Virtually all SARS-CoV-2 vaccines currently in clinical testing are stored in a refrigerated or frozen state prior to use. This is a major impediment to deployment in resource-poor settings. Furthermore, several of them use viral vectors or mRNA. In contrast to protein subunit vaccines, there is limited manufacturing expertise for these nucleic acid-based modalities, especially in the developing world. Neutralizing antibodies, the clearest known correlate of protection against SARS-CoV-2, are primarily directed against the Receptor Binding Domain (RBD) of the viral spike protein, suggesting that a suitable RBD construct might serve as a more accessible vaccine ingredient. We describe a monomeric, glycan engineered RBD protein fragment that is expressed at a purified yield of 214 mg/L in unoptimized, mammalian cell culture and, in contrast to a stabilized spike ectodomain, is tolerant of exposure to temperatures as high as 100 °C when lyophilized, up to 70 °C in solution and stable for over four weeks at 37 °C. In prime:boost guinea pig immunizations, when formulated with the MF59-like adjuvant AddaVax™, the RBD derivative elicited neutralizing antibodies with an endpoint geometric mean titer of ~415 against replicative virus, comparing favourably with several vaccine formulations currently in the clinic. These features of high yield, extreme thermotolerance and satisfactory immunogenicity suggest that such RBD subunit vaccine formulations hold great promise to combat COVID-19.

The receptor binding domain (RBD) of SARS-CoV-2 is the primary target of neutralizing antibodies. We designed a trimeric, highly thermotolerant glycan engineered RBD by fusion to a heterologous, poorly immunogenic disulfide linked trimerization domain derived from cartilage matrix protein. The protein expressed at a yield of ∼80–100 mg/L in transiently transfected Expi293 cells, as well as CHO and HEK293 stable cell lines and formed homogeneous disulfide-linked trimers. When lyophilized, these possessed remarkable functional stability to transient thermal stress of up to 100 °C and were stable to long-term storage of over 4 weeks at 37 °C unlike an alternative RBD-trimer with a different trimerization domain. Two intramuscular immunizations with a human-compatible SWE adjuvanted formulation elicited antibodies with pseudoviral neutralizing titers in guinea pigs and mice that were 25–250 fold higher than corresponding values in human convalescent sera. Against the beta (B.1.351) variant of concern (VOC), pseudoviral neutralization titers for RBD trimer were ∼3-fold lower than against wildtype B.1 virus. RBD was also displayed on a designed ferritin-like Msdps2 nanoparticle. This showed decreased yield and immunogenicity relative to trimeric RBD. Replicative virus neutralization assays using mouse sera demonstrated that antibodies induced by the trimers neutralized all four VOC to date, namely B.1.1.7, B.1.351, P.1, and B.1.617.2 without significant differences. Trimeric RBD immunized hamsters were protected from viral challenge. The excellent immunogenicity, thermotolerance, and high yield of these immunogens suggest that they are a promising modality to combat COVID-19, including all SARS-CoV-2 VOC to date.

Saturation suppressor mutagenesis was used to generate thermostable mutants of the SARS-CoV-2 spike receptor-binding domain (RBD). A triple mutant with an increase in thermal melting temperature of ~7°C with respect to the wild-type B.1 RBD and was expressed in high yield in both mammalian cells and the microbial host, Pichia pastoris, was downselected for immunogenicity studies. An additional derivative with three additional mutations from the B.1.351 (beta) isolate was also introduced into this background. Lyophilized proteins were resistant to high-temperature exposure and could be stored for over a month at 37°C. In mice and hamsters, squalene-in-water emulsion (SWE) adjuvanted formulations of the B.1-stabilized RBD were considerably more immunogenic than RBD lacking the stabilizing mutations and elicited antibodies that neutralized all four current variants of concern with similar neutralization titers. However, sera from mice immunized with the stabilized B.1.351 derivative showed significantly decreased neutralization titers exclusively against the B.1.617.2 (delta) VOC. A cocktail comprising stabilized B.1 and B.1.351 RBDs elicited antibodies with qualitatively improved neutralization titers and breadth relative to those immunized solely with either immunogen. Immunized hamsters were protected from high-dose viral challenge. Such vaccine formulations can be rapidly and cheaply produced, lack extraneous tags or additional components, and can be stored at room temperature. They are a useful modality to combat COVID-19, especially in remote and low-resource settings.

Food safety is a rising challenge worldwide due to the expanding population and the need to produce food to feed the growing population. At the same time, pesticide residues found in high concentrations in fresh agriculture pose a significant threat to food safety. Presently, crop output is being increased by applying herbicides, fungicides, insecticides, pesticides, fertilizers, nematicides, and soil amendments. A combination of factors, including bioaccumulation, widespread usage, selective toxicity, and stability, make pesticides among the most toxic compounds polluting the environment. They are especially harmful in vegetables and fruits because people are exposed to them. Thus, it is critical to monitor pesticide levels in fruits and vegetables using all analytical techniques available. Any evaluation of the condition of pesticide contamination in fruits and vegetables necessitates knowledge of maximum residue levels (MRLs). We set out the problems in determining various types of pesticides in vegetables and fruits, including the complexity and the diversity of matrices in biological materials. This review examines the different analytical techniques to determine the target analytes that must be isolated before final consumption. Many processes involved determining pesticide residues in fruits and vegetables and their advantages and disadvantages have been discussed with recommendations. Furthermore, MRLs of target pesticide residues in fruit and vegetable samples are discussed in the context of data from the literature. The review also examines MRLs’ impact on the international trade of fruits and vegetables. Accurate, sensitive, and robust analytical procedures are critical to ensuring that pesticide levels in food products are effectively regulated. Despite advances in detection technology, effective sample preparation procedures for pesticide residue measurement in cereals and feedstuffs are still needed. In addition, these methods must be compatible with current analytical techniques. Multi-residue approaches that cover a wide range of pesticides are desired, even though pesticides’ diverse natures, classes, and physio-chemical characteristics make such methods challenging to assemble. This review will be valuable to food analysts and regulatory authorities to monitor the quality and safety of fresh food products.

As existing vaccines fail to completely prevent COVID-19 infections or community transmission, there is an unmet need for vaccines that can better combat SARS-CoV-2 variants of concern (VOC). We previously developed highly thermo-tolerant monomeric and trimeric receptor-binding domain derivatives that can withstand 100 °C for 90 min and 37 °C for four weeks and help eliminate cold-chain requirements. We show that mice immunised with these vaccine formulations elicit high titres of antibodies that neutralise SARS-CoV-2 variants VIC31 (with Spike: D614G mutation), Delta and Omicron (BA.1.1) VOC. Compared to VIC31, there was an average 14.4-fold reduction in neutralisation against BA.1.1 for the three monomeric antigen-adjuvant combinations and a 16.5-fold reduction for the three trimeric antigen-adjuvant combinations; the corresponding values against Delta were 2.5 and 3.0. Our findings suggest that monomeric formulations are suitable for upcoming Phase I human clinical trials and that there is potential for increasing the efficacy with vaccine matching to improve the responses against emerging variants. These findings are consistent with in silico modelling and AlphaFold predictions, which show that, while oligomeric presentation can be generally beneficial, it can make important epitopes inaccessible and also carries the risk of eliciting unwanted antibodies against the oligomerisation domain.

Ninety two sugarcane varieties from sub-tropical India were subjected to molecular profiling with 174 simple sequence repeat markers and characterized for 23 qualitative (morphological descriptors) and nine quantitative traits that directly or indirectly contribute to yield and juice quality. Using STRUCTURE-based population stratification study and a mixed linear model for marker-trait association (MTA) analysis, a total of 60 MTAs were identified for 22 qualitative traits that were able to explain a significantly higher (up to 40%) proportion of the phenotypic variations compared to all the previous reports of MTA studies in sugarcane. In addition, 21 MTAs stable over the three years of study were also identified for nine quantitative traits that explained 16-37% of the total trait variation. It could be concluded that the qualitative traits that are governed mostly by one or a few genes are more responsive to MTA studies and hence have a better potential to be adopted in marker-assisted breeding programmes in sugarcane. The MTAs identified in this study could also find significant applications in upcoming more stringent IP regime, which may necessitate tracking of specific alleles integrated in breeding programmes.

The COVID-19 pandemic at its peak compelled students to stay home and adapt to the distance learning system. The world has gone through phases of fear and respite in the recent years. There have been a number of studies related to student learning via online teaching during the pandemic. Now, as the vaccination coverage picks up and the pandemic appears to have achieved a plateau, it is time to take a view of students’ perceptions of online learning and its effectiveness in skill development during the COVID-19 pandemic. This study assesses the students’ preferences and perceptions regarding offline and online learning post-COVID-19 lockdown and with the resumption of offline classes. A cross-sectional study was conducted at King Khalid University, Aseer region, from the period of 1 January 2022 to 30 January 2022. A convenience sampling technique was utilized to collect data from female students. Data analysis was conducted by using SPSS version 22.0. A total of 480 students participated in the study, their mean age was 19.79 ± 1.48. More than half of the students (64%) still fear getting COVID-19 as they continue with the in-person or offline classes despite having completed their doses of vaccination. Almost half of the students have difficulty in waking up after the recommencement of offline classes. The majority (77%) felt tired after starting offline classes, and 63% felt unhappy after starting offline classes again. The majority of students believe that, with offline classes, they have difficulty in time management and concentration. The majority of students believed that, with online classes, they are more comfortable in gaining knowledge and learning, more alert, more satisfied, and gain higher scores in exams. The majority of students preferred the online mode of learning, with about 72% of students wishing to continue online learning in the future. This research underlines the influence of fear of getting COVID-19 after the commencement of face-to-face learning amongst the students. Students were more inclined to continue with online classes because of fear of getting COVID-19, despite having full doses of the vaccination in Saudi Arabia. There is a need for a better understanding of students’ motivations and coping mechanisms during the pandemic.

Virtually all SARS-CoV-2 vaccines currently in clinical testing are stored in a refrigerated or frozen state prior to use. This is a major impediment to deployment in resource-poor settings. Several use viral vectors or mRNA. In contrast to protein subunit vaccines, there is limited manufacturing expertise for these novel, nucleic acid based modalities, especially in the developing world. Neutralizing antibodies, the clearest known correlate of protection against SARS-CoV-2, are primarily directed against the Receptor Binding Domain (RBD) of the viral spike protein. We describe a monomeric, glycan engineered RBD protein fragment that is expressed at a purified yield of 200mg/L in unoptimized, mammalian cell culture and in contrast to a stabilized spike ectodomain, is tolerant of exposure to temperatures as high as 100°C when lyophilized, and upto 70°C in solution. In prime:boost guinea pig immunizations, when formulated with the MF59 like adjuvant AddaVax™, the RBD derivative elicited neutralizing antibodies with an endpoint geometric mean titer of ~415 against replicative virus, comparing favourably with several vaccine formulations currently in the clinic. These features of high yield, extreme thermotolerance and satisfactory immunogenicity suggest that such RBD subunit vaccine formulations hold great promise to combat COVID-19.