Multi-parametric thrombus profiling microfluidics detects intensified biomechanical thrombogenesis associated with hypertension and aging.

Arterial thrombosis, which represents a critical complication of cardiovascular diseases, is a leading cause of death and disability worldwide with no effective bioassay for clinical prediction. As a symbolic feature of arterial thrombosis, severe stenosis in the blood vessel creates a high-shear, high-gradient flow environment that effectively facilitates platelet aggregation towards vessel occlusion even with platelet amplification loops inhibited. However, no approach is currently available to comprehensively characterize the size, composition and platelet activation status of thrombi forming under this biorheological condition. Here, we present a thrombus profiling assay that monitors the multi-dimensional attributes of thrombi forming in conditions mimicking the physiological scenario of arterial thrombosis. Using this platform, we demonstrate that different receptor-ligand interactions contribute distinctively to the composition and activation status of the thrombus. Our investigation into hypertensive and older individuals reveals intensified biomechanical thrombogenesis and multi-dimensional thrombus profile abnormalities, demonstrating a direct contribution of mechanobiology to arterial thrombosis and endorsing the diagnostic potential of the assay. Furthermore, we identify the hyperactivity of GPIbα-integrin αIIbß3 mechanosensing axis as a molecular mechanism that contributes to hypertension-associated arterial thrombosis. By studying the interactions between anti-thrombotic inhibitors and hypertension, and the inter-individual variability in personal thrombus profiles, our work reveals a critical need for personalized anti-thrombotic drug selection that accommodates each patient's pathological profile.

Demographic and clinical features of dengue fever infection in Pakistan: a cross-sectional epidemiological study.

BACKGROUND: Dengue fever caused by dengue virus is a tropical disease and is among the deadliest vector-borne diseases. The humid and hot summers of Pakistan support the probation of the vectors responsible for the transmission of viral and other parasitic diseases. METHODOLOGY: A retrospective study, from 2012- 2019, of dengue infected individuals from the Punjab province of Pakistan was carried out to analyze epidemiology, clinical and laboratory findings of subjects with dengue virus infection. Data was derived from National Institute of Health (NIH) followed by Dengue control program of Pakistan, covering the incidence rate in 36 districts of Punjab and Islamabad Capital Territory (ICT) respectively. Patients data including the presence of dengue specific antigen or/and antibodies such as NS1 and IgG/IgM were observed. The study also included the analysis of demographic data, geographic data, and the month-wise distribution of dengue cases to examine seasonal trends. RESULTS: We analyzed 25,682 dengue infected individuals. The statistical analysis revealed a significant association between genders in which male population was more affected by dengue than females. It was also noted that the middle age group was the most affected age group while the highest number of cases were reported in October. Rawalpindi and Lahore were the most affected cities in Punjab province while Islamabad represented the highest number of cases during the recent outbreak in 2019. The IgM and IgG antibodies were highly prevalent among the infected patients. CONCLUSION: Dengue is endemic in Pakistan, circulating throughout the year. Highest number of cases were observed in the month of October, September and November respectively. Association between climate change and vector-borne diseases need to be investigated in Pakistan as they significantly influence the timing and intensity of dengue and other disease outbreaks. Further exploration of hematological parameters is required to better diagnose and treat the disease. For the effective control of dengue outbreaks, awareness campaigns on sewage management and vector control along with social factors are strongly recommended for better control and eradication of the disease.

The contemporary immunoassays for HIV diagnosis: a concise overview.

Recent advances in human immunodeficiency virus (HIV) diagnostics have improved the management of disease progression significantly, which have also boosted the efficacy of antiviral therapies. The detection of HIV at the earliest is very important. A highly recognized and effective virological biomarker for acute HIV infections is p24 antigen. This brief overview is based on advances of HIV diagnosis while focusing on the latest HIV testing technologies including HIV-specific antigens detecting assays of both anti-HIV antibodies and p24 antigen. In addition to other emerging molecular diagnostics for acute HIV infection, the utilization of p24 antigen has been summarized. Moreover, it has been explained how these immunoassays have reduced the window period for detection of HIV in the acute stage of infection.

Origin, Pathogenesis, Diagnosis and Treatment Options for SARS-CoV-2: A Review.

Emerging viral infections are among the greatest challenges in the public health sector in the twenty-first century. Among these, most of the viruses jump from other species of animals to humans called zoonotic viruses. The Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), by crossing species-barrier, has infected the human population for the third time in the current century and has caused the coronavirus disease-2019 (COVID-19) . Mutation and adaptation for years have greatly influenced the co-evolution and existence of coronaviruses and their possible hosts including humans. The appearance of SARS-CoV-2 in China thrust coronaviruses into the limelight and shocked the world. Presently, no coronavirus vaccines are clinically available to combat the virus's devastating effects. To counter the emergence of the COVID-19 pandemic, it is therefore important to understand the complex nature of coronaviruses and their clinical attributes. SARS and MERS outbreaks had ultimately led to socio-economic deprivation in the previous decades. In addressing the recent disastrous situation, the COVID-19 pandemic still needs some lessons from prior experience. In this review, we have highlighted the chronological order of coronavirus strains, their genomic features, the mechanism of action of SARS-CoV-2, and its disastrous repercussions on the world. We have also suggested some therapeutic options that could be effective against the COVID-19.

Cracking the Challenge of Antimicrobial Drug Resistance with CRISPR/Cas9, Nanotechnology and Other Strategies in ESKAPE Pathogens.

Antimicrobial resistance is mushrooming as a silent pandemic. It is considered among the most common priority areas identified by both national and international agencies. The global development of multidrug-resistant strains now threatens public health care improvement by introducing antibiotics against infectious agents. These strains are the product of both continuous evolution and unchecked antimicrobial usage (AMU). The ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) are the leading cause of nosocomial infections throughout the world. Most of them are now multidrug-resistant, which pose significant challenges in clinical practice. Understanding these bacteria's resistance mechanisms is crucial for developing novel antimicrobial agents or other alternative tools to fight against these pathogens. A mechanistic understanding of resistance in these pathogens would also help predict underlying or even unknown mechanisms of resistance of other emerging multidrug-resistant pathogens. Research and development to find better antibacterial drugs and research on tools like CRISPER-Cas9, vaccines, and nanoparticles for treatment of infections that can be further explored in the clinical practice health sector have recognized these alternatives as essential and highly effective tools to mitigate antimicrobial resistance. This review summarizes the known antimicrobial resistance mechanisms of ESKAPE pathogens and strategies for overcoming this resistance with an extensive overview of efforts made in this research area.

Evaluation of hematological parameters as an indicator of disease severity in Covid-19 patients: Pakistan's experience.

BACKGROUND: The severity of COVID-19 could be evaluated by examining several blood parameters mainly white blood cell (WBC) count, granulocytes, platelet, and novel hemocytometric markers neutrophils to lymphocyte ratio (NLR), platelet-to-lymphocyte (PLR), and lymphocyte to monocyte ratio (LMR). The current study was conducted to investigate alteration in blood parameters and their association with the severity and mortality of COVID-19 patients. METHODOLOGY: An observational cross-sectional study was conducted retrospectively, a total of 101 COVID-19 positive patients were examined: 52 were mild, 24 were moderate, 09 were severe, and 16 were critically diseased patients. We also recorded 16 deaths associated with the critical group. The overall mean age observed in our study was 48.94 years, where the mean age for critical individuals was 62.12 ± 14.35 years. RESULTS: A significant association between the disease severity and elevation in blood parameters were observed. The WBC's and granulocyte count were significantly increased (p value <0.001) while the mean platelet count (165.0 × 109 /L) and red blood cell volume distribution width (RDW) were decreased in the critical group (57.86%) compared to mild group's patients (177.3%) (p = 0.83). The lymphocytes count was decreased in critical patients (1.40 × 109 /L) compared to mild patients (1.92 × 109 /L) (p = 0.28). A significant association was observed in platelet-lymphocyte ratio (p < 0.001), Neutrophil-Lymphocyte ratio (p = <0.001), and Lymphocyte-Monocyte ratio (0.011). CONCLUSION: These blood parameters could be used as a suitable biomarker for the prognosis and severity of COVID-19. Evaluating novel hemograms NLR, PLR, and LMR can aid clinicians to identify potentially severe cases at early stages, initiate effective management in time, and conduct early triage which may reduce the overall mortality of COVID-19 patients.

Sofosbuvir Resistance-associated Substitutions in the Palm Domain of HCV-NS5B RNA Dependent RNA Polymerase; Study of two Sofosbuvir non-responders.

OBJECTIVE: In current study we performed sequencing of palm domain of HCV-NS5B gene, its ancestral analysis along with amino acids substitution analysis. These analysis were done to find the molecular basis of the viral resistance against Sofosbuvir drug. METHODS: Blood samples from individuals with chronic Hepatitis C infection that were resistant to Sofosbuvir were collected. The samples were processed for their molecular characterization that included RNA extraction, Complementary DNA (cDNA) synthesize, nested PCR, gel elution, Sequencing, ancestral and 3D structure analysis. RESULTS: Evolutionary analysis revealed that current study sequences (QAU-01, QAU-02) clustered with a previously studied sequence, KY971494.1. Moreover, we reports multiple novel amino acid substitutions in the palm domain of NS5B gene such as Ile116Val, Asn117Gly, Glu246Ala, Val252Ala, Glu258Gln, Cys262Leu, Ser269Arg, Ala272Thr, Ile293Leu, Lys304Arg, Asn307Gly, Ala338Val and Arg345Gly in our query sequence (QAU-01). At 246 and 269 position in (QAU-02), no substitution was observed. CONCLUSIONS: We have noticed that the current sequences are relatively emerging and could have been originated from aforementioned sequence recently. Based on the current results, we suggests that these substitutions could be associated with structural or functional impairment of protein and could also be may be considered as resistance associated substitutions (RAS) to Sofosbuvir drug.

Green fabrication of Co and Co3O4 nanoparticles and their biomedical applications: A review.

Nanotechnology is the fabrication, characterization, and potential application of various materials at the nanoscale. Over the past few decades, nanomaterials have attracted researchers from different fields because of their high surface-to-volume ratio and other unique and remarkable properties. Cobalt and cobalt oxide nanoparticles (NPs) have various biomedical applications because of their distinctive antioxidant, antimicrobial, antifungal, anticancer, larvicidal, antileishmanial, anticholinergic, wound healing, and antidiabetic properties. In addition to biomedical applications, cobalt and cobalt oxide NPs have been widely used in lithium-ion batteries, pigments and dyes, electronic thin film, capacitors, gas sensors, heterogeneous catalysis, and for environmental remediation purposes. Different chemical and physical approaches have been used to synthesize cobalt and cobalt oxide NPs; however, these methods could be associated with eco-toxicity, cost-effectiveness, high energy, and time consumption. Recently, an eco-friendly, safe, easy, and simple method has been developed by researchers, which uses biotic resources such as plant extract, microorganisms, algae, and other biomolecules such as starch and gelatin. Such biogenic cobalt and cobalt oxide NPs offer more advantages over other physicochemically synthesized methods. In this review, we have summarized the recent literature for the understanding of green synthesis of cobalt and cobalt oxide NPs, their characterization, and various biomedical applications.

Chemiluminescent-microparticle-immunoassay-based detection and prevalence of human immunodeficiency virus infection in Islamabad, Pakistan.

An advanced detection assay is important for clinical diagnosis of human immunodeficiency virus (HIV) infection in asymptomatic individuals. The first step in timely treatment and management of the infection is the early detection of HIV. In this study, we determined the prevalence of HIV in Islamabad for the first time using a chemiluminescent microparticle immunoassay (CMIA), which is an advanced serological diagnostic technique. A total of 3659 samples were obtained from the general public in Islamabad, Pakistan, and 85 of them were found to be positive for HIV infection (2.32% prevalence). Of the positive subjects, 78.82% (67/85) were male, 10.6% (9/85) were female, and 10.6% (9/85) were of unidentified gender. The results revealed a significant relationship between age groups and HIV status. The age group of 21- to 30-year-olds was found to have the highest rate of HIV infection, and the rate of HIV infection in males was higher than in females.

Impact of COVID-19 on polio vaccination in Pakistan: a concise overview.

The pandemic of coronavirus disease 2019 (COVID-19) has disrupted immunization programs around the globe, potentially increasing life-threatening vaccine-preventable diseases. Pakistan and Afghanistan are the only countries, which are still struggling to eradicate wild poliovirus. All vaccination campaigns in Pakistan were suspended in April due to the COVID-19 outbreak, leading 40 million children to miss out on polio vaccination. Like the climate crisis, the COVID-19 pandemic could be regarded as a child-rights crisis because it could have life-threatening impact over children, who need immunization, now and in the long-term. Delays in polio vaccination programs might not have immediate impact but, in the long-term, the increase in polio cases in Pakistan could result in the global export of infections. Therefore, healthcare authorities must intensify their efforts to track and vaccinate unvaccinated children in countries like Pakistan and Afghanistan. Polio vaccination campaigns need to resume immediately, so we suggest applying social distancing measures along with standard operating procedure to flatten the transmission curve of COVID-19. Furthermore, the concurrent emergence of cVDPV2 means that tOPV should temporarily be used for primary immunization. In the current review, we have discussed delays in polio vaccination, surveillance of polio viruses, reported cases in Pakistan along with recommendations to overcome interrupted immunization.

Epidemiological features of cutaneous leishmaniasis endemic in hilly areas of district Karak, Khyber-Pakhtunkhwa province of Pakistan.

Cutaneous leishmaniasis is a vector borne infection caused by flagellated parasitic protozoans of the genus Leishmania. This is an emergent threat and endemic in areas of tropic and sub-tropics and has a wider geographical distribution. This study was aimed to find the epidemiological aspects of cutaneous leishmaniasis during the recent epidemic in district Karak in 2018-2019. The suspected patients of leishmaniasis from different areas of district Karak were examined. We came across 803 cases during six-month period from August 2018 to January 2019. A major portion (> 37%; n = 299) of these patients were in age 11-20 years. The infection rate was higher in female (55.3%; n = 803) compared to males (44.6%; n = 803). Both gender and age showed a significant effect on the occurrence of infection. It is concluded that cutaneous leishmaniasis has appeared as a major health issue in district Karak. The current study report CL outbreak in Karak district which need immediate response from the healthcare authorities. In addition, extensive awareness campaigns are needed for timely prevention of such outbreaks.

Dengue Outbreaks in Khyber Pakhtunkhwa (KPK), Pakistan in 2017: An Integrated Disease Surveillance and Response System (IDSRS)-Based Report.

The current study is a retrospective epidemic report regarding dengue fever (DF) virus infection cases (2017) from fifteen districts of KPK, Pakistan. Medical records of 120 948 patients were reviewed retrospectively for demographic, clinical and laboratory data. The presence of dengue infection was confirmed by NS1-ELISA and RT-PCR, respectively. The total positive cases (of suspected DF samples) were 24 938 (20.6%), whereas seventy cases (0.28%) had a fatal outcome. Mean age ± SD of the dengue patients was 26 ± 19.8 years, while; the most affected age group was from 16 to 30 years (Chi-square: 12 820.125, p: 0.00). The infected males were 65.3%, and that of the female was 34.7%. All the dengue-infected patients were observed with symptoms of severe fever (100%), body aches (95%), gums and nose bleeding (5%), skin rashes (30%), vomiting (70%). The highest infection rate was found in district Peshawar and that of the lowest was in Bannu, Hungu and Luki Marwat. A high rate of dengue infection was found in post-monsoon months i.e. October (41%) and September (32%) of the year. The results proved that if the dengue outbreaks reveal further in KPK, it could alarmingly increase the mortality rate. Therefore, the Department of Public Health in KPK, Pakistan may take proper measures to avoid and control dengue epidemics in the future.The current study is a retrospective epidemic report regarding dengue fever (DF) virus infection cases (2017) from fifteen districts of KPK, Pakistan. Medical records of 120 948 patients were reviewed retrospectively for demographic, clinical and laboratory data. The presence of dengue infection was confirmed by NS1-ELISA and RT-PCR, respectively. The total positive cases (of suspected DF samples) were 24 938 (20.6%), whereas seventy cases (0.28%) had a fatal outcome. Mean age ± SD of the dengue patients was 26 ± 19.8 years, while; the most affected age group was from 16 to 30 years (Chi-square: 12 820.125, p: 0.00). The infected males were 65.3%, and that of the female was 34.7%. All the dengue-infected patients were observed with symptoms of severe fever (100%), body aches (95%), gums and nose bleeding (5%), skin rashes (30%), vomiting (70%). The highest infection rate was found in district Peshawar and that of the lowest was in Bannu, Hungu and Luki Marwat. A high rate of dengue infection was found in post-monsoon months i.e. October (41%) and September (32%) of the year. The results proved that if the dengue outbreaks reveal further in KPK, it could alarmingly increase the mortality rate. Therefore, the Department of Public Health in KPK, Pakistan may take proper measures to avoid and control dengue epidemics in the future.

Production of nitrogen fixing Azotobacter (SR-4) and phosphorus solubilizing Aspergillus niger and their evaluation on Lagenaria siceraria and Abelmoschus esculentus.

AIMS: Current study was aimed to produce nitrogen fixing Azotobacter sp. (SR-4) and phosphorus solubilizers Aspergillus niger (A. niger) and to evaluate their efficiency as biofertilizers for agricultural practices. METHODS: Two biofertilizer including nitrogen fixing and phosphorus solubilizing were grown. The nitrogen fixing efficiency of Azotobacter (SR-4) was determined by Kjeldahl method. Similarly, Vanadomoybdate method was used to measure the soluble phosphorus while Heinonen method was used to analyze concentration of phytase and phosphatase in the cultures. Furthermore, both biofertilizers were tested in a field trail on Lagenaria siceraria (bottle gourd) and Abelmoschus esculentus (okra). RESULTS: The Azotobacter (SR-4) strain was found efficient nitrogen fixer as 35.08 mg of nitrogen per gram of carbon was produced after 72 h of fermentation. Similarly, A. niger strain excrete extracellular phosphate solubilizing enzymes such as phytase (133UI in 48 h of fermentation) and phosphatase (170UI in 48 h of fermentation) which can solubilize the rock phosphate and make it available to plants. In field trials on selected plants (L. siceraria and A. esculentus), both biofertilizers showed significant increase in plant height, leaf length/width, fruit size and number of fruits per plant when compared with controls/untreated plants. Furthermore, plants co-inoculated with both the N fixing Azotobacter and phosphorus solubilizing A. niger have enhanced performance than those treated with each biofertilizer alone. CONCLUSION: The inoculation of seeds with A. niger and Azotobacter may replace costly and environment toxic chemical fertilizers with environment friendly and cost effective biofertilizers.