Volume 7, issue 3

Methods: In this study, an endochitinase gene was isolated from Trichoderma harzianum, and characterized in-silico by using various bioinformatics tools. Further, the gene was cloned in eukaryotic expression vector (pPICZA) under the control of AOX1 promoter for recombinant expression in Pichia pastoris GS115 host strain.

Results: The chitinase cDNA was ~1000 bp long, while in-silico studies revealed an open reading frame of 888 bp encoding 295 amino acids with a calculated molecular mass of 37332.76 Da and an estimated isoelectric point of 4.07. Recombinant chitinase protein expressed intracellularly and revealed high expression in P. pastoris host. The 37 kDa recombinant chitinase protein developed with antigen antibody confirmed its expression in P. pastoris.

Conclusion: Conclusively, T. harzianum derived chitinase gene was successfully over expressed in P. pastoris where recombinant protein was expressed intracellular in the form of inclusion bodies. 

Methods: Variant calling and associated workflows were executed on NGS pooled-seq data of 12 Pakistani Teddy goats. Different tools used in this pipeline are FastQC for quality checks, Trimmomatic for trimming data, SAM/BAM tools for conversion of file formats, Picard tools for marking deduplicates, VCFtools/FreeBayes for genomic variant detection and SnpSift to annotate the variants.

Results: Highly associated functionally untrivial 43,712 loci were percolated having 87,510 alleles. Besides, 1,548 variants with 1,134 SNPs, 23 mixed variants, 76 MNP, 183 insertions and 132 deletions were observed in Teddy breed using San Clement ARS1 reference genome. Furthermore, 1,283 homozygous and 265 heterozygous variant were also divulged out of 43,447 loci. These variants are likely to be liable for general phenotypic traits of Teddy with smaller body-size, tender meat quality and agility along with other breed specific traits. 

Conclusion: Galaxy fulfills the core function of reproducibility and easy accessibility by removing the gaps between large data analysis and its interpretations. This variant calling pipeline reveals the genomic differences of Teddy specific characteristics as compare to ARS1 reference genome. 

Methods: A total of five fractions were selected i.e. crude methanolic extracts, n-hexane, chloroform, ethyl acetate and aqueous extracts were prepared and their activity checked against four bacterial strains including Streptococcus mutans, Staphylococcus aurous, MRSA (methicillin resistance Staphylococcus aurous) and Serratia marcescens; and four fungal strains i.e. Fusarium oxysporum, Aspergillus flavus, Polysphondylium pallidum and Alternaria alternata.

Results: The highest anti-bacterial activity shown by crude methanolic extract fraction i.e. (48-38%) and the lowest activity was exhibited by aqueous extract (15-0%) against all the selected bacterial strains. Similarly, the highest anti-fungal activity indicated by crude methanolic extracts (60-45%) against the four selected fungal species and lowest activity shown by the aqueous fractions (26-0%). Reasonable activity was also exhibited by others fractions as well.

Conclusion: The outcomes of our experiment strongly supports that Moringa oleifera has significant antibacterial and antifungal activities, so the plant is effective antibacterial and antifungal agent. 

Methods: The total 239 rhizobacterial isolates were screened with agar plug and well diffusion methods, characterized biochemically and identified by Bergey’s manual scheme. In chemical screening, metabolic extracts of rhizobacterial isolates were separated in thin layer chromatography (TLC) and further identified by HPLC. The ability of rhizobacterial strains for optimum metabolite production was checked in vitro under different environmental conditions.

Results: Results showed that 31% isolates had antibacterial activity against Bacillus cereus, Bacillus subtilis, E. coccus, E. coli, Salmonella, Klebsiella, Staphylococcus aureus ATCC20737, Staphylococcus aureus ATCC25923 and Neisseria. Gonorrhea ATCC19424, methicillin resistant Staphylococcus aureus 1, methicillin resistant Staphylococcus aureus 6 and methicillin resistant Staphylococcus aureus 8.  Maximum rhizobacterial strains showed best antibacterial activity at 35^ᵒC and at pH7. ST₅ strain showed significant results against E. coli ETCC and methicillin resistant Staphylococcus aureus-7 (25mm zone of inhibition). The bands of ST₅ and ST₂ extracts on a TLC plate were detected at Rf value 4.7 and 4.8 respectively. In HPLC, different peaks at different retention times with maximum peak at 2.90 and 2.92 indicating the production of antimetabolites.

Conclusion: These rhizobacterial isolates are valuable inputs in natural consortium to produce specific antibacterial metabolites. Demonstrating the structure and purification of these bioactive compounds may enhance the antimicrobial activities which further can be helpful in development of antibiotics. 

Methods: The clinical confirmed Ps. aeruginosa samples were collected according to the standard protocol, at Khyber Teaching Hospital (KTH), Peshawar. All collected samples were sub- cultured on appropriate culture media. After isolation and identification, the antibiotics susceptibility testing was performed. The detection of ESBL was carried out by the double-disc diffusion method. Carbapenemase-producing bacteria was confirmed by the modified Hodge test. Descriptive analysis was performed for statistical analysis of collected data.

Results: A total of one hundred and sixty-two (n=162) Ps. aeruginosa confirmed isolates were collected, in which 59.3% were male and 40.7% were from female patients. The percentages of ESBL and carbapenemase producing Ps. aeruginosa isolates were 5.5% and 23.5%, respectively. The multidrug resistance was observed against 27.2% isolates. Among tested antibiotics highest percentages of resistance was observed against ciprofloxacin (43%) and ceftazidime (39.5%). 

Conclusion: We observed highest level of drug resistance in Ps. aeruginosa clinical isolates against tested antibiotics and majority of the isolates were Multi-drug resistant (MDR). 

Methods: In this view, a research experiment was designed at Sugarcane Research Institute, Faisalabad during 2016 and 2017, to study the feasibility of bud chips as an alternate to conventional planting in sugarcane crop. The experiment was planted with sugarcane variety CPF-252 in randomized complete block design with split plot arrangement having three replications.

Results: The results of study revealed that setts planting on 15 April gave maximum cane (61 t ha^-1) and sugar yield (7.15 t ha^-1). While bud chips sown on 15 April produced higher yield. The benefit cost ratio (BCR) was high in bud chips planting (1.63) than setts planting of sugarcane.

Conclusion: The study concludes that bud chip planting on 15 April is an improved technique with greater potential to provide disease free seed, improve the yield with minimum quantity of seed than conventional planting. 

Method: Eye colour digital photographs and buccal swab samples were collected from 267 Pakhtun individuals of District Swat. Any person with eye disease was excluded from the study. Genomic DNA was extracted through Phenol-Chloroform extraction method. The amplified SNPs were typed using Multiplexed Single Base Extension (SBE). The genotypes were checked for eye color phenotypes through IrisPlex online tool and correlation were checked between SNPs, Gender, pie score and eye color.

Result: Brown eye color was found prevalent as compared to intermediate and blue. Females have highly brown eye color compared to males while males have intermediate and blue. Three SNPs rs12913832 (in the HERC2), rs1393350 (TYR gene), rs1800407 (OCA2 gene) were strongly significant to eye color. Pie score was also significant to eye color and rs12913832 SNP. IrisPlex analysis in 20 individuals of District Swat was performed. The prediction accuracy of IrisPlex for blue or brown was 100% in the studied individuals. However, the IrisPlex tool predicted the intermediate phenotype incorrectly as brown or blue.

Conclusion: It is concluded from the data that intermediate eye colour was not predicted accurately, therefore, inclusion of more SNPs in the IrisPlex system is needed to predict intermediate eye colour accurately. 

Methods: A prospective study was carried out on seventy five, normal BMI subjects aged between 20–50 years. CT imaging was used for volumetric measurement of TAT, SAT and VAT. Pearson’s correlation of these were then found out with age. Kruskal Wallis test was also performed to compare these in hypertensive and non-hypertensive subjects and in those with different physical activity levels (PAL).

Results: Women showed significantly higher volumes of TAT and SAT. Men showed statistically significant correlations of TAT and VAT with age. SAT volumes had significant negative association with the PAL in both genders. Men showed higher responsiveness of fat deposition in all compartments to the presence of hypertension.

Conclusion: In conclusion, factors such as gender, age, level of physical activity and hypertension affect the site specific deposition of fat even in those individuals who aren’t over-weight or obese. 

Methods: Rhizosphere soil samples were collected from rhizosphere of soil plantation areas, to be used for bacteria isolation. The identified bacteria were qualitatively tested for their ability to produce slime and subsequently develop biofilm.

Results: The cultural and biochemical identification techniques, including morphological, biochemical and molecular methods revealed that the antagonistic bacteria- from the distinctive rhizosphere soil samples belong to Pseudomonas genus in particular, P. aeruginosa (PF1a, pf2a, PF-8) and P. putida (PF-7). These identified isolates inhibited Aspergillus niger development with percentage of parasitic growth inhibition greater than (48.095 ± 2.182)% for P. aeruginosa (pf-8). In addition, these identified isolates were significantly shown to be able to produce exopolysaccharide and subsequently develop biofilm on polystyrene and glass surfaces.

Conclusion: Superior strains of these bio-control and plant growth promoting rhizobacteria will enable for better biological control of fungal and bacterial plant diseases and may reduce chemical pesticide usage. The indigenous strains isolated could potentially have a great impact on controlling plant diseases, in particular, those caused by microorganisms, and could be used as an alternative bio control agents instead of harmful chemical pesticides. Most of the tried microbes produced exopolysaccharides as well as formed biofilm on polystyrene and glass surfaces. 

Methods: Current reading describes a study conducted on 50 breast cancer patients from Institute of Nuclear Medicine and Oncology (Lahore). The patients were clinically categorized as stage II (19 patients) and stage III (29 patients) respectively. Most of the patients were diseased with infiltrative ductal carcinoma (48) of the breast. Blood samples of all patients were collected after forty days of chemotherapy course besides 10 healthy subjects.

Results: There was a significant rise in LDH (221.48 ± 18.44) and Ferritin (356.46 ± 12.28) levels as compared to control groups. Concentration of GGT (34.12 ± 1.61) was also elevated but not significantly raised. There was no significant rise in ALP (279.27 ± 26.60), AST (49.44 ± 8.510), ALT (33.40 ± 2.83), bilirubin (4.87 ± 3.96), Hb (11.16 ± .299) and platelets (032.54 ± 21.68) levels in comparison to normal control subjects. An elevation of LDH and ferritin levels in cases of carcinoma breast signifies its importance as a biomarker of disease. A serial measurement of these enzymes would have diagnostic and prognostic significance and help treatment decisions.

Conclusion: The ferritin along with lactate dehydrogenase can be used as a valuable biomarker for breast cancer diagnosis and prognosis.