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In present work, hydrophobic copper(I) oxide (Cu2O) surfaces were fabricated by chemical bath deposition method. Copper oxide layers on glass slides were coated using copper sulfate as a precursor. To examine wetting properties of copper oxide surfaces, various samples were prepared at different numbers of coating cycles and sintering conditions (temperature, time, and atmosphere). Morphology, composition, and optical absorption of the copper oxide layers were characterized by profilometer, AFM, XRD, UV-Vis photospectroscopy, and water contact angle measurements. It was observed that hydrophobicity decreases by increasing oxygen amount of the sintering atmosphere. In fact, this is the result of an increase in the surface oxygen amount and consequently the increase of surface energy. An optimum thickness and surface morphology is obtained for hydrophobicity of these thin films which are due to air trapping at more narrow valleys and development of the Cassie-Baxter phase. Moreover, the effects of valley's height and width on the wetting were investigated. It is shown that the width of the valleys is a more important factor in developing the Cassie-Baxter phase than the height of valleys. Furthermore, the obtained results show that all copper(І) oxide surfaces tend to a hydrophobicity behavior after 1 week drying at ambient conditions. The measured water contact angles of Cu2O layers were as high as 112°, without sintering or fatty acid modification. Nevertheless, it was enhanced up to 134° at the optimum sintering condition under nitrogen atmosphere. To the best of our knowledge, this is the first precisely study on wettability of Cu2O thin films prepared by this method at the various preparation conditions. [ABSTRACT FROM AUTHOR]

Arc-melting with ball milling was used for preparing Fe2CoGa1+x and Co2FeGa1+x (x=0,0.2) Heusler samples. Effect of extra nonmagnetic Ga atoms on lattice ordering and magnetic properties of Fe2CoGa and Co2FeGa Heusler compounds was studied. Rietveld refinement showed that lattice parameter of the samples increases in the presence of extra Ga atoms. Results showed that Co2FeGa1.2 has a partial lattice disordering. Saturation magnetization of Co2FeGa sample was lower than Slater -- Paulig prediction due to the crystallite size on a scale of nanometer. Reduction of saturation magnetization by increasing Ga atoms was explained through the change of lattice parameter, lattice ordering, and crystallite size. Saturation magnetization of Fe2CoGa sample was greater than Slater -- Paulig value. Change of lattice parameter by increasing Ga atoms resulted a decrease of saturation magnetization in Fe2CoGa1.2. [ABSTRACT FROM AUTHOR]

HRP, or horseradish peroxidase, is a reporter enzyme with extensive use in biotechnological applications. We previously reported the purification and characterization of two anionic peroxidases from Raphanus sativus L. var niger (black radish) roots. Here, we evaluated the applicability of these two novel peroxidases as alternatives to traditional horseradish peroxidase (HRP). The two novel peroxidases (BRP-A and BRP-B) and HRP were conjugated to IgY polyclonal antibodies by chemical methods based on the use of sodium periodate and cyanuric chloride. Moreover, the applicability of BRP-A and BRP-B in immunoassays was investigated by comparing the signal generated by these novel peroxidases in ELISA with HRP conjugates. Additionally, the limit of detection (LOD) was calculated for BRP-A, BRP-B, and HRP conjugates. Finally, the thermal stability of peroxidase antibody conjugates at 37 °C and 4 °C was compared. The peroxidase antibody conjugates prepared by the periodate method generated a much stronger signal than those prepared by the cyanuric chloride method. The signal obtained by BRP-A and BRP-B conjugates was much lower compared to the commercial HRP enzyme. The limit of detection was found to be 385.71, 213.75, and 43.6 ng per well for BRP-A, BRP-B, and HRP conjugates prepared by the periodate method, respectively. However, for conjugates prepared by the cyanuric chloride method, the limit of detection could only be estimated for HRP since BRP-A and BRP-B had an extremely low signal-to-noise ratio. All peroxidase conjugates had comparable thermal stability at 37 °C and 4 °C., Competing Interests: 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., (© 2024 The Authors.)

Social anxiety disorder is a prevalent mental health condition that significantly impairs social interactions, academic performance, and professional functioning in children, adolescents, and youth. This study aimed to investigate the global prevalence of social anxiety disorder across these developmental stages. Six electronic databases (PubMed, Scopus, Web of Science, Embase, ScienceDirect and Google Scholar) were systematically searched for studies related to the prevalence of social anxiety disorder in children, adolescents and youth. Random-effects models were employed for data analysis and Comprehensive Meta-Analysis Version 2.0. Heterogeneity was assessed using the I 2 index. A total of 38 studies were included in the final analysis. The global prevalence of social anxiety disorder was estimated to be 4.7% in children, 8.3% in adolescents, and 17% in youth. These findings suggest a progressive increase in the prevalence of SAD across these developmental stages. Considering the prevalence of social anxiety disorder in this study, policymakers can use the findings of this study to inform and develop effective prevention strategies for individuals and communities most susceptible to this disorder., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Rational design of synthetic phage-displayed libraries requires the identification of the most appropriate positions for randomization using defined amino acid sets to recapitulate the natural occurrence. The present study uses position-specific scoring matrixes (PSSMs) for identifying and randomizing Camelidae nanobody (VHH) CDR3. The functionality of a synthetic VHH repertoire designed by this method was tested for discovering new VHH binders to recombinant coagulation factor VII (rfVII)., Methods: Based on PSSM analysis, the CDR3 of cAbBCII10 VHH framework was identified, and a set of amino acids for the substitution of each PSSM-CDR3 position was defined. Using the Rosetta design SwiftLib tool, the final repertoire was back-translated to a degenerate nucleotide sequence. A synthetic phage-displayed library was constructed based on this repertoire and screened for anti-rfVII binders., Results: A synthetic phage-displayed VHH library with 1 × 10 8 variants was constructed. Three VHH binders to rfVII were isolated from this library with estimated dissociation constants (K D ) of 1 × 10 -8  M, 5.8 × 10 -8  M and 2.6 × 10 -  M.7  M., Conclusion: PSSM analysis is a simple and efficient way to design synthetic phage-displayed libraries., Competing Interests: Declaration of Competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Diphtheria can be prevented by vaccination, but some epidemics occur in several places, and diphtheria's threat is considerable. Administration of diphtheria antitoxin (DAT) produced from hyperimmunized animals is the most common treatment. Recombinant human antibody fragments such as single-chain variable fragments (scFv) produced by phage display library may introduce an interesting approach to overcome the limitations of the traditional antibody therapy. In the present study, B cells of immunized volunteers were used to construct a human single-chain fragment (HuscFv) library., Materials and Methods: The library was constructed with the maximum combination of heavy and light chains. As an antigen, Diphtheria toxoid (DTd) was used in four-round phage bio-panning to select phage clones that display DTd bound HuscFv from the library. After panning, individual scFv clones were selected. Clones that were able to detect DTd in an initial screening assay were transferred to Escherichia coli HB2151 to express the scFvs and purification was followed by Ni metal ion affinity chromatography. Toxin neutralization test was performed on Vero cells. The reactivity of the soluble scFv with diphtheria toxin were done and affinity calculation based on Beatty method was calculated., Results: The size of the constructed scFv library was calculated to be 1.3 × 10 6 members. Following four rounds of selection, 40 antibody clones were isolated which showed positive reactivity with DTd in an ELISA assay. Five clones were able to neutralize DTd in Vero cell assay. These neutralizing clones were used for soluble expression and purification of scFv fragments. Some of these soluble scFv fragments show neutralizing activity ranging from 0.6 to 1.2 µg against twofold cytotoxic dose of diphtheria toxin. The affinity constant of the selected scFv antibody was determined almost 10 7  M -1 ., Conclusion: This study describes the prosperous construction and isolation of scFv from the immune library, which specifically neutralizes diphtheria toxin. The HuscFv produced in this study can be a potential candidate to substitute the animal antibody for treating diphtheria and detecting toxins., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Staphylococcus aureus (S. aureus) is an opportunistic pathogen causing diseases ranging from mild skin infections to life threatening conditions, including endocarditis, pneumonia, and sepsis. To identify host genes modulating this host-pathogen interaction, we infected 25 Collaborative Cross (CC) mouse strains with methicillin-resistant S. aureus (MRSA) and monitored disease progression for seven days using a surgically implanted telemetry system. CC strains varied widely in their response to intravenous MRSA infection. We identified eight 'susceptible' CC strains with high bacterial load, tissue damage, and reduced survival. Among the surviving strains, six with minimal colonization were classified as 'resistant', while the remaining six tolerated higher organ colonization ('tolerant'). The kidney was the most heavily colonized organ, but liver, spleen and lung colonization were better correlated with reduced survival. Resistant strains had higher pre-infection circulating neutrophils and lower post-infection tissue damage compared to susceptible and tolerant strains. We identified four CC strains with sexual dimorphism: all females survived the study period while all males met our euthanasia criteria earlier. In these CC strains, males had more baseline circulating monocytes and red blood cells. We identified several CC strains that may be useful as new models for endocarditis, myocarditis, pneumonia, and resistance to MRSA infection. Quantitative Trait Locus (QTL) analysis identified two significant loci, on Chromosomes 18 and 3, involved in early susceptibility and late survival after infection. We prioritized Npc1 and Ifi44l genes as the strongest candidates influencing survival using variant analysis and mRNA expression data from kidneys within these intervals., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Nagarajan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: The diagnosis and treatment processes of cancer are among the main challenges of medical science in recent decades. The use of different therapeutic agents is one of the most common methods frequently utilized for cancer treatment. Accumulating evidence points to a potential effect of Obeticholic acid (OCA), a specific ligand for farnesoid X receptor, on the regulation of cancer-associated pathways. In spite of tremendous efforts to introduce OCA into the clinical setting, there is a great deal of uncertainty about its impact on breast cancer treatment. This study was performed to evaluate the effects of OCA on breast cancer., Methods and Results: In this experiment, the MCF-7 (Michigan Cancer Foundation-7) cell line was treated with 0.1 µM OCA, and cancerous characteristics of the MCF-7 cell line was evaluated by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2 H-tetrazolium bromide) assay, gelatin zymography, western blot, Real-time PCR, flow cytometry, and ELISA techniques. The results indicated that OCA increased the rate of apoptosis and the expression levels of PPARα (Peroxisome proliferator-activated receptor alpha) and TIMP-1 (tissue inhibitor of metalloproteinase-1) genes in this cell line, while it reduced the mRNA levels of MMP7 (matrix metalloproteinase 7) and Bcl-2 (B-cell lymphoma 2) genes, as well as the protein levels of the active form of AKT (protein kinase B), Erk1/2 (extracellular signal-regulated kinase 1/2) and STAT3 (Signal transducers and activators of transcription-3). Also, OCA decreased the activity of MMP9, while it increased the secretion of VEGF-A (vascular endothelial growth factor-A)., Conclusions: It seems that OCA can exert anti-cancer effects on the MCF-7 cells by reducing growth, proliferation, migration, invasion, and regulation of the expression of genes involved in cancer-associated pathways. However, it should be noted that further studies are warranted to establish this concept, especially the increase of VEGF-A can be considered a challenge for the results of this study., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Cholesterol oxidase is industrially important as it is frequently used as a biosensor in food and agriculture industries and measurement of cholesterol. Although, most natural enzymes show low thermostability, which limits their application. Here, we obtained an improved variant of Chromobacterium sp. DS1 cholesterol oxidase (ChOS) with enhanced thermostability by random mutant library applying two forms of error-prone PCR (serial dilution and single step). Wild-type ChOS indicated an optimal temperature and pH of 70 ºC and pH 7.5, respectively. The best mutant ChOS-M acquired three amino acid substitutions (S112T, I240V and A500S) and enhanced thermostability (at 50 °C for 5 h) by 30%. The optimum temperature and pH in the mutant were not changed. In comparison to wild type, circular dichroism disclosed no significant secondary structural alterations in mutants. These findings show that error-prone PCR is an effective method for enhancing enzyme characteristics and offers a platform for the practical use of ChOS as a thermal-resistance enzyme in industrial fields and clinical diagnosis., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Tetanus is an acute and often fatal infectious disease caused by Clostridium tetani . Tetanus toxin (TT) is responsible for spastic paralysis observed in tetanus. Anti-tetanus antibodies obtained from horses and humans are the most antitoxins used for tetanus treatment, although some clinical side effects and disadvantages have been reported in their application. The aim of this study is the production of anti-TT IgY and evaluation of its protective effects in a mouse model. Anti-TT IgY was purified from the egg yolk using PEG6000 precipitation and water dilution methods, and its purity was verified by SDS-PAGE. Finally, the potency of purified anti-TT IgY in neutralizing the lethal effects of TT was studied in vivo using a mouse model. PEG6000 precipitation method had better results. Animal studies showed that the purified IgY neutralized the toxic effects of 100 MLD of TT and multiple intravenous-dose injections of anti-TT IgY also had a continuous effect of TT neutralization. The purified anti-TT IgY was effective in neutralizing the lethal activity of TT in a mouse model. Our results suggested that IgY could be an alternative therapeutic source for the management of tetanus in the future. Abbreviations Anti-TT, Anti-tetanus toxin; ELISA, Enzyme-linked immunosorbent assay; IgY, Immunoglobulin Y; MLD, Minimum lethal dose; PBS, Phosphate buffer solution; PEG, Polyethylene glycol; SDS-PAGE, Sodium dodecyl sulfate polyacrylamide gel electrophoresis; TIG, Tetanus immune globulin; TT, Tetanus toxin; WD, Water dilution; RT, Room temperature.

Background: Thimerosal (Merthiolate) is a well-known preservative used in pharmaceutical products, the safety of which was a matter of controversy for decades. Thimerosal is a mercury compound, and there is a debate as to whether Thimerosal exposure from vaccination can contribute to the incidence of mercury-driven disorders. To date, there is no consensus on Thimerosal safety in Vaccines. In 1977, a maximum safe dose of 200 μg/ml (0.5 mM) was recommended for Thimerosal by the WHO experts committee on biological standardization. Up-to-date guidelines, however, urge national control authorities to establish their own standards for the concentration of vaccine preservatives. We believe such safety limits must be studied at the cellular level first. The present study seeks a safe yet efficient dose of Thimerosal exposure for human and animal cells and control microorganism strains., Methods: The safety of Thimerosal exposure on cells was analyzed through an MTT cell toxicity assay. The viability of four cell types, including HepG2, C2C12, Vero Cells, and Peripheral blood mononuclear cells (PBMCs), was examined in the presence of different Thimerosal concentrations and the maximum tolerable dose (MTD) and the half maximal inhibitory concentration (IC50) values for each cell line were determined. The antimicrobial effectiveness of Thimerosal was evaluated on four control strains, including Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and Aspergillus brasiliensis, to obtain the minimum inhibitory concentration (MIC) of Thimerosal. The MIC test was performed in culture media and under optimal growth conditions of microorganisms in the presence of different Thimerosal concentrations., Results: The viability of all examined cell lines was suppressed entirely in the presence of 4.6 μg/ml (12.5 μM) of Thimerosal. The MTD for HepG2, C2C12, PBMC, and Vero cells was 2, 1.6, 1, and 0.29 μg/ml (5.5, 4.3, 2.7 and 0.8 μM), respectively. The IC50 of Thimerosal exposure for HepG2, C2C12, PBMC, and Vero cells was 2.62, 3.17, 1.27, and 0.86 μg/ml (7.1, 8.5, 3.5 and 2.4 μM), respectively. As for antimicrobial effectiveness, the growth capability of Candida albicans and Staphylococcus aureus was suppressed entirely in the presence of 6.25 µg/ml (17 μM) Thimerosal. The complete growth inhibition of Pseudomonas aeruginosa in culture media was achieved in 100 µg/ml (250 µM) Thimerosal concentration. This value was 12.5 µg/ml (30 μM) for Aspergillus brasiliensis., Conclusion: According to our results Thimerosal should be present in culture media at 100 μg/ml (250 µM) concentration to achieve an effective antimicrobial activity. We showed that this amount of Thimerosal is toxic for human and animal cells in vitro since the viability of all examined cell lines was suppressed in the presence of less than 5 μg/ml (12.5 μM) of Thimerosal. Overall, our study revealed Thimerosal was 333-fold more cytotoxic to human and animal cells as compared to bacterial and fungal cells. Our results promote more study on Thimerosal toxicity and its antimicrobial effectiveness to obtain more safe concentrations in biopharmaceuticals., 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 © 2023 Elsevier GmbH. All rights reserved.)

Given an infected host, estimating the time that has elapsed since initial exposure to the pathogen is an important problem in public health. In this paper we use longitudinal gene expression data from human challenge studies of viral respiratory illnesses for building predictive models to estimate the time elapsed since onset of respiratory infection. We apply sparsity driven machine learning to this time-stamped gene expression data to model the time of exposure by a pathogen and subsequent infection accompanied by the onset of the host immune response. These predictive models exploit the fact that the host gene expression profile evolves in time and its characteristic temporal signature can be effectively modeled using a small number of features. Predicting the time of exposure to infection to be in first 48 h after exposure produces BSR in the range of 80-90% on sequestered test data. A variety of machine learning experiments provide evidence that models developed on one virus can be used to predict exposure time for other viruses, e.g., H1N1, H3N2, and HRV. The interferon [Formula: see text] signaling pathway appears to play a central role in keeping time from onset of infection. Successful prediction of the time of exposure to a pathogen has potential ramifications for patient treatment and contact tracing., (© 2023. The Author(s).)

Understanding the molecular mechanisms underlying resistance and tolerance to pathogen infection may present the opportunity to develop novel interventions. Resistance is the absence of clinical disease with a low pathogen burden, while tolerance is minimal clinical disease with a high pathogen burden. Salmonella is a worldwide health concern. We studied 18 strains of collaborative cross mice that survive acute Salmonella Typhimurium (STm) infections. We infected these strains orally and monitored them for 3 weeks. Five strains cleared STm (resistant), six strains maintained a bacterial load and survived (tolerant), while seven strains survived >7 days but succumbed to infection within the study period and were called "delayed susceptible." Tolerant strains were colonized in the Peyer's patches, mesenteric lymph node, spleen, and liver, while resistant strains had significantly reduced bacterial colonization. Tolerant strains had lower preinfection core body temperatures and had disrupted circadian patterns of body temperature postinfection sooner than other strains. Tolerant strains had higher circulating total white blood cells than resistant strains, driven by increased numbers of neutrophils. Tolerant strains had more severe tissue damage and higher circulating levels of monocyte chemoattractant protein 1 (MCP-1) and interferon gamma (IFN-γ), but lower levels of epithelial neutrophil-activating protein 78 (ENA-78) than resistant strains. Quantitative trait locus (QTL) analysis revealed one significant association and six suggestive associations. Gene expression analysis identified 22 genes that are differentially regulated in tolerant versus resistant animals that overlapped these QTLs. Fibrinogen genes ( Fga , Fgb , and Fgg ) were found across the QTL, RNA, and top canonical pathways, making them the best candidate genes for differentiating tolerance and resistance. IMPORTANCE To survive a bacterial infection, an infected host can display resistance or tolerance. Resistance is indicated by a decrease in pathogen load, while for tolerance a high pathogen load is accompanied by minimal disease. We infected genetically diverse mice with Salmonella Typhimurium for 21 days and discovered new phenotypes for disease outcome (delayed susceptible, tolerant, and resistant). Tolerant strains showed the lowest preinfection core body temperatures and the most rapid disruption in circadian patterns of body temperature postinfection. Tolerant strains had higher circulating neutrophils and higher circulating levels of MCP-1 and IFN-γ, but lower levels of ENA-78 than did resistant strains, in addition to more severe tissue damage. QTL analysis revealed multiple associated regions, and gene expression analysis identified 22 genes that are differentially regulated in tolerant versus resistant animals in these regions. Fibrinogen genes ( Fga , Fgb , and Fgg ) were found across the QTL, RNA, and the top canonical pathways, suggesting a role in tolerance.

Peroxidase is a commonly used enzyme with a wide range of applications. Horseradish (Armoracia rusticana) is the most well-known source of peroxidase enzyme. Peroxidases extracted from other plant sources have also been proved as useful, sometimes even superior, comparing to traditional horseradish peroxidase (HRP). In the present study, two novel peroxidase isoenzymes were purified and characterized from Raphanus sativus L. var niger roots. Two anionic peroxidase isoenzymes were purified using diafiltration, ammonium sulfate precipitation, DEAE anion-exchange chromatography, and concanavalin A affinity chromatography. The heaviest anionic isoenzyme (isoenzyme A) has a MW of about 110 KD, and the other anionic isoenzyme (isoenzyme B) has a MW of 97 KD. Both isoenzymes have an optimum temperature of 40 °C, but the activity of isoenzyme B is much more dependent on temperature with a Q10 of 3.5, while isoenzyme A has a Q10 of 1.7. These isoenzymes showed great thermal stability at 37 °C and 4 °C. Isoenzyme A showed the highest activity at pH 5 and it was found to be more stable at pH 6, whereas isoenzyme B showed the highest activity at pH 6 and is more stable at pH 7. Isoenzyme A has a K m value of 10.63 mM and 0.043 mM, and isoenzyme B has a K m of 15.38 mM and 0.067 mM for 4-aminoantipyrine and H 2 O 2 , respectively. The isoenzymes purified from Raphanus sativus L. var niger offer excellent chemical and thermal stability, which encourages further studies on their suitability for biotechnological applications., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Salmonella infections typically cause self-limiting gastroenteritis, but in some individuals these bacteria can spread systemically and cause disseminated disease. Salmonella Typhimurium (STm), which causes severe systemic disease in most inbred mice, has been used as a model for disseminated disease. To screen for new infection phenotypes across a range of host genetics, we orally infected 32 Collaborative Cross (CC) mouse strains with STm and monitored their disease progression for seven days by telemetry. Our data revealed a broad range of phenotypes across CC strains in many parameters including survival, bacterial colonization, tissue damage, complete blood counts (CBC), and serum cytokines. Eighteen CC strains survived to day 7, while fourteen susceptible strains succumbed to infection before day 7. Several CC strains had sex differences in survival and colonization. Surviving strains had lower pre-infection baseline temperatures and were less active during their daily active period. Core body temperature disruptions were detected earlier after STm infection than activity disruptions, making temperature a better detector of illness. All CC strains had STm in spleen and liver, but susceptible strains were more highly colonized. Tissue damage was weakly negatively correlated to survival. We identified loci associated with survival on Chromosomes (Chr) 1, 2, 4, 7. Polymorphisms in Ncf2 and Slc11a1, known to reduce survival in mice after STm infections, are located in the Chr 1 interval, and the Chr 7 association overlaps with a previously identified QTL peak called Ses2. We identified two new genetic regions on Chr 2 and 4 associated with susceptibility to STm infection. Our data reveal the diversity of responses to STm infection across a range of host genetics and identified new candidate regions for survival of STm infection., Competing Interests: The authors have declared that no competing interests exist.

To enhance the thermal stability of Streptomyces Sp. SA-COO cholesterol oxidase, random mutagenesis was used. A random mutant library was generated using two types of error-prone PCR (single step and serial dilution) and two mutants (ChOA-M1 and ChOA-M2) with improved thermostability were obtained. The best mutant ChOA-M1 acquired three amino acid substitutions (G49T, W52K, and F62V) and improved thermostability (at 50 °C for 5 h) by 40% and increased the kcat/Km value by 23%. The optimum pH was desirably changed to encompass a broad range from alkali to acid and circular dichroism revealed no significant secondary structure changes in mutants against wild type. These findings indicated that random mutagenesis was an effective technique for optimizing cholesterol oxidase properties and make a foundation for practical applications of Cholesterol oxidase in clinical diagnosis and industrial fields., (Copyright © 2021. Published by Elsevier Inc.)

The COVID-19 pandemic crisis has fundamentally changed the way we live and work forever. The business sector is forecasting and formulating different scenarios associated with the impact of the pandemic on its employees, customers, and suppliers. Various business retrieval models are under construction to cope with life after the COVID-19 Pandemic Crisis. However, the proposed plans and scenarios are static and cannot address the dynamic pandemic changes worldwide. They also have not considered the peripheral in-between scenarios to propel the shifting paradigm of businesses from the existing condition to the new one. Furthermore, the scenario drivers in the current studies are generally centered on the economic aspects of the pandemic with little attention to the social facets. This study aims to fill this gap by proposing scenario planning and analytics to study the impact of the Coronavirus pandemic on large-scale information technology-led Companies. The primary and peripheral scenarios are constructed based on a balanced set of business continuity and employee health drivers. Practical action plans are formulated for each scenario to devise plausible responses. Finally, a damage management framework is developed to cope with the mental disorders of the employees amid the disease., Competing Interests: 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., (© 2021 The Author(s).)

Coronavirus disease 2019 (COVID-19) is a respiratory disease that outbreaks since December 2019 and spread globally. Various methods have been used to treat SARS-CoV-2 that is generally based on the information obtained from the therapeutic approaches used for SARS-COV and MERS patients. In this article, we introduce a theoretical strategy in which a two-domain fusion protein presents the virus to the immune system. This fusion protein contains a viral-binding domain such as the ACE2 domain and a domain such as the hepatitis B antigen that has previously been exposed to the immune system. This two-domain fusion protein, could be called "virus-presenting fusion protein", would attach to the virus spike protein via the ACE2 domain while the hepatitis B antigen would be bound by anti-hepatitis B antibodies facilitating the opsonization and presentation of the virus to the immune system. We believe that this virus-presenting fusion protein will accelerate the immune response to the SARS-CoV-2 virus., (© Indian Virological Society 2021.)

The clinical importance of blood group (BG) antigens is related to their ability to induce immune antibodies that can cause hemolysis. Yet, ABO and D (Rh) are still considered to be the key antigens for healthy blood transfusion and secondary antigens are the next priority. Serological typing is the most widely used typing method. Rapid and accurate blood grouping plays an important role in some clinical conditions, rather than conventional techniques. Hence, developing a simple and economical model for rapid blood grouping would facilitate these tests. In recent decades, paper-based microfluidics such as μPADs has gained much interest in wide application areas such as point-of-care diagnostic. In this study, we evaluated μPADs that are performed for blood grouping and its recent progress. A comprehensive literature search was performed using databases including PUBMED, SCOPUS, Web of Science and Google Scholar. Keywords were blood grouping or typing, paper analytical device, rapid test, etc. After investigation of search results, 16 papers from 2010 to 2020 were included. Further information in detail was classified in Table 1. Generally, two principles for blood typing μPADs are introduced. The lateral chromatographic flow method and the vertical flow-through method that detects BG in a visual-based manner. To detect results with acceptable clarity many factors and challenges like paper, blood sample, buffer, Ab and RBC interaction and also μPADs stability need to be considered, which are discussed. In conclusion, the simplicity, stability, cheapness, portability and biocompatibility of μPADs for blood grouping confirming its utility and also they have the capability to robust, universal blood-grouping platform. Table 1 Summary of blood grouping tests using paper-based analytical devices Antigens Type of diagnosis Validation method Sample No Accuracy Action time Paper type Stability Sample dilution Buffer Ref A, B, Rh Forward volunteers records 5 - - Whatman No. 4 - 1/2 PBS* (Khan et al. 2010) A, B, Rh Forward gel assay test and conventional slide test 100 100% 1 min Whatman No. 4 and Kleeenex paper towel 7 Days in 4 °C 1/1 NSS (Al-Tamimi et al. 2012) A, B, Rh Forward gel card assay 99 100% 20 Sec + Washing Kleeenex paper towel - 1/1 NSS (Li et al. 2012) A, B, Rh Forward - - - - Kleeenex paper towel - 45/100 PSS (Li et al. 2013) A, B, Rh Forward gel card assay 98 100% 1.5 min Kleeenex paper towel - 85/100 PBS (Guan et al. 2014b) C, E, c, e, K, Jka, Jkb, M, N, S, P1, and Lea Forward gel card assay 266 100% - Kleeenex paper towel - 1/1 NSS (Li et al. 2014b) A, B, Rh Forward and Reverse conventional slide test 96 ≈ 91% 10 min Whatman No. 1 21 Days in 4 °C 1/2 NSS (Noiphung et al. 2015) C, c, E, e, K, k, Fya, Fyb, Jka, Jkb, M, N, S and s, P1, Lea and Leb Forward - 478 - - Kleeenex paper towel - 1/1 NSS, PBS (Then et al. 2015) A, B Forward and Reverse conventional slide test 76 100% 5-8 min Whatman No. 4 38 Days in 4 °C 1/4, 1/1 NSS (Songjaroen and Laiwattanapaisal 2016) D, K Forward volunteers records 210 - 7.5 min Kleenex paper towel - 1/1 NSS (Yeow et al. 2016) A, B, c, e, D, C, E, M, N, S, s, P1, Jka, Jkb, Lea, Leb, Fya, and Fyb Forward and Reverse gel card assay 3550 ≈100% 30 s Fiber glass and cotton linter 180 Days in 25 °C 45/100, 1/1 PBS (Zhang et al. 2017) A, B Forward conventional slide test 598 100% 3 min Whatman No. 113 14 Day in 4 °C 1/1 NSS (Songjaroen et al. 2018) A, B, Rh Forward conventional slide test - - 30 Sec + Washing Unrefined sisal paper - 1/2 NSS (Casals-Terré et al. 2019) A, B, Rh Forward - - - - Whatman No.1 - 1/1 NSS (Ansari et al. 2020) ABO & Rh Forward and Reverse conventional slide test - 100% Unrefined Eucalyptus papers - 1/2 NSS, PBS (Casals-Terré et al. 2020) A, B, Rh Forward - - - 30 Sec + Washing Whatman No. 4 modified with chitosan  ≥ 100 days in 25 °C 1/1 NSS (Parween et al. 2020) * phosphate buffer saline, normal saline solution.

Background: Diphtheria is a bacterial disease which is caused by Corynebacterium diphtheriae. The symptoms are due to the diphtheria toxin produced by the bacteria. Antibiotic therapy and the use of diphtheria antitoxin is a recommended strategy to control diphtheria. Although mammalian antibodies are used to treat patients, IgY antibody has advantages over mammalian ones, including cost-effectiveness and production through non-invasive means. Moreover, in contrast to mammalian antibodies, IgY does not bind to the rheumatoid factor and does not activate the complement system. The objective of this study was to evaluate the in vitro neutralizing effect of IgY against diphtheria toxin., Results: Anti-DT IgY was produced by immunization of the laying white leghorn chickens. Indirect enzyme-linked immunosorbent assay revealed successful immunization of the animals, and the IgY was purified with a purity of 93% via polyethylene glycol precipitation method. The neutralizing activity of the purified IgY was evaluated by Vero cell viability assay. This assay confirmed that 1.95 μg (8.6 μg/ml of culture medium) of anti-DT IgY would neutralize 10 fold of cytotoxic dose 99% of DT, which was 0.3 ng (1.33 ng/ml of culture medium)., Conclusion: This anti-DT IgY may be applicable for diphtheria treatment and quality controls in vaccine production.

Background: Antimicrobial photodynamic therapy (aPDT) as a complementary step of selective removal of dental caries appears as an ideal choice to help manage caries with minimal interventions. Sub-lethal dose of aPDT (sPDT) impressively effects microbial virulence, although it does not kill microorganisms. Therefore, the present study aimed to investigate the influence of sPDT using diode laser plus chlorophyllin-phycocyanin mixture (PhotoActive + ) on changes in gtfB gene expression and the subsequent protein expression of GtfB., Materials and Methods: sPDT using PhotoActive + and 635 ± 10 nm diode laser was used to examine the expression of gtfB mRNA and GtfB protein expression of Streptococcus mutans by quantitative real time PCR (qRT-PCR) method and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), respectively., Results: In this study, sPDT using 2.4 × 10 -3 mol/L PhotoActive + with 3 min irradiation time of diode laser with energy density of 104 J/cm 2 , significantly reduced the gtfB gene expression with rates of 3.5-fold (P < 0.05). Also, PhotoActive + mediated sPDT demonstrated a significant reduction in GtfB protein expression of S. mutans up to 54 % (P < 0.05)., Conclusions: This study demonstrated the potential effect of PhotoActive + mediated sPDT on one of the most important virulence factors of S. mutans., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objectives: Melatonin induces human stem cells, converts pre-osteoblasts to mature osteoblasts, and reduces the duration of this transition. However, melatonin itself prevents activation of osteoclasts. Here, we evaluate the role of melatonin in prevention of bisphosphonate-related osteonecrosis of the jaw., Materials and Methods: In this experimental-interventional study, 30 rats were evaluated in 3 groups. The first and second groups received saline and zoledronic acid, respectively, for 4 weeks and the third group received 4 weeks of zoledronic acid and 3 weeks of melatonin simultaneously. Firstright- maxillary-molar extraction was performed for all animals, which were sacrificed after 4 weeks of recovery. The extraction sockets were examined histologically for the presence of osteonecrosis, number of osteoclasts and fibroblasts, severity of inflammation, and vascularization. Data were analyzed by chi-square, one-way ANOVA, Tukey, Kruskal-Wallis and Fisher's exact statistical tests (α=0.05)., Results: Osteonecrosis was observed in 20%, 90%, and 70% of the first, second and third groups, respectively ( P =0.008). The lowest number of osteoclasts and fibroblasts was seen in the third group., Conclusion: Melatonin may effectively prevent some undesirable side effects of bisphosphonates. However, further studies are required to confirm the results of this study.

Background: Two-dimensional intraoral radiography is the most common tool for recognizing root fractures. Improving the quality of images by means of enhancement tools can increase the recognition power of them. The aim of this study is to evaluate the effect of digital image enhancement on vertical and horizontal root fractures (HRFs) diagnostic accuracy., Materials and Methods: In this in vitro study, 100 human extracted teeth, involving 50 mandibular premolars and 50 maxillary incisors, were investigated. In total, 25 premolar teeth were vertically fractured and other 25 sound teeth served as testing group. According to the verified methods, 25 incisor teeth were fractured and other 25 teeth of this group served as testing ones. Following, by using the charge-coupled device sensor, preapical digital images were recorded. The original images were altered using reverse-contrast and colorization enhancement tools. Two different observers independently investigated all of the images. Receiver operating characteristic analysis was used to calculate the area under the curve (AUC) and sensitivity and specificity of all images. Data analyzde using receiver operating characteristic (ROC) analysis. Two-ways variance analysis was used to assess differences in the values ( P = 0.05 )., Results: AUC and sensitivity and specificity related to the original, reverse-contrast, and colorized images were calculated (0.84, 0.64, 0.99), (0.84, 0.64, 0.96), and (0.82, 0.64, 0.92) respectively, for vertically root fractured images. AUC and sensitivity and specificity related to the original, reverse-contrast, and colorized images were calculated (0.49, 0.44, 0.56), (0.50, 0.44, 0.60), and (0.48, 0.48, 0.48), respectively, for horizontally root-fractured images., Conclusion: The results of the present study revealed that reverse-contrast and colorized enhancement filters cannot be used as critical methods in detecting in vitro vertical and HRF., Competing Interests: The authors of this manuscript declare that they have no conflicts of interest, real or perceived, financial or nonfinancial in this article., (Copyright: © 2020 Dental Research Journal.)

Background: Partial (selective) removal of dental caries is a suitable manner to treat deep carious lesions in vital teeth with asymptomatic pulps. Antimicrobial photodynamic therapy (aPDT) was proposed as a promising ancillary approach for reduction of the residual bacteria from the cavity. Therefore, the focus of this study was to investigate the influence of aPDT using diode laser (DL) plus PhotoActive+ (chlorophyllin-phycocyanin mixture [CHL-PC]) as photosensitizer (PS) on metabolic activity and the reduction in the number of living bacteria within the preformed biofilm caries model on enamel slabs of Streptococcus mutans., Materials and Methods: The lethal and sub-significant inhibitory (SSI) potential of aPDT using CHL-PC and 635 nm DL against experimental biofilm caries model on enamel slabs and metabolic activity of S. mutans was analyzed using crystal violet and XTT reduction assays, respectively. Intracellular ROS formation by DCFH-DA assay was measured in CHL-PC mediated aPDT treated bacterial samples. Tooth discoloration and cell cytotoxicity of CHL-PC were assessed in the CIEL*a*b* color space and neutral red assay, respectively., Results: In this study aPDT at a maximum concentration level of CHL-PC (5000 μg/mL) with 3 min DL irradiation time (103.12 J/cm 2 ) reduced the ex-vivo cariogenic biofilm of S. mutans by 36.93 % (P <  0.05). Although chlorhexidine (CHX) had an anti-biofilm effect about 1.7 fold compared to CHL-PC mediated aPDT, this difference was not significant (36.93 in comparison to 63.05 %; P >  0.05). CHL-PC mediated aPDT demonstrated a significant reduction in bacterial metabolic activity, with rates of 77 % at a SSI dose (using 156 μg/mL of CHL-PC and 3 min DL irradiation time with the energy density of 103.12 J/cm 2 ). The treated bacterial cells exhibited significant (P < 0.05) increment in the ROS generation. The least color change (ΔE) was found using CHL-PC at a concentration of 156 μg/mL (ΔE = 2.74). CHL-PC in different concentrations showed no significant reduction in human gingival fibroblasts (HGFs) cell survival (P >  0.05)., Conclusion: CHL-PC mediated aPDT not only reduces the number of living bacteria within the biofilms of S. mutans in an experimental biofilm caries model on enamel slabs but also its influences microbial virulence by reducing the metabolic activity of the S. mutants., (Copyright © 2019. Published by Elsevier B.V.)

Recent developments in both biological data acquisition and analysis provide new opportunities for data-driven modelling of the health state of an organism. In this paper, we explore the evolution of temperature patterns generated by telemetry data collected from healthy and infected mice. We investigate several techniques to visualize and identify anomalies in temperature time series as temperature relates to the onset of infectious disease. Visualization tools such as Laplacian Eigenmaps and Multidimensional Scaling allow one to gain an understanding of a dataset as a whole. Anomaly detection tools for nonlinear time series modelling, such as Radial Basis Functions and Multivariate State Estimation Technique, allow one to build models representing a healthy state in individuals. We illustrate these methods on an experimental dataset of 306 Collaborative Cross mice challenged with Salmonella typhimurium and show how interruption in circadian patterns and severity of infection can be revealed directly from these time series within 3 days of the infection event., Competing Interests: We declare that we have no competing interests., (© 2019 The Author(s).)

The aggregation behavior of particles in nonpolar media is studied with time-resolved light scattering. At low surfactant concentrations particles are weakly charged and suspensions are not stable. The suspensions become progressively more stable with increasing surfactant concentration as particles become more highly charged. At high concentrations the particles become neutralized and aggregation is again fast. The theory of Derjaguin, Landau, Verwey, and Overbeek (DLVO) is able to predict the stability ratios quantitatively by using the experimentally measured surface charges, screening lengths and van der Waals forces.

The biopanning process is a critical step in phage display for isolating peptides or proteins with specific binding properties. Conventional panning methods are sometimes not so effective and may result in nonspecific or low-yield positive results. In this study, three different strategies including soluble antibody-capturing, pH-stepwise elution, and conventional panning were used for enrichment of specific clones against diphtheria toxoid. The reactivity of the selected clones was evaluated using an indirect enzyme-linked immunosorbent assay. The positive clones were screened using Vero cell viability assay. The neutralizing clones were expressed in HB2151 strain of Escherichia coli and soluble single-chain fragment variable (scFv) fragments were purified by nickel-nitrilotriacetic acid affinity chromatography. Finally, the ability of scFv fragments for neutralizing diphtheria toxin (DT) were evaluated again using Vero cell viability assay. After four rounds of panning, the soluble antibody-capturing method yielded 15 positive phage-scFv clones against diphtheria toxoid. Conventional panning and pH-stepwise elution model resulted from nine and five positive phage-scFv clones, respectively. Among all positive clones, three clones were able to neutralize DT in Vero cell viability assay. Two of these clones belonged to a soluble antibody-capturing method and one of them came from conventional panning. Three neutralizing clones were used for soluble expression and purification of scFvs fragments. It was found that these soluble scFv fragments possessed neutralizing activity ranging from 0.15 to 0.6 µg against two-fold cytotoxic dose 99% of DT. In conclusion, the results of our study indicate that soluble antibody-capturing method is an efficient method for isolation of specific scFv fragments., (© 2018 Wiley Periodicals, Inc.)

Immune response stimulation and inactivation of chondroitinase ABC I in physiological condition have been limited its use in various clinical conditions as a bacterial enzyme drug. In the present study, we have investigated some structural and functional features of N∆89, C∆274 and N∆89C∆274; three designed truncated cABC I, in order to clarify the unclear role of two terminal parts of cABC I i.e., the 1-89 and 747-1021 amino acids sequences of the full length enzyme through truncation. As a result, the numbers of potential epitopes, the susceptibility to trypsin digestion, ANS fluorescence spectra, and fluorescence quenching using KI and acrylamide were diminished for N∆89 and C∆274 in comparison to the wild type. Secondary and tertiary structure investigation for N∆89 and C∆274 revealed that the intrinsic fluorescence was increased and Far-UV CD spectra were changed accordingly. Relative to the wild type enzyme, 0.164, 0.195 remaining activity and lack of activity was shown with the zymographic assay for N∆89, C∆274 and N∆89C∆274 variants, respectively. The diminished enzyme activity and structural changes suggested a reorientation of microenvironments interactions including cation-π interactions around structural elements toward lowering regional mobility. Constructing applicable truncated cABC I with improved features could be regarded as a strategy to regain new possible functional advantages over the full length enzyme.

Cholesterol oxidase is a bifunctional bacterial flavoenzyme which catalyzes oxidation and isomerization of cholesterol. This valuable enzyme has attracted a great deal of attention because of its wide application in the clinical laboratory, synthesis of steroid derived drugs, food industries, and its potentially insecticidal activity. Therefore, development of an efficient protocol for overproduction of cholesterol oxidase could be valuable and beneficial in this regard. The present study examined the role of various parameters (host strain, culture media, induction time, isopropyl ß-D-1-thiogalactopyranoside concentration, as well as post-induction incubation time and temperature) on over-expression of cholesterol oxidase from Chromobacterium sp. DS1. Applying the optimized protocol, the yield of recombinant cholesterol oxidase significantly increased from 92 U/L to 2115 U/L. Under the optimized conditions, the enzyme was produced on a large-scale, and overexpressed cholesterol oxidase was purified from cell lysate by column nickel affinity chromatography. Km and Vmax values of the purified enzyme for cholesterol were estimated using Lineweaver-Burk plot. Further, the optimum pH and optimum temperature for the enzyme activity were determined. This study reports a straightforward protocol for cholesterol oxidase production which can be performed in any laboratory., Competing Interests: The authors have declared that no competing interests exist.

Cholesterol oxidase is a bacterial flavoenzyme which catalyzes oxidation and isomerization of cholesterol. This enzyme has a great commercial value because of its wide applications in cholesterol analysis of clinical samples, synthesis of steroid-derived drugs, food industries, and potentially insecticidal activity. Accordingly, development of an efficient protocol for overexpression of cholesterol oxidase can be very valuable and beneficial. In this study, expression optimization of cholesterol oxidase from Streptomyces sp. SA-COO was investigated in Escherichia coli host strains. Various parameters that may influence the yield of a recombinant enzyme were evaluated individually. The optimal host strain, culture media, induction time, Isopropyl ß-D-1-thiogalactopyranoside concentration, as well as post-induction incubation time and temperature were determined in a shaking flask mode. Applying the optimized protocol, the production of recombinant cholesterol oxidase was significantly enhanced from 3.2 to 158 U/L. Under the optimized condition, the enzyme was produced on a large-scale, and highly expressed cholesterol oxidase was purified from cell lysate by column nickel affinity chromatography. K m and V max values of the purified enzyme for cholesterol were estimated using Lineweaver-Burk plot. Further, the optimum pH and optimum temperature for the enzyme activity were also determined. We report a straightforward and easy protocol for cholesterol oxidase production which can be performed in any laboratory.

High yield recombinant protein production is highly desirable for biotechnological purposes. In the design of recombinant expression conditions, a number of essential central elements such as expression strain, type of medium, bioprocess optimization, and mathematical modeling should be considered. Well-designed industrial scale production of one recombinant protein with optimized influential parameters and yield can address the cost and production reproducibility issues. In the present study, statistical experimental design methodology was used to investigate the effect of fermentation conditions (dissolved oxygen, IPTG, and temperature) on rPDT production by Escherichia coli. rPDT is a recombinant fusion protein consisting of three different protein domains including the N-terminal 179 amino acid fragment of the S1 subunit of pertussis toxin, the full-length genetically detoxified diphtheria toxin (CRM197), and the 50 kDa tetanus toxin fragment C. A 15 Box-Behnken design augmented with center points revealed that IPTG and DO at the center point and low temperature will result in high yield. The optimal condition for rPDT production were found to be 100 µM IPTG, DO 30% and temperature 20 °C.

A simple method to experimentally observe and measure the dispersion of a passive tracer in a laminar fluid flow is described. The method consists of first injecting fluorescent dye directly into a pipe filled with distilled water and allowing it to diffuse across the cross-section of the pipe to obtain a uniformly distributed initial condition. Following this period, the laminar flow is activated with a programmable syringe pump to observe the competition of advection and diffusion of the tracer through the pipe. Asymmetries in the tracer distribution are studied and correlations between the pipe cross-section and the shape of the distribution is shown: thin channels (aspect ratio << 1) produce tracers arriving with sharp fronts and tapering tails (front-loaded distributions), while thick channels (aspect ratio ~1) present the opposite behavior (back-loaded distributions). The experimental procedure is applied to capillary tubes of various geometries and is particularly relevant to microfluidic applications by dynamical similarity.

Objectives: Foreign bodies (FBs) account for 3.8% of all pathologies of the head and neck region, and approximately one third of them are missed on initial examination. Thus, FBs represent diagnostic challenges to maxillofacial surgeons, rendering it necessary to employ an appropriate imaging modality in suspected cases., Materials and Methods: In this cross-sectional study, five different materials, including wood, metal, glass, tooth and stone, were prepared in three sizes (0.5, 1, and 2 mm) and placed in three locations (soft tissue, air-filled space and bone surface) within a sheep's head (one day after death) and scanned by panoramic radiography, cone-beam computed tomography (CBCT), and ultrasonography (US) devices. The images were reviewed, and accuracy of the detection modalities was recorded. The data were analyzed statistically using the Kruskal-Wallis, Mann-Whitney U-test, Friedman, Wilcoxon signed-rank and kappa tests ( P <0.05)., Results: CBCT was more accurate in detection of FBs than panoramic radiography and US ( P <0.001). Metal was the most visible FB in all of modalities. US was the most accurate technique for detecting wooden materials, and CBCT was the best modality for detecting all other materials, regardless of size or location ( P <0.05). The detection accuracy of US was greater in soft tissue, while both CBCT and panoramic radiography had minimal accuracy in detection of FBs in soft tissue., Conclusion: CBCT was the most accurate detection modality for all the sizes, locations and compositions of FBs, except for the wooden materials. Therefore, we recommend CBCT as the gold standard of imaging for detecting FBs in the maxillofacial region., Competing Interests: Conflict of Interest: No potential conflict of interest relevant to this article was reported.

Background: Oxidative degradation of human recombinant erythropoietin (hrEPO) may occur in manufacturing process or therapeutic applications. This unfavorable alteration may render EPO inefficient or inactive. We investigated the effect of methionine/54 oxidative changes on the amino acid sequences, glycoform distribution and biological activity of hrEPO., Methods: Mass spectrometry was applied to verify the sequence and determine the methionine oxidation level of hrEPO. Isoform distribution was studied by capillary zone electrophoresis method. In vivo normocythemic mice assay was used to assess the biological activity of three different batches (A, B, and C) of the proteins., Results: Nano-LC/ESI/MS/MS data analyses confirmed the amino acid sequences of all samples. The calculated area percent of three isoforms (2-4 of the 8 obtained isoforms) were decreased in samples of C, B, and A with 27.3, 16.7, and 6.8% of oxidation, respectively. Specific activities were estimated as 53671.54, 95826.47, and 112994.93 mg/mL for the samples of A, B, and C, respectively., Conclusion: The observed decrease in hrEPO biological activity, caused by increasing methionine oxidation levels, was rather independent of its amino acid structure and mainly associated with the higher contents of acidic isoforms.

The present study aimed to evaluate in vitro cross-strain inhibitory effects of IgY polyclonal antibody on both growth and urease enzyme of four local strains of H. pylori. Leghorn chickens were immunized with whole cells of four different strains of H. pylori, separately. Rising of specific IgY was detected by ELISA. The IgY purified using polyethylene glycol method and the purity was evaluated by SDS-PAGE and Western blotting. Each strain was treated with its own-specific and also other strain-specific IgYs. The strain-specific IgY could inhibit the growth of specific strains by 49-72% and also other different strains of H. pylori by 29-86%. Our findings revealed that strain-specific IgY could inhibit urease activity of its own by 64-72% and other different strains by 49-79%. These findings confirmed strain-specific and also cross-strain inhibitory effects of the IgY polyclonal antibody on both growth and urease activity of H. pylori., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background and Objective: Some studies suggest that psychological condition and stress can play role in the development of recurrent aphthous stomatitis (RAS). The purpose of this study was to evaluate salivary cortisol changes and psychological conditions in patients with RAS., Materials and Methods: Twenty-seven patients (13 males and 14 females, mean age of 32.8 (±10.2) years) with minor RAS and 27 age- and sex-matched controls without RAS participated in this study. The concentration of cortisol (nanomole/L) was measured in samples of unstimulated saliva from patients and controls two times; once during the presence of active lesions and once again when the lesions had healed by immunologic assay. The Hospital Anxiety and Depression Scale was employed to determine psychological condition. Visual analog scale for pain severity was recorded for patients with active lesions episode. Data were analyzed by the SPSS software (version 18.0) using paired and unpaired t -tests and Pearson correlation coefficient., Results: Salivary cortisol level was lower in patients during active lesions (12.4 ± 5.1) and healing (10.5 ± 3.9) episodes compared to the controls (13.1 ± 3.6) ( P = 0.583, P = 0.015; respectively). There was no significant difference in salivary cortisol between active lesions and healing episodes ( P = 0.943). Anxiety and depression represented no significant differences between active lesions and healing episodes ( P > 0.05). Anxiety and depression levels in patients were significantly higher than in controls ( P < 0.05). Pain severity in active lesions was not significantly correlated to salivary cortisol level, and anxiety or depression scores ( P > 0.05)., Conclusion: The findings showed that occurrence of RAS was associated with anxiety and depression but not with alterations of salivary cortisol level., Competing Interests: There are no conflicts of interest.

Erythropoietin (EPO) is the principal hormone which, has somewhat short half-life involved in the differentiation and regulation of circulating red blood cells. The present study was carried out to evaluate the capability of a polyethylene glycol mimetic technology as a biological alternative to improve pharmaceutical properties of human recombinant EPO. In silico models of EPO fused to 200 amino acids of proline, alanine, and serine (PAS) were initially generated and assessed by molecular dynamic (MD) simulation. The fluctuations of the modeled structure reached a plateau after 6000 ps of MD simulation. The Phi and psi analysis showed >99.2% of residues were located in the allowed regions. An expression vector consisting of EPO cDNA tagged to PAS coding sequences was synthesized and expressed in CHO-K1 Cells. The produced PASylated molecule was purified and characterized by standard analytical methods. The molecular weight of fusion protein was expanded to 70 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis method. Analytical size exclusion chromatography revealed an approximately sevenfold increase in apparent size of produced protein. Although the in vitro potency of the fusion protein was significantly reduced (1.26 ± 0.05 vs. 0.24 ± 0.03 ng/ml) but, the in vivo activity was considerably increased up to 1.58 × 10 5 IU/ml in normocythemic mice assay. Pharmacokinetic animal studies revealed strongly 15.6-fold plasma half-life extension for the PASylated EPO (83.16 ± 13.28 h) in comparison to epoetin α (8.5 ± 2.4 h) and darbepoetin α (25.3 ± 2.2h).

Background: The Copenhagen Psychosocial Questionnaire (COPSOQ) is a widely used tool for evaluation of psychosocial risk factors at work. The aims of this study were to describe the short version of Farsi COPSOQ and to present its psychometric properties. Methods: A total of 427 administrative health care staff participated in this descriptive methodological study. Forward-backward procedure was adopted to translate the questionnaire from English into Farsi. Content validity was assessed by a panel of 10 experts. Construct validity was evaluated by exploratory and confirmatory factor analyses. The internal consistency and test-retest reliability were assessed using Cronbach's α and intraclass correlation coefficient(ICC), respectively. The feasibility was assessed using ceiling and floor effect. Results: The short version of Farsi COPSOQ was configured with 16 dimensions (32 items).Content validity of the questionnaire was established. Factor analysis supported the conceptual multi-dimensionality (four factors), and therefore confirmed the construct validity of the Farsi COPSOQ. The internal consistency (Cronbach's α ranging between 0.75 and 0.89) and test retest reliability (ICC values ranged from 0.75 to 0.89) were both approved and the results showed no ceiling or floor effect. Conclusion: The results support the use of Farsi COPSOQ for evaluation of psychological risks and for research purposes in Iranian population.

Many microfluidic systems-including chemical reaction, sample analysis, separation, chemotaxis, and drug development and injection-require control and precision of solute transport. Although concentration levels are easily specified at injection, pressure-driven transport through channels is known to spread the initial distribution, resulting in reduced concentrations downstream. Here we document an unexpected phenomenon: The channel's cross-sectional aspect ratio alone can control the shape of the concentration profile along the channel length. Thin channels (aspect ratio << 1) deliver solutes arriving with sharp fronts and tapering tails, whereas thick channels (aspect ratio ~ 1) produce the opposite effect. This occurs for rectangular and elliptical pipes, independent of initial distributions. Thus, it is possible to deliver solute with prescribed distributions, ranging from gradual buildup to sudden delivery, based only on the channel dimensions., (Copyright © 2016, American Association for the Advancement of Science.)

Background and Objectives: Numerous procedures in biology and medicine require the counting of cells. Direct enumeration of Colony Forming Units (CFUs) is time-consuming and dreary accurate cell counting on plates with high numbers of CFUs is error prone. In this study we report a new indirect cell counting method that was developed based on the use of Redsafe fluorometric assay. The usefulness of Redsafe, a nucleic acid stain, in liquid medium is based on the binding of the fluorescent dye to DNA., Materials and Methods: Redsafe fluorometric assay was evaluated in comparison with MTT colorimetric assay as a colourimetric assay for enumeration of bacterial cells., Results: Obtained results showed that fluorometric assay threshold for LB grown E. coli is 6×10(4) CFU/ml. Redsafe fluorescent assay can be used as a rapid and inexpensive method for bacterial enumeration and quantification with increased sensitivity., Conclusion: The sensitivity of the Redsafe fluorometric assay for detection and enumeration of bacterial cells was 2-log-unit more than that was observed for the MTT assay.

We study the role geometry plays in the emergence of asymmetries in diffusing passive scalars advected by pressure-driven flows in ducts and pipes of different aspect ratios. We uncover nonintuitive, multi-time-scale behavior gauged by a new statistic, which we term "geometric skewness" S^{G}, which measures instantaneously forming asymmetries at short times due to flow geometry. This signature distinguishes elliptical pipes of any aspect ratio, for which S^{G}=0, from rectangular ducts whose S^{G} is generically nonzero, and, interestingly, shows that a special duct of aspect ratio ≈0.53335 behaves like a circular pipe as its geometric skewness vanishes. Using a combination of exact solutions, novel short-time asymptotics, and Monte Carlo simulations, we establish the relevant time scales for plateaus and extrema in the evolution of the skewness and kurtosis for our class of geometries. For ducts limiting to channel geometries, we present new exact, single-series formulas for the first four moments on slices used to benchmark Monte Carlo simulations.

Tetanus, as a major cause of death in developing countries, is caused by tetanus neurotoxin. Recombinant antibodies against tetanus neurotoxin can be useful in tetanus management. Phage display of antibody fragments from immune human antibody libraries with single chain constructs combining the variable fragments (scFv) has been one of the most prominent technologies in antibody engineering. The aim of this study was the generation of a single chain fragment of variable region (scFv) library and selection of specific antibodies with high affinity against tetanus toxin. Immune human single chain fragment variable (HuscFv) antibody phagemid library was displayed on pIII of filamentous bacteriophage. Selection of scFv clones was performed against tetanus toxin antigens after three rounds of panning. The selected scFv clones were analyzed for inhibition of tetanus toxin binding to ganglioside GT1b. After the third round of panning, over 35 HuscFv phages specific for tetanus toxin were isolated from this library of which 15 clones were found to bind specifically to tetanus toxin. The selected HuscFv phages expressed as a soluble HuscFv peptide and some clones showed positive signals against tetanus toxin. We found that six HuscFv clones inhibit toxin binding to ganglioside GT1b. These selected antibodies can be used in the management of tetanus.

Background: The comparison of Mycobacterium tuberculosis bacterial genotypes with phenotypic, demographic, geospatial and clinical data improves our understanding of how strain lineage influences the development of drug-resistance and the spread of tuberculosis., Methods: To investigate the association of Mycobacterium tuberculosis bacterial genotype with drug-resistance. Drug susceptibility testing together with genotyping using both 15-loci MIRU-typing and spoligotyping, was performed on 2,139 culture positive isolates, each from a different patient in Lima, Peru. Demographic, geospatial and socio-economic data were collected using questionnaires, global positioning equipment and the latest national census., Results: The Latin American Mediterranean (LAM) clade (OR 2.4, p<0.001) was significantly associated with drug-resistance and alone accounted for more than half of all drug resistance in the region. Previously treated patients, prisoners and genetically clustered cases were also significantly associated with drug-resistance (OR's 2.5, 2.4 and 1.8, p<0.001, p<0.05, p<0.001 respectively)., Conclusions: Tuberculosis disease caused by the LAM clade was more likely to be drug resistant independent of important clinical, genetic and socio-economic confounding factors. Explanations for this include; the preferential co-evolution of LAM strains in a Latin American population, a LAM strain bacterial genetic background that favors drug-resistance or the "founder effect" from pre-existing LAM strains disproportionately exposed to drugs.