.5% (7/40) and 9.5% (7/73) of complicated and uncomplicated P vivax group isolates respectively. In Pvmdr-1, double mutant haplotype (M958/L1076) was found in 99% of the clinical isolates. Among the pyrimethamine resistance-associated gene Pvdhfr, the double mutant haplotype I13P33F57R58T61N117I173 was detected in 23% (11/48) in complicated and 20% (17/85) in uncomplicated group isolates. In the sulphadoxine resistance-associated Pvdhps gene, limited polymorphism was observed with the presence of a single mutant (D459A) among 16 and 5% of the clinical isolates in the complicated and uncomplicated group respectively. CONCLUSION: The study presents the situations of polymorphism in the antimalarial drug resistance-associated genes and emphasizes the need for regular surveillance. It is imperative for the development of suitable antimalarial drug policy in India.


Asunto(s)
Antimaláricos/uso terapéutico , Resistencia a Medicamentos/genética , Malaria Vivax/tratamiento farmacológico , Plasmodium vivax/genética , Proteínas Protozoarias/genética , Adolescente , Niño , Preescolar , Cloroquina/uso terapéutico , ADN Protozoario/genética , ADN Protozoario/metabolismo , Femenino , Antagonistas del Ácido Fólico/uso terapéutico , Haplotipos , Humanos , India , Masculino , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/genética , Plasmodium vivax/aislamiento & purificación , Polimorfismo de Nucleótido Simple , Adulto Joven
3.
Malar J ; 18(1): 308, 2019 Sep 06.
Artículo en Inglés | MEDLINE | ID: mdl-31492135

RESUMEN

BACKGROUND: Malaria is one of the important vector-borne diseases with high fatality rates in tropical countries. The pattern of emergence and spread of novel antigenic variants, leading to escape of vaccine-induced immunity might be factors responsible for severe malaria. A high level of polymorphism has been reported among malarial antigens which are under selection pressure imposed by host immunity. There are limited reports available on comparative stage-specific genetic diversity among Plasmodium vivax candidate genes in complicated vivax malaria. The present study was planned to study genetic diversity (Pvcsp and Pvs25) among complicated and uncomplicated P. vivax isolates. METHODS: Pvcsp and Pvs2-specific PCRs and DNA sequencing were performed on P. vivax PCR positive samples. Genetic diversity was analysed using appropriate software. RESULTS: The present study was carried out on 143 P. vivax clinical isolates, collected from Postgraduate Institute of Medical Education and Research, Chandigarh. Among the classic and variant types of Pvcsp, the VK210 (99%; 115/116) was found to be predominant in both complicated and uncomplicated group isolates. Out of the various peptide repeat motifs (PRMs) observed, GDRADGQPA (PRM1) and GDRAAGQPA (PRM2) was the most widely distributed among the P. vivax isolates. Whereas among the Pvs25 isolates, 100% of double mutants (E97Q/I130T) in both the complicated (45/45) as well as in the uncomplicated (81/81) group was observed. CONCLUSION: An analysis of genetic variability enables an understanding of the role of genetic variants in severe vivax malaria.


Asunto(s)
Antígenos de Protozoos/genética , Antígenos de Superficie/genética , Variación Genética , Vacunas contra la Malaria/genética , Plasmodium vivax/genética , Proteínas Protozoarias/genética , Adolescente , Adulto , Niño , Femenino , Humanos , India , Masculino , Adulto Joven
4.
J Transl Med ; 16(1): 272, 2018 10 04.
Artículo en Inglés | MEDLINE | ID: mdl-30286756

RESUMEN

BACKGROUND: In the recent years Plasmodium vivax has been reported to cause severe infections associated with mortality. Clinical evaluation has limited accuracy for the early identification of the patients progressing towards the fatal condition. Researchers have tried to identify the serum and the plasma-based indicators of the severe malaria. Discovery of MicroRNA (miRNA) has opened up an era of identification of early biomarkers for various infectious and non-infectious diseases. MicroRNAs (miRNA) are the small non-coding RNA molecules of length 19-24 nts and are responsible for the regulation of the majority of human gene expressions at post transcriptional level. METHODS: We identified the differentially expressed miRNAs by microarray and validated the selected miRNAs by qRT-PCR. We assessed the diagnostic potential of these up-regulated miRNAs for complicated P. vivax malaria. Futher, the bioinformtic analysis was performed to construct protein-protein and mRNA-miRNA networks to identify highly regulated miRNA. RESULTS: In the present study, utility of miRNA as potential biomarker of complicated P. vivax malaria was explored. A total of 276 miRNAs were found to be differentially expressed by miRNA microarray and out of which 5 miRNAs (hsa-miR-7977, hsa-miR-28-3p, hsa-miR-378-5p, hsa-miR-194-5p and hsa-miR-3667-5p) were found to be significantly up-regulated in complicated P. vivax malaria patients using qRT-PCR. The diagnostic potential of these 5 miRNAs were found to be significant with sensitivity and specificity of 60-71% and 69-81% respectively and area under curve (AUC) of 0.7 (p < 0.05). Moreover, in silico analysis of the common targets of up-regulated miRNAs revealed UBA52 and hsa-miR-7977 as majorly regulated hubs in the PPI and mRNA-miRNA networks, suggesting their putative role in complicated P. vivax malaria. CONCLUSION: miR-7977 might act as a potential biomarker for differentiating complicated P. vivax malaria from uncomplicated type. The elevated levels of miR-7977 may have a role to play in the disease pathology through UBA52 or TGF-beta signalling pathway.


Asunto(s)
Biomarcadores/sangre , Malaria Vivax/sangre , Malaria Vivax/diagnóstico , Tamizaje Masivo , Plasmodium vivax/fisiología , Adolescente , Adulto , Algoritmos , Niño , Femenino , Ontología de Genes , Genes Esenciales , Humanos , Malaria Vivax/genética , Masculino , MicroARNs/sangre , MicroARNs/genética , MicroARNs/metabolismo , Mapas de Interacción de Proteínas/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Curva ROC , Reacción en Cadena en Tiempo Real de la Polimerasa , Reproducibilidad de los Resultados , Regulación hacia Arriba/genética , Adulto Joven
5.
J Med Virol ; 87(2): 344-9, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25156964

RESUMEN

Rubella virus outbreaks usually occur when a large numbers of susceptible individuals accumulate. The disease presents clinically with fever and maculopapular rash. The present study reports the investigation of rubella outbreak in a modern and well-planned village near Chandigarh, North India. The blood samples were collected from 39 cases with febrile rash and from 15 age and sex matched healthy controls residing in the same locality and subjected for the detection of Rubella IgM and IgG antibodies by Enzyme linked immunosorbent assay. The throat swabs, urine and blood samples from acute cases were also collected and subjected to RT-PCR using the primers targeting the E1 region. The genetic characterization of the rubella virus was carried out to identify the circulating genotypes. In the present outbreak, 13 laboratory confirmed cases were reported. Rubella IgM antibodies were detected in 12/39 (30.7%) patients. Rubella RNA could be detected in 83.3% (5/6) of urine, 22.2% (2/9) of throat swabs, and 8.3% (1/12) of blood samples. The rubella genotype responsible for the present outbreak was identified as genotype 1a. This outbreak highlights the need for the introduction of rubella vaccine in the National Immunization Programme of India to prevent outbreaks and to aim towards the eradication of this disease. This study reports the presence of genotype 1a in North India for the first time and stresses the need for further molecular work to identify the circulating strains of the virus.


Asunto(s)
Brotes de Enfermedades , Rubéola (Sarampión Alemán)/epidemiología , Adolescente , Adulto , Anticuerpos Antivirales/sangre , Sangre/virología , Preescolar , Ensayo de Inmunoadsorción Enzimática , Femenino , Genotipo , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , India/epidemiología , Lactante , Masculino , Faringe/virología , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Orina/virología , Proteínas del Envoltorio Viral/genética , Adulto Joven
6.
Aging (Albany NY) ; 15(16): 7866-7908, 2023 08 21.
Artículo en Inglés | MEDLINE | ID: mdl-37606991

RESUMEN

Dectin-1 is an innate immune receptor that recognizes and binds ß-1, 3/1, 6 glucans on fungi. We evaluated Dectin-1 function in myeloid cells in a cohort of HIV-positive and HIV-negative young and older adults. Stimulation of monocytes with ß-D-glucans induced a pro-inflammatory phenotype in monocytes of HIV-infected individuals that was characterized by increased levels of IL-12, TNF-α, and IL-6, with some age-associated cytokine increases also noted. Dendritic cells showed a striking HIV-associated increase in IFN-α production. These increases in cytokine production paralleled increases in Dectin-1 surface expression in both monocytes and dendritic cells that were noted with both HIV and aging. Differential gene expression analysis showed that HIV-positive older adults had a distinct gene signature compared to other cohorts characterized by a robust TNF-α and coagulation response (increased at baseline), a persistent IFN-α and IFN-γ response, and an activated dendritic cell signature/M1 macrophage signature upon Dectin-1 stimulation. Dectin-1 stimulation induced a strong upregulation of MTORC1 signaling in all cohorts, although increased in the HIV-Older cohort (stimulation and baseline). Overall, our study demonstrates that the HIV Aging population has a distinct immune signature in response to Dectin-1 stimulation. This signature may contribute to the pro-inflammatory environment that is associated with HIV and aging.


Asunto(s)
Infecciones por VIH , Factor de Necrosis Tumoral alfa , Humanos , Citocinas , Glucanos
7.
J Heart Lung Transplant ; 42(1): 53-63, 2023 01.
Artículo en Inglés | MEDLINE | ID: mdl-37014805

RESUMEN

BACKGROUND: Long term outcomes of lung transplantation are impacted by the occurrence of chronic lung allograft dysfunction (CLAD). Recent evidence suggests a role for the lung microbiome in the occurrence of CLAD, but the exact mechanisms are not well defined. We hypothesize that the lung microbiome inhibits epithelial autophagic clearance of pro-fibrotic proteins in an IL-33 dependent manner, thereby augmenting fibrogenesis and risk for CLAD. METHODS: Autopsy derived CLAD and non-CLAD lungs were collected. IL-33, P62 and LC3 immunofluorescence was performed and assessed using confocal microscopy. Pseudomonas aeruginosa (PsA), Streptococcus Pneumoniae (SP), Prevotella Melaninogenica (PM), recombinant IL-33 or PsA-lipopolysaccharide was co-cultured with primary human bronchial epithelial cells (PBEC) and lung fibroblasts in the presence or absence of IL-33 blockade. Western blot analysis and quantitative reverse transcription (qRT) PCR was performed to evaluate IL-33 expression, autophagy, cytokines and fibroblast differentiation markers. These experiments were repeated after siRNA silencing and upregulation (plasmid vector) of Beclin-1. RESULTS: Human CLAD lungs demonstrated markedly increased expression of IL-33 and reduced basal autophagy compared to non-CLAD lungs. Exposure of co-cultured PBECs to PsA, SP induced IL-33, and inhibited PBEC autophagy, while PM elicited no significant response. Further, PsA exposure increased myofibroblast differentiation and collagen formation. IL-33 blockade in these co-cultures recovered Beclin-1, cellular autophagy and attenuated myofibroblast activation in a Beclin-1 dependent manner. CONCLUSION: CLAD is associated with increased airway IL-33 expression and reduced basal autophagy. PsA induces a fibrogenic response by inhibiting airway epithelial autophagy in an IL-33 dependent manner.


Asunto(s)
Artritis Psoriásica , Pseudomonas , Humanos , Beclina-1/metabolismo , Interleucina-33/metabolismo , Artritis Psoriásica/metabolismo , Pulmón/metabolismo , Autofagia/fisiología
8.
J Heart Lung Transplant ; 40(6): 447-457, 2021 06.
Artículo en Inglés | MEDLINE | ID: mdl-33781665

RESUMEN

BACKGROUND: Recent evidence suggests a role for lung microbiome in occurrence of chronic lung allograft dysfunction (CLAD). However, the mechanisms linking the microbiome to CLAD are poorly delineated. We investigated a possible mechanism involved in microbial modulation of mucosal response leading to CLAD with the hypothesis that a Proteobacteria dominant lung microbiome would inhibit N-myc-interactor (NMI) expression and induce epithelial to mesenchymal transition (EMT). METHODS: Explant CLAD, non-CLAD, and healthy nontransplant lung tissue were collected, as well as bronchoalveolar lavage from 14 CLAD and matched non-CLAD subjects, which were followed by 16S rRNA amplicon sequencing and quantitative polymerase chain reaction (PCR) analysis. Pseudomonas aeruginosa (PsA) or PsA-lipopolysaccharide was cocultured with primary human bronchial epithelial cells (PBEC). Western blot analysis and quantitative reverse transcription (qRT) PCR was performed to evaluate NMI expression and EMT in explants and in PsA-exposed PBECs. These experiments were repeated after siRNA silencing and upregulation (plasmid vector) of EMT regulator NMI. RESULTS: 16S rRNA amplicon analyses revealed that CLAD patients have a higher abundance of phyla Proteobacteria and reduced abundance of the phyla Bacteroidetes. At the genera level, CLAD subjects had an increased abundance of genera Pseudomonas and reduced Prevotella. Human CLAD airway cells showed a downregulation of the N-myc-interactor gene and presence of EMT. Furthermore, exposure of human primary bronchial epithelial cells to PsA resulted in downregulation of NMI and induction of an EMT phenotype while NMI upregulation resulted in attenuation of this PsA-induced EMT response. CONCLUSIONS: CLAD is associated with increased bacterial biomass and a Proteobacteria enriched airway microbiome and EMT. Proteobacteria such as PsA induces EMT in human bronchial epithelial cells via NMI, demonstrating a newly uncovered mechanism by which the microbiome induces cellular metaplasia.


Asunto(s)
Transición Epitelial-Mesenquimal/genética , Regulación de la Expresión Génica , Péptidos y Proteínas de Señalización Intracelular/genética , Trasplante de Pulmón/efectos adversos , Microbiota , Disfunción Primaria del Injerto/genética , ARN Ribosómico 16S/genética , Aloinjertos , Enfermedad Crónica , Regulación hacia Abajo , Células Epiteliales/metabolismo , Células Epiteliales/microbiología , Células Epiteliales/patología , Femenino , Estudios de Seguimiento , Humanos , Péptidos y Proteínas de Señalización Intracelular/biosíntesis , Masculino , Persona de Mediana Edad , Disfunción Primaria del Injerto/microbiología , Disfunción Primaria del Injerto/patología , Estudios Retrospectivos
9.
Database (Oxford) ; 20202020 01 01.
Artículo en Inglés | MEDLINE | ID: mdl-32090261

RESUMEN

Nipah virus (NiV) is an emerging and priority pathogen from the Paramyxoviridae family with a high fatality rate. It causes various diseases such as respiratory ailments and encephalitis and poses a great threat to humans and livestock. Despite various efforts, there is no approved antiviral treatment available. Therefore, to expedite and assist the research, we have developed an integrative resource NipahVR (http://bioinfo.imtech.res.in/manojk/nipahvr/) for the multi-targeted putative therapeutics and epitopes for NiV. It is structured into different sections, i.e. genomes, codon usage, phylogenomics, molecular diagnostic primers, therapeutics (siRNAs, sgRNAs, miRNAs) and vaccine epitopes (B-cell, CTL, MHC-I and -II binders). Most decisively, potentially efficient therapeutic regimens targeting different NiV proteins and genes were anticipated and projected. We hope this computational resource would be helpful in developing combating strategies against this deadly pathogen. Database URL: http://bioinfo.imtech.res.in/manojk/nipahvr/.


Asunto(s)
Bases de Datos Genéticas , Infecciones por Henipavirus , Virus Nipah , Animales , Antivirales , Epítopos/genética , Genoma Viral/genética , Infecciones por Henipavirus/tratamiento farmacológico , Infecciones por Henipavirus/virología , Humanos , Patología Molecular , Filogenia , ARN no Traducido/genética , ARN Viral/genética
10.
Front Pharmacol ; 10: 71, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-30809147

RESUMEN

Nipah virus (NiV) caused several outbreaks in Asian countries including the latest one from Kerala state of India. There is no drug available against NiV till now, despite its urgent requirement. In the current study, we have provided a computational one-stop solution for NiV inhibitors. We have developed the first "anti-Nipah" web resource, which comprising of a data repository, prediction method, and data visualization module. The database contains of 313 (181 unique) chemicals extracted from research articles and patents, which were tested for different strains of NiV isolated from various outbreaks. Moreover, the quantitative structure-activity relationship (QSAR) based regression predictors were developed using chemicals having half maximal inhibitory concentration (IC50). Predictive models were accomplished using support vector machine employing 10-fold cross validation technique. The overall predictor showed the Pearson's correlation coefficient of 0.82 on training/testing dataset. Likewise, it also performed equally well on the independent validation dataset. The robustness of the predictive model was confirmed by applicability domain (William's plot) and scatter plot between actual and predicted efficiencies. Further, the data visualization module from chemical clustering analysis displayed the diversity in the NiV inhibitors. Therefore, this web platform would be of immense help to the researchers working in developing effective inhibitors against NiV. The user-friendly web server is freely available on URL: http://bioinfo.imtech.res.in/manojk/antinipah/.

11.
Indian J Med Microbiol ; 37(1): 24-28, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31424006

RESUMEN

Introduction: Varicella outbreaks are known to occur in developing nations as vaccine coverage is still low. Material and Methods: In the present study, an institutional outbreak from Chandigarh, India, is reported wherein the utility of non-invasive samples such as saliva and urine was studied for the molecular diagnosis of varicella by conventional polymerase chain reaction (PCR), real-time PCR and real-time loop-mediated isothermal amplification (real-time LAMP). Results: The results of the present study showed that saliva and urine samples can be used for outbreak investigation of varicella compared to varicella-zoster virus DNA in vesicular swab samples with reasonable sensitivity. Conclusion: Thus, molecular techniques may be useful in the early identification of the outbreak and timely isolation, and the treatment of cases can further prevent its spread.


Asunto(s)
Varicela/diagnóstico , Varicela/epidemiología , Técnicas de Diagnóstico Molecular/métodos , Saliva/virología , Orina/virología , Adolescente , Anticuerpos Antivirales/sangre , Varicela/inmunología , Niño , Protección a la Infancia , ADN Viral/análisis , Brotes de Enfermedades/estadística & datos numéricos , Femenino , Instituciones de Salud , Herpesvirus Humano 3/genética , Herpesvirus Humano 3/inmunología , Herpesvirus Humano 3/aislamiento & purificación , Humanos , India/epidemiología , Masculino , Técnicas de Amplificación de Ácido Nucleico/métodos , Reacción en Cadena de la Polimerasa
13.
Viral Immunol ; 31(8): 589-593, 2018 10.
Artículo en Inglés | MEDLINE | ID: mdl-29792782

RESUMEN

The antiviral and immunomodulatory role of vitamin D has been shown in various viral infections. However, there is scanty literature available about the effect of vitamin D supplementation in herpes simplex virus-1 (HSV-1) infection. Therefore, the present study aimed to evaluate the role of two different forms of vitamin D: 25-hydroxyvitamin D3 (25D3) and 1,25-dihydroxyvitamin D3 (1,25D3) against HSV-1 in HeLa cells. The HeLa cells were supplemented with either 25D3 or 1,25D3 before HSV-1 infection and were studied after 6, 12, and 24 h postinfection (p.i.). The mRNA levels of toll-like receptors (TLRs), (2, 3, 4, 7, and 9), vitamin D signaling genes, and HSV-1 were studied using real-time PCR. The HSV-1 DNA load was estimated in culture supernatant. The supplementation of 25D3 and 1,25D3 significantly downregulated the mRNA levels of TLR2 (p < 0.0001) at 12 h p.i. The mRNA levels of TLR9 were found to be significantly downregulated in 1,25D3-supplemented cells at 12 h p.i. Furthermore, the significant downregulation was observed in HSV-1 titer in both 25D3- and 1,25D3-supplemented cells at 24 h p.i.(p < 0.0001). However, the effect of 25D3 supplementation persisted till 24 h p.i. with significant downregulation of TLR2 (p < 0.05) mRNA levels. The supplementation of both 25D3 and 1,25D3 before HSV-1 infection was found to downregulate the viral titer and TLR2 mRNA during the intial phase of infection. However, the effect of 25D3 supplementation was found to last for a longer duration compared with 1,25D3.


Asunto(s)
Antivirales/farmacología , Calcifediol/farmacología , Calcitriol/farmacología , Herpesvirus Humano 1/efectos de los fármacos , Factores Inmunológicos/farmacología , Células HeLa , Herpes Simple/tratamiento farmacológico , Herpes Simple/virología , Humanos , ARN Viral/efectos de los fármacos , Receptor Toll-Like 9/efectos de los fármacos , Receptor Toll-Like 9/genética , Receptores Toll-Like/efectos de los fármacos , Receptores Toll-Like/genética
14.
Eur J Drug Metab Pharmacokinet ; 43(5): 607-613, 2018 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-29577177

RESUMEN

BACKGROUND AND OBJECTIVES: The protective role of vitamin D supplementation has recently been shown to be present in various ocular inflammatory diseases. The oral supplementation of vitamin D may take time to achieve adequate levels in intraocular fluids. Therefore, the present study was performed to understand the ocular pharmacokinetics of 25-hydroxyvitamin D3 (25D3) in aqueous humor after weekly supplementation of 25D3 in rabbits. METHODS: A total of 21 rabbits were fed orally with 25D3 (7.22 µg/kg/week) for 8 weeks and 9th dose was given at the end of 8 weeks. The blood and aqueous humor samples were collected from ear vein and though anterior chamber paracentesis, respectively. The serum and aqueous humor samples were spiked with deuterium labeled internal standard and were extracted using liquid extraction method. Furthermore, the samples were derivatized and 25D3 estimation was performed using ultra performance liquid chromatography-tandem mass spectrometer (UHPLC-MS/MS). RESULTS: The 25D3 supplementation significantly increased the 25D3 levels in serum (78.5 ± 21.6 ng/ml) (mean ± SD) (p < 0.0001) and in aqueous humor (991.3 ± 180.6 pg/ml) (mean ± SD) (p < 0.0001) compared to baseline levels. The maximum concentration was achieved in serum after the 10th hour of supplementation of 1st and 9th dose, while the same was observed at the 24th hour in aqueous humor. CONCLUSION: The oral supplementation of 25D3 was found to significantly increase 25D3 levels in aqueous humor; however, the time required to achieve 25D3 concentration in aqueous humor was higher as compared to that in serum. Therefore, weekly oral supplementation of 25D3 may have a beneficial role in ocular diseases.


Asunto(s)
Humor Acuoso/metabolismo , Calcifediol/administración & dosificación , Calcifediol/farmacocinética , Cromatografía Líquida de Alta Presión/métodos , Suplementos Dietéticos , Espectrometría de Masas en Tándem , Administración Oral , Animales , Calcifediol/sangre , Esquema de Medicación , Conejos
15.
Indian J Med Microbiol ; 36(1): 113-115, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29735838

RESUMEN

Human adenovirus (HAdV) is a major cause of viral conjunctivitis. The various serotypes implicated in the causation are 3, 4, 8, 19 and 37. The present study aimed to know the circulating types of HAdV causing acute conjunctivitis in North India. A total of 23 conjunctival swabs were collected from patients with clinically suspected acute viral conjunctivitis during 2014-2015. The HAdV was implicated in the etiology in 65.2% of cases. The sequencing of representative samples using hexon gene suggests the presence of serotype 8 and 4. The serotype eight sequences showed 99%-100% similarity with other Indian strains. The phylogenetic analysis showed that the current circulating serotypes, responsible for conjunctivitis, belonged to epidemic keratoconjunctivitis strains.


Asunto(s)
Infecciones por Adenovirus Humanos/epidemiología , Adenovirus Humanos/clasificación , Adenovirus Humanos/aislamiento & purificación , Conjuntivitis Viral/epidemiología , Adenovirus Humanos/genética , Adulto , Secuencia de Bases , Conjuntivitis Viral/virología , ADN Viral/genética , Femenino , Genoma Viral , Humanos , India/epidemiología , Masculino , Epidemiología Molecular , Filogenia , Análisis de Secuencia de ADN
16.
Int J Appl Basic Med Res ; 7(1): 3-9, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28251100

RESUMEN

Recently, a number of techniques have been approved for quantification of viral nucleic acids in clinical samples. Viral load (VL) tests have considerable importance in the management of patients and are widely used in routine diagnosis. In clinical virology, VL testing are important to monitor the antiviral treatment, to initiate preemptive therapy, to understand pathogenesis, and to evaluate the infectivity. These tests have now become a part of many diagnostic and treatment guidelines. Considering the various challenges for in-house viral testing related to the standardization, validation, and precision; they are gradually being replaced by the United States Food and Drug Administration (US FDA) cleared tests. This review summarizes the various viral quantification methods and also discusses the clinical applicability of these in human immunodeficiency virus, Hepatitis B virus, Hepatitis C virus, Cytomegalovirus, and Epstein Barr virus infected patients. Further the challenges and future perspectives of VL testing have also been discussed.

17.
Indian J Med Microbiol ; 35(1): 113-115, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28303830

RESUMEN

Detecting high-risk-human papillomavirus (HPV) types has become an integral part of the cervical cancer screening programmes. This study aimed to develop a multiplex polymerase chain reaction (PCR) for identification of HPV types 16 and 18 along with the beta globin gene in formalin-fixed and paraffin-embedded cervical biopsy specimens. A total of 59 samples from patients with cervical abnormalities were tested. HPV 16 positivity was 50% in cervical cancers and 52.9% in cervical intraepithelial neoplasia. Our multiplex PCR protocol can be used as a simple and cost-effective tool for high-risk-HPV detection in cervical cancer screening programmes.


Asunto(s)
Cuello del Útero/virología , Papillomavirus Humano 16/aislamiento & purificación , Papillomavirus Humano 18/aislamiento & purificación , Reacción en Cadena de la Polimerasa Multiplex/métodos , Infecciones por Papillomavirus/diagnóstico , Patología Molecular/métodos , Carcinoma in Situ/diagnóstico , Carcinoma in Situ/virología , Costos y Análisis de Costo , Detección Precoz del Cáncer/métodos , Femenino , Papillomavirus Humano 16/genética , Papillomavirus Humano 18/genética , Humanos , Persona de Mediana Edad , Infecciones por Papillomavirus/virología , Fijación del Tejido , Neoplasias del Cuello Uterino/diagnóstico , Neoplasias del Cuello Uterino/virología
18.
Diagn Microbiol Infect Dis ; 89(3): 191-196, 2017 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-28911798

RESUMEN

Infectious uveitis is a vision threatening inflammatory ocular disease wherein early diagnosis may prevent the loss of vision. The purpose of this study was to develop a multiplex real-time PCR for the diagnosis of Herpes simplex virus-1, Varicella zoster virus, cytomegalovirus and Toxoplasma gondii in patients with suspected infectious uveitis. A total of 126 intraocular samples (aqueous and vitreous humor) were collected and subjected to multiplex real-time PCR. Overall 26.2% (33/126) patients were found to be positive for one or more of the pathogens tested. The overall positivity for VZV, HSV, CMV and T. gondii was found to be 16 (12.7%), 7 (5.6%), 5 (3.9%), and 9 (7.1%); with mean pathogen load of 5.07×105, 9.5×104, 1.08×104 and 394 (copies/µl) respectively. The development of highly sensitive and specific assay for early differentiation of pathogens is important for the early initiation of treatment thereby preventing irreversible damage to the ocular structures.


Asunto(s)
Citomegalovirus/aislamiento & purificación , Herpesvirus Humano 1/aislamiento & purificación , Herpesvirus Humano 3/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Toxoplasma/aislamiento & purificación , Uveítis/diagnóstico , Citomegalovirus/genética , ADN Protozoario/genética , ADN Protozoario/aislamiento & purificación , ADN Viral/genética , ADN Viral/aislamiento & purificación , Herpesvirus Humano 1/genética , Herpesvirus Humano 3/genética , Humanos , Sensibilidad y Especificidad , Toxoplasma/genética , Toxoplasmosis/diagnóstico , Toxoplasmosis/parasitología , Uveítis/parasitología , Uveítis/virología , Virosis/diagnóstico , Virosis/virología
19.
Indian J Med Microbiol ; 35(3): 417-420, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-29063890

RESUMEN

Outbreaks of varicella are reported when susceptible population accumulates. This study reports a chickenpox outbreak in Burail in August 2014, wherein 20 laboratory-confirmed cases were identified by the detection of varicella zoster virus (VZV) DNA and VZV IgM antibodies. The viral load between vesicular swabs and serum samples from 8 patients with active lesions was found to have good correlation and further also related with disease severity. Real-time polymerase chain reaction can be useful for early diagnosis of an outbreak and vesicular swab can be used as a less invasive sample for assessing the disease severity.


Asunto(s)
Varicela/epidemiología , Pruebas Diagnósticas de Rutina/métodos , Brotes de Enfermedades , Epidemiología Molecular/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Adolescente , Adulto , Anticuerpos Antivirales/sangre , Niño , Preescolar , ADN Viral/sangre , ADN Viral/genética , Femenino , Humanos , Inmunoglobulina G/sangre , India/epidemiología , Lactante , Masculino , Adulto Joven
20.
Diagn Microbiol Infect Dis ; 85(4): 477-81, 2016 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-27236713

RESUMEN

Congenital cataract has the potential for inhibiting early visual development. Intrauterine infections with Rubella virus, Herpes simplex virus (HSV) and Toxoplasma gondii plays an important role in the development of congenital cataract. The study included 120 children under the age of 6 years presenting with congenital cataract and diagnosed using serology and polymerase chain reaction (PCR). The IgM positivity for rubella, HSV, T. gondii was found to be 5.8%, 1.6% and 8.3% respectively. The overall PCR positivity was found to be 40(33.3%), 25 (20.8%) and 39 (32.5%) for rubella, HSV and T. gondii with mean copy number of 1599 copies/µL; 1716 copies/µL and 1503 copies/µL respectively. Infective etiology significantly contributes to the causation of congenital cataract particularly for rubella virus which is a potentially eradicable disease. This study provides an epidemiological data for rubella, HSV and T. gondii in children with congenital cataract and highlights the need to introduce rubella vaccine in the National Immunization Programme of India.


Asunto(s)
Catarata/congénito , Virus de la Rubéola/aislamiento & purificación , Simplexvirus/aislamiento & purificación , Toxoplasma/aislamiento & purificación , Anticuerpos Antiprotozoarios/sangre , Anticuerpos Antivirales/sangre , Niño , Preescolar , ADN Protozoario/análisis , ADN Protozoario/genética , ADN Viral/análisis , ADN Viral/genética , Femenino , Humanos , Inmunoensayo , Inmunoglobulina M/sangre , India , Cristalino/parasitología , Cristalino/virología , Masculino , Reacción en Cadena de la Polimerasa , Estudios Prospectivos , ARN Viral/análisis , ARN Viral/genética , Centros de Atención Terciaria
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