RuBisCO activity assays: a simplified biochemical redox approach for in vitro quantification and an RNA sensor approach for in vivo monitoring.

BACKGROUND: Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) is the most abundant soluble protein in nature. Extensive studies have been conducted for improving its activity in photosynthesis through approaches like protein engineering. Concurrently, multiple biochemical and radiolabeling assays have been developed for determining its activity. Although these existing assays yield reliable results, they require addition of multiple external components, rendering them less convenient and expensive. Therefore, in this study, we have developed two relatively cheaper, convenient, and easily reproducible assays for quantitative and qualitative estimation of RuBisCO activity. RESULTS: We simplified a contemporary NADH based spectrophotometric RuBisCO assay by using cyanobacterial cell lysate as the source for Calvin cycle enzymes. We analyzed the influence of inorganic carbon substrates, CO2 and NaHCO3, and varying protein concentrations on RuBisCO activity. Ribulose-1,5-bisphosphate (RuBP) consumption rates for the cultures grown under 5% CO2 were 5-7 times higher than the ones grown with 20 mM NaHCO3, at different protein concentrations. The difference could be due to the impaired activity of carbonic anhydrase in the cell lysate, which is required for the conversion of HCO3- to CO2. The highest RuBisCO activity of 2.13 nmol of NAD+/ µg of Chl-a/ min was observed with 50 µg of protein and 5% CO2. Additionally, we developed a novel RNA-sensor based fluorescence assay that is based on the principle of tracking the kinetics of ATP hydrolysis to ADP during the conversion of 3-phosphoglycerate (3-PG) to 1,3-bisphosphoglycerate (1,3-BPG) in the Calvin cycle. Under in vitro conditions, the fluorometric assay exhibited  ~ 3.4-fold slower reaction rate (0.37 min-1) than the biochemical assay when using 5% CO2. We also confirmed the in vivo application of this assay, where increase in the fluorescence was observed with the recombinant strain of Synechocystis sp. PCC 6803 (SSL142) expressing the ADP-specific RNA sensor, compared to the WT. In addition, SSL142 exhibited three-fold higher fluorescence when supplemented with 20 mM NaHCO3 as compared to the cells that were grown without NaHCO3 supplementation. CONCLUSIONS: Overall, we have developed a simplified biochemical assay for monitoring RuBisCO activity and demonstrated that it can provide reliable results as compared to the prior literature. Furthermore, the biochemical assay using 5% CO2 (100% relative activity) provided faster RuBP consumption rate compared to the biochemical assay utilizing 20 mM NaHCO3 (30.70% relative activity) and the in vitro fluorometric assay using 5% CO2 (29.64% relative activity). Therefore, the absorbance-based biochemical assay using 5% CO2 or higher would be suitable for in vitro quantification of the RuBisCO activity. On the other hand, the RNA-sensor based in vivo fluorometric assay can be applied for qualitative analysis and be used for high-throughput screening of RuBisCO variants. As RuBisCO is an enzyme shared amongst all the photoautotrophs, the assays developed in this study can easily be extended for analyzing the RuBisCO activities even in microalgae and higher plants.

Development of pH-Responsive, Thermosensitive, Antibacterial, and Anticancer CS/PVA/Graphene Blended Hydrogels for Controlled Drug Delivery.

Drug delivery techniques based on polymers have been investigated for their potential to improve drug solubility, reduce systemic side effects, and controlled and targeted administration at infection site. In this study, we developed a co-polymeric hydrogel composed of graphene sheets (GNS), polyvinyl alcohol (PVA), and chitosan (CS) that is loaded with methotrexate (MTX) for in vitro liver cancer treatment. Fourier transform infrared spectroscopy (FTIR) and atomic force microscopy (AFM) was employed to check the structural properties and surface morphology. Moreover, tests were conducted on the cytotoxicity, hemolytic activity, release kinetics, swelling behaviour and degradation of hydrogels. A controlled release of drug from hydrogel in PBS at pH 7.4 was examined using release kinetics. Maximal drug release in six hours was 97.34%. The prepared hydrogels did not encourage the HepG2 growth and were non-hemolytic. The current study highlights the potential of GNS-based hydrogel loaded with MTX as an encouraging therapy for hepatocellular carcinoma. HepG2 cell viability of MTX-loaded CS-PVA-GNS hydrogel was (IC50 5.87 µg/200 mL) in comparison to free MTX (IC50 5.03 µg/200 mL). These outcomes recommend that hydrogels with GNS ensure improved drug delivery in cancer microenvironment while lessening adverse consequences on healthy cells.

Elucidating the redox activity of cobalt-1,2,3,4-cyclopentane-tetracarboxylic acid and 1,2,4,5-benzene-tetracarboxylic acid-based metal-organic frameworks for a hybrid supercapacitor.

The development of electrode materials with extraordinary energy densities or high power densities has experienced a spectacular upsurge because of significant advances in energy storage technology. In recent years, the family of metal-organic frameworks (MOFs) has become an essential contender for electrode materials. Herein, two cobalt-based MOFs are synthesized with distinct linkers named 1,2,4,5-benzene-tetra-carboxylic acid (BTCA) and 1,2,3,4-cyclopentane-tetracarboxylic acid (CPTC). Investigations have been rigorously conducted to fully understand the effect of linkers on the electrochemical properties of Co-based MOFs. The best sample among the MOFs was used with activated carbon to create a battery-supercapacitor hybrid device. Due to its noteworthy results, specific capacity (100.3 C g-1), energy density (23 W h kg-1), power density (3400 W kg-1) and with the lowest ESR value of 0.4 Ω as well as its 95.4% capacity retention, the fabricated hybrid device was discovered to be very appealing for applications demanding energy storage. An approach for evaluating battery-supercapacitors was employed by quantifying the capacitive and diffusive contributions using Dunn's model to reflect the bulk and surface processes occurring during charge storage. This study fills the gap between supercapacitors and batteries, as well as providing a roadmap for creating a new generation of energy storage technologies with improved features.

Antimicrobial susceptibility and clinical characteristics of multidrug-resistant polymicrobial infections in Pakistan, a retrospective study 2019-2021.

Background: We determined the prevalence of antimicrobial resistance (AMR) in polymicrobial pathogens in Pakistan. Methods: A total of 70,518 clinical samples were collected aseptically and confirmation of isolates and antibiogram were performed by the VITEK 2 system. Results: Of 70,518 samples, 441 (0.62%) were polymicrobial samples, with 882 (1.2%) polymicrobial pathogens with 689 (78.1%) Gram-negative rods (GNRs), 166 (18.8%) Gram-positive cocci and 27 (3.1%) Candida albicans. Among GNRs, 28.8% were Escherichia coli and 25.9% were Klebsiella pneumoniae. Majority, 15.1% of Pseudomonas aeruginosa and K. pneumoniae were found in combination. 30.1% of isolates were ESBL producers, 9.7% carbapenem-resistant organisms, 35.5% MRSA and 6.0% VRE. 100% of E. coli were resistant to ampicillin and 98% of K. pneumoniae were resistant to piperacillin. Conclusion: A high prevalence of AMR in polymicrobial pathogens was observed.

Infections caused by one or more types of bacteria, viruses, fungi or parasites ­ known as polymicrobial infections ­ are a threat to health. These infections cause serious illness and are linked to high numbers of deaths, long hospital stays and high costs of treatment. Usually, polymicrobial infections are treated with combinations of antimicrobials. However, microbes becoming less susceptible to antimicrobials (known as antimicrobial resistance) is an increasing problem. To find out how common resistance is in Pakistan, this study tested 70,518 clinical samples. Of these, 441 tested positive for polymicrobial infections. These included Candida albicans, Gram-positive cocci and Gram-negative rods infections. Many of these were resistant to widely used antibiotics such as penicillins, cephalosporins, quinolones and fluoroquinolones. This study concluded that hospitals in Pakistan have a high prevalence of resistance and that better cleanliness practices should be put in place to combat this.

Purification and characterization of novel isoforms of the polyphenol oxidase from Malus domestica fruit pulp.

Polyphenol oxidases (PPOs), belong to the group of oxidoreductases that are copper containing enzymes and are responsible for plant browning. PPOs are extensively distributed in plant kingdom and can oxidize wide range of aromatic compounds of industrial importance. The aim of this study was purification and characterization of PPO isoforms from the fruit pulp of Golden delicious apple. High performance liquid chromatography was used to purify the two novel isoforms of PPO and further their molecular weights (45 and 28 kDa) were determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified isoforms have optimum pH (6.5), optimum temperature (40°C), the Vmax (4.45 µM/min) and Km (74.21 mM) with catechol substrate. The N-terminal microsequences of both PPO isoforms were determined using a pulse liquid protein sequencer and found to be AKITFHG (28 kDa) and APGGG (45 kDa). Polyphenol oxidases are efficiently used in the pharmaceutical, paper and pulp, textiles and food industries. Recently, the PPOs have been used for bioremediation and in the development of biosensors.

Transition metal dichalcogenide electrodes with interface engineering for high-performance hybrid supercapacitors.

Transition metal dichalcogenides (TMDCs) have been explored in recent years to utilize in electronics due to their remarkable properties. This study reports the enhanced energy storage performance of tungsten disulfide (WS2) by introducing the conductive interfacial layer of Ag between the substrate and active material (WS2). The interfacial layers and WS2 were deposited through a binder free method of magnetron sputtering and three different prepared samples (WS2 and Ag-WS2) were scrutinize via electrochemical measurements. A hybrid supercapacitor was fabricated using Ag-WS2 and activated carbon (AC) since Ag-WS2 was observed to be the most proficient of all three samples. The Ag-WS2//AC devices have attained a specific capacity (Qs) of 224 C g-1, while delivering the maximum specific energy (Es) and specific power (Ps) of 50 W h kg-1 and 4003 W kg-1, respectively. The device was found to be stable enough as it retains 89% capacity and 97% coulombic efficiency after 1000 cycles. Additionally, the capacitive and diffusive currents were obtained through Dunn's model to observe the underlying charging phenomenon at each scan rate.

Antioxidant and hypoglycemic potential of phytogenic cerium oxide nanoparticles.

Plants provide humans with more than just food and shelter; they are also a major source of medications. The purpose of this research was to investigate the antioxidant and hypoglycemic potential of green synthesized CeONPs using Mentha royleana leaves extract. The morphological and physicochemical features of CeONPs were evaluated by UV-Visible spectrophotometry, Scanning Electron Microscopy, Energy Dispersive X-rays and Fourier-transform infrared spectrometry, Dynamic light scattering, Atomic Force Microscopy, Zeta Potential. The average size range of synthesized CeONPs diameter between 46 and 56 nm, crystalline in shape, with Polydispersity index value of 0.2 and subatomic particles mean diameter was 4.5-9.1 nm. The antioxidant capability of CeONPs was assessed using DPPH, ABTS+, hydrogen peroxide, hydroxyl radical scavenging, and reducing power tests. The hypoglycemic potential of CeONPs was investigated using alpha-amylase, alpha-glucosidase, glucose absorption by yeast cells, and antisucrase. The effective concentrations were 500 and 1000 µg/ml found good in suppressing radical species. To explore the hypoglycemic potential of CeONPs, alpha-amylase, alpha-glucosidase, glucose absorption by yeast cell, and antisucrase assays were performed. Glucose absorb by yeast cells assay was tested for three distinct glucose concentrations: 5 mmol/L, 10 mmol/L, and 25 mmol/L. Green synthesize CeONPs showed a dose-dependent response, higher concentrations of CeONPs imposed a stronger inhibitory impact on the catalytic site of enzymes. This study suggest that CeONPs could possibly binds to the charge carrying species and act as competitive inhibitor which slow down the enzyme substrate reaction and prevents enzymatic degradation. The study's findings were outstanding, which bodes well for future medicinal applications of CeONPs.

Exploring MOF-199 composites as redox-active materials for hybrid battery-supercapacitor devices.

Metal-organic frameworks (MOFs) have emerged as intriguing porous materials with diverse potential applications. Herein, we synthesized a copper-based MOF (MOF-199) and investigated its use in energy storage applications. Methods were adapted to intensify the electrochemical characteristics of MOF-199 by preparing composites with graphene and polyaniline (PANI). The specific capacity of the synthesized MOF in a three-electrode assembly was significantly enhanced from 88 C g-1 to 475 C g-1 and 766 C g-1 with the addition of graphene and polyaniline (PANI), respectively. Due to the superior performance of (MOF-199)/PANI, a hybrid supercapacitor was fabricated with the structure of (MOF-199)/PANI//activated carbon, which displayed an excellent maximum energy and power density of 64 W h kg-1 and 7200 W kg-1, respectively. The hybrid device exhibited an appreciable capacity retention of 92% after 1000 charge-discharge cycles. Moreover, using Dunn's model, the capacitive and diffusive contributions as well as the k 1 and k 2 currents of the fabricated device were calculated, validating the hybrid nature of the supercapattery device. The current studies showed that MOF-199 exhibits promising electrochemical features and can be considered as potential electrode material for hybrid energy storage devices.

Antecedent infections, recent developments and future directions in Guillain-Barré syndrome.

The Guillain-Barré syndrome is an autoimmune polyradiculoneuropathy causing symmetrical weakness of limbs. After poliomyelitis, it is the second most common cause of paralysis, with an annual incidence of 0.84-1.91 per 100,000 individuals. The syndrome affects both men and women, showing a male preponderance. Campylobacter jejuni, epstein-barr virus, cytomegalovirus, mycoplasma pneumoniae and haemophilus influenzae are amongst the most common causative agents of Guillain-Barré syndrome. Several immunological and genetic factors have been recognised as the risk factors. Human leukocyte antigen, cluster of differentiation 1, and tumour necrosis factor-alpha alleles are among the frequently investigated loci in Guillain-Barré syndrome. Genome-wide association studies have found no significant association of Guillain-Barré syndrome with common variants. Many vaccines against Campylobacter jejuni infection have been proposed, but there are concerns about the efficacy and safety of these vaccines. So far, there is no approved vaccine against Campylobacter jejuni.

Study of Guillain-Barre syndrome etiology in Pakistani patients.

OBJECTIVE: To examine clinical features, biochemical markers, demographic features, antecedent infections, frequency and treatment strategies related to Guillain-Barré syndrome. METHODS: The case-control study was conducted at the Pakistan Institute of Medical Sciences, Islamabad, Pakistan, and the District Headquarters Hospital, Rawalpindi, Pakistan, from 2018 to 2020, and comprised Guillain-Barré syndrome patients in group A and healthy controls in group B. The patients were diagnosed on the basis of clinical presentation, nerve conduction study, electromyography, cerebrospinal fluid analysis and biochemical profile. Data was analysed using SPSS 23. RESULTS: Of the 167 subjects, 90(54%) were in group A and 77(46%) were in group B. The mean age of group A was 40.20±14.90 years, while there were 61(67.7%) males and 29(32.2%) females compared to 50 (64.93%) males and 27 (35.06%) females with mean age 38.40±12.34 years in group B. Acute inflammatory demyelinating polyneuropathy was the most common electrophysiological variant of Guillain-Barré syndrome 41(46%). There was significant difference in mean interleukin-17 levels between group A 23.12±3.41 pg/ml and group B 8.82±2.49 (p<0.05). Gastrointestinal infection was the most common preceding infection 51(56.66%). The mean cerebrospinal fluid protein was 100.83±51.32g/dl and albumiocytologic dissociation was found in all the four variants (p= 0.005). CONCLUSION: Guillain-Barré syndrome affected patients regardless of age, while males were more affected than females. Majority of the patients had an antecedent infection before disease onset. Increased levels of interleukin-17 showed involvement of autoimmunity. Albuminocytologic dissociation differentiated it from poliomyelitis.

Field Evolved Resistance in Earias vittella (Lepidoptera: Noctuidae) From Punjab, Pakistan Against Commercial Formulations of Bacillus thuringiensis kurstaki.

Transgenic commercial cotton expressing Bacillus thuringiensis (Bt) Cry endotoxins or vegetative Vip toxins provide protection to cotton against bollworm attack. Continuous exposure of these targeted pests to cry toxins and to Bt commercial spray formulations has resulted in the development of resistance through natural selection. Spotted bollworm Earias vittella (Noctuidae: Lepidoptera) is considered to be one of the most destructive pests of cotton and okra crops in South Asia including Pakistan and has developed resistance to various synthetic insecticides. In the present study, the level of resistance in field populations of the spotted bollworm E. vittella against Bt Cry toxins has been evaluated for the first time. We collected twelve populations of E. vittella from three districts of Punjab, Pakistan for testing against four commercial Bt formulations containing different strains of B. thuringiensis subspecies kurstaki (Btk) with a range of Cry toxins. Low to high levels of resistance were found in the field populations compared with a laboratory-reared susceptible population of E. vittella (resistance ratios 6 to 111-fold). These results suggest that E. vittella has developed resistance against different Cry toxins after continuous exposure to Bt cotton in field. In order to prevent field control failures, regular insecticide resistance monitoring programs are required together with the use of integrated management approaches, including the use of Bt cotton varieties expressing two or more toxins to delay the development of resistance against Bt toxins in E. vittella.

Low Effectiveness of Prosulfocarb and Mesosulfuron-Methyl + Iodosulfuron-Methyl against Vulpia myuros.

Due to natural tolerance to most widely used herbicides for grass weed control, prosulfocarb as pre-emergence or early post-emergence herbicide and mesosulfuron + iodosulfuron as post-emergence herbicide are the mainstays of any chemical control program for Vulpia myuros in Denmark. However, farmers often report variable efficacy of these herbicides on V. myuros compared to other grass weeds. Dose-response experiments were conducted to evaluate the performance of prosulfocarb and mesosulfuron + iodosulfuron on V. myuros. Prosulfocarb was sprayed at different plant growth stages to study the influence of plant growth stage on the performance of prosulfocarb on V. myuros in comparison with the more susceptible grass weed species Apera spica-venti. Doses causing 50% reduction in response variable (ED50) were estimated from the dose-response analysis. The ED50 values revealed a higher tolerance of V. myuros to prosulfocarb and mesosulfuron + iodosulfuron than A. spica-venti. The relative difference in the effectiveness of prosulfocarb between V. myuros and A. spica-venti was constant among plant growth stages studied. The highest levels of V. myuros control were achieved when prosulfocarb was sprayed pre-emergence (BBCH 00), while the control substantially declined at later growth stages. The results from the current study document the tolerance of V. myuros to prosulfocarb and mesosulfuron + iodosulfuron and highlight the importance of optimization of prosulfocarb spray timing for achieving maximum control of V. myuros.

Exploring genetic diversity and population structure of Punjab goat breeds using Illumina 50 K SNP bead chip.

Pakistan has 35 goat breeds. Moreover, the province of Punjab has highest goat population constituting 37% of country's total population with seven goat breeds including Beetal, Daira Deen Panah, Nachi, Barbari, Teddi, Pahari, and Pothwari. The diversity study of breeds warrants the documentation of breeds particularly using genome wide panel of markers, i.e., SNP chip. The objective of the current study was to fill this gap of information. Therefore, in current study we collected total of 879 unrelated goat blood samples along with data on body weight measurements; genomic DNA was extracted, and genotyping was carried out using 50 K SNP bead chip. Quality control measures were performed in Plink 1.07. Genetic diversity was observed among studied populations using heterozygosity and pairwise FST estimates, principal component analysis, admixture analysis in Plink software with visualization in Clumpak, and constructing phylogenetic tree in Mega 7 software. Moderate to high level of heterozygosity was observed among the studied populations. Coefficient of inbreeding varied from 0.0186 ± 0.0327 in Pahari to 0.183 ± 0.0715 in Barbari. Barbari and Daira Deen Panah had quite higher level of inbreeding coefficient as compared to all other breeds with value of 0.183 ± 0.0715 and 0.1378 ± 0.0741, respectively. PCA identified three steps of subdividing the seven goat breeds at various levels of K. All the seven breeds made independent clusters at various levels of PCA. Admixture analysis revealed the distinctness of Teddi and Barbari breeds. Genetic sub-structuring was observed in the admixture patterns of Beetal breed. Moreover, high level of genetic admixture was observed in Nachi, Pahari, Pothwari, and Daira Deen Panah breeds. Admixture results were further interpreted by calculating pairwise FST values. Our results provided first insights about genetic diversity of Pakistani goat breeds based on genomic data. To conclude, the enriched goat breed diversity in Pakistan could provide valuable genetic reservoir for national breeding schemes.

Preclinical Study on the Hepatoprotective Effect of Pollen Extract of Pinus brutia Ten. (Red Pine) in Mice and Phenolic Acid Analysis

Objectives: Many agents, including those from herbal sources, have been sought as preventives or cures for hepatotoxicity. The pollen of Pinus brutia Ten., known as red pine (Pinaceae), is used against liver diseases in Anatolian folk medicine. Materials and Methods: In the current study, pollen ethanol extract of P. brutia was investigated for its possible hepatoprotective activity using a mouse model of CCl4-induced hepatotoxicity. Swiss albino mice were divided into five groups, and extract-treated groups were compared with a silymarin-treated group as the reference. The extract was tested at 100, 200, and 300 mg/kg (b.w.). Phenolic acids were analyzed using high-performance column chromatography (HPLC) in the extracts as pollens are usually known to be rich in phenolics. Results: Our data revealed that the extract displayed the best hepatoprotection at a dose of 100 mg/kg when compared with silymarin (Legalon®), the reference drug. HPLC analysis indicated presence of protocatechuic acid (0.176 mg/g extract), p-hydroxybenzoic acid (0.001 mg/g extract), vanillic acid (VA) (0.537 mg/g extract), syringic acid (0.050 mg/g extract), and tr-cinnamic acid (0.310 mg/g extract), while the major phenolic acid was VA. Conclusion: The outcomes of this study allow us to conclude that red pine pollen extract can serve as a promising hepatoprotective agent. Among the phenolic acids analyzed in the pollen extract, vanillic acid as the major one besides some other phenolic acids detected seems to be responsible for its remarkable hepatoprotective effect.

Correlation of C-reactive protein levels, gene polymorphism and platelets count in Dengue infection.

OBJECTIVE: To determine the correlation of polymorphism in C-reactive protein gene with variation in serum levels in dengue patients. METHODS: The cross-sectional study was conducted at Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, Pakistan, from October 2017 to October 2018, and comprised blood samples from dengue patients which were used to measure the serum levels of C-reactive protein. Deoxyribonucleic acid extraction followed by tetra amplification-refractory mutation system polymerase chain reaction was used to analyse the genotype variation T>G for single nucleotide polymorphism rs199953854 using allele-specific primers. Correlation of serum C-reactive protein levels with the C-reactive protein polymorphism in dengue patients was explored. Data was analysed using SPSS 21. RESULTS: Of the 200 patients, 108(54%) had very high C-reactive protein levels, 48(24%) had levels slightly higher than the upper limit, 14(7%) had low and 30(15%) had normal levels. Also, 162(81%) patients had low platelets count. Amplification of only T alleles was noted. CONCLUSIONS: C-reactive protein levels were found to be increased with suppressed platelets count in dengue patients. Single nucleotide polymorphism rs199953854 appeared to have no polymorphism.

Association of ABO blood group with COVID-19 severity, acute phase reactants and mortality

Background and objectiveThe ABO blood group system has been associated with infectious and noninfectious disease, including dengue, hepatitis B virus (HBV), and severe respiratory syndrome coronavirus (SARS), etc. Coronavirus disease 2019 (COVID-19) is the ongoing pandemic with multitude of manifestations and association of ABO blood group in South-East Asian population needs to be explored. MethodsIt was a retrospective study of patients with real time polymerase chain reaction (RT-PCR) diagnosis of COVID-19 at Advanced Diagnostics and Liver Center between April 2020 to January 2021. Blood group A, B, O, and AB were identified in every participant, irrespective of their RH type and allotted groups 1, 2,3, and 4, respectively. Cox regression and logistic regression were used for inferential statistics. ResultsThe cohort included 1067 patients: 521 (48.8%) of blood group O, 295 (27.6%) of blood group B, 202 (18.9%) of blood group A, and 49 (4.5%) of blood group AB. The majority of the patients were males 712 (66.7%) with an average body mass index (BMI) of 27.45 {+/-} 3.53. Patients with AB blood group stayed a median (IQR) of 14 (5, 27) days while A blood group cohort stayed 13 (6,27) days and overall 10.6% COVID-19-related mortality was observed at our center, with 13.9% in blood group A as the majority of COVID-19 deaths. Regarding severity of COVID-19 disease, there was a trend towards critical disease in blood group A and O (n=83, 41.1%; n=183, 35.1%; OR, 11.34 (95% CI, 46.79-53.22); p<0.001). Logistic regression demonstrates blood group O and AB as predictors for severe COVID-19 disease (O: OR: 0.438 (95% CI: 0.168-1.139) p=0.090; AB: OR: 0.415 (95% CI: 0.165-1.046) p=0.062) and cause-specific hazards ratio (HR) for survival function was 3.206 (p=0.361) among all blood groups. ConclusionAlthough the prevalence of blood group O was higher in this cohort, hospital stay, severity of disease, and mortality were associated with blood group A. Further studies are needed for understanding the underlying mechanism behind the association of blood groups with COVID-19.

Effect of COVID-19 on Lipid Profile and its Correlation with Acute Phase Reactants

Background and ObjectiveCoronavirus disease 2019 (COVID-19) manifests as multiple clinical and pathological organ dysfunctions. It also disrupts metabolic profile due to the release of pro-inflammatory cytokines causing a systemic inflammation reaction. However, the development and correlation of dyslipidemia with acute phase reactants is unknown. This investigation was performed to assess the pathological alterations of low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein (HDL), triglycerides, and total cholesterol levels in COVID-19 patients. MethodsThis was a prospective study performed on real-world patients to assess serum levels of LDL-C, HDL, TG, TC on COVID-19 patients (mild: 319; moderate: 391; critical: 357) hospitalized at our center between April 2020 through January 2021. Age- and gender-matched controls who had their lipid profiles in the same period were included as the control group. ResultsLDL-C, HDL, TG, and TC levels were significantly lower in COVID-19 patients when compared with the control group (P < 0.001, 0.047, 0.045, < 0.001, respectively). All parameters decreased gradually with COVID-19 disease severity (LDL-C: median (IQR), mild: 98 (91,134); moderate: 97 (81,113); critical: 68 (68,83); HDL: mild: 45 (37,50); moderate: 46 (41,50); critical: 40 (37,46); TG: mild: 186 (150,245); moderate: 156 (109,198); critical: 111 (98,154); TC: mild: 224 (212,238); moderate: 212 (203,213); critical: 154 (125,187)). LDL-C, TC, and TG were inversely correlated with acute phase reactants (interleukin-6 (IL-6), Procalcitonin, C-reactive protein (CRP), and D-dimers). Logistic regression demonstrated lipid profile, thyroid profile, and acute phase reactants as predictors of severity of COVID-19 disease. ConclusionHypolipidemia develops in increasing frequency with severe COVID-19 disease. It inversely correlates with levels of acute-phase reactants, indicating SARS-COV-2 as the causative agent for alteration in lipid and thyroid levels.

In Silico Validation, Fabrication and Evaluation of Nano-Liposomes of Bistorta amplexicaulis Extract for Improved Anticancer Activity Against Hepatoma Cell Line (HepG2).

BACKGROUND: Bistorta amplexicaulis of the genus Polygonum (Polygonaceae) has been reported for its antioxidant and anticancer activities. However, the low cellular uptake of the compounds in its extract limits its therapeutic application. OBJECTIVES: The present study aimed at developing a nanoliposomal carrier system for B. amplexicaulis extracts for improved cellular uptake, thus resulting in enhanced anticancer activity. METHODS: Ultra Pressure Liquid Chromatography (UPLC) was used to identify major compounds in the plant extract. Nanoliposomes (NLs) were prepared by employing a thin-film rehydration method using DPPC, PEG2000DSPE and cholesterol, followed by characterization through several parameters. In vitro screening was performed against breast cancer cell line (MCF-7) and Hepatocellular carcinoma cell line (HepG-2) using MTT-assay. Raw extract and nanoliposomes were tested on Human Umbilical Vein Endothelial Cells (HUVEC). Moreover, molecular docking was performed to validate the data obtained through wet lab. RESULTS: The UHPLC method identified gallic acid, caffeic acid, chlorogenic acid and catechin as the major compounds in the extract. The NLs with a size ranging between 140-155 nm, zeta potential -16.9 to -19.8 mV and good polydispersity index of < 0.1 were prepared, with a high encapsulation efficiency of 81%. The MTT assay showed significantly (p > 0.05) high uptake and cytotoxicity of NLs as compared to the plant extract. Moreover, reduced toxicity against HUVEC cells was observed as compared to the extract. Also, the docking of identified compounds suggested a favorable interaction with the SH2 domain of both STAT3 and STAT5. CONCLUSION: Overall, the results suggest NLs as a potential platform that could be developed for the improved intracellular delivery of plant extract, thus increasing the therapeutic outcomes.

Large Area Emission in p-Type Polymer-Based Light-Emitting Field-Effect Transistors by Incorporating Charge Injection Interlayers.

Organic light-emitting field-effect transistors (LEFETs) provide the possibility of simplifying the display pixilation design as they integrate the drive-transistor and the light emission in a single architecture. However, in p-type LEFETs, simultaneously achieving higher external quantum efficiency (EQE) at higher brightness, larger and stable emission area, and high switching speed are the limiting factors for to realise their applications. Herein, we present a p-type polymer heterostructure-based LEFET architecture with electron and hole injection interlayers to improve the charge injection into the light-emitting layer, which leads to better recombination. This device structure provides access to hole mobility of ~2.1 cm2 V-1 s-1 and EQE of 1.6% at a luminance of 2600 cd m-2. Most importantly, we observed a large area emission under the entire drain electrode, which was spatially stable (emission area is not dependent on the gate voltage and current density). These results show an important advancement in polymer-based LEFET technology toward realizing new digital display applications.

Biosynthesis, characterization and anti-dengue vector activity of silver nanoparticles prepared from Azadirachta indica and Citrullus colocynthis.

We report here biosynthesis of silver nanoparticles (AgNPs) using aqueous extracts of (i) Azadirachta indica leaves and (ii) Citrullus colocynthis fruit and their larvicidal activity against Aedes aegypti. The UV-Vis spectroscopy absorption peaks occurred in the range of 412-416 nm for A. indica AgNPs and 416-431 nm for C. colocynthis AgNPs indicating the silver nature of prepared colloidal samples. The scanning electron microscopy examination revealed the spherical morphology of both types of NPs with average size of 17 ± 4 nm (A. indica AgNPs) and 26 ± 5 nm (C. colocynthis AgNPs). The X-ray diffraction pattern confirmed the face-centred cubic (FCC) structure with crystallite size of 11 ± 1 nm (A. indica AgNPs) and 15 ± 1 nm (C. colocynthis AgNPs) while characteristic peaks appearing in Fourier transform infrared spectroscopy analysis indicated the attachment of different biomolecules on AgNPs. The larvicidal activity at different concentrations of synthesized AgNPs (1-20 mg l-1) and extracts (0.5-1.5%) against Aedes aegypti was examined for 24 h. A concentration-dependent larvicidal potential of both types of AgNPs was observed. The LC50 values were found to be 0.3 and 1.25 mg l-1 for C. colocynthis AgNPs and A. indica AgNPs, respectively. However, both extracts did not exhibit any notable larvicidal activity.