Studies on the biased signalling of some novel delta opioid receptor agonists

The delta opioid receptor (DOR) is a G protein-coupled receptor (GPCR) which is important in the regulation of neuronal function, predominantly via coupling to heterotrimeric Gi/0 proteins. The receptor has been shown to be a potential target for the treatment of chronic pain and affective disorders. Although a large number of opioid agonists exist, their properties vary widely, at least partly due to their differential coupling to post-receptor signalling systems, a phenomenon referred to as ligand-biased signalling or functional selectivity. The aim of the current project was to examine the signalling properties of a set of established and novel DOR agonists in an attempt to identify compounds that have biased signalling profiles. It was hypothesized that DOR agonists with partial efficacy regarding β-arrestin recruitment would be less liable to induce receptor internalization and desensitization of G protein-mediated signalling than full agonists. Chinese hamster ovary (CHO) cells, stably transfected with GFP-tagged human (h)DOR and CHO-K1 and U2OS cells over-expressing hDOR were exposed to a number of novel selective DOR agonists compared with the commercially available agonists, SNC80, ADL5859 and DADLE. The compounds’ potencies and efficacies were measured in four different assay systems; 1. Inhibition of forskolin-stimulated cyclic AMP accumulation, 2. Extracellular signal-regulated kinase (ERK1/2) phosphorylation using an immunocytochemical In-cell Western (I-CW) assay, 3. β-arrestin 2 recruitment and 4. hDOR internalization. The latter two assays employed DiscoverX Enzyme Fragment Complementation technology. An attempt to develop a secreted placental alkaline phosphatase (SPAP) reporter gene assay to measure DOR-mediated cyclic AMP inhibition was not successful. All of the ligands were nearly full agonists in relation to cyclic AMP inhibition although some were less efficacious than the standard SNC80 regarding ERK1/2 activation. Their absolute potencies and rank orders of potency in inhibiting cyclic AMP and activating ERK1/2 were quite different, although both signalling systems were apparently Gi/o- protein mediated. In contrast, the agonists exhibited a full range of efficacies and potencies in both β-arrestin 2 recruitment and hDOR internalization assays and there was a significant correlation between the maximum efficacies of the compounds in the two assays. A potential relationship between β-arrestin 2 recruitment/ hDOR internalization and desensitization of agonist-induced cyclic AMP accumulation was explored. Responses to the highly arrestin-recruiting agonists SNC80 and DADLE desensitized fully after extended exposure, whereas the novel partial agonists PN6047 and OPD00003 resisted desensitization. Bias factors were calculated for the agonist set and both PN6047 and OPD00003 were found to be significantly biased towards G protein-mediated cyclic AMP inhibition. In conclusion, this study reports, for the first time, a detailed characterization of signalling bias for a set of selective DOR agonists in cells over-expressing human DORs. The findings suggest that it is potentially possible to predict wanted and unwanted properties of agonists by determining post-receptor signalling profiles in vitro which will facilitate the discovery and development of novel therapeutics based on the DOR

In-vitro anticoccidial potential of Saccharum officinarum extract against Eimeria oocysts

Present study was conducted to evaluate the effect of aqueous methanolic extract from Saccharum officinarum on the sporulation and morphology of oocysts of four Eimeria species (Eimeria tenella, E. necatrix, E. mitis, E. brunetti) of poultry. Sporulation inhibition bioassay was used to evaluate the activity of Saccharum officinarum extract (SOE) on the sporulation of coccidian oocysts. In this assay, unsporulated oocysts were exposed to six concentrations of S. officinarum in 10% dimethyl sulfoxide solution (w/v; 10, 5, 2.5, 1.25, 0.625 and 0.31%) while DMSO and potassium dichromate solution (K2Cr2O7) served as control groups. The Petri dishes were partially covered to allow the passage of oxygen and incubated at 25-29° C for 48 h, providing 60-80% humidity. The sporulation of the oocyst was confirmed by examining sporocysts under inverted microscope at 40x. Results showed anticoccidial activity of SOE against all Eimeria species as proved by its ability to inhibit the sporulation of the oocysts under laboratory conditions. Inhibition of sporulation was observed in dose dependent manner. S. officinarum extract at higher dose also damaged the normal morphology and shape of oocysts of Eimeria species

Frequency of SCF Leakage in Post-operative Patients of Tethered Spinal Cord in A Tertiary Care Hospital

Objective:  The study aimed to report the incidence of CSF leakage in patients with a tethered spinal cord, post-operatively. Materials and Methods:  A total of 75 individuals aged more than 2 years and of either gender who were hospitalized for surgery for tethered cord syndrome were included. All patients had a preoperative MRI of the spine, and those above the age of 6 had urodynamic tests. Clinical evaluations were performed until hospital release, then again at 3, 6, and 12 months. Urodynamic tests and spine MRIs were redone one year following surgery. Under general anesthesia, all patients had microscopic untethering procedures to release tethering materials and heal the thecal sac. Results:  52% of patients fall under TCS type ‘simple’, whereas, 48% of patients found with complex TCS. The frequency of CSF leakage in post-operative patients with a tethered spinal cord was found in 17.33%. No CSF leak was reported in the majority of patients (49%) patients in the age group 2-30 years. 52% of patients with simple TCS reported no CSF leak, whereas, only 13(18%) patients with complex TCS reported CSF leaks. 40% of patients reported no CSF leak who was diagnosed with complex TCS. There existed a significant relationship between the types of TCS (simple/complex) for CSF leak distribution. Conclusion:  This study concluded that the frequency of CSF leakage in post-operative patients with tethered spinal cord was found in 17.33% of patients. Keywords:  Tethered Spinal Cord (TCS), CSF Leakage, Lipomyelomeningocele

Case Report: An Extremely Rare Cause of Locally Recurrent Adenocarcinoma of the Spleen with Unknown Primary

The incidence of splenic tumour is relatively low compared to other organs. They are sometimes discovered incidentally on imaging. The majority of primary splenic tumours are diagnosed as benign. Primary malignant tumour of the spleen most commonly involves lymphosarcoma, reticulosarcoma, angiosarcoma and fibrosarcoma [1, 4]. The diagnostic algorithm to determine the nature of the splenic tumour should include series of laboratory tests, imaging studies, positron emission tomography (PET) scans as well as sometimes warranting the need for splenectomy followed by immunohistochemistry profiling. While the occurrence of primary splenic adenocarcinoma is rare, this case study reports recurrent primary splenic tumour with poorly differentiated adenocarcinoma in a patient who initially presented and investigated for anaemia of unknown cause. As to the best of our knowledge, this case has not been reported before and hence the present study provides insight into the response for standard treatment for locally recurrent adenocarcinoma of the spleen with unknown primary

Pattern of Congenital Heart Diseases in Paediatric Age Group

Background: To determine the pattern of differenttypes of congenital heart diseases, in paediatric agegroup.Methods: In this observational cross sectionalstudy cases of congenital heart defects (CHD) wereincluded, through simple random samplingirrespective of age and gender. Every patient’s dataon echocardiographic report clearly indicating hisCHD type along with age and gender was recorded.Variables of study i.e. type of CHDs, age, and genderwith relative frequencies were presented separatelygraphically.Results: Out of 298 diagnosed patients of CHD 156were males and 142 were females. Isolatedventricular septal defect found to be the mostcommon anomaly (32.6%). In combination atrialseptal defect and ventricular defect were found to bemost common with 5.4% burden rate. Out of 298patients Acyanotic CHDs were 67.1% while CyanoticCHDs were found to be 32.9%.Conclusion: Acyanotic CHDs were found to bedominant over Cyanotic CHDs with relativepercentages of 67.1% and 32.9% respectivel

Knowledge, attitudes, and practices of the general population of Pakistan regarding typhoid conjugate vaccine: findings of a cross-sectional study

Typhoid fever, a common enteric disease in Pakistan, caused by Salmonella typhi, is becoming an extended drug-resistant organism and is preventable through the typhoid conjugate vaccine (TCV). Public adherence to preventive measures is influenced by knowledge and attitude toward the vaccine. This study investigates the knowledge, attitudes, and practices of the general population of Pakistan toward TCV. The differences in mean scores and factors associated with typhoid conjugate vaccine knowledge, attitudes, and practices were investigated. A total of 918 responses were received with a mean age of 25.9 ± 9.6, 51% were women, and 59.6% had graduation-level education. The majority of them responded that vaccines prevent illness (85.3%) and decrease mortality and disability (92.6%), and typhoid could be prevented by vaccination (86.7%). In total, 77.7 and 80.8% considered TCV safe and effective, respectively. Of 389 participants with children, 53.47% had vaccinated children, according to the extended program on immunization (EPI). Higher family income has a higher odds ratio (OR) for willingness toward booster dose of TCV [crude odds ratio (COR) = 4.920, p–value <0.01; adjusted odds ratio (aOR) = 2.853, value of p <0.001], and negative attitude regarding the protective effect of TCV has less willingness toward the booster dose with statistical significance (COR = 0.388, value of p = 0.017; aOR = 0.198, value of p = 0.011). The general population of Pakistan had a good level of knowledge about the benefits of TCV, and attitude and practices are in favor of the usage of TCV. However, a few religious misconceptions are prevalent in public requiring the efforts to overcome them to promote the usage of vaccines to prevent the disease and antibiotic resistance

Acaricidal and insecticidal effects of essential oils against ectoparasites of veterinary importance

Ectoparasitism in animals has become an issue of great concern that needs to be resolved to prevent huge economic losses occurring to livestock industry all over the world. Synthetic adrugs have been playing a major role in controlling ectoparasites, but their frequent and irrational use has resulted in drug resistance to routinely used chemicals and their residual effects on food and environment. Therefore, this approach of using chemical acaricides and insecticides is losing its popularity and effectiveness in controlling ectoparasites. So, the development of alternative approaches in ectoparasite management is currently required. Among alternative protocols, plants and their essential oils have played remarkable role in controlling different ectoparasites (ticks, flies, mites, lice) of veterinary importance. Essential oils have been proved to be cheaper, more effective and safer therapeautic agents against different ectoparasites of livestock importance

Wind energy potential in Pakistan : A feasibility study in sindh province

The environment and the economy are negatively impacted by conventional energy sources, such as coal, gasoline, and other fossil fuels. Pakistan’s reliance on these resources has resulted in a catastrophic energy crisis. This has driven the government to make critical decisions such as early retail closures, power outages for the industrial sector, and an increase to two days a week vacations. Wind energy, accessible and affordable, will become a viable option for meeting Pakistan’s present and future energy demands. Approximately 3% of Pakistan’s land can produce nearly 132 GW of power with an installed capacity of 5 MW per km2. In this study, four zones (Karachi, Thatta, Badin, and Jamshoro) in Sindh province are assessed for the feasibility of wind energy generation. The installed capacity, generator types, and detailed specifications are provided for each zone. Moreover, the wind mapping of Pakistan is presented considering the four potential zones. The zones are analyzed using annual wind speed and power output considering wind data measured at 50 m height over one year. The higher mean speed is recorded at Jamshoro compared to other zones. The analysis indicates that all four sites are suitable for large-scale wind power generation due to their energy potential