Association of bull semen protein estimates and SDS-PAGE profiles on semen freezability

It is vital to identify the ejaculate with good freezability by determining the biochemical makeup of the ejaculate at the pre-freeze stage. The present study targeted to assess the use of the protein estimates and profiles at the pre-freeze stage as markers of freezability in Frieswal populations. Storing the proteins for proteomic studies is always tricky in the case of animal studies, where accessibility to liquid nitrogen is limited. Hence alternative storing approaches need to be optimized. The second part of this study examined the protein concentration and protein profiles of RNALater and frozen stored sperm cells to assess the use of RNALater preservation in sperm proteomic studies. Sperm and seminal plasma protein concentrations were quantified using Bradford assay, and total protein quantities were derived. The seminal plasma and sperm protein profiles were generated with SDS-PAGE. The protein estimates and SDS-PAGE profiles of good and poor freeze-groups were similar. Also, sperm and seminal plasma protein concentration were not correlated with the semen volume and sperm count. Even though the yield was comparatively less, the protein profiles of sperm preserved by RNALater were similar to that of frozen sperms. The present study results indicate that the protein estimates and qualitative profiles of sperm and seminal plasma proteins may not be sufficient to reveal the differences in the proteome of good and poor freezable bulls at the macro level. Hence, the protein estimates and profiles of neat semen may not be helpful for the prediction of freezability at the pre-freeze stage. Secondly, this study indicates that RNALater preservation helps store sperms for proteome analysis studies.

Association of genetic markers with cardiomyopathy.

Background: Cardiomyopathy is an anatomic and pathologic diagnosis associated with muscle or electrical dysfunction of the heart. Cardiomyopathies represent a heterogeneous group of diseases that often lead to progressive heart failure with significant morbidity and mortality. Cardiomyopathy and myocarditis resulted in 443,000 deaths in 2013 up from 294,000 in 1990. Objective: The main objective of the present study is to observe the association of cardiomyopathy and genetic markers such as red cell enzymes namely, Esterase D [ESD] and Super oxide dismutase [SOD] and plasma proteins namely, Haptoglobin [HP] and Group specific component [GC] systems. Methods: In the present study, fifty cases presenting cardiomyopathy and fifty cases of age and sex matched healthy controls were included. Red cell enzymes were determined by standard agarose gel electrophoresis. Plasma samples were typed using PAGE electrophoresis. The statistical significance of differences between patients and controls were tested. Analysis of the data was carried out using Epi Info 5 software. Relative risk was calculated by the random-effects method. For odds ratio, confidence interval was calculated. The significance level was 5%. Results: The inter group heterogeneity for ESD and SOD of red cell enzymes and GC system of plasma proteins was found to be a significant value (ESD: χ2 =10.2564; d.f. = 2; 0.01>p>0.001; SOD: χ2 = 11.1120; d.f. = 2; 0.01>p>0.001; GC: χ2 = 15.5044; d.f. = 2; p>0.001), when observed between cardiomyopathy patients and controls. Thus, all the examined groups were deviating from Hardy-Weinberg equilibrium indicating a significant association between cardiomyopathy and these red cell enzymes and plasma protein markers. There was a predominant occurrence of Haptoglobin 2 phenotype in patients when compared to controls. Risk estimates show significant association with both ESD and GC systems with an increased risk of 100% and more, indicating that individuals with ESD (2-2 and 2-1) and GC (2-1) phenotypes are more likely to get the disease when compared with the other phenotypes of the ESD and GC systems. Conclusions: Out of seven genetic markers, four markers (ESD, SOD, HP and GC) are found to be significant i.e. they show some relation with the cardiomyopathy which influences the disease. Furthermore studies on genetic markers, to be attempted in future, would certainly enlighten us to assess the role of these polymorphic systems in different cardiomyopathies.

Advanced Glycation End Products and Diabetic Complications

During long standing hyperglycaemic state in diabetes mellitus, glucose forms covalent adducts with the plasma proteins through a non-enzymatic process known as glycation. Protein glycation and formation of advanced glycation end products (AGEs) play an important role in the pathogenesis of diabetic complications like retinopathy, nephropathy, neuropathy, cardiomyopathy along with some other diseases such as rheumatoid arthritis, osteoporosis and aging. Glycation of proteins interferes with their normal functions by disrupting molecular conformation, altering enzymatic activity, and interfering with receptor functioning. AGEs form intra- and extracellular cross linking not only with proteins, but with some other endogenous key molecules including lipids and nucleic acids to contribute in the development of diabetic complications. Recent studies suggest that AGEs interact with plasma membrane localized receptors for AGEs (RAGE) to alter intracellular signaling, gene expression, release of pro-inflammatory molecules and free radicals. The present review discusses the glycation of plasma proteins such as albumin, fibrinogen, globulins and collagen to form different types of AGEs. Furthermore, the role of AGEs in the pathogenesis of diabetic complications including retinopathy, cataract, neuropathy, nephropathy and cardiomyopathy is also discussed.

Método FAMACHA©, escore corporal e de diarreia como indicadores de tratamento anti-helmíntico seletivo de ovelhas em reprodução / FAMACHA© system, body condition score and diarrhea score as indicators for the targeted selective anthelmintic treatment of breeding ewes

As parasitoses gastrintestinais constituem o principal entrave sanitário à ovinocaprinocultura, principalmente devido à resistência aos anti-helmínticos. Uma das formas de limitar a progressão da resistência aos fármacos é evitar a sua utilização em lotes ou rebanhos inteiros, restringindo seu uso aos animais que necessitam tratamento (tratamento seletivo). O objetivo deste trabalho foi estudar o uso dos escores corporal (EC) e de diarreia (ED) como indicadores do tratamento seletivo em ovinos, em comparação com o método FAMACHA© (GF) e indicadores laboratoriais de parasitismo. Foram acompanhadas quinzenalmente 15 ovelhas em lactação (agosto-novembro 2009) e após o desmame (dezembro 2009 ­ fevereiro de 2010) pelo GF, EC, ED, e análises laboratoriais para determinação do hematócrito (HT), proteínas plasmáticas totais (PPT) e contagem de ovos de helmintos por grama de fezes (OPG). A média de OPG se manteve estável, exceto no segundo mês de lactação (média 6057,69 OPG). Durante todo o acompanhamento, o GF foi moderadamente correlacionado ao OPG (0,37); EC (-0,32); PPT (-0,34) e HT (-0,54). Para o EC, os valores de correlação com o OPG foram de -0,35; -0,32 com GF; 0,45 com PPT e de 0,50 com HT. O ED não se mostrou um bom indicador indireto de verminose, pois não foi obtida nenhuma correlação significativa com as outras variáveis estudadas. Como o escore corporal foi correlacionado às variáveis laboratoriais em índices semelhantes aos obtidos pelo método FAMACHA© , seu uso é sugerido como ferramenta adicional de tratamento seletivo para ovelhas em lactação e nos meses subsequentes ao desmame.

The gastrointestinal parasitic infections constitute the main obstacle to the health of sheep and goat, mainly due to anthelmintic resistance. An alternative to limit the progression of drug resistance is to avoid to deworm the hole heard or group, treating individually only those animals that require so (selective treatment). The goal of this work was to study the condition score (BS) and the diarrhea score (DS) as indicators of selective treatment in sheep, in comparison to the FAMACHA© system (EF) and to laboratory analyses that indicate parasitism. Fifteen lactating (Aug.- Nov. 2009) or post weaning (Dez. 2009- Feb.2010) ewes were sampled fortnightly for EF, BS, DS and also for the laboratorial analysis to determine packed cell value (PCV/HT), total plasma protein fraction (TPP) and faecal egg counts (FEC/OPG). The mean FEC remained stable except in the second month of lactation period (average 6057.69 EPG). Throughout the monitoring, the GF was moderately correlated with FEC (0.37), BS (-0.32), TPP (-0.34) and PCV (-0.54). For the BS, the value of correlation with FEC was -0.35, -0.32 to GF; 0.45 to TPP and 0.50 to PCV. The DS was not a good indicator of parasite infection, because there was no significant correlation obtained with the other variables. As the body condition score was correlated to the laboratory variables in similar indices of the FAMACHA© system, we suggest the its use as an additional tool of selective treatment for ewes during lactation period and after weaning.

Topografia de ligação de proteínas do plasma seminal à membrana de espermatozoides bovinos epididimários e ejaculados / Binding patterns of seminal plasma plasma proteins on bovine epididymal and ejaculated sperm membrane

The present study was designed to investigate the topographical distribution of seminal plasma (SP) proteins on epididymal and ejaculated bovine sperm. Using immunocytochemistry and confocal microscopy the binding patterns of bovine SP proteins BSP-A3, albumin, transferrin, prostaglandin D-synthase (PGDS) and nucleobindin in ejaculated and cauda epididymal sperm from adult bulls were evaluated. Experiments were performed using sperm from 5 males. Data showed a positive signal, only detected for anti-PGDS, in the acrosomal cap of epididymal and ejaculated sperm. In ejaculated sperm, a very weak signal for nucleobindin 2 in the midpiece and equatorial regions was detected, using the anti-rat nucleobindin. BSP-A3 was detected on all sperm regions studied, with a more evidenced signal in acrosome and midpiece. However, no binding was detected for albumin or transferrin in neither epididymal nor ejaculated sperm. In conclusion, PGDS, BSP-A3 and nucleobindin interact directly with bovine sperm, with specific topographic distribution. These findings may add to the knowledge of how these proteins modulate sperm functions, thus providing fundamental support for studies designed to evaluate how they influence sperm functions.

Investigou-se a distribuição topográfica da ligação de proteínas seminais à membrana de espermatozoides bovinos epididimários e ejaculados. Utilizando imunocitoquímica e microscopia confocal, avaliaram-se a topografia de ligação das proteínas BSP-A3, albumina, transferrina, prostaglandina D sintetase (PGDS) e nucleobindina 2 (NUC2) à membrana espermática. Os experimentos foram realizados utilizando espermatozoides de cinco touros. Os resultados mostraram que, para espermatozoides epididimários, somente detectou-se a PGDS na crista do acrossomo. Nos espermatozoides ejaculados, a PGDS ligou-se de forma mais intensa à crista acrossômica, enquanto a NUC2 apresentou sinal bastante fraco na peça intermediária e região equatorial. A BSP-A3 ligou-se a todas as regiões estudadas, de forma mais intensa na peça intermediária e acrossomo. Nenhum sinal foi detectado para albumina ou transferrina, seja em espermatozoides epididimários ou ejaculados. Concluiu-se que PGDS, BSP-A3 e NUC2 interagem diretamente com espermatozoides bovinos, e mostrou distribuição topográfica específica. Estes achados permitem melhor compreensão sobre o papel desempenhado por essas proteínas na regulação da função espermática e da fertilidade.

Quadro seroproteico como auxílio diagnóstico na anemia hemolítica imunomediada em cães / Serum proteic profile as diagnosis aids in immune-mediated hemolytic anemia in dogs

No presente protocolo experimental, determinaram-se os proteinogramas séricos, por intermédio da eletroforese em gel de poliacrilamida contendo duodecil sulfato de sódio (SDS-PAGE), de 120 cães com raças e idades variadas e atendidos junto ao Hospital Veterinário "Governador Laudo Natel" da FCAV/Unesp, com o objetivo principal de comparar diferentes frações seroproteicas em estados anêmicos regenerativos, arregenerativos, imunomediados primários e secundários. Os referidos animais foram distribuídos em cinco grupos experimentais: grupo 1: 20 cães de controle; grupo 2: 28 cães com anemia regenerativa não imune; grupo 3: 27 cães com anemia arregenerativa não imune; grupo 4: 10 cães com anemia hemolítica imunomediada primária; grupo 5: 35 cães com anemia hemolítica imunomediada secundária. A técnica SDS-PAGE permitiu o fracionamento de 24 proteínas, cujos pesos moleculares (PM) variaram de 18.000 a 165.000 daltons (Da). Os cães com AHIM primária e secundária apresentaram 24 frações proteicas em seus traçados eletroforéticos, enquanto que cães de controle (1) e portadores de anemia regenerativa (2) e arregenerativa (3) de natureza não imune apresentaram 23 frações de proteínas, cuja proteína de peso molecular 68.000Da não foi encontrada. Dessa forma, 23 frações proteicas foram detectadas e revelaram-se comuns aos proteinogramas dos cães de controle e daqueles dos quatro grupos experimentais. Destas, identificaram-se nominalmente 11 frações proteicas, e as demais foram estudadas com base nos seus respectivos pesos moleculares. Em relação aos cães de controle, os anêmicos (grupos 2, 3, 4 e 5) apresentaram maiores concentrações de transferrina sérica e entre estes os animais portadores da AHIM primária. Todos os cães anêmicos apresentaram teores séricos de haptoglobina e fosforilase significativamente maiores que os controles, enquanto que a concentração sérica de ceruloplasmina foi significativamente maior nestes. Tais achados analisados em...

This assay aimed to determine the serum protein - via polyacrylamide gel electrophoresis, which contained duodecil sodium sulfate (SDS-PAGE) - in 120 dogs, with different breeds and ages, seen by the Veterinary Hospital "Governor Laudo Natel." These animals were grouped into five experimental groups: Group 1 - group control with 20 dogs, group 2 - 28 dogs with regenerative anemia; group 3 - 27 dogs with arregenarative; anemia group 4 - 10 dogs with primary immune-mediated hemolytic anemia (AHIM 1.rd); group 5 - 35 dogs with secondary immune-mediated hemolytic anemia (AHIM 2.rd). The technique allowed the SDS-PAGE fractionation of 24 protein, whose molecular weights (PM) ranged from 18,000 to 165,000 daltons (Da). The dogs with 1st and 2nd AHIM showed 24 protein fractions in their tracks electrophoretic, while other groups of dogs showed 23 fractions of protein, whose molecular protein weight of 68,000Da was not found. Thus, twenty-three proteins were common to proteinograms of the five experimental groups. From these, it was possible to identify eleven protein fractions nominally, and others were identified by their molecular weights. For control dogs, the anemic (groups 2, 3, 4 and 5) showed higher concentrations of serum transferrin and between them, the animals carrying the primary IMHA. All groups of dogs showed anemic levels of serum haptoglobin and phosphorylase significantly higher than the control dogs, while the serum ceruloplasmin was lower in anemic dogs. These findings provide additional information to the elucidation of the AHIMs in dogs.

Evaluation of Serum Levels of Hepatic Enzymes and Plasma Proteins between Different Types of Acute Viral Hepatitis.

This cross-sectional study was carried out in 80 serologically diagnosed cases of acute viral hepatitis to assess and compare the serum hepatic enzymes & plasma proteins between four different types (A,B,C,E), 20 in each group. Hepatitis E, hepatitis B and hepatitis C were more prevalent in males than that in females. The study showed that geometric mean of S.AST of all the four types differed significantly (F= 274.94, p<0.001). Geometric mean of S.ALT, S.AST and S.ALP in cases of HCV were significantly lower than others (p<0.001). Geometric mean of S.ALT & S.AST in cases of HEV were significantly increased than others (p<0.001). But the geometric mean of S.ALP of HBV was significantly higher than others (p<0.001). On the other hand though S.ALP of HAV and HEV was lower than HBV but significantly higher than HCV (p<0.001). The mean±SD of serum albumin of HCV was decreased significantly in contrast to those of HAV and HBV (p<0.001). A:G ratio of HCV was also significantly lower than other three (p<0.001). It was revealed through the study that hepatic enzymes were most affected in cases of HEV but least affected in cases of HCV.

Característica leucocitária, relação albumina/globulina, proteína plasmática e fibrinogênio de bovinos da raça Nelore, confinados e terminados a pasto / Leukocyte characteristic, albumin/globulin relation, plasmatic protein and fibrinogen of bovines of the Nelore race confined and grazing

Esse trabalho avaliou as mudanças na contagem de leucócitos e algumas proteínas séricas de bovinos confinados e terminados a pasto. De 120 amostras sangüíneas coletadas, 60 foram obtidas de bovinos Nelores machos confinados e 60 de animais com as mesmas características, porém manejados extensivamente. As amostras foram obtidas por ocasião do abate desses animais. Os parâmetros estudados foram contagem de leucócitos, razão albumina/globulina e concentração de fibrinogênio plasmático. Na análise dos dados empregou-se estatística descritiva, obtendo-se as médias, desvio padrão e coeficiente de variação para todos as variáveis avaliadas e posteriormente comparou-se as médias por meio de teste não-paramétrico. Os bovinos terminados a pasto apresentaram maior nível de globulina e fibrinogênio (P>0,05) quando comparados com os confinados (globulina: pastejo=3,29g dL-1 0,76; confinamento 2,99g dL-1±0,60 e Fibrinogênio: pastejo=872mg dL-1±610; confinamento=633mg dL-1±319). O número de leucócitos total foi de 7,64±2,15 em bovinos confinados e de 7,72±1,84 nos terminados a pasto. Não houve diferença (P>0,05) entre essa variável e a contagem diferencial de leucócitos bem como na proteína sérica total (g dL-1) dos bovinos terminados a pasto (6,10±0,53) e dos confinados (5,96±0,49). O nível de albumina dos bovinos confinados (3,01g dL-1±0,43) e a razão A/G (1,07±8,91) foram maiores quando comparados com os bovinos terminados a pasto (2,82g dL-1±0,45) e (0,95±0,38) respectivamente. O nível mais elevado de albumina nos bovinos confinados sugere que eles foram submetidos a uma dieta nutricional mais adequada. O constante desafio imunológico sofrido pelos animais terminados a pasto pode ter sido responsável pelo elevado nível de globulina e fibrinogênio. Esses resultados indicaram que, apesar das adversidades que os bovinos confinados são submetidos, eles não apresentaram alterações...

This research aimed to evaluate the changes in the white blood cell count and some serum proteins of confined cattle (CC) and grass cattle (GC). From the 120 blood samples collected, 60 were obtained from confined Nelore male bovines and 60 from animals with the same characteristics but managed extensively. Samples were obtained at the moment of slaughter. Parameters studied were the white blood cell count, serum albumin/globulin ratio and concentration of plasma fibrinogen. Descriptive statistics was used in the analysis of the data, and the averages, standard deviation and coefficient of variation calculated for all parameters evaluated. The comparisons between averages were made by non-parametric test. The grazing cattle showed higher levels of globulin and fibrinogen when compared to the confined ones (globulin: GC=3.29g dL-1±0.76; CC=2.99g dL-1±0.60 and Fibrinogen: GC=872mg dL-1±610; CC=633mg dL-1±319). The total number of white blood cells mL-1 was 7.64±2.15 in confined cattle and 7.72±1.84 in grazing cattle. There was no significant difference between this variable and differential white blood cell count as well as the total serum protein (g dL-1) from grazing cattle (6.10±0.53) and confined cattle (5.96±0.49). The level of albumin from confined cattle (3.01g dL-1± 0.43) and the A/G ratio (1.07±8.91) were greater when compared to the grazing bovines (2.82g dL-1±0.45) and (0.95±0.38) respectively. The higher serum levels of albumin found in confined herd suggest that they were subjected to a more adequate nutritional diet. The constant immunological challenge suffered by the GC could be responsible for the elevated serum levels of globulin and fibrinogen. These results showed that although feedlots present a stressful environment they did not show any blood alterations correlated to this fact.

Leucograma e teores plasmáticos de proteínas de fase aguda de eqüinos portadores de abdômen agudo e submetidos à laparotomia / Leukogram and plasma acute phase protein concentrations in horses with acute abdomen submitted to treatment by laparotomy

Foram examinados 20 eqüinos adultos portadores de abdômen agudo e submetidos à laparotomia. Dez recuperaram-se sem intercorrência pós-operatória (G1) e 10 foram a óbito sete a 10 dias após a cirurgia, com sinais de choque séptico (G2). Avaliaram-se temperatura retal, freqüências cardíaca e respiratória, tempo de preenchimento capilar e teores plasmáticos das proteínas de fase aguda - fibrinogênio, ceruloplasmina, proteína C-reativa, antitripsina, haptoglobina e glicoproteína ácida -, antes e até sete dias após a laparotomia. As leucometrias às 72h e no sétimo dia pós-operatório dos eqüinos que foram a óbito foram, respectivamente, 34,6 por cento e 57,1 por cento, mais altas que a dos animais curados. Os maiores valores de proteína de fase aguda ocorreram no sétimo dia após a cirurgia; os percentuais de elevação de fibrinogênio, antitripsina, glicoproteina ácida, proteína C-reativa, ceruloplasmina e haptoglobina de eqüinos do G2 em relação ao G1 foram 46,8 por cento, 67,9 por cento, 91,9 por cento, 112,2 por cento, 126,9 por cento e 186,2 por cento, respectivamente.

Twenty adult horses with acute abdomen were examined and submitted to treatment by laparotomy; ten had no postoperative complication (group 1), and ten showed septic shock symptom and died from seven to ten days after surgery (group 2). Body temperature, heart and respiratory rates, filling capillary time, and plasma acute phase protein concentrations - fibrinogen, ceruloplasmin, C-reactive protein, antitrypsin, haptoglobin, and acid glycoprotein - were evaluated before laparotomy and until seven days after surgery. White blood cell counts at 72h and seven days after surgery in group 2 animals were, respectively, 34.6 percent and 57.1 percent, and were higher than those measured in group 1 horses. The highest values of acute phase proteins occurred on the seventh day after surgery. The increase percentages of fibrinogen, antitrypsin, acid glycoprotein, C-reactive protein, ceruloplasmin, and haptoglobin plasma concentrations in group 2 animals in comparison to group 1 animals were 46.8 percent, 67.9 percent, 91.9 percent, 112.2 percent, 126.9 percent, and 186.2 percent, respectively.

Plasma proteomic analysis of patients with squamous cell carcinoma of the uterine cervix / 부인종양

OBJECTIVE: To compare plasma protein expression between patients with squamous cell carcinoma (SCC) of the cervix and normal controls. METHODS: Plasma samples from patients with benign gynecological disease (normal cervix, n=6) and cervical cancer (SCC, n=6) were subjected to plasma proteomic analysis using two dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization mass spectroscopy (MALDI-MS). Western blotting and immunoturbidimetric assay were performed to validate the results of 2-DE. RESULTS: Eight proteins showed differential expression between controls and SCC patients; six (ceruloplasmin, complement C3, afamin precursor, alpha-1-B-glycoprotein, transferrin, alpha-fibrinogen precursor) were up-regulated, while two (chain A, crystal structure of antithrombin and apolipoprotein A-IV precursor) were down-regulated in the plasma of SCC patients. Western blotting analysis revealed significant elevation of ceruloplasmin, complement C3, afamin, and alpha-1-B-glycoprotein in the plasma of SCC patients in comparison to controls. Immunoturbidimetric assay of a larger group confirmed the results of 2-DE and Western blotting, and showed that ceruloplasmin and complement C3 were significantly elevated in the plasma of SCC patients in comparison with controls and patients with carcinoma in situ (CIS) of the uterine cervix. CONCLUSION: Plasma protein expression determined using 2-DE and MALDI-MS will give a chance to identify tumor-specific biomarkers for SCC of the cervix.

An extract of a formula used in the traditional chinese medicine (Buzhong Yi Qi Wan) alters the labeling of blood constituents with technetium-99m

Buzhong Yi Qi Wan (Buzhong) is a medicinal herb widely used in Traditional Chinese Medicine to treat the digestive and circulatory systems. Red blood cell and plasma proteins labeled with technetium-99m (99mTc) are used in nuclear medicine. The aim of this work was to investigate the effects of an aqueous Buzhong extract on the labeling of blood constituents with 99mTc. Heparinized blood (Wistar rats) was incubated in vitro with different Buzhong extract concentrations and 99mTc-labeling was performed. Plasma (P) and blood cells (BC) were separated and soluble (SF-P, SF-BC) and insoluble (IF-P, IF-BC) fractions were isolated. The radioactivity on blood constituents was determined and the percentage of incorporated radioactivity ( percentATI) was calculated. Buzhong extract at the highest concentrations used altered significantly (p<0.05) the percentATI in blood constituents. Substances present in the Buzhong extract could alter the cellular membrane and/or generation of free radicals that have oxidant properties modifying the labeling of blood constituents with 99mTc.

Buzhong Yi Qi Wan (Buzhong) é uma fórmula utilizada na Medicina Tradicional Chinesa para tratamento de distúrbios nos sistemas digestório e circulatório. Constituintes sangüíneos marcados com tecnécio-99m (99mTc) são usados na medicina nuclear. O objetivo deste estudo foi investigar os efeitos do extrato de Buzhong na marcação de constituintes sangüíneos com 99mTc. Amostras de sangue de ratos Wistar foram incubadas com diferentes concentrações do extrato de Buzhong e a marcação de constituintes sangüíneos com 99mTc foi realizado. Plasma e células sangüíneas foram separados, frações solúveis e insolúveis do plasma e das células sangüíneas foram isoladas. A radioatividade nos constituintes sangüíneos foi contada e as porcentagens de radioatividade incorporada ( por centoATI), determinada. Extrato de Buzhong nas maiores concentrações utilizadas altera significativamente (p<0.05) a por centoATI nos constituintes sangüíneos. Substâncias presentes no extrato de Buzhong poderiam alterar a membrana celular e/ou gerar radicais livres, que têm propriedades oxidantes, modificando a marcação dos constituintes sangüíneos com 99mTc.

An aqueous extract of Vitex agnus castus alters the labeling of blood constituents with technetium-99m

The development of experimental assays to study properties of herbal medicine is worthwhile. Vitex agnus castus (VAC) is utilized in popular medicine and some actions have been attributed to its extract. Blood cells (BC) and plasma proteins are labeled with technetium-99m (Tc-99m) and have been used in nuclear medicine, as in basic research. This procedure uses a reducing agent and stannous ion is utilized. There are reports that drugs can alter this labeling process. The aim of this work was to evaluate the influence of an aqueous extract of VAC on the labeling of blood constituents with Tc-99m. Blood was incubated with VAC, stannous chloride and Tc-99m, as sodium pertechnetate, and centrifuged. Samples of BC and plasma were separated, aliquots of BC and plasma were also precipitated with trichloroacetic acid to obtain soluble and insoluble fractions and the percentage of radioactivity ( percentATI) was determined. The results show a statistical (p<0.05) alteration in the percentATI on blood compartments and on the insoluble fractions of plasma and BC. Probably, this extract would have chemical compounds with oxidant properties.

Modelos experimentais são relevantes no estudo de propriedades de plantas medicinais. Vitex agnus castus (VAC) é usado na medicina popular. Células sanguíneas (CS) e proteínas plasmáticas são marcadas com tecnécio-99m (Tc-99m) com aplicações na medicina nuclear e em pesquisa. Esse procedimento utiliza um agente redutor e o íon estanoso é usado. Drogas podem alterar esse processo de marcação. O objetivo desse trabalho foi avaliar a influência de um extrato aquoso de VAC na marcação de constituintes sanguíneos com Tc-99m. Sangue foi incubado com VAC, cloreto estanoso e Tc-99m, como pertecnetato de sódio e centrifugado. Amostras de CS e plasma foram separadas, alíquotas de CS e plasma foram também precipitadas com ácido tricloroacético para obtenção de frações solúvel (FS) e insolúvel (FI) e a percentagem de radioatividade ( por centoATI) foi determinada. Os resultados mostraram uma alteração estatística (p<0.05) na por centoATI dos compartimentos sanguíneos e nas FI do plasma e CS. Provavelmente, esse extrato poderia ter compostos químicos com propriedades oxidantes.

An aqueous extract of Pfaffia sp. does not alter the labeling of blood constituents with technetium-99m and the morphology of the red blood cells

Plants have been used for the human beings as food, as additives and/or as medicines. There are controversies about the biological effects of several natural products and, it is worthwhile to try to develop experimental assays to evaluate properties of extracts of plants. Pfaffia sp. is utilized in popular medicine and various properties have been attributed to its extract. Red blood cells (RBC) and plasma proteins are labeled with technetium-99m (Tc-99m) and this labeling procedure depends on a reducing agent and stannous ion is usually used. There are reports that drugs can alter the labeling of blood elements with Tc-99m. We have evaluated the influence of a Pfaffia sp. extract on the labeling of blood constituents with Tc-99m and on the morphology of RBC. Blood was incubated with an aqueous extract of Pfaffia sp., stannous chloride and Tc-99m. Samples were centrifuged and plasma and blood cells were separated and also precipitated with trichloroacetic acid. Soluble and insoluble fractions were separated. The results did not show alteration in the uptake of radioactivity and no modifications on the shape of the RBC in presence of Pfaffia sp. Once this labeling process depends on a reducing agent, probably, this extract has compounds with anti-oxidant properties as already described elsewhere, that could protect the stannous ions against the oxidation process. This fact would aid the labeling process of blood elements with Tc-99m.

Inhibition of Pulmonary Surfactant Function by Endotoxic Rat Plasma and Its Mechanisms / 第二军医大学学报

The inhibition of bronchoalveolar surfactant function by endotoxic rat plasma was compared with that by normal rat plasma with M-8601 film balance. The results showed that endotoxic rat plasma significantly inhibited surfactant function of bronchoalveolar lavage (BAL). In the same protein concentration, the inhibition effects caused by endotoxic rat plasma were comparable with those by normal rat plasma. There were no changes of the inhibition effects after incubation of BAL with either endotoxic or normal plasma at 37℃ for 30min. These results suggest that the inhibition of surfactant function by the leakage of plasma in endotoxic lung injury is mainly due to protein components.