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Abstract Hemagglutinin (HA)-based current vaccines provide suboptimum cross protection. Influenza A virus contains an ion channel protein M2 conserved extracellular domain (M2e), a target for developing universal vaccines. Here we generated reassortant influenza virus rgH3N2 4xM2e virus (HA and NA from A/Switzerland/9715293/2013/(H3N2)) expressing chimeric 4xM2e-HA fusion proteins with 4xM2e epitopes inserted into the H3 HA N-terminus. Recombinant rgH3N2 4xM2e virus was found to retain equivalent growth kinetics as rgH3N2 in egg substrates. Intranasal single inoculation of mice with live rgH3N2 4xM2e virus was effective in priming the induction of M2e specific IgG antibody responses in mucosal and systemic sites as well as T cell responses. The rgH3N2 4xM2e primed mice were protected against a broad range of different influenza A virus subtypes including H1N1, H3N2, H5N1, H7N9, and H9N2. The findings support a new approach to improve the efficacy of current vaccine platforms by recombinant influenza virus inducing immunity to HA and cross protective M2e antigens.

Knowledge of the physical and thermal properties of egg white powder (EWP) is needed for bulk handling and heat transfer modeling. The densities (bulk, tap, and true) of EWP at four different moisture content levels were determined. The densities of EWP significantly decreased with an increase in moisture content. The determined Hausner ratio classified the EWP as very poor flowing powder. The specific heat capacity (cp) of EWP was determined as a function of moisture content and temperature, and a first-order model with an interaction term described the relationship with a coefficient of determination (R²) of 0.964. Both moisture content and temperature significantly increased the cp of EWP. The thermal conductivity (k) of EWP as a function of moisture content and temperature was measured at three bulk density levels, and a linear regression model described the relationship with R² of 0.967. The bulk density, moisture content, and temperature of EWP significantly increased the k. The thermal diffusivity (α) of EWP was calculated from the experimental k and cp values at three different densities. The α of EWP significantly decreased with an increase in density and temperature; however, the decrease in α with an increase in moisture content was not significant. The models developed in this study are useful for predicting the thermal properties of EWP in simulations involving steady and transient heat transfer situations. [ABSTRACT FROM AUTHOR]

Alkaptonuria is a rare metabolic, autosomal recessive disorder caused by the deficiency of homogentisic acid oxidase and it is characterized by bluish-black discoloration of cartilages, skin (Ochronosis), degenerative changes in the articular, extra-articular cartilages, intervertebral disc, other tissues causing pain in the joints and spinal column. Although intervertebral disc degeneration is common in these patients, those presenting with symptoms severe enough to warrant surgery are rare. Only a few patients have been treated surgically. We present a case of alkaptonuria presenting with radiculopathy and lumbar disc herniation. The case presented demonstrates that although lumbar disc herniation is rare in alkaptonuria, it should be sought in such patients and surgical treatment yields good functional outcome.

With the success of COVID-19 vaccines, newly created mRNA vaccines against other infectious diseases are beginning to emerge. Here, we review the structural elements required for designing mRNA vaccine constructs for effective in vitro synthetic transcription reactions. The unprecedently speedy development of mRNA vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was enabled with previous innovations in nucleoside modifications during in vitro transcription and lipid nanoparticle delivery materials of mRNA. Recent updates are briefly described in the status of mRNA vaccines against SARS-CoV-2, influenza virus, and other viral pathogens. Unique features of mRNA vaccine platforms and future perspectives are discussed.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) gave rise to the coronavirus disease 2019 (COVID-19) pandemic. A strong correlation has been demonstrated between worse COVID-19 outcomes, aging, and metabolic syndrome (MetS), which is primarily derived from obesity-induced systemic chronic low-grade inflammation with numerous complications, including type 2 diabetes mellitus (T2DM). The majority of COVID-19 deaths occurs in people over the age of 65. Individuals with MetS are inclined to manifest adverse disease consequences and mortality from COVID-19. In this review, we examine the prevalence and molecular mechanisms underlying enhanced risk of COVID-19 in elderly people and individuals with MetS. Subsequently, we discuss current progresses in treating COVID-19, including the development of new COVID-19 vaccines and antivirals, towards goals to elaborate prophylactic and therapeutic treatment options in this vulnerable population.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to be expanding the pandemic disease across the globe. Although SARS-CoV-2 vaccines were rapidly developed and approved for emergency use of vaccination in humans, supply and production difficulties are slowing down the global vaccination program. The efficacy of many different versions of vaccine candidates and adjuvant effects remain unknown, particularly in the elderly. In this study, we compared the immunogenic properties of SARS-CoV-2 full-length spike (S) ectodomain in young adult and aged mice, S1 with receptor binding domain, and S2 with fusion domain. Full-length S was more immunogenic and effective in inducing IgG antibodies after low dose vaccination, compared to the S1 subunit. Old-aged mice induced SARS-CoV-2 spike-specific IgG antibodies with neutralizing activity after high dose S vaccination. With an increased vaccine dose, S1 was highly effective in inducing neutralizing and receptor-binding inhibiting antibodies, although both S1 and S2 subunit domain vaccines were similarly immunogenic. Adjuvant effects were significant for effective induction of IgG1 and IgG2a isotypes, neutralizing and receptor-binding inhibiting antibodies, and antibody-secreting B cell and interferon-γ secreting T cell immune responses. Results of this study provide information in designing SARS-CoV-2 spike vaccine antigens and effective vaccination in the elderly.

Respiratory syncytial virus (RSV) is one of the most important pathogens causing significant morbidity and mortality in infants and the elderly. Live attenuated influenza vaccine (LAIV) is a licensed vaccine platform in humans and it is known to induce broader immune responses. RSV G attachment proteins mediate virus binding to the target cells and they contain a conserved central domain with neutralizing epitopes. Here, we generated recombinant LAIV based on the attenuated A/Puerto Rico/8/1934 virus backbone, expressing an RSV conserved G-domain in a chimeric hemagglutinin (HA) fusion molecule (HA-G). The attenuated phenotypes of chimeric HA-G LAIV were evident by restricted replication in the upper respiratory tract and low temperature growth characteristics. The immunization of mice with chimeric HA-G LAIV induced significant increases in G-protein specific IgG2a (T helper type 1) and IgG antibody-secreting cell responses in lung, bronchioalveolar fluid, bone marrow, and spleens after RSV challenge. Vaccine-enhanced disease that is typically caused by inactivated-RSV vaccination was not observed in chimeric HA-G LAIV as analyzed by lung histopathology. These results in this study suggest a new approach of developing an RSV vaccine candidate while using recombinant LAIV, potentially conferring protection against influenza virus and RSV.

Crimean Congo hemorrhagic fever, a zoonotic viral disease, has high mortality rate in humans. There is currently no vaccine for Crimean Congo hemorrhagic fever virus (CCHFV) and chemical interventions are limited. The three negative sense genomic RNA segments of CCHFV are specifically encapsidated by the nucleocapsid protein into three ribonucleocapsids, which serve as templates for the viral RNA dependent RNA polymerase. Here we demonstrate that CCHFV nucleocapsid protein has two distinct binding modes for double and single strand RNA. In the double strand RNA binding mode, the nucleocapsid protein preferentially binds to the vRNA panhandle formed by the base pairing of complementary nucleotides at the 5' and 3' termini of viral genome. The CCHFV nucleocapsid protein does not have RNA helix unwinding activity and hence does not melt the duplex vRNA panhandle after binding. In the single strand RNA binding mode, the nucleocapsid protein does not discriminate between viral and non-viral RNA molecules. Binding of both vRNA panhandle and single strand RNA induce a conformational change in the nucleocapsid protein. Nucleocapsid protein remains in a unique conformational state due to simultaneously binding of structurally distinct vRNA panhandle and single strand RNA substrates. Although the role of dual RNA binding modes in the virus replication cycle is unknown, their involvement in the packaging of viral genome and regulation of CCHFV replication in conjunction with RdRp and host derived RNA regulators is highly likely.

Heterologous prime boost with DNA and recombinant modified vaccinia virus Ankara (rMVA) vaccines is considered as a promising vaccination approach against human immunodeficiency virus (HIV-1). To further enhance the efficacy of DNA-rMVA vaccination, we investigated humoral and cellular immune responses in mice after three sequential immunizations with DNA, a combination of DNA and virus-like particles (VLP), and rMVA expressing HIV-1 89.6 gp120 envelope proteins (Env). DNA prime and boost with a combination of VLP and DNA vaccines followed by an rMVA boost induced over a 100-fold increase in Env-specific IgG antibody titers compared to three sequential immunizations with DNA and rMVA. Cellular immune responses were induced by VLP-DNA and rMVA vaccinations at high levels in CD8 T cells, CD4 T cells, and peripheral blood mononuclear cells secreting interferon (IFN)-γ, and spleen cells producing interleukin (IL)-2, 4, 5 cytokines. This study suggests that a DNA and VLP combination vaccine with MVA is a promising strategy in enhancing the efficacy of DNA-rMVA vaccination against HIV-1.

Post-processing contamination of Listeria monocytogenes has remained a major concern for the safety of ready-to-eat (RTE) meat products that are not reheated before consumption. Mathematical models are rapid and cost-effective tools to predict pathogen behavior, product shelf life, and safety. The objective of this study was to develop and validate a comprehensive model to predict the Listeria growth rate in RTE meat products as a function of temperature, pH, water activity, nitrite, acetic, lactic, and propionic acids. The Listeria growth data in RTE food matrices, including RTE beef, pork, and poultry products (731 data sets), were collected from the literature and databases like ComBase. The growth parameters were estimated using the logistic-with-delay primary model. The good-quality growth rate data ( n = 596, R 2 > 0.9) were randomly divided into 80% training ( n = 480) and 20% testing ( n = 116) datasets. The training growth rates were used to develop a secondary gamma model, followed by validation in testing data. The growth model's performance was evaluated by comparing the predicted and observed growth rates. The goodness-of-fit parameter of the secondary model includes R 2 of 0.86 and RMSE of 0.06 (μ max ) during the development stage. During validation, the gamma model with interaction included an RMSE of 0.074 (μ max ), bias, and accuracy factor of 0.95 and 1.50, respectively. Overall, about 81.03% of the relative errors (RE) of the model's predictions were within the acceptable simulation zone (RE ± 0.5 log CFU/h). In lag time model validation, predictions were 7% fail-dangerously biased, and the accuracy factor of 2.23 indicated that the lag time prediction is challenging. The model may be used to quantify the Listeria growth in naturally contaminated RTE meats. This model may be helpful in formulations, shelf-life assessment, and decision-making for the safety of RTE meat products.

Millets are recognized as future foods due to their abundant nutrition and resilience, increasing their value on the global stage. Millets possess a broad spectrum of nutrients, antinutrients, and antioxidants, making it imperative to understand the effects of various processing methods on these components. Antinutritional factors interfere with the digestibility of macro-nutrients and the bioavailability and bio accessibility of minerals. This necessitates methods to reduce or eliminate antinutrients while improving nutritive and antioxidant value in food. This review aims to elucidate the rationale behind processing choices by evaluating the scientific literature and examining the mechanisms of processing methods, categorized as physiochemical, bio, thermal, novel non-thermal, and their combination techniques. Physiochemical and bioprocessing methods alter antinutrients and antioxidant profiles through mass transfer, enzyme activation, product synthesis, microbial activity, and selective removal of grain layers. Thermal methods break functional bonds, modify the chemical or physical structures, enhance kinetics, or degrade heat-labile components. Non-thermal techniques preserve heat-sensitive antioxidants while reducing antinutrients through structural modifications, oxidation by ROS, and break down the covalent and non-covalent bonds, resulting in degradation of compounds. To maximize the trade-off between retention of beneficial components and reducing detrimental ones, exploring the synergy of combination techniques is crucial. Beyond mitigating antinutrients, these processing methods also stimulate the release of bioactive compounds, including phenolics, flavonoids, and peptides, which exhibit potent health-promoting properties. This review underscores the transformative potential of processing technologies in enhancing millets as functional ingredients in modern diets, promoting health and advancing sustainable food practices.

Historically, low-moisture foods were considered to have minimal microbial risks. However, they have been linked to many high-profile multistate outbreaks and recalls in recent years, drawing research and extension attention to low-moisture food safety. Limited studies have assessed the food safety research and extension needs for the low-moisture food industry. The objectives of this needs assessment were to explore the food safety culture and education needs, identify the food safety challenges and data gaps, and understand the barriers to adopting food-safety-enhancing technologies in the U.S. low-moisture food industry. This needs assessment was composed of two studies. In Study 1, food safety experts from the low-moisture food industry upper management participated in online interviews and a debriefing discussion session. In Study 2, an online anonymous survey was disseminated to a different group of experts with experience in the low-moisture food industry. The qualitative data were analyzed using deductive and inductive coding approaches, while the quantitative data were analyzed via descriptive analysis. Twenty-five experts participated in the studies (Study 1: n = 12; Study 2: n = 13). Common commodities that participants had worked with included nuts and seeds, spices, flour, and dried fruits and vegetables. A food safety culture conceptual framework was adapted, which included three main components: infrastructure conditions (foundation), individual's food safety knowledge, attitudes, and risk perceptions; and organizational conditions (supporting pillars). Major barriers to establishing a positive food safety culture were identified to be limited resources, difficulties in risk communication, and difficulties in behavioral change. For continual improvement in food safety performance, two major themes of food safety challenges and data gaps were identified: cleaning, sanitation, and hygienic design; and pathogen reduction. Participants perceived the main barriers discouraging the low-moisture food industry from adopting food-safety-enhancing technologies were: (1) budgetary priorities, (2) operation constraints, (3) technology validation, (4) consumer acceptance, and (5) maintaining desired product characteristics such as quality and sensory functionality. The findings of this needs assessment provide guidance for the food industry, academia, and government agencies about the direction of future research and the development of targeted extension programs that might help improve food safety in the low-moisture food industry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Glioblastoma is a Grade 4 primary brain tumor defined by therapy resistance, diffuse infiltration, and near-uniform lethality. The underlying mechanisms are unknown, and no treatment has been curative. Using a recently developed creatine kinase inhibitor (CKi), we explored the role of this inhibitor on GBM biology in vitro. While CKi minimally impacted GBM cell proliferation and viability, it significantly affected migration. In established GBM cell lines and patient-derived xenografts, CKi ablated both the migration and invasion of GBM cells. CKi also hindered radiation-induced migration. RNA-seq revealed a decrease in invasion-related genes, with an unexpected increase in glutathione metabolism and ferroptosis protection genes post-CKi treatment. The effects of CKi could be reversed by the addition of cell-permeable glutathione. Carbon-13 metabolite tracing indicated heightened glutathione biosynthesis post-CKi treatment. Combinatorial CKi blockade and glutathione inhibition or ferroptosis activation abrogated cell survival. Our data demonstrated that CKi perturbs promigratory and anti-ferroptotic roles in GBM, identifying the creatine kinase axis as a druggable target for GBM treatment., (© 2024. The Author(s).)

This study was conducted to evaluate the effects of relative humidity (RH) on moisture loss and flavor in dry-aged beef. Sixteen strip loins were assigned to one of the four aging treatments: vacuum (WET), dry-aging at 50% RH, dry-aging at 70% RH, or dry-aging at 85% RH and aged for 42 days at 2 °C. Loins were evaluated for evaporation loss, trim loss, tenderness, sensory, and microbiological characteristics. Results show that lower RH results in accelerated moisture loss during the first 3 days of the aging process without significantly affecting the total amount of moisture loss. Pseudomonadales dominated the aerobically dry-aged loins while Enterobacteriales was the most abundant in the wet-aged samples. Dry-aged samples had increased content of free amino acids in the cooked meat juice compared to the wet-aged counterpart. Dry aging at 50% RH tended to associate with more desirable flavor notes., Competing Interests: Declaration of competing interest None. There are no potential conflicts., (Published by Elsevier Ltd.)

Current influenza vaccine is not effective in providing cross-protection against variants. We evaluated the immunogenicity and efficacy of multi-subtype neuraminidase (NA) and M2 ectodomain virus-like particle (m-cNA-M2e VLP) and chimeric M2e-H3 stalk protein vaccines (M2e-H3 stalk) in ferrets. Our results showed that ferrets with recombinant m-cNA-M2e VLP or M2e-H3 stalk vaccination induced multi-vaccine antigen specific IgG antibodies (M2e, H3 stalk, NA), NA inhibition, antibody-secreting cells, and IFN-γ secreting cell responses. Ferrets immunized with either m-cNA-M2e VLP or M2e-H3 stalk vaccine were protected from H1N1 and H3N2 influenza viruses by lowering viral titers in nasal washes, trachea, and lungs after challenge. Vaccinated ferret antisera conferred broad humoral immunity in naïve mice. Our findings provide evidence that immunity to M2e and HA-stalk or M2e plus multi-subtype NA proteins induces cross-protection in ferrets., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Millets are rich in nutritional and bioactive compounds, including polyphenols and flavonoids, and have the potential to combat malnutrition and various diseases. However, extracting these bioactive compounds can be challenging, as conventional methods are energy-intensive and can lead to thermal degradation. Green-assisted techniques have emerged as promising methods for sustainable and efficient extraction. This review explores recent trends in employing green-assisted techniques for extracting bioactive compounds from millets, and potential applications in the food and pharmaceutical industries. The objective is to evaluate and comprehend the parameters involved in different extraction methods, including energy efficiency, extraction yield, and the preservation of compound quality. The potential synergies achieved by integrating multiple extraction methods, and optimizing extraction efficiency for millet applications are also discussed. Among several, Ultrasound and Microwave-assisted extraction stand out for their rapidity, although there is a need for further research in the context of minor millets. Enzyme-assisted extraction, with its low energy input and ability to handle complex matrices, holds significant potential. Pulsed electric field-assisted extraction, despite being a non-thermal approach, requires further optimization for millet-specific applications, are few highlights. The review emphasizes the importance of considering specific compound characteristics, extraction efficiency, purity requirements, and operational costs when selecting an ideal technique. Ongoing research aims to optimize novel extraction processes for millets and their byproducts, offering promising applications in the development of millet-based nutraceutical food products. Therefore, the current study benefits researchers and industries to advance extraction research and develop efficient, sustainable, and scalable techniques to extract bioactive compounds from millets., Competing Interests: The authors declare no competing interests associated with the research, ensuring the impartiality and integrity of the presented findings., (© 2024 The Authors.)

A surrogate is commonly used for process validations. The industry often uses the target log cycle reduction for the test (LCR Test ) microorganism (surrogate) to be equal to the desired log cycle reduction for the target (LCR Target ) microorganism (pathogen). When the surrogate is too conservative with far greater resistance than the pathogen, the food may be overprocessed with quality and cost consequences. In aseptic processing, the Institute for Thermal Processing Specialists recommends using relative resistance (D Target )/(D Test ) to calculate LCR Test (product of LCR Target and relative resistance). This method uses the mean values of D Target and D Test and does not consider the estimating variability. We defined kill ratio (KR) as the inverse of relative resistance.The industry uses an extremely conservative KR of 1 in the validation of food processes for low-moisture foods, which ensures an adequate reduction of LCR Test , but can result in quality degradation. This study suggests an approach based on bootstrap sampling to determine conservative KR, leading to practical recommendations considering experimental and biological variability in food matrices. Previously collected thermal inactivation kinetics data of Salmonella spp. (target organism) and Enterococcus faecium (test organism) in Non-Fat Dried Milk (NFDM) and Whole Milk Powder (WMP) at 85, 90, and 95°C were used to calculate the mean KR. Bootstrapping was performed on mean inactivation rates to get a distribution of 1000 bootstrap KR values for each of the treatments. Based on minimum temperatures used in the industrial process and acceptable level of risk (e.g., 1, 5, or 10% of samples that would not achieve LCR Test ), a conservative KR value can be estimated. Consistently, KR increased with temperature and KR for WMP was higher than NFDM. Food industries may use this framework based on the minimum processing temperature and acceptable level of risk for process validations to minimize quality degradation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Radiofrequency (RF) heating has been extensively studied for pasteurizing low-moisture foods. Currently, bulk foods are treated with radiofrequency; potential cross-contamination may occur during packaging of pasteurized products. As an alternative, in-package RF processing was evaluated for Salmonella inactivation on black peppercorns and dried basil leaves and prevention of cross-contamination during storage postprocessing. In-package steaming refers to the process in which the samples were heated in a steam vent package to generate and retain steam during the treatment. This treatment achieved good heating uniformity which could be because of the circulation of steam within the package. One-way steam vent allowed the release of excess steam once a threshold pressure was achieved and later returned to its original position to seal the package, when the RF energy was removed. In-package RF steaming of black peppercorns and dried basil leaves for 135 s and 40 s, respectively, resulted in more than 5 log reduction of Salmonella. The steam vent remained stable posttreatment and properly sealed the package to protect the product from any external contamination. These results indicate that the use of steam vent could effectively pasteurize black peppercorns and dried basil leaves could be beneficial in preventing the potential cross-contamination postprocessing., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Gut microbiota plays a crucial role in inflammatory bowel diseases (IBD) and can potentially prevent IBD through microbial-derived metabolites, making it a promising therapeutic avenue. Recent evidence suggests that despite an unclear underlying mechanism, red cabbage juice (RCJ) alleviates Dextran Sodium Sulfate (DSS)-induced colitis in mice. Thus, the study aims to unravel the molecular mechanism by which RCJ modulates the gut microbiota to alleviate DSS-induced colitis in mice. Using C57BL/6J mice, we evaluated RCJ's protective role in DSS-induced colitis through two cycles of 3% DSS. Mice were daily gavaged with PBS or RCJ until the endpoint, and gut microbiota composition was analyzed via shotgun metagenomics. RCJ treatment significantly improved body weight ( p ≤ 0.001), survival in mice ( p < 0.001) and reduced disease activity index (DAI) scores. Further, RCJ improved colonic barrier integrity by enhancing the expression of protective colonic mucins ( p < 0.001) and tight junction proteins ( p ≤ 0.01) in RCJ + DSS-treated mice compared to the DSS group. Shotgun metagenomic analysis revealed an enrichment of short-chain fatty acids (SCFAs)-producing bacteria ( p < 0.05), leading to increased Peroxisome Proliferator-Activated Receptor Gamma (PPAR-γ) activation ( p ≤ 0.001). This, in turn, resulted in repression of the nuclear factor κB (NFκB) signaling pathway, causing decreased production of inflammatory cytokines and chemokines. Our study demonstrates colitis remission in a DSS-induced mouse model, showcasing RCJ as a potential modulator for gut microbiota and metabolites, with promising implications for IBD prevention and treatment.

With concerns about the efficacy of repeat annual influenza vaccination, it is important to better understand the impact of priming vaccine immunity and develop an effective vaccination strategy. Here, we determined the impact of heterologous prime-boost vaccination on inducing broader protective immunity compared to repeat vaccination with the same antigen. The primed mice that were intramuscularly boosted with a heterologous inactivated influenza A virus (H1N1, H3N2, H5N1, H7N9, H9N2) vaccine showed increased strain-specific hemagglutination inhibition titers against prime and boost vaccine strains. Heterologous prime-boost vaccination of mice with inactivated viruses was more effective in inducing high levels of IgG antibodies specific for groups 1 and 2 hemagglutinin stalk domains, as well as cross-protection, compared to homologous vaccination. Both humoral and T cell immunity were found to play a critical role in conferring cross-protection by heterologous prime-boost vaccination. These results support a strategy to enhance cross-protective efficacy by heterologous prime-boost influenza vaccination.

Background: Current influenza vaccines deliver satisfactory results in young people but are less effective in the elderly. Development of vaccines for an ever-increasing aging population has been an arduous challenge due to immunosenescence that impairs the immune response in the aged, both quantitatively and qualitatively., Results: To potentially enhance vaccine efficacy in the elderly, we investigated the immunogenicity and cross-protection of influenza hemagglutinin virus-like particles (HA-VLP) incorporated with glycosylphosphatidylinositol (GPI)-anchored cytokine-adjuvants (GPI-GM-CSF and GPI-IL-12) via protein transfer in aged mice. Lung viral replication against homologous and heterologous influenza viruses was significantly reduced in aged mice after vaccination with cytokine incorporated VLPs (HA-VLP-Cyt) in comparison to HA-VLP alone. Enhanced IFN-γ + CD4 + and IFN-γ + CD8 + T cell responses were also observed in aged mice immunized with HA-VLP-Cyt when compared to HA-VLP alone., Conclusions: Cytokine-adjuvanted influenza HA-VLP vaccine induced enhanced protective response against homologous influenza A virus infection in aged mice. Influenza HA-VLP vaccine with GPI-cytokines also induced enhanced T cell responses correlating with better protection against heterologous infection in the absence of neutralizing antibodies. The results suggest that a vaccination strategy using cytokine-adjuvanted influenza HA-VLPs could be used to enhance protection against influenza A virus in the elderly., (© 2023. The Author(s).)

Bismuth-based perovskites are an important class of materials in the fabrication of lead-free perovskite solar cells. Bi-based Cs 3 Bi 2 I 9 and CsBi 3 I 10 perovskites are getting much attention due to their appropriate bandgap values of 2.05 eV and 1.77 eV, respectively. However, the device optimisation process plays a key role in controlling the film quality and the performance of perovskite solar cells. Hence, a new strategy to improve crystallization as well as the thin film quality is equally important to develop efficient perovskite solar cells. Herein, an attempt was made to prepare the Bi-based Cs 3 Bi 2 I 9 and CsBi 3 I 10 perovskites via the ligand-assisted re-precipitation approach (LARP). The physical, structural, and optical properties were investigated on perovskite films deposited by the solution process for solar cell applications. Cs 3 Bi 2 I 9 and CsBi 3 I 10 -based perovskite-based solar cells were fabricated using the device architecture of ITO/NiO x /perovskite layer/PC 61 BM/BCP/Ag. The device fabricated with CsBi 3 I 10 showed the best power conversion efficiency (PCE) of 2.3% with an improved fill factor (FF) of 69%, V OC of 0.79 V, and J SC of 4.2 mA cm -2 compared to the Cs 3 Bi 2 I 9 -based device which showed a PCE of 0.7% with a FF of 47%, V OC of 0.62 V and J SC of 2.4 mA cm -2 ., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Radiofrequency (RF) assisted thermal processing can significantly enhance the gel firmness of egg white powder compared to the traditional hot room (HR) processing. Thus, the present study aims to delineate the impact of RF processing on the proteins' structure and bio-functional properties of egg white protein gels. The secondary protein conformations of egg white proteins exhibited no significant alteration upon RF-assisted thermal processing over traditional HR processing. In-vitro gastrointestinal (GI) digestion of egg white gels demonstrated that the RF processing did not compromise the accessibility of digestive proteases despite a more robust gel network. Peptides from the GI digest of egg white gel showed that Ovalbumin and Ovotransferrin were the parent proteins of most of the unique peptides generated, and minor structural differences accounted for these peptides. The bioavailability of the egg protein-derived peptides remains unaffected after RF processing without compromising the viability and integrity of the GI epithelial cells., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Abstract: This multi-institutional study assessed the efficacy of Enterococcus faecium NRRL B-2354 as a nonpathogenic Salmonella surrogate for thermal processing of nonfat dry milk powder, peanut butter, almond meal, wheat flour, ground black pepper, and date paste. Each product was analyzed by two laboratories (five independent laboratories total), with the lead laboratory inoculating (E. faecium or a five-strain Salmonella enterica serovar cocktail of Agona, Reading, Tennessee, Mbandaka, and Montevideo) and equilibrating the product to the target water activity before shipping. Both laboratories subjected samples to three isothermal treatments (between 65 and 100°C). A log-linear and Bigelow model was fit to survivor data via one-step regression. On the basis of D80°C values estimated from the combined model, E. faecium was more thermally resistant (P < 0.05) than Salmonella in nonfat dry milk powder (DEf-80°C, 100.2 ± 5.8 min; DSal-80°C, 28.9 ± 1.0 min), peanut butter (DEf-80°C, 133.5 ± 3.1 min; DSal-80°C, 57.6 ± 1.5 min), almond meal (DEf-80°C, 34.2 ± 0.4 min; DSal-80°C, 26.1 ± 0.2 min), ground black pepper (DEf-80°C, 3.2 ± 0.8 min; DSal-80°C, 1.5 ± 0.1 min), and date paste (DEf-80°C, 1.5 ± 0.0 min; DSal-80°C, 0.5 ± 0.0 min). Although the combined laboratory D80°C for E. faecium was lower (P < 0.05) than for Salmonella in wheat flour (DEf-80°C, 9.4 ± 0.1 min; DSal-80°C, 10.1 ± 0.2 min), the difference was ∼7%. The zT values for Salmonella in all products and for E. faecium in milk powder, almond meal, and date paste were not different (P > 0.05) between laboratories. Therefore, this study demonstrated the impact of standardized methodologies on repeatability of microbial inactivation results. Overall, E. faecium NRRL B-2354 was more thermally resistant than Salmonella, which provides support for utilizing E. faecium as a surrogate for validating thermal processing of multiple low-moisture products. However, product composition should always be considered before making that decision., (Published 2022 by the International Association for Food Protection. Not subject to U.S. Copyright.)

Heterologous prime-boost immunization regimens using various vaccine platforms demonstrated promising results against infectious diseases. Here, mice were sequentially immunized with the recombinant baculovirus (rBV), virus-like particle (VLP), and recombinant vaccinia virus (rVV) vaccines expressing the Plasmodium berghei apical membrane antigen 1 (AMA1) for protective efficacy evaluation. The rBV&#95;V&#95;rVV heterologous immunization regimen elicited high levels of parasite-specific IgG, IgG2a, and IgG2b antibody responses in sera. Upon P. berghei challenge infection, proliferations of germinal center B cells in the inguinal lymph nodes, as well as blood CD4 + and CD8 + T cells were induced. More importantly, rBV&#95;V&#95;rVV immunization significantly diminished the parasitemia and prevented drastic bodyweight loss in mice post-challenge infection with P. berghei . Our findings revealed that immunization with rBV, VLP, and rVV expressing the AMA1 conferred protection against P. berghei infection, providing evidence for the potential implementation of this strategy.

To improve cross-protection of influenza vaccination, we tested conjugation of conserved M2e epitopes to the surface of inactivated influenza virus (iPR8-M2e*). Treatment of virus with chemical cross-linker led to diminished hemagglutination activity and failure to induce hemagglutination inhibiting antibodies. Conjugated iPR8-M2e* vaccine was less protective against homologous and heterosubtypic viruses, despite the induction of virus-specific binding IgG antibodies. In alternative approaches to enhance cross-protection, we developed a genetically linked chimeric protein (M2e-B stalk) vaccine with M2e of influenza A and hemagglutinin (HA) stalk of influenza B virus. Vaccination of mice with inactivated influenza A virus supplemented with M2e-B stalk effectively induced hemagglutination inhibiting antibodies, humoral and cellular M2e immune responses, and enhanced heterosubtypic protection. This study demonstrates the importance of HA functional integrity in influenza vaccine efficacy and that supplementation of influenza vaccines with M2e-B stalk protein could be a feasible strategy of improving cross-protection against influenza viruses., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Annual influenza vaccination is recommended to update the variable hemagglutinin antigens. Here, we first designed a virus-like particle (VLP) displaying consensus multi-neuraminidase (NA) subtypes (cN1, cN2, B cNA) and M2 ectodomain (M2e) tandem repeat (m-cNA-M2e VLP). Vaccination of mice with m-cNA-M2e VLP induced broad NA inhibition (NAI), and M2e antibodies as well as interferon-gamma secreting T cell responses. Mice vaccinated with m-cNA-M2e VLP were protected against influenza A (H1N1, H5N1, H3N2, H9N2, H7N9) and influenza B (Yamagata and Victoria lineage) viruses containing substantial antigenic variations. Protective immune contributors include cellular and humoral immunity as well as antibody-dependent cellular cytotoxicity. Furthermore, comparable cross protection by m-cNA-M2e VLP vaccination was induced in aged mice. This study supports a novel strategy of developing a universal vaccine against influenza A and B viruses potentially in both young and aged populations by inducing multi-NA subtype and M2e immunity with a single VLP entity., Competing Interests: The authors declare no conflicts of competing interest exits.

Adjuvants can increase the magnitude and durability of the immune response generated by the vaccine antigen. Aluminum salts (Alum) remain the main adjuvant licensed for human use. A few new adjuvants have been licensed for use in human vaccines since the 1990s. QS-21, a mixture of saponin compounds, was included in the AS01-adjuvanted Shingrix vaccine. Here, we investigated the adjuvant effects of VSA-1, a newly developed semisynthetic analog of QS-21, on promoting protection in mice after vaccination with the inactivated split virus vaccine. The adjuvant effects of VSA-1 on improving vaccine efficacy after prime immunization were evident as shown by significantly higher levels of hemagglutination-inhibiting antibody titers and enhanced homologous protection compared to those by QS-21 and Alum adjuvants. The adjuvant effects of VSA-1 on enhancing heterosubtypic protection after two doses of adjuvanted vaccination were comparable to those of QS-21. T cell immunity played an important role in conferring cross-protection by VSA-1-adjuvanted vaccination. Overall, the findings in this study suggest that VSA-1 exhibits desirable adjuvant properties and a unique pattern of innate and adaptive immune responses, contributing to improved homologous and heterosubtypic protection by inactivated split influenza vaccination in mice.

Passive thermoregulation is an important strategy to prevent overheating in thermally challenging environments. Can the diversity of optical properties found in Christmas beetles (Rutelinae) be an advantage to keep cool? We measured changes in temperature of the elytra of 26 species of Christmas beetles, exclusively due to direct radiation from a solar simulator in visible (VIS: 400-700 nm) and near infrared (NIR: 700-1700 nm) wavebands. Then, we evaluated if the optical properties of elytra could predict their steady state temperature and heating rates, while controlling for size. We found that higher absorptivity increases the heating rate and final steady state of the beetle elytra in a biologically significant range (3 to 5°C). There was substantial variation in the absorptivity of Christmas beetle elytra; and this variation was achieved by different combinations of reflectivity and transmissivity in both VIS and NIR. Size was an important factor predicting the change in temperature of the elytra after 5 min (steady state) but not maximum heating rate. Lastly, we show that the presence of the elytra covering the body of the beetle can reduce heating of the body itself. We propose that beetle elytra can act as a semi-insulating layer to enable passive thermoregulation through high reflectivity of elytra, resulting in low absorptivity of solar radiation. Alternatively, if beetle elytra absorb a high proportion of solar radiation, they may reduce heat transfer from the elytra to the body through behavioral or physiological mechanisms., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology.)