DNA-nanotechnology-based nano-architecture scaffolds based on circular strands were designed in the form of DNA-nanowires(DNA-NWs)as a polymer of DNA-triangles.Circularizing a scaffold strand(84-NT)was the critical step followed by annealing with various staple strands to make stiff DNA-triangles.Atomic force microcopy(AFM),native polyacrylamide gel electrophoresis(PAGE),UV-analysis,MTT-assay,flow cytometry,and confocal imaging were performed to assess the formulated DNA-NWs and cisplatin(CPT)loading.The AFM and confocal microscopy images revealed a uniform shape and size distribution of the DNA-NWs,with lengths ranging from 2 to 4 μm and diameters ranging from 150 to 300 nm.One sharp band at the top of the lane(500 bp level)with the loss of electrophoretic mobility during the PAGE(native)gel analysis revealed the successful fabrication of DNA-NWs.The loading efficiency of CPT ranged from 66.85％to 97.35％.MTT and flow cytometry results showed biocompatibility of the blank DNA-NWs even at 95％concentration compared with the CPT-loaded DNA-NWs.The CPT-loaded DNA-NWs exhibited enhanced apoptosis(22％)compared to the apoptosis(7％)induced by the blank DNA-NWs.The release of CPT from the DNA-NWs was sustained at＜75％for 6 h in the presence of serum,demonstrating suitability for systemic applications.The IC50 of CPT@DNA-NWs was reduced to 12.8 nM CPT,as compared with the free CPT solution exhibiting an IC50 of 51.2 nM.Confocal imaging revealed the targetability,surface binding,and slow internalization of the DNA-NWs in the scavenger-receptor-rich cancer cell line(HepG2)compared with the control cell line.

COVID-19/virology* ; Cities ; Environmental Monitoring ; Genome, Viral ; Humans ; Infection Control/methods* ; Pakistan ; RNA, Viral/genetics* ; SARS-CoV-2/genetics* ; Sewage/virology*

Cytotoxicity is a predominant biological evaluation applied to search for a suitable and non-toxic bioactive compound and to determine the biocompatibility of medical devices-related human body. The broad usage of cytotoxicity tests leads to a robust establishment of cytotoxicity assays with high sensitivity and prompt results. In vitro assays are always prioritized over in vivo due to the reproducible data, reduce numbers of animal used and easily accessible material. Compounds concentration that execute 50% of cell population is determined by calculating the IC50. According to ISO10993, cytotoxicity tests must be performed to determine the biocompatibility of medical devices that has contact with human body. This is crucial to ensure the safety of research and its clinical use. Under the recommendation of ISO10995-Part 5, three categories of tests have been documented; extract elution, direct contact and indirect contact test. Each category plays significant role depending on the nature of experiment and sample used.

The study aimed to achieve enhanced targeted cytotoxicity and cell-internalization of cisplatin-loaded deoxyribonucleic acid-nanothread (CPT-DNA-NT),mediated by scavenger receptors into HeLa cells.DNA-NT was developed with stiff-topology utilizing circular-scaffold to encapsulate CPT.Atomic force microscopy (AFM) characterization of the DNA-NT showed uniformity in the structure with a diameter of 50-150 nm and length of 300-600 nm.The successful fabrication of the DNA-NT was confirmed through native-polyacrylamide gel electrophoresis analysis,as large the molecular-weight (polymeric) DNA-NT did not split into constituting strands under applied current and voltage.The results of cell viability confirmed that blank DNA-NT had the least cytotoxicity at the highest concentration (512 nM) with a viability of 92％ as evidence of its biocompatibility for drug delivery.MTT assay showed superior cyto-toxicity of CPT-DNA-NT than that of the free CPT due to the depot release of CPT after DNA-NT inter-nalization.The DNA-NT exhibited targeted cell internalizations with the controlled intracellular release of CPT (from DNA-NT),as illustrated in confocal images.Therefore,in vitro cytotoxicity assessment through flow cytometry showed enhanced apoptosis (72.7％) with CPT-DNA-NT (compared to free CPT;64.4％).CPT-DNA-NT,being poly-anionic,showed enhanced endocytosis via scavenger receptors.

Canine babesiosis is an important tick-borne protozoal disease of dogs that poses major health problem worldwide. Farm dogs in rural areas are the companion animals, that not only watch the livestock herds but also guard the house of the owners. Each farmer keeps his companion dog to get all the services. In our study, a total of 450 blood samples of farm dogs from three different ecological zones (Southern, Central and Northern regions of the province; Punjab) of Pakistan, were collected to examine through microscopy and PCR. Examination of thin blood smears revealed an overall prevalence of 12.8% (58/450) of canine babesisal parasites. However, PCR analysis revealed 46.8% (211/450) and 7.3% (33/450) samples positive for B. gibsoni and B. vogeli, respectively. The amplicons of 671 bp and 590 bp were amplified for the detection of B. gibsoni and B. vogeli, respectively through PCR. The results of multivariate analysis showed that the occurrence of canine babesiosis is higher in the Central Punjab and younger age of the dogs, while breed and sex of the host were not significantly associated with the occurrence of the disease. Mixed infection of B. gibsoni and B. vogeli was observed only in 3 dogs each in district Kasur and Rawalpindi. Our study is the first report to observe the occurrence of canine babesiosis in rural dogs in Pakistan through PCR.

OBJECTIVES: Areca nut is widely consumed in many parts of the world, especially in South and Southeast Asia, where cardiovascular disease (CVD) is also a huge burden. Among the forms of CVD, acute coronary syndrome (ACS) is a major cause of mortality and morbidity. Research has shown areca nut chewing to be associated with diabetes, hypertension, oropharyngeal and esophageal cancers, and CVD, but little is known about mortality and re-hospitalization secondary to ACS among areca nut users and non-users. METHODS: A prospective cohort was studied to quantify the effect of areca nut chewing on patients with newly diagnosed ACS by categorizing the study population into exposed and non-exposed groups according to baseline chewing status. Cox proportional hazards models were used to examine the associations of areca nut chewing with the risk of re-hospitalization and 30-day mortality secondary to ACS. RESULTS: Of the 384 ACS patients, 49.5% (n=190) were areca users. During 1-month of follow-up, 20.3% (n=78) deaths and 25.1% (n=96) re-hospitalizations occurred. A higher risk of re-hospitalization was found (adjusted hazard ratio [aHR], 2.05; 95% confidence interval [CI], 1.29 to 3.27; p=0.002) in areca users than in non-users. Moreover, patients with severe disease were at a significantly higher risk of 30-day mortality (aHR, 2.77; 95% CI, 1.67 to 4.59; p < 0.001) and re-hospitalization (aHR, 2.72; 95% CI, 1.73 to 4.26; p < 0.001). CONCLUSIONS: The 30-day re-hospitalization rate among ACS patients was found to be significantly higher in areca users and individuals with severe disease. These findings suggest that screening for a history of areca nut chewing may help to identify patients at a high risk for re-hospitalization due to secondary events.
Acute Coronary Syndrome* ; Areca* ; Asia, Southeastern ; Cardiovascular Diseases ; Cohort Studies ; Esophageal Neoplasms ; Follow-Up Studies ; Humans ; Hypertension ; Mass Screening ; Mastication* ; Mortality* ; Nuts* ; Pakistan* ; Proportional Hazards Models ; Prospective Studies

Objective: To investigate hypocholesterolemic and hypoglycemic potential of citrus peel extract and powder using rodent experimental modeling. Methods: Considering the fact, rat feeding trial was carried out for a period of 56 d to access the prophylaxis of citrus peel flavonoids by employing normal (study I), hyperglycemic (study II) and hypercholesterolemic (study III) rats. Each study was further divided into three groups to ensure the provision of selected diets, i.e., control, functional and nutraceutical diets. Each study was further divided into three groups to ensure the provision of selected diets, i.e., control, functional and nutra-ceutical diets. Results: Declining trend for total cholesterol was observed in all studies with maximum reduction(8.55%)in rat group fed on nutraceutical diet in study III.Likewise,levels of low density lipoproteins and triglycerides reduced 11.39% and 7.89% respectively in hypercholesterolemic rats. Moreover, nutraceutical diet alleviated the sera glucose level by 8.96% in study II. Conclusions: Conclusively,inclusion of citrus peel bioflavonoids in dietary therapies is a promising strategy to modulate lipidemic and glycemic attributes without imparting any deleterious effect on hematological parameters.

Objective To investigate hypocholesterolemic and hypoglycemic potential of citrus peel extract and powder using rodent experimental modeling. Methods Considering the fact, rat feeding trial was carried out for a period of 56 d to access the prophylaxis of citrus peel flavonoids by employing normal (study I), hyperglycemic (study II) and hypercholesterolemic (study III) rats. Each study was further divided into three groups to ensure the provision of selected diets, i.e., control, functional and nutraceutical diets. Each study was further divided into three groups to ensure the provision of selected diets, i.e., control, functional and nutraceutical diets. Results Declining trend for total cholesterol was observed in all studies with maximum reduction (8.55%) in rat group fed on nutraceutical diet in study III. Likewise, levels of low density lipoproteins and triglycerides reduced 11.39% and 7.89% respectively in hypercholesterolemic rats. Moreover, nutraceutical diet alleviated the sera glucose level by 8.96% in study II. Conclusions Conclusively, inclusion of citrus peel bioflavonoids in dietary therapies is a promising strategy to modulate lipidemic and glycemic attributes without imparting any deleterious effect on hematological parameters.

Background: Various parts of Pandanus foetidus Roxb. are used as traditional medicines. However, scientific reports concerning the effect of this plant on central nervous system (CNS) depression and analgesia are unavailable. This study investigated the CNS depressant and antinociceptive effects of Pandanus foetidus leaf extracts in a rodent model. Methods: The sedative and anxiolytic activities of Pandanus foetidus extract (500 g) were tested using behavioural models of Swiss albino mice, and the analgesic activity was assessed by formalin-induced pain and tail immersion tests at 200 mg/kg body weight of the mice. The data were analysed by a one-way ANOVA, a repeated measure of ANOVA and a non-parametric test (Kruskal-Wallis test) using the SPSS software. Acute toxicity was tested using an established method. Results: Compared with the aqueous fraction, the methanol, petroleum ether and chloroform fractions of the extract exhibited a more significant (P < 0.001) reduction of locomotor activity in the mice in the open field, hole-cross, and elevated plus maze (EPM). The methanol fraction maximized the duration of sleeping time caused by the thiopental sodium induction. The extract produced a significant step-down in pain, as shown by the paw licking time in the early and late phases of the formalin test. In the tail immersion test, the chloroform fraction maximally reduced the heat-induced analgesia. The extract was found to be non toxic. Conclusion: The methanol, petroleum ether, and chloroform fractions of P. foetidus have strong CNS depressant and antinociceptive effects and thus merit further pharmaceutical studies.

Poultry farming not only provides high nutritious food but also creates employment opportunity for rural masses. Documented evidences elaborates that helminth parasitism is most deciduous problem of chickens especially in developing world. Ascaridia (A.) galli, a nematode of small intestine, has been considered as the most common and important parasite of chicken. The present study was carried out to investigate prevalence and severity of A. galli in White Leghorn layers (housing type: battery cage and deep litter, 50 each) and Fayoumi-Rhode Island Red crossbred (male and female: 50 each) flock rearing at Government Poultry Farm, Dina, Punjab, Pakistan. Two hundred faecal samples were examined by using standard parasitological and McMaster egg counting technique. The overall prevalence was 24.5% at farm, 13% in White leghorn layer (battery cage=2%, deep litter=24%) and 36% in Fayoumi-Rhode Island Red (male=34%, female=38%). It was also observed that White leghorn layer rearing in deep litter had more severe infection (EPG=1920) of A. galli compare with battery cages birds (EPG=500). Parasite prevalence was significantly related with sex (P<0.05) in Fayoumi-Rhode Island Red and male birds had less number of average parasites (0.34±0.47) as compared to females (0.38±0.490). Additionally, female birds were under serious threat of infection (EPG=2270) compared with its counterpart (EPG=1250). Given the high infection rates, particular attention should be paid to management and provision of feed supplement to White leghorn layer housing in deep litter and female bird of Fayoumi-Rhode Island Red crossbred.