Investigating the role of Casd1 in the host response to malaria infection

Malaria is a global health concern responsible for thousands of deaths worldwide annually. Concerted efforts employing various preventive and treatment strategies have contributed a lot in controlling the disease, but we are still far from achieving complete eradication. Antimalarial drugs have been on the forefront of malaria control strategies, but their effectiveness is jeopardised due to increasing development of drug resistance in parasites. Interestingly, it has been noted that certain genetic mutations, especially those in red blood cells, confer natural resistance against malaria in endemic populations. Screening such mutations have promising translational potential as they may lead towards novel therapeutic targets or host-directed-therapies to overcome malaria resistance. In this study, N-Ethyl-N-Nitrosourea (ENU) mutagenesis, has been used as a tool to screen for novel erythrocyte variants in mouse model and assess their role in malaria susceptibility. A recent phenotypic screen for red cell abnormality identified a novel Casd1 gene mutation in mice that conferred a complete protective effect against P. chabaudi and P. berghei rodent malaria parasite. Casd1 is a eukaryotic 7(9)-O-acetyltransferase that resides in the lumen of Golgi complex as a membrane integrated protein. It functions by acetylating sialic acid residues that are subsequently decorated on the cell surface. The mutation was characterised to be a single base substitution at the splice site of Casd1 gene leading to skipping of the exon 14. An in silico analysis of the secondary structure of mutant Casd1 protein predicted a loss of connecting loop between two transmembrane helices affecting the topology of transmembrane domain. The initial screen identified a loss of Ter119 binding to erythrocytes derived from Casd1 mutated mice (ENU25^Casd1/Casd1). Ter119 is an erythroid lineage marker that recognises 7-9-di-O acetyl sialic acid epitope on the erythrocyte surface. Abrogation of Ter119 signal from the erythrocyte surface may indicate the loss of Casd1 function in ENU25^Casd1/Casd1 mice. The erythrocytes from ENU25^Casd1/Casd1 mice were further assessed for their susceptibility to invasion by malaria parasites. The relative parasitaemia was significantly reduced in the mutant erythrocytes as compared to the wild type, suggesting a novel role of Casd1 in merozoite invasion. Furthermore, the parasitised erythrocytes were tracked during infection assays to assess the role of Casd1 mutation in clearance of malaria parasites. These assays revealed a drastic clearance of exogenous blood injected into ENU25^Casd1/Casd1 mice as compared to wildtype blood. The clearance response in ENU25^Casd1/Casd1 mice was subsequently identified to be dose dependent and non-specific to malaria infection. Overall, these findings suggest that Casd1 plays a role in malaria susceptibility by modulating merozoite invasion. The results also revealed a novel role of Casd1 in self-nonself recognition which results in bystander clearance of the malaria infected foreign red blood cells in ENU25^Casd1/Casd1 mice

Predictors of Mortality of COVID-19 cases In Benazir Bhutto Hospital Rawalpindi

Background: There has been a global epidemic of COVID-19 caused by novel corona virus (SARS-2). Current research aims to study the demographic, clinical characteristics and co-morbidities in COVID-19 related deaths. Methodology: This observational (descriptive) study was conducted at BBH Rawalpindi based on data from 1st March-15th June 2020 after ethical approval. Inclusion criteria was the deceased COVID PCR positive cases (>18 years age) of both the genders. Exclusion criteria was negative PCR, doubtful diagnosis and expiry outside the hospital setting. Data was collected from hospital record and family members. Demographic details, symptoms, duration of hospital stay, co-morbidities, type of ventilatory support were documented. Data analysed by SPSS, significant p<0.05. Results: There were 54 expiries from1st March to 13th June, 42(78%) males & 12(22%) females. Mean age was 54.24+12.78 years. 76% had various comorbidities, i.e., diabetes (57%), hypertension (54%), ischemic heart disease (20%); stroke, cancer, COPD and hypothyroidism (<10% each). Most frequent cause of death was acute respiratory distress syndrome due to Covid-19. Two patients died of sepsis and multiorgan failure. 64% of patients received mechanical ventilation and 35% oxygen via non-rebreather mask. There was average 4 days on invasive mechanical ventilator. 51-60 years had longest duration of illness and hospitalization till death, while 20-30 years had the shortest.  The average mortality climbed up (25% to 57%) from April to May 2020. Conclusion: COVID-19 claims significant mortality. The risk factors for mortality being age above 50 years, male gender, co-morbidities like diabetes, hypertension, ischemic heart disease, need for mechanical ventilation upon admission and longer duration of illness. There is need to intensify the vaccination and prevention in the community keeping in mind these high-risk groups. The high-risk cases, need to be aggressively managed to reduced mortality and improve outcome. &nbsp

A situational analysis of HIV and AIDS in Pakistan

HIV (Human immunodeficiency virus) transmission has been reduced by protected sex and screening of blood products and other body fluids in the developed countries. It has been reported that Pakistan is at high risk of HIV/AIDS infection but presently the prevalence rate is considerably low. The number of reported cases of HIV/AIDS in Pakistan has been continuously increasing since 1987. By 2010 the total number of registered cases has reached to 6000 and this figure is on the rise with the passage of time. Some serious strategies must be implemented to control this deadly disease

Characterization of Hepatitis C Virus genotype 3a Hypervariable region 1 in patients achieved rapid virological response to alpha interferon and Ribavirin Combination therapy

<p>Abstract</p> <p>Background</p> <p>Hepatitis C virus roots a chronic liver disease. Currently approved treatment strategy includes administration of alpha interferon and ribavirin combined therapy for 24-48 weeks. One of the predictor of sustained virological response is an early virological response to treatment characterized as rapid response. Hyper variable region 1 (HVR1) of E2 protein is responsible for viral entry and acts as a target for neutralizing antibodies. Any mutation in this region would effect virus interaction with target cell and viral persistence.</p> <p>Methods</p> <p>Thirty one clones of six pre-treatment samples subjected to combination therapy were investigated. Three of the patients were rapid responders (R1, R2 and R3) and two were breakthrough responders (BT1 and BT2). Envelope 2 gene was amplified, cloned and sequenced. Amino acid substitution, frequency, composition and antigenic properties of HVR 1 of E2 protein were studied.</p> <p>Results</p> <p>In both rapid responders (R.R) (14 amino acid sites) and breakthrough responders (BT.R) (13 amino acid sites) half of the amino acid sites were either conserved or resistant to any physiochemical change due to amino acid substitution. It also indicated that average composition of hydrophilic and basic amino acids were comparatively lower in rapid responders than other samples affecting probable interaction of virus with target cells. A central non antigenic region was constant among the breakthrough responders but differed in length significantly among rapid responders reflecting the adaptive nature of HVR1 to the immune response.</p> <p>Conclusions</p> <p>We observed that although HVR1is quite variable region in HCV 3a patients responding differently to treatment it still maintains its physiochemical properties for its proper functioning and viability.</p

The impact of immediate breast reconstruction on the time to delivery of adjuvant therapy: the iBRA-2 study

Background: Immediate breast reconstruction (IBR) is routinely offered to improve quality-of-life for women requiring mastectomy, but there are concerns that more complex surgery may delay adjuvant oncological treatments and compromise long-term outcomes. High-quality evidence is lacking. The iBRA-2 study aimed to investigate the impact of IBR on time to adjuvant therapy. Methods: Consecutive women undergoing mastectomy ± IBR for breast cancer July–December, 2016 were included. Patient demographics, operative, oncological and complication data were collected. Time from last definitive cancer surgery to first adjuvant treatment for patients undergoing mastectomy ± IBR were compared and risk factors associated with delays explored. Results: A total of 2540 patients were recruited from 76 centres; 1008 (39.7%) underwent IBR (implant-only [n = 675, 26.6%]; pedicled flaps [n = 105,4.1%] and free-flaps [n = 228, 8.9%]). Complications requiring re-admission or re-operation were significantly more common in patients undergoing IBR than those receiving mastectomy. Adjuvant chemotherapy or radiotherapy was required by 1235 (48.6%) patients. No clinically significant differences were seen in time to adjuvant therapy between patient groups but major complications irrespective of surgery received were significantly associated with treatment delays. Conclusions: IBR does not result in clinically significant delays to adjuvant therapy, but post-operative complications are associated with treatment delays. Strategies to minimise complications, including careful patient selection, are required to improve outcomes for patients

Adding 6 months of androgen deprivation therapy to postoperative radiotherapy for prostate cancer: a comparison of short-course versus no androgen deprivation therapy in the RADICALS-HD randomised controlled trial

Background Previous evidence indicates that adjuvant, short-course androgen deprivation therapy (ADT) improves metastasis-free survival when given with primary radiotherapy for intermediate-risk and high-risk localised prostate cancer. However, the value of ADT with postoperative radiotherapy after radical prostatectomy is unclear. Methods RADICALS-HD was an international randomised controlled trial to test the efficacy of ADT used in combination with postoperative radiotherapy for prostate cancer. Key eligibility criteria were indication for radiotherapy after radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to radiotherapy alone (no ADT) or radiotherapy with 6 months of ADT (short-course ADT), using monthly subcutaneous gonadotropin-releasing hormone analogue injections, daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as distant metastasis arising from prostate cancer or death from any cause. Standard survival analysis methods were used, accounting for randomisation stratification factors. The trial had 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 80% to 86% (hazard ratio [HR] 0·67). Analyses followed the intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov, NCT00541047. Findings Between Nov 22, 2007, and June 29, 2015, 1480 patients (median age 66 years [IQR 61–69]) were randomly assigned to receive no ADT (n=737) or short-course ADT (n=743) in addition to postoperative radiotherapy at 121 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 9·0 years (IQR 7·1–10·1), metastasis-free survival events were reported for 268 participants (142 in the no ADT group and 126 in the short-course ADT group; HR 0·886 [95% CI 0·688–1·140], p=0·35). 10-year metastasis-free survival was 79·2% (95% CI 75·4–82·5) in the no ADT group and 80·4% (76·6–83·6) in the short-course ADT group. Toxicity of grade 3 or higher was reported for 121 (17%) of 737 participants in the no ADT group and 100 (14%) of 743 in the short-course ADT group (p=0·15), with no treatment-related deaths. Interpretation Metastatic disease is uncommon following postoperative bed radiotherapy after radical prostatectomy. Adding 6 months of ADT to this radiotherapy did not improve metastasis-free survival compared with no ADT. These findings do not support the use of short-course ADT with postoperative radiotherapy in this patient population

Duration of androgen deprivation therapy with postoperative radiotherapy for prostate cancer: a comparison of long-course versus short-course androgen deprivation therapy in the RADICALS-HD randomised trial

Background Previous evidence supports androgen deprivation therapy (ADT) with primary radiotherapy as initial treatment for intermediate-risk and high-risk localised prostate cancer. However, the use and optimal duration of ADT with postoperative radiotherapy after radical prostatectomy remains uncertain. Methods RADICALS-HD was a randomised controlled trial of ADT duration within the RADICALS protocol. Here, we report on the comparison of short-course versus long-course ADT. Key eligibility criteria were indication for radiotherapy after previous radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to add 6 months of ADT (short-course ADT) or 24 months of ADT (long-course ADT) to radiotherapy, using subcutaneous gonadotrophin-releasing hormone analogue (monthly in the short-course ADT group and 3-monthly in the long-course ADT group), daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as metastasis arising from prostate cancer or death from any cause. The comparison had more than 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 75% to 81% (hazard ratio [HR] 0·72). Standard time-to-event analyses were used. Analyses followed intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov , NCT00541047 . Findings Between Jan 30, 2008, and July 7, 2015, 1523 patients (median age 65 years, IQR 60–69) were randomly assigned to receive short-course ADT (n=761) or long-course ADT (n=762) in addition to postoperative radiotherapy at 138 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 8·9 years (7·0–10·0), 313 metastasis-free survival events were reported overall (174 in the short-course ADT group and 139 in the long-course ADT group; HR 0·773 [95% CI 0·612–0·975]; p=0·029). 10-year metastasis-free survival was 71·9% (95% CI 67·6–75·7) in the short-course ADT group and 78·1% (74·2–81·5) in the long-course ADT group. Toxicity of grade 3 or higher was reported for 105 (14%) of 753 participants in the short-course ADT group and 142 (19%) of 757 participants in the long-course ADT group (p=0·025), with no treatment-related deaths. Interpretation Compared with adding 6 months of ADT, adding 24 months of ADT improved metastasis-free survival in people receiving postoperative radiotherapy. For individuals who can accept the additional duration of adverse effects, long-course ADT should be offered with postoperative radiotherapy. Funding Cancer Research UK, UK Research and Innovation (formerly Medical Research Council), and Canadian Cancer Society

Performance of suspended and attached growth MBR systems in treating high strength synthetic wastewater

The performance of laboratory-scale attached growth (AG) and suspended growth (SG) membrane bioreactors (MBRs) was evaluated in treating synthetic wastewater simulating high strength domestic wastewater. This study investigated the influence of sponge suspended carriers in AG-MBR system, occupying 15% reactor volume, on the removal of chemical oxygen demand (COD), total nitrogen (TN) and total phosphorus (TP), and compared it to that of SG-MBR. Results showed that the removal efficiencies of COD, TN and TP in AG-MBR were 98%, 89% and 58%, respectively as compared to 98%, 74% and 38%, respectively in SG-MBR. Improved TN removal in AG-MBR systems was primarily based on simultaneous nitrification and denitrification (SND) process. These results infer that the presence of small bio-particles having higher microbial activity and the growth of complex biomass captured within the suspended sponge carriers resulted in improved TN and TP removal in AG-MBR.<br /

Sudden Death and Isolated Right Ventricular Noncompaction Cardiomyopathy: Report of 2 Autopsied Adult Cases

A predominantly right ventricular variant of isolated noncompaction cardiomyopathy is a potentially lethal disease entity, which only recently has become recognized in the clinical and cardiac imaging literature. There are currently few established morphologic criteria for the diagnosis other than right ventricular dilation and presence of excessive regional trabeculation. To date, there have been no autopsy reports of cases following either clinical diagnosis or sudden death. We report 2 adult cases of sudden unexpected death in which unexplained right ventricular dilation and prominent apical hypertrabeculation were the principal findings. The gross and microscopic results suggest pathological similarities between, or coexistence of, right ventricular noncompaction and arrhythmogenic right ventricular cardiomyopathies