Vaccination

The importance of global vaccines for various infectious agents has been emphasized through our experience with the current COVID-19 vaccine. Physicians trained in allergy and immunology are considered experts in vaccines and adverse reactions from vaccines. This chapter will emphasize how to approach vaccines for various populations, including immunodeficient patients, how to determine best paths for patients who miss vaccine doses, and how to address adverse reactions to vaccines, as noted in Case 2, where the patient experienced an anaphylactic reaction to the measles, mumps, and rubella (MMR) vaccine. In addition, mechanisms behind adverse vaccine reactions are discussed. © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Switzerland AG 2022.

A Functional Assay to Measure the T Cell Response to Sars-Cov-2 in Primary Immunodeficiency Patients

Background/Aims Effective symptom management may require the use of medications. Medication adherence may be hindered by formulation aspects, such as poor taste. Paediatric studies indicate, that despite concerns of swallowing solid dose forms, children prefer these to liquid forms. They find the solid dose forms more palatable. However, swallowing numerous solid dose forms, may present a significant 'pill' burden to patients and their care-givers. Filling empty gelatine capsules with requisite medications is seen and used as a way to address palatability, decrease pill burden and thereby increase compliance. Yet there is little evidence on the impact this practise may have on the effectiveness of over-encapsulated medicines. This study explored the effect of over-encapsulation on in vitro disintegration and dissolution, of some commonly used medicines in paediatric palliative care. Method Immediate release (Cyclizine Hydrochloride, Gabapentin, Paracetamol) and modified release preparations (Omeprazole, Diclofenac sodium) were over-encapsulated in size 00 gelatin and HPMC capsules (n=6). Dissolution and disintegration were tested according to Pharmacopeia standards. Statistical analyses, using Student's T-test and f1 and f2 tests (respectively) were applied to determine similarities or differences in disintegration or dissolution. Results Disintegration and dissolution was prolonged for all over-encapsulated immediate release preparations, especially when using HPMC capsules. However, percentage of drug dissolved met the acceptance criteria for immediate-release solid oral dosage. Over-encapsulation of modified release preparations did not lead to significant dissolution or disintegration changes. Conclusion Over-encapsulation, may delay medication release, especially for immediate release medicines however, medicine effectiveness may not be. Further studies are required before we can safely recommend use of over-encapsulation as an administration compliance aid.

Ultradiluted SARS-CoV-2 Spike Protein mitigates hyperinflammation in lung via ferritin and MMP-9 regulation in BALB/c mice.

AIM: This study investigated the prophylactic and therapeutic role of ultradiluted preparation of the Delta variant of SARS-CoV-2 recombinant spike (S) protein during S antigen-induced inflammatory process of disease progression along with the probable mechanism of action. MAIN METHODS: Ultradiluted S protein (UDSP) was prepared and administered orally to adult BALB/c mice before and after administration of S antigen intranasally. After an observation period of 72 h, animals were sacrificed and expression level of ferritin was assayed through ELISA. The genetic expressions of cytokines, IL-6, IL-10, IL-1ß, TNFα, IL-17, MMP-9, TIMP-1, ferritin light and heavy chains, and mitochondrial ferritin from lung tissues were investigated through RT-PCR. Formalin-fixed lung tissue sections were stained with hematoxylin and eosin to observe the degree of pathological changes. The activity of MMP-9 in lung tissues was investigated through gelatin zymography and immunofluorescence of MMP-9 in lung tissue sections was performed to revalidate the finding from gelatin zymography. Systems biology approach was used to elucidate a probable pathway where UDSP attenuated the inflammation through the regulation of pro- and anti-inflammatory cytokines. KEY FINDINGS: UDSP attenuated the S antigen-induced hyperinflammation in the lung by regulating pro- and anti-inflammatory cytokines, calming cytokine storm, reducing ferritin level both in transcriptional and translational levels, and restoring critical ratio of MMP-9: TIMP-1. SIGNIFICANCE: Our findings suggest a probable pathway by which UDSP might have attenuated inflammation through the regulation of cytokines, receptors, and other molecules. This proclaims UDSP as a promising antiviral agent in the treatment of COVID-19-induced immunopathogenesis.

Synthesis and characterization of gelatin/lignin hydrogels as quick release drug carriers for Ribavirin.

In this study, hydrogels based on gelatin and lignin were fabricated as efficient drug carriers for Ribavirin. The obtained hydrogels were characterized by scanning electron microscopy (SEM), ATR-FTIR spectroscopy, differential scanning calorimetry (DSC), mechanical compression and rheometry. Results showed that the pore structure, viscoelastic behavior and swelling ability significantly influenced by varying lignin content and crosslinker ratio. By increasing the crosslinker N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC) content, the pore size became smaller, while increasing the lignin content resulted in larger pores. In addition, all hydrogels show strong shear thinning behavior. Ribavirin was used as a drug model, and its release rate was enhanced by increasing lignin content in the binary hydrogel structure. A higher Ribavirin cumulative release was observed for gelatin/lignin with higher lignin content (3 %) hydrogel. These findings emphasize the chemical composition on the structure and the release behavior of lignin-containing hydrogels.

Buriti oil and natural surfactant improve the properties of cassava starch and carboxymethylcellulose films

Due to the environmental problem related to the disposal of non-biodegradable packaging and its aggravation during the COVID-19 pandemic, buriti oil (BO) and Yucca schidigera (YS) surfactant were added to the cassava starch and carboxymethyl-cellulose films to create a new emulsified biofilm with better properties. The influence of BO and YS on mechanical and barrier properties was evaluated. All properties differed significantly (p < 0.05) compared to the base film. Incorporation of BO and YS increased tensile strength by 1.100% and opacity by 375% and reduced elongation. In addition, the two variables generated a reduction in water vapour permeability (similar to 47%) and water solubility (similar to 70%), making the films less hydrophilic. Both attenuated total reflectance (ATR)-Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) were able to generate important correlations, observing the effects of BO and YS on the uniformity and cohesion of the structures of the emulsified films. Overall, the emulsified films showed great potential for application as primary and/or secondary packaging.

Topical Platelet-Rich Plasma: A Novel Delivery Method for an Emerging Treatment of Olfactory Dysfunction

Introduction: Olfactory dysfunction (OD) affects more than 3 million US adults. The number of patients with post viral olfactory dysfunction (PVOD) is expected to increase secondary to the worldwide COVID-19 pandemic. Preliminary studies have demonstrated the efficacy of platelet-rich plasma (PRP) in restoration of smell in both animals and humans. To date, human studies have utilized injectable PRP only. We describe our pilot study investigating the use of topical PRP as a novel delivery method for smell restoration and contribute to existing literature demonstrating the promise of PRP as a therapeutic. Method(s): Pilot study from September 2020 to January 2022. Patients >18 years with hyposmia diagnosed via Brief Smell Identification Test (B-SIT) score <8 were included. PRPimpregnated Surgifoam was placed into bilateral olfactory clefts monthly for at least 3 months. Patients completed the B-SIT at baseline and 1 month after each treatment. Result(s): Eight patients underwent at least 3 treatments and completed the B-SIT at month 4. Average age was 56.3 years;mean smell loss duration was 19.3 months. Etiologies included PVOD, post-COVID (5), and idiopathic. Mean change in B-SIT after 3 treatments was +1.06. Of patients, 62.5% had achieved the minimal clinically significant difference of >1 on B-SIT after treatments 2 and 3. Patients with smell loss <12 months demonstrated greater B-SIT scores at 4 months (+1.6 vs +0.2). Two patients achieved B-SIT >8 after 3 treatments. Of patients who have returned thus far for a fourth treatment, 1 additional patient scored >8. Conclusion(s): We present the largest pilot study to date for the use of PRP in treatment of OD and the first study to develop methods for topical delivery in human subjects. Topical PRP may serve as a less invasive, efficacious therapy for patients. Further, randomized control trials are warranted to investigate the required number of topical PRP treatments for smell restoration.

Generation Of Macro-scale And Mature Lung Organoids Through Engineering Cell Microenvironment

Purpose/Objectives: <Most used lower respiratory tract models consist of cell monolayers which lack of tissue and organ level response and of in-vivo phenotype. Ex-vivo lung tissues have short viability and limited availability. Lung organoids, which recapitulates better the 3D cellular complex structures, architecture, and in-vivo function, fail to reach maturity even after 85 -185 days of culture. Therefore, these models have a limited use to study fetal lung diseases. Other lung models, consist of only one structure of the lower track, such as bronchial tubes or alveoli, but fail to recapitulate the whole organ structure. In this work, cell microenvironment was used to promote the self-organization of epithelial and mesenchymal cells into macro-structures, aiming to mimic the whole and adult lower respiratory tract model> Methodology: <Different parts of the microenvironment were considered to create a compliant matrix. Alginate-Gelatin hydrogels were used for 3D encapsulation of mesenchymal origin cells. This hydrogel provided a stiffness like the one on the lung. Base membrane zone proteins were used to induce the attachment and guidance of epithelial cells into 3D structures. The interactions between both cell types, further guided them into lung fate. The morphology of resulting organoids was analyzed using immunostaining and confocal microscopy, LSM710, with the purpose of evaluate polarization, protein markers, and different cell populations. Quantitative PCR was performed to evaluate and compare the expression of lung fate genes with traditional cell monocultures.> Results: <The engineered microenvironment and protocol development done in this work resulted in macro-scale structures, in which branching morphogenesis occurred at day 21. Different structures were identified in the organoid including bronchial tube, bronchioles, and alveoli. Polarization of the organoids was confirmed by visualization of E-cadherin, and ZO-1. Expression of Surfactant Protein B and C into the organoids confirmed the presence of alveolar type II cells, which are only present in the later development stage. Surfactant Protein B, Transmembrane protease, serine 2, TMPRSS-2, and Angiotensin-converting enzyme 2, ACE2 were found to be significantly higher expressed into the organoids in comparison with traditional epithelial cells monolayers.> Conclusion/Significance: <Growth factors are normally used to induce the fate of stem cells into lung organoids;however, these fail to reach maturity. Here, we developed a new methodology to induce the formation of the organoids based on the cell microenvironment. The resulting organoids require less time for development. The initial stage of adult cells can be modulated through culture conditions induce a 3D structure like the adult lung. As such, these organoids have the potential to be used for modeling adult diseases and to develop specific models from patient cells, which is one step forward to personalized medicine. SFTPB is one of the main proteins which facilitates the breathing process. Its high expression into our model may indicate that breathing occurs into our lung organoids. The higher expression of TMPRSS-2 and ACE2 into the organoids has a major significance in the field of virology since both proteins are the mainly entrance of SARS-CoV-2, and influenza H1N1.>.

PCL- And Gelatin-based Electrospun Biological Scaffolds For In Vitro Lung Tissue Engineering

Purpose/Objectives: Acute and chronic respiratory diseases constitute a substantial socioeconomic burden on a global scale, as made abundantly clear in the last two years with the rampant coronavirus disease 2019 (COVID-19) pandemic. Alas, the development of new therapies for pathological respiratory conditions has been hindered by the inadequacy of current preclinical models, which often fail to provide reliable predictions on drug safety and efficacy in humans. In particular, considerable anatomical and physiological differences between the respiratory systems of commonly used animal models and humans are one of the main issues leading to high drug attrition and clinical failure rates. Accordingly, the generation of physiologically relevant preclinical lung models for early drug development and pharmaceutical research is urgently needed. In this work, poly(ϵ-caprolactone) (PCL) and gelatin were used as raw materials to produce electrospun scaffolds for in vitro lung tissue engineering, in order to generate human biomimetic platforms for preclinical drug safety and efficacy testing. Methodology: PCL and gelatin were mixed at varying volume ratios: 1:0 (PP), 6:1 (PPG61), 4:1 (PPG41), and 2:1 (PPG21), so as to determine the optimal gelatin concentration for cell adhesion and growth. Poly(vinylpyrrolidone) (PVP) was added to every polymer mixture to facilitate the electrospinning process, and electrospun fibrous matrices were fabricated using a needleless electrospinning technique. Scaffold morphology, chemical composition, and wettability were characterized with scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, and water contact angle analysis, respectively. Biocompatibility testing was performed using human bronchial (16HBE) and alveolar (A549) epithelial cell lines, consisting of cell metabolic activity, proliferation, and adhesion evaluation over two weeks of in vitro culture. Results: All polymer blends resulted in the formation of electrospun scaffolds with a nanofibrous structure. The addition of gelatin in PPG61 scaffolds improved fiber morphology compared to PP formulations, but increasing proportions of this polymer in PPG41 and PPG21 mats caused a larger number of defects, such as beading and branching. FTIR analysis confirmed the presence of PCL and PVP in PP scaffolds, as well as the addition of gelatin in all PPG blends. Moreover, as expected, all scaffolds were hydrophilic, with water contact angles below 90°, being suitable for protein adsorption and cell adhesion. Regarding 16HBE and A549 cell viability, surprisingly, no major differences were found between the different formulations over the two-week culture period, showing that all polymer blends were equally capable of promoting cell adhesion and growth. While PP scaffolds significantly outperformed PPG electrospun mats in early timepoints, no such differences were identified at the end of the experimental period. Conclusion/Significance: These results suggested that PCL, PVP, and/or gelatin blend electrospun scaffolds are conducive to lung epithelial cell adhesion and proliferation. Nevertheless, further studies investigating epithelial cell differentiation and function should be conducted to fully assess the suitability of these biomaterials as platforms for in vitro lung tissue engineering.

Development of amperometric enzyme-based biosensor to evaluate the adulteration in virgin coconut oil. (Special Issue: Innovative methods in food process engineering.)

Virgin Coconut oil (VCO), owing to its functional properties (important in COVID-19), is costly and, therefore, susceptible to adulteration with other cheaper oils like coconut oil. An enzyme-based biosensor confirmative test of VCO was constructed by co-immobilizing enzymes onto a glassy carbon electrode. The performance of the biosensor was optimized at a potential of +0.5 V with 45 mg gelatin, 30 mg BSA coupled with 2.5% glutaraldehyde at pH 7.0 with an incubation time of 1 hr. Adulterated samples of VCO with coconut oil (CO) were analyzed. The concentration of diglyceride (DG) was estimated from the empirical relation, which showed a linear increase with the increase in adulteration. The developed biosensor was validated using high-performance liquid chromatography (HPLC) methods using a paired sample t test at a 5% significance level. The biosensor could detect adulteration in VCO with CO above 20% within 3-5 s and can be reused for 25 days.

Utilization of By-Products from Livestock: Study on the Mechanical and Thermal Properties of Biodegradable Containers Made with Pork Skin Gelatin Polymer.

This study aimed to develop a biodegradable container made of pork gelatin. Gelatin was extracted from pork skin by hot water at 80 °C, and containers were prepared by adding eggshell powder (20%) as a pore agent, and walnut powder (0.08 wt%; PEW1, 0.14 wt%; PEW2) to improve hardness. The blends were molded for each experiment and dried at 30 °C for 24 h, at 40 °C for 16 h, and at 121 °C for 16 h. The containers were analyzed with respect to morphological (SEM; scanning electron microscope), mechanical (tensile strain and stress), and thermal (DSA; differential scanning calorimetry and TGA; thermogravimetric analysis) properties, as well as biodegradability. SEM investigation showed a smoother surface for PEW1 than for PEW2. The tensile stress of PEW2 (37.86 MPa) was significantly higher than that of PEW1 (28.40 MPa), and the melting enthalpies were 137.60 J/g (PEW1) and 309.60 J/g (PEW2). TGA showed similar properties, but PEW2 contained more lignin; therefore, its decomposition temperature was higher. The PEW1 and PEW2 containers were completely biodegraded after approximately 7 and 11 weeks, respectively. Walnut shell powder increased the hardness, but slowed the biodegradation process. The applications of this biodegradable container are short-lived products such as food packaging.

Biopolymeric sustainable materials and their emerging applications

Advancements in polymer science and engineering have helped the scientific community to shift its attention towards the use of environmentally benign materials for reducing the environmental impact of conventional synthetic plastics. Biopolymers are environmentally benign, chemically versatile, sustainable, biocompatible, biodegradable, inherently functional, and ecofriendly materials that exhibit tremendous potential for a wide range of applications including food, electronics, agriculture, textile, biomedical, and cosmetics. This review also inspires the researchers toward more consumption of biopolymer-based composite materials as an alternative to synthetic composite materials. Herein, an overview of the latest knowledge of different natural- and synthetic-based biodegradable polymers and their fiber-reinforced composites is presented. The review discusses different degradation mechanisms of biopolymer-based composites as well as their sustainability aspects. This review also elucidates current challenges, future opportunities, and emerging applications of biopolymeric sustainable composites in numerous engineering fields. Finally, this review proposes biopolymeric sustainable materials as a propitious solution to the contemporary environmental crisis. © 2022 Elsevier Ltd.

Gelatin Stabilizes Nebulized Proteins in Pulmonary Drug Delivery against COVID-19.

Delivering medication to the lungs via nebulization of pharmaceuticals is a noninvasive and efficient therapy route, particularly for respiratory diseases. The recent worldwide severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) pandemic urges the development of such therapies as an effective alternative to vaccines. The main difficulties in using inhalation therapy are the development of effective medicine and methods to stabilize the biological molecules and transfer them to the lungs efficiently following nebulization. We have developed a high-affinity angiotensin-converting enzyme 2 (ACE2) receptor-binding domain (RBD-62) that can be used as a medication to inhibit infection with SARS-CoV-2 and its variants. In this study, we established a nebulization protocol for drug delivery by inhalation using two commercial vibrating mesh (VM) nebulizers (Aerogen Solo and PARI eFlow) that generate similar mist size distribution in a size range that allows efficient deposition in the small respiratory airway. In a series of experiments, we show the high activity of RBD-62, interferon-α2 (IFN-α2), and other proteins following nebulization. The addition of gelatin significantly stabilizes the proteins and enhances the fractions of active proteins after nebulization, minimizing the medication dosage. Furthermore, hamster inhalation experiments verified the feasibility of the protocol in pulmonary drug delivery. In short, the gelatin-modified RBD-62 formulation in coordination with VM nebulizer can be used as a therapy to cure SARS-CoV-2.

BIOFABRICATION OF NANOSTRUCTURED GELATIN FIBER VIA CENTRIFUGAL SPINNING METHOD

In this study, type-I bovine collagen solved in diluted acidic solutions (acidic acid and formic acid) and fed into the centrifugal spinning device to obtain nanofiber formation. The centrifugal spinning device is providing submicron fibers thanks to its rotational movement during the solution feeding. Different feeding ratios and rotational speeds are studied to optimize the process. Obtained na nofiber webs were observed via scanning electron microscopy (SEM). Structural characterization of samples was investigated via FTIR. Fabricated collagen-based nanofibers filtration performance was evaluated in terms of filtration efficiency and air permeability tests. Tests results are pointed that fabricated gelatin nanofibers can be an efficient alternative biomaterial design against Covid-19.

Berry By-Products in Combination with Antimicrobial Lactic Acid Bacteria Strains for the Sustainable Formulation of Chewing Candies

The purpose of this research was to develop formulations of chewing candies (CCs) in a sustainable manner by using berry by-products in combination with antimicrobial lactic acid bacteria (LAB) strains. To implement this aim, the optimal quantities of by-products from lyophilised raspberry (Rasp) and blackcurrant (Bcur) from the juice production industry were selected. Prior to use, Lactiplantibacillus plantarum LUHS135, Liquorilactobacillus uvarum LUHS245, Lacticaseibacillus paracasei LUHS244, and Pediococcus acidilactici LUHS29 strains were multiplied in a dairy industry by-product—milk permeate (MP). The antimicrobial activity of the selected ingredients (berry by-products and LAB) was evaluated. Two texture-forming agents were tested for the CC formulations: gelatin (Gl) and agar (Ag). In addition, sugar was replaced with xylitol. The most appropriate formulation of the developed CCs according to the product’s texture, colour, total phenolic compound (TPC) content, antioxidant activity, viable LAB count during storage, overall acceptability (OA), and emotions (EMs) induced in consumers was selected. It was established that the tested LAB inhibited three pathogens out of the 11 tested, while the blackcurrant by-products inhibited all 11 tested pathogens. The highest OA was shown for the CC prepared with gelatin in addition to 5 g of Rasp and 5 g of Bcur by-products. The Rasp and LUHS135 formulation showed the highest TPC content (147.16 mg 100 g−1 d.m.), antioxidant activity (88.2%), and LAB count after 24 days of storage (6.79 log10 CFU g−1). Finally, it was concluded that Gl, Rasp and Bcur by-products, and L. plantarum LUHS135 multiplied in MP are promising ingredients for preparing CCs in a sustainable manner;the best CC formula consisted of Gl, Rasp by-products, and LUHS135 and showed the highest OA (score 9.52) and induced the highest intensity of the EM ‘happy’ (0.231).

State-of-the-art review of advanced electrospun nanofiber yarn-based textiles for biomedical applications.

The pandemic of the coronavirus disease 2019 (COVID-19) has made biotextiles, including face masks and protective clothing, quite familiar in our daily lives. Biotextiles are one broad category of textile products that are beyond our imagination. Currently, biotextiles have been routinely utilized in various biomedical fields, like daily protection, wound healing, tissue regeneration, drug delivery, and sensing, to improve the health and medical conditions of individuals. However, these biotextiles are commonly manufactured with fibers with diameters on the micrometer scale (> 10 µm). Recently, nanofibrous materials have aroused extensive attention in the fields of fiber science and textile engineering because the fibers with nanoscale diameters exhibited obviously superior performances, such as size and surface/interface effects as well as optical, electrical, mechanical, and biological properties, compared to microfibers. A combination of innovative electrospinning techniques and traditional textile-forming strategies opens a new window for the generation of nanofibrous biotextiles to renew and update traditional microfibrous biotextiles. In the last two decades, the conventional electrospinning device has been widely modified to generate nanofiber yarns (NYs) with the fiber diameters less than 1000 nm. The electrospun NYs can be further employed as the primary processing unit for manufacturing a new generation of nano-textiles using various textile-forming strategies. In this review, starting from the basic information of conventional electrospinning techniques, we summarize the innovative electrospinning strategies for NY fabrication and critically discuss their advantages and limitations. This review further covers the progress in the construction of electrospun NY-based nanotextiles and their recent applications in biomedical fields, mainly including surgical sutures, various scaffolds and implants for tissue engineering, smart wearable bioelectronics, and their current and potential applications in the COVID-19 pandemic. At the end, this review highlights and identifies the future needs and opportunities of electrospun NYs and NY-based nanotextiles for clinical use.

Cystatin C and Neutrophil Gelatin-associated Lipocalin (NGAL) Can Predict Acute Kidney Injury and In-Hospital Mortality in COVID-19 Patients

Background: Prediction and early diagnosis of acute kidney injury (AKI) in critically ill Coronavirus disease 2019 (COVID-19) patients are of great importance. Therefore, using promising renal biomarkers such as cystatin C and neutrophil gelatinase-associated lipocalin (NGAL) to identify the risk of future AKI is crucial. Materials and Methods: A total of 89 adult patients with COVID-19 were included in this study. Serum cystatin C and NGAL concentration were assessed on intensive care unit (ICU) admission then repeated after 48 hours. Serum creatinine was followed for 7 days to report the development of AKI. Results: Among the COVID-19 patients, 28.1% developed AKI. Although admission serum creatinine was not significantly different between the AKI group and the non-AKI group (p=0.375), admission Cystatin C (p=0.018), and NGAL (p<0.001) were significantly different between both groups. After 48 hours, a change in Cystatin C level (p<0.001) but not NGAL (p=0.4) was a predictor for AKI. Logistic regression model including age (p=0.031), Cystatin C on 48 hrs (p=0.003) and NGAL on admission (p=0.015) could predict AKI in COVID-19 patients. Conclusion: Serum Cystatin C and NGAL in ICU could be used to predict AKI in COVID-19 patients. A logistic regression model including age, Cystatin C on 48hrs, and NGAL on admission might be a tool for individualized risk estimation of AKI in COVID-19 patients.

Phenotypic and Genomic Analysis of Novel, Fastidious, Gram-negative Bacilli Isolated from Clinical Wound Specimens

Background. Animal bites are considered the thirteenth leading cause of nonfatal ED visits. Epidemiology studies have shown a rise in dog bites during the COVID-19 pandemic in the U.S. In Oct. 2020, we received a facultatively anaerobic, non-hemolytic Gram-negative rod (OL1) from a dog bite wound for identification. 16S rRNA gene sequencing showed OL1 was 95.9% identical to Ottowia pentelensis in the family Comamonadaceae. Our historical sequence database revealed 8 additional isolates (OL2-OL9) from hand wounds/abscesses (including 3 dog bites) since 2012 that had > 99.8% identity with OL1. Most other Ottowia sp. have been isolated from industrial and food sources, with no reports from patient samples. As these clinical isolates likely represent a novel Ottowia species, we aimed to characterize them using both phenotypic and genomic approaches. Methods. The OL isolates were tested in API 20 NE panels (8 conventional and 12 assimilation tests) for 4 d. Paired-end genomic DNA libraries (Nextera DNA Flex Library Prep, Illumina) were sequenced as 150 nt reads by Illumina NovaSeq. De novo assembly, annotation, functional prediction, and phylogenetic analyses were performed with Geneious, PATRIC, and web-prediction databases. Strain comparison was done with StrainTypeMer. Results. All 9 OL isolates were negative for indole, urea, arginine, esculin, PNPG, glucose fermentation and carbohydrate assimilation tests. Potassium gluconate assimilation and gelatin hydrolysis were positive for 5 and 4 isolates, respectively. StrainTypeMer showed the isolates from different patients were not closely related, but 2 from the same patient were indistinguishable. The estimated genome size was ~3.1 Mbp, with 66.1% G/C, and ~3523 coding genes. Potential virulence factors (BrkB and MviM), multidrug efflux systems (MdtABC-TolC and Bcr/CflA), and 1-2 intact prophages were identified. Genomic phylogenetic analysis with RAxML showed the OL isolates clustered separately from all known Ottowia spp. Conclusion. These OL isolates are fastidious, Gram-negative bacilli from clinical wound specimens, and are associated with dog bites. Genomic and 16S rRNA gene sequence analysis suggests these isolates constitute a novel species within the family Comamonadaceae.

Variation in Severe Acute Respiratory Syndrome Coronavirus 2 Bioaerosol Production in Exhaled Breath.

We developed a simple, noninvasive mask sampling method to quantify and sequence severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from exhaled breath. We found substantial variation between individuals in SARS-CoV-2 copies exhaled over a 15-minute period, which moderately correlated with nasal swab viral load. Talking was associated with a median of 2 log10 greater exhaled viral copies. Exposure varies substantially between individuals but may be risk stratified by nasal swab viral load and whether the exposure involved conversation.

Quality assessment of honey in three different geographical areas from Republic of Moldova

One of the empirical and image products of the Republic of Moldova is bee honey. Its assortment is due to the variety of landforms, as well as the diversity of flora specific to geographical regions. During the Covid-19 pandemic, domestic consumers had limited access to bee honey. This was caused by the restrictions imposed in order to organize fairs and agricultural markets which are one of the main sources for the purchase of bee products in the Republic of Moldova. At the same time, the analysis of trademarks in supermarkets highlighted the preferences of honey consumption as follows: polyfloral honey-28.57%, lime honey-20.40% and acacia honey-14.28%. In order to evaluate the quality of honey from small producers and highlight the specific characteristics of geographical areas, 60 samples were analyzed that included 3 types of honey: polyflora, sunflower and linden collected from 3 different geographic areas: Soroca area (North), area Ungheni (Center), Stefan Voda area (South). Honey samples were declared harvest of 2020. They were analyzed physico-chemically using methods provided by national and EU standards. The results obtained from the analysis of pollen in honey confirm the botanical origin declared by beekeepers and allowed to highlight the types of pollen specific to each area. Following the determination of qualitative indices: reaction with ethyl alcohol, with resorcinol;insoluble matter, cereal flour, gelatin and starch, it was found that the most inconsistencies were found in linden honey. Some indicators were close to the maximum permissible values specified in regulatory documents. The water content found in the honey samples was quite varied, and ranged from 16.05% +or- 0.11% to 19.89% +or- 0.13%, but these samples were within the limits of the standards. Total acidity ranged from 6.19 +or- 0.13 to 27.20 +or- 0.03 which falls within the established norms (up to 50.00 cm3 NaOH solution in (milliequivalents) per 100 g of honey). According to the SIE Lab space, all samples have a yellowish tint, this is indicated by positive values on the b* axis. However, honey samples from the southern region of Moldova showed the presence of greenish pollen, as evidenced by the reduced values on the a* axis. Our research reveals the quality level of honey from three different areas of the Republic of Moldova and the variation of quality parameters due to factors such as geographical region, climatic conditions, botanical origin and handling or storage conditions.

Effect of hydroxychloroquine sulfate on the gelation behavior, water mobility and structure of gelatin.

Hydroxychloroquine sulfate (HCQ) is a well-established antimalarial drug that has received considerable attention during the COVID-19 associated pneumonia epidemic. Gelatin is a multifunctional biomacromolecule with pharmaceutical applications and can be used to deliver HCQ. The effect of HCQ on the gelation behaviors, water mobility, and structure of gelatin was investigated to understand the interaction between the drug and its delivery carrier. The gel strength, hardness, gelling (Tg) and melting (Tm) temperatures, gelation rate (kgel), and water mobility of gelatin decreased with increasing amounts of HCQ. The addition of HCQ led to hydrogen bonding that interfered with triple helix formation in gelatin. Fourier transform infrared spectroscopy (FTIR) and X-ray diffractometer (XRD) analysis further confirmed that the interaction between HCQ and gelatin is primarily through hydrogen bonding. Atomic force microscopy (AFM) revealed that higher content of HCQ resulted in more and larger aggregates in gelatin. These results provide not only an important understanding of gelatin for drug delivery design but also a basis for the studying interactions between a drug and its delivery carrier.