Mechanistic Insights into Vancomycin Resistance in Clostridioides difficile

C. difficile is a nosocomial, opportunistic pathogen that has become more hypervirulent over the last decade. Vancomycin is currently a frontline antibiotic for the treatment of C. difficile infections. Yet, strains of C. difficile are becoming non-susceptible to vancomycin. Emergence of the vancomycin non-susceptible phenotype has led to various questions including: What resistance elements are present that might contribute to the phenotype? Are these elements located on the chromosome or a plasmid? Where did these elements originate from? To address these questions, whole genome sequence analysis was performed to survey genes that may be involved in the vancomycin non-susceptible phenotype. Bioinformatic analysis was used to answer whether putative vancomycin-resistance genes were chromosomal or plasmid based. Culture-dependent methods were used to detect enterococci from stools of patients from Texas and Kenya infected with C. difficile and this was supplemented with patient data to describe co-colonization in the gut and its effects on patients. The sequencing analysis revealed the presence of a putative van gene cluster in all isolates tested. The presence of vancomycin-resistance elements varied among the isolates and included: vanRS, vanW, vanH, and vanZ. In silico analysis of sequences suggested that the putative vancomycin-resistance genes were chromosomal. A high prevalence of enterococci and vancomycin-resistant enterococci (VRE) was observed in the stools of C. difficile patients from both Kenya and Texas. In addition, the presence of enterococci increased the duration of CDI symptoms and this might also promote selection of non-spore forming C. difficile isolates. Finally, high proportions of VRE and non-susceptible C. difficile isolates were found in the same stools, suggesting a potential source of resistance elements. Although rigorous functional analysis is needed to confirm the specific genes responsible for vancomycin non-susceptibility in C. difficile, the presence of vancomycin-resistance genes underlines a promising avenue for further experimentation. Knowing that the genes are likely chromosomal will help in downstream characterization of these genes. Given the correlation between vancomycin-resistant enterococci and the vancomycin non-susceptibility phenotype in C. difficile, and the shared epidemiology between the two bacteria, it is plausible that these resistance elements may have originated from Enterococci spp, however, further genetic analysis is needed to confirm this observation. Overall, this study has shed some light on the mechanism of vancomycin non-susceptibility in C. difficile and has highlighted avenues for further exploration. Moreover, the relationship between Enterococci spp. and C. difficile during infection has become clearer and might enhance future treatment strategie

Optimization and validation of high performance liquid chromatography-ultra violet method for quantitation of metoprolol in rabbit plasma: application to pharmacokinetic studies

Purpose: To develop a sensitive, simple and validated high performance liquid chromatography (HPLC) analytical method for the determination of metoprolol tartrate in rabbit plasma.Methods: Mobile phase of methanol and 50 mM ammonium dihydrogen phosphate solution (50:50) at pH 3.05 was used for separation of metoprolol on BDS hypersil C18 column at a wavelength of 223 nm. Flow rate and retention time were 0.6 mL/min and 7.4 min, respectively. For pharmacokinetic study, rabbits were given an oral dose of 8 mg/kg of metoprolol in solution form. Blood samples were taken from jugular vein of the rabbits after drug administration and analysed by HPLC.Results: Separation of metoprolol was not interfered with other components in plasma. The calibration curve was linear in the range of 25 - 1000 ng/mL (r2 = 0.997). Lower limits of detection (LLOD) and quantitation (LLOQ) were 8.87 and 25 ng/mL, respectively. Relative standard deviation (RSD) of intraday and inter-day precision was < 14.27 and 7.61 %, respectively. Relative error of accuracy was between 4.85 and 14.37 %. Maximum plasma concentration (Cmax), time to maximum plasma concentration (Tmax) and half-life (t½) after metoprolol oral administration in rabbits were 186.29 ng/mL, 0.50 h and 2.27 h, respectively.Conclusion: A simple, accurate and precise HPLC-UV method for metoprolol determination in rabbit plasma has been successfully developed and applied to a pharmacokinetic study.Keywords: HPLC-UV, Metoprolol, Pharmacokinetics, Rabbit plasma, Liquid-liquid extraction, Validatio

A Focus Group Discussion on Creation of Knowledge Economy in Punjab, Pakistan

World is moving from traditional economy to modern economy with more focus on knowledge. The creation of new knowledge is mandatory for developing any country economically. Institutions play important role in this transition. Especially educational institutions are momentous to bring change in existing system and are a major source to create new knowledge. Inductive method is used to understand the state of knowledge creation through educational institutions in Pakistan. Focus group discussions were arranged to collect the data. It is found that the educational institutions of Pakistan are slowly moving towards creations of knowledge economy. Improvements are needed to reform the educational system to create knowledge economy

A Focus Group Discussion on Creation of Knowledge Economy in Punjab, Pakistan

World is moving from traditional economy to modern economy with more focus on knowledge. The creation of new knowledge is mandatory for developing any country economically. Institutions play important role in this transition. Especially educational institutions are momentous to bring change in existing system and are a major source to create new knowledge. Inductive method is used to understand the state of knowledge creation through educational institutions in Pakistan. Focus group discussions were arranged to collect the data. It is found that the educational institutions of Pakistan are slowly moving towards creations of knowledge economy. Improvements are needed to reform the educational system to create knowledge economy

A Qualitative Analysis of Social Determinants of Entrepreneurship Success (Thematic Analysis)

This study has been conducted to explore the important social determinants of entrepreneurship success. Families’ involvement is crucial for entrepreneurship success. The myth is prevailed in our society as successful life is to be opened as to secure a position in public or private sector. Social determinants are not discussed in detail before. To know what social determinants are important to equip success in business? Qualitative exploratory case study method (Yin. K, 2014) was used to explore social determinants in detail. Field notes were taken on note book with pencil and interviews were recorded personally by the researcher himself and these field notes and data were transcribed to document the important patterns and themes. Thematic analysis was applied to the data for analysis purpose. Findings of the study depicted that family business involve in entrepreneurial intention, role model provide supportive material, family financial help, prior business experience, entrepreneurship knowledge and understanding, were the important determinants of entrepreneurship success.At the end of the paper it was suggested that entrepreneurship training must be provided by the government institution and universities must induct degree program in entrepreneurship. Seminars and conferences on entrepreneurship awareness must be organized by nongovernmental organizations with the collaboration of government agencies and institutions

Fabrication and Evaluation of Rosuvastatin Calcium Fast- Disintegrating Tablets Using β-Cyclodextrin and Superdisintegrants

Purpose: To formulate fast-disintegrating tablets (FDT) of rosuvastatin calcium (RST) using β- cyclodextrin (CD) and different superdisintegrants to enhance their solubility.Methods: A total of 15 FDT formulations of RST were prepared using three different techniques. The FDTs were evaluated for micromeritic properties, as well as by Fourier transform infrared spectroscopy (FTIR), thermal analysis, disintegration time (DT), dissolution rate, powder x-ray diffraction (XRDP), scanning electron microscopy (SEM) and stability studies.Results: XRDP showed that RST was changed from crystalline to amorphous form. SEM images revealed the presence of small microscopic pores that enhanced water penetration and provided rapid dissolution rate compared with the pure drug. There was maximum release of drug (99 %) from F4 formulation containing solid dispersion of RST, CD and superdisintegrants. DT and wetting time were 25 s (p = 0.032) and 33 s (p = 0.023), respectively, for F4 formulation. In vitro dispersion time was also lowest for F4 at 23 s (p = 0.023). FTIR and DSC studies also confirmed complex formation of drug with CD and superdisintegrants.Conclusion: FDT is a suitable strategy to enhance the dissolution rate of RST and thus is an effective tool to improve bioavailability of poorly water soluble drugs.Keywords: Solubility, β-cyclodextrin, Kyron, Polymer, Rosuvastatin, Fast-disintegrating tablet

Urban-Rural Differentials of Health and Educational Inequality in District of Faisalabad: A Social Analysis

This study aims to find out the relationship between socioeconomic determinants of rural-urban social inequity specifically with regard to health and educational facilities and the public perception of this inequality in Faisalabad District, Punjab, Pakistan. The data were collected through a survey by using interviewing schedules. The multistage sampling technique was used to collect data. At the first stage two Tehsils of district Faisalabad were selected randomly, while at the second stage four union councils, two from each Tehsil (one urban and one rural) were randomly selected. A specimen of 240 respondents, 120 from each Tehsil (60 from urban and 60 from country regions) were chosen arbitrarily from the selected regions. Data were analyzed by statistical analysis using SPSS software. Results of study revealed that peoples living in urban areas had better literacy level than rural peoples. T-statistics also revealed sharp contrast between urban and rural population regarding availability of basic education and health facilities. The study also revealed that urban people were more satisfied about available health and educational infrastructure. The study concludes that rural peoples are badly facing the impacts of social disparity due to lack of health and educational facilities as compared to urban peoples

Impedimentos enfrentados por instituições educacionais na criação de economia do conhecimento: um estudo de caso de Punjab Paquistão

Knowledge has become a powerful tool for the development of economy. The important role played by educational institutions in the production of knowledge economy cannot be denied. Pakistan is a developing country and has limited resources to deal with the new challenges of economy, so the research was designed to find out the problem faced by educational institutions in the process of creation of knowledge economy. The research was conducted into two parts. In the first part, qualitative approach is used to develop variables. These variables are further analyzed by using quantitative inquiry. Collected data is analyzed by using the SPSS. Descriptive statistics are applied to data to get the results. Results guide us that educational institutions are facing problems in providing access to knowledge, rich infrastructure, funds, incentives, research and development, developed human capital, and developing link between industry and universities. These problems are impeding the process of knowledge creation. Knowledge creation can only be enhanced by dealing the above mentioned problem.El conocimiento se ha convertido en una herramienta eficaz para el desarrollo de la economía. El rol desempeñado por las instituciones educativas en la producción de conocimiento de la economía no puede ser denied. El Pakistán es un país de transición y tiene recursos limitados para satisfacer los nuevos desafíos de la economía, ya que la investigación está diseñada para resolver el problema de las instituciones educativas en el proceso de creación de conocimiento de la economía. La investigación se llevó en dos partes. En el primer paso, se utilizó un método de aproximación para desarrollar variables. Estas variables se analizan mediante la utilización de cuantitativos cuantitativos. Se ha recopilado la fecha con el SPSS. La descripción estática se aplica a la fecha para obtener los resultados. En el caso de que se produzca un cambio en la calidad de la información, se debe tener en cuenta que, Estos problemas impiden el proceso de creación de conocimiento. La concepción de Knowledge Base sólo se puede mejorar al tratar el problema anteriormente mencionado.O conhecimento tornou-se uma ferramenta poderosa para o desenvolvimento da economia. O importante papel desempenhado pelas instituições educacionais na produção da economia do conhecimento não pode ser negado. O Paquistão é um país em desenvolvimento e tem recursos limitados para lidar com os novos desafios da economia, então a pesquisa foi projetada para encontrar o problema enfrentado pelas instituições educacionais no processo de criação da economia do conhecimento. A pesquisa foi realizada em duas partes. Na primeira parte, abordagem qualitativa é usada para desenvolver variáveis. Essas variáveis são analisadas posteriormente usando a pesquisa quantitativa. Os dados coletados são analisados usando o SPSS. Estatísticas descritivas são aplicadas para obter os resultados. Os resultados nos orientam que as instituições de ensino estão enfrentando problemas para fornecer acesso ao conhecimento, rica infraestrutura, fundos, incentivos, pesquisa e desenvolvimento, desenvolvimento de capital humano e desenvolvimento do vínculo entre a indústria e as universidades. Esses problemas estão impedindo o processo de criação do conhecimento. A criação de conhecimento pode ser melhorada ao lidar com o problema mencionado acima

Reformas educacionais necessárias para a criação da economia do conhecimento: um estudo de caso de Punjab Paquistão

In current era the economic growth depends on the creation of knowledge economy and production and distribution of knowledge has become the critical element of economic development. Educational institution play key role in creation and distribution of knowledge. So, the prime objective of the research was to find out the reforms which can help educational institutions to create knowledge economy. Both the qualitative and quantitative approaches were used to collect the data. Teachers, students, and administrators were selected from the both private and public educational institutions of the Punjab, Pakistan. FGDs were arranged to collect the qualitative data and the quantitative data was gathered by survey method. The collected information was analyzed by using descriptive statistics techniques. Access to knowledge, rich infrastructure, funds, incentives, research and development, human capital development and collaboration with the industry are the suggested ways by the respondents to increase the creation of knowledge economy through the educational institutions of Pakistan.En la era actual, el crecimiento económico depende de la creación de la economía del conocimiento y la producción y distribución del conocimiento se ha convertido en el elemento crítico del desarrollo económico. Las instituciones educativas desempeñan un papel clave en la creación y distribución del conocimiento. Entonces, el objetivo principal de la investigación fue descubrir las reformas que pueden ayudar a las instituciones educativas a crear una economía del conocimiento. Se utilizaron los enfoques tanto cualitativos como cuantitativos para recopilar los datos. Los maestros, estudiantes y administradores fueron seleccionados de las instituciones educativas privadas y públicas del Punjab, Pakistán. Se dispusieron FGD para recopilar los datos cualitativos y los datos cuantitativos se recopilaron por el método de encuesta. La información recolectada fue analizada utilizando técnicas de estadística descriptiva. El acceso a los conocimientos, una infraestructura rica, fondos, incentivos, investigación y desarrollo, desarrollo de capital humano y colaboración con la industria son las formas sugeridas por los encuestados para aumentar la creación de la economía del conocimiento a través de las instituciones educativas de Pakistán.Na era atual, o crescimento econômico depende da criação da economia do conhecimento e a produção e distribuição do conhecimento tornou-se o elemento crítico do desenvolvimento econômico. A instituição educacional desempenha papel fundamental na criação e distribuição de conhecimento. Assim, o objetivo principal da pesquisa foi descobrir as reformas que podem ajudar as instituições educacionais a criar economia do conhecimento. Ambas as abordagens, qualitativa e quantitativa, foram usadas para coletar os dados. Professores, estudantes e administradores foram selecionados entre instituições educacionais privadas e públicas do Punjab, no Paquistão. FGDs foram organizados para coletar os dados qualitativos e os dados quantitativos foram coletados pelo método de pesquisa. As informações coletadas foram analisadas por meio de técnicas de estatística descritiva. O acesso ao conhecimento, rica infra-estrutura, fundos, incentivos, pesquisa e desenvolvimento, desenvolvimento do capital humano e colaboração com a indústria são as formas sugeridas pelos entrevistados para aumentar a criação da economia do conhecimento através das instituições educacionais do Paquistão

Fabrication of Demineralized Bone Matrix/Polycaprolactone Composites Using Large Area Projection Sintering (LAPS)

Cadaveric decellularized bone tissue is utilized as an allograft in many musculoskeletal surgical procedures. Typically, the allograft acts as a scaffold to guide tissue regeneration with superior biocompatibility relative to synthetic scaffolds. Traditionally these scaffolds are machined into the required dimensions and shapes. However, the geometrical simplicity and, in some cases, limited dimensions of the donated tissue restrict the use of allograft scaffolds. This could be overcome by additive manufacturing using granulated bone that is both decellularized and demineralized. In this study, the large area projection sintering (LAPS) method is evaluated as a fabrication method to build porous structures composed of granulated cortical bone bound by polycaprolactone (PCL). This additive manufacturing method utilizes visible light to selectively cure the deposited material layer-by-layer to create 3D geometry. First, the spreading behavior of the composite mixtures is evaluated and the conditions to attain improved powder bed density to fabricate the test specimens are determined. The tensile strength of the LAPS fabricated samples in both dry and hydrated states are determined and compared to the demineralized cancellous bone allograft and the heat treated demineralized-bone/PCL mixture in mold. The results indicated that the projection sintered composites of 45–55 wt %. Demineralized bone matrix (DBM) particulates produced strength comparable to processed and demineralized cancellous bone