Ghosia Lutfullah

Fungi as chemical industries and genetic engineering for the production of biologically active secondary metabolites

Fungi is somewhere in between the micro and macro organisms which is a good source of producing biologically active secondary metabolites. Fungi have been used as tool for producing different types of secondary metabolites by providing different nutrients at different laboratory conditions. The fungi have been engineered for the desired secondary metabolites by using different laboratory techniques, for example, homologous and heterologous expressions. This review reported how the fungi are used as chemical industry for the production of secondary metabolites and how they are engineered in laboratory for the production of desirable metabolites; also the biosynthetic pathways of the bio-organic-molecules were reported.

Comparative Study of Heavy Metals in Dried and Fluid Milk in Peshawar by Atomic Absorption Spectrophotometry

Various essential and toxic heavy metals (Ca, Mg, Cu, Zn, Fe, Mn, Pb, Cd, Cr, and Ni) contents in various types of dried (infant formula and powdered) and fluid (fresh and processed) cow milk were assessed by atomic absorption spectrophotometry. The milk samples were collected from local markets of different parts of Peshawar city, Pakistan. Heavy metal concentrations varied significantly depending upon the type of milk. The heavy metal concentrations in most of the samples were within normal and permissible ranges. It was observed that the samples contained considerable amounts of calcium, while magnesium levels were well above the required levels. The results also revealed that copper levels were slightly lower than the permissible limits. The concentration of zinc in dried milk samples was greater than the values for the liquid milk types. Infant milk formulae had higher iron levels as compared to other milk samples because of the added constituents. Significant differences were observed in the mean values of manganese and cadmium in different types of milk. The toxic metals were within the acceptable limits and did not show significant levels leading to toxicity.

Structural Bioinformatics of Vibrio cholerae Aminopeptidase A (PepA) Monomer

Aminopeptidase A (PepA) is a metalloexopeptidase found in Vibrio cholerae .It functions as a transcriptional repressor in regulatory cascade that controls virulence gene expression in V. cholerae. It is involved in protein degradation and in the metabolism of biologically active peptides. We proposed a 3D model of PepA based upon the crystal structure of PepA from Escherichia coli (E. coli) with an intention to evaluate the active site of the enzyme and to predict the properties of this enzyme, study of its 3D structure will help in understanding its role in DNA binding.

Study of the genetic traits associated with antibiotic resistance in Staphylococcus aureus isolated from skin wards of Khyber Pakhtunkhwa, Pakistan

Abstract Objective To investigate the prevalence of Staphylococcus aureus ( S. aureus ) isolated from skin wards of the hospitals of Khyber Pakhtunkhwa, its resistance against various commonly and commercially available antibiotics, as well as different genetic traits of resistance and their correlations with the phenotypic visible resistance. Methods In the present study a simple PCR technique were used to investigate the genetic traits of resistance in S. aureus isolated from skin wards of two major hospitals of Khyber Pakhtunkhwa, Pakistan. A total of 100 samples were collected from both the male and female, of which 50 were from patients site of infection and 50 from ward environment. Results These results demonstrated that the total prevalence of S. aureus both in ward as well as in patients was 48%. The S. aureus prevalence was the highest in female patients (50%) followed by ward environment (29%) and then male patients (21%). The antibiotic sensitivity tests revealed that the highest (91.6% isolates) sensitivity was shown to imipenem. However, the highest resistance was found to be against penicillin (100% isolates) followed by cefotaxime (75% isolates). In addition, only 29% of the isolates were found to be resistant to methicillin. PCR technique based on the previously designed primers targeting different genetic traits of resistance revealed that 13 out of the 14 isolates resistant to methicillin were positive for mecA gene. blaZ Genetic traits were found in all isolates resistant to penicillin. The multi-drug resistance traits, vgaA and vgaB each was detected only in 12.5% of S. aureus isolates. The phenotypic character of antibiotic resistance is highly correlated to different genetic traits of resistance. Conclusions Based on our findings, it is concluded that antibiotic resistance in S. aureus strains is increasing day by day due to self-medications and medication by non-registered medical practitioners. Therefore, for quick and fast detection, we propose next-generation sequencing be utilized to screen for antibiotic resistance.

Analysis of oxygen affinity in aquatic amphibian; homology modelling of the major Haemoglobin component HbA1 from the African clawed frog Xenopus laevis, Anura

The homology model of major haemoglobin component HbA1 of the African Clawed Frog was predicted using the pigeon (Columba livia) haemoglobin as a template. The model was built with the help of MODELLER9v8. The models were evaluated with ProSA and PROCHECK. In X. laevis Gln38α is unable to form a hydrogen bond with β97His or β99Asp, which is responsible for the increase in oxygen affinity of the Xenopus HbA1. The hydrogen bond between α34Thr and β124Pro, which stabilises the deoxy state of the haemoglobin, was absent in X. laevis. Hence it is predicted that the HbA1 component of X. laevis has higher oxygen affinity.

Structural Modeling Studies of Aldehyde Dehydrogenase X: Insights into Key Interactions in the Tetrameric Assembly of the Isoenzyme

Human mitochondrial aldehyde dehydrogenase is a member of superfamily of multisubunit enzymes, catalyzing the conversion of a broad range of aldehydes to corresponding acids via the NAD (P) (+)-dependent irreversible reaction. They play an important role in the detoxification of acetaldehyde, in the development of alcohol sensitivity and human alcohol-related disorders. The study aimed to understand the role of conserved residues by comparing similarities and differences between the two isoenzymes. A 3D model of the human ALDHX is constructed by molecular modeling based on the crystal structure of human ALDH2 by using MODELLER (8V1) program. Assessment of reliability of the 3D model is carried out by the programs PROCHECK and PROSAII. The ALDHX fold is similar to the previously described ALDH structures. Sequence and structural analyses have highlighted a close structural and functional relationship between the two isoenzymes of human origin. The interfacial residues that are involved in crucial interactions across the interface stabilize the dimer-tetramer interface in the enzyme. Stability factors like salt bonds and hydrogen bonds aid and maintain the tetrameric assembly of the enzyme.

Herbicidal activity of pure compound isolated from rhizosphere inhabiting Aspergillus flavus

Abstract In the quest for bioactive natural products of fungal origin, Aspergillus flavus was isolated from rhizosphere of Mentha piperita using Potato Dextrose Agar (PDA) and Czapec Yeast Broth (CYB) nutrient media for metabolites production. In total, three different metabolites were purified using HPLC/LCMS and the structures were established using 500 Varian NMR experiments. Further the isolated metabolites in different concentrations (10, 100, 1000 μg/mL) were tested for herbicidal activity using Completely Randomized design (CRD) against the seeds of Silybum marianum and Avena fatua which are major threats to wheat crop in Pakistan. Among the isolated metabolites, one compound was found active against the test weed species whose activity is reported in the present work. The chemical name of the compound is 2-(1, 4-dihydroxybutan-2-yl)-1, 3-dihydroxy-6, 8-dimethoxyanthracene-9, 10(4aH, 9aH)-dione with mass of 388. Results showed that all seeds germinated in control treatment; however, with the metabolite treated, the growth was retarded to different levels in all parts of the weeds. At a dose of 1000 μg/mL of the pure compound, 100% seeds of S. marianum and 60% seeds of A. fatua were inhibited. Interestingly, the pure compound exhibited less inhibition of 10% towards the seeds of common wheat (Triticum aestivum).

Daphne mucronata-mediated phytosynthesis of silver nanoparticles and their novel biological applications, compatibility and toxicity studies

ABSTRACT This contribution reports the biosynthesis of silver nanoparticles (AgNPs) using aqueous leaf extracts of D. mucronata and their diverse applications. Synthesized AgNPs were characterized using diverse techniques, i.e. UV, XRD, EDS, SEM, TEM, FTIR and TGA/DTA. These techniques confirmed the authenticity of the synthesized nanoparticles. The bimodulated AgNPs revealed the highest radical scavenging potential, i.e. 86.4% relative to plant extract at 600 μg/ml. Escherichia coli was found to be the most susceptible strain to AgNPs. Growth of vancomycin-resistant Staphylococcus aureus was also inhibited. Hemolytic activity revealed negligible hemolysis, indicating the biocompatible nature of biomodulated AgNPs. Furthermore, no mutagenic properties were shown by the biogenic AgNPs. Synthesized nanoparticles possessed promising insecticidal potential and had no phytotoxic activity. No haemagglutination was observed for biogenic AgNPs. GRAPHICAL ABSTRACT

Screening of adult dental patients visiting Khyber College of Dentistry, Peshawar for HBV and HCV infections and identifying the associated risk factors

Objective: To screen out adult patients for HBV and HCV infections visiting Khyber College of Dentistry Peshawar (KCD) for different dental treatments and to identify the associated risk factors. Method: This cross-sectional study was conducted at Khyber College of Dentistry, Peshawar in the year 2013. A total of 1540 patients >15 years, visiting KCD for seeking different dental treatments were screened for hepatitis B & C. Informed consent was taken before blood collection and filling of a structured questionnaire. Blood samples were tested against HBsAg and anti HCV by using ICT that were further confirmed by ELISA. The data was analyzed using Epi info version 6. Results: A total of 1540 patients were screened during the study. Among these 36.4%(561) were males and 63.6%(979) were females. Overall prevalence of HBV and HCV was 5.12%(79). On screening, 2.14%(33) were found to be HBs Ag positive of which 2.5%(14) were males and 1.9%(19) were females. HCV was found positive in 2.98%(46) individuals having male to female ratio of 1.6% and 3.8%. Frequency of HBsAg was high in age group 56-65 year and HCV in 36-45 year group. Previous history of IV/IM injections, spouse patient of hepatitis, blood transfusion, surgical operation were found significant risk factors in the transmission of both hepatitis B and C, while previous history of dental treatment and sharing of clippers were significant risk factor in spreading hepatitis C infection only. Conclusions: Overall prevalence of HCV was higher than HBV. Previous history of injections, spouse patient of hepatitis, blood transfusion, surgical operation were found significant risk factors in the transmission of both hepatitis B and C.

Evaluation of Non-Structural Protein-1(NS1) positive patients of 2013 dengue outbreak in Khyber Pakhtunkhwa, Pakistan

Background & Objective: Dengue infection is an arthropod borne disease caused by Dengue virus in humans. Dengue virus infection has more potential to produce severe form of the disease with more severe symptoms. Proper diagnosis of dengue fever is very important for its safe management. The objective of this study was to evaluate the non structural protein-1 (NS1) positive parameter for identification of dengue fever by using ELISA from 2013 dengue outbreak in Khyber Pakhtunkhwa. Methods: It was a cross sectional study conducted among 384 patients tested for dengue admitted to different hospitals of Khyber Pakhtunkhwa April to December 2013 with symptoms related to classical dengue fever. Written informed consent was taken from 100 NS1 positive diagnosed patients, and 3 to 5 ml blood sample was collected for confirmation through ELISA testing. ELISA test for dengue IgG and IgM was performed two time in order to confirm the dengue cases. Data was entered and analyzed by using SPSS version 16. Result: The study performed on 100 NS1 positive samples of patients, admitted to hospitals with symptoms related to classical dengue fever, indicated that after performing the IgM and IgG capture ELISA test only 76 samples were actually found positive for dengue. The rest of the 24 samples were found negative for both IgM and IgG capture ELISAs. The study also revealed that 90.8 % patients had primary dengue infection and 35.5% patients had secondary dengue infection. Most patients were between the age of 10-20 years (26%), among them19.7% were having primary dengue infection. Among 10-20 years of age 50% female patients were false dengue patients. Conclusion: About 24 % NSI protein positive samples were found negative for both IgM and IgG capture ELISAs showed that NS1protein positivity does not confirm actual dengue infection.