ABSTRACT
We conducted a development and standardization of an IgG ELISA assay for serological detection of human orthohantavirus infections using the recombinant antigen rLECH13 produced in bacterial and derived from the LECHV. The evaluation and standardization were carried out by analyzing serum samples from a total of 50 patients with confirmed Hantavirus Pulmonary Syndrome (HPS) diagnosis through the reference technique, 50 negative sera, and 53 patients with other medical conditions. The data from the assay analysis showed a diagnostic sensitivity value of 95% and a diagnostic specificity of 80%. The high sensitivity of this novel assay leads us to conclude that rLECH13 is a feasible option for use in the immunodiagnostic of orthohantavirus infection. Additionally, it is crucial to have an antigen that can be produced under conditions that do not require highly complex laboratories. Furthermore, the new assay is cost-effective, reproducible, and demonstrates excellent performance.
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BACKGROUND: Chagas disease (CD), caused by the parasite Trypanosoma cruzi, is the most important endemic anthropozoonosis in Argentina. Since 2010, the World Health Organization has highlighted the urgent need to validate diagnostic systems that allow rapid detection of T. cruzi, infection in primary healthcare centers. Serological rapid diagnostic tests (RDTs) for T. cruzi, infection could be used to improve case management, as RDTs do not require specialized laboratories or highly trained staff to use them. We aimed to generate unbiased performance data of RDTs in Argentina, to evaluate their usefulness for improving T. cruzi, diagnosis rates. METHODS AND PRINCIPAL FINDINGS: This is a retrospective, laboratory-based, diagnostic evaluation study to estimate the clinical sensitivity/specificity of four commercially available RDTs for T. cruzi, using the Chagas disease diagnostic algorithm currently used in Argentina as the reference standard. In total, 400 serum samples were tested, 200 from individuals with chronic T. cruzi infection and 200 from individuals not infected with T. cruzi. All results were registered as the agreement of at least two operators who were blinded to the reference standard results. The sensitivity estimates ranged from 92.5-100% (95% confidence interval (CI) lower bound 87.9-98.2%); for specificity, the range was 76-96% (95% CI lower bound 69.5-92.3%). Most RDTs evaluated showed performances comparable with the reference standard method, showing almost perfect concordance (Kappa 0.76-0.92). CONCLUSIONS: Our study demonstrates that, under controlled laboratory conditions, commercially available RDTs for CD have a performance comparable to the Argentinian diagnostic algorithm, which is based on laboratory-based serological tests. For the next stage of our work, the RDTs will be evaluated in real-world settings.
Subject(s)
Chagas Disease , Trypanosoma cruzi , Humans , Argentina/epidemiology , Retrospective Studies , Urban Population , Rapid Diagnostic Tests , Chagas Disease/diagnosis , Chagas Disease/epidemiology , Antibodies , Sensitivity and Specificity , Antibodies, ProtozoanABSTRACT
Bolivia has the highest incidence of Chagas disease (CD) worldwide. Caused by the parasite Trypanasoma cruzi, CD is generally a chronic condition. Diagnosis is logistically and financially challenging, requiring at least two different laboratory-based serological tests. Many CD cases are missed; in Bolivia it is estimated just 6% of individuals chronically infected with T. cruzi get diagnosed. Achieving control on the way to elimination of CD requires a radical simplification of the current CD testing pathways, to overcome the barriers to accessing CD treatment. We aimed to generate unbiased performance data of lateral flow assays (LFAs) for T. cruzi infection in Bolivia, to evaluate their usefulness for improving T. cruzi diagnosis rates in a precise and efficient manner. This retrospective, laboratory-based, diagnostic evaluation study sought to estimate the sensitivity/specificity of 10 commercially available LFAs for T. cruzi, using the current CD diagnostic algorithm employed in Bolivia as the reference test method. All tests were blinded at the study site and performed by three operators. In total, 470 serum samples were tested, including 221 and 249 characterized as CD-positive/-negative, respectively. The LFAs were scored according to their relative importance using a decision-tree-based algorithm, with the mean decrease in Gini index as the scoring metric. The estimates of sensitivities ranged from 62.2-97.7% (95% confidence interval (CI) lower bound 55.0-94.7%); for specificities the range was 78.6-100% (95% CI lower bound 72.0-97.5%); 5/10 and 6/10 tests had sensitivity >90% and specificity >95%, respectively. Four LFAs showed high values of both sensitivity (93-95%) and specificity (97-99%). The agreement between 6 LFAs and the reference tests was almost perfect (Kappa 0.83-0.94). Most LFAs evaluated thus showed performances comparable with current laboratory-based diagnostic methods.
Subject(s)
Chagas Disease , Trypanosoma cruzi , Humans , Bolivia , Retrospective Studies , Chagas Disease/parasitology , Sensitivity and SpecificityABSTRACT
Trichinellosis is a foodborne disease caused by ingestion of raw or undercooked meat containing Trichinella spp. larvae. Consumption of wild boar (Sus scrofa) meat represents an important source of human trichinellosis worldwide. In El Palmar National Park (EPNP), Argentina, invasive alien wild boars are controlled and meat from culled animals is released for public consumption following on-site artificial digestion (AD) testing. Meat trimmings and offal from the control program are often used as food for dogs (Canis familiaris). We evaluated infection and exposure to Trichinella spp. in wild boars from EPNP, as well as exposure to Trichinella spp. and associated risk factors in dogs and human consumers of wild boar meat. Trichinella spp. larvae were detected in muscle samples from 5/49 wild boars by AD (10.2%; 95% confidence interval [CI], 3.8%-23%), with a mean burden of 0.24 larvae per gram (lpg; range, 0.06-0.95 lpg). Anti-Trichinella antibodies were not detected in wild boar serum samples (n=42). In dogs, 12/34 were seropositive to Trichinella spp. (35.29%; 95%, CI, 20.3%-53.5%). Immunoglobulin (Ig) G antibodies were not detected in human serum samples (n=63). Our results reveal the presence, albeit at low prevalence, of Trichinella spp. in wild boars and exposure in dogs fed game offal. These findings suggest that the low prevalence and parasitic load in wild boars, together with the best practices applied by EPNP culling program personnel, contribute to keeping the risk of infection in people low. The dog results highlight that the parasite is circulating in the area, and therefore the risk of infection is not negligible. We recommend the implementation of an animal surveillance strategy in order to monitor the evolution of this zoonosis in the study area.
Subject(s)
Dog Diseases , Nitrophenols , Swine Diseases , Trichinella , Trichinellosis , Swine , Humans , Animals , Dogs , Trichinellosis/epidemiology , Trichinellosis/veterinary , Trichinellosis/parasitology , Argentina/epidemiology , Parks, Recreational , Swine Diseases/epidemiology , Swine Diseases/parasitology , Meat/parasitology , Immunoglobulin G , Sus scrofa , Dog Diseases/epidemiology , Epoxy CompoundsABSTRACT
INTRODUCTION: A comprehensive pathophysiological mechanism to explain the relationship between high-salt intake and hypertension remains undefined. Evidence suggests that chloride, as the accompanying anion of sodium in dietary salt, is necessary to develop hypertension. We evaluated whether reducing dietary Cl- while keeping a standard Na+ intake modified blood pressure, cardiac hypertrophy, renal function, and vascular contractility after angiotensin II (AngII) infusion. METHODS: C56BL/6J mice fed with standard Cl- diet or a low-Cl- diet (equimolar substitution of Cl- by a mixture of Na+ salts, both diets with standard Na+ content) received AngII (infusion of 1.5 mg/kg/day) or vehicle for 14 days. We measured systolic blood pressure (SBP), glomerular filtration rate (GFR), natriuretic response to acute saline load, and contractility of aortic rings from mice infused with vehicle and AngII, in standard and low-Cl- diet. RESULTS: The mice fed the standard diet presented increased SBP and cardiac hypertrophy after AngII infusion. In contrast, low-Cl- diet prevented the increase of SBP and cardiac hypertrophy. AngII-infused mice fed a standard diet presented hampered natriuretic response to saline load, meanwhile the low-Cl- diet preserved natriuretic response in AngII-infused mice, without change in GFR. Aortic rings from mice fed with standard diet or low-Cl- diet and infused with AngII presented a similar contractile response. CONCLUSION: We conclude that the reduction in dietary Cl- as the accompanying anion of sodium in salt is protective from AngII pro-hypertensive actions due to a beneficial effect on kidney function and preserved natriuresis.
Subject(s)
Angiotensin II , Blood Pressure , Hypertension , Kidney , Animals , Mice , Angiotensin II/pharmacology , Blood Pressure/drug effects , Cardiomegaly/prevention & control , Cardiomegaly/chemically induced , Chlorides/administration & dosage , Chlorides/pharmacology , Glomerular Filtration Rate/drug effects , Hypertension/chemically induced , Hypertension/prevention & control , Kidney/drug effects , Mice, Inbred C57BL , Sodium Chloride, Dietary/adverse effects , Sodium Chloride, Dietary/administration & dosageABSTRACT
Campylobacter fetus is an important veterinary pathogen that causes campylobacteriosis. This disease causes decreased productivity of cattle by inducing reproductive losses. Although several virulence factors have been recognized in C. fetus, including a cytolethal distending toxin (CDT), the exact mechanism responsible for embryonic death remains unknown. The aim of this work was to evaluate the effect of C. fetus subsp. fetus (Cff) and C. fetus subsp. venerealis (Cfv), and their toxin activity on the in vitro fertilization of bovine ova and early embryonic development. Two different experiments were performed. In Experiment 1, a total of 1524 cumulus-oocyte complexes (COCs) were inseminated, distributed into three groups: two of them infected with the microorganism (Cff, Cfv) and a control group. Percentages COCs cleaved were similar among groups (p = 0.1243); however, the embryonic development rate (blastocyst at day 7) in the control group was greater (p < 0.001) than those obtained in Cff and Cfv groups. In experiment 2, a total of 746 COCs were inseminated, divided into three groups: two of them treated with the bacterial-free culture filtrates to test toxin activity (Cff-CDT, Cfv-CDT) and a control group. Both cleavage and embryonic development rates were greater (p < 0.001) in the control group than those obtained in Cff-CDT and Cfv-CDT groups. This study provides evidence that both subspecies of C. fetus do not interfere with fertilization but do affect in vitro embryonic development. It is the first report on the biological effect of the CDT on bovine embryonic development.
Subject(s)
Campylobacter Infections , Cattle Diseases , Pregnancy , Female , Cattle , Animals , Campylobacter fetus , Campylobacter Infections/veterinary , Fertilization in Vitro/veterinary , Blastocyst , Embryonic DevelopmentABSTRACT
Objetivo: Describir hallazgos clínicos, quirúrgicos e imagenológicos de la patología anorrectal en pacientes con Viruela Símica. Materiales y Métodos: Reporte de serie de casos clínicos, observacional, en pacientes con Viruela Símica tratados en un centro de salud privado desde agosto 2022 a diciembre 2022. Resultados: El 100% de los casos pertenecía al sexo masculino, la edad promedio fue de 32 años, todos los pacientes mantuvieron relaciones sexuales con otros hombres, la mayoría debuto con síntomas rectales, incluyendo proctalgia, prurito anal y heces diarreicas con mucosidad. El diagnóstico se confirmó mediante PCR (reacción de polimerasa en cadena) y el tratamiento fue sintomático, con mejoría en todos los pacientes en un seguimiento de 10 días. Discusión: La Viruela Símica es una enfermedad viral que se encuentra, principalmente, en regiones remotas de África Central, con brotes esporádicos en países africanos. Está relacionada con el virus de la viruela y se transmite, principalmente, del contacto con animales infectados o fluidos corporales. La enfermedad presenta síntomas similares a la gripe, seguidos del desarrollo de una erupción cutánea que progresa a pústulas y costras. No hay tratamiento específico para la viruela símica, el manejo medico alivia los síntomas. El brote de viruela símica en 2022 presentó características novedosas y atípicas en su forma de presentación. Conclusión: Los síntomas rectales asociados a una historia clínica compatible con viruela símica se deben considerar como diagnóstico diferencial a la proctitis por esta etiología específica y emergente.
Objective: To describe clinical, surgical and imaging findings of anorectal pathology in patients with monkeypox. Materials and Methods: Observational clinical case series report in patients with monkeypox treated in a private health center from August 2022 to December 2022. Results: 100% of the cases were male, the average age was 32 years old, all patients had sexual intercourse with other men, most debuted with rectal symptoms, including proctalgia, anal itching, and diarrheal stools with mucus. Diagnosis was confirmed by PCR (polymerase chain reaction) and treatment was symptomatic, with improvement in all patients at a 10-day follow-up. Discussion: Monkeypox is a viral disease found mainly in remote regions of Central Africa, with sporadic outbreaks in African countries. It is related to the smallpox virus and is transmitted primarily from contact with infected animals or bodily fluids. The disease presents with flu-like symptoms, followed by the development of a skin rash that progresses to pustules and scabs. There is no specific treatment for monkeypox; medical management alleviates the symptoms. The monkeypox outbreak in 2022 presented novel and atypical characteristics in its presentation. Conclusion: Rectal symptoms associated with a clinical history compatible with monkeypox should be considered as a differential diagnosis of proctitis due to this specific and emerging etiology.
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OBJECTIVES: Shiga toxin-producing Escherichia coli strains LAA-positive are important cause of human infection. The capability to adhere to epithelial cells is a key virulence trait, and genes codified in LAA pathogenicity island could be involved in the adhesion during the pathogenesis of LAA-positive STEC strains. Thus, our objectives were to compare hes-negative and hes-positive STEC strains in their adherence capability to epithelial cells (HEp-2) and to evaluate the expression levels of the hes, iha, and tpsA in the bacteria adhered and non-adhered to HEp-2 cells. These genes are encoded in LAA, and are virulence factors that participate in adhesion and autoaggregation. RESULTS: We could not observe differences between the adhesion of strains but also in the expression level of of hes, iha, and tpsA. Genes encoded in LAA contribute to the adhesion phenotype though the expression of STEC adhesins is a coordinated event that depends not only the strain but also on the environment as well as its genetic background. Therefore, the results of this study suggest that LAA ,the most prevalent PAI among LEE-negative STEC strains, plays a role in pathogenesis.
Subject(s)
Escherichia coli Proteins , Shiga-Toxigenic Escherichia coli , Humans , Adhesins, Bacterial/genetics , Epithelial Cells/microbiology , Escherichia coli Infections/genetics , Escherichia coli Infections/microbiology , Escherichia coli Proteins/genetics , Shiga-Toxigenic Escherichia coli/genetics , Shiga-Toxigenic Escherichia coli/metabolism , Virulence Factors/genetics , Virulence Factors/metabolism , Cell LineABSTRACT
Abstract Leptospirosis is a neglected zoonosis that is widely distributed in the world. Although it is endemic in Argentina, prevalence remains unknown. The aims of the study were: (i) to determine the prevalence of leptospirosis in humans from a rural community in Tandil Argentina, (ii) to identify infecting Leptospira spp. serogroups, (iii) to identify factors associated with the infection, (iv) to estimate the population attributable fraction (PAF) of the risk factors and (v) to determine the spatial patterns of disease presentation and related risk factors. Blood samples from 202 participants were collected. A survey was conducted to obtain clinical and epidemiological data. Serological testing was performed by the microscopic agglutination test (MAT). Univariate and multivariate methods were applied to evaluate associations. Spatial clusters were investigated for seroprevalence and risk factors. Antibodies were found in 32.2% of participants (95% CI: 25.8-39.1). The most prevalent serogroup was Hebdomadis followed by Sejroe; Icterohaemorrhagiae; Tarassovi and Canicola. Living at lower altitudes (OR: 13.04; 95% CI: 2.60-65.32); not having access to water supply network (OR: 2.95; 95% CI: 1.30-6.69); living close to flooded streets (OR: 2.94; 95% CI: 1.14-7.69) and practicing water sports (OR: 3.12; 95% CI: 1.12-8.33) were associated with seropositivity. Factors related with housing characteristics, services and infrastructure had the higher PAF (from 17% to 81%). A spatial cluster with higher rates of positivity and of the main risk factors was determined. This work contributes useful data for specific preventive measures that should be implemented for the control of the disease.
Resumen La leptospirosis es una enfermedad desatendida, ampliamente distribuida a nivel mundial. Aunque es endémica en Argentina, su prevalencia es desconocida. Los objetivos de este estudio fueron los siguientes: (i) determinar la prevalencia de leptospirosis humana en comunidades rurales del partido de Tandil (Argentina), (ii) identificar serogrupos infectantes de Leptospira spp., (iii) identificar factores de riesgo asociados, (iv) estimar la fracción atribuible poblacional (FAP) de los factores de riesgo y (v) determinar los patrones espaciales de la enfermedad y de los factores de riesgo. Se tomaron muestras de sangre a 202 personas, y se registró información clínica y epidemiológica. El diagnóstico se realizó por microaglutinación (MAT). Para evaluar asociaciones, se utilizaron métodos univariados y multivariados. Se estudiaron clusters espaciales de la seroprevalencia y de los factores de riesgo. El 32,2% de los participantes (IC 95%: 25,8-39,1) presentaron anticuerpos. Los serogrupos más prevalentes fueron Hebdomadis, Sejroe, Icterohaemorrhagiae, Tarassovi y Canicola. Vivir a menores altitudes (OR: 13,04; IC 95%: 2,60-65,32) y cerca de calles inundables (OR: 2,94; IC 95%: 1,14-7,69), la falta de acceso a agua de red (OR: 2,95; IC 95%: 1,30-6,69) y la práctica de deportes acuáticos (OR: 3,12; IC 95%: 1,12-8,33) estuvieron asociados con la seropositividad. Factores relacionados con las características de las viviendas, los servicios y la infraestructura tuvieron mayor proporción de FAP (17 al 81%). Se encontró un área de mayor riesgo de presentación de individuos seropositivos y de los principales factores de riesgo. Este trabajo provee información útil para generar medidas preventivas específicas que podrían ser aplicadas para controlar esta enfermedad.
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The SARS-CoV2 promotes dysregulation of Renin-Angiotensin-Aldosterone. The result is excessive retention of water, producing a state of noxious hypervolemia. Consequently, in COVID-19 injury lung is pulmonary edema. Our report is a case-control study, retrospective. We included 116 patients with moderate-severe COVID-19 lung injury. A total of 58 patients received standard care (Control group). A total of 58 patients received a standard treatment with a more negative fluid balance (NEGBAL group), consisting of hydric restriction and diuretics. Analyzing the mortality of the population studied, it was observed that the NEGBAL group had lower mortality than the Control group, p = 0.001. Compared with Controls, the NEGBAL group had significantly fewer days of hospital stay (p < 0.001), fewer days of ICU stay (p < 0.001), and fewer days of IMV (p < 0.001). The regressive analysis between PaO2/FiO2BAL and NEGBAL demonstrated correlation (p = 0.04). Compared with Controls, the NEGBAL group showed significant progressive improvement in PaO2/FiO2 (p < 0.001), CT score (p < 0.001). The multivariate model, the vaccination variables, and linear trends resulted in p = 0.671 and quadratic trends p = 0.723, whilst the accumulated fluid balance is p < 0.001. Although the study has limitations, the promising results encourage more research on this different therapeutic approach, since in our research it decreases mortality.
ABSTRACT
Leptospirosis is a neglected zoonosis that is widely distributed in the world. Although it is endemic in Argentina, prevalence remains unknown. The aims of the study were: (i) to determine the prevalence of leptospirosis in humans from a rural community in Tandil Argentina, (ii) to identify infecting Leptospira spp. serogroups, (iii) to identify factors associated with the infection, (iv) to estimate the population attributable fraction (PAF) of the risk factors and (v) to determine the spatial patterns of disease presentation and related risk factors. Blood samples from 202 participants were collected. A survey was conducted to obtain clinical and epidemiological data. Serological testing was performed by the microscopic agglutination test (MAT). Univariate and multivariate methods were applied to evaluate associations. Spatial clusters were investigated for seroprevalence and risk factors. Antibodies were found in 32.2% of participants (95% CI: 25.8-39.1). The most prevalent serogroup was Hebdomadis followed by Sejroe; Icterohaemorrhagiae; Tarassovi and Canicola. Living at lower altitudes (OR: 13.04; 95% CI: 2.60-65.32); not having access to water supply network (OR: 2.95; 95% CI: 1.30-6.69); living close to flooded streets (OR: 2.94; 95% CI: 1.14-7.69) and practicing water sports (OR: 3.12; 95% CI: 1.12-8.33) were associated with seropositivity. Factors related with housing characteristics, services and infrastructure had the higher PAF (from 17% to 81%). A spatial cluster with higher rates of positivity and of the main risk factors was determined. This work contributes useful data for specific preventive measures that should be implemented for the control of the disease.
Subject(s)
Leptospira , Leptospirosis , Humans , Rural Population , Seroepidemiologic Studies , Argentina/epidemiology , Leptospirosis/epidemiology , Antibodies, Bacterial , Risk Factors , Spatial AnalysisABSTRACT
Acyline contraception has been described in cats, but few data are available on the drug's long-term effect on growth. The relevant data cover until puberty with no radiographic description. We investigated the radiographic parameters throughout bone growth in order to more completely determine the drug's safety. Thirteen male and 12 female cats were studied, with the kittens being randomly assigned to one of the following groups within the first 24 hours of birth: ACY, subcutaneous acyline, 33 µg/100 g, which injection was repeated weekly until age 3 months; or CO, untreated control. Body measurements were recorded weekly and radiographic parameters obtained from monthly radiographs of the antebrachium. In the ACY and CO male and female kittens, the body weight, withers height, and body length plus the age at the end of body growth and radial growth remained similar throughout the study (P> .05). Both female groups finished radial growth before the males (P< .05). The ACY females evidenced a longer radial length between the eighth and 28th weeks (P< .05). All groups closed their proximal and distal physes within the normal ranges described for the species. The bone-cortex width was lower in the ACY vs. the CO animals at weeks 52 and 60 in the males and at weeks 24, 48, 52, and 56 in the females (P< .05) The transient greater radial length and lower bone-cortex thickness observed in the treated cats were compensated for at the end of growth with no adverse clinical effects being observed. In conclusion, acyline as a contraceptive did not evidence a permanent or severe effect on domestic-cat growth.
Subject(s)
Gonadotropin-Releasing Hormone , Oligopeptides , Animals , Cats , Contraceptive Agents , Female , Hormone Antagonists , Male , Oligopeptides/adverse effectsABSTRACT
AIM: To analyze the presence of fetal myocardial dysfunction in intrahepatic cholestasis of pregnancy (ICP) at diagnosis. METHODS: This prospective cohort study included 49 pregnant participants with ICP at diagnosis and 49 nonaffected controls from a single public hospital. ICP was diagnosed based on clinical symptoms after excluding other causes of pruritus and presence of autoimmune diseases. Total bile acids were not obtained in this cohort. ICP pregnancies were assessed with a functional echocardiography at diagnosis including PR-interval, isovolumetric contraction time (ICT), ejection time (ET), and isovolumetric relaxation time (IRT) for electrical, systolic, and diastolic function, respectively. Controls were assessed at recruitment. Perinatal outcomes were obtained from delivery books. The main outcome was the presence of PR-interval prolongation or first-degree fetal heart block, and echographic signs of diastolic and systolic dysfunction. RESULTS: Compared to controls, ICP were above upper limit in conjugated bilirubin (2.0% vs. 20.4%; p = 0.008), aspartate aminotransferase (2.0% vs. 24.5%; p = 0.002), and alanine aminotransferase (4.1% vs. 28.6%; p = 0.002). ICP was associated with a higher PR-interval (130 ± 12 ms vs. 121 ± 6 ms; p < 0.0001) with five first-degree fetal heart blocks. IRT was significantly higher in ICP (42 ± 6 ms vs. 37 ± 5 ms; p = 0.0001), with no differences in ICT and ET. PR-interval trend was only positively correlated with IRT in ICP pregnancies (p = 0.04 and p = 0.34, in ICP and controls, respectively). CONCLUSIONS: Our study demonstrates that fetuses affected by maternal ICP are associated with electrical and diastolic myocardial dysfunction. More studies focused on antenatal and postnatal functional echocardiography are necessary to validate our results and consider these markers in the clinical management of ICP pregnancies.
Subject(s)
Cholestasis, Intrahepatic , Heart Diseases , Pregnancy Complications , Bile Acids and Salts , Cholestasis, Intrahepatic/complications , Cholestasis, Intrahepatic/diagnostic imaging , Cohort Studies , Female , Fetus , Humans , Pregnancy , Pregnancy Complications/diagnosis , Prospective StudiesABSTRACT
Brucella ovis is an economically important cause of epididymitis in rams worldwide. Polymeric BLSOmp31 was previously identified as a protective immunogen against this pathogen. In this study, BLSOmp31 was formulated with a modified version of ISCOMATRIX adjuvant called ISPA (BLSOmp31/ISPA) and was administered in BALB/C by the subcutaneous and ocular route. The systemic and mucosal immune responses, the opsonic activity of antibodies and the protection conferred against B. ovis were evaluated. BLSOmp31+ISPA injected subcutaneously or by ocular route induced significantly higher IgG antibody levels with a mixed Th1/Th2 profile compared to non-immunized mice. IgA and IgG were detected in sera and nasal, tracheobronchial, vaginal secretions, tears and faeces, from SC immunized mice while in the group immunized by the ocular route a slight increase in both isotypes was mainly observed in all secretions, except in vaginal fluid. Opsonic antibodies stimulated binding and increased uptake of PHrodo™ Green-labelled B. ovis by neutrophils and monocytes. BLSOmp31 administered subcutaneously induced the highest levels of IFN-É£. The ocular immunization not only produced significant levels of this cytokine but also IL-4 compared to non-immunized mice. Both, subcutaneous and ocular routes of immunization, significantly protected against B. ovis infection. These results indicate that BLSOmp31/ISPA administered parenterally or by ocular route is a safe and effective vaccine against B. ovis in the murine model.
Subject(s)
Brucella ovis , Brucellosis , Rodent Diseases , Animals , Antibodies, Bacterial , Antigens, Bacterial , Brucellosis/prevention & control , Brucellosis/veterinary , Female , Male , Mice , Mice, Inbred BALB C , SheepABSTRACT
Retrograde ejaculation (RE) is a condition defined as the backward flow of the semen during ejaculation, and when present can result in male infertility. RE may be partial or complete, resulting in either low seminal volume or complete absence of the ejaculate (dry ejaculate). RE can result from anatomic, neurological or pharmacological conditions. The treatment approaches outlined are determined by the cause. Alkalinizing urinary pH with oral medications or by adding sperm wash media into the bladder prior to ejaculation may preserve the viability of the sperm. This article provides a step-by-step guide to diagnose RE and the optimal techniques to retrieve sperm.
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In COVID-19, pulmonary edema has been attributed to "cytokine storm". However, it is known that SARS-CoV2 promotes angiotensin-converting enzyme 2 deficit, increases angiotensin II, and this triggers volume overload. Our report is based on COVID-19 patients with tomographic evidence of pulmonary edema and volume overload to whom established a standard treatment with diuretic (furosemide) guided by objectives: Negative Fluid Balance (NEGBAL approach). Retrospective observational study. We reviewed data from medical records: demographic, clinical, laboratory, blood gas, and chest tomography (CT) before and while undergoing NEGBAL, from 20 critically ill patients. Once the NEGBAL strategy was started, no patient required mechanical ventilation. All cases reverted to respiratory failure with NEGBAL, but subsequently two patients died from sepsis and acute myocardial infarction (AMI). The regressive analysis between PaO2/FiO2BAL and NEGBAL demonstrated correlation (p < 0.032). The results comparing the Pao2Fio2 between admission to NEGBAL to NEGBAL day 4, were statistically significant (p < 0.001). We noted between admission to NEGBAL and day 4 improvement in CT score (p < 0.001), decrease in the superior vena cava diameter (p < 0.001) and the decrease of cardiac axis (p < 0.001). Though our study has several limitations, we believe the promising results encourage further investigation of this different pathophysiological approach.
ABSTRACT
BACKGROUND: Current algorithm for Congenital Chagas Disease (cCD) diagnosis is unsatisfactory due to low sensitivity of the parasitological methods. Moreover, loss to follow-up precludes final serodiagnosis after nine months of life in many cases. A duplex TaqMan qPCR kit for Trypanosoma cruzi DNA amplification was prospectively evaluated in umbilical cord (UCB) and peripheral venous blood (PVB) of infants born to CD mothers at endemic and non-endemic sites of Argentina. METHODS: We enrolled and followed-up 370 infants; qPCR was compared to gold-standard cCD diagnosis following studies of diagnostic accuracy guidelines. FINDINGS: Fourteen infants (3·78%) had cCD. The qPCR sensitivity and specificity were higher in PVB (72·73%, 99·15% respectively) than in UCB (66·67%, 96·3%). Positive and negative predictive values were 80 and 98·73% and 50 and 98·11% for PVB and UCB, respectively. The Areas under the Curve (AUC) of ROC analysis for qPCR and micromethod (MM) were 0·81 and 0·67 in UCB and 0·86 and 0·68 in PVB, respectively. Parasitic loads ranged from 37·5 to 23,709 parasite equivalents/mL. Discrete typing Unit Tc V was identified in five cCD patients and in six other cCD cases no distinction among Tc II, Tc V or Tc VI was achieved. INTERPRETATION: This first prospective field study demonstrated that qPCR was more sensitive than MM for early cCD detection and more accurate in PVB than in UCB. Its use, as an auxiliary diagnostic tool to MM will provide more accurate records on cCD incidence. FUNDING: FITS SALUD 001-CHAGAS (FONARSEC, MINCyT, Argentina) to the Public-Private Consortium (INGEBI-CONICET, INP-ANLIS MALBRAN and Wiener Laboratories); ERANET-LAC-HD 328 to AGS and PICT 2015-0074 (FONCYT, MinCyT) to AGS and FA.
Subject(s)
Chagas Disease/diagnosis , Real-Time Polymerase Chain Reaction/methods , Adult , Chagas Disease/congenital , Early Diagnosis , Female , Humans , Infant, Newborn , Male , Reagent Kits, Diagnostic/standards , Real-Time Polymerase Chain Reaction/standards , Sensitivity and SpecificityABSTRACT
Our aim was to evaluate the analytical and clinical performance of the SARS-CoV-2 molecular detection kits used in Argentina. Nine real-time reverse-transcription polymerase chain reaction (RT-qPCR) and three reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assays were evaluated using the World Health Organization (WHO) recommended test as reference method. A secondary standard calibrated for the E, N and RdRp genes against the Pan American Health Organization-World Health Organization-International Standard was used to calculate the limit of detection (LoD). A panel of artificial clinical samples, 32 positive and 30 negative for SARS-CoV-2, were analyzed to estimate the kappa concordance (κ) and the diagnostic performance. Differences among the LoD values for the target genes amplified by each kit were >1 log copies/reaction. The κ for the RT-qPCR kits was greater than 0.9, whereas that for the RT-LAMP assays ranged from 0.75 to 0.93. The clinical performance of RT-qPCR kits showed 100% specificity and high sensitivity, although with variations according to the gene analyzed. The E and N genes provided greater clinical sensitivity, whereas the RdRp gene increased the clinical specificity. The RT-LAMP assays revealed a variable diagnostic performance. The information provided can be useful to choose the most appropriate diagnostic test and may contribute to the establishment of a consensus in the diagnosis of SARS-CoV-2 in Argentina and the region.
Subject(s)
COVID-19 Nucleic Acid Testing/methods , Molecular Diagnostic Techniques/methods , Nucleic Acid Amplification Techniques/methods , Real-Time Polymerase Chain Reaction/methods , Argentina , Calibration , Humans , Limit of Detection , SARS-CoV-2/geneticsABSTRACT
Hepatocellular carcinoma (HCC) arises in the setting of advanced liver fibrosis, a dynamic and complex inflammatory disease. The tumor microenvironment (TME) is a mixture of cellular components including cancer cells, cancer stem cells (CSCs), tumor-associated macrophages (TAM), and dendritic cells (DCs), which might drive to tumor progression and resistance to therapies. In this work, we study the effects of 4-methylumbelliferone (4Mu) on TME and how this change could be exploited to promote a potent immune response against HCC. First, we observed that 4Mu therapy induced a switch of hepatic macrophages (MÏ) towards an M1 type profile, and HCC cells (Hepa129 cells) exposed to conditioned medium (CM) derived from MÏ treated with 4Mu showed reduced expression of several CSCs markers and aggressiveness. HCC cells incubated with CM derived from MÏ treated with 4Mu grew in immunosuppressed mice while presented delayed tumor progression in immunocompetent mice. HCC cells treated with 4Mu were more susceptible to phagocytosis by DCs, and when DCs were pulsed with HCC cells previously treated with 4Mu displayed a potent antitumoral effect in therapeutic vaccination protocols. In conclusion, 4Mu has the ability to modulate TME into a less hostile milieu and to potentiate immunotherapeutic strategies against HCC.