Kahweol, a natural diterpene from coffee, induces peripheral antinociception by endocannabinoid system activation

Kahweol is a compound derived from coffee with reported antinociceptive effects. Based on the few reports that exist in the literature regarding the mechanisms involved in kahweol-induced peripheral antinociceptive action, this study proposed to investigate the contribution of the endocannabinoid system to the peripheral antinociception induced in rats by kahweol. Hyperalgesia was induced by intraplantar injection of prostaglandin E2(PGE2) and was measured with the paw pressure test. Kahweol and the drugs to test the cannabinoid system were administered locally into the right hind paw. The endocannabinoids were purified by open-bed chromatography on silica and measured by LC-MS. Kahweol (80 µg/paw) induced peripheral antinociception against PGE2-induced hyperalgesia. This effect was reversed by the intraplantar injection of the CB1 cannabinoid receptor antagonist AM251 (20, 40, and 80 μg/paw), but not by the CB2 cannabinoid receptor antagonist AM630 (100 μg/paw). Treatment with the endocannabinoid reuptake inhibitor VDM11 (2.5 μg/paw) intensified the peripheral antinociceptive effect induced by low-dose kahweol (40 μg/paw). The monoacylglycerol lipase (MAGL) inhibitor, JZL184 (4 μg/paw), and the dual MAGL/fatty acid amide hydrolase (FAAH) inhibitor, MAFP (0.5 μg/paw), potentiated the peripheral antinociceptive effect of low-dose kahweol. Furthermore, kahweol increased the levels of the endocannabinoid anandamide, but not of the other endocannabinoid 2-arachidonoylglycerol nor of anandamide-related N-acylethanolamines, in the plantar surface of the rat paw. Our results suggested that kahweol induced peripheral antinociception via anandamide release and activation of CB1 cannabinoid receptors and this compound could be used to develop new drugs for pain relief.

Exposição crônica ao cloridrato de metformina e à glibenclamida causa alterações comportamentais, glicêmicas e de mortalidade em Hemigrammus caudovittatus e Danio rerio / Chronic exposure to metformin chloridrate and glibenclamide causes behavioral, blood glucose and mortality changes of Hemigrammus caudovittatus and Danio rerio

Hemigrammus caudovittatus e Danio rerio foram expostos aos hipoglicemiantes orais (HOs) cloridrato de metformina a 40µg/L e 120µg/L e glibenclamida a 0,13µg/L e 0,39µg/L durante 100 dias. Foram avaliados os efeitos tóxicos dos fármacos em relação ao peso, ao comportamento animal, à glicemia e à mortalidade. H. caudovittatus expostos à menor concentração dos fármacos apresentaram aumento significativo (P<0,05) no evento Respiração Aérea. Ainda, foi observado aumento no comportamento Descansar quando os animais foram expostos à glibenclamida a 0,39µg/L. Em D. rerio expostos ao cloridrato de metformina a 120µg/L, foi observado aumento (P<0,05) no comportamento Descansar. A glibenclamida provocou redução (P<0,05) na glicemia de H. caudovittatus. Ambos os fármacos causaram efeito letal na espécie D. rerio, contudo a glibenclamida foi mais tóxica, causando 100% de mortalidade em 30 dias de exposição. Os animais que vieram a óbito apresentaram congestão nos arcos branquiais e hemorragia. Os HOs foram desenvolvidos para apresentarem efeitos fisiológicos em mamíferos, entretanto efeitos tóxicos foram encontrados nas duas espécies de peixe estudadas. Isso levanta a preocupação sobre possíveis efeitos tóxicos de HOs e sobre quais métodos serão utilizados para a sua degradação no ambiente aquático.(AU)

Hemigrammus caudovittatus and Danio rerio were exposed to oral hypoglycemic drugs (HOs) metformin hydrochloride at 40µg/L and 120µg/L and to glibenclamide at 0.13µg/L and 0.39µg/L during 100 days. Toxic effects of the drugs were evaluated based on weight, animal behavior, blood glucose and mortality. H. caudovittatus exposed to lowest concentration of the drugs showed significant increase (P< 0.05) in the Air breathing event. Furthermore, increase in Rest event was observed when animals were exposed to glibenclamide at 0.39µg/L. An increase (P< 0.05) in the frequency of Rest behavior in the D. rerio exposed to metformin hydrochloride at 120µg/L was observed. Glibenclamide caused decrease (P< 0.05) in the blood glucose of H. caudovittatus. Both drugs caused lethal effect against D. rerio. Nevertheless, glibenclamide was more toxic causing 100% of mortality after 30 days of exposure. The animals that died showed congestion on the branchial arches and hemorrhage. The HOs were developed to have physiological effects in mammals. However, toxic effects were found in both species of fish studied. This raises concerns about possible toxic effects of HOs and what methods will be used for their degradation in the aquatic environment.(AU)

Avaliação hematológica e hemogasométrica de potros nascidos de éguas com placentite ascendente / Hematological and hemogasometric evaluation of foals born from mares with ascending placentitis

A placentite é a principal causa de partos prematuros, aborto e nascimento de potros comprometidos, podendo causar hipóxia e septicemia. A hematologia e a gasometria venosa fornecem informações importantes para o monitoramento da saúde de potros nascidos de éguas com placentite. O objetivo deste trabalho foi descrever os valores hematológicos e hemogasométricos durante as primeiras 24h de vida em potros nascidos de éguas mestiças Crioulas submetidas à indução experimental de placentite, com diferentes graus de maturidade. Foram utilizados 16 potros nascidos de éguas submetidas à indução experimental de placentite, divididos em três grupos de acordo com o grau de maturidade: prematuro (n=8), dismaturo (n=4) e a termo (n=4). Foram realizadas coletas sanguíneas nos momentos 0h, 12h e 24h para realização de hemograma completo e gasometria venosa. No eritrograma, foi observada anemia normocítica normocrômica no grupo prematuro em relação ao grupo dismaturo nas 12h e 24h. O grupo prematuro apresentou menor contagem de leucócitos totais nas 24h em relação ao grupo a termo (P=0,01). Os valores de pH, cHCO3 e SO2 não diferiram entre os grupos, porém os animais prematuros apresentaram acidose respiratória (pH=7,28). A PCO2 nos prematuros foi maior na 0h (P=0,02). Nos três grupos, a PCO2 apresentou uma curva adaptativa com redução dos valores durante as 12h e 24h. Os potros prematuros mostraram menores valores de excesso de base (BE) no nascimento (P=0,02), confirmando o quadro de acidose respiratória. Concluiu-se que as respostas hematológicas e hemogasométricas diferem entre potros com diferentes graus de maturidade. A acidose observada no grupo prematuro ao nascimento, com estabilização e resposta compensatória durante as 12h e 24h, demonstra a necessidade de avaliação hemogasométrica sequencial em potros de risco, o que permite a identificação da resposta clínica ao processo e, assim, auxilia no estabelecimento do tratamento e prognóstico para esses potros.(AU)

The placentitis is a major cause of premature birth, abortion and compromised foal delivery, and may result in hypoxia and sepsis. The blood gas analysis and hematology can provide important information for monitoring the foals born from mares with placentitis, with different degrees of maturity. The aim of this study was to describe the hematological and blood gas values during the first 24 hours of life in foals born from crossbreed mares with experimentally induced placentitis, presenting different degrees of maturity. Sixteen foals, born from mares with experimentally induced ascending placentitis were assigned to three groups according to degree of maturity: premature (n=8), dysmature (n=4), and full-term foals (n=4). Blood samples were collected at birth (0), at 12h and 24h, and hematological evaluation and blood gas variables were measured. In the premature group normocytic normochromic anemia was observed compared to dysmature group at 12h and 24h. The premature group showed lower count of white blood cells at 24h relative to the full-term group (p=0.01). The pH, cHCO3 and SO2 values do not differ among the groups; however the premature group showed respiratory acidosis (pH=7,28). The PCO2 was higher at 0h in the premature foals (p=0.02). In all groups, the PCO2 presented an adaptive curve with reduction between 12h and 24h. The premature foals showed lower base excess (BE) values at birth (p=0.02), confirming the respiratory acidosis in this group. We conclude that the hematological and blood gas response differs between foals with different degrees of maturity. Acidosis in the premature foals at birth, with stabilization and compensation of pH value during the first 12-24h demonstrate the necessity of sequential blood gas analysis in risk foals. This may help identify the clinical response to the process and assist in the establishment of adequate treatment and prognosis for these foals.(AU)

Regime alimentar para ganho compensatório de ovinos em confinamento: desempenho produtivo e morfometria do rúmen e do intestino delgado / Diet for compensatory gain of feedlot lambs: performance and morphometry of the rumen and small intestine

Objetivou-se avaliar o efeito do regime alimentar para ganho compensatório sobre o desempenho produtivo, o peso do corpo vazio, o peso dos componentes abióticos, o peso e a proporção do TGI e a morfometria do rúmen e do intestino delgado de cordeiros terminados em confinamento. Para tanto, foram utilizados 40 cordeiros Santa Inês, machos não castrados, alojados em baias individuais. O confinamento foi dividido em dois períodos distintos de 42 dias: um de restrição alimentar, com quatro tratamentos (0, 20, 40 e 60% de restrição), e outro de realimentação, em que todos os animais foram realimentados sem restrição. Ao final do período de confinamento, os cordeiros foram abatidos para mensuração do peso do TGI, do peso dos componentes abióticos e para obtenção dos fragmentos do rúmen e do intestino delgado, os quais foram posteriormente utilizados na confecção das lâminas histológicas usadas para avaliação da morfometria das papilas ruminais e das vilosidades e criptas do intestino (µm). As variáveis relativas ao desempenho, além de peso ao abate, peso do corpo vazio e peso do TGI, diminuíram à medida que se aumentou o nível de restrição prévia. A largura das papilas ruminais diminuiu, variando de 555,7 a 470,3µm; a área das vilosidades do intestino aumentou, variando de 81042,8 a 92033,7µm2. O regime alimentar para ganho compensatório diminuiu o ganho de peso, o peso ao abate, o peso do corpo vazio e o peso do trato gastrintestinal de cordeiros Santa Inês, terminados em confinamento, além de afetar a morfometria das papilas ruminais e das vilosidades do intestino delgado...

The aim was evaluate the effect of diet for compensatory growth on performance, empty weight body, weight of abiotic components, weight and proportion of TGI and morphology of the rumen and small intestine of lambs feedlot. Thus, we used 40 Santa Inês non castrated male lambs, housed in individual pens. The containment was divided into two distinct periods of 42 days: a food restriction, with four treatments (0, 20, 40 and 60% restriction), and a feedback loop, where all the animals were fed again without restriction. At the end of the period of confinement they were slaughtered to measure the weight of TGI, weight and abiotic components for obtaining fragments of the rumen and small intestine, which were subsequently used for the preparation of histological slides used to evaluate the morphology of the papillae rumen and the intestinal villi and crypts (μm). The variables for performance, slaughter weight, empty body weight and weight of TGI decreased as they increased the level of prior restraint. The width of the rumen papillae decreased ranging from 555.7 to 470.3μm; the area of increased intestinal villi ranged from 81042.8 to 92033.7μm2. The diet for compensatory gain decreases weight gain, slaughter weight, empty body weight and the weight of the gastrointestinal tract of Santa Ines lambs, besides affecting the morphology of rumen papillae and villi of the small intestine...

Substituição de milho por glicerina bruta em dietas para suínos em terminação / Substitution of corn for crude glycerin in diets for finishing pigs

Avaliou-se o efeito da substituição do milho por glicerina bruta em dietas para suínos em terminação. Foram utilizados 80 suínos, machos castrados, híbrido comercial, com média de peso de 67kg, em um experimento em delineamento inteiramente ao acaso, com cinco tratamentos e oito repetições, com dois animais por unidade experimental. Os tratamentos foram: 0,0; 4,0; 8,0; 12,0 e 16,0% de glicerina bruta em substituição ao milho nas dietas. A substituição do milho por glicerina bruta não afetou as características de desempenho (P>0,05). Observaram-se efeito linear crescente de tratamento (P<0,05) sobre a espessura de toucinho na carcaça e efeito linear decrescente de tratamento (P<0,05) sobre a perda de líquido no descongelamento e na força de cisalhamento. A glicerina bruta pode substituir em até 16,0% do milho da dieta para suínos em terminação, sem prejuízos do desempenho e rendimento de carcaça, com melhoras na qualidade da carne.

We evaluated the effect of replacing corn with crude glycerin in diets for finishing pigs. We used 80 pigs, steers, commercial hybrids, with an average weight of 67kg, in an experiment with a completely randomized design with five treatments and eight replicates of two animals per experimental unit. The treatments were 0.0, 4.0, 8.0, 12.0, and 16.0% crude glycerin as a replacement for maize in diets. The substitution of corn with crude glycerin did not affect performance characteristics (P>0.05). We observed an increasing linear effect of the treatment (P<0.05) on backfat thickness and decreasing linear effect of the treatment (P<0.05) on the loss of fluid in the thawing and shearing force. The crude glycerin can replace up to 16.0% of the corn diet for finishing pigs without loss of performance and carcass yield, with improvements in meat quality.

Antibody responses elicited in mice immunized with Bacillus subtilis vaccine strains expressing Stx2B subunit of enterohaemorragic Escherichia coli O157: H7 / Resposta de anticorpos obtidas em camundongos imunizados com linhagens vacinais de Bacillus subtilis expressando a subunidade B da Stx2 de Escherichia coli O157: H7 enterohemorrágica

No effective vaccine or immunotherapy is presently available for patients with the hemolytic uremic syndrome (HUS) induced by Shiga-like toxin (Stx) producedbyenterohaemorragic Escherichia coli (EHEC) strains, such as those belonging to the O157:H7 serotype. In this work we evaluated the performance of Bacillus subtilis strains, a harmless spore former gram-positive bacterium species, as a vaccine vehicle for the expression of Stx2B subunit (Stx2B). A recombinant B. subtilis vaccine strain expressing Stx2B under the control of a stress inducible promoter was delivered to BALB/c mice via oral, nasal or subcutaneous routes using both vegetative cells and spores. Mice immunized with vegetative cells by the oral route developed low but specific anti-Stx2B serum IgG and fecal IgA responses while mice immunized with recombinant spores developed anti-Stx2B responses only after administration via the parenteral route. Nonetheless, serum anti-Stx2B antibodies raised in mice immunized with the recombinant B. subtilis strain did not inhibit the toxic effects of the native toxin, both under in vitro and in vivo conditions, suggesting that either the quantity or the quality of the induced immune response did not support an effective neutralization of Stx2 produced by EHEC strains.

Até o presente o momento, não há vacina ou imunoterapia disponível para pacientes com Síndrome Hemolítica Urêmica (SHU) induzida pela toxina Shiga-like (Stx) produzida por linhagens de Escherichia coli entero-hemorragica (EHEC), tais como as pertencentes ao sorotipo O157:H7. Neste trabalho, avaliamos a performance de Bacillus subtilis, uma espécie bacteriana gram-positiva não-patogênica formadora de esporos, como veículo vacinal para a expressão da subunidade B da Stx2B (Stx2B). Uma linhagem vacinal recombinante de B. subtilis expressando Stx2B, sob o controle de um promoter induzível por estresse, foi administrada a camundongos BALB/c por via oral, nasal ou subcutânea usando células vegetativas e esporos. Camundongos imunizados com células vegetativas e esporos pela via oral desenvolveram títulos anti-Stx2B baixos, mas específicos, de IgG sérico e IgA fecal, enquanto camundongos imunizados com esporos recombinates desenvolveram resposta anti-Stx2B apenas após a administração pela via parenteral. No entanto, anticorpos produzidos em camundongos imunizados com a linhagem recombinante de B. subtilis não inibiram os efeitos tóxicos da toxina nativa em condições in vitro e in vivo, sugerindo que a quantidade e/ou a qualidade da resposta imune gerada não suportam uma neutralização efetiva da Stx2 produzidas por linhagens de EHEC.

Production of the refolded oligopeptide-binding protein (OppA) encoded by the citrus pathogen Xanthomonas axonopodis pv. citri

The oligopeptide-binding protein, OppA, binds and ushers oligopeptide substrates to the membrane-associated oligopeptide permease (Opp), a multi-component ABC-type transporter involved in the uptake of oligopeptides expressed by several bacterial species. In the present study, we report the cloning, purification, refolding and conformational analysis of a recombinant OppA protein derived from Xanthomonas axonopodis pv. citri (X. citri), the etiological agent of citrus canker. The oppA gene was expressed in Escherichia coli BL21 (DE3) strain under optimized inducing conditions and the recombinant protein remained largely insoluble. Solubilization was achieved following refolding of the denatured protein. Circular dichroism analysis indicated that the recombinant OppA protein preserved conformational features of orthologs expressed by other bacterial species. The refolded recombinant OppA represents a useful tool for structural and functional analyses of the X. citri protein.

Structural model and ligand interactions of the Xanthomonas axonopodis pv. citri oligopeptide-binding protein

The oligopeptide-binding protein, OppA, ushers oligopeptide substrates to the membrane-associated oligopeptide permease (Opp), a multi-component ABC-type transporter involved in the uptake of oligopeptides by several bacterial species. In the present study, we report a structural model and an oligopeptide docking analysis of the OppA protein expressed by Xanthomonas axonopodis pv. citri (X. citri), the etiological agent of citrus canker. The X. citri OppA structural model showed a conserved three-dimensional structure, irrespective of the low amino acid identities with previously defined structures of Bacillus subtilis and Salmonella typhimurium orthologs. Oligopeptide docking analysis carried out with the proposed model indicated that the X. citri OppA preferentially binds tri- and tetrapeptides. The present study represents the first structural analysis of an OppA ortholog expressed by a phytopathogen and contributes to the understanding of the physiology and nutritional strategies of X. citri.

Minimization of transcriptional temporal noise and scale invariance in the yeast genome

The analysis of transcriptional temporal noise could be an interesting means to study gene expression dynamics and stochasticity in eukaryotes. To study the statistical distributions of temporal noise in the eukaryotic model system Saccharomyces cerevisiae, we analyzed microarray data corresponding to one cell cycle for 6200 genes. We found that the temporal noise follows a lognormal distribution with scale invariance at the genome, chromosomal and sub-chromosomal levels. Correlation of temporal noise with the codon adaptation index suggests that at least 70% of all protein-coding genes are a noise minimization core of the genome. Accordingly, a mathematical model of individual gene expression dynamics was proposed, using an operator theoretical approach, which reveals strict conditions for noise variability and a possible global noise minimization/optimization strategy at the genome level. Our model and data show that minimal noise does not correspond to genes obeying a strictly deterministic dynamics. The natural strategy of minimization consists in equating the mean of the absolute value of the relative variation of the expression level (alpha) with noise (eta). We hypothesize that the temporal noise pattern is an emergent property of the genome and shows how the dynamics of gene expression could be related to chromosomal organization.

Diabetes mellitus: new therapeutic approaches to treat an old disease

Diabetes mellitus is a widespread disease whose frequency increases constantly and is expected to reach alarming levels by the year 2025. Introduction of insulin therapy represented a major breakthrough; however, a very strict regimen is required to maintain blood glucose levels within the normal range and to prevent or postpone chronic complications associated with this disease. Frequent hyper- and hypoglycemia seriously affect the quality of life of these patients. Reversion of this situation can only be achieved through whole organ (pancreas) transplant or pancreatic islet transplant, the former being a high-risk surgical procedure, while the latter is a much simpler and may be accomplished in only 20-40 min. The advantages and perspectives of islet cell transplantation will be discussed, in the light of tissue engineering and gene therapy. Ongoing research carried out in our laboratory, aimed at developing clinical cell and molecular therapy protocols for diabetes will also be focused

Environmental and genetic factors affecting mutability to aminoglycoside antibiotics among Escherichia coli K12 strains

Environmental and genetic factors affecting the in vitro spontaneous mutation frequencies to aminoglycoside resistance in Escherichia coli K12 were investigated. Spontaneous mutation frequencies to kanamycin resistance were at least 100 fold higher on modified Luria agar (L2) plates, when compared to results obtained in experiments carried out with Nutrient agar (NA) plates. In contrast to rifampincin, the increased mutability to kanamycin resistance could not be attributed to a mutator phenotype expressed by DNA repair defective strains. Kanamycin mutant selection windows and mutant preventive concentrations on L2 plates were at least fourfold higher than on NA plates, further demonstrating the role of growth medium composition on the mutability to aminoglycosides. Mutability to kanamycin resistance was increased following addition of sorbitol, suggesting that osmolarity is involved on the spontaneous mutability of E. coli K12 strains to aminoglycosides. The spontaneous mutation rates to kanamycin resistance on both L2 and NA plates were strictly associated with the selective antibiotic concentrations. Moreover, mutants selected at different antibiotic concentrations expressed heterogeneous resistance levels to kanamycin and most of them expressing multiple resistance to all tested aminoglycoside antibiotics (gentamicin, neomycin, amykacin and tobramycin). These results will contribute to a better understanding of the complex nature of aminoglycoside resistance and the emergence of spontaneous resistant mutants among E. coli K12 strains

Efficacy and safety of risperidone oral solution in agitation associated with dementia in the elderly

BACKGROUND: Behavioral and psychological symptoms in dementia (BPSD) contribute to caregiver burden and institutionalization of elderly. Neuroleptics are prescribed to control agitation. Side effects of typical neuroleptics are harmful, making atypical neuroleptics an indication. OBJECTIVES: To evaluate efficacy and tolerability of risperidone oral solution (ROS) given once daily to demented elderly outpatients with BPSD (agitation). METHOD: Patients (n=26), 76.35 + or 8.63 years, Diagnostic and Statistical Manual of Mental Disorders 4th ed. (DSM-IV) criteria for dementia. RSO was given, starting dose of 0.25 mg and increments of 0.25 mg every week. Mini-Mental State Examination (MMSE) assessed cognitive status, Behavioral and Emotional Activities Manifested in Dementia (BEAM-D) and Clinical Global Impression (CGI) measured BPSD, Extrapiramidal Symptom Rating Scale (ESRS) evaluated extrapyramidal symptoms. Cardiovascular side effects were evaluated clinically. RESULTS: There was a 26 percent reduction in agitation and no cardiovascular side effects in the range from 1.0 to 1.25 mg. Side effects were more prevalent above 2.5 mg. CONCLUSION: Risperidone oral solution improved agitation with good tolerability from 0.5 to 1.25 mg. A single dose with increments of 0.25 mg may be more acceptable to patients and caregivers

Evaluation of genotoxic activity in the blood and urine of guinea pigs treated with nifurtimox and benznidazole

1. The mutagenicity of serum and urine from fuinea pigs treated with a single oral dose (500 mg/Kg) of benznidazole and nifurtimox was assayed using the Salmonella/plate incorporation test with strain TA100 and a nitroreductase-deficient derivative, TA100NR. 2. The urine and blood of animals treated with nifurtimox were not mutagenic for either tester strain. 3. The urine and blood of animals receiving benznidazole were mutagenic to the TA100 but not to the TA100NR strain. Similar results were obtained with nitrofurantoin-treated animals. Maximum mutagenicity values were obtained in serum and urine of treated animals 90 min and 24 h after administration, respectively. 4. Mutagenicity induced by benznidazole in the serum and urine of treated animals was not altered when assayed in anaerobic environments. 5. These results indicate that benznidazole and nifurtimox are not metabolized by the mammalian host into stable mutagenic derivatives detectable by the Ames test. Based on these data, we suggest that the potential cancer risk to patients treated with these drugs is small but should be further evaluated

CL 64, 855, a potent do anti-Trypanosoma cruzi drug, is also mutagenic in the salmonella/microsome assay

O derivado nitroimizadole-tiadizol CL 64.855 (2-amino-5-(1-metil-5-nitro-2-imidazoli)-1, 3, 4-tiadiazol), um potente agente tripanomicida, foi submetido a um ensaio mutagênico bacteriano com as linhagens indicadoras de Salmonella typhimurium TA 98, TA 100 e TA 102. Os resultados indicaram que o CL 64.855 é um potente mutagênico tipo troca de referencial detectado pelas linhagens TA 98 e TA 102. O CL 64.855 foi capaz de reverter as linhagens indicadoras em concentraçöes täo baixas quatro 0,1microng/placa. Ativaçäo metabólica com fraçöes microssomais de fígado de rato foram incapazes de aumentar a açäo mutagênica do CL 64.855