PLASMA DONATION AND PERCEPTION, ATTITUDE, BEHAVIOR OF COVID-19 PATIENTS: A CROSS SECTIONAL STUDY

BACKGROUND: Current study aims to identify the perception, attitude and behavior about Covid and plasma donation in the Covid cases. METHODOLOGY: This descriptive cross-sectional study was conducted ---removed for blind review---Adult COVID-19/post-Covid patients were included by consecutive sampling. The critically ill, mechanically ventilated cases were excluded. Special questionnaire was developed including the demographic variables, mode of transmission, personal hygiene, prevention, post covid immunity, re-infection, psychosocial factors, financial reservations and post covid life. Willingness for plasma donation, laboratory diagnostics and blood groups inquired. Data was collected by direct interview by researcher and analyzed by SPSS V.20. RESULTS: Mean age was 39.8+15 years; 122(54%) females and 104(46%) males. Total 163(73%) participants said Covid has impact on health, economy, social, mental and psychological state. 188(83%) considered Covid a threat to human life. 142(63%) had a close Covid contact and 15(6.6%) had recently travelled. 131(58%) said they could have prevented getting infected. 171(75.7%) considered handwashing and 208(92%) cleanliness and158(77%) considered natural, herbal remedies as preventive. 191(84.5%) wore mask for most/all of the time. 130(57.5%) said they will be immune to Covid post-recovery. 179(79.2%) were aware of re-infection. 169(74.8%) considered smoking as a risk for Covid and137(60.6%) aimed to quit smoking. 204(93%) committed to hand washing and 210(92.9%) to wearing masks post-Covid. 127(56%) were concerned about their food, 78(34.5%) about finances, 103(45.6%) about their family getting infected. 213(94%) expected life to normalize post-Covid. Most frequent blood group was B+ 67(29.6%) followed by A+ 42(18.6%) and O+ 41(18.1%). 128(66.6%) participants showed willingness to donate their plasma after recovery. 24(10.6%) refused the donation. 134(59.3%) agreed that plasma donation won’t reduce their immunity. 186(82.3%) were clinically recovered at the time of interview. CONCLUSION: Our Covid patients had a positive approach towards plasma donation. They expected normalization of life post Covid and showed commitment toward continuation of preventive habits and smoking cessation. However, there were significant concerns about finances, safety of loved ones and mental health.

Massive X-ray screening reveals two allosteric drug binding sites of SARS-CoV-2 main protease

The coronavirus disease (COVID-19) caused by SARS-CoV-2 is creating tremendous health problems and economical challenges for mankind. To date, no effective drug is available to directly treat the disease and prevent virus spreading. In a search for a drug against COVID-19, we have performed a massive X-ray crystallographic screen of repurposing drug libraries containing 5953 individual compounds against the SARS-CoV-2 main protease (Mpro), which is a potent drug target as it is essential for the virus replication. In contrast to commonly applied X-ray fragment screening experiments with molecules of low complexity, our screen tested already approved drugs and drugs in clinical trials. From the three-dimensional protein structures, we identified 37 compounds binding to Mpro. In subsequent cell-based viral reduction assays, one peptidomimetic and five non-peptidic compounds showed antiviral activity at non-toxic concentrations. Interestingly, two compounds bind outside the active site to the native dimer interface in close proximity to the S1 binding pocket. Another compound binds in a cleft between the catalytic and dimerization domain of Mpro. Neither binding site is related to the enzymatic active site and both represent attractive targets for drug development against SARS-CoV-2. This X-ray screening approach thus has the potential to help deliver an approved drug on an accelerated time-scale for this and future pandemics

X-ray screening identifies active site and allosteric inhibitors of SARS-CoV-2 main protease

The coronavirus disease (COVID-19) caused by SARS-CoV-2 is creating tremendous human suffering. To date, no effective drug is available to directly treat the disease. In a search for a drug against COVID-19, we have performed a high-throughput X-ray crystallographic screen of two repurposing drug libraries against the SARS-CoV-2 main protease (M^(pro)), which is essential for viral replication. In contrast to commonly applied X-ray fragment screening experiments with molecules of low complexity, our screen tested already approved drugs and drugs in clinical trials. From the three-dimensional protein structures, we identified 37 compounds that bind to M^(pro). In subsequent cell-based viral reduction assays, one peptidomimetic and six non-peptidic compounds showed antiviral activity at non-toxic concentrations. We identified two allosteric binding sites representing attractive targets for drug development against SARS-CoV-2

Effect of solvent polarity on extraction yield and antioxidant properties of phytochemicals from bean (Phaseolus vulgaris) seeds

The effect of solvent polarity on extraction yield and antioxidant properties of phytochemical compounds in bean seeds was studied. Seed flour of three varieties of bean was extracted in a series of organic solvents with increasing polarity (n-hexane, petroleum ether, chloroform, ethyl acetate, ethanol, acetone and water). Preliminary screening of phytochemicals showed the presence of tannins, flavonoids, cardiac glycosides, anthocyanins, terpenoids, carotenoids, ascorbic acid and reducing compounds in all extracts. One way analysis of variance (ANOVA) of results showed that extraction yield, phytochemical content and antioxidant properties were significantly influenced (p<0.05) by the polarity of extracting solvents. The regression analysis of data showed polarity-dependent second order polynomial variations in the extraction yield, phytochemical contents, antioxidant activity, reducing properties and free radical scavenging activity of each variety. Extraction in highly polar solvents resulted in high extract yield but low phenolic and flavonoid content as compared to non-polar ones. The polarity-dependent increase in total antioxidant activity and reducing properties indicates the extraction of strong antioxidant compounds in polar solvents. The study suggests the use of a combination of polar and nonpolar solvents to increase the extraction efficiency of phytochemicals with good antioxidant quality from the bean and other legume seeds

CONSERVATIVE ORAL THERAPY VERSUS RIFAXIMIN PLUS LACTOSE: WHICH IS A BETTER OPTION TO MANAGE RECURRENCE ONSET OF HE (HEPATIC ENCEPHALOPATHY)

Objective: To have a comparison of the recurrence or frequency of HP (hepatic encephalopathy) with a liver cirrhosis patient having treatment with Rifaximin plus Lactulose as opposed to oral conventional treatment by Lactulose only. Materials & Methods: This research started from January and ended in August 2017 at the Department of Medicine, Jinnah Hospital, Lahore. In this research, a total number of 200 HE patients were considered and divided into two groups: a group of getting treatment patients and a placebo group. The recurrence or frequency of HE was evaluated amid these two groups. Results: A comparison of 200 HE patients was carried out. The mean of the patient age was (44.78 ± 11.87) years in which the group under treatment was of mean ages (44.33 ± 10.45) years and the placebo group mean ages were (43.33 ± 10.13) years. After comparison, the cases of the group under treatment were 22 (22%) and cases of the placebo group were 47 (47%). There were significantly higher (P = 0.000) HE recurrence in the placebo group as opposed to the group under treatment. Conclusion: For decreasing the recurrence or frequency of HE the use of rifaximin plus lactulose is better than conventional treatment by lactulose only. Keywords: Rifaximin, Hepatic Encephalopathy (HE), Oral, Recurrence and Lactulose

High-resolution crystal structure and biochemical characterization of a GH11 endoxylanase from Nectria haematococca

Enzymatic degradation of vegetal biomass offers versatile procedures to improve the production of alternative fuels and other biomass-based products. Here we present the three-dimensional structure of a xylanase from Nectria haematococca (NhGH11) at 1.0 Å resolution and its functional properties. The atomic resolution structure provides details and insights about the complex hydrogen bonding network of the active site region and allowed a detailed comparison with homologous structures. Complementary biochemical studies showed that the xylanase can catalyze the hydrolysis of complex xylan into simple xylose aldopentose subunits of different lengths. NhGH11 can catalyze the efficient breakdown of beechwood xylan, xylan polysaccharide, and wheat arabinoxylan with turnover numbers of 1730.6 ± 318.1 min$^{−1}$, 1648.2 ± 249.3 min$^{−1}$ and 2410.8 ± 517.5 min$^{−1}$ respectively. NhGH11 showed maximum catalytic activity at pH 6.0 and 45 °C. The mesophilic character of NhGH11 can be explained by distinct structural features in comparison to thermophilic GH11 enzymes, including the number of hydrogen bonds, side chain interactions and number of buried water molecules. The enzymatic activity of NhGH11 is not very sensitive to metal ions and chemical reagents that are typically present in associated industrial production processes. The data we present highlights the potential of NhGH11 to be applied in industrial biomass degradation processes

Solution Structures and Dynamic Assembly of the 24-Meric Plasmodial Pdx1–Pdx2 Complex

Plasmodium species are protozoan parasites causing the deadly malaria disease. They have developed effective resistance mechanisms against most antimalarial medication, causing an urgent need to identify new antimalarial drug targets. Ideally, new drugs would be generated to specifically target the parasite with minimal or no toxicity to humans, requiring these drug targets to be distinctly different from the host’s metabolic processes or even absent in the host. In this context, the essential presence of vitamin B6 biosynthesis enzymes in Plasmodium, the pyridoxal phosphate (PLP) biosynthesis enzyme complex, and its absence in humans is recognized as a potential drug target. To characterize the PLP enzyme complex in terms of initial drug discovery investigations, we performed structural analysis of the Plasmodium vivax PLP synthase domain (Pdx1), glutaminase domain (Pdx2), and Pdx1–Pdx2 (Pdx) complex (PLP synthase complex) by utilizing complementary bioanalytical techniques, such as dynamic light scattering (DLS), X-ray solution scattering (SAXS), and electron microscopy (EM). Our investigations revealed a dodecameric Pdx1 and a monodispersed Pdx complex. Pdx2 was identified in monomeric and in different oligomeric states in solution. Interestingly, mixing oligomeric and polydisperse Pdx2 with dodecameric monodisperse Pdx1 resulted in a monodispersed Pdx complex. SAXS measurements revealed the low-resolution dodecameric structure of Pdx1, different oligomeric structures for Pdx2, and a ring-shaped dodecameric Pdx1 decorated with Pdx2, forming a heteromeric 24-meric Pdx complex

Solution Structures and Dynamic Assembly of the 24-Meric Plasmodial Pdx1–Pdx2 Complex

Plasmodium species are protozoan parasites causing the deadly malaria disease. They have developed effective resistance mechanisms against most antimalarial medication, causing an urgent need to identify new antimalarial drug targets. Ideally, new drugs would be generated to specifically target the parasite with minimal or no toxicity to humans, requiring these drug targets to be distinctly different from the host’s metabolic processes or even absent in the host. In this context, the essential presence of vitamin B$_6$ biosynthesis enzymes in Plasmodium, the pyridoxal phosphate (PLP) biosynthesis enzyme complex, and its absence in humans is recognized as a potential drug target. To characterize the PLP enzyme complex in terms of initial drug discovery investigations, we performed structural analysis of the Plasmodium vivax PLP synthase domain (Pdx1), glutaminase domain (Pdx2), and Pdx1–Pdx2 (Pdx) complex (PLP synthase complex) by utilizing complementary bioanalytical techniques, such as dynamic light scattering (DLS), X-ray solution scattering (SAXS), and electron microscopy (EM). Our investigations revealed a dodecameric Pdx1 and a monodispersed Pdx complex. Pdx2 was identified in monomeric and in different oligomeric states in solution. Interestingly, mixing oligomeric and polydisperse Pdx2 with dodecameric monodisperse Pdx1 resulted in a monodispersed Pdx complex. SAXS measurements revealed the low-resolution dodecameric structure of Pdx1, different oligomeric structures for Pdx2, and a ring-shaped dodecameric Pdx1 decorated with Pdx2, forming a heteromeric 24-meric Pdx complex

Rapid and efficient room temperature serial synchrotron crystallography using the CFEL TapeDrive

Serial crystallography at conventional synchrotron light sources (SSX) offers the possibility to routinely collect data at room temperature using micron sized crystals of biological macromolecules. However, it suffers from the fact that data collection is not yet as routine and takes currently significantly longer as the standard rotation series cryo-crystallography. Thus its use for high-throughput approaches, such as fragment-based drug screening, where the possibility to measure at physiological temperatures would be a great benefit, is impaired. On the way to high-throughput serial synchrotron crystallography, it is shown here, using a conveyor belt based sample delivery system – the CFEL TapeDrive – with three different proteins of biological relevance (K. pneumoniae CTX-M-14 β-lactamase, Nectria haematococca xylanase GH11 and Aspergillus flavus urate oxidase), that complete data sets can be collected in less than a minute and that only minimal amounts of sample are required