Characterization of biomarkers in blood serum for cancer diagnosis in dogs using Raman spectroscopy.

Biomarkers of cancer in sera of domestic dogs were detected through Raman spectroscopy with 830 nm excitation. Raman spectra of sera from 61 dogs (31 healthy and 30 with cancer, resulting in 154 and 200 spectra, respectively) were submitted to principal component analysis (PCA) for feature extraction and partial least squares (PLS) regression for discrimination between Healthy and Cancer groups. In the PCA, the peaks at 1132, 1342, 1368, and 1453 cm-1 (albumin and phenylalanine) were higher for the Cancer group. The "redshift" of the peaks at 621, 1003, and 1032 cm-1 (conformational change in proteins and/or bonds at sites close to the aromatic ring of amino acids) occurred in the Cancer group, and the peaks at 451 cm-1 (tryptophan) and 1441 cm-1 (lipids) were higher for the Healthy group. The PLS-DA classified the serum spectra in Healthy and Cancer groups with high accuracy (78%).

Corneal transplantation and concomitant SARS-CoV-2 infection: a case report / Transplante de córnea e infecção concomitante por SARS-CoV-2: relato de caso

ABSTRACT This report was aimed at presenting a case of neurotrophic keratitis and concomitant SARS-CoV-2 infection in a patient who has recently undergone a corneal DALK transplant. One month after corneal transplantation with adequate corneal epithelialization, the patient presented neurotrophic keratitis with a torpid course of the corneal transplant coinciding with a SARS-CoV-2 infection, with an excessive host immune response. In addition, the patient presented a re-positivization of nasopharyngeal polymerase chain reaction of SARS-CoV-2 with past disease after starting treatment with autologous serum eye drops. The implications at the ophthalmological level of SARS-CoV-2 infection may be clarified as the time the illness progresses and we learn more about how it acts. In this case, the disparity of signs and symptoms, the antecedent of corneal surgery, and the possibility of a herpetic infection as a cause of the primary leukoma suggested neurotrophic keratitis. Nonetheless, the involvement of systemic SARS-CoV-2 infection in the process, triggering an excessive host immune response at the corneal level with an increase in inflammatory cytokines must be taken into account. No relationship was found between treatment with autologous serum and re-positivization of nasopharyngeal polymerase chain reaction, presenting the patient a favorable response to treatment.

RESUMO O objetivo deste relato foi apresentar um caso de ceratite neurotrófica e infecção concomitante por SARS-CoV-2 em paciente submetido recentemente a transplante de córnea DALK. Um mês após o transplante de córnea com adequada epitelização da córnea, o paciente apresentou ceratite neurotrófica com curso tórpido do transplante de córnea, coincidindo com infecção por SARS-CoV-2, com resposta imune excessiva do hospedeiro. Além disso, o paciente apresentou repositivização da reação em cadeia da polimerase nasofaríngeo de SARS-CoV-2, com doença pregressa após iniciar tratamento com colírio de soro autólogo. As implicações a nível oftalmológico da infecção por SARS-CoV-2, podem ser esclarecidas à medida que a doença progride e aprendemos mais sobre sua forma de atuação. Neste caso, a disparidade de sinais e sintomas, o antecedente de cirurgia de córnea e a possibilidade de infecção herpética como causa do leucoma primário sugeriram ceratite neurotrófica. No entanto, deve-se levar em consideração o envolvimento da infecção sistêmica por SARS-CoV-2 no processo, desencadeando uma resposta imune excessiva do hospedeiro no nível da córnea, com aumento de citocinas inflamatórias. Não foi encontrada relação entre o tratamento com soro autólogo e a repositivização da reação em cadeia da polimerase nasofaríngea, apresentando ao paciente uma resposta favorável ao tratamento.

Insights into the Serum Metabolic Adaptations in Response to Inspiratory Muscle Training: A Metabolomic Approach Based on 1H NMR and UHPLC-HRMS/MS.

Inspiratory muscle training (IMT) is known to promote physiological benefits and improve physical performance in endurance sports activities. However, the metabolic adaptations promoted by different IMT prescribing strategies remain unclear. In this work, a longitudinal, randomized, double-blind, sham-controlled, parallel trial was performed to investigate the effects of 11 weeks (3 days·week-1) of IMT at different exercise intensities on the serum metabolomics profile and its main regulated metabolic pathways. Twenty-eight healthy male recreational cyclists (30.4 ± 6.5 years) were randomized into three groups: sham (6 cm·H2O of inspiratory pressure, n = 7), moderate-intensity (MI group, 60% maximal inspiratory pressure (MIP), n = 11) and high-intensity (HI group, 85-90% MIP, n = 10). Blood serum samples were collected before and after 11 weeks of IMT and analyzed by 1H NMR and UHPLC-HRMS/MS. Data were analyzed using linear mixed models and metabolite set enrichment analysis. The 1H NMR and UHPLC-HRMS/MS techniques resulted in 46 and 200 compounds, respectively. These results showed that ketone body metabolism, fatty acid biosynthesis, and aminoacyl-tRNA biosynthesis were upregulated after IMT, while alpha linolenic acid and linoleic acid metabolism as well as biosynthesis of unsaturated fatty acids were downregulated. The MI group presented higher MIP, Tryptophan, and Valine levels but decreased 2-Hydroxybutyrate levels when compared to the other two studied groups. These results suggest an increase in the oxidative metabolic processes after IMT at different intensities with additional evidence for the upregulation of essential amino acid metabolism in the MI group accompanied by greater improvement in respiratory muscle strength.

In-house standards derived from doping peptides: Enzymatic and serum stability and degradation profile of GHRP and GHRH-related peptides.

Matrix effect and sample pretreatment significantly affect the percentage recovery of peptides in biological matrices, affecting the method robustness and accuracy. To counteract this effect, an internal standard (IS) is used; however, in most cases this is not available, which limits the analytical method. It is important to identify short peptides that can be used as ISs in the quantification of peptides in biological matrices. In this study, doping peptides GHRP-4, GHRP-5, GHRP-6, Sermorelin (1-11), Sermorelin (13-20) and Sermorelin (22-29) were synthesized using solid-phase peptide synthesis. Treatment with human blood, trypsin and chymotrypsin was used to determine the stability of the peptides. Products were evaluated using the high-performance liquid chromatography-diode array detector (HPLC-DAD) method. The analytical methodology and sample pretreatment were effective for the analysis of these molecules. A unique profile related to protein binding and enzymatic stability of each peptide was established. GHRP-4, GHRP-6 and Sermorelin (22-29) can be considered as in-house ISs as they were stable to enzyme and blood treatment and can be used for the quantification of peptides in biological samples. Peptides GHRP-6 and Sermorelin (22-29) were used to analyse a dimeric peptide (26 [F] LfcinB (20-30)2 ) in four different matrices to test these peptides as in-house IS.

Canine visceral leishmaniasis diagnosis by UV spectroscopy of blood serum and machine learning algorithms.

Visceral leishmaniasis (VL) is a zoonotic disease caused by the protozoan Leishmania infantum, and dogs are considered the main urban hosts for future disease transmission. The first and most effective control against the spread of disease relies on identifying infected animals, followed by their treatment or sacrifice, to reduce the protozoan reservoirs. Despite the availability of various diagnostic tests for VL in dogs the development of a quick and accurate diagnosis is essential from a public health and ethical point of view. Here we analyze the use of UV-Vis spectroscopy as an alternative diagnostic method for VL diagnosis by using the antigen-antibody interaction in canine blood serum and machine learning algorithms. The main UV spectra in the 220 to 280 nm range exhibit nine electronic absorption bands, but no significative difference could be identified between the positive and negative group spectra. Finally, UV pre-proceed spectra by SNV (standard normal variate) were submitted to principal component analysis followed by Linear SVM algorithm, the prediction model was tested in a leave-one-out cross-validation and external validation test reaching an overall accuracy of 75%.

Determination of creatinine in urine and blood serum human samples by CZE-UV using on-column internal standard injection.

Creatinine is a well-stablished biomarker for kidney malfunctions and for normalization parameter of urinary quantitative information. Recently, metabolic studies have been discovering other functionalities for creatinine tests in human urine and blood serum. In this work we present an enhanced capillary electrophoresis (CE) based protocol for determination of creatinine. CE is a high-throughput separation technique that have been getting attention through the last decades and might be considered to be adopted as an analytical instrumentation for clinical purposes. In the proposed method, we performed a short injection program with on-column addition of internal standard. Additionally, the method allows a simultaneous screening of non-proteinogenic amino acids that could be considered for metabolomics purposes. We design a pilot study that successfully estimated the creatinine value in 100 urine samples with (2.85 ± 1.78) mg dL-1 LOD; (8.24 ± 5.93) mg dL-1 LOQ and 82.4% accuracy. Considering that serum creatinine is also included in the clinical laboratory routines for estimated Glomerular Filtration Rate dosage, the method was complementary applied to 10 blood serum samples, which resulted in a model with (0.4 ± 0.2) mg dL-1 LOD; (2.0 ± 0.6) mg dL-1 LOQ and 83.8% of accuracy. All results were in agreement with reference values. The proposed method promotes a great analytical frequency and reproducibility with enhanced specificity compared with the ongoing protocol by Jaffe's reaction, thereby proving to be useful as an alternative for creatinine exams that might help complete a diagnosis of a series of health-related issues.

Uso de soro autólogo no tratamento de úlcera ocular em cães: uma revisão bibliográfica / Use of autologus serum in the treatment of ocular ulcers in dogs: a bibliographic review / Uso de suero autólogo em el tratamiento de úlceras oculares em perro: una revisión bibliográfica

Sendo considerada uma doença de alto acometimento na oftalmologia veterinária de cães, a úlcera de córnea é um distúrbio caracterizado por uma ferida exposta na região corneal do olho. Os sinais são variados conforme o grau de agressividade da lesão, acarretando em perda da visão caso a intervenção não seja iniciada imediatamente. Devido a estes fatores e ao elevado nível de importância da doença, torna-se necessário o uso de terapêuticas que auxiliem o processo de reparo ocular, como a utilização do soro autólogo. Essa substância realiza uma função de evitar degradação da córnea e diminuir os sinais inflamatórios, o que torna essa prática um apoio eficaz para a finalização do tratamento, além de ser barato e de fácil acesso. Levando essas informações em consideração, o presente artigo realiza uma revisão com base em publicações apresentando o soro autólogo como um elemento adicional para o paciente poder concluir o tratamento de úlcera de córnea em menor tempo e com maior qualidade, retomando-o ao seu estado saudável.(AU)

Being considered a disease of high involvement in the veterinary ophthalmology of dogs, the corneal ulcer is a disorder characterized by an exposed wound in the corneal region of the eye. The signs vary according to the degree of aggressiveness of the lesion, leading to loss of vision if intervention is not started immediately. Due to these factors and the high level of importance of the disease, it becomes necessary to use therapies that help in the process of ocular repair, such as the use of autologous serum. This substance performs a function of preventing corneal degradation and reducing inflammatory signs, which makes this practice an effective support for the completion of treatment, besides being cheap and easily accessible. Taking this information into consideration, the present article performs a review based on publications presenting the autologous serum as an additional element for the patient to finish the corneal ulcer treatment in less time and with higher quality, returning him to his healthy state.(AU)

Siendo considerada una enfermedad de alto ataque en la oftalmología veterinaria de perros, la úlcera de córnea es una alteración caracterizada por una herida expuesta en la región corneal del ojo. Los signos son variados según el grado de agresividad de la lesión, acarreando en pérdida de la visión en caso de que la intervención no sea iniciada inmediatamente. Debido a estos factores y al alto nivel de importancia de la enfermedad, es necesario utilizar terapias que ayuden al proceso de reparación ocular, como el uso de suero autólogo. Esta sustancia cumple una función de evitar la degradación corneal y disminuir los signos inflamatorios, lo que hace de esta práctica un apoyo eficaz para el final del tratamiento, además de ser barata y de fácil acceso. Teniendo en cuenta esta información, el presente artículo realiza una revisión basada en publicaciones que presentan el suero autólogo como un elemento adicional para que el paciente finalice el tratamiento de la úlcera corneal en menor tiempo y con mayor calidad, devolviéndolo a su estado saludable.(AU)

Discrimination of leukemias and non-leukemic cancers in blood serum samples of children and adolescents using a Raman spectral model.

This study aimed to identify the differences presented in the Raman spectrum of blood serum from normal subjects compared to leukemic and non-leukemic subjects and the differences between the leukemics and non-leukemics, correlating the spectral differences with the biomolecules. Serum samples from children and adolescents were subjected to Raman spectroscopy (830 nm, laser power 350 mW; n = 566 spectra, being 72 controls, 269 leukemics, and 225 non-leukemics). Exploratory analysis based on principal component analysis (PCA) of the serum sample's spectra was performed. Classification models based on partial least squares discriminant analysis (PLS-DA) were developed to classify the spectra into normal, leukemic, and non-leukemic, as well as to discriminate spectra of leukemic from non-leukemic. The exploratory analysis showed principal components with peaks related to amino acids, proteins, lipids, and carotenoids. The spectral differences between normal, leukemic, and non-leukemic showed features assigned to proteins (serum features), amino acids, and carotenoids. The PLS-DA model classified the spectra of the normal group versus leukemic and non-leukemic groups with accuracy of 66%, sensitivity of 99%, and specificity of 57%. The PLS-DA discriminated the spectra of the leukemic and non-leukemic groups with accuracy of 67%, sensitivity of 72%, and specificity of 60%. The study showed that Raman spectroscopy is a technique that may be used for the biochemical differentiation of leukemias and other types of cancer in serum samples of children and adolescents. Nevertheless, building an extensive data library of Raman spectra from serum samples of controls, leukemics, and non-leukemics of different age groups is necessary to understand the findings better.

The influence of season, environment, sex, and body mass on biochemical profile of the freshwater turtle Phrynops geoffroanus (Schweigger, 1812) / [Influência da estação, do ambiente, do sexo e da massa corporal no perfil bioquímico do cágado Phrynops geoffroanus (Schweigger, 1812)]

We analyzed the influence of the season, the environment, and the sex, as well as the relation of body mass (BM) in the serum albumin (ALB), aspartate aminotransferase (AST), creatinine (C), creatine kinase (CK), phosphorus (P), total calcium (tCa), total protein (TP), urea (U), uric acid (UA), calcium:phosphorus ratio (Ca:P), and the globulin value (GV) of thirty individuals of Phrynops geoffroanus of the urban area of Cuiabá, Mato Grosso, Brazil. The modeling of biochemical parameters was performed using the Generalized Additive Models for Location, Scale and Shape (GAMLSS) to verify the influence of variables considered in this study on each of the biochemical parameters analyzed. The season influenced AST, CK, C, tCa, Ca:P and UA. The environment influenced tCa, Ca:P, U and UA. On the other hand, CK, tCa, P, Ca:P and U differed significantly between males and females. Regarding the BM, a relationship of this variable was observed with CK, C, tCa, P, U, UA and Ca:P. We concluded that the season, environment, sex, and body mass can influence the biochemical parameters of P. geoffroanus, and these factors should be routinely considered in the interpretation of laboratory results.

Analisou-se a influência da estação, do ambiente e do sexo, assim como a relação da massa corporal (BM) nos níveis sorológicos de albumina (ALB), aspartato aminotransferase (AST), creatinina (C), creatina quinase (CK), fósforo (P), cálcio total (tCa), sólidos totais (TS), ureia (U), ácido úrico (UA), relação cálcio:fósforo (Ca:P), e do valor da globulina (GV) de Phrynops geoffroanus da área urbana de Cuiabá, Mato Grosso, Brasil. A modelagem dos parâmetros bioquímicos foi realizada utilizando-se os modelos aditivos generalizados para locação, escala e forma (GAMLSS) para verificar a influência das variáveis consideradas neste estudo em cada um dos parâmetros bioquímicos analisados. A estação sazonal influenciou os níveis de AST, CK, C, tCa, Ca:P e UA. O ambiente foi capaz de influenciar tCa, Ca:P, U e UA. Por outro lado, CK, tCa, P, Ca:P e U diferiram significativamente entre machos e fêmeas. Em relação à BM, observou-se relação dessa variável com CK, C, tCa, P, U, UA e Ca:P. Concluiu-se que a estação sazonal, o ambiente, o sexo e a massa corporal são capazes de influenciar os parâmetros bioquímicos de P. geoffroanus e que esses fatores devem ser rotineiramente considerados na interpretação dos resultados laboratoriais.

Antibody-Based Affinity Capture Combined with LC-MS Analysis for Identification of COVID-19 Disease Serum Biomarkers.

Blood serum or plasma proteins are potentially useful in COVID-19 research as biomarkers for risk prediction, diagnosis, stratification, and treatment monitoring. However, serum protein-based biomarker identification and validation is complicated due to the wide concentration range of these proteins, which spans more than ten orders of magnitude. Here we present a combined affinity purification-liquid chromatography mass spectrometry approach which allows identification and quantitation of the most abundant serum proteins along with the nonspecifically bound and interaction proteins. This led to the reproducible identification of more than 100 proteins that were not specifically targeted by the affinity column. Many of these have already been implicated in COVID-19 disease.

Serum Soluble Mediator Profiles and Networks During Acute Infection With Distinct DENV Serotypes.

A panoramic analysis of chemokines, pro-inflammatory/regulatory cytokines, and growth factors was performed in serum samples from patients with acute DENV infection (n=317) by a high-throughput microbeads array. Most soluble mediators analyzed were increased in DENV patients regardless of the DENV serotype. The substantial increase (≥10-fold) of CXCL10, IL-6, and IFN-γ, and decreased levels of PDGF (<0.4-fold) was universally identified in all DENV serotypes. Of note, increased levels of CXCL8, CCL4, and IL-12 (≥3-9-fold) were selectively observed in DENV2 as compared to DENV1 and DENV4. Heatmap and biomarker signatures further illustrated the massive release of soluble mediators observed in DENV patients, confirming the marked increase of several soluble mediators in DENV2. Integrative correlation matrices and networks showed that DENV infection exhibited higher connectivity among soluble mediators. Of note, DENV2 displayed a more complex network, with higher connectivity involving a higher number of soluble mediators. The timeline kinetics (Day 0-1, D2, D3, D4-6) analysis additionally demonstrated differences among DENV serotypes. While DENV1 triggers a progressive increase of soluble mediators towards D3 and with a decline at D4-6, DENV2 and DENV4 develop with a progressive increase towards D4-6 with an early plateau observed in DENV4. Overall, our results provided a comprehensive overview of the immune response elicited by DENV infection, revealing that infection with distinct DENV serotypes causes distinct profiles, rhythms, and dynamic network connectivity of soluble mediators. Altogether, these findings may provide novel insights to understand the pathogenesis of acute infection with distinct DENV serotypes.

Recomendação nº 019, de 30 de junho de 2022: recomenda orientações ao Ministério da Saúde e órgãos de controle, ações sobre a situação de desabastecimento de medicamentos no SUS / Recommendation No. 019, of June 30, 2022: recommends guidelines to the Ministry of Health and control bodies, actions on the situation of drug shortages in the SUS

Recomenda ao Ministério da Saúde: I - Que realize levantamento na rede hospitalar, pública e privada, de todos os itens que se encontram em dificuldade de abastecimento; II - Que informe ao CNS, semanalmente, todos os medicamentos do Componente Especializado da Assistência Farmacêutica (CEAF) em situação de desabastecimento na rede pública, indicando os respectivos motivos, para que o Controle Social possa acompanhar a efetividade do acesso a estes medicamentos; III - Que seja provocada uma reorganização junto à indústria para estabelecer estratégias para regularização da entrega medicamentos; IV - Que o Departamento de Assistência Farmacêutica reassuma seu papel estratégico, para implementação, de forma integrada, das Políticas Nacionais de Assistência Farmacêutica e de Medicamentos; V - Que o Departamento de Assistência Farmacêutica estabeleça um plano de comunicação efetiva e transparente com o controle social; VI - Que organize a retomada dos investimentos públicos ao Complexo Econômico Industrial da Saúde como agenda estratégica para o Brasil, que perpasse pelo fortalecimento dos laboratórios oficiais, e construção de ferramentas para as Parcerias para o Desenvolvimento Produtivo (PDPs), tendo como visão estratégica a política industrial; VII - Que assuma a implementação de programas de produção de medicamentos órfãos através dos laboratórios oficiais. À Agência Nacional de Vigilância Sanitária (Anvisa) que monitore o mercado global de medicamentos e defina uma agenda regulatória de preços pela Agência. Ao Conselho Nacional de Secretários de Saúde (Conass) e ao Conselho Nacional de Secretarias Municipais de Saúde (Conasems): Que monitorem os estoques disponíveis, articulando e viabilizando os empréstimos e remanejo, quando em situação crítica em alguma localidade, bem como, realizem a articulação interfederativa para a garantia desse direito. Ao Sistema Brasileiro de Defesa da Concorrência, em especial à Superintendência-Geral do CADE: I - Que monitore o processo de editais, pregões e Atas de Registro de Preço, no intuito de garantir a prevenção e a repressão de possíveis infrações contra a ordem econômica, sejam condutas unilaterais, sejam práticas colusivas, como formação de cartéis; e II - Que considere demandar os fornecedores de medicamentos informações sobre custos de pesquisas e desenvolvimento, custos de produção e logística e políticas de desconto. Ao Programa Estadual de Proteção e Defesa do Consumidor (Procon), ao Ministério Público, à Defensoria Pública e às entidades civis de defesa do consumidor, em seus respectivos estados: Que monitorem o processo de editais, pregões, Atas de Registro de Preço e preços praticados no mercado em geral, no intuito de prevenção e repressão de possíveis infrações contra a ordem econômica em busca de prática de preços abusivos. Ao Tribunal de Contas da União (TCU) que acompanhe e monitore o processo de editais, pregões e Atas de Registro de Preço, no intuito de sanar consequências econômicas e sociais futuras, de forma que seja possível atentar ao regime de urgência da medida.

Sera from women with different metabolic and menopause states differentially regulate cell viability and Akt activation in a breast cancer in-vitro model.

Obesity is associated with an increased incidence and aggressiveness of breast cancer and is estimated to increment the development of this tumor by 50 to 86%. These associations are driven, in part, by changes in the serum molecules. Epidemiological studies have reported that Metformin reduces the incidence of obesity-associated cancer, probably by regulating the metabolic state. In this study, we evaluated in a breast cancer in-vitro model the activation of the IR-ß/Akt/p70S6K pathway by exposure to human sera with different metabolic and hormonal characteristics. Furthermore, we evaluated the effect of brief Metformin treatment on sera of obese postmenopausal women and its impact on Akt and NF-κB activation. We demonstrated that MCF-7 cells represent a robust cellular model to differentiate Akt pathway activation influenced by the stimulation with sera from obese women, resulting in increased cell viability rates compared to cells stimulated with sera from normal-weight women. In particular, stimulation with sera from postmenopausal obese women showed an increase in the phosphorylation of IR-ß and Akt proteins. These effects were reversed after exposure of MCF-7 cells to sera from postmenopausal obese women with insulin resistance with Metformin treatment. Whereas sera from women without insulin resistance affected NF-κB regulation. We further demonstrated that sera from post-Metformin obese women induced an increase in p38 phosphorylation, independent of insulin resistance. Our results suggest a possible mechanism in which obesity-mediated serum molecules could enhance the development of luminal A-breast cancer by increasing Akt activation. Further, we provided evidence that the phenomenon was reversed by Metformin treatment in a subgroup of women.

Validación de un método analítico para cuantificación de ferritina en líquido cefalorraquídeo / Validation of an analytical method for the quantification of ferritin in cerebrospinal fluid

Resumen La ferritina es una proteína de gran tamaño que se encuentra fisiológicamente en el líquido cefalorraquídeo (LCR) en concentraciones de 2-10 ng/mL. Su elevación puede utilizarse como biomarcador en distintas condiciones patológicas. El procedimiento de validación tradicional para la medición en LCR no puede ser utilizado debido a la ausencia de controles y calibradores comerciales para esta matriz. El objetivo de este trabajo fue llevar a cabo una validación analítica de ferritina en LCR. Se realizaron ensayos de estimación de precisión y veracidad mediante el protocolo EP15-A3, linealidad por el protocolo EP6-A (ambos de la guía de la CLSI), recuperación, estabilidad, contaminación por arrastre, interferencia por hemólisis y bilirrubina y límite de detección (LoD). La ferritina en LCR en el autoanalizador DxI 800 de Beckman Coulter tuvo una performance intra e interensayo <3,7%, el ensayo denota linealidad en el intervalo de 2,1-547 ng/mL; se estableció estabilidad por un período de 5 días y la recuperación resultó ser aceptable. No se evidenció efecto de contaminación por arrastre ni interferencia por hemólisis hasta un rango entre 300-500 mg/dL de hemoglobina, ni interferencia por bilirrubina hasta una concentración de 16,0 mg/dL de bilirrubina total. El LoD fue de 0,4 ng/mL. Por medio de los ensayos realizados se logró validar la ferritina en LCR a partir de la utilización de pools de muestras, lo que pudo garantizar la confiabilidad y exactitud del método analítico.

Abstract Ferritin is a large protein physiologically present in the cerebrospinal fluid (CSF) in concentrations of 2-10 ng/mL. Its elevation can be used as a biomarker in several pathological conditions. The traditional validation procedure for measurement in CSF cannot be used due to the absence of commercial controls and calibrators for this matrix. The objective of the present study was to perform CSF ferritin analytical validation. Assays such as precision and accuracy estimation through the EP15-A3 protocol, linearity according to the EP6-A protocol (both from the CLSI guidelines), recovery, stability, carry-over, hemolysis and bilirubin interference and limit of detection (LoD) were conducted. Serum samples with different concentrations of ferritin were added to aliquots of a normal CSF pool. CSF ferritin in the Beckman Coulter DxI 800 had a <3.7% intra and inter-assay performance, the assay shows linearity in the 2.1 -547 ng/mL interval, stability was established for a 5-day period and the recovery was acceptable. There was neither carry-over effect or hemolysis interference up to a range of 300-500 mg/dL of hemoglobin, nor interference by bilirubin up to 16.0 mg/dL of total bilirubin. The LoD was 0.4 ng/mL. By means of the performed assays, CSF ferritin was validated by using sample pools, thereby ensuring the reliability and accuracy of the analytical method.

Resumo A ferritina é uma grande proteína fisiologicamente encontrada no líquido cefalorraquidiano (LCR) em concentrações de 2 a 10 ng/mL. Sua elevação pode ser usada como biomarcador em diferentes condições patológicas. O procedimento de validação tradicional para medição no LCR não pode ser usado devido à ausência de controles e calibradores comerciais para esta matriz. O objetivo deste estudo foi realizar uma validação analítica da ferritina no LCR. Foram realizados estudos de precisão e veracidade utilizando o protocolo EP15-A3, linearidade pelo protocolo EP6-A (ambos das diretrizes do CLSI), recuperação, estabilidade, contaminação transportada, interferência de hemólise e bilirrubina e limite de detecção (LoD). A ferritina no LCR no DxI 800 da Beckman Coulter teve um desempenho intra e inter-ensaio <3,7%, o ensaio denota linearidade na faixa de 2,1-547 ng/mL, a estabilidade foi estabelecida em um período de 5 dias e a recuperação foi considerado aceitável. Nenhum efeito de transporte ou interferência por hemólise foi evidenciado até um intervalo entre 300-500 mg/dL de hemoglobina, nem interferência pela bilirrubina até uma concentração de 16,0 mg/dL de bilirrubina total. O LoD foi de 0,4 ng/mL. Através dos testes realizados, a ferritina no LCR foi validada, com base no uso de pool de amostras, o que poderia garantir a confiabilidade e a acurácia do método analítico.

Verificación del desempeño general del método Elecsys® Chagas en la detección de anticuerpos específicos contra Trypanosoma cruzi / Verification of the general performance of the Elecsys® Chagas method in the detection of specific antibodies against Trypanosoma cruzi

Resumen La enfermedad de Chagas es una parasitosis producida por Trypanosoma cruzi, prevalente principalmente en el continente americano, y observada en regiones no endémicas, producto de viajes y migraciones. El objetivo de este estudio fue comparar el desempeño del ensayo Elecsys® Chagas (Roche Diagnostics Alemania) (ECLIA) para el diagnóstico de la infección chagásica crónica con el método estándar y evaluar su posible empleo en reemplazo del método automatizado existente. Se estudiaron 77 muestras de sueros pertenecientes a pacientes con diagnóstico presuntivo de enfermedad de Chagas, procesadas por los distintos métodos disponibles en la Sección Parasitología del Hospital Muñiz: inmunoensayo quimioluminiscente de micropartículas (CMIA) (Abbott), enzimoinmunoanálisis de adsorción (ELISA) (Wiener) y hemaglutinación indirecta (HAI) (Lab. Lemos S.R.L.). Los resultados de los métodos ELISA y HAI fueron comparados con los obtenidos en la prueba ECLIA, y estos a su vez con el método automatizado disponible. De las muestras analizadas, 22 (28,57%) presentaron IgG anti-T. cruzi y 55 (71,43%) resultaron negativas. Con el método ECLIA se logró un 100% en los parámetros de desempeño, con diferencias en los intervalos de confianza. La razón de verosimilitud positiva y la razón de verosimilitud negativa clasificaron al ensayo como excelente y la potencia global del test apoyó esa afirmación. Los métodos inmunológicos automatizados ayudan a la performance diagnóstica en la etapa crónica de la enfermedad de Chagas, permiten minimizar errores, favorecen la velocidad de emisión de los resultados y, debido a su alta sensibilidad y especificidad, en ciertos escenarios podrían proponerse para usar como única técnica.

Abstract Chagas disease is a parasitosis caused by Trypanosoma cruzi, prevalent mainly in the American continent, and observed in non-endemic regions as a result of travel and migration. The objective of this study was to compare the performance of the Elecsys® Chagas (Roche Diagnostics Alemania) (ECLIA) assay for the diagnosis of chronic Chagas infection with the diagnostic standard, and to evaluate its possible use as a replacement for the existing automated method. A total of 77 serum samples belonging to patients with a presumptive diagnosis of Chagas disease were evaluated, processed by the different methods available in the Parasitology Section of Hospital Muñiz: microparticle chemiluminescent immunoassay (CMIA) (Abbott), enzyme-linked immunosorbent assay (ELISA) (Wiener) and indirect hemagglutination (HAI) (Lab. Lemos S.R.L). The results of the ELISA and HAI methods were compared with those obtained in the ECLIA test, and these in turn with the available automated method. Of the samples analysed, 22 (28.57%) presented IgG anti-T. cruzi and 55 (71.43%) were negative. With the ECLIA method, 100% was achieved in the performance parameters, with differences in the confidence intervals. The positive likelihood ratio and the negative likelihood ratio classify the essay as excellent, and the overall power of the test supports this statement. Automated immunological methods help diagnostic performance in the chronic stage of Chagas disease, allow minimising errors, favour the speed of issuance of results, and due to the high sensitivity and specificity, in certain scenarios, they could be proposed for use as single technique.

Resumo A doença de Chagas é uma parasitose causada pelo Trypanosoma cruzi, prevalente principalmente no continente americano, e observada em regiões não endêmicas em decorrência de viagens e migrações. O objetivo deste estudo foi comparar o desempenho do ensaio Elecsys® Chagas (Roche Diagnostics Alemanha) (ECLIA) para o diagnóstico da infecção crônica de Chagas com o método padrão e avaliar seu possível uso em substituição do método automatizado existente. Foram avaliadas 77 amostras de soro pertencentes a pacientes com diagnóstico presuntivo de doença de Chagas, processadas pelos diferentes métodos disponíveis na Seção de Parasitologia do Hospital Muñiz: imunoensaio quimioluminescente de micropartículas (CMIA) (Abbott), ensaio imunoenzimático de adsorção (ELISA) (Wiener) e hemaglutinação indireta (HAI) (Lab. Lemos S.R.L). Os resultados dos métodos ELISA e HAI foram comparados com os obtidos no teste ECLIA, e estes por sua vez com o método automatizado disponível. Das amostras analisadas, 22 (28,57%) apresentaram IgG anti-T. cruzi e 55 (71,43%) foram negativos. Com o método ECLIA, foram obtidos 100% nos parâmetros de desempenho, com diferenças nos intervalos de confiança. A razão de verossimilhança positiva e a razão de verossimilhança negativa classificam o ensaio como excelente, e a potencia geral do teste conformou essa afirmação. Os métodos imunológicos automatizados auxiliam no desempenho diagnóstico na fase crônica da doença de Chagas, permitem minimizar erros, favorecem a rapidez na emissão dos resultados e, devido à alta sensibilidade e especificidade, em determinados cenários, poderiam ser propostos para uso como técnica única.

Medición de los niveles de lactato sérico y frecuencia cardiaca en caninos (Canis lupus familiaris) braquicefálicos, mesocefálicos y dolicocefálicos sometidos a prueba de esfuerzo en trotadora motorizada / Measurement of serum lactate levels and heart rate in canines (Canis lupus familiaris) Brachycephalic, Mesocephalic and Dolichocephalic submitted to a stress test on a motorized treadmill

RESUMEN Se analizaron los niveles de lactato sérico y de frecuencia cardiaca (FC) en 15 perros clinicamente sanos que fueron sometidos a pruebas de esfuerzo en trotadora motorizada, buscando verificar un grado de fatiga muscular frente a una prueba de esfuerzo intenso. Se consideraron pacientes con morfologia craneana de tipo braquicefálico, mesocefálico y dolicocefálico. Las mediciones séricas de lactato fueron conseguidas por medio de un medidor portátil, mientras que las frecuencias cardiacas se obtuvieron a través de un oximetro portátil a los tiempos 0, 10, 20 y 30 minutos, en los diferentes momentos de esfuerzo fisico en la trotadora motorizada, la cual fue adaptada en su inclinación en 15% y a un máximo de velocidad de 10 km/h. Las medias de los niveles séricos entre perros mesocefálicos y dolicocefálicos fueron significativas, a diferencia de los perros braquicefálicos los cuales resultaron no significativos. Las medias de las frecuencias cardiacas no tuvieron diferencia significativa bajo un 95% de confianza.

ABSTRACT Serum lactate and heart rate (HR) levels were analyzed in 15 clinically healthy dogs that were subjected to stress tests on a motorized treadmill, seeking to verify a degree of muscle fatigue compared to an intense exercise test. Patients with brachycephalic, mesocephalic and dolichocephalic type cranial morphology were considered. Serum lactate measurements were procured by means of a portable meter, while heart rates were obtained through a portable oximeter at the times 0, 10, 20, and 30 minutes at different moments of physical effort on the motorized treadmill, which was adapted in its inclination by 15% and at a maximum speed of 10 km/h. Mean serum levels between mesocephalic and dolichocephalic dogs were significant, unlike brachycephalic dogs, which were not significant. The mean heart rates did not have a significant difference under 95% confidence.

Niveles séricos de la isoenzima creatina quinasa-MB y lactato deshidrogenasa como indicadores de daño miocárdico en perros con enfermedad valvular degenerativa / Serum levels of the isoenzyme creatine kinase-MB and lactate dehydrogenase as indicators of myocardial damage in dogs with degenerative valve disease

RESUMEN Se analizaron los niveles séricos de creatina quinasa-MB (CK-MB) y lactato deshidrogenasa (LDH) en 10 perros diagnosticados con enfermedad valvular degenerativa y en seis perros clinicamente sanos, con el objetivo de evaluar si sus niveles séricos indican daño miocárdico. Las muestras de suero se analizaron mediante el método UV. Se utilizó la prueba de diferenciación de medias para determinar diferencias entre medias, y la prueba de correlación de Pearson para determinar si existe correlación entre los niveles séricos de ambas enzimas. Los valores de CK-MB y de LDH fueron significativamente diferentes entre los dos grupos de pacientes. Los niveles de CK-MB y LDH tuvieron correlación positiva, pero no significativa.

ABSTRACT Serum levels of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) were analyzed in 10 dogs diagnosed with degenerative valvular disease and in six clinically healthy dogs with the objective of evaluating whether their serum levels indicate myocardial damage. Serum samples were analyzed by UV method. The mean differentiation test was used to determine differences between means and the Pearson correlation test was performed to determine if there was a correlation between the serum levels of both enzymes. The CK-MB and LDH values were significantly different between the two groups of patients. The levels of CK-MB and LDH had a positive but not significant correlation.

Diagnosing COVID-19 in human serum using Raman spectroscopy.

This study proposed the diagnosis of COVID-19 by means of Raman spectroscopy. Samples of blood serum from 10 patients positive and 10 patients negative for COVID-19 by RT-PCR RNA and ELISA tests were analyzed. Raman spectra were obtained with a dispersive Raman spectrometer (830 nm, 350 mW) in triplicate, being submitted to exploratory analysis with principal component analysis (PCA) to identify the spectral differences and discriminant analysis with PCA (PCA-DA) and partial least squares (PLS-DA) for classification of the blood serum spectra into Control and COVID-19. The spectra of both groups positive and negative for COVID-19 showed peaks referred to the basal constitution of the serum (mainly albumin). The difference spectra showed decrease in the peaks referred to proteins and amino acids for the group positive. PCA variables showed more detailed spectral differences related to the biochemical alterations due to the COVID-19 such as increase in lipids, nitrogen compounds (urea and amines/amides) and nucleic acids, and decrease of proteins and amino acids (tryptophan) in the COVID-19 group. The discriminant analysis applied to the principal component loadings (PC2, PC4, PC5, and PC6) could classify spectra with 87% sensitivity and 100% specificity compared to 95% sensitivity and 100% specificity indicated in the RT-PCR kit leaflet, demonstrating the possibilities of a rapid, label-free, and costless technique for diagnosing COVID-19 infection.

Abscesso estromal em córnea de cão / Corneal stromal abscess in a dog

Background: The eye is an important sense organ responsible for sight. The cornea is the outermost of many eye layers,being predisposed to several lesions, some of them being frequent in small animal clinics meanwhile others are unusualand seldom reported in specialized literature. The stroma abscess is a common condition in horses, but atypical in dogs,it’s a condition in which a microperforation in the corneal epithelium allows microorganisms or a foreign body to penetratethe corneal stroma, and those become inside the cornea after a re-epithelialization, producing an intense inflammatoryresponse. Therefore, this case reports a case of deep stromal abscess in a Shih Tzu dog.Case: A 4-year-old female Shih Tzu, weighing 4.6 kg, was submitted to ophthalmic examination in which the left eyepresented an opaque yellowish-white protruding structure located on the right side, in the center of the cornea (3 o´clockposition), bulbar and palpebral conjunctival hyperemia and congestion, chemosis, blepharospasm, and a moderate amountof mucous discharge. The clinical diagnosis was stromal abscess with reflex uveitis. A differential diagnosis should includeconditions affecting the anterior part of the cornea, such as an epithelial inclusion cyst and a corneal ulcer. The inclusioncyst, contrarily to the corneal abscess, doesn’t cause pain, meanwhile, the corneal ulcer was ruled out by the FluoresceinStaining Test. The recommended treatment was surgical, through lamellar keratectomy, followed by overlapping pedicledconjunctival graft and third eyelid flap. In association with the surgical treatment, oral antibiotics, anti-inflammatory,and opioids, plus topical antibiotics and autologous serum eye drop. Seventy days post-surgery, the previously observedinflammatory signs were no longer present. In the conjunctival graft insertion site perpetuated a scar leucoma with few...