volume 9, Issue 1

Methods: In the current study, we determined the role of TP53 mutation in breast cancer in the Punjabi ethnic group/patients of Pakistan. This study was undertaken to determine mutation in the hotspot region of TP53 in breast cancer patients. A total of 30 modified radical mastectomies (MRM) tissue biopsies were collected from different hospitals located in Lahore city of Pakistan. Histopathological analysis was performed to determine the cancer type, stage and grade followed by DNA extraction and sequencing to evaluate mutation in the hotspot region of TP53 gene. Sequencing data was visualized with Bio-edit 7.2.5 and chromas.

Results: According to the Nottingham scoring system 70% patients have grade 3 tumors, followed by grade 2 (23%), grade 4 (6%) and grade-1 (3%). Sanger sequencing analysis of exon 5-9 of TP53 show no mutation. However, many noise mutations were there, which are discussed in detail. This study also shed light that how a genetic analyzer’s results can be correctly analyzed and to avoid false reporting of any mutation which could be a disaster for mis diagnosis of disease. Besides this, a new recipe with very low volume for sequencing PCR ingredients was introduced which could reduce the cost of sequencing.

Conclusion:  The present study suggests that there might be no correlation between TP53 exon 5-9 mutation and breast cancer in the Punjabi ethnic group of Pakistan. However, more studies are required to investigate mutation in the other exons of TP53 gene in large patients’ cohort.  

Method: Dahi was collected from various areas in Lahore. Bacterial cultures were isolated from this local curd and identified using a molecular technique that used 16S rRNA gene primers in PCR and sequencing. The antibacterial assay was carried out using the agar well diffusion method. Out of all the characterized bacteria isolate P26 displayed the best results for the control of pathogenic bacteria. Bacteriocin from isolate P26 was partially purified and its antibacterial assay was performed against sixteen indicator organisms and kanamycin was used as a standard for this experiment. Characterization of bacteriocin was performed at various temperatures, pH, and treating with enzymes.

Result: After the molecular characterization through 16S rRNA, Strain P26 was characterized as Bacillus vallismortis. A protease-sensitive antimicrobial substance was produced by Bacterial strain P26 (Bacillus vallismortis), isolated from local curd (Dahi). This isolate has a broad-spectrum inhibitory activity that can inhibit the growth of both food spoilage and pathogenic bacteria. According to results It was clear that protease-sensitive antimicrobial substance produced from strain P26 is more effective as compared to the antibiotic kanamycin. Maximum activity was shown against Staphylococcus species, a pathogen that is a common cause of gastroenteritis worldwide. This substance was active at 100 ºC for 30 minutes and pH (2-9). The activity was lost after treatment with proteases, so it is classified as bacteriocin.

Conclusion: Bacteriocin P26 is a more effective antibacterial agent than kanamycin, and it can be produced cheaply using local cardkey.  

Methods: A cross-sectional study was carried out for a period of 6 months and a total of 65 patients from the surgical intensive care unit and medical intensive care unit of tertiary care hospital were sampled for the possible pathogen. Non-probability convenient sampling techniques were used to enrolled patients and samples were transported to the pathology department for culture and anti-microbial resistant pattern. Data were analyzed on SPPS version 19 using descriptive and inferential statistical tools. Chi-square test were used for possible association and p value of < 0.05 were considered statistically significant.

Results: Both gram-positive and gram-negative isolates additional to fungi were prevalent in the intensive care unit with the most frequent isolates being Acinetobacter species followed by Enterobacteriaceae and  P. Aeruginosa respectively. Amikacin, Doxycycline, Linezolid, Tigecycline, and Vancomycin have a low resistant pattern for both Staph aureus and coagulase-negative staphylococci (CoNS) while Cephalosporin including cefepime (58%), and minocycline (35%) along with polymyxin B shows the lower resistance pattern for Acinetobacter and other gram-negative isolates. 

Conclusion: Tigecycline, vancomycin, cefepime, and polymyxin B were used as empiric therapy agents in severe conditions however a larger scale study is required for calculating optimized therapeutic regime for each agent. The physician and hospital guidelines should adhere to the monitoring of antimicrobials to limit the upsurge of resistance patterns among different pathogenic organisms.  

Methods: Cassia angustifolia is a well-known herb for pharmaceutical industries as its different compounds are already used in different medicines. Here we performed molecular docking of main compounds of Cassia angustifolia against the main protease of SARS-nCoV2 and were compared with different drugs that are already being used on commercial bases to obtain the lowest energy complex. Auto-Dock vina and its packages were used for molecular docking of SARS-nCov2.

Results: Molecular docking of Cassia angustifolia compounds represent very promising binding energies complexes, e.g., Sennoside B gives -9.05kcal/mol and Aloe-Emodin give -4 Kcal/mol of energy against the main protease of coronavirus. In contrast, a couple of commercially used antiviral drugs were also evaluated against the selected protein of coronavirus e.g., Hydroxychloroquine and Ribavirin complexes appeared with -5.2 Kcal/mol and -6.3 Kcal/mol of energy respectively.

Conclusion: Many compounds of Cassia angustifolia showed the promising energy complexes even better than the commercially used antiviral drugs e.g., Sennoside B which has the best energies against main protease of coronavirus. Further, in-vivo and in-vitro studies are needed to validate this hypothesis with advanced MD simulations and wet-lab experimentations. 

Methods: In total, 120 day-old Hubbard broiler chickens assigned at random to four nutritional treatments for period of 42d trial. The broiler^’s rations (0-28d and 29-42d) were added with 0, 0.050, 0.075 and 0.10% exogenous emulsifier with1.0% soybean oil. The data relating performance, carcass traits, fat digestibility and blood fat profiles in chickens was obtained during experiment.

Results: During days 1-21 and 1-42, chickens fed diets with high levels (0.075 or 0.100%) of emulsifier with soybean oil resulted significantly (p<0.05) better weight gain and feed: gain ratio than without emulsifier. The broilers on emulsifier diets fed substantially (p<0.05) less feed intake and considerable (p<0.05) decreased feed: gain ratio as compared to without emulsifier at 1-21d. The relative weight of the breast or leg meat yield, and liver significantly (p<0.05) increased in broilers given exogenous emulsifier rations. On day 21 and 42, the digestibility of fat in birds offered with emulsifier diets were substantially (p<0.05) increased than that of control group. Lipid profile was not influenced by exogenous emulsifier supplementation in broiler’s diet.

Conclusion: This study envisaged that exogenous emulsifier supplementation (0.05 to 0.10%) in broiler’s diet improved the performance, meat yield and fat digestibility without affecting on cholesterol profile in broilers 

Method: Initially selective bacterial strains were isolated from different waste samples. COD of the three samples of dyes were performed by titration method to observe the pattern of reproducibility of the biodegradability (COD) results.

Results: Textile industry sludge wastewater samples contained different characteristics of dye degrading consortium of bacterial strains as compared to soil extracts samples whereas did not get any bacterial growth in domestic wastewater samples.  It was interesting to observe that the reproducibility of COD results was very hard to get due to sensitivity of the test parameter to the prevailing environmental conditions.

Conclusion: Present study is significant for understanding the sensitivity of reproducibility or validation of biodegradation study. On the basis of present findings of experiments, we can say that only the method of determining COD could be standardized which will ensure the credibility of the results and the changed environmental temperature could affect the controlled experimental results. Present finding would give benefit to the new scientists and students for understanding the normal behavior of biodegradation study. 

Methods: A total of 100 blood samples (1-2 ml) were collected from the hospitals of Lahore. After serum collection, indirect ELISA was performed on a commercially available kit. The Optical density (OD) was taken from the ELISA reader. Positive and negative samples were also run along with the samples and samples OD was compared with the OD of positive and negative controls. Cut off value was calculated by multiplying negative control (NC) with 2.1. The absorbance value of specimen/cutoff <1: samples having a value less than cut-off value were considered negative and samples having a value greater than or equal to cut-off value were considered positive. The apparent prevalence of Hepatitis A virus was measured by dividing the number of children positive to the total number of children included in the study.

Results: Of the 100 samples tested, 37% were positive for IgM and 100% were positive for IgG. Statistical Analysis SPSS 21.0 version was applied to analyze the data and a correlation test was applied to see the association of risk factors with disease status.

Conclusion: This study was done to appraise the overall status of HAV prevalence in young kids in association with potential risk factors. 

Methods: The current study is the first ever reported case of transforming Arabidopsis thaliana with plant expression binary vector pSB219 harboring PTF1 via floral dip method and analyzing phosphorus stress induced genes interaction, through yeast-one-hybrid. Yeast-one-hybrid analysis was performed on four selected genes namely LPR1 (Low Phosphate Response), PDR2 (Phosphate Deficiency Response 2), PHT1;2 (Phosphate Transporter) and RNS1 (Ribonuclease). The positive transformed lines were expression analyzed for PTF1 by real time PCR and further studied for their root morphology.

Results: The results clearly showed direct interaction of LPR1 with PTF1 while other genes, although being overexpressed, were indirectly regulated. Transformation efficiency of 1% was achieved and a maximum 2.5-fold increase in PTF1 expression was observed. Root morphological studies exhibited significantly enhanced root hair and lateral surface area when grown in phosphorus deficient MS medium.

Conclusion: The results of the current study may pave path for improved comprehension of gene interactions and root architecture modifications under phosphorus limiting conditions. 

Methods: Peripheral blood and clinical data were collected from CML patients with durable treatment response, along with appropriate controls. DNA was extracted and whole exome sequencing (WES) carried out to screen novel genes mutated only in experimental groups and absent in control groups. Mutations were confirmed using Sanger sequencing.  Data was analyzed using SPSS version 23.

Results: Although WES detected 10 genes mutated exclusively in CML patients with durable treatment response, Sanger sequencing could confirm mutations only in RAI1 gene (GC deletion at nucleotides 837-838, a frameshift mutation).

Conclusions: Our study shows that mutations in a novel gene (RAI1) are associated with durable response in CML patients.  RAI1 gene is active throughout the body and controls functions of many genes involved in daily rhythms. Our studies provide first important insights into molecular factors associated with long-term treatment response in CML that can serve as novel biomarker to identify patients eligible for TKI cessation in many ongoing CML STOP-TKI trials. 

Methods: A cross sectional study was designed to check the prevalence of thyroid disease and its correlation with vitamin D levels in females. Venous blood was drawn from the female’s patients (11—80 years of age) using gel disposable vials (3.5 ml) in aseptic condition. Samples were centrifuged at four thousand revolutions per minute for five minutes and serums were separated. After the separation of serum, the samples were transferred to the laboratory for the automated estimation of Thyroid Stimulating Hormone (TSH), Triiodothyronine (T3), Thyroxine and Vitamin D.

Results: It was observed that out of 79 females who had gone through thyroid profile testing, 70% females had normal thyroid profile. However, hypothyroidism was found in 23% females and 7% females had the condition of hyperthyroidism. Out of 18 (23%) reported cases of hypothyroidism, 8 cases were of mild (subclinical) hypothyroidism, 3 cases indicated non-thyroidal illness; rare pituitary hypothyroidism. Out of 79 female participants, only 20 (25.31%) females had normal serum vitamin D levels. Overall, 59 (74.68%) females had vitamin D deficiency. When the vitamin D deficiency was correlated with thyroid dysfunction, it was observed that vitamin D levels were non-significantly (p = 0.35) associated with hypothyroidism.

Conclusion: Hypothyroidism was found prevalent in the tested female population, as 23% of the tested population had hypothyroidism while 9% of them have hyperthyroidism. Moreover, majority of the population had vitamin D deficiency. 

Methods: The eggs of Ascaris lumbricoides were isolated from soil samples from an indigenous village in Malaysia, the eggs were isolated using the floatation technique and cultured in vitro using 0.1% of sulphuric acid dish and embryonation of the parasite eggs was observed daily under a light microscope till day 28. For the in vivo method, the embryonated eggs around 400 epg and 800 epg were feed to the rats for four weeks. Finally, stool samples were collected for Ascaris egg shedding.

Results: The loamy eggs collected near toiled areas using the floatation technique were found to contain Ascaris lumbricoides eggs. The cultured eggs in sulphuric acid developed from the first-cell stage until the fourth stage of development. Discharge of Ascaris eggs were found in stool samples among rats fed with 200epg of Ascaris eggs.

Conclusion: It was found that 0.1% sulphuric acid was ideal for developing Ascaris lumbricoides eggs for cultivating in vitro condition, and there was the possibility of rats carrying the Ascaris eggs throughout the village. In future, these findings can be used to propagate the eggs for testing anthelminthic drugs inhibiting the developmental stages. 

Methods: A total of 80 pairs of fingerprints (1600 prints) were collected from persons aged 15 to 30 years using rolling method. Out of which 20 pairs were brother-brother, 20 were sister-sister, 20 were brother-sister and 20 Pairs were random. Each fingerprint was analyzed for the gender identification on the basis of minutiae, ridge density and types. All the fingerprints were analysed using ACE-V method. After comparison SPSS software was used for further analysis.

Results: Our result showed that the types of fingerprints identified was whorl (50%) followed by loop (45.25%), arch (4.5%) and 0.25% of the accidental type. The dominant type was whorl while accidental was the least common type of fingerprints. Statistical analysis showed that between the groups, brother-brother and sister-sister was significant while rest of the groups was not significant. Moreover, greater ridge density was observed in female as compared to male.

Conclusion: It is concluded that the sibling fingerprints had greater similarity as compared to non-sibling, however both male and female fingerprints were significantly different in term of ridges density. This study may be useful in crime scene investigation.  

Methods: A study was conducted in complete randomized design (CRD) using two onion varieties, Phulkaara and Desi Red, under Faisalabad, Punjab conditions. The level of salinity was kept 50 mM NaCl and 100 mM NaCl while 400 ppm of salicylic acid was applied weekly. The data was analyzed through Statix 8.1 to compute analysis of variance (ANOVA).

Results: All the growth and physiological parameter of onion seedlings were decreased significantly with the increasing level of salinity while increased with the application of salicylic acid except total soluble protein. Salicylic acid (@400ppm) mitigated the adverse effects of salinity by enhancing photosynthetic activities. The total soluble protein was increased in leaves with increasing salinity stress whereas the application of SA has non-significant effects. The performance of both varieties remained significantly same for the parameters dry shoot weight, plant height, chlorophyll a, b and total chlorophyll contents. The variety Desi Red performed better for shoot length while Phulkaara was better in fresh shoot/root weight, length of root, dry root weight and total soluble protein.

Conclusion: The application of foliar spray of salicylic acid is potential growth regulator to increase the growth parameters of onion under saline stress conditions.  

Methods: We used genomic and transcriptomic datasets from Pan-Cancer TCGA (The Cancer Genome Atlas), Genotype-Tissue Expression (GTEx) and microarray platforms and conducted in silico analysis of all the genes encoding nucleoporins that are associated with the Nuclear Pore Complexes (NPCs).  For mutation detection, we used cBioportal; for expression analysis, we used Xena and for patient survival plots, KMPlot was utilized.

Results: The genetic and molecular profile of nucleoporin genes identified multiple mutations and detected aberrant expression in breast cancer. Interestingly, NUP133, AHCTF1, TPR, Nup121L showed simultaneous gene amplification in nearly 10% of breast cancer patients. In addition, deregulated expression of some of the nucleoporins, namely, NUP62, NUP 93, NUP98, NUP155, POM121L12, RAE1, SEC13, TPR were correlated with patient prognosis.

Conclusion: The current study is the first one that unravels a comprehensive molecular and genetic profile of nucleoporins genes in breast cancer and underscores the critical roles of various nucleoporins in cancer progression. The identified molecules may advance our understanding of the etiology of the disease and serve as possible targets for novel therapeutic strategies in cancer. 

Methods: mtDNA was extracted from the patients' whole blood. After PCR amplification, the DNA was sequenced and analyzed.

Results: The sequencing results showed the absence of the most common mtDNA mutation m.A3243G in diabetic families with individuals who suffer from hearing loss.

Conclusions: The m.A3243G mutation in the mitochondrial tRNALeu (UUR) Leu gene was not found to be a common cause of type 2 diabetes and deafness. These results suggest that there may be genetic causes for this phenotype.  

Methods: The disc-diffusion and minimal inhibitory concentration (MIC) methods were carried out to determine the antibacterial activity of aqueous and methanol extracts of A. longa roots against several reference bacterial strains.

Results: Both aqueous and methanolic extracts had antibacterial activity to varying degrees with the disc-diffusion assay, however the methanolic extract shown significant activity at a higher dosage (200 mg/ml). Escherichia coli (22.0 mm) was the most sensitive bacterium tested, followed by Staphylococcus aureus (16.0 mm) and Bacillus subtilis (12.0 mm). The bacteria with the lowest susceptibility were Klebsiella pneumonia, Pseudomonas aeruginosa, and Micrococcus luteus, which all had a 10.0 mm zone of inhibition. The MIC values corroborated these observations, with E. coli MICs of 6.25±1.5 mg/ml, S. aureus MICs of 25±1.02 mg/ml, M. luteus MICs of 12.5±1.25 mg/ml, K. pneumonia MICs of 50.0±0.75 mg/ml, and B. subtilis MICs of 100±0.6 mg/ml, respectively. The findings of the methanolic extract were statistically significant and equivalent to those of the antibiotics studied.

Conclusion: Roots of A. longa have significant broad-spectrum antibacterial agents, notably in the methanolic extract, which validated their applications in Moroccan folkloric medicine to treat soft-tissue and skin infections.  

Materials: Ten Fresh venous blood samples were collected from healthy males (20-45 years old) by venepuncture and stored at 2–8^oC until used. Each particular sample was subjected to five blood collection options as follows: the blood was freshly aliquoted without any treatments, the blood was collected in EDTA (ethylene diamine tetra acetic acid) tubes, the blood was collected with the addition of phosphate buffered saline (PBS) in a ratio of 1:2, the blood was added to PBS 1:2 and Triton X 100, and the blood was centrifuged and then PBS 1:2 and Triton X 100 were added. The quantity and integrity of the extracted DNA were evaluated by agarose gel electrophoresis. The quantity and purity of the DNA were measured using a NanoDrop Spectrophotometer.

Results: The current study indicated statistically significant difference (p ≤0.05) among the treated groups when the concentration of the extracted DNA was taken into consideration. There was a statistically significant difference among the five groups when their absorbance ratio was measured at 260 nm/ 230 nm (p ≤0.05). However, no statistically significant difference (p ≤0.05) was obtained among the groups when the absorbance ratio of 260 nm/280 nm was considered.

Conclusion: The pretreatment of blood samples with buffers (PBS containing 5% Triton X100) prior to DNA extraction can lead to an altered DNA yield and purity.