Prediction of putative epitope-based vaccine against all corona virus strains for the Chinese population: Approach toward development of vaccine.

Currently, the whole world is facing the coronavirus disease-19 pandemic. As of now, approximately 0.15 million people around the globe are infected with the novel coronavirus. In the last decade, two strains of the coronavirus family, severe acute respiratory syndrome-related coronavirus and Middle East respiratory syndrome coronavirus, also resulted in epidemics in south Asian and the Middle Eastern countries with high mortality rate. This scenario demands the development of a putative vaccine which may provide immunity against all current and new evolving coronavirus strains. In this study, we designed an epitope-based vaccine using an immunoinformatic approach. This vaccine may protect against all coronavirus strains. The vaccine is developed by considering the geographical distribution of coronavirus strains and host genetics (Chinese population). Nine experimentally validated epitopes sequences from coronavirus strains were used to derive the variants considering the conservancy in all strains. Further, the binding affinities of all derived variants were checked with most abundant human leukocyte antigen alleles in the Chinese population. Three major histocompatibility complex (MHC) Class I epitopes from spike glycoprotein and nucleoprotein showed sufficient binding while one MHC Class II epitope from spike glycoprotein was found to be an effective binder. A cocktail of these epitopes gave more than 95% population coverage in the Chinese population. Moreover, molecular dynamics simulation supported the aforementioned predictions. Further, in vivo studies are needed to confirm the immunogenic potential of these vaccines.

Association between bacterial strain type and host biomarkers in Clostridium perfringens infected goats.

The study project was designed to determine the effects of Clostridium perfringens type D infection on hematological and biochemical parameters in goats. Purposive blood samples were collected from 6 healthy and 12 diseased goats positive for C. perfringens infection. Neither the animals nor their mother were vaccinated against Clostridium perfringens from whom samples were obtained. Study was carried out in two different topographic areas; hilly (district Swat) and plain (district Mardan) of Khyber Pakhtunkhwa, Pakistan but nonsignificant (P > 0.05) statistical difference was recorded between the prevalence of Clostridium perfringens infected goats. Mean erythrocytes count (RBC) and hemoglobin level decreased significantly (P < 0.05) while the white blood cells (WBC) increased significantly (P < 0.05) in diseased animals compared to the healthy animals. However non-significant differences (P > 0.05) were observed in packet cell volume (PCV) and platelets count in healthy and diseased animals. According to biochemical analysis, a significant increase (P < 0.05) in liver enzymes, total bilirubin, serum creatinine, blood urea and glucose was recorded in diseased goats. . The results demonstrated that fluctuation in most of the mean hematological values remained within the normal range however the mean liver enzymes, total bilirubin, serum creatinine, blood urea and glucose levels gone beyond the normal levels which demonstrated severe damages to liver and kidneys.

Comparative Analysis of Nutritional Properties, Phytochemical Profile, and Antioxidant Activities between Red and Green Water Chestnut (Trapa natans) Fruits.

The present study explored the nutritional composition, phytochemicals analysis, and antioxidant capacity of two indigenous varieties of red and green water chestnut (WCN) fruit grown in Pakistan. Accordingly, this study was designed to investigate the proximate composition (moisture, ash, fiber, proteins, fat, and energy), physicochemical properties (pH, °Brix, and glycemic index), minerals, and vitamins. The methanolic extracts of WCN fruits were explored for phytochemicals (total phenolic and flavonoid content), and antioxidant potential was examined in vitro by 1,1-diphenyl-2-picrylhydrazyl radical scavenging capacity (DPPH) and Ferric reducing antioxidant power (FRAP). Quantitative determination of mineral (sodium, potassium, calcium, phosphorus, iron, manganese, copper, and zinc) and vitamin (vitamin C, vitamin B6, vitamin B2, vitamin B3, vitamin A, and ß-Carotene) composition was also assessed. Based on the findings, the proximate compositions of WCN green and red varieties varied greatly as WCN green contained significantly higher protein (1.72%), fat (0.65%), dietary fiber (2.21%), moisture (70.23%), ash (1.16%), and energy content (112.8 Kcal) than WCN red. In WCN green, the macro-micromineral concentrations were significantly higher than WCN red. Among the minerals analyzed, potassium was the most abundant mineral found in both varieties. Levels of vitamin C, B6, A, and ß-Carotene were significantly higher in WCN green. In this study, methanolic extract showed higher extraction efficiency than acetone, ethanol, and distilled water. WCN green had a significantly higher quantum of total phenolic (91.13 mg GAE/g) and total flavonoid (36.6 mg QE/g) and presented significantly higher antioxidant activity than the WCN red. This study showed that, among both varieties, WCN green extract has therapeutic potential against free radical mediated health conditions and suggested the potential use of this fruit as a source of natural antioxidants in nutraceuticals.

Antidiabetic, Antioxidative and Antihyperlipidemic Effects of Strawberry Fruit Extract in Alloxan-Induced Diabetic Rats.

Strawberry (Fragaria × ananassa) is one of the accomplished sources of bioactive compounds, including anthocyanin, phenolic acids, flavonols, ellagitannins, and a diverse range of minerals and vitamins that can help to boost human health. This study was carried out to explore the antidiabetic, antioxidative and antihyperlipidemic potential of strawberry extracts against alloxan-induced (100 mg/kg body weight) diabetic rats. Accordingly, rats were categorized into six groups including control (G0), positive control (G1), treatment groups (G2, G3, and G4) given strawberry extract at 250, 500, and 750 mg/kg of body weight, respectively, and G5 provided metformin @70 mg/kg BW for 28 days with ad libitum diet. At the trial termination, the rats were sacrificed and were subjected to analysis including body weight, blood glucose level and glycemic indicators, antioxidant parameters, lipid profile, renal function test (RFT), liver function test (LFT) and histopathology for pancreatic tissues. The results indicated that treatment of diabetic rats with strawberry extract at 500 mg/kg body weight (BW) resulted in significant reductions in blood glucose level, serum urea, and creatinine as well as significant increases in body weight, insulin activity, and protein levels. In addition, the diabetic rats that did not receive strawberry extract (control) exhibited an increase in plasma glucose, urea, uric acid, creatinine, and a decrease in body weight and insulin levels. Briefly, it is reported that strawberry fruit extracts reduced blood sugar levels, possess hypolipidemic potential, and helped to maintain antioxidant levels in alloxan-induced diabetic rats.

Determination of Pleiotropic Effect of Warfarin in VKORC1 and CYP2C9 Genotypes in Patients With Heart Valve Replacement.

Warfarin has been widely used as an oral anticoagulant agent. In past, efforts have been done to study the contribution of genetic variation on warfarin dose requirements. The possible therapeutic dose determination of warfarin is very challenging, i.e., extremely low dose leading to unusable antithrombotic therapy or high dose causes particularly bleeding complications. Our study aimed to investigate these observations in more detail, we determined the correlation of interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), and tumor necrosis factor-α (TNF-α) among VKORC1 and CYP2C9 genetic variants in patients with heart valve replacement who were treated with a range of warfarin doses and compared with levels in healthy controls. A total of 107 human subjects were recruited with low < 5 mg, medium 5-10 mg/day, and high > 10 mg/day warfarin doses. The genetic study of VKORC1-1639G/A, C1173T, 3730G > A, CYP2C9*2, and CYP2C9*3 was performed using TaqMan genotyping and DNA sequencing. The gene expression of IL-6, TNF-α, and COX-2 mRNA was analyzed. IL-6, TNF-α, and COX-2 protein expressions were determined by ELISA and Western blot analysis to evaluate the pro- and anti-inflammatory effects of warfarin. A statistically significant difference was found among the haplotypes of VKORC1 rs9934438 (C1173T), rs9923231 (-1639G > A), rs7294 (3730G > A) and CYP2C9 *2 p. Arg144 Cys (rs28371674), CYP2C9 *3 p. Ile359Leu (rs1057910) genotypes with warfarin dose requirements (p = 0.001). The increased levels of COX-2, IL-6, and TNF-α proteins were observed when a high dose of warfarin (>10 mg/ml) was administered. However, a lower concentration (1.0 mg/ml) was observed with decreased warfarin dose (<5 mg/day). The present study reported that in addition to its anticoagulant action, the genetic variants of warfarin may have a pleiotropic effect by influencing IL-6 depending on the dosing regimen and inducing the expression of COX-2.

Corrigendum: Disease specific symptoms indices in patients with celiac disease-A hardly recognised entity.

[This corrects the article DOI: 10.3389/fnut.2022.944449.].

Disease specific symptoms indices in patients with celiac disease-A hardly recognised entity.

Background: Celiac disease (CD) was considered a rare disease before and was perceivably only limited to children but now affects almost 1-2% of the global population. This abrupt increase in prevalence is due to advancements in diagnostic criteria and medical facilities but still many countries lack the basic data that can assess the severity of this health issue. The present study was conducted with the aim to assess the common but rarely diagnosed condition with the identification of its underlying secondary ailments. Materials and methods: Patients visiting public sector hospitals were recruited and tested for clinical symptoms secondary to gluten-containing foods (wheat and barley, etc.), followed by serological testing for immunoglobulin A, tissue transglutaminase A, and anti-endomysial antibodies. Only seropositive candidates were included in the endoscopic and biopsy examination for the features of villous atrophy and intestinal cell damage. The secondary ailments including anemia, growth retardation, and gastrointestinal symptoms were also documented for the tested positive patients. The modified European Society of Pediatric Gastroenterology, Hepatology, and Nutrition (ESPGHAN) criterion was followed throughout the study. Results: From 647 suspected cases from March 2018 to July 2019, 113 were confirmed with CD while 58% were female children and 42% were male children. The majority of them were from a lower class (75%) and 26% of them had a positive family history of CD. A total of 67% of patients with CD were underweight while wasting was observed in 38%, and 80% were stunted as well. Of the positively tested patients with CD, 49% had moderate anemia with 15% having severe anemia. Approximately 33% had hypoalbuminemia as well. The majority of them had a mild to severe range of gastrointestinal symptoms, such as abdominal pain, diarrhea, flatus, eructation, diarrhea, and steatorrhea. Conclusion: The study finding indicates an increased number of patients diagnosed with CD with an excessive sum of secondary ailments, such as anemia, growth failure, growth retardation, malnutrition, and gastrointestinal symptoms.

Evaluation of COVID-19 Disease Awareness and Its Relation to Mental Health, Dietary Habits, and Physical Activity: A Cross-Sectional Study from Pakistan.

Coronavirus infection (COVID-19) has significantly increased the mortality and morbidity rates worldwide. The present study was conducted to assess the general public's awareness of COVID-19 and its association with mental health, dietary habits, and physical activity. A web-based survey was conducted to gather information about demographics, knowledge about COVID-19, dietary habits, mental health, and anthropometry among the general public of Pakistan. Descriptive statistics, chi-square test, and multiple logistic regression were used for data analysis. The majority of the participants were suffering from anxiety (71.0%) and depression (52.0%) during the COVID-19 pandemic; 32.4% of participants had poor COVID-19-related knowledge. COVID-19 lockdown reduced the physical activity of 66.9% of participants and increased weight of 38.8% of the survey participants. Demographic variables, including age, gender, ethnicity, education, employment, family type, and geographical location, were significantly associated with knowledge about COVID-19 (P < 0.05). Depression was inversely associated with COVID-19 knowledge (P < 0.05). Healthy changes in dietary habits including decreased consumption of fast foods, soft and cola drinks, fruit drinks, cooked meat (outside the home), sugar, and fats, and were associated with increased knowledge of COVID-19. Vitamin C and immunity-boosting supplement consumption were significantly associated with increased knowledge regarding COVID-19 (< 0.05). Inadequate knowledge about COVID-19 and the presence of anxiety and depression were found among most of the study participants. There is a need to conduct educational seminars to limit the health consequences resulting from COVID-19 lockdown.

Development of multi-epitope vaccine constructs for non-small cell lung cancer (NSCLC) against USA human leukocyte antigen background: an immunoinformatic approach toward future vaccine designing.

OBJECTIVES: The design of peptide-based vaccines for cancer is a promising immunotherapy that can induce a cancer-specific cytotoxic response in tumor cells. METHODS: Herein, we used the immunoinformatic approach in designing a multi-epitope vaccine targeting G-protein coupled receptor 87 (GPCR-87), cystine/glutamate transporter (SLC7A11), Immunoglobulin binding protein 1 (IGBP1), and thioredoxin domain-containing protein 5 (TXNDC5), which can potentially contribute to NSCLC. The MHC-I and MHC-II epitopes selected for the fusion construct were evaluated for their antigenic and non-allergenic natures via VaxiJen and AllerTop. RESULTS: A total of five epitopes, four class-I (FIFYLKNIV, CRYTSVLFY, RYLKVVKPF, and RQAKIQRYK), and one class-II (NQVRGYPTLLWFRDG), having combined USA population coverage of 100%, were used to make ten possible multi-epitope fusion constructs. In these constructs, PADRE, a universal T-helper epitope, and RSO9, a TLR4 agonist, were fused as adjuvants. The molecular docking analysis revealed that two constructs were showing significant binding affinities toward HLA-A*02:01, the most prevalent HLA allele in USA. Moreover, MD simulations marked one construct as a promising therapeutic candidate. CONCLUSION: The multi-epitope vaccine constructs designed using immunogenic, and non-allergenic peptides of NSCLS tumor-associated proteins are likely to pose significant therapeutic efficacies in cancer immunotherapy due to their high binding affinities toward HLA molecules.

Survival and stability of free and encapsulated probiotic bacteria under simulated gastrointestinal conditions and in ice cream.

The aim of the present study was to evaluate the upshot of microencapsulation on the stability and viability of probiotics in carrier food (ice cream) and simulated gastrointestinal (GIT) conditions. Purposely, Lactobacillus casei was encapsulated with two different hydrocolloids, that is, calcium alginate (Ca-ALG) and whey protein concentrate (WPC) by using encapsulator. The obtained microbeads were characterized in terms of encapsulation efficiency and morphological features. Afterward, the probiotics in free and encapsulated form were incorporated into ice cream. The product was subjected for physicochemical, microbiological, and sensory attributes over a storage period of 80 days. Microencapsulation with both hydrogels significantly (p < .05) improved the viability of probiotics in both carrier food and simulated GIT conditions.The initial viable count of probiotics encapsulated with Ca-ALG and WPC was 9.54 and 9.52 log CFU/ml, respectively, that declined to 8.59 and 8.39 log CFU/ml, respectively, over period of 80 days of storage. While nonencapsulated/free cells declined from 9.44 to 6.41 log CFU/ml during same storage period. Likewise, during in vitro GIT assay, encapsulated probiotic with Ca-ALG and WPC showed 0.95 and 1.13 log reduction, respectively. On other hand, free probiotics showed significant 3.03 log reduction. Overall, microencapsulated probiotic exhibited better survival as compared to free cells. Moreover, the amalgamation of encapsulated and free probiotics affected the physicochemical (decrease in pH and increase in viscosity) was and sensory parameters of ice cream during storage.

Functional exploration of free and encapsulated probiotic bacteria in yogurt and simulated gastrointestinal conditions.

The core objective of the current study was to evaluate the effect of microencapsulation on the viability and stability of probiotic bacteria in yogurt and simulated gastrointestinal conditions. For this purpose, probiotic bacteria were encapsulated with sodium alginate and carrageenan by encapsulator. Yogurt was prepared with the incorporation of free and encapsulated probiotic bacteria and was analyzed for physicochemical, microbiological, and sensorial attributes. Encapsulation and storage exhibited a significant (p < .05) effect on different parameters of yogurt. An increasing trend in syneresis and acidity while a decreasing trend in viscosity, pH, viability, and stability were observed. The value of syneresis increased from 2.27 ± 0.17 to 2.9 ± 0.14 and acidity from 0.48 ± 0.04 to 0.64 ± 0.01 during 4 weeks of storage. The value of viscosity decreased from 3.68 ± 0.21 to 2.42 ± 0.09 and pH from 4.88 ± 0.31to 4.43 ± 0.36 during 28 days of storage. Unencapsulated (free) cells exhibited poor survival. The viable cell count of probiotic bacteria in the free-state in yogurt was 9.97 logs CFU/ml at zero-day that decreased to 6.12 log CFU/ml after 28 days. However, encapsulation improved the viability of the probiotics in the prepared yogurt and GIT. The cell count of probiotics encapsulated with sodium alginate and carrageenan was 9.91 logs CFU/ml and 9.89 logs CFU/ml, respectively, at zero-day that decreased to 8.74 logs CFU/ml and 8.39 log CFU/ml, respectively. Free cells (unencapsulated) showed very poor survival. Similarly, during in vitro gastrointestinal assay, the survival rate of encapsulated probiotic bacteria in simulated gastric solution and intestinal solutions was higher than that of free cells. In the case of encapsulated bacteria, only 3 logs while for free cells, 7 log reduction was recorded. Sodium alginate microcapsules exhibited better release profile than carrageenan. Conclusively, microencapsulation improved the survival of probiotic bacteria in carrier food as well as in simulated gastrointestinal condition.