Safety and efficacy of resmetirom in the treatment of patients with non-alcoholic steatohepatitis and liver fibrosis: a systematic review and meta-analysis.

Introduction: Non-alcoholic fatty liver disease (NAFLD), spanning from non-alcoholic steatohepatitis (NASH) to liver fibrosis, poses a global health challenge amid rising obesity and metabolic syndrome rates. Effective pharmacological treatments for NASH and liver fibrosis are limited. Objective: This study systematically reviews and meta-analyzes the safety and efficacy of resmetirom, a selective thyroid hormone receptor-ß agonist, in NASH and liver fibrosis treatment. By analyzing data from clinical trials, we aim to offer evidence-based recommendations for resmetirom's use in managing these conditions and identify avenues for future research. Methods: Electronic databases (PubMed, Scopus, Science Direct, Google Scholar, ClinicalTrials.gov, and Cochrane CENTRAL) were systematically searched, supplemented by manual screening of relevant sources. Only English-language randomized controlled trials were included. Data extraction, risk of bias assessment, pooled analyses, and meta-regression were performed. Results: Three randomized controlled trials involving 2231 participants were analyzed. Resmetirom demonstrated significant reductions in hepatic fat fraction [standardized mean difference (SMD) -4.61, 95% CI -6.77 to -2.44, P < 0.0001], NASH resolution without worsening fibrosis [risk ratio (RR) 2.51, 95% CI 1.74-3.64, P = 0.00001), and liver fibrosis improvement (RR 2.31, 95% CI 1.20-4.44, P = 0.01). Secondary outcomes showed significant improvements in lipid profiles, liver enzymes, and NASH biomarkers with resmetirom treatment. Meta-regression revealed associations between covariates and primary outcomes. Conclusion: Resmetirom exhibits promising efficacy in reducing hepatic fat, improving NASH resolution, and ameliorating liver fibrosis with a favorable safety profile. Further research is warranted to validate findings and optimize therapeutic strategies for NASH and liver fibrosis management.

Genome-wide DNA methylation profiles regulate distinct heat stress response in zebu (Bos indicus) and crossbred (Bos indicus × Bos taurus) cattle.

Epigenetic variations result from long-term adaptation to environmental factors. The Bos indicus (zebu) adapted to tropical conditions, whereas Bos taurus adapted to temperate conditions; hence native zebu cattle and its crossbred (B indicus × B taurus) show differences in responses to heat stress. The present study evaluated genome-wide DNA methylation profiles of these two breeds of cattle that may explain distinct heat stress responses. Physiological responses to heat stress and estimated values of Iberia heat tolerance coefficient and Benezra's coefficient of adaptability revealed better relative thermotolerance of Hariana compared to the Vrindavani cattle. Genome-wide DNA methylation patterns were different for Hariana and Vrindavani cattle. The comparison between breeds indicated the presence of 4599 significant differentially methylated CpGs with 756 hypermethylated and 3845 hypomethylated in Hariana compared to the Vrindavani cattle. Further, we found 79 genes that showed both differential methylation and differential expression that are involved in cellular stress response functions. Differential methylations in the microRNA coding sequences also revealed their functions in heat stress responses. Taken together, epigenetic differences represent the potential regulation of long-term adaptation of Hariana (B indicus) cattle to the tropical environment and relative thermotolerance.

Factor Xa inhibitors vs. warfarin in patients with Hughes syndrome: a systematic review and meta-analysis of randomized controlled trials.

Background: The optimal treatment regimen for patients with Hughes syndrome remains unclear. Therefore, the authors sought to compare the outcomes of warfarin vs. factor Xa inhibitors in patients with Hughes syndrome. Methods: MEDLINE, Embase, and Cochrane Central databases were searched for randomized controlled trials (RCTs) comparing 8 efficacy and safety of warfarin and factor Xa inhibitors in patients with Hughes syndrome. Recurrent thrombosis, all-cause mortality, stroke, adverse reactions, and bleeding were among 10 outcomes of interest. Mantel-Haenszel weighted random-effects model was used to calculate 11 relative risks (RRs) with 95% CIs. Results: The analysis included 625 patients from four RCTs and one post-hoc analysis. Meta-analysis showed a statistically non-significant difference between factor Xa inhibitors and warfarin in the recurrent thrombosis risk (arterial or venous) [RR 2.77 (95%, CI 0.79, 9.65); P=0.11, I2=50%]. Consistent results were revealed among patients with a previous history of arterial thrombosis [RR 2.76 (95% CI 0.93, 8.16); P=0.75, I2=0%], venous thrombosis [RR 1.71 (95% CI 0.60, 4.84); P=0.31, I2=15%] and patients who were triple antiphospholipid antibodies (aPL) positive [RR 4.12 (95% CI 0.46, 37.10); 21 P=0.21, I2=58%]. Factor Xa inhibitors were significantly associated with an increased risk of stroke [RR 8.51 (95% CI 2.35, 13.82); P=0.47, I2=0%]. Conclusion: Factor Xa inhibitors exhibited an increased risk of stroke among patients with Hughes syndrome. In addition, although not significant, the higher RRs among patients on factor Xa inhibitors may indicate a higher risk of thrombotic events associated with factor Xa inhibitors.

Perioperative sildenafil therapy in pulmonary hypertension associated with congenital cardiac disease: An updated meta-analysis.

To provide an updated meta-analysis to evaluate the efficacy and safety of sildenafil on pediatric patients with pulmonary hypertension (PH) associated with congenital heart disease (CHD). To assess the efficacy and safety of sildenafil, five outcomes, time duration of post-operative need for mechanical ventilation, time duration of post-operative ICU stay, length of hospitalization (LOH), the incidence of mortalities and pulmonary arterial pressure to aortic pressure ratio (PAP/AoP) were regarded as primary efficacy outcomes. Standardized mean difference (SMD) was calculated for continuous data. In comparison to the control group (CG), there was a significant decrease in the time duration of ICU stay in the sildenafil group (SG) (SMD = -0.61 [95% CI -1.17, 0.04]; P < 0.01, I2 = 85%). Length of hospitalization was assessed in the sildenafil and control groups (SMD = -0.18 [95% CI -0.67, 0.31] P = 0.05, I2 = 62%). However, there was no significant difference seen in mortality rates between the SG and CG (SMD = 0.53 [ 95% CI 0.13, 2.17] p = 0.61, I2 = 0%), in the time duration of postoperative mechanical ventilation between the SG and CG (SMD = -0.23 [95% CI -0.49, 0.03] p = 0.29, I2 = 19%), or PAP/AoP ratio between the SG and CG (SMD = -0.42 [95% CI -1.35, 0.51] P < 0.01, I2 = 90%). Based on our analysis, sildenafil has little to no effect in reducing postoperative morbidity and mortality due to PH in infants and children with CHD.

Insights into Covid-19 mortality: A comprehensive study of cardiovascular sensitivity, gender, race, and geography trends in the United States (2020-2023).

This investigation meticulously explores the evolving landscape of Covid-19-related mortality in the United States from 2020 to 2023. Leveraging the comprehensive CDC WONDER database, the study conducts a detailed analysis of age-adjusted mortality rates (AAMRs), considering various demographic and regional parameters. The identified pattern illustrates an initial surge in AAMRs from 2020 to 2021, followed by a subsequent decline until 2023. Notably, there is a discernible reduction in AAMRs for both the elderly (85 years and older) and infants (below one year). Within specific demographic segments, heightened AAMRs are observed among NH American Indian or Alaska Native individuals, men, and residents in particular states and regions. Emphasizing the significant impact of Covid-19 on cardiovascular health, the study underscores increased mortality rates associated with the cardiovascular and respiratory systems. AAMR rates were standardized per 100,000 population, providing a comparative metric. Noteworthy states with elevated AAMRs include Mississippi, Oklahoma, Kentucky, New Mexico, and Alabama, with the Southern region exhibiting the highest AAMR. The research sheds light on demographic and regional disparities in Covid-19-related mortality, calling for intensified efforts in prevention and treatment strategies. These findings, offering nuanced insights, serve as a guide for strategic public health initiatives to mitigate the multifaceted repercussions of the pandemic, especially among vulnerable populations.

Retroperitoneal liposarcoma: Case of a giant tumour inside the abdomen.

Retroperitoneal liposarcomas are rare malignant tumours that often grow rapidly in size and become symptomatic lat e in t he disease course, posing diagnostic a nd therapeutic challenges. Although abdominal imaging can rel iab ly diagnose the tumour, definitive diagnosis is only p ossib le th rough bio psy af ter surgical excision , w hich remains the primary treatment modality for these tumours. Long- term sur v ival is p rimarily determ ined throu gh histologic grade and post-resection tumour margins. We report t he case of a 43-year-o ld male patient, see n at Dr Ruth KM Pfau Hospital Karachi, who under went successful surgical excision for a well-differentiated retroperitoneal liposarcoma an d had no rad io gr aphic evidence o f local recurrence at both 3 and 12-month follow-ups.

Prefrontal cortex hemodynamic activity during a test of lower extremity functional muscle strength in children with cerebral palsy: A functional near-infrared spectroscopy study.

Children with cerebral palsy (CP) exhibit impaired motor control and significant muscle weakness due to a brain lesion. However, studies that assess the relationship between brain activity and performance on dynamic functional muscle strength assessments in CP are needed. The aim of this study was to determine the effect of a progressive lateral step-up test on prefrontal cortex (PFC) hemodynamic activity in children with CP. Fourteen ambulatory children with spastic CP (Gross Motor Function Classification System level I; 5-11 y) and 14 age- and sex-matched typically developing control children completed a progressive lateral step-up test at incremental step heights (0, 10, 15 and 20 cm) using their non-dominant lower limb. Hemodynamic activity in the PFC was assessed using non-invasive, portable functional neuroimaging (functional near-infrared spectroscopy). Children with CP completed fewer repetitions at each step height and exhibited lower PFC hemodynamic activity across step heights compared to controls. Lower PFC activation in CP was maintained after statistically controlling for the number of repetitions completed at each step height. PFC hemodynamic activity was not associated with LSUT task performance in children with CP, but a positive relationship was observed in controls at the most challenging 20 cm step height. The results suggest there is an altered PFC recruitment pattern in children with CP during a highly dynamic test of functional strength. Further studies are needed to explore the mechanisms underlying the suppressed PFC activation observed in children with CP compared to typically developing children.

A Novel Densely Packed 4 × 4 MIMO Antenna Design for UWB Wireless Applications.

In this article, a compact 4-port UWB (Ultra-Wide Band) MIMO (Multiple Input Multiple Output) antenna is proposed. A low profile FR-4 substrate is used as a dielectric material with the dimensions of 58 × 58 mm2 (0.52λ × 0.52λ) at 2.8 GHz and a standard thickness of 1.6 mm. The proposed design characterizes an impedance bandwidth starting from 2.8 to 12.1 GHz (124.1%). Each of the four elements of the proposed MIMO antenna configuration consists of a monopole antenna with PG (partial ground) that has a slot at its center. The corner of each patch (radiator) and ground slot are rounded for impedance matching. Each unit cell is in an orthogonal orientation, forming a quad-port MIMO antenna system. For reference, the partial ground of each unit cell is connected meticulously with the others. The simulated results of the proposed quad-port MIMO antenna design were configured and validated by fabrication and testing. The proposed Quad-port MIMO design has a 6.57 dBi peak gain and 97% radiation efficiency. The proposed design has good isolation below 15 dB in the lower frequency range and below 20 dB in the higher frequency range. The design has a measured ECC (Envelop Correlation Co-efficient) of 0.03 and DG (Diversity Gain) of 10 dB. The value of TARC (Total Active Reflection Coefficient) over the entire operating band is less than 10 dB. Moreover, the design maintained CCL (Channel Capacity Loss) < 0.4 bits/sec/Hz and MEG (Mean Effective Gain) < 3 dB. Based on the obtained results, the proposed design is suitable for the intended high data rate UWB wireless communication portable devices.

Breakthrough COVID-19 infections - Analyzing our experience.

Objective: There are many cases of post-vaccination COVID-19 globally. Also, literature on serum antibodies after vaccination is abundant. Our research focuses on breakthrough infections reported at our institution during the third wave of COVID-19. Methods: A total of 177 people recruited at the Indus Hospital Karachi between May to September 2021 with COVID-19 infection were divided into vaccinated, partially vaccinated, and unvaccinated cohorts. Furthermore, a subset of the vaccinated cohort was tested for anti-NP and anti-S antibodies. Results: There were 119 patients with breakthrough infection, however, 74% had mild symptoms. The antibodies against NP and S were found at a higher level in those who had a breakthrough infection in comparison to healthy vaccinated controls. Conclusion: Vaccination does not prevent disease but does confer some immunity causing less severe infection.

Mutual Coupling Reduction in MIMO DRA through Metamaterials.

A single negative metamaterial structure with hexagonal split-ring resonators (H-SRRs) is inserted within a two-port multiple-input multiple-output (MIMO) dielectric resonator antenna (DRA) in order to achieve a reduction of mutual coupling between closed multiple antenna elements. Between closed, tightly coupled, high-profile antenna elements, the single negative magnetic inclusions (H-SRRs) are embedded. By incorporating magnetic structures within antenna elements, the mutual coupling is significantly diminished. Mutual coupling reduction is attained by inserting an array of hexagonal split-ring resonators between the inter-spacing elements. An operative approach for the reduction of the mutual coupling between two × two MIMO DRAs initially operating at 5.2-GHz band is provided. To make the simulated design replica of the fabricated prototype, an air gap is introduced between the substrate, DRs, and H-SSRs. The addition of the air gap shifts the simulated results to 5.9 GHz, which closely resembles the measured values. The mutual coupling reduction is realized by integrating a meta-surface amid the two × two MIMO DRAs, which are settled in the H-plane. The meta-surface embraces an array of hexagonal split-ring resonator (H-SRR) cells that are unified along the E-plane. The H-SRR structure is designed to offer band-stop functionality within the antenna bandwidth. The proposed design has an overall dimension of 40 × 58.3 × 4.75 mm3 (1.5λ × 1.02λ × 0.079λ). By stacking the DRA with a one × three array of H-SRR unit cells, a 30 dB reduction in the mutual coupling level is attained without compromising on the antenna performance. The corresponding mutual impedance of the MIMO DRA is better than 30 dB over 5.9-6.1 GHz operating bandwidth. The proposed design has a DG of 10 db, ECC < 0.02, CCL < 0.02 bits/s/Hz, and an MEG of 0 dB. The overall design has a promising performance, which shows its suitability for the target wireless application.

System Identification and Hybrid Model Predictive Control in Personalized mHealth Interventions for Physical Activity.

The application of control systems principles in behavioral medicine includes developing interventions that can be individualized to promote healthy behaviors, such as sustained engagement in adequate levels of physical activity (PA). This paper presents the use of system identification and control engineering methods in the design of behavioral interventions through the novel formalism of a control-optimization trial (COT). The multiple stages of a COT, from experimental design in system identification through controller implementation, are illustrated using participant data from Just Walk, an intervention to promote walking behavior in sedentary adults. ARX models for individual participants are estimated using multiple estimation and validation data combinations, with the model leading to the best performance over a weighted norm being selected. This model serves as the internal model in a hybrid MPC controller formulated with three degree-of-freedom (3DoF) tuning that properly balances the requirements of physical activity interventions. Its performance in a realistic closed-loop setting is evaluated via simulation. These results serve as proof of concept for the COT approach, which is currently being evaluated with human participants in the clinical trial YourMove.

Genome-wide expression analysis reveals different heat shock responses in indigenous (Bos indicus) and crossbred (Bos indicus X Bos taurus) cattle.

Environmental heat stress in dairy cattle leads to poor health, reduced milk production and decreased reproductive efficiency. Multiple genes interact and coordinate the response to overcome the impact of heat stress. The present study identified heat shock regulated genes in the peripheral blood mononuclear cells (PBMC). Genome-wide expression patterns for cellular stress response were compared between two genetically distinct groups of cattle viz., Hariana (B. indicus) and Vrindavani (B. indicus X B. taurus). In addition to major heat shock response genes, oxidative stress and immune response genes were also found to be affected by heat stress. Heat shock proteins such as HSPH1, HSPB8, FKB4, DNAJ4 and SERPINH1 were up-regulated at higher fold change in Vrindavani compared to Hariana cattle. The oxidative stress response genes (HMOX1, BNIP3, RHOB and VEGFA) and immune response genes (FSOB, GADD45B and JUN) were up-regulated in Vrindavani whereas the same were down-regulated in Hariana cattle. The enrichment analysis of dysregulated genes revealed the biological functions and signaling pathways that were affected by heat stress. Overall, these results show distinct cellular responses to heat stress in two different genetic groups of cattle. This also highlight the long-term adaptation of B. indicus (Hariana) to tropical climate as compared to the crossbred (Vrindavani) with mixed genetic makeup (B. indicus X B. taurus).

N6-Methyladenosine RNA Modification in Host Cells Regulates Peste des Petits Ruminants Virus Replication.

N6-methyladenosine (m6A) modification is a major RNA epigenetic regulatory mechanism. The dynamics of m6A levels in viral genomic RNA and their mRNAs have been shown to have either pro- or antiviral functions, and therefore, m6A modifications influence virus-host interactions. Currently, no reports are available on the effect of m6A modifications in the genome of Peste des petits ruminants virus (PPRV). In the present study, we took PPRV as a model for nonsegmented negative-sense single-stranded RNA viruses and elucidate the role of m6A modification on viral replication. We detected m6A-modified sites in the mRNA of the virus and host cells, as well as the PPRV RNA genome. Further, it was found that the level of m6A modification in host cells alters the viral gene expression. Knockdown of the METTL3 and FTO genes (encoding the m6A RNA modification writer and eraser proteins, respectively) results in alterations of the levels of m6A RNA modifications in the host cells. Experiments using these genetically modified clones of host cells infected with PPRV revealed that both higher and lower m6A RNA modification in the host cells negatively affect PPRV replication. We found that m6A-modified viral transcripts had better stability and translation efficiency compared to the unmodified mRNA. Altogether, from these data, we conclude that the m6A modification of RNA regulates PPRV replication. These findings contribute toward a way forward for developing novel antiviral strategies against PPRV by modulating the dynamics of host m6A RNA modification. IMPORTANCE Peste des petits ruminants virus (PPRV) causes a severe disease in sheep and goats. PPRV infection is a major problem, causing significant economic losses to small ruminant farmers in regions of endemicity. N6-methyladenosine (m6A) is an important RNA modification involved in various functions, including virus-host interactions. In the present study, we used stable clones of Vero cells, having knocked down the genes encoding proteins involved in dynamic changes of the levels of m6A modification. We also used small-molecule compounds that interfere with m6A methylation. This resulted in a platform of host cells with various degrees of m6A RNA modification. The host cells with these different microenvironments were useful for studying the effect of m6A RNA modification on the expression of viral genes and viral replication. The results pinpoint the level of m6A modifications that facilitate the maximum replication of PPRV. These findings will be useful in increasing the virus titers in cultured cells needed for the economical development of the vaccine. Furthermore, the findings have guiding significance for the development of novel antiviral strategies for limiting PPRV replication in infected animals.

Integrated analysis of long-noncoding RNA and circular RNA expression in Peste-des-Petits-Ruminants Virus (PPRV) infected marmoset B lymphocyte (B95a) cells.

Peste-des-Petits-Ruminants (PPR) or goat plague is an important viral disease of sheep and goats caused by the small ruminant morbilli virus or PPR virus (PPRV). Long non coding RNAs (lncRNA) and circular RNAs (circRNA) play a pivotal role in several biological processes including regulation of virus-host interactions. The present study explored the expression of lncRNA, circRNA and their functions in PPRV infected B-lymphocyte (B95a) cells. The results revealed a total of 4531 lncRNA and 2348 circRNA expression in both mock and PPRV infected samples. Analysis of differentially expressed (DE) RNA identified 123 DE-lncRNA and 39 DE-circRNA as significantly dysregulated. Functional analysis of cis-target genes of DE-lncRNA indicated activation of TCF dependent WNT signaling and PKN1 stimulated transcription process. Interactions (sponging) of microRNA (miRNA) revealed 344 DE-lncRNA-miRNA and 93 DE-circRNA-miRNA pairs. The competing endogenous RNA (ceRNA) network of lncRNA/circRNA-miRNA-mRNA in PPRV infected B95a cells was represented by 69 ceRNA pairs. We validated the DE-circRNA by targeted amplification and sequencing of back spliced junctions (BSJs). The present study revealed a profile of lncRNA, circRNA and their potential ceRNA network in PPRV infection. The results provide insight for better understanding of PPRV-host interactions.

Effect of acute heat shock on stress gene expression and DNA methylation in zebu (Bos indicus) and crossbred (Bos indicus × Bos taurus) dairy cattle.

Environmental temperature is one of the major factors to affect health and productivity of dairy cattle. Gene expression networks within the cells and tissues coordinate stress response, metabolism, and milk production in dairy cattle. Epigenetic DNA methylations were found to mediate the effect of environment by regulating gene expression patterns. In the present study, we compared three Indian native zebu cattle, Bos indicus (Sahiwal, Tharparkar, and Hariana) and one crossbred Bos indicus × Bos taurus (Vrindavani) for stress gene expression and differences in the DNA methylation patterns. The results indicated acute heat shock to cultured PBMC affected their proliferation, stress gene expression, and DNA methylation. Interestingly, expressions of HSP70, HSP90, and STIP1 were found more pronounced in zebu cattle than the crossbred cattle. However, no significant changes were observed in global DNA methylation due to acute heat shock, even though variations were observed in the expression patterns of DNA methyltransferases (DNMT1, DNMT3a) and demethylases (TET1, TET2, and TET3) genes. The treatment 5-AzaC (5-azacitidine) that inhibit DNA methylation in proliferating PBMC caused significant increase in heat shock-induced HSP70 and STIP1 expression indicating that hypomethylation facilitated stress gene expression. Further targeted analysis DNA methylation in the promoter regions revealed no significant differences for HSP70, HSP90, and STIP1. However, there was a significant hypomethylation for BDNF in both zebu and crossbred cattle. Similarly, NR3C1 promoter region showed hypomethylation alone in crossbred cattle. Overall, the results indicated that tropically adapted zebu cattle had comparatively higher expression of stress genes than the crossbred cattle. Furthermore, DNA methylation may play a role in regulating expression of certain genes involved in stress response pathways.

A Kalman filter-based Hybrid Model Predictive Control Algorithm for Mixed Logical Dynamical Systems: Application to Optimized Interventions for Physical Activity.

Hybrid Model Predictive Control (HMPC) is presented as a decision-making tool for novel behavioral interventions to increase physical activity in sedentary adults, such as Just Walk. A broad-based HMPC formulation for mixed logical dynamical (MLD) systems relevant to problems in behavioral medicine is developed and illustrated on a representative participant model arising from the Just Walk study. The MLD model is developed based on the requirement of granting points for meeting daily step goals and categorical input variables. The algorithm features three degrees-of-freedom tuning for setpoint tracking, measured and unmeasured disturbance rejection that facilitates controller robustness; disturbance anticipation further improves performance for upcoming events such as weekends and weather forecasts. To avoid the corresponding mixed-integer quadratic problem (MIQP) from becoming infeasible, slack variables are introduced in the objective function. Simulation results indicate that the proposed HMPC scheme effectively manages hybrid dynamics, setpoint tracking, disturbance rejection, and the transition between the two phases of the intervention (initiation and maintenance) and is suitable for evaluation in clinical trials.

Effect of temperature and humidity on coronavirus infection in Pakistan.

Ongoing Coronavirus epidemic (COVID-19) identified first in Wuhan, China posed huge impact on public health and economy around the globe. Both cough and sneeze based droplets or aerosols encapsulated COVID-19 particles are responsible for airborne transmission of this virus and caused an unexpected escalation and high mortality worldwide. Current study intends to investigate the correlation of COVID-19 epidemic with meteorological parameters, particularly temperature and humidity. A data set of Epidemiological data of COVID-19 for highly infected provinces of Pakistan was collected from the official website of (https://www.covid.gov.pk/) and weather data was collected from (https://www.timeanddate.com/) during the time period of 1st March to 30th September 2020. The GrapPad prism 5 Software was used to calculate the mean and standard error of mean (SEM). In the current study the incident of daily covid cases is recorded higher in the month of June while the less number of case were reported in the month of May as compared to the other months (April, May, June, July, September and August) in the four province of Pakistan. We also find out that the incident of Covid19 were high at higher temperature (like the average temperature in the month of June 37 °C) while less cases were reported in May the average temperature was 29.5 °C. Furthermore the incident of covid cases were less reported at low humidity while more intendant with high humidity. Pearson's (r) determine the strength of the relationship between the variables. Pearson's correlation coefficient test employed for data analysis revealed that temperature average (TA) and average humidity is not a significant correlated with COVID-19 pandemic. The results obtained from the current analysis for selected parameters indirect correlation of COVID-19 transmission with temperature variation, and humidity. In the present study association of parameters is not correlated with COVID-19 pandemic, suggested need of more strict actions and control measures for highly populated cities. These findings will be helpful for health regulatory authorities and policy makers to take specific measures to combat COVID-19 epidemic in Pakistan.

A Novel Effective and Safe Vaccine for Prevention of Marek's Disease Caused by Infection with a Very Virulent Plus (vv+) Marek's Disease Virus.

Marek's disease virus (MDV) is a highly contagious alphaherpesvirus that causes rapid onset lymphoma in chickens. Marek's disease (MD) is effectively controlled using vaccination; however, MDV continues to break through vaccinal immunity, due to the emergence of highly virulent field strains. Earlier studies revealed that deletion of the meq gene from MDV resulted in an attenuated virus that protects against MD in chickens challenged with highly virulent field strains. However, the meq deleted virus retains the ability to induce significant lymphoid organ atrophy. In a different study, we found that the deletion of the vIL8 gene resulted in the loss of lymphoid organ atrophy in inoculated chickens. Here, we describe the generation of a recombinant MDV from which both meq and vIL8 genes were deleted. In vitro studies revealed that the meq and vIL8 double deletion virus replicated at levels similar to the parental very virulent plus (vv+) virus. In addition, in vivo studies showed that the double deletion mutant virus (686BAC-ΔMeqΔvIL8) conferred protection comparable to CVI988, a commercial vaccine strain, when challenged with a vv+ MDV virus, and significantly reduced lymphoid organ atrophy, when compared to meq null virus, in chickens. In conclusion, our study describes the development of a safe and effective vaccine candidate for prevention of MD in chickens.

Marek's Disease Virus Cluster 3 miRNAs Restrict Virus' Early Cytolytic Replication and Pathogenesis.

Herpesvirus-encoded microRNAs (miRNAs) have been discovered in infected cells; however, lack of a suitable animal model has hampered functional analyses of viral miRNAs in vivo. Marek's disease virus (MDV) (Gallid alphaherpesvirus 2, GaHV-2) genome contains 14 miRNA precursors, which encode 26 mature miRNAs, grouped into three clusters. In this study, the role of MDV-encoded cluster 3 miRNAs, also known as mdv1-miR-M8-M10, in pathogenesis was evaluated in chickens, the natural host of MDV. Our results show that deletion of cluster 3 miRNAs did not affect virus replication and plaque size in cell culture, but increased early cytolytic replication of MDV in chickens. We also observed that deletion of cluster 3 miRNAs resulted in significantly higher virus reactivation from peripheral blood lymphocytes. In addition, pathogenesis studies showed that deletion of cluster 3 miRNAs resulted in more severe atrophy of lymphoid organs and reduced mean death time, but did not affect the incidence of MDV-associated visceral tumors. We confirmed these results by generating a cluster 3 miRNA revertant virus in which the parental MDV phenotype was restored. To the best of our knowledge, our study provides the first evidence that MDV cluster 3 miRNAs play an important role in modulating MDV pathogenesis.