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.

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)

Autologous serum eye drops diluted with 0.5% methylcellulose and 0.9% saline solution: a clinical comparative study.

PURPOSE: This clinical study compared autologous serum eye drops diluted with 0.5% methylcellulose and 0.9% saline solution. The subjective criteria for symptom improvement and the objective clinical criteria for response to therapy were evaluated. METHODS: This longitudinal prospective study enrolled 23 patients (42 eyes) with persistent epithelial defects or severe dry eye disease refractory to conventional therapy who had been using autologous serum 20% prepared with methylcellulose for > 6 months and started on autologous serum diluted in 0.9% saline solution. The control and intervention groups consisted of the same patients under alternate treatments. The subjective criteria for symptom relief were evaluated using the Salisbury Eye Evaluation Questionnaire. The objective clinical criteria were evaluated through a slit-lamp examination of the ocular surface, tear breakup time, corneal fluorescein staining, Schirmer's test, rose Bengal test, and tear meniscus height. These criteria were evaluated before the diluent was changed and after 30, 90, and 180 days. RESULTS: In total, 42 eyes were analyzed before and after 6 months using autologous serum diluted with 0.9% saline. No significant differences were found in the subjective criteria, tear breakup time, tear meniscus, corneal fluorescein staining, or rose Bengal test. Schirmer's test scores significantly worsened at 30 and 90 days (p=0.008). No complications or adverse effects were observed. CONCLUSIONS: This study reinforces the use of autologous serum 20% as a successful treatment for severe dry eye disease resistant to conventional therapy. Autologous serum in 0.9% saline was not inferior to the methylcellulose formulation and is much more cost-effective.

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.

X-ray Characterization of Conformational Changes of Human Apo- and Holo-Transferrin.

Human serum transferrin (Tf) is a bilobed glycoprotein whose function is to transport iron through receptor-mediated endocytosis. The mechanism for iron release is pH-dependent and involves conformational changes in the protein, thus making it an attractive system for possible biomedical applications. In this contribution, two powerful X-ray techniques, namely Macromolecular X-ray Crystallography (MX) and Small Angle X-ray Scattering (SAXS), were used to study the conformational changes of iron-free (apo) and iron-loaded (holo) transferrin in crystal and solution states, respectively, at three different pH values of physiological relevance. A crystallographic model of glycosylated apo-Tf was obtained at 3.0 Å resolution, which did not resolve further despite many efforts to improve crystal quality. In the solution, apo-Tf remained mostly globular in all the pH conditions tested; however, the co-existence of closed, partially open, and open conformations was observed for holo-Tf, which showed a more elongated and flexible shape overall.

Proteomic analysis of serum samples of paracoccidioidomycosis patients with severe pulmonary sequel.

BACKGROUND: Pulmonary sequelae (PS) in patients with chronic paracoccidioidomycosis (PCM) typically include pulmonary fibrosis and emphysema. Knowledge of the molecular pathways involved in PS of PCM is required for treatment and biomarker identification. METHODOLOGY/PRINCIPAL FINDINGS: This non-concurrent cohort study included 29 patients with pulmonary PCM that were followed before and after treatment. From this group, 17 patients evolved to mild/ moderate PS and 12 evolved severe PS. Sera from patients were evaluated before treatment and at clinical cure, serological cure, and apparent cure. A nanoACQUITY UPLC-Xevo QT MS system and PLGS software were used to identify serum differentially expressed proteins, data are available via ProteomeXchange with identifier PXD026906. Serum differentially expressed proteins were then categorized using Cytoscape software and the Reactome pathway database. Seventy-two differentially expressed serum proteins were identified in patients with severe PS compared with patients with mild/moderate PS. Most proteins altered in severe PS were involved in wound healing, inflammatory response, and oxygen transport pathways. Before treatment and at clinical cure, signaling proteins participating in wound healing, complement cascade, cholesterol transport and retinoid metabolism pathways were downregulated in patients with severe PS, whereas signaling proteins in gluconeogenesis and gas exchange pathways were upregulated. At serological cure, the pattern of protein expression reversed. At apparent cure pathways related with tissue repair (fibrosis) became downregulated, and pathway related oxygen transport became upregulated. Additionally, we identified 15 proteins as candidate biomarkers for severe PS. CONCLUSIONS/SIGNIFICANCE: Development of severe PS is related to increased expression of proteins involved in glycolytic pathway and oxygen exchange), indicative of the greater cellular activity and replication associated with early dysregulation of wound healing and aberrant tissue repair. Our findings provide new targets to study mechanisms of PS in PCM, as well as potential biomarkers.

Serum Analysis of Women with Early-Stage Breast Cancer Using a Mini-Array of Tumor-Associated Antigens.

Background: Several studies have shown that patients with cancer have antibodies in serum that react with cellular autoantigens, known as Tumor-Associated Antigens (TAA). The present work aimed to determine whether a mini-array comprising four recombinant TAA increases the detection of specific serum antibodies for the diagnosis of early-stage breast cancer. Methods: The mini-array included Alpha 1-AntiTrypsin (A1AT), TriosePhosphate Isomerase 1 (TPI1), Peptidyl-Prolyl cis-trans Isomerase A (PPIA), and PeroxiReDoXin 2 (PRDX2) full-length recombinant proteins. The proteins were produced after gene cloning, expression, and purification, and were verified by Western blot assays. Then, Dot-Blot was performed to find antibodies against the four TAA in 12 sera from women with early-stage breast cancer (stage II) and 12 sera from healthy women. Results: Antibody detection against individual TAA in early-stage breast cancer sera ranged from 58.3% to 83.3%. However, evaluation of the four TAA showed that there was a positive antibody reaction reaching a sensitivity of 100% and a specificity of 85% in early-stage breast cancer, suggesting that this mini-array must be evaluated as a clinical diagnostic tool for early-stage breast cancer in a larger sample size. Conclusion: Our results suggest that TAA mini-arrays may provide a promising and powerful method for improving the detection of breast cancer in Mexican women.

Serum biochemistry of the Arrau turtle (Podocnemis expansa): Frozen analyte stability and the effects of long-term storage.

BACKGROUND: Turtles are a major source of protein for riverside human populations in Brazil. The encouragement of commercial breeding meets conservation efforts for these animals, and it is, therefore, crucial to understand the physiologic and behavioral aspects of semi-aquatic species in captive conditions. Serum biochemical tests are ancillary diagnostic tools, and sample storage is a main problem since clinical laboratories are not always available near the habitats of these species. OBJECTIVES: The aim of this study was to provide information about the stability of albumin, aspartate aminotransferase (AST), calcium, creatinine kinase (CK), total cholesterol (Chol), alkaline phosphatase (ALP), gamma glutamyltransferase (GGT), total protein (TP), and urea at different storage times. METHODS: In all, 17 Arrau turtles (Podocnemis expansa) were used, and the serum obtained was separated into aliquots and analyzed at 0, 4, 8, 16, and 32 days after being stored at -20°C. RESULTS: The results showed that albumin, AST, CK, GGT, and TP suffered interference due to the long storage times. CONCLUSION: Analytes such as ALP, calcium, Chol, and urea can be evaluated for up to 1 month after freezing. Albumin, AST, and TP can be analyzed up to 1 week after freezing without alterations, and CK GGT are best evaluated on fresh samples.

Simultaneous extraction of antibiotic and estrogen from animal blood serum using aqueous two-phase systems as predictor of environmental impact.

Efficient analytical methods are required for optimizing dosage of veterinary antibiotics and hormones in order to reduce toxicity and antimicrobial resistance in the environment. Thus, the objective of this work was to develop a rapid and low-cost method for determination of hormone estradiol and antibiotic chlortetracycline in bovine and porcine blood serum by aqueous two-phase system (ATPS) extraction and capillary electrophoresis quantification. ATPS based on ionic liquid cholinium alaninate and citrate salt along with mixtures of protic and aprotic polar solvents were evaluated in terms of recovery of extraction (%R). The liquid-liquid equilibrium, phase diagrams, and tie lines are discussed. Antibiotic migrated to solvent-rich phase (R ≈ 89.0%) to all systems. Estradiol migrates to ionic liquid-rich phase; however, addition of 10% methanol changed partition to solvent-rich phase (R ≈ 89.7%). The method has high recovery and cleanliness, is cost-efficient, scalable, and hence is adequate for screening of antibiotics and hormones tested in animal blood serum for dosage optimization and to predict their environment.

Assessment of renal functions and lesions in dogs with serological diagnosis of canine visceral leishmaniasis

Background: Visceral leishmaniasis is a complex vector-borne disease caused by the protozoan Leishmania infantum. In urban centers of South America, where this zoonotic cycle occurs, dogs seem to be the main reservoirs and infection sources. Animals with canine visceral leishmaniasis (CVL) may have a wide clinical spectrum, and dogs are usually classified as asymptomatic, oligosymptomatic, and symptomatic. Several organs are affected in canine CVL, and renal involvement is often a determining factor in dog prognosis. Nevertheless, serum markers are slow to indicate loss of renal function. The aim of this study was to evaluate kidney impairment in dogs diagnosed with CVL. Materials, Methods & Results: Blood and urine samples were collected from 45 dogs from Barra Mansa-RJ, and used for urinalysis, urine protein/creatinine (UPC) ratio, and serum concentrations of urea and creatinine. The animals were classified as symptomatic (42.2%), oligosymptomatic (37.8%), and asymptomatic (20.0%). Some alterations were found in the urine samples; pale-yellow color in 17.8%, low specific gravity in 6.7%, turbidity in 51.1%, proteinuria in 80%, occult blood in 46.7%, bilirubin in 8.89%, and glucose in 6.7% of the samples. According to the UPC ratio, 60% of dogs were proteinuric, and UPC > 2.0 was high in symptomatic dogs. Azotemia was observed only in three dogs with CVL. Discussion: The majority of dogs presented one or more symptoms of CVL, as expected in an endemic area from Brazil. Pale-yellow urine was observed in some samples, and this change, when accompanied by the decreased urine specific gravity in dogs with CVL, suggests some degree of kidney disease. The presence of epithelial and red blood cells, leukocytes, bacteria, suspended mucus, and phosphate crystals that precipitate in alkaline urines could be associated, to some degree, with the urine turbidity found in the present study. The alkaline urine identified in some dogs could be related to...

Metabolic profiling of serum in patients with cartilage tumours using 1 H-NMR spectroscopy: A pilot study.

Cartilage-forming lesions include tumours that can vary in severity from benign enchondromas to high-grade malignant chondrosarcomas. Chondrosarcoma is the second most frequent malignant bone tumour, accounting for 20-30% of all malignant bone neoplasms. Surgery is the standard treatment for cartilage tumours (CTs); however, their incidental diagnosis and the difficult differentiation of low-grade lesions like chondrosarcoma grade I from benign entities like enchondroma are challenges for clinical management. In this sense, the search for circulating biomarkers for early detection and prognosis is an ongoing interest. Targeted metabolomics is a powerful tool that can propose potential biomarkers in biological fluids as well as help to discover disturbed metabolic pathways to reveal tumour pathogenesis. In this context, the aim of this study was to investigate the 1 H nuclear magnetic resonance metabolomic serum profile of patients with CTs contrasted with healthy controls. Forty-one metabolites were identified and quantified; the multivariate statistical methods principal component analysis and partial least squares discriminant analysis reveal a clear separation of the CT group, that is, the differential metabolites that were involved in two main metabolic pathways: the taurine and hypotaurine metabolism and synthesis and degradation of ketone bodies. Our results represent preliminary work for emergent serum-based diagnostics or prognostic methods for patients with chondrogenic tumours.

Serum analysis in women and in vitro skin assay for the assessment of exposure to parabens in antiperspirants.

The present study assessed the exposure to methylparaben (MP) and propylparaben (PP) from antiperspirants in serum of 24 women aged 20-30 years old and an in vitro skin assay. An effective liquid chromatography-tandem mass spectrometry method for the determination of MP and PP levels in serum was developed and validated in the range of 10-100 µg/L; the method was fast, simple, sensitive, linear, precise, and accurate. In addition, a simple and rapid liquid chromatography-ultraviolet detection method for the determination of MP and PP levels in antiperspirants was developed and validated in the range of 2-26 mg/L, which presented satisfactory linearity, precision, and accuracy. Using these two methods, 20 commercial antiperspirants were evaluated, and only three showed MP and PP in the formulation. The antiperspirant containing 0.2% and 0.1% w/w MP and PP, respectively, was given to the volunteers, to estimate the internal dose, and submitted to a pig ear skin permeation assay in Franz diffusion cells, presenting a permeation flux of 32% for MP and 71% for PP. In this assay, both MP and PP permeated the skin; however, there was no correlation between antiperspirant use and paraben serum concentration in the volunteers. Graphical abstract.

Polyacrylamide-alginate (PAAm-Alg) and phospho-L-tyrosine-linked PAAm-Alg monolithic cryogels: Purification of IgG from human serum.

In recent decades cryogels as monolithic materials have gained interest as stationary phase in chromatography for purification of biomolecules. In this study, polyacrylamide-alginate (PAAm-Alg) monolithic cryogels were prepared by cryo-copolymerization of acrylamide and alginate monomers and methylene-bisacrylamide as crosslinker to be used as a matrix in affinity chromatography for purification of proteins. Ortho-phospho-L-tyrosine (P-Tyr) was covalently attached onto PAAm-Alg cryogels via bisoxirane-activation (PAAm-Alg-Bix-P-Tyr) and both derivatized and non-derivatized cryogels were utilized for the purification of immunoglobulin G (IgG) from human serum. Cryogels were characterized by scanning electron microscopy, swelling tests, elemental analysis, FTIR, and flow dynamics. The effects of buffer systems, conductivity, and pH on IgG adsorption were studied. Through breakthrough curve analysis a dynamic capacity of 9.2 mg IgG/mL with an IgG purity of 94% was obtained (based on ELISA analysis of IgG and albumin) for PAAm-Alg-Bix-P-Tyr cryogel when human serum was diluted in 10 mmol/L NaP buffer at pH 6.0. The adsorption isotherm data were well described by the Langmuir model with value of maximum adsorption capacity of 36.12 ±â€¯3.63 mg of IgG/g for PAAm-Alg-Bix-P-Tyr. The PAAm-Alg-Bix-P-Tyr cryogel provides an attractive alternative for adsorption of IgG from human serum.

The Effects of Supplementing Hancornia speciosa (Mangaba) on Bowel Motility and Inflammatory Profile of Wistar Rats.

The prevalence of gastrointestinal disorders, such as constipation, has been increasing. Genetic factors and lifestyle are some of the etiologies of this issue, affecting the health of the population. Natural products have properties that contribute to health maintenance and health promotion, including reduction of the inflammatory process. Hancornia speciosa, popularly known as mangaba, is an abundant and native fruit in the Brazilian Cerrado, commercialized for culinary purposes and used because of its pharmacological properties. The objective of this study was to evaluate if the supplementation of different concentrations of mangaba pulp can improve intestinal motility and bowel health in Wistar rats. Forty male rats were divided into five groups. The experiment lasted 14 days and the groups were tested with water, industrialized laxative jelly made from tamarind as medication, or mangaba at 5, 10, and 15 mL/kg of body weight. Food intake, weight gain, ion balance, intestinal motility, and histopathological analysis of the small intestine, large intestine, and liver were evaluated. Supplementation of mangaba pulp at its highest concentration (15 mL/kg body weight) caused a 15% increase in the distance traveled by the charcoal meal, and a decrease in serum magnesium levels and white cells in both the small and large intestines. The results suggest that mangaba pulp presents laxative, anti-inflammatory properties and that its consumption is beneficial and should be encouraged.

Non-Enzymatic Impedimetric Sensor Based on 3-Aminophenylboronic Acid Functionalized Screen-Printed Carbon Electrode for Highly Sensitive Glucose Detection.

A highly sensitive glucose sensor was prepared by a one-step method using 3-aminophenyl boronic acid as a unit of recognition and a screen-printed carbon electrode (SPCE) as an electrochemical transducer. Scanning Electron Microscopy confirmed the success of the functionalization of the SPCE due to the presence of clusters of boronic acid distributed on the carbon surface. In agreement with the Electrochemical Impedance Spectroscopy (EIS) tests performed before and after the functionalization, Cyclic Voltammetry results indicated that the electroactivity of the electrode decreased 37.9% owing to the presence of the poly phenylboronic acid on the electrode surface. EIS revealed that the sensor was capable to selectively detect glucose at a broad range of concentrations (limit of detection of 8.53 × 10-9 M), not recognizing fructose and sucrose. The device presented a stable impedimetric response when immediately prepared but suffered the influence of the storage time and some interfering species (dopamine, NaCl and animal serum). The response time at optimized conditions was estimated to be equal to 4.0 ± 0.6 s.

Serum concentration of acute phase proteins and cytokines in vaccinated pigs challenged with foot-and-mouth disease virus serotype O

The present study aimed to investigate the kinetics of acute phase proteins and cytokines in the serum of immunized or naive pigs following infection with a field isolate of foot-and-mouth disease (FMD) virus. Five-month-old SPF pigs were intramuscularly immunized with or without the 146S antigen (7.5 µg per head) purified from FMD virus (O/Andong/SKR/2010) and were subsequently challenged 30 days post-vaccination (dpv) with FMD virus on each footpad. Serum samples were obtained at 28 dpv and on days 1, 3, and 5 post-infection (dpi). At 28 dpv, the serum concentrations of C-reactive protein and tumor necrosis factor alpha (TNF-α) were higher in the immunized pigs compared with the non-immunized control pigs. Upon challenge, the levels of all measured acute phase proteins (i.e., C-reactive protein, major acute phase protein, S-amyloid A, and haptoglobin) plateaued at 1 dpi in the immunized pigs whereas they gradually increased up to 5 dpi in the non-immunized control pigs. Levels of serum TNF-α remained higher in the immunized challenged pigs compared with the non-immunized challenged pigs. The serum concentration of interleukin (IL)-6 in the immunized challenged pigs was undetectable at all time points, but moderately increased upon challenge in the non-immunized challenged pigs. IL-1ß was not detected in any of the pigs at any of the observed time points. Collectively, these findings show that the immunized pigs exhibit a rapid serum acute phase protein response following subsequent challenge with the field FMD virus compared with the non-immunized pigs.(AU)

Choosing sample sizes for various blood parameters of broiler chickens with normal and non-normal observations.

Experimental power is a measure of the ability of an experiment to detect differences between treatment means. Researchers design experiments and then calculate the probability that differences are simply due to chance, the null hypothesis. The objective of the analyses reported here was to determine the appropriate number of samples to demonstrate significant differences of various magnitudes from broiler chicken blood constituents. Over 800 samples were taken for a study of the effects of sample storage time, serum vs. plasma, light intensity, and fed vs. fasted birds on blood cholesterol, triglycerides, uric acid, glucose, total protein (TP), albumin, globulin, alanine aminotransferase (ALT), aspartate aminotransferase, gammaGT, creatinine, alkaline phosphatase, Ca and P. Various transformations increased the QQ plot R2 values from 0.000 to 0.149 or 0.00 to 17.62%. Most of the QQ plot R2 values were at or above 0.90. The 1/x2 transformation of blood P data showed the biggest increase in QQ plot R2 (0.846 to 0.995). The different standard deviations and coefficients of variation (CVs) found for each variable resulted in widely different numbers of replicates needed to detect differences in 2 treatment means. The extremes were glucose with a CV of 6.9% and ALT with a CV of 39.7%. For glucose, 15 replicates are needed to find a 10% difference in 97% of experiments; for ALT, 15 replicates would detect a 50% difference 91% of the time. The use of parameters such as cholesterol, glucose, TP, albumin, and globulin showed low CVs, indicating they may be considered as stable parameters. The lower CVs make it possible to find differences with a smaller number of replicates used in studies. As reported, the phosphorus values did not have a normal distribution of the data, so a transformation of these data could be an alternative to better discuss the results found.

The correlation of TNF alpha levels with the lipid profile of dengue patients.

Immunological factors, such as cytokines, have been proposed as a cause of changes in the lipid profile of dengue patients. We studied whether serum lipid levels and serum TNF-α levels are associated in a group of dengue patients from an endemic region in the Northwest of Mexico. We found statistically important differences in the serum lipid profile and the TNF-α levels of dengue patients compared with the control group, were observed. However, TNF-α levels did not correlate with the lipid profile of dengue patients.