Chitinase of Trichoderma longibrachiatum for control of Aphis gossypii in cotton plants.

Chitinase-producing fungi have now engrossed attention as one of the potential agents for the control of insect pests. Entomopathogenic fungi are used in different regions of the world to control economically important insects. However, the role of fungal chitinases are not well studied in their infection mechanism to insects. In this study, Chitinase of entomopathogenic fungi Trichoderma longibrachiatum was evaluated to control Aphis gossypii. For this purpose, fungal chitinase (Chit1) gene from the genomic DNA of T. longibrachiatum were isolated, amplified and characterised. Genomic analysis of the amplified Chit1 showed that this gene has homology to family 18 of glycosyl hydrolyses. Further, Chit1 was expressed in the cotton plant for transient expression through the Geminivirus-mediated gene silencing vector derived from Cotton Leaf Crumple Virus (CLCrV). Transformed cotton plants showed greater chitinase activity than control, and they were resistant against nymphs and adults of A. gossypii. About 38.75% and 21.67% mortality of both nymphs and adults, respectively, were observed by using Chit1 of T. longibrachiatum. It is concluded that T. longibrachiatum showed promising results in controlling aphids by producing fungal chitinase in cotton plants and could be used as an effective method in the future.

Molecular Monitoring of BCR-ABL Fusion Transcripts in Patients with Chronic Myeloid Leukemia During Treatment Using the Endpoint Fluorescence Method.

OBJECTIVE: The purpose of the study was to compare results and evaluate the agreement between the endpoint fluorescence (EPF) method and quantitative real-time polymerase chain reaction (QPCR) during molecular monitoring of patients with chronic myeloid leukemia (CML) receiving treatment. MATERIALS AND METHODS: The study was conducted at Molecular Lab of Riphah International University, Islamabad, Pakistan, from January 2017 to December 2018. A total of 150 blood specimens from 30 patients with CML were analyzed at regular intervals during therapy. The detection/quantification of transcript mRNA was done simultaneously using QPCR and the EPF method. RESULTS: Out of a total of 150 RNA specimens analyzed, 117 (78%) specimens were positive, whereas 33 (22%) were negative for the transcript using both methods at various stages of treatment. Strong linear negative correlations between the cycle threshold and relative fluorescence unit values were observed with P <.0001 at 0, 3, 6, 9, and 12 months of treatment. No significant difference (P >.05) between the means of the BCR-ABL percentage was observed in either method at all stages of treatment. The bias between the 2 methods was calculated as 0.069 ± 3.50, and 95% limits of agreement were 6.92% to -6.79%. CONCLUSION: We found that EPF is s simple method to detect/quantify BCR-ABL mRNA expression during treatment with comparable results to QPCR.

Characteristics of BCR-ABL gene variants in patients of chronic myeloid leukemia.

BACKGROUND: Depending on breakpoints of rearrangement different types of BCR-ABL fusion protein can be generated in patients of chronic myeloid leukemia (CML). The aim of this study is to observe frequencies of major transcripts in CML patients by reverse transcriptase polymerase chain reaction (RT-PCR) and their hematological features at the time of presentation. MATERIALS AND METHODS: This cross sectional study was performed at Molecular Lab of Riphah International University, Islamabad from January to June 2019. Consecutive peripheral blood samples of 70 newly diagnosed CML patients in chronic phase were analyzed by RT-PCR to detect different BCR-ABL transcripts. Routine blood cell counts were assessed by an automated hematology analyzer. RESULTS: All samples expressed typical BCR-ABL rearrangement. Expression of either e14a2 or e13a2 transcript was detected in 38 (54%) and 30 (43%) patients, respectively. Coexpression of e13a2 + e14a2 was found in 2 (3%) patients. The mean total leukocyte count was higher in group expressing e13a2 (P = 0.01). Higher mean platelet count was noted in patients with e14a2 transcript, but this difference was statistically insignificant (P = 0.1). The association of male gender was observed with the group exhibiting e14a2 (P = 0.01). There was no statistically significant association between transcript type and different ranges of age, hemoglobin levels, and platelet and total leukocyte counts (P > 0.05). CONCLUSION: e14a2 transcript was most common transcript in CML patients. Patients exhibiting e13a2 subgroup presented with significantly higher mean white blood cell count at the time of presentation. Significantly higher proportion of male patients was found to express e14a2 transcript over e13a2.

Phenotypic characterization of exotic tomato germplasm: An excellent breeding resource.

Tomato production in Pakistan faces significant problems of low yields due to various biotic and abiotic stresses primarily because of a narrow genetic base of the cultivars being used. Therefore, Introduction and evaluation of the exotic tomato germplasm has become necessary to acquire elite material to develop future breeding programs. To this end, the present study was conducted for the phenotypic characterization of twenty exotic tomato genotypes along with two locally grown cultivars in semi-arid subtropical climate. Data were collected for morphological, fruit quality and fruit yield traits. A significant (p<0.05) phenotypic variation was observed for all the studied traits. Maximum yield was obtained from "Rober" i.e., 1508.31 g per plant. The maximum shelf life was observed in the Cromco, with the least weight loss (2.45%) and loss in the firmness of fruit (22.61%) in 4 days. Correlation analyses revealed a strong genetic association among morphological and yield related traits. High estimates of the heritability (ranged from 79.77% to 95.01% for different traits), along with a high genetic advance (up to 34%) showed the potential usefulness of these traits and genotypes to develop breeding programs to improve the tomato yield and fruit quality.

Serum hepcidin levels in patients with end-stage renal disease on hemodialysis.

Patients on hemodialysis (HD) are usually anemic because of defective erythropoeisis. Hepcidin is a polypeptide that regulates iron homeostasis and could serve as an indicator of functional iron deficiency in patients with end-stage renal disease (ESRD); this may also aid in the assessment of patient's response to erythropoietin (EPO). The present study was directed to investigate serum levels of hepcidin, iron status and inflammation markers such as C-reactive protein (CRP) in patients with ESRD on maintenance HD and to observe the correlation of serum hepcidin with conventional iron and inflammatory markers. A total of 42 patients of both sexes on maintenance HD and EPO therapy were enrolled; 42 ageand sex-matched healthy subjects were included as controls. Laboratory tests including complete blood count, serum hepcidin, total iron binding capacity (TIBC), serum ferritin, serum iron and CRP were performed. Serum hepcidin levels were significantly higher in patients with ESRD than in the control group (18.2 ± 2.8 ng/mL and 8.5 ± 2.3 ng/mL, respectively P = 0.000). The hemoglobin, hematocrit, serum iron, TIBC and transferrin saturation levels in the patient group were significantly lower than in the control group. Higher hepcidin levels were found in EPO non-responders (19.6 ± 2.4 ng/mL) while lower levels (16.9 ± 2.5 ng/mL) were seen in responders (P = 0.001). A positive and significant correlation was observed between the values of serum hepcidin and CRP. Our study indicates that higher hepcidin levels are found in ESRD patients on HD and in those not responding to EPO. Our findings suggest that hepcidin might play a role in the pathophysiology of anemia associated with chronic diseases as well as EPO resistance.

Coagulation factors and antithrombin levels in young and elderly subjects in Pakistani population.

Aging is associated with increased levels of coagulation factors and decrease in natural anticoagulant factors. This strongly supports that age-related hypercoagulable state occurs in elderly. This study aimed to measure the plasma levels of coagulation factors and anticoagulant levels in young and elderly to observe the effect of age on haemostatic system. Ninety healthy individuals, both men and women were divided into two groups on the basis of age. Group I included participants of less than 40 years of age, whereas, group II comprised of participants more than 60 years of age. Fibrinogen activity was assessed by using Clauss technique. Coagulation factor VII, and factor VIII activity by corresponding one stage assay based on prothrombin time and activated partial thromboplastin time. Antithrombin III was measured by the chromogenic method. Our results showed that significantly increased levels of fibrinogen (P = 0.001) were observed in the elderly population as compared to young. Higher fibrinogen levels in younger women than men and comparatively higher level than other races was observed in our local population. Increase in factor VII levels (P = 0.05) was also observed in the elderly group. This increase was statistically significant with age in women (P = 0.03). Factor VIII rose with age in both sexes (P = 0.001). Higher antithrombin activity was observed in the younger group whereas the older group demonstrated significantly lower antithrombin activity (P = 0.001). We conclude that considerable effects of age and sex are observed on coagulation factors and naturally occurring inhibitors.

Frequency of and factors leading to obesity and overweight in school children.

BACKGROUND: Obesity is considered as a global epidemic. Obesity in childhood and adolescent is an issue of concern because it is an important predictor of adult obesity. Identifying individuals who are physically inactive, overweight or obese in school going age is very important. The objective of this study was to determine the frequency and factors leading to obesity in school children. METHODS: A cross-sectional survey was conducted on children aged 13-16 years. It was a questionnaire based survey with measurement of height and weight. RESULTS: Total 431 children were included in the study. Three hundred and nine (71.7%) of the students were in the normal category, i.e., less than 85th centile while 122 (28.3%) had weight greater than 85th centile and were overweight and obese. Positive association was found between eating out and weight categories, eating breakfast, intake of fresh fruits and vegetables and soft drinks, low physical activity, and increased screen time. Frequency of overweight and obese children was high in children from higher socioeconomic status. CONCLUSION: There is strong association between strong determinants of obesity and overweight. Health education of students for known risk factors should be integrated with curriculum of science from elementary classes.