Molecular characterization of mycotoxin-producing Aspergillus parasiticus and sensitivity pattern to different disinfectants

Aims@#The study was aimed to isolate and characterize the mycotoxin-producing filamentous Aspergillus parasiticus from the feed samples. The sensitivity pattern of the isolates was assessed against different disinfectants.@*Methodology and results@#Fifty different feed samples were screened for A. parasiticus isolation. Isolates were subjected to macroscopic and microscopic characterization. Polymerase chain reaction was performed to confirm the isolates at the genomic level. Mycotoxin producing potential of the isolates was assessed by thin-layer chromatography (TLC). To quantify the toxins, high performance liquid (HPLC) was employed. The antifungal potential of disinfectants was determined by the well diffusion method followed by minimum inhibitory concentration (MIC) calculation. Out of twenty isolates of A. parasiticus, 11(55%) isolates were observed positive for toxin production. Three toxigenic isolates (AspP2, AspP4 and AspP8) were selected to evaluate their susceptibility against disinfectants by well diffusion method. AspP2 produced maximum (5.90 ng/mL) toxin, followed by AspP4 (3.11 ng/mL) and AspP8 (18.47 ng/mL). Terralin showed maximum fungicidal activity with 29.66 ± 8.08 mm zone of inhibition at 0.42 μg/mL MIC. Hypochlorite and Instru Star showed 99% disinfection with 30, 60 and 90 min contact time (6 mean log reduction) for all A. parasiticus isolates. Alpha Guard inhibited growth after 15 min contact time for all the isolates.@*Conclusion, significance and impact of study@#This study provides data indicating the contamination of feed samples with mycotoxin-producing A. parasiticus isolates and their sensitivity against commercially available disinfectants. Use of these disinfectants in appropriate concentrations and time could help prevent the contamination of food, feed and healthcare settings with the fungal species.

Antibacterial potential of indigenous medicinal plants against methicillin-resistant Staphylococcus aureus isolated from septic wounds

Aims@#This study was aimed to screen indigenous medicinal plants for their antibacterial potential against methicillin-resistant Staphylococcus aureus (MRSA).@*Methodology and results@#Three indigenous plants (Nigella sativa, Zingiber officinale and Calotropis procera) and thymoquinone were screened for antibacterial activity against MRSA, isolated from septic wounds of patients admitted to Mayo Hospital Lahore, Pakistan. Isolated bacteria were screened for methicillin and cefoxitin resistance by the Kirby-Bauer method, followed by mecA gene-specific polymerase chain reaction (PCR). Confirmed MRSA was processed for antibacterial activity of plant extracts and thymoquinone followed by cytotoxicity assay of plant extract having least minimum inhibitory concentration (MIC) value. Out of total samples (n=100), S. aureus (29%), MRSA (26%) and vancomycin-resistant S. aureus (VRSA) (21.7%) isolates were recovered based on morphology, biochemical profile and antibiotic susceptibility testing. Nigella sativa showed the highest antibacterial activity (10.06 ± 6.53 mm) against MRSA followed by Z. officinale (4.06 ± 3.72 mm) and C. procera (3.65 ± 3.33 mm) in comparison to standard thymoquinone (17.93 ± 10.14 mm). The least MIC value recorded was for Z. officinale at 36.89 ± 3.75 μg/mL. Zingiber officinale was the most effective antibacterial agent, followed by N. sativa and C. procera and non-toxic for eukaryotic cells at all tested concentrations (1500 μg/mL to 2.92 μg/mL).@*Conclusion, significance and impact of study@#It was concluded that Z. officinale may be used as an effective alternative for treating septic wound infection in local or topical preparations. As pathogenic S. aureus is becoming life-threatening among antibiotic-resistant bacteria and traditional plants are in used for centuries to treat septic wound infections.

Biofuel production potential of indigenous isolates of Scenedesmus sp. from lake water in Pakistan

Aims@#This paper presents the report on biodiesel and biogas production at a laboratory scale from Scenedesmus strain.@*Methodology and results@#Previously isolated and identified Scenedesmus were grown in 10 Liter flask using BG-11 media at 16 h light and 8 h dark cycle. Oven-dried biomass (20 g) from 16-day-old culture of Scenedesmus was finely grounded and subjected to lipids extraction by chloroform-methanol-NaCl mixture. Microalgal lipids (6 mL) were subjected to transesterification by using NaOH leading to the production of 5 mL biodiesel and 4 mL of glycerin. Biodiesel was rich in methyl esters of linoleic acid, phosphorothioc acid and dodecanoic acid, as shown by gas chromatography-mass spectrometry (GC-MS) analysis. Oven-dried microalgae (2 g) without lipid extraction and leftover biomass (2 g) after lipid extraction were subject to biogas production through anaerobic digestion. Biogas (34, 27 and 19 mL) were recorded respectively in oven-dried whole biomass; lipid extracted biomass and control over a period of 15 days of anaerobic digestion.@*Conclusion, significance and impact of study@#It was concluded that water bodies are rich in diverse algae, especially Scenedesmus sp., and this algae can be cultured to produce biodiesel and biogas. But the lipid accumulation potential of microalgae requires special treatment and lipid extraction methods are not up to the mark, which is a major bottleneck in biofuel production from microalgae.

A novel polyherbal formulation containing thymoquinone attenuates carbon tetrachloride-induced hepatorenal injury in a rat model

Objective: To evaluate a novel polyherbal formulation (BSVT) containing the standardized extracts from the leaves of Boerhavia diffusa, Solidago virgaurea, Vitex negundo, and thymoquinone in CCl

Augmented cytotoxic, mutagenic and genotoxic response triggered by carvedilol and celecoxib combinations

It is understood that drugs regardless of their order of administration can exhibit drug interactions. Established on the fact that treatment of hypertension may last for decades and prolong usage of multiple drug regimen may induce substantial pathophysiological changes. Hence, This study was designed to evaluate the possible synergistic toxic effects of anti-hypertensive (carvedilol), and anti-inflammatory drug (celecoxib) alone and in combinations. Well-established MTT assay, Single Cell Gel Electrophoresis (SCGE) and Ames assay were employed to evaluate the toxicity at cellular level. Results from MTT assay on Vero cell line revealed that drug combinations have more pronounced anti-proliferative activity with combine IC50 value of 13.7:47.8 µg/mL. Likewise, exposure of peripheral blood mononuclear cells with drug combinations revealed significant (P<0.05) DNA damage (Class 3) in a dose dependent manner at concentrations ≥ 0.78: 2.34 µg/mL. However, carvedilol and celecoxib were non mutagenic against either mutant strain (TA 100 and TA 98) and combinations have also shown mild to moderate mutagenic potential. Nevertheless, upon addition of metabolic activation enzyme, concentration <12.5:37.5 µg/plate exhibited significant (P<0.05) mutagenicity against both tester strains. In conclusion, this study provides additional genotoxicity and mutagenicity data that could be used in considering options for formulating regimens with reduced mutagenic potential

A consensus of key opinion leaders on the management of pre-diabetes in the Asia-Pacific region / Journal of the ASEAN Federation of Endocrine Societies

@#<p>The Asia-Pacific region carries a high disease burden, with over half of the global diabetic population residing in this region. Increasing evidence shows that without targeted intervention, the progression from impaired glucose tolerance (IGT) to type 2 diabetes occurs more frequently in Asians compared with Caucasians. Furthermore, IGT is independently associated with an increased risk of cardiovascular disease, and should be managed as early as possible. Because diabetes is now a major public health issue, strategies aimed at prevention and treatment are urgently required. Lifestyle modification, including weight loss, dietary changes and increased physical activity, play a major role in controlling the disease. Significant evidence also supports the effectiveness of a combination of lifestyle modification and pharmacologic therapy, such as metformin, in delaying the onset of diabetes. Although the importance of lifestyle interventions is well recognized throughout Asia, many countries do not have formal recommendations to guide the diagnosis and management of individuals at risk of progression to diabetes. At a recent regional meeting, experts from the Asian region convened to develop consensus recommendations to guide clinicians in the management of Asian patients with pre-diabetes. These consensus recommendations provide a clear and concise approach to the management of individuals with IGT based on the available evidence and current best clinical practice.</p>

Nigella sativa protects against isoproterenol-induced myocardial infarction by alleviating oxidative stress, biochemical alterations and histological damage

Objective To evaluate the cardioprotective effect of Nigella sativa L. (N. sativa) in isoproterenol-induced myocardial infarction (MI). Methods Groups were treated with different doses of ethanol extract of N. sativa (EENS) and N. sativa oil alone and along with enalapril for 28 days. MI was induced by subcutaneous administration of isoproterenol (85 mg/kg) in two consecutive doses. Levels of cardiac biomarkers and antioxidant enzymes such as creatine kinase–N-acetyl-L-cysteine, lactate dehydrogenase, aspartate aminotransferase, malondialdehyde, superoxide dismutase, reduced glutathione and catalase were evaluated along with gross histopathological examination. Results Isoproterenol (85 mg/kg) induced MI by causing the significant (P < 0.01) reduction in the activity of cardiac biomarkers (creatine kinase–N-acetyl-L-cysteine, lactate dehydrogenase, aspartate aminotransferase) and antioxidant markers (superoxide dismutase, catalase, glutathione) along with significant (P < 0.01) increase in the level of malondialdehyde. Furthermore, histopathological evaluation also confirmed the isoproterenol-induced MI. Pretreatment with EENS (800 mg/kg) and combination of EENS (800 mg/kg) with enalapril (1 mg/kg) significantly (P < 0.01) prevented the development of these alteration and restored activity of cardiac biomarkers as well as antioxidant markers almost near to normal levels. Histopathological evaluation of cardiac tissue further confirmed the restoration of biochemical activity. Conclusions Experimental findings thus indicate that EENS (800 mg/kg) demonstrated cardioprotective effect against isoproterenol-induced MI by restoring cardiac biomarkers and antioxidant status.

Comparative studies on rabbit plasma lipid profile fed with Silybum marianum oil, sunflower oil and vegetable ghee

Present work is aimed to compare the physicochemical characterization and biochemical effects of oil extracted from Silybum Marianum and Sunflower oil, collected from Peshawar [Pakistan]. To investigate the comparative effects on the body weight, organ weight and lipid profile, the crude oil of Silybum marianum, edible sunflower oil and vegetable ghee were given to three groups of rabbits under study. Percent proximate composition and food consumption of all rabbits were determined which showed no significant statistical variation. There is no data available about Silybum marianum oil on animal model in literature. This study clearly revealed that oil from Silybum marianum significantly reduces plasma cholesterol level in rabbits. A threefold higher Triglyceride levels was observed in vegetable ghee feeding groups compared with the sunflower and Silybum marianum oil feeding groups. The crude oil of Silybum marianum was found to be safe in rabbits compared with sunflower oil and vegetable ghee. The results of these studies revealed most valuable information and also support the refining and purification to convert this non-edible oil to edible oil

Differential antibrucella activity of bovine and murine macrophages.

Brucella abortus is an intracellular pathogen affecting macrophages. Macrophages release some antibrucella components such as lysozymes (LZ), reactive oxygen species (ROS) and reactive nitrite intermediates (RNI) which prevent intracellular survival of Brucella. The present study compared the antibrucella activity of bovine and murine macrophages following stimulation with B. abortus lipopolysaccharides. Our results revealed increased production of these antibrucella substances in murine macrophages as compared to bovine macrophages. The differential production of these antibrucella components explained the differential B. abortus killing ability of these species (bovine and mice) that was measured in terms of intramacrophagic survival of Brucellae in murine and bovine macrophages.

A consensus of key opinion leaders on the management of pre-diabetes in the Asia-Pacific Region / Journal of the ASEAN Federation of Endocrine Societies

@#The Asia-Pacific region carries a high disease burden, with over half of the global diabetic population residing in thisregion. Increasing evidence shows that without targeted intervention, the progression from impaired glucose tolerance(IGT) to type 2 diabetes occurs more frequently in Asians compared with Caucasians. Furthermore, IGT is independently associated with an increased risk of cardiovascular disease, and should be managed as early as possible. Because diabetes is now a major public health issue, strategies aimed at prevention and treatment areurgently required. Lifestyle modification, including weight loss, dietary changes and increased physical activity, play a major role in controlling the disease. Significant evidence also supports the effectiveness of a combination of lifestylemodification and pharmacologic therapy, such as metformin, in delaying the onset of diabetes. Although the importanceof lifestyle interventions is well recognized throughout Asia, many countries do not have formal recommendations to guide the diagnosis and management of individuals at risk of progression to diabetes. At a recent regional meeting,experts from the Asian region convened to develop consensus recommendations to guide clinicians in themanagement of Asian patientswith pre-diabetes. These consensus recommendations provide a clear and conciseapproach to the management of individuals with IGT based on the available evidence and current best clinical practice.

Synthesis and biological evaluation of a mutual prodrug of norfloxacin and fenbufen

Objectives: The study aimed to synthesize a mutual prodrug of norfloxacin and fenbufen with an objective of obtaining an effective and safer anti-inflammatory drug with useful antimicrobial actions

Methods: An amide-based mutual prodrug [NF-FN] was prepared following a single-step synthesis by condensing norfloxacin with fenbufen under appropriate laboratory conditions. Its structure was established on the basis of IR, NMR, Mass spectral data and elemental analysis. The prodrug [NF-FN] was evaluated for in-vitro antibacterial activity against two grampositive [Staphylococcus aureusand Bacillus subtilis] and two gram negative bacterial strains [Escherichia coli and Klebsiella pneumonia]. The in-vivo antiinflammatory activity and ulcerogenicity of the synthesized prodrug were investigated in Wistar albino rats at the doses of 10 and 30 mg/kg body weight, respectively

Results: The synthesized prodrug [NF-FN] showed very good activity against S. aureus and E. coli with MIC-6.25 mg/ mL, and good activity against B. subtilis and K. pneumonia with MIC-12.5 mg/mL. Its anti-inflammatory activity was found to be better than that of the parent drug fenbufen. It was also observed to less severe on gastric mucosa in comparison to reference drug, fenbufen

Conclusion: The prodrug showed promising results as anti-inflammatory agent however, its antibacterial action was found to be slightly weaker than the other parent drug norfloxacin

Optogenetics Applications for Treating Spinal Cord Injury

Cases of spinal cord injury (SCI) are increasing all over the world; and in USA alone, there are 273,000 patients, which not only leads to morbidity and mortality but also results in a great economic burden. Many approaches are being used at the pre-clinical and clinical level to treat SCI including therapeutic agents, surgical decompression, stem cell therapy etc. Recently, a new approach called optogenetics has emerged in which light sensitive proteins are used to switch neurons on and off, and this approach has great potential to be used as therapy due to its specificity and rapid response in milliseconds. Few animal studies have been performed so far in which the respiratory and bladder function of rats was restored through the use of optogenetics. On the basis of promising results obtained, in the future, this approach can prove to be a valuable tool to treat patients with SCI.

Stem Cell Research in Pakistan; Past, Present and Future

BACKGROUND AND OBJECTIVES: Stem cells have proved to have great therapeutic potential as stem cell treatment is replacing traditional ways of treatment in different disorders like cancer, aplastic anemia, stroke, heart disorders. The developed and developing countries are investing differently in this area of research so research output and clinical translation of research greatly vary among developed and developing countries. Present study was done to investigate the current status of stem cells research in Pakistan and ways to improve it. RESULTS: Many advanced countries (USA, UK and Canada etc.) are investing heavily in stem cell research and treatment. Different developing countries like Iran, Turkey and India are also following the developed countries and investing a lot in stem cells research. Pakistan is also making efforts in establishing this field to get desired benefits but unfortunately the progress is at very low pace. If Government plays an active role along with private sector, stem cell research in Pakistan can be boosted up. The numbers of publications from Pakistan are very less compared to developed and neighboring countries and Pakistan also has very less number of institutes working in this area of research. CONCLUSIONS: Stem cells research is at its initial stages in Pakistan and there is great need to bring Government, academia and industry together so they could make serious efforts to promote research in this very important field. This will help millions of patients suffering from incurable disorders and will also reduce economic loss.

Nosocomial infections and their control strategies

Nosocomial infections are also known as hospital-acquired/associated infections. National Healthcare Safety Network along with Centers for Disease Control for surveillance has classified nosocomial infection sites into 13 types with 50 infection sites, which are specific on the basis of biological and clinical criteria. The agents that are usually involved in hospital-acquired infections include Streptococcus spp., Acinetobacter spp., enterococci, Pseudomonas aeruginosa, coagulase-negative staphylococci, Staphylococcus aureus, Bacillus cereus, Legionella and Enterobacteriaceae family members, namely, Proteus mirablis, Klebsiella pneumonia, Escherichia coli, Serratia marcescens. Nosocomial pathogens can be transmitted through person to person, environment or contaminated water and food, infected individuals, contaminated healthcare personnel's skin or contact via shared items and surfaces. Mainly, multi-drug-resistant nosocomial organisms include methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, Pseudomonas aeruginosa and Klebsiella pneumonia, whereas Clostridium difficile shows natural resistance. Excessive and improper use of broad-spectrum antibiotics, especially in healthcare settings, is elevating nosocomial infections, which not only becomes a big health care problem but also causes great economic and production loss in the community. Nosocomial infections can be controlled by measuring and comparing the infection rates within healthcare settings and sticking to the best healthcare practices. Centers for Disease Control and Prevention provides the methodology for surveillance of nosocomial infections along with investigation of major outbreaks. By means of this surveillance, hospitals can devise a strategy comprising of infection control practices.

Viral hepatitis in Pakistan: Challenges and priorities

Hepatitis B and C are big health issues worldwide as more than 400 million people are suffering from chronic hepatitis B and C which result in more than 1.4 million deaths each year. According to a study done by Pakistan Medical Research Council in 2007-08, 7.6% Pakistani population suffered with hepatitis B and C, with around 4.8% with hepatitis C only. Government of Pakistan has taken different initiatives like vaccination, patient safety, blood screening, education and awareness about disease but still there is high prevalence of hepatitis in Pakistan. According to some studies injecting drug users have the highest prevalence of hepatitis B and C in the country. The follow-up studies and documentation of hepatitis patients was not very good which need to be improved. There is no recent large scale study on risk factors and prevalence of hepatitis B and C in Pakistan so it should be done on an urgent basis. If government set up regional laboratories for prevalence study and also a central institute for hepatitis research and treatment, the disease could be prevented in better and proper way. The treatment of hepatitis is very costly and a developing country like Pakistan cannot afford such high costs. Therefore more focus should be on preventive measures.

Quantification of total phenol, flavonoid content and pharmacognostical evaluation including HPTLC fingerprinting for the standardization of Piper nigrum Linn fruits

Objective: To carry out the physicochemical and phytochemical standardization with high performance thin layer chromatography fingerprinting of Piper nigrum L. (P. nigrum) fruits in order to ascertain the standard pharmacognostical parameters of this king of spices. Methods: Many standardization parameters like extractive values, total ash value, water soluble ash value and acid insoluble ash, moisture content, loss on drying and pH values of P. nigrum L. fruits were analyzed. The method of Harborne was adopted for the preliminary phytochemicals screening. Analysis of total phenolic and flavonoid contents, pesticides residues, aflatoxin and heavy metals were also performed. CAMAG-high performance thin layer chromatography system was used for fingerprinting of methanolic extract of P. nigrum L. fruits. Results: The results of phytochemicals testing indicated the presence of carbohydrates, phenolic compounds, flavonoids, alkaloids, proteins, saponins, lipids, sterols and tannins in various solvent extracts. Total phenolic and flavonoid contents in methanolic extract were found to be 1.728 1 mg/g and 1.087 μg/g, respectively. Heavy metals concentrations were found to be within standard limits. Aflatoxins and pesticides residues were absent. Conclusions: The outcome of this study might prove beneficial in herbal industries for identification, purification and standardization of P. nigrum L. fruits.

Preparation and in vitro evaluation of Nystatin micro emulsion based gel

Nystatin is a polyene antimycotic obtained from Streptomyces noursei used in the treatment of topical and transdermal fungal infection. Nystatin is nearly insoluble in water [<0.1] and it is amphoteric in nature. The aim of the present study was to design and develop Nystatin micro emulsion based gel for efficient delivery of drug to the skin by water titration method. The Pseudoternary phase diagrams 1:2, 1:1 and 2:1 were constructed by water titration method. Micro emulsion based gel was prepared by using oleic acid, Tween 20, propylene glycol as an oil phase, surfactant and cosurfactant respectively. Cabopol 940 was used as a gelling agent. In vitro evaluation of micro emulsion based gel was done for pH, Viscosity, spreadability and droplet size. Micro emulsion based gel showed greater antifungal activity against Candida albicansas compared to control formulations. In vitro drug release studies were conducted for micro emulsion based gel and control formulation using Franz diffusion cell. Drug penetration through synthetic skin followed Zero order model as the values for R[2] higher in case of zero order equation. The optimized micro emulsion based gel was found to be stable and showed no physical changes when exposed to different temperatures for a period of 4 week. The results indicated that the micro emulsion based gel system studied would be a promising tool for enhancing the percutaneous delivery of Nystatin

Evaluation of cytotoxicity and antiviral activity of ivermectin against newcastle disease virus

Cytotoxic and antiviral potential of ivermectin and ribavirin was evaluated. Cytotoxicity was checked on chick primary fibroblast cell line through MTT assay. Antiviral potential was determined against Newcastle disease virus on 9- day old chicken embryos. Six different concentrations [200, 100, 50, 25, 12.5 and 6.25[micro]g/mL] of both the drugs were evaluated. The 100[micro]g/mL concentration of ivermectin and higher were cytotoxic. The 25[micro]g/mL concentration of ribavirin and higher were cytotoxic. Comparison of ivermectin and ribavirin showed that ivermectin was safe at 50[micro]g/mL and lower concentrations. Ribavirin was protective for cell at 12.5[micro]g/mL and 6.25[micro]g/mL only. Comparison of antiviral activity indicated that ivermectin has strong antiviral potential at 100[micro]g/mL and higher but same concentrations were cytotoxic. Ribavirin showed strong antiviral potential at all concentrations

Age and gender specific stroke risk factors in a teaching hospital in Khyber Pakhtunkhwa

To determine age and gender specific stroke risk factors in a teaching hospital in Khyber Pakhtunkhwa. A hospital based descriptive study was performed in medical units of Lady Reading Hospital, Peshawar, from July 2011 to March 2012. A total of 100 patients of 18 years and above and of both genders were included. Study patients with first-ever acute stroke were divided into three groups; Group A young adult [<40 years], Group B, middle-aged [40-60 years], and Group C elderly [>60 years]. In these 03 groups, stroke risk factors were compared. Patients were assessed for hypertension, ischemic heart disease, hyperlipidemia, atrial fibrillation, diabetes mellitus and smoking. SPSS version 18.0 was used for statistical analysis. A sample of 100 patients was taken comprising 68% males and 32% females. Study patients had a mean age of 63.44 +/- 13.849. There were 08 patients in Group A, 29 in Group B, and 63 patients in Group C. Overall, hy-pertension was found in [62%], followed by smoking [47%], coronary artery disease [33%], diabetes mellitus [28%], hyperlipidemia [24%] and atrial fibrillation [21%]. Pattern of distribution of stroke risk factors is age and gender specific. Majority were found to have hypertension. Hypertension, diabetes mellitus, atrial fibrillation and hyperlipidemia differ significantly among the 03 age groups. Smoking was significantly different in males and females

Comparative assessment of extraction methods and quantitative estimation of luteolin in the leaves of Vitex negundo Linn. by HPLC

OBJECTIVE@#To find out the ideal organic solvent and extraction technique for the isolation of luteolin from the leaves of Vitex negundo Linn. (V. negundo) by quantitative estimation of luteolin through high performance liquid chromatography (HPLC) method.@*METHODS@#The leaves of V. negundo were identified by a botanist, cleaned, dried under shade and powdered. Maceration, reflux, Soxhlet and ultrasound assisted extraction techniques were used for the extraction of luteolin from the leaves by using four different solvents of varying polarity such as methanol, ethanol, chloroform, and dichloromethane. A simple HPLC method was used to determine the quantity of luteolin in each sample extract.@*RESULTS@#The calibration plot of standard luteolin showed a linear relationship in the concentration range of 100-500 μg/mL with a correlation coefficient, r(2) of 0.998. The methanolic extract was found to contain highest amount of luteolin and among various techniques employed for extraction and isolation of luteolin, reflux technique was observed to be the most efficient.@*CONCLUSION@#Based on the HPLC results, it can be concluded that reflux technique using methanol is better than the other extraction techniques and should be preferred for the extraction and isolation of luteolin from V. negundo leaves extract in research labs or industries.