RESUMO Hyptis pectinata, conhecida popularmente como Sambacaitá é uma erva medicinal nativa utilizada pela população local para o tratamento de enfermidades, cujas propriedades farmacológicas, antimicrobianas e medicinais foram relatadas em diversos estudos. O óleo essencial das folhas é rico em compostos terpênicos responsáveis pelas suas propriedades. Devido à existência de fatores que podem influenciar a composição química dos óleos essenciais, o objetivo deste trabalho foi avaliar a influência do armazenamento do óleo essencial Hyptis pectinata durante 360 dias em duas temperaturas diferentes. O óleo essencial foi destilado a partir de folhas secas de plantas de sete meses de idade, utilizando o método de hidrodestilação e a composição química foi determinada usando CG-EM e CG-DIC. Os efeitos do armazenamento no óleo essencial Hyptis pectinata foram testados ao longo de um ano (0, 15, 30, 60, 90, 120, 150, 180, 240, 300 e 360 dias), utilizando duas temperaturas: ambiente (±32ºC) e freezer (-20ºC). Os compostos predominantes no óleo essencial foram o β-cariofileno, óxido de cariofileno e germancreno D e houve variação na concentração dos constituintes químicos do óleo essencial, dependendo da temperatura de armazenamento, bem como ao longo dos 360 dias. O armazenamento do óleo essencial, à temperatura ambiente ao longo de um ano resultou em maiores concentrações de β-elemeno, α-copaeno, germacreno D, óxido de cariofileno e (E, E) -α-farneseno e menores teores de α-humuleno e β-cariofileno, quando comparado com os resultados obtidos a partir de armazenamento do óleo essencial no freezer. Estes resultados indicam que o óleo essencial deve ser estocado no freezer, o que propicia maior estabilidade na concentração dos constituintes químicos.

ABSTRACT Hyptis pectinata, popularly known as “sambacaitá”, is a native medicinal herb used by local people for treating disease. Its pharmacological, antimicrobial, and medicinal properties have been reported in several studies. The essential oil of the leaves is rich in terpene compounds responsible for its properties. Because there are factors that can influence the chemical composition of essential oils, the objective of this study was to evaluate the influence of storage on Hyptis pectinata essential oil for 360 days at two different temperatures. The essential oil was distilled from dried leaves of seven-month-old plants using the hydrodistillation method, and the chemical composition was determined using GC-MS and GC/FID. The effects of storage on the essential oil Hyptis pectinata were tested over the course of a year (0, 15, 30, 60, 90, 120, 150, 180, 240, 300, and 360 days), using two temperatures: room (±32ºC) and freezer (-20ºC). The predominant compounds in the essential oil are β–caryophyllene, caryophyllene oxide, and germancrene D. The concentration of the chemical constituents of the essential oil varied depending on the storage temperature, and over the 360 days. The storage of the essential oil at room temperature over one year resulted in higher concentrations of β-elemene, α-copaene, germacrene D, caryophyllene oxide, and (E,E)-α-farnesene and lower content of α-humulene and β-caryophyllene, compared to the results obtained from storing the essential oil in a freezer. These results indicate that the essential oil should be stored in a freezer, which provides greater stability to the concentration of the chemical constituents.

Sex hormones modulate the action of both cytokines and the renin-angiotensin system. However, the effects of angiotensin I-converting enzyme (ACE) on the proinflammatory and anti-inflammatory cytokine levels in male and female spontaneously hypertensive rats (SHR) are unclear. We determined the relationship between ACE activity, cytokine levels and sex differences in SHR. Female (F) and male (M) SHR were divided into 4 experimental groups each (n = 7): sham + vehicle (SV), sham + enalapril (10 mg/kg body weight by gavage), castrated + vehicle, and castrated + enalapril. Treatment began 21 days after castration and continued for 30 days. Serum cytokine levels (ELISA) and ACE activity (fluorimetry) were measured. Male rats exhibited a higher serum ACE activity than female rats. Castration reduced serum ACE in males but did not affect it in females. Enalapril reduced serum ACE in all groups. IL-10 (FSV = 16.4 ± 1.1 pg/mL; MSV = 12.8 ± 1.2 pg/mL), TNF-α (FSV = 16.6 ± 1.2 pg/mL; MSV = 12.8 ± 1 pg/mL) and IL-6 (FSV = 10.3 ± 0.2 pg/mL; MSV = 7.2 ± 0.2 pg/mL) levels were higher in females than in males. Ovariectomy reduced all cytokine levels and orchiectomy reduced IL-6 but increased IL-10 concentrations in males. Castration eliminated the differences in all inflammatory cytokine levels (IL-6 and TNF-α) between males and females. Enalapril increased IL-10 in all groups and reduced IL-6 in SV rats. In conclusion, serum ACE inhibition by enalapril eliminated the sexual dimorphisms of cytokine levels in SV animals, which suggests that enalapril exerts systemic anti-inflammatory and anti-hypertensive effects.

O Mycoplasma é considerado cosmopolita, podendo ser disseminado através do comércio internacional de animais, sêmen industrializado e de produtos de transferência de embriões. A expansão de células do cumulus é utilizada como parâmetro de avaliação de oócitos bovinos cultivados in vitro e suas alterações morfológicas são representativas. O objetivo deste trabalho foi avaliar a interação do Mycoplasma bovigenitalium, exposto experimentalmente à cultura primária de célula do cumulus, após o período de maturação. Complexos oócitos cumulus (COCs) obtidos através de punção folicular de ovários bovinos, provenientes de abatedouro, foram divididos em dois grupos para serem maturados durante 24h em meio de maturação (TCM199 + hormônios) em estufa a 38º C, 5% de CO2, 95% de umidade. Posteriormente, os oócitos foram retirados das placas, permanecendo somente com as células do cumulus aderidas. Com o monoestrato celular formado, um grupo foi infectado com 30 mL de M. bovigenitalium, replicado em meio Hayflick modificado a 37º C em estufa de microaerofilia, enquanto o outro foi mantido como controle. Os resultados mostraram que, com 24h de exposição ao patógeno, as culturas apresentaram um pequeno número de células arredondadas e granulosas, quando comparadas as dos controles. Esse efeito persistiu até o sétimo dia, onde se iniciou um processo de descolamento das células. Pode-se concluir que uma contaminação por micoplasma pode ser imperceptível às manipulações da FIV, pois células infectadas por esse grupo de bactérias não apresentam turvações no meio de cultura e, quando não lisam a célula hospedeira, tornam mais suscetível ao ambiente e outros agentes infecciosos.

The Mycoplasma is considered cosmopolitan and can be disseminated through international trade of animals, industrialized semen and embryo transfer products. The expansion of cumulus cells is used as a parameter for evaluating cattle oocytes cultivated in vitro, and their morphological changes are representative. The aim of the present study was to evaluate the interaction of Mycoplasma bovigenitalium with primary-culture cumulus cells, after the in vitro maturation period. Cumulus complex oocytes (COCs) obtained through follicular puncture of ovaries from a cattle slaughterhouse were divided into two groups to be matured for 24 hours in the maturation medium (TCM 199 + hormones) in a climate controlled chamber at 38º C, 5% CO2, and 95% humidity. Subsequently, the oocytes were removed from the plates, which remained with only the cumulus cells attached. With the monostratum cell formed, one group was infected with 30 ºL (5 x 10(6) cells/mL) M. bovigenitalium, replicated in modified Hayflick medium at 37º C in a mycroaerofilic chamber, while the other was kept as a control. The results showed that with 24 hours of exposure to the pathogen, the cultures showed a small number of rounded and grainy cells, when compared to the controls. This effect persisted until the 7th day, when a process of cell detachment began. It can be concluded that a mycoplasma contamination may be imperceptible to the manipulations of FIV, because cells infected by this group of bacteria present no cloudiness in the culture field, and when they do not lyse the host cell, they make it more susceptible to environment and other infectious agents.

A continuous assay using internally quenched fluorescent peptides with the general sequence Abz-peptidyl-(Dnp)P-OH (Abz = ortho-aminobenzoic acid; Dnp = 2,4-dinitrophenyl) was optimized for the measurement of angiotensin I-converting enzyme (ACE) in human plasma and rat tissues. Abz-FRK(Dnp)P-OH, which was cleaved at the Arg-Lys bond by ACE, was used for the enzyme evaluation in human plasma. Enzymatic activity was monitored by continuous recording of the fluorescence (lambdaex = 320 nm and lambdaem = 420 nm) at 37ºC, in 0.1 M Tris-HCl buffer, pH 7.0, with 50 mM NaCl and 10 µM ZnCl2. The assays can be performed directly in the cuvette of the fluorimeter and the hydrolysis followed for 5 to 10 min. ACE measurements in the plasma of 80 healthy patients with Hip-His-Leu and with Abz-FRK(Dnp)P-OH correlated closely (r = 0.90, P < 0.001). The specificity of the assay was demonstrated by the complete inhibition of hydrolysis by 0.5 µM lisinopril or captopril. Abz-FRK(Dnp)P-OH cleavage by ACE was monitored in rat lung, kidney, heart, and liver homogenates in the presence of a cocktail of inhibitors containing trans-epoxy-succinyl-L-leucylamido-(4-guanido)-butene, pepstatin, phenyl-methylsulfonyl fluoride, N-tosyl-L-phenylalanyl-chloromethyl ketone, and N-tosyl-lysyl-chloromethyl ketone to prevent undesirable hydrolysis. ACE activity in lung, heart and kidney homogenates, but not in liver homogenates, was completely abolished by 0.5 µM lisinopril or captopril. The advantages of the method are the procedural simplicity and the high sensitivity providing a rapid assay for ACE determinations.