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Tomato mosaic virus (ToMV) is one of the lethal tomato pathogens. In this study, we collected tomato leaves exhibiting interveinal chlorosis from farmer’s fields in district Lahore, Pakistan. The presence of tobamoviruses was confirmed through extracting RNA, cDNA synthesis, PCR amplification and subsequent sequencing of capsid protein (CP) gene of ToMV that contained 480 nucleotides (encoded for 160 amino acids making ~17 kDa protein) located at 5703rd to 6182nd nucleotide position in ToMV genome. In sequence analysis, Pakistani isolates shared 98.9-99.5% and 97.3-99.3% nucleotide and amino acid homologies with each other and 96-99% and 90-98.6% with other isolates respectively. In phylogenetic analysis the Pakistani isolates grouped with Iranian and Chines isolates, exhibiting higher nucleotides and amino acids identities as well. This research reveals a geographical association among Pakistani, Chines and Iranian ToMV isolates. The study suggests extensive surveys of tobamoviruses infecting field crops for their detailed genetic information.

Heat stress is considered to induce a wide range of physiological and biochemical changes that cause severe damage to plant cell membrane, disrupt protein synthesis, and affect the efficiency of photosynthetic system by reducing the transpiration due to stomata closure. A brief and mild heat shock is known to induce acquired thermo tolerance in plants that is associated with concomitant production of heat shock proteins’ (HSPs) gene family including HSP70. The findings from different studies by use of technologies have thrown light on the importance of HSP70 to heat, other abiotic stresses and environmental challenges in desserts. There is clear evidence that under heat stress, HSP70 gene stabilized the membrane structure, chlorophyll and water breakdown. It was also found that under heat stress, HSP70 decreased the malondialdehyde (MDA) content and increased the production of superoxide dismutase (SOD) and peroxidase (POD) in transgenic plants as compared to non-transgenic plants. Some reactive oxygen species (ROS) such as superoxide, hydrogen peroxide and hydroxyl radical are also synthesized and accumulated when plants are stressed by heat. Hence HSP70 can confidently be used for transforming a number of heat tolerant crop species.

Hepatitis is one of the common liver diseases, imposing a heavy health burden worldwide. Acute hepatitis may develop into chronic hepatitis, progressing to cirrhosis and hepatocellular carcinoma. In the present study, the expression of miRNAs was quantified by real-time PCR, such as miRNA-182, 122, 21, 150, 199, and 222. Along with the control group, HCV was divided into chronic, cirrhosis, and HCC groups. The treated group was also included after the successful treatment of HCV. Biochemical parameters, such as ALT, AST, ALP, bilirubin, viral load, and AFP (HCC), were also evaluated in all of the study groups. We compared the control and diseased groups; these parameters showed significant results (p = 0.000). The viral load was high in HCV but was not detected after treatment. miRNA-182 and miRNA-21 were overexpressed with disease progression, while the expression of miRNA-122 and miRNA-199 was increased compared with the control, but decreased in the cirrhosis stage compared with chronic and HCC. The expression of miRNA-150 was increased in all of the diseased groups compared with the control, but decreased compared with the chronic group. We compared the chronic and treated groups and then all of these miRNAs were down-regulated after treatment. These microRNAs could be used as potential biomarkers for diagnosing different stages of HCV.

The SARS-CoV-2 pandemic continues to expand globally, with case numbers rising in many areas of the world, including the Indian sub-continent. Pakistan has one of the world’s largest populations, of over 200 million people and is experiencing a severe third wave of infections caused by SARS-CoV-2 that began in March 2021. In Pakistan, during the third wave until now only 12 SARS-CoV-2 genomes have been collected and among these nine are from Islamabad. This highlights the need for more genome sequencing to allow surveillance of variants in circulation. In fact, more genomes are available among travellers with a travel history from Pakistan, than from within the country itself. We thus aimed to provide a snapshot assessment of circulating lineages in Lahore and surrounding areas with a combined population of 11.1 million. Within a week of April 2021, 102 samples were sequenced. The samples were randomly collected from two hospitals with a diagnostic PCR cutoff value of less than 25 cycles. Analysis of the lineages shows that the Alpha variant of concern (first identified in the UK) dominates, accounting for 97.9 % (97/99) of cases, with the Beta variant of concern (first identified in South Africa) accounting for 2.0 % (2/99) of cases. No other lineages were observed. In depth analysis of the Alpha lineages indicated multiple separate introductions and subsequent establishment within the region. Eight samples were identical to genomes observed in Europe (seven UK, one Switzerland), indicating recent transmission. Genomes of other samples show evidence that these have evolved, indicating sustained transmission over a period of time either within Pakistan or other countries with low-density genome sequencing. Vaccines remain effective against Alpha, however, the low level of Beta against which some vaccines are less effective demonstrates the requirement for continued prospective genomic surveillance.

SARS-CoV-2 is a causative agent for COVID-19 disease, initially reported from Wuhan, China. The infected patients experienced mild to severe symptoms, resulting in several fatalities due to a weak understanding of its pathogenesis, which is the same even to date. This cross-sectional study has been designed on 452 symptomatic mild-to-moderate and severe/critical patients to understand the epidemiology and clinical characteristics of COVID-19 patients with their comorbidities and response to treatment. The mean age of the studied patients was 58 ± 14.42 years, and the overall male to female ratio was 61.7 to 38.2%, respectively. In total, 27.3% of the patients had a history of exposure, and 11.9% had a travel history, while for 60% of patients, the source of infection was unknown. The most prevalent signs and symptoms in ICU patients were dry cough, myalgia, shortness of breath, gastrointestinal discomfort, and abnormal chest X-ray (p < 0.001), along with a high percentage of hypertension (p = 0.007) and chronic obstructive pulmonary disease (p = 0.029) as leading comorbidities. The complete blood count indicators were significantly disturbed in severe patients, while the coagulation profile and D-dimer values were significantly higher in mild-to-moderate (non-ICU) patients (p < 0.001). The serum creatinine (1.22 μmol L-1; p = 0.016) and lactate dehydrogenase (619 μmol L-1; p < 0.001) indicators were significantly high in non-ICU patients, while raised values of total bilirubin (0.91 μmol L-1; p = 0.054), C-reactive protein (84.68 mg L-1; p = 0.001), and ferritin (996.81 mg L-1; p < 0.001) were found in ICU patients. The drug dexamethasone was the leading prescribed and administrated medicine to COVID-19 patients, followed by remdesivir, meropenem, heparin, and tocilizumab, respectively. A characteristic pattern of ground glass opacities, consolidation, and interlobular septal thickening was prominent in severely infected patients. These findings could be used for future research, control, and prevention of SARS-CoV-2-infected patients.

SARS-CoV-2 is a causative agent for COVI-19 disease, initially reported from Wuhan, China. Infected Patients experienced mild to severe symptoms, resulting in several fatalities due to a weak understanding of its pathogenesis, which is the same even to date. This cross-sectional study has been designed on four hundred and fifty-two symptomatic, mild-to-moderate, and severe/critical patients to understand the epidemiology and clinical characteristics of COVID-19 patients with their comorbidities and response to treatment. The mean age of studied patients was (58±14.42) years, and the overall male to female ratio was 61.7 to 38.2%, respectively. 27.3% of the patients had a history of exposure, 11.9% travel history, while for 60% of patients, the source of infection was unknown. The most prevalent signs and symptoms in ICU patients were dry coughs, myalgias, shortness of breath, gastrointestinal discomfort, and abnormal Chest X-ray (pp=0.007) and COPD (p=0.029) as leading comorbidities. Complete Blood Counts indicators were significantly increased in severe patients, while the Coagulation Profile and D-dimer values were significantly higher in mild-to-moderate (non-ICU) patients (p < 0.001). Serum Creatinine (1.22 umole L-1; p = 0.016) and LDH (619 umol L-1; p < 0.001) indicators were significantly high in non-ICU patients while, raised values of Total Bilirubin (0.91 umol L-1; p = 0.054), CRP (84.68 mg L-1; p = 0.001) and Ferritin (996.81 mg L-1; p < 0.001) were found in ICU patients. Drug Dexamethasone was the leading prescribed and administrated medicine to the COVID-19 patients, followed by Remdesivir, Meropenem, Heparin, and Tocilizumab, respectively. A characteristic pattern of Ground glass opacities (GGO), consolidation, and interlobular septal thickening were prominent in severely infected patients. These findings could be used for future research, control, and prevention of SARS-CoV-2 infected patients.

BackgroundThe SARS-CoV-2 pandemic continues to expand globally, with case numbers rising in many areas of the world, including the Indian sub-continent. Pakistan has one of the world ‘s largest population, of over 200 million people and is experiencing a severe third wave of infections caused by SARS-CoV-2 that begun in March 2021.In Pakistan, during third wave until now only 12 SARS-CoV-2 genomes have been collected and among these 9 are from Islamabad. This highlights the need for more genome sequencing to allow surveillance of variants in circulation. In fact more genomes are available among travellers with a travel history from Pakistan, than from within the country itself.MethodsFor a better understanding of the circulating variants in Lahore and surrounding areas with a combined population of 11.1 million, within a week of April 2021, 102 samples were sequenced. The samples were randomly collected from 2 hospitals with a diagnostic polymerase chain reaction (PCR) cutoff value of less than 25 cycles.ResultsAnalysis of the lineages shows that B.1.1.7 (first identified in the UK, Alpha variant) dominates, accounting for 97.9% (97/99) of cases, with B.1.351 (first identified in South Africa, Beta variant) accounting for 2.0% (2/99) of cases. No other lineages were observed.DiscussionIn depth analysis of the B.1.1.7 lineages indicates multiple separate introductions and subsequent establishment within the region. Eight samples were identical to genomes observed in Europe (7 UK, 1 Switzerland), indicating recent transmission. Genomes of other samples show evidence that these have evolved, indicating sustained transmission over a period of time either within Pakistan or other countries with low density genome sequencing. Vaccines remain effective against B.1.1.7, however the low level of B.1.351 against which some vaccines are less effective demonstrates the requirement for continued prospective genomic surveillance.

Background: Drought stress seriously affects the cotton fiber development. Universal stress protein gene isolated from native species Gossypium arboreum has the promising tolerance role against these stresses. Objectives: This study aimed to clone, characterize, and genetically transform the GUSP1 gene in local cotton and to observe its expression in transgenic plants under drought stress. Materials and Methods: Universal Stress Protein (GUSP1) gene from Gossypium arboreum was cloned in pCEMBIA (-) 1301plant expression vector by replacing Hygromycin and GUS exon with GUSP1-GFP fusion fragment. The construct was transformed into Agrobacterium tumefaciens and transient expression assay was confirmed by agro-infiltration of Nicotiana benthamiana leaves and green fluorescence under a confocal microscope. Gene integration and expression in transgenic plants was observed through Southern blot and real-time PCR analyses. Cellular localization was observed through a confocal microscope and the copy number of the transgene was observed in progeny plants. Results: Transformation efficiency was 1.9%. Developmental and spatial expression of GUSP1 was observed through Real-time PCR in stem, root, leaf, inflorescence, and seeds of transgenic plants at the vegetative and flowering stage. Integration of GUSP1 revealed a fragment of approximately 500 bp in Southern Blot analyses. Localization of GUSP1 was detected in the intact leaf of transgenic plants through GFP fluorescence in midrib, guard cells of stomata, and trichomes. Single gene copy was detected in the chromosome of transgenic seeds. Conclusion: GUSP1 has cloned from native species of local cotton and its integration and expression in transgenic plants confirmed that the role of GUSP1 will provide direction to breed economically important cotton varieties.

Background: Cypermethrin is a Synthetic Pyrethroid (SP) having widespread applications in agriculture and industrial sector especially in sheep dip formulations and tanneries. Rhizoremediation offers a sustainable, environment-friendly and cost-effective means to carry out remediation of contaminated soils. Methods: Six bacterial strains were screened out and characterized at various doses of cypermethrin, heavy metal salts and antibiotics. The optimum growth conditions were determined for these bacterial isolates. The degradation of cypermethrin was confirmed through the growth of bacteria on minimal media (BHB) with cypermethrin and thin layer chromatographic analysis; retention factor values (Rf) were calculated and compared with standard Rf values. Results: Growth curve experiments revealed that three bacterial isolates were able to grow in the presence of cypermethrin. Tolerance to the high concentration of heavy metal salts (300µgmL-1) and resistance towards different antibiotics was observed in all three bacterial isolates indicating a positive correlation between pesticide degradation and tolerance to metals and antibiotics. Bacterial strains A-C1 and B-B2 were identified as Xanthomonas maltophilia and B-C2 as Acinetobacter sp. Cypermethrin degradation occurred concomitant with bacterial growth reaching an optical density (OD600) up to 0.869. Conclusion: Microbes present in rhizosphere have potential to mineralize the pesticides. A significant biodegradation of the cypermethrin was observed based on above mentioned lab parameters. These results paved the way for designing a multi-resistant bacterium that can be used to reverse the altered environment.

The adaptive mechanisms in Agave species enable them to survive and exhibit remarkable tolerance to abiotic stresses. Quantitative real-time PCR is a highly reliable approach for validation of targeted differential gene expression. However, stable housekeeping gene(s) is prerequisite for accurate normalization of expression data by qRT-PCR. Till date, no systematic validation study for candidate housekeeping gene identification or evaluation has been carried-out in Agave species. A total of 17 candidate housekeeping genes were identified from the de novo assembled transcriptomic data of A. sisalana and rigorously analyzed for expression stability assessment under drought, heat, cold and NaCl stress. Different statistical algorithms like geNorm, BestKeeper, NormFinder, and RefFinder on expression data determined the superior housekeeping gene(s) for accurate normalization of the gene of interest (GOI). The comprehensive evaluation revealed the β-Tub 4, WIN-1 and CYC-A as the most stable, while EEF1α, GAPDH, and UBE2 were ranked as the least stable genes in pooled samples. Pairwise combination by geNorm showed that up to two housekeeping genes would be adequate to normalize the GOI expression data precisely. Validation of identified most and least stable housekeeping genes was carried-out by normalizing the expression data of AsHSP20 under abiotic stress conditions. Copy number of AsHSP20 gene supports the reliability of the genes used for normalization. This is the first report on the screening and validation of the housekeeping genes under abiotic stress condition in A. sisalana that would assist to understand the stress tolerance mechanisms by novel gene identification and accurate validation. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s12298-020-00760-y) contains supplementary material, which is available to authorized users.

Introduction: Rocuronium is very extensively exercised nonquaternary amino steroid non-depolarizing muscle relaxant of intermediate period through the quick beginning to attain optimal situations for endotracheal intubations. The inoculation of rocuronium bromide throughout introduction of anesthesia has frequently been related by discomfort encouraged removal undertaking near place of injection. This was hypothesized that calculation of ketorolac in addition Lidocaine in grouping as pre-cure medicines amongst cases experiencing GA by rocuronium injection offers the healthier measure of removal activities in assessment to cases who obtain solitary lidocaine. The motive of our current research remained to associate effectiveness of pre-cure of mixture of ketorolac also Lidocaine in addition Lidocaine alone in occurrence of extraction undertaking related by rocuronium inoculation in peripheral veins throughout intubation. Methodology: This current research was conducted in Mayo Hospital Lahore, Pakistan from April 2018 to December 2018.Our current research remained led on 100 respondents experiencing elective operations underneath GA in OT multifaceted of the current hospital. Respondents stayed randomly separated in set-A & set-B through lottery method. Set-A established 20 mg lidocaine 4 before rocuronium. Set-B established lidocaine 23 mg in addition ketorolac 13 mg IV. GA remained managed through introduction through 7 mg/kg thiopental sodium in the distinct marginal intravenous line. Removal activities stayed detected as minor, modest in addition plain, in addition noted on the well-organized Performa. Efficiency remained defined as not any removal crusade on injecting rocuronium. Results: The variances in age in addition sex of cases remained not substantial in together sets also those remained not related through effectiveness of cure in sets. Though, ASA position of cases remained expressively related through effectiveness of cure sets. In Set-A 28(62%) in addition in Set-B 37(82%) respondents were not extraction measure whereas minor undertaking remained detected in 13(27.8%) cases in Set-A, also in 8(17.8%) cases in Set-B. Reasonable undertaking remained realized in 7(14.7%) cases in Set-A, also in 3(6.3%) cases in Set-B. The sum of cases which had removal measure stayed 19(42%) against 10(19%) in Set-A also Set-B individually (p = 0.0386). The standard of efficiency stayed satisfied through 28(57%) Set-A cases, associated to 37(81%) cases in Set-B. Conclusion: Outcomes of our existing research displayed that grouping of intravenous lidocaine also ketorolac before rocuronium inoculation remains extra actual that lidocaine unaided for averting removal activities for GA. Key words: Extraction measure; Lidocaine; Rocuronium; GA; Efficiency.

Objective: The key purpose of our existing research was to evaluate efficiency of physical treatment, spreading of Sciatic audacity to ease long-lasting discomfort in Para central disc prolapse and offer excellent attention to patient over actual cure of Para central Disc prolapse. Methods: This was a process of 6 months for 30-minute physical treatment pragmatic by widening to sciatic nerve and positional issue treatment and request of transcutanous electrical courage inspiration for 15 minutes weekly. Our current research was conducted in Lahore General Hospital from June 2018 to November 2018. Results: Our results are subsequently primary therapy, patient could a little sciatic pain for a week. As soon as pain came again, patient came to know what caused flare-up and loosened his designs of measure to decrease uneasiness. There was the reappearance of discomfort in 4th and 5th weeks owing to customary designs and work-related anxiety that needed physical treatment to medicine. From 5th week on, disc prolapsed (sciatic aching) uneasiness was seldom practiced. CONCLUSION: The current research proves efficiency of physical treatment having the positional announcement treatment to progress long-lasting sciatic aching owing to disc prolapsed. Signs by reintegration are significantly enhanced by adding of day-to-day extending package of sciatic nerve plus somatic teaching that recovers patient's consciousness of characteristic designs. Keywords: PRT submissions, backache, PIVD, nerve impingement.

The wireless transfer of electricity has gained vast importance in recent years. Many attempts had been done in this regard. The information generated is quite scattered and distributed. There is a need to harmonize the research findings of various researchers in an adept way. This review paper presents the critical evaluation of the research works done in the field of wireless power transmission since 1891. The attempt has been done in a cohesive way to connect the various findings and discussing their capabilities and limitations. The paper also highlights the major challenges faced by the research area and the new area of applications. The study closes with a conclusive review of latest scientific techniques and theoretical concepts, currently prevalent in wireless power transmission.