Evaluation of Microbiological Culture and Antibiogram of the Patient with Corneal Ulcer due to Exposure Keratitis in the Intensive Care Unit in Sina Farshchian Hospital (2019-2020)

Background: Since patients admitted to the intensive care unit have a compromised im-mune system and are more prone to infection than other patients, timely diagnosis and treatment of corneal ulcers among this group of patients can prevent vision loss. Therefore, it is necessary to treat eye infections and corneal ulcers promptly and economize prohibitive costs. Objective(s): Appropriate treatment with the most effective antibiotic before the answer is available to prevent corneal ulcer complications and blindness. Method(s): This study was conducted from November 2019 to November 2020 and after approval by the ethics committee of Hamedan University of Medical Sciences with the code of ethics: IR.UMSHA.REC.1398.716. First, the corneal secretions of 121 patients admitted to the intensive care unit of Sina Hospital are prepared by an ophthalmologist (after anesthetizing the cornea with tetra-caine drops and sterile swabs) and culture in four growth mediums (blood agar, chocolate agar, thio-glycolate, and EMB). Microbial cultures are examined after 48 hours and a fungal culture is examined one week later. Disc diffusions are placed in positive microbial cultures. Antibiotic susceptibility or resistance of the antibiogram was recorded. Other demographic data, including patients' age and sex, are extracted from ICU files. Also, test results and patient identifications are recorded in a checklist designed for this purpose. Result(s): Of all the antibiotics used against common bacteria, vancomycin (84%), colistin (80.43%), cefazolin (80%), and levofloxacin (60%) had the highest sensitivity and gentamicin (93.75%), ceftazidime (86.42%) Erythromycin (85%) had the highest resistance against isolated bacteria. Conclusion(s): The data obtained from this study showed that the most common microorganisms in the age group under the age of 30 years were Acinetobacter Baumannii, in the group of 30-60 years old was Klebsiella pneumonia, and age group over 61 years old was Staphylococcus aureus, and the most sensitive antibiotics in the age group under 30 years were vancomycin and levofloxacin and the age group30-60 were colistin and vancomycin and in the age group over 61 years were vancomycin and cefazolin.Copyright © 2023 Bentham Science Publishers.

Analysis of approaches to sterility testing of COVID-19 prevention vaccines.

Preventive vaccination against SARS-CoV-2 infection is currently receiving close attention in the Russian Federation. Improving public confidence in immunisation with new vaccines largely depends on a guarantee of the absence of side effects caused by contamination. A high risk of contamination is inherent to biological products, including coronavirus prevention vaccines, due to their properties and the nature of raw materials used. This risk adds to the need for using effective contaminant detection approaches. The aim of the study was to evaluate the possibility to improve sterility testing of preventive vaccines against SARS-CoV-2 infection. This article presents an analysis of the procedures proposed by pharmaceutical developers for sterility testing of ten Russian vaccines approved in the country for COVID-19 prevention. The authors considered specific characteristics of these vaccines, including their physical and chemical properties, the presence of antimicrobial components, and other critical factors affecting the correctness of the experimental setup. The results suggest that it is possible to improve sterility testing. According to the authors, the main directions for its improvement are the proposal to develop an alternative procedure based on compendial method 2 (OFS.1.2.4.0003.15, Ph. Rus. XIV), as well as the use of a universal culture medium. If used for refining the established procedures and developing new ones, the authors' recommendations will improve the reliability and applicability of sterility testing during both manufacturing and pre-approval regulatory assessment of updated coronavirus vaccines for subsequent release to the market. The proposed approaches can be applied to testing other medicinal products for sterility.Copyright © 2023 National Electronic-Information Consortium (NEICON). All rights reserved.

EWRI Friends Visit London Water & Steam Museum and Union Chain Bridge-IHCEL

We strongly recommend EWRI members visit the London Water & Steam Museum. This presentation includes many slides of this outstanding museum near the Kew tube station. London Museum of Water & Steam features artefacts and interactive exhibits on water. On October 26, 2018, David Gilbert and Jerry Rogers toured the London Museum of Water & Steam, located at Green Dragon Lane, Brentford, London, TW8 0EN near Kew station. Kew Bridge Pumping Station was originally opened in 1838 by the Grand Junction Waterworks Company. In 1999, the United Kingdom government's Department for Culture, Media, and Sport described Kew Bridge Pumping Station as "the most important historic site of the water supply industry in Britain." The heart of the museum showcases a majestic collection of steam pumping engines, including engines from Corynwall, as well as rotative engines. There are many excellent London water supply and treatment exhibits also. Due to COVID-19, the planned International Historic Civil Engineering Landmark plaque ceremony of July 26, 2020, for the 200-year-old Union Chain Suspension Bridge at Berwick-upon-Tweed was cancelled. Note the book: Samuel Brown and Union Chain Bridge: Gordon Miller, Friends of the Union Bridge, 306 pp, 135 photographs, 15.5 GBP. A tour of the Paxton Estate (Paxton Trust), a historic house at Paxton, Berwickshire, was planned to be a part of the plaque ceremony. There is discussion of having a modified plaque ceremony in the spring of 2023 (specific date to be determined) possibly before the May 2023 EWRI Congress in Henderson, Nevada. © World Environmental and Water Resources Congress 2023.All rights reserved

Gene Editing/Gene Therapies: BIOREACTOR RECOMBINANT PRODUCTION OF SARS-COV-2 VIRUS ANTIGENS IN CHO CELL CLONES BY USING NEW UCOE LENTIVIRAL VECTOR SYSTEM

Background & Aim: The new UCOE models we have recently developed, tested on many cell groups (including mouse ES and human iPS cells) and human mAb recombinant production studies as well, shows a powerful resistance to DNA methylation- mediated silencing and provides a higher and stable transfection profile. By the urgent need of vaccine development for COVID-19 during the pandemic, in this study we aimed to produce a potential recombinant vaccine by using the new generation UCOEs models of our own design. Methods, Results & Conclusion(s): Existing new-generation UCOE models and standard plasmid vectors to be used as control group were provided. Then, the sequences related to the PCR method were amplified for sufficient stock generation and cloning experiments. Verification in the plasmid vector was carried out in gel electrophoresis. Transfection of 293T cells was performed with clone plasmids carrying antigen genes and plasmids carrying genetic information of lentivirus units for the production of lentiviral vectors. Afterwards, 293T cells produced lentiviral vectors carrying antigen genes. Harvesting of these vectors was carried out during 48th and 72nd hours. Afterwards, CHO cells were transduced with appropriate quantity of lentiviral vectors. Isolation and purification of targeted proteins from the relevant medium were performed by HPLC and Q-TOF methods. A part of the spike and nucleocapsid gene sequences of COVID-19 were firstly cloned into our UCOE models. These UCOEs plasmids were then transferred into 293T cells along with plasmids carrying the genes that will form the lentivirus vectors (LVs). After harvesting and calculation of LV vector titers, the cloned vectors were then transfected into the CHO cells which the targeted recombinant production of the antigen proteins will be carried out. Antigenic structures were then isolated from the culture medium of CHO cells in following days for confirmation. Using HPLC and qTOF mass spectrometer methods, these structures in the medium were confirmed to be the units of spike and nucleocapsid proteins of the COVID-19 virus. In order to produce large amount of the recombinant antigens, the culture was then carried out with bioreactors in liters. At the final stage, these recombinantly produced antigen proteins were tested on rats to measure their immunogenic responses, and the study recently been completed successfully as a potential recombinant vaccine against COVID-19.Copyright © 2023 International Society for Cell & Gene Therapy

Analysis of approaches to sterility testing of COVID-19 prevention vaccines

Preventive vaccination against SARS-CoV-2 infection is currently receiving close attention in the Russian Federation. Improving public confidence in immunisation with new vaccines largely depends on a guarantee of the absence of side effects caused by contamination. A high risk of contamination is inherent to biological products, including coronavirus prevention vaccines, due to their properties and the nature of raw materials used. This risk adds to the need for using effective contaminant detection approaches. The aim of the study was to evaluate the possibility to improve sterility testing of preventive vaccines against SARS-CoV-2 infection. This article presents an analysis of the procedures proposed by pharmaceutical developers for sterility testing of ten Russian vaccines approved in the country for COVID-19 prevention. The authors considered specific characteristics of these vaccines, including their physical and chemical properties, the presence of antimicrobial components, and other critical factors affecting the correctness of the experimental setup. The results suggest that it is possible to improve sterility testing. According to the authors, the main directions for its improvement are the proposal to develop an alternative procedure based on compendial method 2 (OFS.1.2.4.0003.15, Ph. Rus. XIV), as well as the use of a universal culture medium. If used for refining the established procedures and developing new ones, the authors' recommendations will improve the reliability and applicability of sterility testing during both manufacturing and pre-approval regulatory assessment of updated coronavirus vaccines for subsequent release to the market. The proposed approaches can be applied to testing other medicinal products for sterility.Copyright © 2023 National Electronic-Information Consortium (NEICON). All rights reserved.

Sars-Cov-2 Vaccine from Infected Plant Attenuated Virus Using Acer Cincinatum (Vine Maple)

Purpose of study Proposal for an oral (or if required, parenteral) COVID-19 vaccination based upon this described technology. Investigational theory under study for the past 9 months of COVID-19 growing season. Coronavirus can attack and infect plant species. It was found that SARS-CoV-2 can infect various plant species. Others have found plants, for example tobacco as a good growth medium for Coronavirus and SARS-CoV-2. This current study has found various plants species infected with SARS-CoV-2 by rPCR. As the plants were located beside a well used hiking trail for humans, and were infected along the trail including various species with SARS-CoV-2, hypothesized that human airborne contact had caused infection in the bordering plants. Humans were observed to be coughing while walking on the trail, and were not wearing masks. The plant leaves developed small circular colonies of the virus, which became self-limited at several millimeters in diameter. All of the plants were clear of these lesions before the COVID-19 Pandemic. The plants 'immune' system produced antiviral agents, including lectins which limited the growth of the colonies and prevent death of the leaf and whole plant. The fungal cultures of the 'spots' were negative. The rPCR of all spots tested in the present series was positive for SARS-CoV-2. Hypothesis, that self-augmentation of the virus occurred by the natural culturing in plant leaves that produce antiviral agents as part of their 'immune system.' Hypothesis, a symbiotic type relationship developed between the plant using its chemical immune system, and the virus allowed to replicate in an augmented fashion to allow both the virus and the host to survive and grow. As the top candidates for the oral vaccine are nontoxic, hypothesis involves the maceration of the infected leaves, mixing with a nontoxic adjuvant and flavoring to promote assimilation and palatability, with the proposed route of entry being mastication, thus exposing the oral-nasal mucosa to the vaccine, with the probable best of immunity to usual exposure to the SARS-CoV-2 virus, that is the oral-nasal mucosal and upper airway route. As many types of animals are now infected with SARS-CoV-2, it is further hypothesized that this oral vaccine could also be mass produced to add to various animals by feedstock and oral route. Methods used Hypotheses formed through observations. Testing of observations by pPCR, viral cell culture, fungal culture, light and electron microscopy. Summary of results pPCR SARS-CoV-2 positive, cell culture 'lysis experiment' positive, EM and light microscopy positive, fungal culture negative. Conclusions TABLE OF HYPOTHESES AND STUDY RESULTS (HYPOTHETICAL, OBSERVED, PROVEN) 1. The first hypothesis that the virus is attenuated by the plant, using its innate chemical immune system. Similarly, Pasteur used chemical such as phenol to attenuate viruses for wome of the first successful vaccines. Observed. 2. Hypothesis, the plants 'immune' system produced antiviral agents, including lectins, flavonoids, and others, which limited the growth of the colonies and prevent death of the leaf and whole plant. Proven. 3. Hypothesis is that the nontoxic plants, such as Vine Maple sp.(Acer cincinatum), could be used to produce and oral plant attenuated vaccine. Hypothesis. 4. Hypothesis involves the maceration of the infected leaves, mixing with a nontoxic adjuvant and flavoring to promote assimilation and palatability, with the proposed route of entry being mastication, thus exposing the oral-nasal mucosa to the vaccine, with the probable best of immunity to usual exposure to the SARS-CoV-2 virus, that is the oral-nasal mucosa, upper airway. (Figure Presented).

Centella asiatica extract inhibits Mycobacterium tuberculosis growth in rat tuberculosis models

Background and Aims: The covid-19 pandemic has reversed years of progress in the fight to end tuberculosis. So, the discovery of new drugs as antituberculosis is very much needed. Our previous studies have shown that the extract of Centella asiatica is able to inhibit the growth of Mycobacterium tuberculosis in vitro and requires further research. The aims of this study is to prove the effect of Centella asiatica inhibit Mycobacterium tuberculosis in rat model tuberculosis. Method(s): The protocol in this study was approved by the veterinary ethics committee of Airlangga University. The rat tuberculosis model was induced by intrathecal injection of a suspension of Mycobacterium tuberculosis strain H37 Rv. Twenty-eight tuberculosis rat were randomly divided into four groups. Groups 1,2, and 3 were treated with ethanol extract of Centella asiatica at 375 mg/kgBW, 750 mg/kgBW and 1500 mg/kgBW, and the fourth group was the control group. Centella asiatica extract is administered orally via an intragastric feeding tube for two weeks, once daily At the end of the experimental period, rats were sacrificed by cervical decapitation. The left lung tissue was taken aseptically and cultured on Middlebrook 7H10. Result(s): The results showed that there was no bacterial growth on the culture media in the group that received Centella asiatica extract at a dose of 750 and 1500 mg/kg BW. Conclusion(s): The conclusion in this study, that Centella asiatica extract inhibit the growth of Mycobacterium tuberculosis at doses of 750 and 1500 mg/kg BW. We thank the Directorate of Research and Community Services, the Directorate General of Higher Education, and Ministry of Education and Culture in Indonesia for the financial supportCopyright © 2023 The American Society for Biochemistry and Molecular Biology, Inc.

Surfactant Protein B Reduces Respiratory Viral Infectivity in Human Lungs

Purpose of Study: COVID pneumonia caused by SARS-CoV-2 can result in a depletion of surfactant & lung injury, which resembles neonatal respiratory distress syndrome. Exogenous surfactant has shown promise as a therapeutic option in intubated hospitalized patients. Our preliminary data in human lung organoids (LOs) with a deficiency of surfactant protein B (SP-B) showed an increased viral load compared to normal LOs. Single cell RNA sequencing (scRNAseq) revealed that SP-B-deficient cells showed increased viral entry genes (ACE2 receptor) & dysregulated inflammatory markers emanating from the lung cells themselves. Our objective was to determine: (1) cell-specific transcriptional differences between normal & SP-B deficient human lung cells after infection with SARS-CoV-2 and (2) a therapeutic role of SP-B protein & surfactant in COVID-19 pneumonia. Methods Used: We used normal and SP-B mutant (homozygous, frameshift, loss of function mutation p.Pro133GlnfsTer95, previously known as 121ins2) human induced pluripotent stem cells (hiPSC) and differentiated them into 3D proximal lung organoids. The organoids were infected with the delta variant of SARS-CoV-2 for 24 hours at an MOI of 1. Infected and uninfected organoids were fixed in trizol in triplicate and underwent processing for bulk RNA sequencing. We tested for differentially expressed genes using the program DEseq. We also plated normal iPSC derived lung organoids as a monolayer and pre-treated them with 1mg/ml of Poractant alfa or 5 uM of recombinant SP-B protein. The delta strain of SARS-CoV-2 was added to the 96 wells at an MOI of 0.1 for one hour with shaking, then an overlay with DMEM/CMC/FBS was added and left on for 23 hours. The plate was fixed and stained for nucleocapsid (NC) protein. Summary of Results: Bioinformatic analysis of the bulk RNA sequencing data showed an increase in the multiple cytokines and chemokines in the SP-B mutant LOs compared to control. We also saw differential gene expression patterns in the SP-B mutant LOs including a reduction in SFTPC, FOXA2, and NKX2-1 and an increase in IL1A, VEGFA, PPARG and SMAD3. In the exogenous surfactant experiments, there was a decrease in total expression of viral NC in the Poractant alfa & rSP-B-treated cells compared to SARS-CoV-2 infection alone (p<0.001). Conclusion(s): Surfactant modulates the viral load of SARS-CoV-2 infection in the human lung. Deficiency in SP-B results in the dysregulation of the lung epithelial inflammatory signaling pathways resulting in worsening infections.

Comparison of Perindopril vs. Losartan With Regard to Pro-Coagulation Factors in Adipocytes Exposed to Sars-Cov-2 Spike Protein

Background: Obesity is a strong risk factor for more severe Covid-19 infection as adipocytes play an important role in intermediating the spreading, replication, and release of SARS-COV-2. An increase in pro-coagulation factors (tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1)) was observed in Covid-19 patients with moderate to severe symptoms and is reported to be associated with Angiotensin-converting enzyme 2 (ACE2) overexpression. Cardiovascular medications affecting ACE2, such as Perindopril and Losartan, are hypothesized to have an effect on Covid-19 infection-related coagulopathy. This study aims to identify and compare the effect of perindopril and losartan on TF and PAI-1 levels in adipocytes exposed to SARS-COV-2 spike protein. Method(s): Adipocytes were isolated enzymatically from adipose tissue obtained from an obese male donor. Adipocytes were then exposed to SARS-COV-2 S1 spike protein for 24 hours. After exposure, perindopril and losartan were added to the culture medium. ACE2, TF, and PAI-1expression were measured 2 hours later using ELISA. Result(s): SARS-CoV-2 spike protein exposure increased ACE2, TF, and PAI-1 expression. Perindopril addition discernible reduced the tissue factor (TF) expression (4.843 +/- 0.396) compared to a positive control (6.857 +/- 0.228) (p=0.005) but not losartan (5.624 +/- 0.606) (p=0.111). Perindopril was also able to lower PAI-1 expressions (3.484 +/- 0.252) compared to a positive control (4.865 +/- 0.115) (p=0.001), but the losartan did so more effectively (2.633 +/- 0.269) (p=0.000). Conclusion(s): Losartan and perindopril both have the ability to lower pro-coagulation factors, proving the value of ACEIs/ARBs in preventing thrombotic complications in Covid-19 patients.Copyright © 2023

Protease Activity in the Excretory-Secretory Products of Nymphal Stage of Linguatula Serrata: First Detection of a Metalloprotease

The aim of this study was to evaluate the presence of proteases and determine the main protease present in the excretory-secretory products (ESPs) from nymphal stage of Linguatula serrata. Infected mesenteric lymph nodes of goats were collected from Tabriz slaughterhouse, northwestern Iran. Recovered Linguatula serrata nymphs were immersed in culture medium (MEM), then ESPs were collected and protease activity in presence of specific inhibitors was assayed. Protease enzyme was fur-ther characterised by SDS-PAGE. The results of this study showed that the main protease in the ESPs from the nymphal stage of L. serrata was a metalloprotease that was resistant to heat. In conclusion, these data show that a major protease secreted by the larval stage of L. serrata exhibited properties that may play a role in the pathogenesis of L. serrata nymphs.Copyright © 2023, Trakia University. All rights reserved.

Viral enteritis in cattle: To well known viruses and beyond

Livestock products supply about 13 percent of energy and 28 percent of protein in diets consumed worldwide. Diarrhea is a leading cause of sickness and death of beef and dairy calves in their first month of life and also affecting adult cattle, resulting in large economic losses and a negative impact on animal welfare. Despite the usual multifactorial origin, viruses are generally involved, being among the most important causes of diarrhea. There are several viruses that have been confirmed as etiological agents (i.e., rotavirus and coronavirus), and some viruses that are not yet confirmed as etiological agents. This review summarizes the viruses that have been detected in the enteric tract of cattle and tries to deepen and gather knowledge about them.Copyright © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Effects of physicochemical changes (temperature, pH, and culture media) on strong biofilm formation of Acinetobacter baumannii isolated from patients with respiratory infection in Iraq

Introduction and Aim: The ability of Acinetobacter baumannii to form biofilms on biotic and abiotic surfaces is regulated by several pathogens' virulence factors, and this is thought to be at the root of the bacteria's resistance to antibiotics. We hope to learn how temperature, pH, and iron concentrations influence the development of biofilms in A. baumannii isolated from COVID-19 and non-COVID-19 individuals, and which genes are relevant for biofilm formation and antibiotic resistance. Material(s) and Method(s): Eight strong adherent isolates of A. baumannii from respiratory tract infection Iraqi patients (4 from COVID-19 and the other from non-COVID-19 just respiratory patients) had been used in this study which conducted from 10/1/2021 to 10/2/2022. The antibiotic sensitivity of all isolates was determined using the VITEK-2 system. The biofilm associated genes OXA-51, bap, Chaperone Usher (CsuE) and Integron-1, was detected using PCR. Isolates of A. baumainni were put through a battery of tests to determine whether they possessed the capacity to produce robust biofilms under a wide range of both physical (temperature, pH) and chemical circumstances. Result(s): A. baumannii showed that all isolates were multidrug resistant and positive for the biofilm genes studied. Effect on temperature on biofilm formation showed at 44C biofilm formation was significantly lower than that at 37C (mean differences of 0.178000 (t= 8.355, df:3, P=0.004) and 0.204000 (t=26.521, df:3, P=0.000) respectively). The adhesion factor value in the COVID-19 positive and negative groups decreased significantly because of the pH change. Iron concentration of 60 microM significantly lowered biofilm formation among COVID-19 group and non-COVID-19 group. Conclusion(s): A. baumanni are multidrug resistance isolates with a capacity to form biofilms. The ability to form biofilms by A. baumannii is strongly influenced by physical and chemical factors.Copyright © 2023, Indian Association of Biomedical Scientists. All rights reserved.

The SpACE-CCM: A facile and versatile cell culture medium-based biosensor for detection of SARS-CoV-2 spike-ACE2 interaction.

The COVID-19 pandemic is an ongoing global public health threat. COVID-19 is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, and binding of the SARS-CoV-2 spike to its receptor, angiotensin-converting enzyme 2 (ACE2), on host cells is critical for viral infection. Here, we developed a luminescent biosensor that readily detects interactions of the spike receptor-binding domain (RBD) and ACE2 in cell culture medium ('SpACE-CCM'), which was based on bimolecular complementation of the split nanoluciferase-fused spike RBD and ectodomain of ACE2 and further engineered to be efficiently secreted from cells by adding a heterologous secretory signal peptide (SSP). Screening of various SSPs identified 'interferon-α+alanine-aspartate' as the SSP that induced the highest activity. The SpACE-CCM biosensor was validated by observing a marked reduction of the activity caused by interaction-defective mutations or in the presence of neutralizing antibodies, recombinant decoy proteins, or peptides. Importantly, the SpACE-CCM biosensor responded well in assay-validating conditions compared with conventional cell lysate-based NanoLuc Binary Technology, indicating its advantage. We further demonstrated the biosensor's versatility by quantitatively detecting neutralizing activity in blood samples from COVID-19 patients and vaccinated individuals, discovering a small molecule interfering with the spike RBD-ACE2 interaction through high-throughput screening, and assessing the cross-reactivity of neutralizing antibodies against SARS-CoV-2 variants. Because the SpACE-CCM is a facile and rapid one-step reaction biosensor that aptly recapitulates the native spike-ACE2 interaction, it would be advantageous in many experimental and clinical applications associated with this interaction.

Negative Urine Legionella Antigen Does Not Rule out Legionella Pneumonia: A Case Report

INTRODUCTION: Legionella is an important cause of community acquired pneumonia. Here we describe a case with strong clinical suspicion of Legionella pneumonia despite negative urine antigen but confirmed by polymerase chain reaction (PCR) test on a lower respiratory sample. DESCRIPTION: A 52-year-old male, nonsmoker with unremarkable past medical history presented with a 4-day history of fever, nonproductive cough, and malaise, after returning from a trip to Italy. He underwent Computed Tomography Angiography (CTA) chest which revealed bilateral lung infiltrates at Urgent Care. Both Rapid Ag and PCR test for SARS CoV-2 were negative. He was vaccinated against SARS-CoV2. On hospital admission his oxygen saturation was 85% on room air. Lab work revealed white blood cell (WBC) 22.5, Hemoglobin 12.8, Platelets 397 with 96% neutrophils. His Sodium was 135 mmol/L (135- 146), CRP 433.2 mg/L (normal < 5). Respiratory PCR was negative for Influenza A, B, RSV. Urine Pneumococcal and Legionella Ag were negative. He was started on Ceftriaxone and Azithromycin. He developed rapidly progressing respiratory failure leading to intubation, prone positioning, inhaled Prostacyclin due to significant hypoxia (P/F ratio 57). His antibiotics were changed to high dose Levofloxacin (750 mg IV Q Day) because of strong suspicion of Legionella. He underwent bronchoscopy with BAL and the PCR came back positive for Legionella. The patient was extubated in 48 hours and discharged home after a 10-day course of Levaquin. The BAL sample was sent to Centers for Disease Control and Prevention (CDC) which identified Legionella species as serotype 1. DISCUSSION: Legionella are gram negative facultative intracellular bacteria with soil and water as reservoirs. Legionella grows poorly on routine culture media. Urine antigen (Ag) testing has a sensitivity of 75% and detects Legionella pneumophilia serotype 1, the dominant cause (80% of cases) but does not detect the other 30 Legionella species that have been isolated from humans. A lower respiratory PCR detects other serotypes and perhaps is more sensitive than urine Ag in detecting serotype 1. If clinical suspicion of Legionella is high PCR must be performed on a lower respiratory sample and one must not solely rely on a negative Urinary Ag test.

Interleukins-a non-invasive biomarker for invasive fungal sinusitis

Objectives: Invasive fungal infection is estimated to cause around 1.5 million deaths each year. But the true burden is estimated to be even more due to the lack of reliable diagnostic methods.Invasive fungal sinusitis (IFS) is the emerging infection leading to high morbidity and mortality (80%). Both clinical suspicion and reliable diagnostic tests available to diagnose an IFS are less. As a result, most patients are diagnosed only in the later stages. Nasal swabs for IFS are less useful and the most appropriate sample for the diagnosis will be the deep tissues involved, which are obtained after an invasive procedure.The gold standard test for diagnosis of IFS is the isolation of fungus in culture media but its sensitivity is 50%. Hence a non-invasive procedure to help in the diagnosis of invasive fungal sinusitis is needed. The role of Interleukins (IL) in fungal infections, especially IL-10 and IL-17 has been documented in various research in the past. Hence this research was carried out to study the role of IL-10 and IL-17 in invasive and non-invasive fungal sinusitis (NIFS). Method(s): The study wascarried out in the Department of Microbiology, SRIHER, Chennai. Atotal of 60 samples collected from patients suspected to have fungal sinusitis and sent to the laboratory for fungal culture were considered for the study. All the samples which grew fungus were categorized as IFS and NIFS. ELISA was performed with the serum samples of patients for IL-10 and IL-17 based on manufacturer's instruction (Human IL-10 ELISA Kit, Therm ofisher and Human IL-17 ELISA Kit, Therm ofisher), and reading is taken in a spectrophotometer (Thermo Fisher Scientific). Result(s): Among the 60 serum samples tested, 30 were categorized as IFS and the rest as NIFS.A total of 90% (n = 27/30) of IFS patients expressed interleukins in serum samples whereas none of the NIFS expressed both the interleukins tested.Among IFS, IL-10 was seen in 63.3% (n = 19/30) patients, IL-17 in 46.7% (n = 14/30) patients and 20% (n = 6/30) patients expressing both IL-10 and IL-17. In IFS the mean value of IL-10 and IL-17 were 6.657 and 4.259 respectively. Among the 30 IFS, 13 were positive for COVID-19. IL-17 was expressed in 84.6% (n = 11/13) of COVID-19 positive IFS patients. But only 23.1% (n = 3/13) of COVID-19 positive IFS patients expressed IL-10. A total of 15.4% (n = 2/13) of the COVID-19 positive patients did not express any interleukins. Surprisingly the expression of IL-10 among COVID-19 negative IFS was 94.1% (n = 16/17). The specificity of both IL-10 and IL-17 was 100% in the case of IFS. Conclusion(s): Thus, interleukins look to be a promising biomarker for IFS. Further studies will help in establishing inter-leukins as a potential non-invasive biomarker for IFS. IL-17 can be used as a biomarker for COVID-19 patients suspected to have IFS.Also looking for more than one cytokine preferably a combination of IL-10 and IL-17 should be done in patients with NIFS, which will help in the early prediction of patients progressing into IFS and can be managed accordingly.

Perspectivas Inovadoras Na Abordagem Na Doenca Do Olho Seco (Dos) E O Interfaceamento Com Bancos De Sangue De Cordao Umbilical Humano

Os bancos de sangue de cordao umbilical e placentario (BSCUP), os laboratorios de processamento de medula ossea/ sangue periferico para transplante e os centros de tecnologia celular, passaram a receber a denominacao comum de Centros de Processamento Celular - CPC. O pais tem BSCUP 14 unidades publicas e 19 de natureza privada, totalizando 33 BSCUP. Uma analise retrospectiva dos relatorios da ANVISA identifica que em 2003 foram coletadas 26 unidades, com desqualificacao de 15,38% delas, o crescimento da coleta foi exponencial e dez anos depois, em 2013, foram coletadas 13.995 unidades (5,82% de desqualificacao). Em 2020, houve uma diminuicao expressiva de coletas, reflexo da pandemia de COVID-19: 4.918 unidades (desqualificacao de 12,69%). Este tipo de produtividade compromete a viabilidade financeira destes servicos, e encontrar formas de otimizar bolsas desqualificadas por volume ou quantidade de celulas para demais finalidades e uma vertente de gestao que deve ser estabelecida. O objetivo deste projeto foi a coleta de segmentos de cordao umbilical das unidades ja previamente validadas pelos BSCUP para extracao de celulas tronco, caracterizacao imunofenotipica e producao de meio condicionado isento de soro fetal bovino. A obtencao do meio condicionado (MC) da cultura de celulas tronco. Tem crescido cada vez mais o interesse pelo uso dos fatores de crescimento, citocinas e moleculas sinalizadoras livres no MC alem das vesiculas extracelulares, que se tornaram relevantes, tanto para diagnostico como para terapeutica, inclusive para aplicacoes oftalmologicas. Neste campo, identificamos a DOS que impacta profundamente a qualidade de vida das pessoas. Ha 15 anos o Laboratorio de Biologia Celular tem desenvolvido o soro autologo, para atendimento dos pacientes refratarios aos tratamentos convencionais e farmacologicos disponiveis, em especial aqueles pacientes submetidos ao Transplante de Medula Ossea e que desenvolveram DOS2ario a doenca de enxerto versus hospedeiro. No entanto, existem pacientes com impossibilidade de acesso venoso ou com sorologias reagentes para doencas infecciosas que sao impedidos de utilizar o produto. Diante disto, optou-se por produzir o MC de celulas tronco de cordao umbilical de parturientes jovens e sem comorbidades para obtencao do secretoma das celulas para avaliacao terapeutica na DOS. Um segmento do cordao umbilical foi retirado e processado seguido de plaqueamento e expansao para posterior identificacao de adesao ao plastico, caracterizacao imunofenotipica por citometria de fluxo utilizando marcadores como CD11b, CD13, CD14, CD34, CD31, CD36, CD45,CD73, CD90, CD 105, CD106 e HLA-DR. Todas as amostras tiveram adesao ao plastico com aspecto fibroblastoides e perfil imunofenpotipico corrobora com o determinado pela SITC. Para a obtencao de MC foram semeadas CTMcup ate 70% de confluencia e foram submetidas ao wash out, recebendo meio de cultura DMEM-F12 aditivado por 48 horas. Apos isto, foi coletado 60mL do secretoma das celulas para o experimento especificos in vitro. Copyright © 2022

Viral Infectivity Inhibition and Viral Biological Elements Destruction using Safe and Low Power Electrons Generated by Life Restoration Device (LRD): An In Vitro Study

Background: Recently, different side effects have been observed after using antiviral drugs before activation of the immune system. Therefore, it is very important to use effective and non-invasive therapy with fewer side effects for infected virus treatment. Method(s): In this study, we designated a new device termed a Life Restoration Device (LRD). The main function of LRD is to generate electric frequencies with lower and safer potential. These frequencies can effectively destroy the biological elements in the viruses, such as nucleic acid materials and viral cell membranes, but not the cellular plasma membrane of the infected eukaryotic cells. Result(s): A designated glass tube was prepared for this purpose. The infected cell culture was located in the cell culture media, and propagated viruses were poured into the glass tube. Additionally, two nickel-coated copper rods were inserted into both ends of the tube inside the cell culture media. Afterward, the two nickel-coated copper rods were connected to the LRD. Using LRD, lower potential electric frequencies were generated and applied for 30 min and 60 min time points. The treatment of the cell culture containing MERS-CoV and SARS-CoV-2 with LRD for 30 min significantly reduced the viral infectivity by 83% and 22%, respectively. After 60 min of treatment with LRD, the infectivity of MERS-CoV and SARS-CoV-2 viruses was reduced by 21% and 1%, respectively. Furthermore, HIV and HBV-infected blood showed a 95.5% and 100% viral inhibition rate after 2 h exposure to LRD. Additionally, based on the results of the electron microscopy of treated H5N1 virus and western blot analysis data of different types of viruses, the nucleic acid components of the treated viruses were reduced compared to the non-treated viruses. The low-power electric frequencies produced by LRD can reduce the fluidity and osmosis of the viral envelope but not the plasma membrane of the infected cells. Conclusion(s): Treatment of different types of pathogenic viruses with electric stimulation produced by LRD is a new alternative to safe therapy but needs further investigations. The results of this study are important to develop an effective, safe, and alternative viral therapy. Copyright © 2022 Bentham Science Publishers.

Plasma-Like Culture Medium for the Study of Viruses.

Viral infections attract more and more attention, especially after the emergence of novel zoonotic coronaviruses and the monkeypox virus over the last 2 decades. Research on viruses is based to a great extent on mammalian cell lines that are permissive to the respective viruses. These cell lines are usually cultivated according to the protocols established in the 1950s to 1970s, although it is clear that classical media have a significant imprint on cell growth, phenotype, and especially metabolism. So, recently in the field of biochemistry and metabolomics novel culture media have been developed that resemble human blood plasma. As perturbations in metabolic and redox pathways during infection are considered significant factors of viral pathogenesis, these novel medium formulations should be adapted by the virology field. So far, there are only scarce data available on viral propagation efficiencies in cells cultivated in plasma-like media. But several groups have presented convincing data on the use of such media for cultivation of uninfected cells. The aim of the present review is to summarize the current state of research in the field of plasma-resembling culture media and to point out the influence of media on various cellular processes in uninfected cells that may play important roles in viral replication and pathogenesis in order to sensitize virology research to the use of such media.

Virus Enrichment in Cerebrospinal Fluid Using VIRRION Technology

Aseptic meningitis and encephalitis contribute to acute and chronic neurological disorders throughout the world and have a greater than 5% fatality rate in the United States. Unfortunately, in the vast majority of cases, no causative organism is isolated using conventional molecular and immunologic assays. This is in part due to assays that require prior knowledge of specific virus(es), need for large sample volumes, and significant time to obtain results which precludes usage in the clinical setting. To address these limitations to effectively detect virus in CSF samples, we aimed to use a novel technology called VIRRION (virus capture with rapid Raman spectroscopy detection and identification). VIRRION is a combined microfluidic device and carbon nanotube array for enrichment and capture of virus particles in body fluids that uses Raman spectroscopy for rapid detection of virus in which machine learning algorithms are applied to spectra to develop virus 'signatures'. While VIRRION has been successfully used for respiratory isolates, including SARS-COV-2, here we propose to extend this system for the analysis of cerebrospinal fluid from patients with neurologic disease in which viruses are suspected. In preliminary results with samples containing viruses in culture media, we have demonstrated Raman spectral peaks specific to each cultured virus where machine learning analysis showed a sensitivity of 99%. VIRRION offers the capability to capture virus particles while allowing the background fluids to flow through, thus controlling for variation in the background fluid signatures in clinical samples. These early results show that VIRRION is a promising technology for the rapid detection of virus in the CSF.

Figurate eczema of the forearm in a hairdresser

Hairdressers are among high risk professions for occupational skin diseases development, and major attention is usually dedicated to contact allergy. However, infections are behind the corner, and should not be undervalued. We present an emblematic case of tinea incognita, for the iconography, typical history of misdiagnosis, mimicking contact eczema and incongruous treatment with corticosteroids. Actual COVID-19 pandemic restrictive measures caused an additional diagnostic delay, blocking the access to outpatient dermatology service. However, the diagnosis is very simple, when direct microscopic examination is available or grace to selective culture medium disposal. The latter do not require incubation, the dermatophytes growing at room temperature in few days, inducing the change in the medium color. Further identification of the dermatophyte species requires a more experienced mycology laboratory, but it is not essential for the treatment choice. Copyright © by Journal of Applied Cosmetology.