Prevalence of active HCV infection among the blood donors of Khyber Pakhtunkwa and FATA region of Pakistan and evaluation of the screening tests for anti-HCV

Hepatitis C is a fatal liver disease caused by the hepatitis C virus. In this study, blood donors, from various districts of the KPK province and the federally administered tribal area (FATA) of Pakistan were tested for anti-HCV antibodies and HCV RNA by ICT (Immuno-chromatographic test), ELISA and RT-PCR. Out of the 7148 blood donors, 224 (3.13%) were positive for anti-HCV antibodies by ICT, 135 (1.89%) by ELISA while 118 (1.65%) blood donors had active HCV infection as detected by RT-PCR. We suggest that ELISA should be used for anti-HCV screening in public sector hospitals and health care units

Molecular characterization of ochratoxin a producing indigenous Aspergillus strains from poultry feed in Pakistan

Ochratoxin A (OTA) is nephrocarcinogenic and immunosuppressive toxin and OTA producing molds contaminate the food crops. Isolation and identification of ochratoxin producing fungi was carried out from poultry feed samples (n=120) followed by preliminary confirmation through macroscopic and microscopic characteristics. Purified fungal isolates identified as Aspergillus 1842(91.68%) followed by Penicillium 91 (4.53%), Mucor 52 (2.58), Alternaria 7 (0.35%), Cladosporium 6 (0.29%), Fusarium 4 (0.199%) and unidentified (07). OTA production was confirmed through thin layer chromatography (TLC) followed by high performance liquid chromatography (HPLC). Only 41 isolates (2.22%) out of 1842 Aspergillus isolates were able to produce toxin. At genetic level, characterization was performed through polymerase chain reaction (PCR) using species specific gene primers. From 41 isolates 27, 9 and 5 were characterized as Aspergillus terreus, Aspergillus parasiticus, and Aspergillus ochraceus, respectively. Physical and chemical factors were optimized for OTA production. Under the effect of 37 °C temperature and 7.5 pH of Sabouraud dextrose broth (SDB) medium, higher toxin (969.45±.03 μg/mL) production was observed from ASPO-6 isolate. ASPO-4 isolate produce higher toxin amount in SDB medium with supplementation of maize 5%, wheat 1% and rice 3%. OTA stability was determined by adjusting standard concentration of 100 μg/mL in organic solvents (chloroform, acetonitrile and methanol) and organic solids. Least percentage log reduction in OTA concentration and stability of OTA was observed in opaque vials with chloroform and sucrose and transparent vials with sucrose after 6 months. OTA can be used as indigenous standard for identification of OTA from field samples

Characterization of beta-galactosidase and enzymatic synthesis of prebiotic galacto-oligosaccharides

Galaktooligosaccharide (GOS) sind unverdauliche Kohlenhydrate, welche durch Transgalaktosylierung von Laktose mithilfe des Enzyms ß-Galaktosidase hergestellt werden. GOS wirken als Präbiotika positiv auf die Darmmikrobiota, indem sie das Wachstum von probiotischen Bakterien, wie Laktobazillen und Bifidobakterien, fördern und jenes von Krankheitserregern unterdrücken. Deshalb wird diesem Forschungsbereich in den letzten Jahren besondere Aufmerksamkeit gewidmet. Die glykosidische Bindung zwischen den Monosaccharid-Resten der produzierten GOS hängt stark von dem Organismus ab, aus welchem die entsprechenden lacLM Gene isoliert wurden, sowie den Transgalaktosylierungs-Bedingungen und bestimmt die präbiotische Effizienz. In der vorliegenden Arbeit wurden rekombinante ß-Galaktosidasen aus probiotischen Lactobacillus plantarum und L. sakei biochemisch charakterisiert. Diese beiden Enzyme sind Heterodimere aus der Familie der Glykosid Hydrolasen GH2 und ihr Temperatur- bzw. pH-Optimum ist weitgehend zur Produktion von GOS aus Laktose geeignet. Die ß-Galaktosidasen aus Lactobacillus plantarum und L. sakei bilden bevorzugt GOS mit ß-(1¿6) und ß-(1¿3) Bindungen, wobei hauptsächlich ß-D-Galp-(1¿3)-D-Glc, ß-D-Galp-(1¿6)-D-Glc, ß-D-Galp-(1¿3)-D-Gal, ß-D-Galp-(1¿6)-D-Gal, ß-D-Galp-(1¿6)-D-Lac und ß-D-Galp-(1¿3)-D-Lac produziert werden. Der Ausbeute an GOS beträgt ca. 41 % (w/w) vom Gesamtzucker. Zusammenfassend ist die Charakterisierung verschiedener ß-Galaktosidasen und ihrer Transgalactosylierungs-Aktivitäten, besonders aus probiotischen Bakterien, von großer Bedeutung.Galacto-oligosaccharides (GOS) are non-digestible carbohydrates which meet the criteria of “prebiotics” and therefore have attracted increasing attention in last few years. These oligosaccharides are produced by transgalactosylation of lactose, catalyzed by -galactosidase, and have been found capable of modulating the colonic microflora by stimulating the growth of beneficial bacteria (“probiotics”) especially bifidobacteria and lactobacilli and suppressing the activities of pathogens. Prebiotic efficiency of GOS is influenced by the glycosidic linkage between monosaccharide residues which is strongly dependent on -galactosidase source and conditions of transgalactosylation. The recombinant -galactosidases from probiotic Lactobacillus plantarum and Lactobacillus sakei have been biochemically characterized in detail for various parameters. Both enzymes are heterodimeric -galactosidases encoded by lacLM genes, belonging to glycoside hydrolase family GH2. Temperature and pH requirements of the enzymes are quite suitable for the bioconversion of lactose for GOS production. These -galactosidases have high transgalactosylation activities with special preference to form -(16) and -(13)-linked GOS products during lactose conversion. The main components of GOS mixtures produced by these -galactosidases are -D-Galp-(13)-D-Glc, -D-Galp-(16)-D-Glc, -D-Galp-(13)-D-Gal, -D-Galp-(16)-D-Gal, -D-Galp-(16)-D-Lac and -D-Galp-(13)-D-Lac. The yield of GOS is 41% (w/w) of total sugars. In conclusion, it is of great interest to characterize different -galactosidases and to investigate their transgalactosylation activities, from different sources, preferably from probiotic bacteria.Sanaullah IqbalAbweichender Titel laut Übersetzung der Verfasserin/des VerfassersZsfassung in dt. SpracheWien, Univ. für Bodenkultur, Diss., 2011OeBB(VLID)193026

Effect of camel milk lactoferrin against carbon tetrachloride induced hepatic toxicity in Sprague Dawley rats

Background:Chronic Hepatitis and mortality due to liver cirrhosis is common in Pakistan. Camel milk Lactoferrin has antiviral, anti-inflammatory properties. Nutraceutical foods like camel milk have many uses. Camel milk lactoferrin might be used for the cure of hepatic fibrosis induced by CCI4 in Sprague Dawley rats.Methods: Seventy-five (75) male Sprague–Dawley rats were purchased from National Institute of Health Islamabad kept in animal house of UVAS Lahore and randomly divided into 5 groups under completely randomize design. In Each group carbon tetrachloride (CC14) was subcutaneously injected with a mixture of 40% CCI4 (a mixture of pure CC14 and sterile olive oil) at 200 uL/100g body weight as single dose, 48 hrs before the starting of treatment or 0 day. After 48 hrs. Rats were considered hepatic injured except the (+ve) control group. Among all groups, four groups –ve control, 30mg/kg/b.wt, 60mg/kg/b.wt and 90mg/kg/b.wt were supplied with standard diet plus Lactoferrin (different concentrations and doses) orally, while control group were provided only standard diet throughout the efficacy study (30 days).Results: The study was conducted in two phases, first one was on the isolation and purification of camel milk lactoferrin and the second one was based on efficacy study of lactoferrin in Sprague Dawley rats against the hepatic toxicity induced by carbon tetrachlorideConclusion: The present study on CCl4 induced hepatotoxicity in Sprague Dawley rats clarifies that camel milk lactoferrin induced significant improvement in serum level of ALP, AST, AST, bilirubin, serum urea and serum Creatinine within the duration of 4 weeks treatment.Keywords: Carbon tetrachloride; Complete randomized design; Aspartate aminotransferas

System and Reforms of Higher Education in Pakistan

Abstract Since the creation of Pakistan, higher education system has been a matter of great concern among various circles in Pakistan, especially among the educationists, Government, policy makers and general public (Isani, 2001). Ranging from socio-economic system to religio-cultural lives of people (ranking in human development, 2001), the issue of higher education has several dimensions (Isani, 2001). In 1974, to address various challenges in higher education, the Government of Pakistan established University Grant Commission (UGC) (GOP, 1974). The main aim of UGC was to maintain standard of education and establish a uniform policy all over the country (Government of Pakistan, 1973, 1974. However, having no real financial powers, the UGC remained an ineffective institution (UGC, 2000). Resultantly, the standard of higher education in Pakistan declined considerably (GOP, 1976). Therefore, in 2002, the government established Higher Education Commission (HEC). HEC paved the way to revitalize invigorate, support, legalize, standardize, sponsor and made functional the endeavors undertaken by the various respective intuitions quiescent culture into mobile and active gestures (World Bank, 1994). As the title indicates, this paper is an attempt to analyze the reforms in higher education system in Pakistan. Key terms: Higher Education Commission, brain drain, E-Learning, Education Polices Quality Education. Introduction Pakistan's educational history stretches over the span of 64 long years, many efforts and endeavors remained under progress during this span, but most of such efforts didn't come out successful. Political set up remained the least, army incursion and topple downs made the democratic regimes squeeze not beyond 35 years. The agenda of reforms took a long way right from 1948 to 1985, three full-fledged constitutions promulgate during the years The reforms agenda did not end it did include the collection of half hearted, terminated and abandoned efforts. There are many reports of commissions, panels, committees, task forces which had never been attended which comprises the Commissions of on Agriculture (1987), Commission of Local Government (1983), and Commission on Education (1959). There remain certain policies and plans which had long term effects to yield provided these have been attended to in the manner it was desired like education policies of 1970of , 1972of , 1979of , 199

Nutraceutical, Anti-Inflammatory, and Immune Modulatory Effects of β-Glucan Isolated from Yeast

β-Glucan is a dietary fibre, found in many natural sources, and controls chronic metabolic diseases effectively. However, β-glucan from the yeast has rarely been investigated. Objectively, conditions were optimized to isolate β-glucan from the yeast (max. 66% yield); those optimized conditions included 1.0 M NaOH, pH 7.0, and 90°C. The purity and identity of the isolated β-glucan were characterized through FT-IR, SEM, DSC, and physicofunctional properties. The obtained results from DSC revealed highly stable β-glucan (m.p., 125°C) with antioxidant activity (TAC value 0.240 ± 0.0021 µg/mg, H2O2 scavenging 38%), which has promising bile acid binding 40.463% and glucose control (in vitro). In line with these results, we evaluated the in vivo anti-inflammatory potential, that is, myeloperoxidase activity and reduction in MDA and NO; protective effect on proteins and keeping viscosity within normal range exhibited improvement. Also, the in vivo cholesterol binding and reduction in the skin thickness by β-glucan were highly encouraging. Finally, our results confirmed that yeast β-glucan is effective against some of the inflammatory and oxidative stress markers studied in this investigation. In general, the effect of 4%  β-glucan was more noticeable versus 2%  β-glucan. Therefore, our results support the utilization of β-glucan as a novel, economically cheap, and functional food ingredient

Exploring the Antibiotic Production Potential of Heterotrophic Bacterial Communities Isolated from the Marine Sponges Crateromorpha meyeri, Pseudaxinella reticulata, Farrea similaris, and Caulophacus arcticus through Synergistic Metabolomic and Genomic Analyses

The discovery of novel secondary metabolites is actively being pursued in new ecosystems. Sponge-associated bacteria have been in the limelight in recent years on account of their ability to produce bioactive compounds. In this study, heterotrophic bacteria associated with four sponge species were isolated, taxonomically identified, and subjected to screening for the production of bioactive entities against a panel of nine microorganisms, including Gram-positive and negative bacteria, as well as yeast and fungi. Of the 105 isolated strains, 66% were represented by Proteobacteria, 16% by Bacteriodetes, 7% by Actinobacteria, and 11% by Firmicutes. Bioactivity screening revealed that 40% of the total isolated strains showed antimicrobial activity against one or more of the target microorganisms tested. Further, active extracts from selective species were narrowed down by bioassay-guided fractionation and subsequently identified by HR-ESI-MS analyses to locate the active peaks. Presumably responsible compounds for the observed bioactivities were identified as pentadecenoic acid, oleic acid, and palmitoleic acid. One isolate, Qipengyuania pacifica NZ-96T, based on 16S rRNA novelty, was subjected to comparative metabolic reconstruction analysis with its closest phylogenetic neighbors, revealing 79 unique functional roles in the novel isolate. In addition, genome mining of Qipengyuania pacifica NZ-96T revealed three biosynthetic gene clusters responsible for the biosynthesis of terpene, beta lactone, lasso peptide, and hserlactone secondary metabolites. Our results demonstrate the ability to target the sponge microbiome as a potential source of novel microbial life with biotechnological potential

Influence of Yeast β-Glucan on Cookies Sensory Characteristics and Bioactivities

β-Glucan is biologically active polysaccharide, ubiquitously found in many grains, bacteria, and fungi and much yeast. The aim of this study was to determine the effect of substituting wheat flour by 1, 2, and 4% yeast isolated β-glucan in cookies on the sensory acceptance, antioxidants, oxidative stability, and quality evaluation which were investigated. According to the results, cookies supplemented at 2% yeast β-glucan were proved satisfactory on sensory quality perspective. During the storage study it was found that cookies made with 2 and 4% β-glucan have effectively (p>0.05) kept the peroxide value (PV) within acceptable range, demonstrating the promising role of β-glucan in deterring oxidative. It is further noted that 2 or 4% β-glucan incorporated cookies assimilated highest absorption spectra, suggesting the retardation in freshness losses, with having minimum microbial loads, showing microbiological safety. β-Glucan fortification in foods is technologically and economically feasible, suggesting that a significant prospect of β-glucan as low-cost food ingredient in formulating cookies at 2% offers exciting new use of β-glucan of yeast origin

Nutraceutical, Anti-Inflammatory, and Immune Modulatory Effects of β-Glucan Isolated from Yeast

β-Glucan is a dietary fibre, found in many natural sources, and controls chronic metabolic diseases effectively. However, β-glucan from the yeast has rarely been investigated. Objectively, conditions were optimized to isolate β-glucan from the yeast (max. 66% yield); those optimized conditions included 1.0 M NaOH, pH 7.0, and 90°C. The purity and identity of the isolated β-glucan were characterized through FT-IR, SEM, DSC, and physicofunctional properties. The obtained results from DSC revealed highly stable β-glucan (m.p., 125°C) with antioxidant activity (TAC value 0.240 ± 0.0021 µg/mg, H2O2 scavenging 38%), which has promising bile acid binding 40.463% and glucose control (in vitro). In line with these results, we evaluated the in vivo anti-inflammatory potential, that is, myeloperoxidase activity and reduction in MDA and NO; protective effect on proteins and keeping viscosity within normal range exhibited improvement. Also, the in vivo cholesterol binding and reduction in the skin thickness by β-glucan were highly encouraging. Finally, our results confirmed that yeast β-glucan is effective against some of the inflammatory and oxidative stress markers studied in this investigation. In general, the effect of 4%  β-glucan was more noticeable versus 2%  β-glucan. Therefore, our results support the utilization of β-glucan as a novel, economically cheap, and functional food ingredient

Impact of Novel Prebiotic Galacto-Oligosaccharides on Various Biomarkers of Colorectal Cancer in Wister Rats

Colorectal cancer (CRC) is one of the leading causes of cancer deaths around the globe. Bioactive food ingredients such as prebiotics have protective potential in colon cancer. Data on galacto-oligosaccharides (GalOS) against CRC are very limited and GalOS used in this study have β-1,6 and β-1,3 as major glycosidic linkages and, to our best knowledge, were never used before against any cancer treatment. This study aims to investigate the protective role of novel GalOS against various biomarkers of CRC including aberrant crypt foci (ACF), bacterial enzymes and short chain fatty acids (SCFA) in a rodent model induced with 1,2-dimethylhydrazine dihydrochloride (DMH). Inulin group was taken as positive control in present study to compare novel GalOS protective effects. GalOS doses of 76–151 mg and inulin doses of 114 mg were given to different groups treated with DMH. Results showed that ACF formation was significantly (p ≤ 0.05) less in high dose GalOS group (27.3%). GalOS also had protective effects against DMH-induced body weight loss and showed higher level of cecal and fecal SCFA (acetate, propionate and butyrate). High doses of GalOS also resulted in significant (p ≤ 0.05) reduction of bacterial enzymatic activities. Increased populations of beneficial bacteria (bifidobacteria and lactobacilli) and decreased concentrations of harmful bacteria were observed in all prebiotics treatment groups. It can be concluded that novel GalOS exhibit robust protective activity against ACF formation in vivo