ABSTRACT
Previous studies towards reduced oxygen availability have mostly focused on changes in total mRNA expression, neglecting underlying transcriptional and post-transcriptional events. Therefore, we generated a comprehensive overview of hypoxia-induced changes in total mRNA expression, global de novo transcription, and mRNA stability in monocytic THP-1 cells. Since hypoxic episodes often persist for prolonged periods, we further compared the adaptation to acute and chronic hypoxia. While total mRNA changes correlated well with enhanced transcription during short-term hypoxia, mRNA destabilization gained importance under chronic conditions. Reduced mRNA stability not only added to a compensatory attenuation of immune responses, but also, most notably, to the reduction in nuclear-encoded mRNAs associated with various mitochondrial functions. These changes may prevent the futile production of new mitochondria under conditions where mitochondria cannot exert their full metabolic function and are indeed actively removed by mitophagy. The post-transcriptional mode of regulation might further allow for the rapid recovery of mitochondrial capacities upon reoxygenation. Our results provide a comprehensive resource of functional mRNA expression dynamics and underlying transcriptional and post-transcriptional regulatory principles during the adaptation to hypoxia. Furthermore, we uncover that RNA stability regulation controls mitochondrial functions in the context of hypoxia.
Subject(s)
Gene Expression Regulation , Hypoxia , Acclimatization , Humans , Hypoxia/genetics , Hypoxia/metabolism , RNA Stability/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolismABSTRACT
BACKGROUND: Possible cases of SARS-CoV-2 infection were diagnosed in primary care in Madrid, some of these cases had pneumonia. Most of the SARS-CoV-2 pneumonia published data came from hospitalised patients. This study set out to describe clinical characteristics of patients with SARS-CoV-2 pneumonia diagnosed in primary care across age groups and type of pneumonia. METHODS: Observational retrospective study obtaining clinical data from the electronic health records of patients who were followed-up by SARS-CoV-2 possible infection in a primary care practice in Madrid. All the cases were collected by in-person or remote consultation during the 10th March to the 7th of April. EXPOSURE: Diagnosis of SARS-CoV-2 pneumonia by chest X-ray ordered by the GP. Main outcomes and measures: Symptoms of SARS-CoV-2 pneumonia, physical examination and diagnostic tests as a blood test, nasopharyngeal swab results for RT-PCR (Reverse transcriptase-polymerase chain reaction) and chest X-ray results. RESULTS: The overall SARS-CoV-2 pneumonias collected were 172 (female 87 [50.6%], mean age 60.5 years standard deviation [SD] 17.0). Comorbidities were body mass index ≥ 25 kg/m2 (90 [52.3%]), hypertension (83 [48.3%]), dyslipidaemia (68 [39.5%]) and diabetes (33 [19.2%]). The sample was stratified by age groups (< 50 years, 50-75 years and ≥ 75 years). Clinical manifestations at onset were fever (144 [83.7%]), cough (140 [81.4%]), dyspnoea (103 [59.9%]) and gastrointestinal disturbances (72 [41.9%]). Day 7.8 (SD:4.1) from clinical onset was the mean day of pneumonia diagnosis. Bilateral pneumonia was more prevalent than unilateral (126 [73.3%] and 46 [26.7%]). Patients with unilateral pneumonia were prone to higher pulse oximetry (96% vs 94%, p < 0.001). We found differences between unilateral and bilateral cases in C-reactive protein (29.6 vs 81.5 mg/L, p < 0.001), and lymphocytes (1400.0 vs 1000.0E3/ml, p < 0.001). Complications were registered: 42 (100%) of patients ≥ 75 years were admitted into hospital; pulmonary embolism was only present at bilateral pneumonia (7 patients [5.6%]) and death occurred in 1 patient with unilateral pneumonia (2.2%) vs 10 patients (7.9%) with bilateral pneumonia ( p 0.170). CONCLUSION: Clinical manifestations of SARS-CoV-2 pneumonia were fever, cough and dyspnoea; this was especially clear in the elderly. We described different characteristics between unilateral and bilateral pneumonia.
Subject(s)
COVID-19 , Lung/diagnostic imaging , Pneumonia, Viral , Primary Health Care , SARS-CoV-2/isolation & purification , Symptom Assessment , Age Factors , COVID-19/diagnosis , COVID-19/epidemiology , COVID-19/physiopathology , COVID-19 Testing/methods , Causality , Electronic Health Records/statistics & numerical data , Female , Humans , Male , Middle Aged , Pneumonia, Viral/diagnosis , Pneumonia, Viral/epidemiology , Pneumonia, Viral/virology , Primary Health Care/methods , Primary Health Care/statistics & numerical data , Radiography, Thoracic/methods , Radiography, Thoracic/statistics & numerical data , Retrospective Studies , Spain/epidemiology , Symptom Assessment/methods , Symptom Assessment/statistics & numerical dataABSTRACT
Vaccination against retroviruses is a challenge because of their ability to stably integrate into the host genome, undergo long-term latency in a proportion of infected cells and thereby escape immune response. Since clearance of the virus is almost impossible once infection is established, the primary goal is to achieve sterilizing immunity. Besides efficacy, safety is the major issue since vaccination has been associated with increased infection or reversion to pathogenicity. In this review, we discuss the different issues that we faced during the development of an efficient vaccine against bovine leukemia virus (BLV). We summarize the historical failures of inactivated vaccines, the efficacy and safety of a live-attenuated vaccine and the economical constraints of further industrial development.
Subject(s)
Enzootic Bovine Leukosis/prevention & control , Leukemia Virus, Bovine/immunology , Viral Vaccines/immunology , Animals , Antibodies, Viral/immunology , Cattle , Vaccination/veterinary , Vaccines, Attenuated/immunologyABSTRACT
Foot and mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals, which causes severe economic losses in the livestock industry. Currently available vaccines are based on inactivated FMD virus (FMDV). Although inactivated virus vaccines have proved to be effective in FMD control, they have a number of disadvantages, including the need for high bio-containment production facilities and the lack of induction of immunological memory. Novel FMD vaccines based on the use of recombinant empty capsids have shown promising results. These recombinant empty capsids are attractive candidates because they avoid the use of virus in the production facilities but conserve its complete repertoire of conformational epitopes. However, many of these recombinant empty capsids require time-consuming procedures that are difficult to scale up. Achieving production of a novel and efficient FMD vaccine requires not only immunogenic antigens, but also industrially relevant processes. This review intends to summarize and compare the different strategies already published for the production of FMDV recombinant empty capsids, focusing on large-scale production.
Subject(s)
Capsid Proteins/genetics , Foot-and-Mouth Disease/drug therapy , Recombinant Proteins/genetics , Vaccines/genetics , Animals , Capsid/chemistry , Capsid/immunology , Capsid Proteins/immunology , Capsid Proteins/therapeutic use , Foot-and-Mouth Disease/immunology , Foot-and-Mouth Disease/virology , Foot-and-Mouth Disease Virus/drug effects , Foot-and-Mouth Disease Virus/immunology , Foot-and-Mouth Disease Virus/pathogenicity , Humans , Recombinant Proteins/immunology , Recombinant Proteins/therapeutic use , Vaccines/therapeutic use , Vaccines, Inactivated/genetics , Vaccines, Inactivated/immunologyABSTRACT
Bovine leukemia virus (BLV) is an important cattle pathogen that causes major economic losses worldwide, especially in dairy farms. The use of animal models provides valuable insight into the pathogenesis of viral infections. Experimental infections of sheep have been conducted using blood from BLV-infected cattle, infectious BLV molecular clones or tumor-derived cells. The Fetal Lamb Kidney cell line, persistently infected with BLV (FLK-BLV), is one of the most commonly used long-term culture available for the permanent production of virus. FLK-BLV cells or the viral particles obtained from the cell-free culture supernatant could be used as a source of provirus or virus to experimentally infect sheep. In this report, we aimed to determine the minimum amount of FLK-BLV cells or cell-free supernatant containing BLV needed to produce infection in sheep. We also evaluated the amount of antibodies obtained from a naturally-infected cow required to neutralize this infection. We observed that both sheep experimentally inoculated with 5000 FLK-BLV cells became infected, as well as one of the sheep receiving 500 FLK-BLV cells. None of the animals inoculated with 50 FLK-BLV cells showed evidence of infection. The cell-free FLK-BLV supernatant proved to be infective in sheep up to a 1:1000 dilution. Specific BLV antibodies showed neutralizing activity as none of the sheep became infected. Conversely, the animals receiving a BLV-negative serum showed signs of BLV infection. These results contribute to the optimization of a sheep bioassay which could be useful to further characterize BLV infection.
Subject(s)
Antibodies, Neutralizing/immunology , Antigen-Antibody Reactions , Antigens, Viral/immunology , Cells, Cultured/virology , Enzootic Bovine Leukosis/immunology , Leukemia Virus, Bovine/immunology , Animals , Cattle , Disease Models, Animal , Enzootic Bovine Leukosis/blood , Neutralization Tests , SheepABSTRACT
Vaccination has proved to be an efficient strategy to deal with viral infections in both human and animal species. However, protection of cattle against viral infections is still a major concern in veterinary science. During the last two decades, the development of efficient plant-based expression strategies for recombinant proteins prompted the application of this methodology for veterinary vaccine purposes. The main goals of viral bovine vaccines are to improve the health and welfare of cattle and increase the production of livestock, in a cost-effective manner. This review explores some of the more prominent recent advances in plant-made viral bovine vaccines against foot-and-mouth disease virus (FMDV), bovine rotavirus (BRV), bovine viral diarrhoea virus (BVDV), bluetongue virus (BTV) and bovine papillomavirus (BPV), some of which are considered to be the most important viral causative agents of economic loss in cattle production.
Subject(s)
Cattle Diseases/immunology , Cattle Diseases/virology , Viral Vaccines/immunology , Virus Diseases/veterinary , Animals , Cattle , Plants , Virus Diseases/immunology , Virus Diseases/virologyABSTRACT
We report the synthesis of hybrid supercapacitor electrodes by a novel reduction of GO with simultaneous incorporation of polyoxometalate. These hybrids show a 30% increase in specific capacitance and excellent stability after 10,000 cycles.
ABSTRACT
Bilateral lung transplantation is the treatment of end-stage lung diseases. However, sometimes a single lung transplant is performed. The technique is not exempt from complications such as acute hyperinflation of the native lung and changes in the diaphragm, predisposing to atelectasis and respiratory failure that can lead to negative results. Therefore, spontaneous breathing trials may fail and delay the weaning process. The combination of advanced monitoring tools, such as electrical impedance tomography and ultrasonography, to diagnose the cause of this failure, recognizing and quantifying the distribution of lung volume and its dynamic behavior could be crucial to improve outcomes. We present the case of a patient with a one-lung transplant and prolonged mechanical ventilation who, after presenting successive failures in the weaning process, underwent advanced monitoring in order to find the causes of the failure.
El trasplante de pulmón bilateral es el tratamiento de las enfermedades pulmonares en su etapa terminal. Sin embargo, a veces se realiza el trasplante de un solo pulmón. La técnica no está exenta de complicaciones como la hiperinsuflación aguda del pulmón nativo y cambios en el diafragma, predisponiendo a atelectasias e insuficiencia respiratoria que pueden derivar en resultados negativos. Por lo tanto, las pruebas de respiración espontánea pueden fallar y retrasar el proceso de desvinculación de la ventilación mecánica. La combinación de herramientas de monitorización avanzadas, como la tomografía por impedancia eléctrica y la ecografía, para diagnosticar la causa de este fallo, reconociendo y cuantificando la distribución del volumen pulmonar y su comportamiento dinámico, podría ser crucial para mejorar los resultados. Presentamos el caso de un paciente con trasplante unipulmonar y ventilación prolongada que falla en repetidas ocasiones durante la desvinculación de la ventilación mecánica, donde utilizamos herramientas de monitoreo avanzado para detectar la causa de la falla.
Subject(s)
Lung Transplantation , Respiration, Artificial , Humans , Ventilator Weaning/methods , Lung/diagnostic imaging , Tomography, X-Ray ComputedABSTRACT
Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis, an endemic disease in dairy cattle of Argentina. However, little is known about the seroprevalence of BLV in beef cattle. In this study, we conducted a cross-sectional study including farms from thirteen provinces of Argentina. A total of 5827 bovine serum samples were collected from 76 farms and analyzed using an in-house developed enzyme-linked immunosorbent assay. Information about herd management was collected through a questionnaire, and univariate and multivariate analyses were performed to detect risk factors associated with BLV infection. Herd-level seroprevalence was 71.05%, while the mean animal-level seroprevalence was 7.23% (median = 2.69%; min = 0, max = 75). Only two provinces had no positive BLV samples. The other eleven provinces showed more than 50% of their farms infected with BLV. The multivariate model revealed that BLV prevalence was significantly associated with the use of animals raised in the same farm for cattle replacement (P = 0.005), breeding cows by natural mating with a bull (P < 0.001), and weaning calves after 6 months of age (P = 0.011). This extensive study revealed that BLV seroprevalence in Argentine beef farms has increased during the last years and allowed identifying some management practices associated with BLV prevalence. These data deserve special attention because BLV infection in beef cattle seems to lead to a dissemination pattern similar to that observed during the last decades in dairy cattle, especially considering that Argentina is the sixth beef producer in the world, with about 5% of global beef production.
Subject(s)
Cattle Diseases , Enzootic Bovine Leukosis , Leukemia Virus, Bovine , Female , Cattle , Animals , Male , Seroepidemiologic Studies , Cross-Sectional Studies , Antibodies, Viral , Enzootic Bovine Leukosis/epidemiology , Risk Factors , Enzyme-Linked Immunosorbent Assay/veterinary , Cattle Diseases/epidemiologyABSTRACT
Hypoxia contributes to numerous pathophysiological conditions including inflammation-associated diseases. We characterized the impact of hypoxia on the immunometabolic cross-talk between cholesterol and interferon (IFN) responses. Specifically, hypoxia reduced cholesterol biosynthesis flux and provoked a compensatory activation of sterol regulatory element-binding protein 2 (SREBP2) in monocytes. Concomitantly, a broad range of interferon-stimulated genes (ISGs) increased under hypoxia in the absence of an inflammatory stimulus. While changes in cholesterol biosynthesis intermediates and SREBP2 activity did not contribute to hypoxic ISG induction, intracellular cholesterol distribution appeared critical to enhance hypoxic expression of chemokine ISGs. Importantly, hypoxia further boosted chemokine ISG expression in monocytes upon infection with severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2). Mechanistically, hypoxia sensitized toll-like receptor 4 (TLR4) signaling to activation by SARS-CoV-2 spike protein, which emerged as a major signaling hub to enhance chemokine ISG induction following SARS-CoV-2 infection of hypoxic monocytes. These data depict a hypoxia-regulated immunometabolic mechanism with implications for the development of systemic inflammatory responses in severe cases of coronavirus disease-2019 (COVID-19).
Subject(s)
COVID-19 , Interferons , Humans , Interferons/pharmacology , Monocytes , SARS-CoV-2 , Chemokines , Hypoxia , CholesterolABSTRACT
The risks and possible accidents related to the normal use of lithium-ion batteries remain a serious concern. In order to get a better understanding of thermal runaway (TR), the exothermic decomposition reactions in anode and cathode were studied, using a Simultaneous Thermal Analysis (STA)/Gas Chromatography-Mass Spectrometry (GC-MS)/Fourier Transform Infrared (FTIR) spectrometer system. These techniques allowed the identification of the reaction mechanisms in each electrode, owing to the analysis of evolved gaseous species, the amount of heat released and mass loss. These results provided insight into the thermal events happening within a broader temperature range than covered in previously published models. This allowed the formulation of an improved thermal model to depict TR. The heat of reaction, activation energy, and frequency factor (thermal triplets) for each major exothermic process at material level were investigated in a Lithium Nickel-Manganese-Cobalt-Oxide (NMC (111))-Graphite battery cell. The results were analyzed, and their kinetics were derived. These data can be used to successfully simulate the experimental heat flow.
ABSTRACT
OBJECTIVE: To explore the most frequent errors of the inhalation technique (main topic for the pharmacological treatment of exacerbations of asthma and/or chronic obstructive pulmonary disease) in the nursing staff of an emergency department, and to evaluate the efficacy of an educational intervention. METHODS: Prospective quasi-experimental study, carried out in a hospital in Argentina during 2019. For the evaluation of the nursing staff, the steps of the Separ-Alat Consensus were used. The educational intervention consisted of oral explanation and visual demonstration of the inhalation technique. Paired data analysis was performed, and absolute and relative frequencies were calculated with their respective 95% confidence intervals. RESULTS: A consecutive sample of 72 nurses was included, of which 95.83% (95%CI 88.30-99.13) made at least one error in the baseline assessment. The most frequent errors were the lack of indication to the patient to exhale prior to inhalation (83.72%), forgetting to remove the air chamber and repeat the steps before the second dose (81.40%). After immediate evaluation after receiving the educational intervention, 74.42% (95%CI 58.83-86.49) corrected the technique. However, at one month, 53.49% (23/43; 95%CI 37.66-68.86) performed the technique correctly. CONCLUSIONS: The educational intervention could be effective in correcting errors in this practice, although its effects tend to fade over time. Therefore, we consider periodic and systematic education relevant to maintain a correct inhalation technique among professionals.
Subject(s)
Nebulizers and Vaporizers , Pulmonary Disease, Chronic Obstructive , Administration, Inhalation , Emergency Service, Hospital , Humans , Prospective Studies , Pulmonary Disease, Chronic Obstructive/drug therapyABSTRACT
OBJECTIVE: To describe health care providers' knowledge about lung donation and donor lung management. METHODS: A descriptive, cross-sectional study based on an anonymous survey was conducted between March and September 2018 among health care professionals registered to Sociedad Argentina de Terapia Intensiva. RESULTS: Of the 736 respondents, the mean age was 40.5 years (standard deviation 8.9), and 61.3% were female. Sixty percent were physicians, 21.5% were nurses, and 17.9% were physiotherapists. Seventy-eight percent considered themselves appropriately informed about organ procurement, and 79.8% stated that they knew potential organ donor critical care management. The lung donor criteria were answered correctly by 71.3% of the respondents. However, after the donor's brain death, 51% made no changes to ventilator parameters, 22.9% were not aware of which parameters to reprogram, and 44.5% selected tidal volume of 6 - 8mL/kg and positive end expiratory pressure of 5cmH2O. For 85% of the health care providers, the type of apnea test chosen was disconnection from the ventilator, and only 18.5% used a lung management protocol. The most frequent interventions used in the case of arterial oxygen partial pressure/fractional inspired oxygen < 300 were positive end expiratory pressure titration, closed-circuit endotracheal suctioning, and recruitment maneuvers. CONCLUSION: Health care professionals surveyed in Argentina correctly answered most of the questions related to lung donor criteria. However, they lacked detailed knowledge about ventilatory settings, ventilatory strategies, and protocols for lung donors. Educational programs are key to optimizing multiorgan donation and should be focused on protecting the donor lungs to increase the numbers of organs available for transplantation.
OBJETIVO: Descrever o conhecimento de profissionais de saúde relacionado com doação de pulmão e manejo do doador. MÉTODOS: Realizou-se estudo descritivo, transversal, com base em levantamento anônimo conduzido entre março e setembro de 2019 envolvendo profissionais de saúde registrados na Sociedad Argentina de Terapia Intensiva. RESULTADOS: Dentre os 736 participantes, a média de idade foi de 30,5 anos (desvio-padrão de 8,9), sendo 61,3% do sexo feminino. Dentre os participantes, 60% eram médicos, 21,5% enfermeiros e 17,9% fisioterapeutas. Dentre os participantes, 68% se consideravam adequadamente informados com relação à procura de órgãos, e 79,8% afirmaram estar cientes do manejo de um potencial doador sob terapia intensiva. Os critérios relativos a um doador de pulmão foram respondidos corretamente por 71,3% dos participantes. Entretanto, após a morte cerebral do doador, 51% dos participantes não fariam modificações nos parâmetros de ventilação mecânica, 22,9% não sabiam quais parâmetros reprogramar, e 44,5% escolheriam um volume corrente de 6 - 8mL/kg e pressão positiva expiratória final de 5cmH2O. Para 85% dos profissionais de saúde, o tipo de teste de apneia escolhido foi desconexão do ventilador, e apenas 18,5% utilizariam um protocolo de controle. As intervenções mais frequentemente utilizadas no caso de pressão parcial de oxigênio/fração inspirada de oxigênio < 300 foram titulação da pressão positiva expiratória final, aspiração traqueal em circuito fechado e manobras de recrutamento. CONCLUSÃO: Os profissionais de saúde que participaram deste levantamento na Argentina responderam corretamente à maior parte das questões relacionadas aos critérios para o doador de pulmão. Contudo, faltou-lhes um conhecimento detalhado relativo aos parâmetros ventilatórios, às estratégias ventilatórias e aos protocolos para doadores de pulmão. Programas educacionais são fundamentais para otimizar a doação de múltiplos órgãos e devem focalizar a proteção dos pulmões do doador, com objetivo de incrementar o número de órgãos disponíveis para transplante.
Subject(s)
Tissue Donors , Tissue and Organ Procurement , Adult , Argentina , Cross-Sectional Studies , Female , Health Personnel , Humans , Lung , Male , Middle Aged , Surveys and QuestionnairesABSTRACT
BACKGROUND: Patients with COVID-19 are follow-up in primary care and long COVID is scarcely defined. The study aim was to describe SARS-CoV-2 pneumonia and cut-offs for defining long COVID in primary care follow-up patients. METHODS: A retrospective observational study in primary care in Madrid, Spain, was conducted. Data was collected during 6 months (April to September) in 2020, during COVID-19 first wave, from patients ≥ 18 years with SARS-CoV-2 pneumonia diagnosed. Variables: sociodemographic, comorbidities, COVID-19 symptoms and complications, laboratory test and chest X-ray. Descriptive statistics were used, mean (standard deviation (SD)) and medians (interquartile range (IQR)) respectively. Differences were detected applying X2 test, Student's T-test, ANOVA, Wilcoxon-Mann-Whitney or Kruskal-Wallis depending on variable characteristics. RESULTS: 155 patients presented pneumonia in day 7.8 from the onset (79.4% were hospitalized, median length of 7.0 days (IQR: 3.0, 13.0)). After discharge, the follow-up lasted 54.0 median days (IQR 42.0, 88.0) and 12.2 mean (SD 6.4) phone calls were registered per patient. The main symptoms and their duration were: cough (41.9%, 12 days), dyspnoea (31.0%, 15 days), asthenia (26.5%, 21 days). Different cut-off points were applied for long COVID and week 4 was considered the best milestone (28.3% of the sample still had symptoms after week 4) versus week 12 (8.3%). Patients who still had symptoms >4 weeks follow-up took place over 81.0 days (IQR: 50.5, 103.0), their symptoms were more prevalent and lasted longer than those ≤ 4 weeks: cough (63.6% 30 days), dyspnoea (54.6%, 46 days), and asthenia (56.8%, 29 days). Embolism was more frequent in patients who still had symptoms >4 weeks than those with symptoms ≤4 weeks (9.1% vs 1.8%, p value 0.034). CONCLUSION: Most patients with SARS-CoV-2 pneumonia recovered during the first 4 weeks from the beginning of the infection. The cut-off point to define long COVID, as persisting symptoms, should be between 4 to 12 weeks from the onset of the symptoms.
Subject(s)
COVID-19/complications , Primary Health Care/statistics & numerical data , Adult , COVID-19/epidemiology , Cities/epidemiology , Female , Humans , Male , Middle Aged , Retrospective Studies , Spain/epidemiology , Post-Acute COVID-19 SyndromeABSTRACT
Genomic heterogeneity and quasispecies composition of GB virus C (GBV-C) within plasma and lymphocyte subsets in a naturally infected blood donor were investigated. For this purpose, fragments from the 5' untranslated region (5' UTR) and the E2 gene recovered from plasma, B and T lymphocytes, were cloned and sequenced. A total of 63 clones was analysed: 95.2 % of them (n=60) - obtained from plasma and cells - were assigned to genotype 2b, while only three derived from plasma corresponded to genotyope 3. The G215A transition within this region was present in 90.9 % of the clones from B lymphocytes, but absent in the remaining cell compartments (P<0.01). Apparently, most of the circulating GBV-C quasispecies in this blood donor were related to the viral population infecting CD8(+) T cells, and B cells to a lesser extent. This is the first report showing the quasispecies nature of GBV-C in lymphocyte subsets within peripheral blood mononuclear cells.
Subject(s)
Flaviviridae Infections/virology , GB virus C/classification , GB virus C/genetics , Hepatitis, Viral, Human/virology , Lymphocyte Subsets/virology , Base Sequence , Blood Donors , Genetic Speciation , Genetic Variation , Humans , Molecular Sequence Data , Phylogeny , Viral ProteinsABSTRACT
Fetal bovine serum (FBS) used in cell culture may be contaminated with adventitious agents, which can affect the production of biologicals and the results of clinical laboratory tests. We carried out a retrospective study to determine the incidence of adventitious agent contamination of Argentinean irradiated FBS dating from 2015 to 2019. We analyzed FBS batches for mycoplasma and adventitious viruses (bovine pestiviruses, bovine adenovirus, bluetongue virus, bovine parainfluenza virus 3, rabies virus, bovine parvovirus, bovine herpesvirus 1, bovine respiratory syncytial virus, and reovirus). Cell passages followed by direct immunofluorescence were carried out to check viability of the mentioned adventitious agents. Also, molecular detection of mycoplasma and pestiviruses was performed on the FBS samples. The presence of neutralizing antibodies against pestiviruses was determined. Molecular analyses indicated that frequencies of mycoplasma and pestiviruses in FBS were 14% and 84%, respectively. All of the batches were seronegative for pestiviral antibodies. After cell passages, all FBS samples were negative for hemadsorbent agents and by immunofluorescence for all of the viral species analyzed; PCR assays were negative for mycoplasma and pestiviruses. Our results demonstrate that, of all adventitious agents tested, local FBS batches only had traces of mycoplasma and pestiviruses; gamma irradiation was effective in inactivating them.
Subject(s)
Mycoplasma/isolation & purification , Serum/microbiology , Serum/virology , Viruses/isolation & purification , Animals , Antibodies, Viral/isolation & purification , Cattle , Culture Media , Polymerase Chain Reaction/veterinary , Retrospective StudiesABSTRACT
Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals that causes severe economic losses in the livestock industry. Currently available vaccines are based on the inactivated FMD virus (FMDV). Although inactivated vaccines have been effective in controlling the disease, they have some disadvantages. Because of these disadvantages, investigations are being made to produce vaccines in low containment facilities. The use of recombinant empty capsids (also referred as Virus Like Particles, VLPs) has been reported to be a promising candidate as a subunit vaccine because it avoids the use of virus in the vaccine production and conserves the conformational epitopes of the virus. Mignaqui and collaborators have produced recombinant FMDV empty capsids from serotype A/ARG/2001 using a scalable technology in mammalian cells that elicited a protective immunity against viral challenge in a mouse model. However, further evaluation of the immune response elicited by these VLPs in cattle is required. In the present work we compare the effect that VLPs or inactivated FMDV has on bovine dendritic cells and the humoral response elicited in cattle after a single vaccination.
ABSTRACT
OBJECTIVE: To describe the results from the implementation of a respiratory care and mechanical ventilation protocol on potential lung donors who met the conditions for procurement. The secondary objective is to compare the results with historical data. METHODS: This was a retrospective, observational study. It included potential donors suitable for procurement of organs who had brain death and were hospitalized in critical care units of the Autonomous City of Buenos Aires from April 2017 to March 2018. Main variables: number of potential lung donors that reached the objective of procurement, rate of lungs procured, and rate of implanted lungs. Values of p < 0.05 were considered significant. RESULTS: Thirty potential lung donors were included, and 23 (88.5%; 95%CI 69.8 - 97.6) met the oxygenation objective. Twenty potential lung donors donated organs, of whom eight donated lungs, with which four double lung transplants and eight single lung transplants were performed. Seven of 12 lungs were procured and implanted in the preprotocol period, while all 12 were under the protocol (p = 0.38). The implantation rate was 58.3% (7/12) in the historical control period and 100% (12/12) (p = 0.04) in the study period.
OBJETIVO: Describir los resultados de la implementación de un protocolo de cuidados respiratorios y de ventilación mecánica en el potencial donante de pulmón, que cumplen las condiciones para ser procurados. El objetivo secundario es comparar los resultados con datos históricos. MÉTODOS: Estudio retrospectivo y observacional. Incluye potenciales donantes aptos para procuración de órganos con muerte encefálica internados en las áreas críticas de la Ciudad Autónoma de Buenos Aires, desde abril de 2017 hasta marzo de 2018. Variables principales: número de potencial donante de pulmón que alcanzan el objetivo de procuración, tasa de pulmones procurados y tasa de pulmones implantados. Se consideraron valores significativos p < 0,05. RESULTADOS: Se incluyeron 30 potenciales donantes de pulmón, 23 (88.5%; IC95% 69,8 - 97,6) cumplieron el objetivo de oxigenación. Veinte potenciales donantes de pulmón donaron órganos y de ellos, ocho donaron pulmones, con los cuales se realizaron 4 trasplantes bipulmonares y 8 unipulmonares. Los pulmones procurados e implantados en el periodo pre-protocolo fueron 7, mientras que durante el protocolo fueron 12 (valor p = 0,38). La tasa de implantación fue 58,3% (7/12) en el control histórico y 100% (12/12) (valor p = 0,04) en el periodo de estudio. CONCLUSIÓN: El protocolo permitió alcanzar el objetivo de oxigenación en la mayoría de los potenciales donantes de pulmón y una mejoría estadísticamente significativa en la tasa de implantación.
Subject(s)
Lung Transplantation , Tissue and Organ Procurement , Brain Death , Humans , Lung , Respiration, Artificial , Retrospective Studies , Tissue DonorsABSTRACT
The effect of charging and discharging lithium iron phosphate-graphite cells at different temperatures on their degradation is evaluated systematically. The degradation of the cells is assessed by using 10 charging and discharging temperature permutations ranging from -20 °C to 30 °C. This allows an analysis of the effect of charge and discharge temperatures on aging, and their associations. A total of 100 charge/discharge cycles were carried out. Every 25 cycles a reference cycle was performed to assess the reversible and irreversible capacity degradation. A multi-factor analysis of variance was used, and the experimental results were fitted showing: i) a quadratic relationship between the rate of degradation and the temperature of charge, ii) a linear relationship with the temperature of discharge, and iii) a correlation between the temperature of charge and discharge. It was found that the temperature combination for charging at +30 °C and discharging at -5 °C led to the highest rate of degradation. On the other hand, the cycling in a temperature range from -20 °C to 15 °C (with various combinations of temperatures of charge and discharge), led to a much lower degradation. Additionally, when the temperature of charge is 15 °C, it was found that the degradation rate is nondependent on the temperature of discharge.
Subject(s)
Graphite/chemistry , Iron/chemistry , Lithium/chemistry , Phosphates/chemistry , TemperatureABSTRACT
Foot-and-mouth disease (FMD) remains one of the most feared viral diseases affecting cloven-hoofed animals, and results in severe economic losses. Currently available vaccines are based on inactivated FMD virus (FMDV). The use of recombinant FMDV-like particles (VLPs) as subunit vaccines has gained importance because of their immunogenic properties and safety. We evaluated the production of FMD VLPs, via Agrobacterium-mediated transient expression, and the immunogenicity of these structures in mice. Leaves were infiltrated with pEAQ-HT and pRIC 3.0 vectors encoding the capsid precursor P1-2A and the protease 3C. The recombinant protein yield was 3-4 mg/kg of fresh leaf tissue. Both groups of mice immunized with purified VLPs and mice immunized with the crude leaf extract elicited a specific humoral response with similar antibody titers. Thus, minimally processed plant material containing transiently expressed FMD VLPs could be a scalable and cost-effective technology for the production of a recombinant subunit vaccine against FMDV.