Cell therapy in dilated cardiomyopathy: from animal models to clinical trials

Dilated cardiomyopathy can be the end-stage form and common denominator of several cardiac disorders of known cause, such as hypertensive, ischemic, diabetic and Chagasic diseases. However, some individuals have clinical findings, such as an increase in ventricular chamber size and impaired contractility (classical manifestations of dilated cardiomyopathy) even in the absence of a diagnosed primary disease. In these patients, dilated cardiomyopathy is classified as idiopathic since its etiology is obscure. Nevertheless, regardless of all of the advances in medical, pharmacological and surgical procedures, the fate of patients with dilated cardiomyopathy (of idiopathic or of any other known cause) is linked to arrhythmic episodes, severe congestive heart failure and an increased risk of sudden cardiac death. In this review, we will summarize present data on the use of cell therapies in animal models of dilated cardiomyopathies and will discuss the few clinical trials that have been published so far involving patients affected by this disease. The animal models discussed here include those in which the cardiomyopathy is produced by genetic manipulation and those in which disease is induced by chemical or infectious agents. The specific model used clearly creates restrictions to translation of the proposed cell therapy to clinical practice, insofar as most of the clinical trials performed to date with cell therapy have used autologous cells. Thus, translation of genetic models of dilated cardiomyopathy may have to wait until the use of allogeneic cells becomes more widespread in clinical trials of cell therapies for cardiac diseases.

Percutaneous sciatic nerve block with tramadol induces analgesia and motor blockade in two animal pain models

Local anesthetic efficacy of tramadol has been reported following intradermal application. Our aim was to investigate the effect of perineural tramadol as the sole analgesic in two pain models. Male Wistar rats (280-380 g; N = 5/group) were used in these experiments. A neurostimulation-guided sciatic nerve block was performed and 2% lidocaine or tramadol (1.25 and 5 mg) was perineurally injected in two different animal pain models. In the flinching behavior test, the number of flinches was evaluated and in the plantar incision model, mechanical and heat thresholds were measured. Motor effects of lidocaine and tramadol were quantified and a motor block score elaborated. Tramadol, 1.25 mg, completely blocked the first and reduced the second phase of the flinching behavior test. In the plantar incision model, tramadol (1.25 mg) increased both paw withdrawal latency in response to radiant heat (8.3 ± 1.1, 12.7 ± 1.8, 8.4 ± 0.8, and 11.1 ± 3.3 s) and mechanical threshold in response to von Frey filaments (459 ± 82.8, 447.5 ± 91.7, 320.1 ± 120, 126.43 ± 92.8 mN) at 5, 15, 30, and 60 min, respectively. Sham block or contralateral sciatic nerve block did not differ from perineural saline injection throughout the study in either model. The effect of tramadol was not antagonized by intraperitoneal naloxone. High dose tramadol (5 mg) blocked motor function as well as 2% lidocaine. In conclusion, tramadol blocks nociception and motor function in vivo similar to local anesthetics.

Animal models of prenatal immune challenge and their contribution to the study of schizophrenia: a systematic review

Prenatal immune challenge (PIC) in pregnant rodents produces offspring with abnormalities in behavior, histology, and gene expression that are reminiscent of schizophrenia and autism. Based on this, the goal of this article was to review the main contributions of PIC models, especially the one using the viral-mimetic particle polyriboinosinic-polyribocytidylic acid (poly-I:C), to the understanding of the etiology, biological basis and treatment of schizophrenia. This systematic review consisted of a search of available web databases (PubMed, SciELO, LILACS, PsycINFO, and ISI Web of Knowledge) for original studies published in the last 10 years (May 2001 to October 2011) concerning animal models of PIC, focusing on those using poly-I:C. The results showed that the PIC model with poly-I:C is able to mimic the prodrome and both the positive and negative/cognitive dimensions of schizophrenia, depending on the specific gestation time window of the immune challenge. The model resembles the neurobiology and etiology of schizophrenia and has good predictive value. In conclusion, this model is a robust tool for the identification of novel molecular targets during prenatal life, adolescence and adulthood that might contribute to the development of preventive and/or treatment strategies (targeting specific symptoms, i.e., positive or negative/cognitive) for this devastating mental disorder, also presenting biosafety as compared to viral infection models. One limitation of this model is the incapacity to model the full spectrum of immune responses normally induced by viral exposure.

Performance assessment of the single photon emission microscope: high spatial resolution SPECT imaging of small animal organs

The single photon emission microscope (SPEM) is an instrument developed to obtain high spatial resolution single photon emission computed tomography (SPECT) images of small structures inside the mouse brain. SPEM consists of two independent imaging devices, which combine a multipinhole collimator, a high-resolution, thallium-doped cesium iodide [CsI(Tl)] columnar scintillator, a demagnifying/intensifier tube, and an electron-multiplying charge-coupling device (CCD). Collimators have 300- and 450-µm diameter pinholes on tungsten slabs, in hexagonal arrays of 19 and 7 holes. Projection data are acquired in a photon-counting strategy, where CCD frames are stored at 50 frames per second, with a radius of rotation of 35 mm and magnification factor of one. The image reconstruction software tool is based on the maximum likelihood algorithm. Our aim was to evaluate the spatial resolution and sensitivity attainable with the seven-pinhole imaging device, together with the linearity for quantification on the tomographic images, and to test the instrument in obtaining tomographic images of different mouse organs. A spatial resolution better than 500 µm and a sensitivity of 21.6 counts·s-1·MBq-1 were reached, as well as a correlation coefficient between activity and intensity better than 0.99, when imaging 99mTc sources. Images of the thyroid, heart, lungs, and bones of mice were registered using 99mTc-labeled radiopharmaceuticals in times appropriate for routine preclinical experimentation of <1 h per projection data set. Detailed experimental protocols and images of the aforementioned organs are shown. We plan to extend the instrument's field of view to fix larger animals and to combine data from both detectors to reduce the acquisition time or applied activity.

Laboratory animal: biological reagent or living being?

The duties of humans toward non-human animals and their rights in society have been debated for a long time. However, a discussion on the terminology used for the identification of laboratory animals is usually not considered, although the employment of inadequate terminology may generate disastrous consequences for the animals before, during, and after the experiment. This study intends to defend the use of appropriate terminology, call attention to an unethical attitude of certain professionals when dealing with experimental animals, and also propose operational mechanisms, which allow for those distortions to be corrected.

CIAPIN1 gene silencing enhances chemosensitivity in a drug-resistant animal model in vivo

Overexpression of cytokine-induced apoptosis inhibitor 1 (CIAPIN1) contributes to multidrug resistance (MDR) in breast cancer. This study aimed to evaluate the potential of CIAPIN1 gene silencing by RNA interference (RNAi) as a treatment for drug-resistant breast cancer and to investigate the effect of CIAPIN1 on the drug resistance of breast cancer in vivo. We used lentivirus-vector-based RNAi to knock down CIAPIN1 in nude mice bearing MDR breast cancer tumors and found that lentivirus-vector-mediated silencing of CIAPIN1 could efficiently and significantly inhibit tumor growth when combined with chemotherapy in vivo. Furthermore, Western blot analysis showed that both CIAPIN1 and P-glycoprotein expression were efficiently downregulated, and P53 was upregulated, after RNAi. Therefore, we concluded that lentivirus-vector-mediated RNAi targeting of CIAPIN1 is a potential approach to reverse MDR of breast cancer. In addition, CIAPIN1 may participate in MDR of breast cancer by regulating P-glycoprotein and P53 expression.

An animal experimental study of porous magnesium scaffold degradation and osteogenesis

Our objective was to observe the biodegradable and osteogenic properties of magnesium scaffolding under in vivo conditions. Twelve 6-month-old male New Zealand white rabbits were randomly divided into two groups. The chosen operation site was the femoral condyle on the right side. The experimental group was implanted with porous magnesium scaffolds, while the control group was implanted with hydroxyapatite scaffolds. X-ray and blood tests, which included serum magnesium, alanine aminotransferase (ALT), creatinine (CREA), and blood urea nitrogen (BUN) were performed serially at 1, 2, and 3 weeks, and 1, 2, and 3 months. All rabbits were killed 3 months postoperatively, and the heart, kidney, spleen, and liver were analyzed with hematoxylin and eosin (HE) staining. The bone samples were subjected to microcomputed tomography scanning (micro-CT) and hard tissue biopsy. SPSS 13.0 (USA) was used for data analysis, and values of P<0.05 were considered to be significant. Bubbles appeared in the X-ray of the experimental group after 2 weeks, whereas there was no gas in the control group. There were no statistical differences for the serum magnesium concentrations, ALT, BUN, and CREA between the two groups (P>0.05). All HE-stained slices were normal, which suggested good biocompatibility of the scaffold. Micro-CT showed that magnesium scaffolds degraded mainly from the outside to inside, and new bone was ingrown following the degradation of magnesium scaffolds. The hydroxyapatite scaffold was not degraded and had fewer osteoblasts scattered on its surface. There was a significant difference in the new bone formation and scaffold bioabsorption between the two groups (9.29±1.27 vs 1.40±0.49 and 7.80±0.50 vs 0.00±0.00 mm3, respectively; P<0.05). The magnesium scaffold performed well in degradation and osteogenesis, and is a promising material for orthopedics.

A forced running wheel system with a microcontroller that provides high-intensity exercise training in an animal ischemic stroke model

We developed a forced non-electric-shock running wheel (FNESRW) system that provides rats with high-intensity exercise training using automatic exercise training patterns that are controlled by a microcontroller. The proposed system successfully makes a breakthrough in the traditional motorized running wheel to allow rats to perform high-intensity training and to enable comparisons with the treadmill at the same exercise intensity without any electric shock. A polyvinyl chloride runway with a rough rubber surface was coated on the periphery of the wheel so as to permit automatic acceleration training, and which allowed the rats to run consistently at high speeds (30 m/min for 1 h). An animal ischemic stroke model was used to validate the proposed system. FNESRW, treadmill, control, and sham groups were studied. The FNESRW and treadmill groups underwent 3 weeks of endurance running training. After 3 weeks, the experiments of middle cerebral artery occlusion, the modified neurological severity score (mNSS), an inclined plane test, and triphenyltetrazolium chloride were performed to evaluate the effectiveness of the proposed platform. The proposed platform showed that enhancement of motor function, mNSS, and infarct volumes was significantly stronger in the FNESRW group than the control group (P<0.05) and similar to the treadmill group. The experimental data demonstrated that the proposed platform can be applied to test the benefit of exercise-preconditioning-induced neuroprotection using the animal stroke model. Additional advantages of the FNESRW system include stand-alone capability, independence of subjective human adjustment, and ease of use.

Ocorrência de Vibrio vulnificus em alguns alimentos de origem marinha

Vibrio vulnificus é uma bactéria Gram-negativa que habita águas marinhas. É patogênica para o homem e a doença está associada ao consumo de frutos do mar, com aproximadamente 60% dos casos sendo fatais em pacientes imunocomprometidos. O objetivo desta pesquisa foi estudar a ocorrência de V. vulnificus em amostras de alguns alimentos de origem marinha. As amostras de ostras, mariscos e camarões foram coletadas nos períodos de abril-agosto de 1993, maio-setembro de 1994 e fevereiro de 1995. De 55 amostras de ostras, 36 (65%) foram positivas para V. vulnificus; das 19 amostras de mariscos, 8 (42,1%) foram positivas e das 7 amostras de camarão, 1 (4,3%) foi positiva para esta bactéria. Os resultados permitiram-nos concluir que a bactéria foi recuperada durante todos os meses de análise, demonstrando que estes alimentos, principalmente quando consumidos crus, são potencialmente perigosos para os seres humanos na faixa de risco.

NOVAS APLICAÇÕES DE SISTEMAS SFE "HOME-MADE".: IV. QUALIDADE DOS ALIMENTOS

São apresentadas algumas aplicações de um sistema "Home-Made" para Extração com Fluido Supercrítico (EFS) na análise de resíduos de pesticidas e de óleos essenciais em alimentos. Foram desenvolvidos diversos estudos com os parâmetros: temperatura, pressão, escolha e concentração de modificadores. Os resultados são apresentados para amostras de maracujá (resíduos de pesticidas organofosforados e piretróides), mel (resíduos de pesticidas organoclorados) e laranja (óleo essencial).

CARACTERÍSTICAS DE QUALIDADE DO CONTRAFILÉ (m. L. dorsi) DE TOUROS JOVENS DA RAÇA NELORE

Com o objetivo de analisar as características de qualidade da carne de touros jovens da raça Nelore e os possíveis efeitos da idade, entre os 690 e os 780 dias, nessas características, efetuou-se o abate de 113 bovinos, que haviam sido confinados por um período de 109 dias com ração à base de 20% de concentrado e 80% de volumoso. Todas as carcaças foram tipificadas pela Inspeção Federal na sala de matança do frigorífico e resfriadas por 24 h em câmara fria (Tinicial=5°C; Tfinal=2°C). Meias carcaças (lado direito, N=51), do tipo B - do Sistema B R A S I L - de animais de 23 a 26 meses foram desossadas. Dois bifes (2,5cm de espessura) de contrafilé (m. L. dorsi) de cada uma foram embalados à vácuo e maturados por 7 dias a 0-2°C. O pH variou entre 5,44 e 5,83 e apenas duas amostras tinham pH ³ 5,70. O valor L* (luminosidade) médio foi de 34,85. As médias de umidade e gordura foram de 75,65% e 1,71%, respectivamente. A média de força de cisalhamento foi de 6,70kg e não foi influenciada pela idade do animal entre 690 e 734 dias, mas apresentou uma tendência (Teste t, p=0,22) de aumento entre 735 e 780 dias. Não se detectou influência da idade do animal nas demais características de qualidade analisadas (Teste t, p>0,20). Concluiu-se que o contrafilé de animais como esses pode ter problemas de aceitação na faixa mais exigente de mercado, e que carcaças tipificadas como B, presumivelmente o melhor tipo, não apresentam necessariamente as melhores características de qualidade da carne.

COMPARAÇÃO ENTRE TÉCNICAS NUMÉRICAS PARA A RESOLUÇÃO DO PROBLEMA DE TRANSFERÊNCIA DE CALOR EM ALIMENTOS ENLATADOS

Este trabalho apresenta a comparação das técnicas de resolução numérica de Diferenças Finitas e Runge-Kutta-Gill de 4a ordem com passo de integração variável para a simulação do modelo de transferência de calor em regime transiente bidimensional em coordenadas cilíndricas e unidimensional em coordenadas esféricas, aplicado a alimentos enlatados processados em autoclave, observando-se a rapidez e a precisão de integração comparados aos perfis reais de tempo/temperatura, incluindo desvios de processo. Além disso, uma análise de sensibilidade paramétrica na difusividade térmica foi realizada na tentativa de quantificar a influência que essa propriedade possui na resolução numérica. Para tanto, foram realizados ensaios de penetração de calor em autoclave a vapor, utilizando-se latas cilíndricas de 75x90 e 100x110 contendo simulantes de alimentos com características de aquecimento por condução e convecção.

Características físico-químicas dos méis do município de São Fidélis-RJ

De acordo com a tonalidade as amostras de mel foram divididas em grupos, armazenadas em diferentes condições e analisadas segundo os métodos adotados pelo Instituto Adolfo Lutz, Laboratório Nacional de Referência Animal e Association of Official Analytical Chemists. As amostras foram acondicionadas em recipientes de vidro e de polipropileno e armazenadas à temperatura ambiente. Um grupo de amostras foi armazenada em luminosidade ambiente enquanto o outro grupo ao abrigo da luz. As determinações de umidade, pH, acidez, cor, açúcares redutores e não-redutores, hidroximetilfurfural e índices de Lugol, Lund, Fiehe e diastase foram efetuadas por um período de um ano de armazenamento. Os resultados obtidos indicam que as amostras analisadas mantiveram suas características dentre os padrões aceitáveis e que os produtores devem ser orientados para o uso de técnicas de acondicionamento e armazenamento dos méis, para garantir a qualidade dos seus produtos.

Efecto del fraccionamento sobre las caracteristicas del germen de maiz desgrasado

El germen de maíz desgrasado (GMD) es un subproducto que se obtiene al elaborar harina cruda o precocida y extraer el aceite de maíz. Este subproducto se usa exclusivamente en la alimentación animal, debido a que es un material heterogéneo con alto contenido de salvado. El principal obstáculo para usarlo en la formulación de alimentos es que los fragmentos de fibra de la cáscara, modifican las características sensoriales y funcionales de los productos. Para evitar esta limitante, en este trabajo, se trató de eliminar los fragmentos de fibra y de incrementar la concentración de proteína. Para esto, el material extraíble (ME) rico en grasa y el germen industrial desgrasado (GD) fueron sometidos a fraccionamientos mediante separación en tamices, aspirado y molienda. El rendimiento en fracciones de GMD obtenidas a nivel de laboratorio GD1 e industrial GD2 respectivamente, fue de 44,76% y 78,02%. La fibra se redujo en un 18,45% y en un 41,61% y se aumentó la proteína en 7,40% y en 19,56% para las fracciones GD1 y GD2 respectivamente. La calidad de la proteína medida por la eficiencia proteica (PER) varió de 2,57 a 2,84 lo que resultó ser 16 veces superior a la de la harina de endospermo de maíz blanco (PER 0,16) y la digestibilidad aparente de las fracciones GD1 y GD2 dio valores superiores al 86%, lo que indica que el fraccionamiento es una tecnología sencilla que mejora considerablemente la calidad de las proteínas del germen, el color y los índices de solubilidad y absorción de agua.

Estudo da biomassa de aguapé, para a produção do seu concentrado protéico

Na produção de concentrado protéico de aguapé (C. P. A.), utilizou-se para extração da fração protéica solução de hidróxido de sódio (50mM) na proporção (3:1) sendo três volumes de solução de hidróxido de sódio para um volume de planta desintegrada. A precipitação da proteína foi realizada utilizando solução de ácido clorídrico (100mM) à temperatura de 80° C, o pH foi sendo corrigido até atingir o ponto isoelétrico da proteína, que ocorreu em pH 3,5. O teor de proteína encontrado no C. P. A. foi superior ao encontrado em outros vegetais e produtos de origem vegetal tais como alfafa e farinha de soja por exemplo. A presença de todos aminoácidos essenciais no C. P. A. mostra que sua utilização é viável na produção não só de ração animal, mas também, como suplementação alimentar na dieta humana.