Phytochemical profiling and evaluation of modified resazurin microtiter plate assay of the roots of Trillium govanianum.

Trillium govanianum Wall. ex D. Don (Melanthiaceae alt. Trilliaceae), is native to the Himalayas. The present study, for the first time, was undertaken to explore the antimicrobial potential, to determine the minimum inhibitory concentration (MIC) values of the methanol extract of the roots of Trillium govanianum and its solid phase extraction (SPE) fractions by using resazurin microtiter assay (REMA) against Gram positive and Gram negative bacterial registered strains and to carry out phytochemical analysis. The remarkable amount of gallic acid equivalent phenolic and quercetin equivalent flavonoid content was manifested by MeOH extract (20.27 ± 3.03 mg GAE/g DW and 9.25 ± 0.50 mg QE/g DW respectively). The GC/MS analysis revealed the presence saturated and unsaturated components. Considerable level of antibacterial potential against Gram-positive bacteria (MIC: 2.5-0.009 mg/mL) than against Gram-negative bacteria (MIC: 2.5-0.165 mg/mL) were observed. The use of microtiter plates has the advantage of lower cost, fast and quantitative results.

Gold nanoworms: Optical properties and simultaneous SERS and fluorescence enhancement.

Fabrication of platforms for efficient Raman and fluorescence enhancement is very attractive for plasmon-based molecular sensing. For superior sensitivity, the plasmonic constituents of such platforms should be effective electromagnetic field enhancers. Furthermore, nanoparticles having plasmon peak in the spectral range of therapeutic window are superior for biomedical application. Herein, we show that worm-shaped Au nanoparticles can be used for simultaneous surface enhanced Raman scattering (SERS) and metal enhanced fluorescence (MEF). Gold nanoworms (Au-NWs) with an intense plasmon absorption in the therapeutic window were synthesized using a facile single-stepped recipe. Electron microscopy imaging revealed that Au-NWs have a non-uniform surface. Owing to their special morphology, three distinct plasmon bands were seen in the experimental spectrum. The presence of three plasmon peaks was also verified by finite element based simulations. The simulation results further show that Au-NWs can provide intense near field enhancement for multiple excitation wavelengths. As a proof of concept, we have used Au-NWs based platforms for simultaneous enhancement of fluorescence and Raman signal of rhodamine 6G (R6G) dye molecule.

Liquid Chromatography Mass Spectrometry Analysis and Cytotoxicity of Asparagus adscendens Roots against Human Cancer Cell Lines.

BACKGROUND: Asparagus adscendens Roxb. (Asparagaceae), is native to the Himalayas. This plant has been used in the prevention and effective treatment of various forms of cancers. OBJECTIVE: This paper reports, for the first time, on the cytotoxicity of the methanol (MeOH) extract of the roots of A. adscendens and its solid-phase extraction (SPE) fractions against four human carcinoma cell lines and LC-ESI-QTOF-MS analysis of the SPE fractions. MATERIALS AND METHODS: Finely powdered roots of A. adscendens were macerated in methanol and extracted through SPE using gradient solvent system (water: methanol) proceeded for analysis on LC-ESI-QTOF-MS and cytotoxicity against four human carcinoma cell lines: breast (MCF7), liver (HEPG2), lung (A549), and urinary bladder (EJ138), using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay. RESULTS: The MeOH extract and four SPE fractions exhibited cytotoxicity against all cell lines with the IC50 values ranging from 6 to 79 µg/mL. As observed in other Asparagus species, the presence of saponins and sapogenins in the SPE fractions was evident in the liquid chromatography-mass spectrometry data. CONCLUSION: It is reasonable to assume that the cytotoxicity of the MeOH extract of the roots of A. adscendens and its SPE fractions, at least partly, due to the presence of saponins and their aglycones. This suggests that A. adscendens could be exploited as a potential source of cytotoxic compounds with putative anticancer potential. SUMMARY: The MeOH extract and all solid-phase extraction (SPE) fractions exhibited various levels of cytotoxicity against all cell lines with the IC50 values ranging from 6 to 79 µg/mLThe presence of saponins and sapogenins in the SPE fractions was evident in the Liquid chromatography-mass spectrometry dataDue to the presence of saponins and their aglycones, suggest that A. adscendens could be exploited as a potential source of cytotoxic compounds with putative anticancer potential. Abbreviation used: SPE: Solid-phase extraction, MCF7: Breast cancer cell line, HEPG2: Liver cancer cell line, A549: Lung liver cancer cell line, EJ138: Urinary bladder cancer cell line, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide, LC-MS: Liquid chromatography-mass spectrometry.

Evaluation of resazurin microtiter plate assay and HPLC- photodiode array analysis of the roots of Asparagus adscendens.

Asparagus adscendens Roxb. (Asparagaceae), is native to the Himalayas. The present study, for the first time, was undertaken to explore the antimicrobial potential, to determine the minimum inhibitory concentration (MIC) values of the methanol extract of the roots of A. adscendens and its solid-phase extraction (SPE) fractions using resazurin microtitre assay against Gram-positive and negative bacterial-registered strains and to carry out HPLC-photodiode array analysis of the SPE fractions. The methanol extract and all SPE exhibited considerable level of antibacterial potential against Gram-positive bacteria (MIC: 2.5-0.009 mg/mL) than against Gram-negative bacteria (MIC: 1.25-2.5 mg/mL). The use of microtitre plates has the advantage of lower cost, fast and quantitative results. Like other Asparagus species, the presence of phenolic compounds in all SPE fractions was evident in the HPLC-PDA data.

The effects of gait training with body weight support (BWS) with no body weight support (no-BWS) in stroke patients.

The purpose of this study was to measure the clinical outcomes for patients with stroke after gait training with body weight support (BWS) and with no body weight support (no-BWS).Experimental group was trained to walk by a BWS system with overhead harness (BWS group), and Control group was trained with full weight bearing walk on their lower extremities. Treatment session comprised of six weeks training. Treatment outcomes were assessed on the basis of Timed 10 Meter Walk Test, Timed Get Up and Go Test and Dynamic Gait Index. There was a significant (P<0.05) difference in BWS and NBWS for Dynamic Gait Index, Timed Get Up and Go Test, Timed 10 Meter Walk Test (Self-Selected Velocity), and Timed 10 Meter Walk Test (Fast-Velocity). Training of gait in stroke patients while a percentage of their body weight supported by a harness, resulted in better walking abilities than the Training of gait while full weight was placed on patient's lower extremities.

Reliability of the universal goniometer for assessing active cervical range of motion in asymptomatic healthy persons.

OBJECTIVE: To determine within-rater and between-rater reliability of the universal goniometer (UG) for measuring active cervical range of motion (ACROM) in asymptomatic healthy subjects. METHODS: Nineteen healthy subjects were tested in an identical seated position. Two raters used UG to measure active cervical movements of flexion, extension, right side flexion, left side flexion, right rotation and left rotation. Each motion was measured twice by each of the two raters and was re-measured all over again after one week. Data analysis was performed using the intraclass correlation coefficient (ICC). RESULTS: The results demonstrated excellent within-session (ICC2,1 = 0.83 to 0.98) and between-session (ICC2,2 = 0.79 to 0.97) intra-rater reliability and excellent inter-rater reliability (ICC2,2 = 0.79 to 0.92). CONCLUSION: Considering above results it is concluded that UG is a reliable tool for assessing ACROM in a clinical setting for healthy subjects.

Socio-economic determinants of the awareness and adoption of citrus production practices in Pakistan / Determinantes socioeconômicos da conscientização e adoção de práticas de produção de citros no Paquistão

Citrus is the leading fruit of Pakistan and famous worldwide especially kinnow cultivar because of its pleasant taste and remarkable quality. The yield of citrus per hectare in Pakistan is almost half of potential due to non-adoption of recommended horticultural practices by citrus growers. Adopting a decision regarding the improvement of practices is usually influenced by various factors including farmers' socio-economic attributes. In order to determine the relationship between socio-economic aspects and the awareness and adoption of recommended citrus production practices the present study was carried out in Sargodha district from central Punjab, Pakistan. The Study was based upon cross sectional survey research design due to availability of sampling frame, probability (random sampling) was applied for sample selection. Through random sampling, 120 citrus growers were selected as sample. Structured questionnaire administered through interview was used as a research instrument. Analysis of the data collected from the targeted citrus growers revealed a highly significant influence of education on awareness and adoption. Moreover, significant association was found between citrus cultivation area and awareness and adoption of improved practices. Age also showed significant association with awareness and adoption. Moreover, dominancy of middle aged farmers and illiteracyin the study area strongly point the need of provision of formal and non-formal education and training program for farmers. Young generation needs to be focused and reorientation of youth clubs may help in better way to gain the utmost outcome.

Citrus é fruto líder do Paquistão e famoso no mundo inteiro, especialmente Kinnow, cultivado por causa do sabor agradável e de qualidade notável. Por hectare, o rendimento de citrus no Paquistão é quase metade do potencial, devido à não-adoção de práticas de horticultura recomendadas por citricultores. A adoção da decisão sobre práticas melhoradas geralmente é influenciada por vários fatores, incluindo atributos socioeconômicos dos agricultores. A fim de determinar a relação entre os aspectos socioeconômicos e a consciência e adoção de práticas de produção de citros recomendadas presentes, este estudo foi realizado no distrito de Sargodha do centro de Punjab, Paquistão. O estudo foi baseado em projeto de pesquisa do estudo transversal e, devido à disponibilidade de amostragem, amostragem quadro probabilidade (amostragem aleatória) foi aplicada para a seleção da amostra. Por meio de amostragem aleatória, 120 citricultores foram selecionados como amostra. Questionário estruturado, aplicado por meio de entrevista, foi utilizado como um instrumento de pesquisa. A análise dos dados coletados dos citricultores direcionados revelou uma influência altamente significativa da educação sobre a conscientização e adoção. Além disso, foi encontrada associação significativa entre a área de cultivo de citrinos e conscientização e adoção de práticas melhoradas. A idade dos produtores também mostrou associação significativa com a conscientização e adoção. Além disso, a dominância dos agricultores de meia idade e a falta de conhecimento na área de estudo apontam fortemente para a necessidade de oferta de educação formal e não-formal e de formação para os agricultores. Necessidades de geração de jovens a ser focado e reorientação dos clubes de jovens pode contribuir para melhor maneira de ganhar o máximo resultado.

Abundance and dynamics of antibiotic resistance genes and integrons in lake sediment microcosms.

Antibiotic resistance in bacteria causing disease is an ever growing threat to the world. Recently, environmental bacteria have become established as important both as sources of antibiotic resistance genes and in disseminating resistance genes. Low levels of antibiotics and other pharmaceuticals are regularly released into water environments via wastewater, and the concern is that such environmental contamination may serve to create hotspots for antibiotic resistance gene selection and dissemination. In this study, microcosms were created from water and sediments gathered from a lake in Sweden only lightly affected by human activities. The microcosms were exposed to a mixture of antibiotics of varying environmentally relevant concentrations (i.e., concentrations commonly encountered in wastewaters) in order to investigate the effect of low levels of antibiotics on antibiotic resistance gene abundances and dynamics in a previously uncontaminated environment. Antibiotic concentrations were measured using liquid chromatography-tandem mass spectrometry. Abundances of seven antibiotic resistance genes and the class 1 integron integrase gene, intI1, were quantified using real-time PCR. Resistance genes sulI and ermB were quantified in the microcosm sediments with mean abundances 5 and 15 gene copies/10(6) 16S rRNA gene copies, respectively. Class 1 integrons were determined in the sediments with a mean concentration of 3.8 × 10(4) copies/106 16S rRNA gene copies. The antibiotic treatment had no observable effect on antibiotic resistance gene or integron abundances.

Efficient removal of antibiotics in surface-flow constructed wetlands, with no observed impact on antibiotic resistance genes.

Recently, there have been growing concerns about pharmaceuticals including antibiotics as environmental contaminants. Antibiotics of concentrations commonly encountered in wastewater have been suggested to affect bacterial population dynamics and to promote dissemination of antibiotic resistance. Conventional wastewater treatment processes do not always adequately remove pharmaceuticals causing environmental dissemination of low levels of these compounds. Using constructed wetlands as an additional treatment step after sewage treatment plants have been proposed as a cheap alternative to increase reduction of wastewater contaminants, however this means that the natural microbial community of the wetlands becomes exposed to elevated levels of antibiotics. In this study, experimental surface-flow wetlands in Sweden were continuously exposed to antibiotics of concentrations commonly encountered in wastewater. The aim was to assess the antibiotic removal efficiency of constructed wetlands and to evaluate the impact of low levels of antibiotics on bacterial diversity, resistance development and expression in the wetland bacterial community. Antibiotic concentrations were measured using liquid chromatography-mass spectrometry and the effect on the bacterial diversity was assessed with 16S rRNA-based denaturing gradient gel electrophoresis. Real-time PCR was used to detect and quantify antibiotic resistance genes and integrons in the wetlands, during and after the exposure period. The results indicated that the antibiotic removal efficiency of constructed wetlands was comparable to conventional wastewater treatment schemes. Furthermore, short-term treatment of the constructed wetlands with environmentally relevant concentrations (i.e. 100-2000 ng×l(-1)) of antibiotics did not significantly affect resistance gene concentrations, suggesting that surface-flow constructed wetlands are well-suited for wastewater treatment purposes.

Occurrence and abundance of antibiotics and resistance genes in rivers, canal and near drug formulation facilities--a study in Pakistan.

Antibiotic resistance (AR) is a global phenomenon that has severe epidemiological ramifications world-wide. It has been suggested that antibiotics that have been discharged into the natural aquatic environments after usage or manufacture can promote the occurrence of antibiotic resistance genes (ARG). These environmental ARGs could serve as a reservoir and be horizontally transferred to human-associated bacteria and thus contribute to AR proliferation. The aim of this study was to investigate the anthropogenic load of antibiotics in Northern Pakistan and study the occurrence of ARGs in selected samples from this region. 19 sampling sites were selected; including six rivers, one dam, one canal, one sewage drain and four drug formulation facilities. Our results show that five of the rivers have antibiotic levels comparable to surface water measurements in unpolluted sites in Europe and the US. However, high levels of antibiotics could be detected in the downstream river in close vicinity of the 10 million city Lahore, 1100, 1700 and 2700 ng L(-1) for oxytetracycline, trimethoprim, and sulfamethoxazole respectively. Highest detected levels were at one of the drug formulation facilities, with the measured levels of 1100, 4100, 6200, 7300, 8000, 27,000, 28,000 and 49,000 ng L(-1) of erythromycin, lincomycin, ciprofloxacin, ofloxacin, levofloxacin, oxytetracycline, trimethoprim and sulfamethoxazole respectively. ARGs were also detected at the sites and the highest levels of ARGs detected, sulI and dfrA1, were directly associated with the antibiotics detected at the highest concentrations, sulfamethoxazole and trimethoprim. Highest levels of both antibiotics and ARGs were seen at a drug formulation facility, within an industrial estate with a low number of local residents and no hospitals in the vicinity, which indicates that the levels of ARGs at this site were associated with the environmental levels of antibiotics.

The development and application of a system for simultaneously determining anti-infectives and nasal decongestants using on-line solid-phase extraction and liquid chromatography-tandem mass spectrometry.

A method for the simultaneous analysis of antibiotics, antiviral and nasal decongestants in treated sewage effluent and surface water has been developed and validated. The method uses on-line solid phase extraction (SPE) of injected high-volume samples in conjunction with liquid chromatography-tandem mass spectrometry (LC-MS/MS). This method includes a range of antibiotics (Trimethoprim, Oxytetracycline, Ofloxacin, Norfloxacin, Ciprofloxacin, Azithromycin, Doxycycline, Sulfamethoxazole, Erythromycin and Clarithromycin), an antiviral (Oseltamivir) and nasal decongestants (Naphazoline, Oxymetazoline and Xylometazoline). The method's detection limits (MDLs) ranged from (0.2 ng L(-1)) to (3.1 ng L(-1)), based on a 1 mL extraction volume. Its intra-day precision was determined by performing nine runs with 200 ng L(-1) samples; the intra-day relative standard deviation (RSD) ranged from 1% to 19%. Inter-day precision was determined by analyzing samples in triplicate over the course of three days, yielding relative standard deviations ranging from <5% to <26%. The linearity (R(2)) for all compounds tested was >0.90. Spike relative recoveries ranged from 40% to 157% and 40% to 152% for STP effluent and surface water samples, respectively. Finally, the method was used to analyze real effluent and surface water.