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Background: Hospital milieu monitoring is an essential component for controlling healthcare associated infections (HCAIs) as it serves as the reservoir for pathogenic microbes. Aim of this study was to identify the bacterial load in Intensive care units (ICU) and Operation theaters (OT) air and water sources of selected tertiary care hospitals. Material & Methods: The study was organized in Microbiology department, BIRDEM General Hospital. A sum total 28 air samples & 6 water samples were collected from three selected hospitals and those were processed according to the set of protocols. Results: From air sampling, highest load of bacteria was found 480 CFU/dm²/hr in Hospital C ICU, 38.40 ± 9.99 CFU/dm²/hr in pre-OT samples & 218.2±43.35 CFU/dm²/hr in intra OT samples of Hospital C. From water sampling, unacceptable level of coliforms was found in all three hospitals. Among the non-pathogens, 24% – 37% Micrococcus spp. (normal flora) and 2% -18% Bacillus spp. (contaminants) were found in the OTs. Whereas pathogens found were Acinetobacter spp. (20.7%) followed by Pseudomonas spp. (19.4%), Klebsiella spp. (12.1%) & S. aureus (9.2%) in the ICUs. Conclusion: It could be deduced from the study that environmental sources such as air and water contaminations with multidrug resistant pathogens are an ultimate risk factor for all related to the healthcare settings, specially the indoor patients.
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Background: The anterior cruciate ligament (ACL) is the most frequently disrupted ligament of the knee. Arthroscopic ACL reconstruction is one of the most common orthopaedic procedures now-a-days and has become the gold standard method of treatment in complete ACL tear. Though the choice of graft for ACL reconstruction and the methods of its fixation are matters of debate, hamstring tendon is considered as the favorable graft. Common options for the choice of graft are quadrupled semitendinosus-gracilis and tripled semitendinosus auto graft. Aim of our study is to compare the outcome of arthroscopic ACL reconstruction by quadrupled semitendinosus-gracilis and tripled semitendinosus auto graft.Material & Methods:In this prospective interventional study purposive sampling was done. Sixty patients with complete ACL tear were included in this study from May 2017 to August 2019 at NITOR, strictly considering the inclusion and exclusion criteria. Thirty of them treated with quadrupled semitendinosus-gracilis (group 1) and thirty with tripled semitendinosus (group 2) auto graft. Evaluation by Lysholm knee score was done before and after surgery. Final outcome was evaluated at 24th week post-operatively.Results:Preoperative Lysholm Knee Score was almost similar in group 1 (63.2�1) and in group 2 (63.1�5). This difference was not statistically significant (p=0.582). Postoperative Lysholm Knee Score at 24th week was slightly higher in group 1 (95.0�0) in comparison to group 2 (94.4�2). This difference was also not statistically significant (p=0.361). But excellent functional outcome was more in group 1 (80.0%) in comparison to group 2 (73.3%) whereas no poor or fair outcome was reported in group 1 but fair outcome was reported in 6.7% of patients in group 2. Overall complication rate was 6.67% with no significant intergroup difference.Conclusions:Quadrupled semitendinosus-gracilis auto graft has yielded more excellent results than tripled semitendinosus graft. But as this difference was not significant, it can be said that, both the procedure can be performed for arthroscopic reconstruction of ACL.
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Introduction: For the rapid and accurate genetic identification and authentication of living organisms, improved random amplified polymorphic DNA (RAPD) fragment based development of sequence-characterized amplified region (SCAR) markers is an important genetic technique. Objective: This study aimed to develop SCAR markers for perennial herb Eclipta prostrate (E. prostrate). Methods: Here the RAPD fragments by improved RAPD amplification with primers A11 and N-7 for E. prostrate were cloned into pGEX-T vector, and PCR amplification identified the positive clones. After the enzymatic digestion, they were sequenced with Sanger sequencing. Results: Two SCAR markers were developed, which were very specific to E. prostrate, not found in Penthorum chinense Pursh(P. chinense). The nucleotide sequence search by BLAST GenBank database showed that they are novel in E. prostrate, therefore they were deposited in Genbank with accession number KX671034, KX671035. The markers did not show any identity to other species. Conclusions: Thus, in this study two specific SCAR markers were developed for genetically distinguishing and identifying the plant species E. prostrate from herb P. chinense and others.
Introducción: Verificación genética del arbusto Eclipta prostrate (Asteraceae) (Para la identificación y verificación genética rápida y precisa de organismos vivos, el uso de fragmentos de ADN polimórfico amplificado aleatoriamente (RAPD) mejorado de marcadores de región amplificada caracterizada por secuencia (SCAR) es una técnica genética importante. Objetivo: Este estudio tuvo como objetivo desarrollar marcadores SCAR para la hierba perenne Eclipta postrate (E. postrate). Métodos: En este estudio os fragmentos RAPD mediante amplificación RAPD mejorada con los cebadores A11 y N-7 para E. postrate se clonaron en el vector pGEX-T, y la amplificación por PCR identificó los clones positivos. Después de la digestión enzimática, se realizó una secuenciación Sanger. Resultados: Se desarrollaron dos marcadores SCAR, muy específicos para E. postrate, que no se encuentran en Penthorum chinense Pursh (P. chinense). La búsqueda de las secuencias de nucleótidos con BLAST en GenBank mostró que son nuevos en E. postrate, por lo que fueron depositados en Genbank con los números de acceso: KX671034 y KX671035. Los marcadores no mostraron ninguna identidad a otras especies. Conclusiones: En este estudio se desarrollaron dos marcadores SCAR específicos para distinguir e identificar genéticamente la especie de planta E. postrate de la hierba P. chinense y otras.
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Objective: To analyze the phytochemical constituents responsible for the plausible antioxidant effect of methanolic extract of the seed, pulp and peel of Baccaurea ramiflora Lour. Methods: Fresh seed, pulp, and peel of Baccaurea ramiflora fruits were extracted with methanol (MEBRse, MEBRpu, MEBRpe) and evaluated by phytochemical analysis for their content of innumerable metabolites (primary and secondary) viz. carbohydrates, alkaloids, glycosides, tannins, phenols, terpenoids, flavonoids, proteins, and fixed oils. The antioxidant efficacy was assessed through different assay methods viz. 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, total antioxidant capacity (TAC) and reducing power capacity (RPC). Estimation of total phenolic content (TPC), and total flavonoid content (TFC) was also done to confirm the presence of these phytochemicals. Results: It was revealed from the phytochemical analysis of MEBRse that alkaloids, glycosides, carbohydrates, phenols, and flavonoids were present, while that of MEBRpu showed the existence of carbohydrates, proteins, alkaloids, glycosides, phenols, saponins, flavonoids, and fixed oils. Presence of carbohydrates, alkaloids, phenols, tannins, flavonoids, and terpenoids were found in the MEBRpe. A significant antioxidant activity was revealed by the MEBRpu [EC
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Objective:To analyze the phytochemical constituents responsible for the plausible antioxidant effect of methanolic extract of the seed, pulp and peel of Baccaurea ramiflora Lour.Methods:Fresh seed, pulp, and peel of Baccaurea ramiflora fruits were extracted with methanol (MEBRse, MEBRpu, MEBRpe) and evaluated by phytochemical analysis for their content of innumerable metabolites (primary and secondary) viz. carbohydrates, alkaloids, glycosides, tannins, phenols, terpenoids, flavonoids, proteins, and fixed oils. The antioxidant efficacy was assessed through different assay methods viz. 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, total antioxidant capacity (TAC) and reducing power capacity (RPC). Estimation of total phenolic content (TPC), and total flavonoid content (TFC) was also done to confirm the presence of these phytochemicals.Results:It was revealed from the phytochemical analysis of MEBRse that alkaloids, glycosides, carbohydrates, phenols, and flavonoids were present, while that of MEBRpu showed the existence of carbohydrates, proteins, alkaloids, glycosides, phenols, saponins, flavonoids, and fixed oils. Presence of carbohydrates, alkaloids, phenols, tannins, flavonoids, and terpenoids were found in the MEBRpe. A significant antioxidant activity was revealed by the MEBRpu [ECConclusions:This study suggests that MEBRpu has a significantly higher antioxidant property than MEBRpe and MEBRse. These extracts might be advantageous in prevention or decelerating the progress of different diseases related to oxidative-stress/damage. Moreover, detailed analysis of these extracts is required to identify the presence of promising compound(s) responsible for their antioxidant activity.
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Background: Penthorum chinense Pursh (P. chinense) is a well-known traditional Chinese medicine (TCM) plant, which has long been used for the prevention and treatment of hepatic diseases. This study aimed to genetically characterize the varieties of P. chinense from different geographic localities of China by random amplification of polymorphic DNA (RAPD)-PCR technique and verified with inter-simple sequence repeat (ISSR) markers. Results: The P. chinense samples were collected from nine different geographic localities. Previously improved RAPD and ISSR markers were utilized for genetic analysis using DNA amplification. The genetic relationship dendrogram was obtained by conducting cluster analysis to the similarity coefficient of improved RAPD and ISSR markers. Improved RAPD yielded 185 scorable amplified products, of which 68.6% of the bands were polymorphic, with an average amplification of 9.25 bands per primer. The ISSR markers revealed 156 alleles with 7.8 bands per primers, where 59.7% bands were polymorphic. Furthermore, the similarity coefficient ranges of RAPD and ISSR markers were 0.710.91 and 0.660.89, respectively. Conclusions: This study indicated that improved RAPD and ISSR methods are useful tools for evaluating the genetic diversity and characterizing P. chinense. Our findings can provide the theoretical basis for cultivar identification, standardization, and molecular-assisted breeding of P. chinense for medicinal use.
Subject(s)
Plants, Medicinal/genetics , Magnoliopsida/genetics , Polymorphism, Genetic , Genetic Variation , Genetic Markers , China , DNA, Plant/genetics , Random Amplified Polymorphic DNA Technique , Microsatellite Repeats , Medicine, Chinese TraditionalABSTRACT
Gardenia jasminoides is a common garden medicinal plant known for its anticancer, anti-inflammatory, anti-thrombic, anti-fibrotic, antiviral, hepatoprotective, lung-protective, renal-protective, retina-protective and neuroprotective activities. It is found in several regions of the world, including China, but information about its genetic characteristics is limited. Here, we employed an improved method of random amplified polymorphic DNA (RAPD) analysis (with increased RAMP time) to investigate the genetic link between G. jasminoides samples collected from six different regions of Southern China. Total 26 RAPD primers were selected randomly, among which 23 primers generated reproducible polymorphic amplification bands. A total of 174 bands were obtained, where each primer had amplified 5-13 bands with an average of 7.56 bands per primer. The band size ranged approximately 150-2200 bp. Cluster dendrogram was obtained based on the improved RAPD amplification profiles, which showed that the similarity coefficients among six varieties of G. jasminoides ranged 0.67-0.88. To our knowledge, this is the first report of genetic characterization of G. jasminoides using improved RAPD analysis, which may be useful for the preservation of genetic diversity and identification of Gardenia population.
Subject(s)
China , /genetics , /isolation & purification , Electrophoresis, Agar Gel , Gardenia/classification , Gardenia/genetics , Gene Flow , Genetic Variation , Plants, Medicinal/classification , Plants, Medicinal/genetics , Random Amplified Polymorphic DNA Technique/methods , Reproductive IsolationABSTRACT
Genetic diversity within a species is a common feature, which plays a vital role in its survival and adaptability, and is important for the identification and authentication of a species. Lonicera japonica is a traditionally used medicinal plant, which have been recently genetically characterized by an improved ran- dom amplified polymorphic DNA (RAPD) analysis. In this study, the molecular markers on the basis of these RAPD fragments have been developed to identify specific L. japonica variety. The DNAs were extracted from fresh young leaves of different samples of L. japonica collected from Shenzhen, Yichang, Leshan, Emei and Loudi, China. The DNA materials were amplified using improved RAPD PCR. Different RAPD bands were excised, cloned and developed for stable sequence-characterized amplified region (SCAR) markers with differ- ent species. Two SCAR markers, JYH3-3 and JYH4-3, have been successfully cloned from improved RAPD fragments. The SCAR marker JYH3-3 was found specific for all of the L. japonica samples collected from the different regions, and another marker JYH 4-3 was strictly specific to the Shenzhen sample from Guangdong province, which is geographically distant from Hubei, Sichuan and Hunan Provinces (source of other L. japonica samples). The marker JYH3-3 was found as specific molecular marker for the identification of L. japonica, while JYH4-3 was found as molecular marker strictly specific for the Shenzhen sample. The developed SCAR mark- ers might serve as more specific molecular markers for L. japonica variety authentication. The combination of improved RAPD analysis and SCAR marker development have resulted useful tools to study the genetic variety of any organism, which we have successfully applied here in L. japonica.
La diversidad genética dentro de una especie es una característica común, que juega un papel vital en su supervivencia y adaptabilidad, y es importante para la identificación y la autenticación de una especie. Lonicera japonica es una planta medicinal utilizada tradicionalmente, que han sido recientemente caracterizada genéticamente por amplificación aleatoria mejorada de ADN polimórfico (RAPD). En este estudio, los marcadores moleculares basados en estos fragmentos de RAPD se han desarrollado para identificar una variedad específica de L. japonica. Los ADN se extrajeron de las hojas jóvenes frescas de diferentes muestras de L. japonica recogidas de Shenzhen, Yichang, Leshan, Emei y Loudi, China. Los materiales de ADN fueron amplificados utilizando el RAPD PCR mejorado. Diferentes bandas RAPD fueron extraídas, clonadas y desarrolladas para las regiones amplificadas de secuencia conocida (SCAR) con marcado- res de diferentes especies. Dos marcadores SCAR, JYH3-3 y JYH4-3, se clonaron con éxito de los RAPD mejorados. El marcador SCAR JYH3-3 se encontró específico para todas las muestras de L. japonica recolectadas en las diferentes regiones, mientras que el otro marcador JYH4-3 era estrictamente específico para la muestra de Shenzhen de la provincia de Guangdong, que está geográficamente distante de Hubei, Sichuan y Provincias Hunan (fuente de otras muestras de L. japonica). Se encontró que JYH3-3 es un marcador molecular específico para la identificación de L. japonica, mientras que JYH4-3 se encontró como marcador molecular estrictamente específico para la muestra de Shenzhen. Los marcadores SCAR desarrollados podrían servir como marcadores moleculares más específicos para la autenticación de la variedad L. japonica. La combi- nación de RAPD mejorado y el desarrollo del marcador SCAR han dado como resultado herramientas útiles para el estudio de la variedad genética de cualquier organismo, que hemos aplicado con éxito en L. japonica.
Subject(s)
Cloning, Molecular/methods , Lonicera/genetics , China , Genetic Markers , Lonicera/classification , Polymerase Chain Reaction , Random Amplified Polymorphic DNA TechniqueABSTRACT
BACKGROUND: We investigated whether wogonin and apigenin significantly affect the epidermal growth factor receptor (EGFR) signaling pathway involved in MUC5AC mucin gene expression, and production from cultured airway epithelial cells; this was based on our previous report that apigenin and wogonin suppressed MUC5AC mucin gene expression and production from human airway epithelial cells. METHODS: Confluent NCI-H292 cells were pretreated with wogonin or apigenin for 15 minutes or 24 hours and then stimulated with epidermal growth factor (EGF) for 24 hours or the indicated periods. RESULTS: We found that incubation of NCI-H292 cells with wogonin or apigenin inhibited the phosphorylation of EGFR. The downstream signals of EGFR such as phosphorylation of MEK1/2 and ERK1/2 were also inhibited by wogonin or apigenin. CONCLUSION: The results suggest that wogonin and apigenin inhibits EGFR signaling pathway, which may explain how they inhibit MUC5AC mucin gene expression and production induced by EGF.
Subject(s)
Humans , Apigenin , Epidermal Growth Factor , Epithelial Cells , Gene Expression , Mucins , Phosphorylation , ErbB ReceptorsABSTRACT
BACKGROUND: We investigated whether prunetin significantly affects tumor necrosis factor-alpha (TNF-alpha)-induced MUC5AC mucin gene expression, production, inhibitory kappa B (IkappaB) degradation and nuclear factor kappa B (NF-kappaB) p65 translocation in human airway epithelial cells. METHODS: Confluent NCI-H292 cells were pretreated with prunetin for 30 minutes and then stimulated with TNF-alpha for 24 hours or the indicated periods. MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The effect of prunetin on TNF-alpha-induced degradation of IkappaB and translocation of NF-kappaB p65 was investigated by western blot analysis. RESULTS: We found that incubation of NCI-H292 cells with prunetin significantly inhibited mucin production and down-regulated the MUC5AC gene expression induced by TNF-alpha. Prunetin inhibited TNF-alpha-induced degradation of IkappaB and translocation of NF-kappaB p65. CONCLUSION: This result suggests that prunetin inhibits the NF-kappaB signaling pathway, which may explain its role in the inhibition of MUC5AC mucin gene expression and production regulated by the NF-kappaB signaling pathway.
Subject(s)
Humans , Blotting, Western , Enzyme-Linked Immunosorbent Assay , Epithelial Cells , Gene Expression , Isoflavones , Mucin 5AC , Mucins , NF-kappa B , Polymerase Chain Reaction , Reverse Transcription , Tumor Necrosis Factor-alphaABSTRACT
BACKGROUND: We investigated whether chrysin affected MUC5AC mucin production and gene expression induced by phorbol ester (phorbol 12-myristate 13-acetate, PMA) or epidermal growth factor (EGF) from human airway epithelial cells. METHODS: Confluent NCI-H292 cells were pretreated with varying concentrations of chrysin for 30 minutes, and were then stimulated with PMA and EGF for 24 hours, respectively. MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay. RESULTS: Concentrations of 10microM and 100microM chrysin were found to inhibit the production of MUC5AC mucin protein induced by PMA; A concentration of 100microM chrysin also inhibited the production of MUC5AC mucin protein induced by EGF; 100microM chrysin inhibited the expression of MUC5AC mucin gene induced by PMA or EGF. The cytotoxicity of chrysin was checked by lactate dehydrogenase assay, and there was no cytotoxic effect observed for chrysin. CONCLUSION: These results suggest that chrysin can inhibit mucin gene expression and the production of mucin protein by directly acting on airway epithelial cells.