Effect of oral ingestion of an extract of the herb Uncaria tomentosa on the biodistribution of sodium pertechnetate in rats

The aim of the present study was to determine the effect of the oral ingestion of an extract of the herb Uncaria tomentosa (cat's claw) on the biodistribution of the radiobiocomplex sodium pertechnetate (Na99mTcO4) in rats. The animals (male Wistar rats, 2 months old, 180-220 g), were treated (1 mL) with an U. tomentosa extract (32 mg/mL, N = 5) or 0.9% NaCl solution (control, N = 5) for 7 days. After this period, Na99mTcO4 (3.7 MBq, 0.3 mL) was injected through the ocular plexus and after 10 min the rats were killed, the organs isolated and counted in a well-gamma counter. A significant (P < 0.05) alteration in Na99mTcO4 uptake i) from 0.57 ± 0.008 to 0.39 ± 0.06 %ATI/organ (P < 0.05) and from 0.57 ± 0.17 to 0.39 ± 0.14 %ATI/g (P < 0.05) was observed in the heart, ii) from 0.07 ± 0.02 to 0.19 ± 0.07 %ATI/g in the pancreas, and iii) from 0.07 ± 0.01 to 0.18 ± 0.07 %ATI/g (P < 0.05) in muscle after treatment with this extract. Although these results were obtained with animals, caution is advisable in the interpretation of the nuclear medicine examination when the patient is using this herb. This finding is probably an example of drug interaction with a radiopharmaceutical, a fact that could lead to misdiagnosis of the examination in clinical practice with unexpected consequences for the patient.

Long-term aerobic swimming training by rats reduces the number of aberrant crypt foci in 1,2-dimethylhydrazine-induced colon cancer

We determined the effect of long-term aerobic swimming training regimens of different intensities on colonic carcinogenesis in rats. Male Wistar rats (11 weeks old) were given 4 subcutaneous injections (40 mg/kg body weight each) of 1,2-dimethyl-hydrazine (DMH, dissolved in 0.9% NaCl containing 1.5% EDTA, pH 6.5), at 3-day intervals and divided into three exercise groups that swam with 0% body weight (EG1, N = 11), 2% body weight (EG2, N = 11), and 4% body weight of load (EG3, N = 10), 20 min/day, 5 days/week for 35 weeks, and one sedentary control group (CG, N = 10). At sacrifice, the colon was removed and counted for tumors and aberrant crypt foci. Tumor size was measured and intra-abdominal fat was weighed. The mean number of aberrant crypt foci was reduced only for EG2 compared to CG (26.21 ± 2.99 vs 36.40 ± 1.53 crypts; P < 0.05). Tumor incidence was not significantly different among groups (CG: 90%; EG1: 72.7%; EG2: 90%; EG3: 80%). Swimming training did not affect either tumor multiplicity (CG: 2.30 ± 0.58; EG1: 2.09 ± 0.44; EG2: 1.27 ± 0.19; EG3: 1.50 ± 0.48 tumors) or size (CG: 1.78 ± 0.24; EG1: 1.81 ± 0.14; EG2: 1.55 ± 0.21; EG3: 2.17 ± 0.22 cm³). Intra-abdominal fat was not significantly different among groups (CG: 10.54 ± 2.73; EG1: 6.12 ± 1.15; EG2: 7.85 ± 1.24; EG3: 5.11 ± 0.74 g). Aerobic swimming training with 2% body weight of load protected against the DMH-induced preneoplastic colon lesions, but not against tumor development in the rat.

Treatment of hemorrhagic shock with hypertonic saline solution modulates the inflammatory response to live bacteria in lungs

Shock and resuscitation render patients more susceptible to acute lung injury due to an exacerbated immune response to subsequent inflammatory stimuli. To study the role of innate immunity in this situation, we investigated acute lung injury in an experimental model of ischemia-reperfusion (I-R) followed by an early challenge with live bacteria. Conscious rats (N = 8 in each group) were submitted to controlled hemorrhage and resuscitated with isotonic saline (SS, 0.9% NaCl) or hypertonic saline (HS, 7.5% NaCl) solution, followed by intratracheal or intraperitoneal inoculation of Escherichia coli. After infection, toll-like receptor (TLR) 2 and 4 mRNA expression was monitored by RT-PCR in infected tissues. Plasma levels of tumor necrosis factor α and interleukins 6 and 10 were determined by ELISA. All animals showed similar hemodynamic variables, with mean arterial pressure decreasing to nearly 40 mmHg after bleeding. HS or SS used as resuscitation fluid yielded equal hemodynamic results. Intratracheal E. coli inoculation per se induced a marked neutrophil infiltration in septa and inside the alveoli, while intraperitoneal inoculation-associated neutrophils and edema were restricted to the interseptal space. Previous I-R enhanced lung neutrophil infiltration upon bacterial challenge when SS was used as reperfusion fluid, whereas neutrophil influx was unchanged in HS-treated animals. No difference in TLR expression or cytokine secretion was detected between groups receiving HS or SS. We conclude that HS is effective in reducing the early inflammatory response to infection after I-R, and that this phenomenon is achieved by modulation of factors other than expression of innate immunity components.

Antipyretic and antioxidant activities of 5-trifluoromethyl-4,5-dihydro-1H-pyrazoles in rats

The objective of this study was to determine the effect of eight 5-hydroxy-5-trifluoromethyl-4,5-dihydro-1H-1-carboxyamidepyrazoles (TFDPs) on rat body temperature and baker’s yeast-induced fever. TFDPs or vehicle (5% Tween 80 in 0.9% NaCl, 5 mL/kg) were injected subcutaneously and rectal temperature was measured as a function of time in 28-day-old male Wistar rats (N = 5-12 per group). Antipyretic activity was determined in feverish animals injected with baker’s yeast (Saccharomyces cerevisiae suspension, 0.135 mg/kg, 10 mL/kg, ip). 3-Ethyl- and 3-propyl-TFDP (140 and 200 μmol/kg, respectively, 4 h after yeast injection) attenuated baker’s yeast-induced fever by 61 and 82%, respectively. These two effective antipyretics were selected for subsequent analysis of putative mechanisms of action. We then determined the effects on cyclooxygenase-1 and -2 (COX-1 and COX-2) activities on 1,1-diphenyl-2-picrylhydrazyl (DPPH) oxidation in vitro, on tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels and on leukocyte counts in the washes of peritoneal cavities of rats injected with baker’s yeast. While 3-ethyl- and 3-propyl-TFDP did not reduce baker’s yeast-induced increases of IL-1β or TNF-α levels, 3-ethyl-TFDP caused a 42% reduction in peritoneal leukocyte count. 3-Ethyl- and 3-propyl-TFDP did not alter COX-1 or COX-2 activities in vitro, but presented antioxidant activity in the DPPH assay with an IC50 of 39 mM (25-62) and 163 mM (136-196), respectively. The data indicate that mechanisms of action of these two novel antipyretic pyrazole derivatives do not involve the classic inhibition of the COX pathway or pyrogenic cytokine release.

Long-term melatonin treatment reduces ovarian mass and enhances tissue antioxidant defenses during ovulation in the rat

Melatonin regulates the reproductive cycle, energy metabolism and may also act as a potential antioxidant indoleamine. The present study was undertaken to investigate whether long-term melatonin treatment can induce reproductive alterations and if it can protect ovarian tissue against lipid peroxidation during ovulation. Twenty-four adult female Wistar rats, 60 days old (± 250-260 g), were randomly divided into two equal groups. The control group received 0.3 mL 0.9% NaCl + 0.04 mL 95% ethanol as vehicle, and the melatonin-treated group received vehicle + melatonin (100 µg·100 g body weight-1·day-1) both intraperitoneally daily for 60 days. All animals were killed by decapitation during the morning estrus at 4:00 am. Body weight gain and body mass index were reduced by melatonin after 10 days of treatment (P < 0.05). Also, a marked loss of appetite was observed with a fall in food intake, energy intake (melatonin 51.41 ± 1.28 vs control 57.35 ± 1.34 kcal/day) and glucose levels (melatonin 80.3 ± 4.49 vs control 103.5 ± 5.47 mg/dL) towards the end of treatment. Melatonin itself and changes in energy balance promoted reductions in ovarian mass (20.2%) and estrous cycle remained extensive (26.7%), arresting at diestrus. Regarding the oxidative profile, lipid hydroperoxide levels decreased after melatonin treatment (6.9%) and total antioxidant substances were enhanced within the ovaries (23.9%). Additionally, melatonin increased superoxide dismutase (21.3%), catalase (23.6%) and glutathione-reductase (14.8%) activities and the reducing power (10.2% GSH/GSSG ratio). We suggest that melatonin alters ovarian mass and estrous cyclicity and protects the ovaries by increasing superoxide dismutase, catalase and glutathione-reductase activities.

Effects of simultaneous exposure to stress and nicotine on nicotine-induced locomotor activation in adolescent and adult rats

Preclinical studies have shown that repeated stress experiences can result in an increase in the locomotor response to the subsequent administration of drugs of abuse, a phenomenon that has been termed behavioral cross-sensitization. Behavioral sensitization reflects neuroadaptive processes associated with drug addiction and drug-induced psychosis. Although cross-sensitization between stress- and drug-induced locomotor activity has been clearly demonstrated in adult rats, few studies have evaluated this phenomenon in adolescent rats. In the present study, we determined if the simultaneous exposure to stress and nicotine was capable of inducing behavioral sensitization to nicotine in adolescent and adult rats. To this end, adolescent (postnatal day (P) 28-37) and adult (P60-67) rats received nicotine (0.4 mg/kg, sc) or saline (0.9% NaCl, sc) and were immediately subjected to restraint stress for 2 h once a day for 7 days. The control group for stress was undisturbed following nicotine or saline injections. Three days after the last exposure to stress and nicotine, rats were challenged with a single dose of nicotine (0.4 mg/kg, sc) or saline and nicotine-induced locomotion was then recorded for 30 min. In adolescent rats, nicotine caused behavioral sensitization only in animals that were simultaneously exposed to stress, while in adult rats nicotine promoted sensitization independently of stress exposure. These findings demonstrate that adolescent rats are more vulnerable to the effects of stress on behavioral sensitization to nicotine than adult rats.

Transplantation of mononuclear cells from human umbilical cord blood promotes functional recovery after traumatic spinal cord injury in Wistar rats

Cell transplantation is a promising experimental treatment for spinal cord injury. The aim of the present study was to evaluate the efficacy of mononuclear cells from human umbilical cord blood in promoting functional recovery when transplanted after a contusion spinal cord injury. Female Wistar rats (12 weeks old) were submitted to spinal injury with a MASCIS impactor and divided into 4 groups: control, surgical control, spinal cord injury, and one cell-treated lesion group. Mononuclear cells from umbilical cord blood of human male neonates were transplanted in two experiments: a) 1 h after surgery, into the injury site at a concentration of 5 x 10(6) cells diluted in 10 µL 0.9% NaCl (N = 8-10 per group); b) into the cisterna magna, 9 days after lesion at a concentration of 5 x 10(6) cells diluted in 150 µL 0.9% NaCl (N = 12-14 per group). The transplanted animals were immunosuppressed with cyclosporin-A (10 mg/kg per day). The BBB scale was used to evaluate motor behavior and the injury site was analyzed with immunofluorescent markers to label human transplanted cells, oligodendrocytes, neurons, and astrocytes. Spinal cord injury rats had 25% loss of cord tissue and cell treatment did not affect lesion extension. Transplanted cells survived in the injured area for 6 weeks after the procedure and both transplanted groups showed better motor recovery than the untreated ones (P < 0.05). The transplantation of mononuclear cells from human umbilical cord blood promoted functional recovery with no evidence of cell differentiation.

Panic-like defensive behavior but not fear-induced antinociception is differently organized by dorsomedial and posterior hypothalamic nuclei of Rattus norvegicus (Rodentia, Muridae)

The hypothalamus is a forebrain structure critically involved in the organization of defensive responses to aversive stimuli. Gamma-aminobutyric acid (GABA)ergic dysfunction in dorsomedial and posterior hypothalamic nuclei is implicated in the origin of panic-like defensive behavior, as well as in pain modulation. The present study was conducted to test the difference between these two hypothalamic nuclei regarding defensive and antinociceptive mechanisms. Thus, the GABA A antagonist bicuculline (40 ng/0.2 µL) or saline (0.9% NaCl) was microinjected into the dorsomedial or posterior hypothalamus in independent groups. Innate fear-induced responses characterized by defensive attention, defensive immobility and elaborate escape behavior were evoked by hypothalamic blockade of GABA A receptors. Fear-induced defensive behavior organized by the posterior hypothalamus was more intense than that organized by dorsomedial hypothalamic nuclei. Escape behavior elicited by GABA A receptor blockade in both the dorsomedial and posterior hypothalamus was followed by an increase in nociceptive threshold. Interestingly, there was no difference in the intensity or in the duration of fear-induced antinociception shown by each hypothalamic division presently investigated. The present study showed that GABAergic dysfunction in nuclei of both the dorsomedial and posterior hypothalamus elicit panic attack-like defensive responses followed by fear-induced antinociception, although the innate fear-induced behavior originates differently in the posterior hypothalamus in comparison to the activity of medial hypothalamic subdivisions.

Short-term levosimendan treatment protects rat testes against oxidative stress

The objective of this study was to evaluate the effect of short-term levosimendan exposure on oxidant/antioxidant status and trace element levels in the testes of rats under physiological conditions. Twenty male Wistar albino rats were randomly divided into two groups of 10 animals each. Group 1 was not exposed to levosimendan and served as control. Levosimendan (12 µg/kg) diluted in 10 mL 0.9% NaCl was administered intraperitoneally to group 2. Animals of both groups were sacrificed after 3 days and their testes were harvested for the determination of changes in tissue oxidant/antioxidant status and trace element levels. Tissue malondialdehyde (MDA) was significantly lower in the levosimendan group (P < 0.001) than in the untreated control group and superoxide dismutase and glutathione peroxidase (GSH-Px) levels were significantly higher in the levosimendan group (P < 0.001). Carbonic anhydrase, catalase and GSH levels were not significantly different from controls. Mg and Zn levels of testes were significantly higher (P < 0.001) and Co, Pb, Cd, Mn, and Cu were significantly lower (P < 0.001) in group 2 compared to group 1. Fe levels were similar for the two groups (P = 0.94). These results suggest that 3-day exposure to levosimendan induced a significant decrease in tissue MDA level, which is a lipid peroxidation product and an indicator of oxidative stress, and a significant increase in the activity of an important number of the enzymes that protect against oxidative stress in rat testes.

Determinação dos teores de cobre em diferentes extratos de hortaliças do tipo A e B

Amostras de hortaliças dos tipos A e B foram analisadas quanto ao teor total de cobre através de espectroscopia de absorção atômica em chama, e o método de extração seqüencial foi aplicado com os seguintes extratores: solução de CaCl(2)1,0M; solução de ácido acético 0,1M; solução de ácido acético 0,5M / acetato de amônio 5% [pH=5,0]; solução de NaOH 0,1M e solução de HCl 0,5M. Na análise da rejeição de resultados foi aplicado o teste de Grubbs e na comparação dos dados o teste t de Student foi utilizado. Em média 25,4% do teor total de cobre foram extraídos com solução de CaCl2 1,0M, sendo o máximo de 50,5% e o mínimo de 8,3%. Em média 14,8% do teor total de cobre, sendo que 32,6% e 6,4%, máximo e mínimo respectivamente, foram extraídos com solução de ácido acético 0,1M. Com relação ao uso de solução de ácido acético 0,5M / acetato de amônio 5% [pH=5,0], o mínimo extraído obteve valor de 6,2% e o máximo 27,6%, com média de 13,7%. O extrator de NaOH 0,1M foi o que teve menor extração, cerca de 10,5% e com extrator de HCl 0,5M a extração foi em média de 12,6%. A fração extraída de cobre nas amostras foi de no mínimo 34,5% e no máximo 100%. Observa-se que para a maioria das amostras, o cobre se encontra sob a forma de, no mínimo, 6 espécies químicas distintas.

Determinação dos teores de zinco em diferentes extratos de hortaliças dos tipos A e B

O zinco é um elemento traço essencial às diversas funções bioquímicas do organismo humano. As interações físico-químicas entre nutrientes incluem: adsorção, formação de complexos e precipitação, influenciando na biodisponibilidade. Através da determinação do teor total do metal ingerido, não é possível medir o quanto deste metal será absorvido. Entretanto, através da técnica de especiação dos metais, que é definida como a determinação da concentração das formas físico-químicas individuais dos elementos que, em conjunto, constitua a concentração total do mesmo na amostra, podemos ter novos dados para prever sua absorção. Para a especiação em amostras sólidas, inicialmente deve ser aplicado um método de extração seqüencial ou seletivo. Amostras de hortaliças dos tipos A e B foram analisadas quanto ao teor total de zinco através de espectroscopia de absorção atômica em chama, e o método de extração seqüencial foi aplicado com os seguintes extratores: solução de CaCl2 1M; solução de ácido acético 0,1M; solução de ácido acético 0,5M/acetato de amônio 5% [pH=5,0]; solução de NaOH 0,1M e solução de HCl 0,5M. Na análise da rejeição de resultados foi aplicado o teste de Grubbs. Em 8 amostras a extração do zinco foi considerada total, 6 amostras apresentaram extração superior a 75%, 5 amostras extração de 50% , amostra de quiabo teve 27% do total de zinco extraído e amostra de beterraba apenas 3%. Observa-se que para a maioria das amostras, o zinco se encontra sob a forma de no mínimo 6 espécies químicas distintas. Estudos posteriores, que permitam a identificação destas espécies, favorecerá a avaliação da biodisponibilidade deste metal.

Avaliação do uso de ervas medicinais como suplemento nutricional de ferro, cobre e zinco

O cobre, ferro e zinco, considerados elementos essenciais ao corpo humano, apresentam biodisponibilidade variável em função da forma química que se encontram em um alimento. As ervas medicinais, amplamente utilizadas, podem apresentar novas indicações quanto a suplementação destes metais. Este trabalho tem por objetivo avaliar os teores de cobre, ferro e zinco em ervas medicinais, pós e ervas secas, e promover a extração seqüencial visando a biodisponibilidade. Os teores de cobre, ferro e zinco foram determinados através da espectroscopia de absorção atômica. A extração seqüencial foi aplicada com os extratores cloreto de cálcio 1,0M; ácido acético 0,1M com acetato de amônio 5% (pH=5,0); ácido acético 0,5M e HCl 0,5M. Os resultados apresentaram teores altos de cobre, ferro e zinco, quando comparados com outras fontes alimentícias destes metais, além de indicar que os mesmos se apresentam sob, no mínimo, 4 espécies químicas distintas nas ervas analisadas. O extrator I foi o de melhor eficiência para os três metais. Considerando que o consumo destas ervas é feito com visão farmacológica, acredita-se que uso das mesmas em preparos de alimentos pode favorecer a suplementação dos metais cobre, ferro e zinco.

Conservação de maçã minimamente processada com o uso de películas comestíveis

O objetivo do trabalho foi avaliar o efeito de películas comestíveis na conservação de maçãs da cultivar Royal Gala Minimamente Processadas (MP) durante armazenamento refrigerado por 13 dias. Os tratamentos foram: o controle, a solução conservadora (1% ácido ascórbico; 0,5% ácido cítrico; 0,7% NaCl; 0,25% CaCl2) e as películas [dextrina (5,5%), fécula de mandioca (3%) e alginato de sódio (2%) com CaCl2 (0,6%)]. A solução conservadora reduziu a taxa respiratória e a produção de etileno em relação ao controle e promoveu maior claridade da polpa. Os tratamentos com películas apresentaram redução média de 38% na taxa respiratória e mais de 50% na produção de etileno em relação ao controle, sendo o alginato o mais eficiente. A película de alginato apresentou maior umidade no início do armazenamento, igualando-se aos demais tratamentos no 9º dia. No tratamento controle, os valores de firmeza dos tecidos diminuíram com o armazenamento e nos demais tratamentos, a tendência foi oposta. Maçãs com alginato apresentaram menor acidez e coloração mais escura dentre os tratamentos, respectivamente, em função do pH e da coloração da própria solução filmogênica.

Avaliação da eficiência de extração de compostos nitrogenados da polpa de anchoíta (Engraulis anchoita)

Este trabalho teve como objetivo avaliar a eficiência da extração de compostos nitrogenados, como proteínas solúveis (PS), nitrogênio total (NT), nitrogênio não proteico (NNP) e nitrogênio proteico (NP) da polpa de anchoíta (Engraulis anchoita), bem como obter informações a respeito de sua composição proximal e do frescor em relação ao seu local de captura. A polpa de anchoíta foi submetida a tratamento de extração de nitrogenados utilizando como soluções NaHCO3 (0,1; 0,2; 0,3 e 0,5%), NaCl 0,3% e água destilada. O ciclo de lavagem da polpa utilizando 0,1% de NaHCO3, dois ciclos de água destilada e um ciclo de NaCl 0,3% demonstrou maior eficiência na extração dos compostos nitrogenados, assim como das proteínas sarcoplasmáticas. Na determinação da composição proximal, a anchoíta in natura apresentou valores de umidade de 77,2%, proteína 16,8%, lipídios 3,4% e cinzas 2,4% e, para a polpa de anchoíta, foram encontrados valores de umidade de 78,1%, proteína 17,5%, lipídios 2,4% e cinzas 2,0%. A avaliação do frescor foi determinada através do pH, bases voláteis totais (N-BVT) e trimetilamina (N-TMA), encontrando-se valores de 6,3; 11,5 mg.100 g-1 e 2,8 mg.100 g-1 para a anchoíta in natura e 6,7; 20,2 mg.100 g-1; 3,1 mg.100 g-1 para a polpa, respectivamente.

Extraction, purification and characterization of inhibitor of trypsin from Chenopodium quinoa seeds

Abstract A novel trypsin inhibitor of protease (CqTI) was purified from Chenopodium quinoa seeds. The optimal extracting solvent was 0.1M NaCl pH 6.8 (p < 0.05). The extraction time of 5h and 90 °C was optimum for the recovery of the trypsin inhibitor from C. quinoa seeds. The purification occurred in gel-filtration and reverse phase chromatography. CqTI presented active against commercial bovine trypsin and chymotrypsin and had a specific activity of 5,033.00 (TIU/mg), which was purified to 333.5-fold. The extent of purification was determined by SDS-PAGE. CqTI had an apparent molecular weight of approximately 12KDa and two bands in reduced conditions as determined by Tricine-SDS-PAGE. MALDI-TOF showed two peaks in 4,246.5 and 7,908.18m/z. CqTI presented high levels of essential amino acids. N-terminal amino acid sequence of this protein did not show similarity to any known protease inhibitor. Its activity was stable over a pH range (2-12), temperatures range (20-100 °C) and reducing agents.