[Antimicrobial susceptibility of no-O1 Vibrio cholerae strains isolated from patients in Cuba].
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Biomedical subjects
Publications and source records attributed to M Ramírez.
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Previous studies have demonstrated that FSH stimulates cell proliferation in the ovary and the testis of the chick embryo. This study analyzed the presence of FSH receptor and the cell subpopulations that proliferate in response to FSH in chick embryo gonads. FSH receptor mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) in the male and female gonads of the 6 to 14-day-old chick embryo. Somatic cells of the ovary expressed the FSH receptor in the 14-day-old chick embryo. Ovarian surface epithelium of the 14-day-old chick embryo increased the mitotic index 15-18 h after FSH treatment. Similarly, the mitotic index in oogonia was increased 24 h after receiving a pulse of FSH; this result was confirmed by an increase in the number of germ cells that incorporated bromodeoxyuridine (BrdU). Somatic cells of the medullary cords in the ovary displayed an increase in the mitotic index 15-21 h after the FSH injection. In the chick embryo testis, at the same stage of development, the treatment with FSH increased the mitotic index in cells of the seminiferous tubules and to a lesser extent in cells at a peritubular and interstitial location. Present results demonstrate that in the chick embryo, FSH stimulates the proliferation of ovarian surface epithelium, oogonia in the cortex, and somatic cells of the medullary cords of the chick embryo ovary. In the chick embryo testis, FSH stimulates cell proliferation in seminiferous tubules and peritubular cells.
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A local renin-angiotensin system (RAS) that may be involved in their regulatory functions has been identified in hypothalamus and pituitary. Altered thyroid status induces modifications in the secretory function of hypothalamus and pituitary. However, few studies have analyzed the role of the RAS in hypothalamus and, to our knowledge, there is no data on the pituitary RAS during thyroid dysfunction. In the present study, angiotensinase activities (glutamyl, aspartyl and alanyl aminopeptidase: GluAP, AspAP and AlaAP, respectively) were studied in hypothalamus and in the anterior and posterior lobes of pituitary of euthyroid, hypothyroid and hyperthyroid adult male rats. In the anterior pituitary, compared with euthyroid and hyperthyroid rats, hypothyroid animals showed a highly significant increase of GluAP and AspAP activities; the percentage increase in GluAP was markedly higher than the percentage increase in AspAP. This suggests an increased metabolism of angiotensin (Ang) I and Ang II to des-Asp 1-Ang I and Ang III, respectively. We also observed an increase of Ang III-degrading activity (AlaAP) in the hypothalamus of hyperthyroid rats in soluble fraction. Increased Ang I and Ang II metabolism in the anterior pituitary of hypothyroid rats and increased metabolism of Ang III in the hypothalamus of hyperthyroid animals may be related to alterations in the secretory function of hypothalamus and pituitary in these thyroid dysfunctions.
We compared the Etest with a broth microdilution method, performed according to a modified National Committee for Clinical Laboratory Standards guideline (M38-A), for determining the in vitro susceptibility of 77 isolates of Aspergillus spp. (26 A. fumigatus, 21 A. flavus, 10 A. terreus, 9 A. niger, 5 A. nidulellus, 4 A. glaucus, and 2 A. flavipes isolates). Overall, there was 92.2% agreement between both methods when Etest MICs were read at 24 h and 83.1% agreement when both methods were read at 48 h. When Etest MICs were read at 24 h, the agreement was >90% for all species tested except for A. fumigatus (84.6%). When Etest MICs were read at 48 h, the agreement ranged from 50 to 100%. The poorest agreement was seen with A. glaucus (50%) and A. fumigatus (65%). Where a discrepancy was observed between Etest and the reference method, the Etest MIC was generally higher. The Etest appears to be a suitable alternative procedure for testing the susceptibility of Aspergillus spp. to voriconazole.
Several studies have addressed the interaction between fatty acids and lipids with central nervous system peptides. Because aminopeptidases (AP) are involved in the regulation of neuropeptides, this work studies several AP expressed in cultured astroglia, after exogenous addition of oleic and linoleic fatty acids and cholesterol to the culture medium. Alanyl-AP, arginyl-AP, cystyl-AP, leucyl-AP, tyrosyl-AP and pyroglutamyl-AP activities were analysed in whole cells using the corresponding aminoacyl-beta-naphthylamides as substrates. Oleic acid inhibits alanyl-AP, cystyl-AP and leucyl-AP activities, whereas linoleic acid inhibits alanyl-AP, arginyl-AP and tyrosyl-AP activities. Neither oleic acid nor linoleic acid modifies pyroglutamyl-AP activity. In contrast, cholesterol increases arginyl-AP, cystyl-AP, leucyl-AP, tyrosyl-AP and pyroglutamyl-AP activities, although it does not modify alanyl-AP activity. The changes reported here suggest that oleic and linoleic fatty acids and cholesterol can modulate peptide activities via their degradation route involving aminopeptidases; each of them being differentially regulated.
Although the granule cells of the dentate gyrus are glutamatergic, they contain the machinery for the synthesis and vesiculation of GABA. Furthermore, glutamic acid decarboxylase and the vesicular GABA transporter mRNA are expressed in the granule cells and mossy fibers in an activity-dependent manner, suggesting that these cells release GABA in addition to glutamate. Supporting this hypothesis, we found that seizures induce simultaneous glutamatergic and GABAergic transmission in the mossy fiber projection. To further explore this expression of inhibition, we looked for the presence and expression of endogenous GABA in a synaptosomal preparation enriched with mossy fiber nerve endings of kindled rats. We also studied the capacity of this preparation to capture and release [(3)H]GABA under control conditions and after kindling epilepsy. In accordance with our hypothesis we show that the mossy fiber synaptosomal preparation of the kindled rats has a significantly higher content of endogenous GABA than controls. We also found that the protein content in the mossy fiber synaptosomal preparation of kindled rats was significantly augmented, which is consistent with mossy fiber sprouting. Due to this, the total [(3)H]GABA incorporated in the synaptosomal preparation was also augmented. However, [(3)H]GABA uptake (expressed in % radioactivity/mg protein) and its evoked release were similar in both groups. With the present results, we provide further support for the hypothesis of the emergence of GABAergic transmission in the mossy fiber synapse that can constitute a protective mechanism in response to seizures.
The presence of mononuclear cells infiltrating the prostate adenoma is a morphological observation well established in the literature. However, its biological meaning is a subject of controversy. It has been postulated that it may represent a local immunological reaction contributing to the pathogenesis of prostatic adenoma. Several studies have been performed to test this hypothesis, both in humans and animals. The purpose of this review is to update available information, including our own ongoing studies. Morphological research has shown that cells infiltrating the adenoma are lymphocyte T, lymphocyte B and macrophages with a high proportion of lymphocyte T. Many of the inflammatory markers, such as lymphoquines (IL1, IL2, IL4, IL6, IL13), are elevated in the adenoma tissue as are some growth factors (EGF, TGF alpha, IFN gamma, TGF beta). The general impression is that an inflammatory process is activated in the adenoma during growth and maturing. It has also been proved that this inflammatory process could be modified with treatment and, in our case, with the lipido-sterolic extract of Serenoa Repens.
Given that the existence of a local renin-angiotensin system (RAS) in the pituitary and its participation in the regulation of blood pressure and other biological functions are widely accepted, the aim of this work is to analyze the influence of dietary cholesterol on the activity of the enzymes involved in the metabolism of the effector peptides of the renin-angiotensin system (angiotensin II and III) and vasopressin, in the pituitary of male and female mice fed on a cholesterol-enriched diet (1% cholesterol and 0.5% cholic acid). Soluble and membrane-bound pituitary aminopeptidase A (aspartyl- and glutamyl-aminopeptidase), aminopeptidase M (alanyl-aminopeptidase), aminopeptidase B (arginyl-aminopeptidase) and cystinyl-aminopeptidase activities were fluorimetrically measured. In female mice, cholesterol-enriched diet produced a significant increase in soluble aspartyl- and membrane-bound aspartyl- and glutamyl-aminopeptidase activities, and a significant decrease in membrane-bound alanyl-, arginyl- and cystinyl-aminopeptidase activities. In male mice, after feeding the diet, a significant increase in soluble glutamyl- and membrane-bound arginyl-aminopeptidase activities was observed. Our results indicate differential effects of dietary cholesterol on the metabolism of angiotensin II and III and vasopressin in the pituitary of male and female mice.
In the normal granule cells of the dentate gyrus glutamate, GABA and glutamic acid decarboxylase (GAD67) coexist. After kindled seizures, this enzyme is transiently overexpressed and simultaneous glutamatergic and GABAergic transmission in the mossy fiber projection occurs. Since this dual transmission is also seen after acutely-induced seizures, we decided to study the relationship between the expression of GAD67 and the induction of simultaneous glutamatergic and GABAergic transmission by kindled or acutely induced seizures. We also explored whether kindling of the dentate gyrus in vitro, that does not induce epileptiform activity, could induce the expression of GAD67. We confirm that kindling epilepsy induces the expression of GAD67 in the dentate gyrus. Despite the emergence of GABAergic transmission in the mossy fiber projection after a single seizure, GAD67 expression in the dentate gyrus appeared similar to controls, however, in the mossy fibers an enhanced immunostaining was evident. Interestingly, kindling the dentate gyrus in vitro induces a marked GAD67 staining in the granule cells. Our results show that after the activity-dependent emergence of simultaneous glutamatergic and GABAergic transmission from the mossy fibers, GAD67 is expressed in the mossy fibers and, upon long-lasting enduring stimulation periods, also in the dentate gyrus. Thus, this phenomenon does not depend on the presence of epileptic activity, but rather, on increased excitatory input onto the dentate gyrus. This can represent a protective mechanism that can sustain GABA synthesis in an activity-dependent manner.
Aminopeptidases (APs) play a major role in the metabolism of circulating and local peptides, such as angiotensins and vasopressin, substances involved in the control of blood pressure and water balance. In the present work, we studied the influence of dehydration on angiotensinases and vasopressin-degrading activity. Since sex differences may exist in the regulation of water balance by angiotensin II and differential sexual steroid modulation of vasopressin secretion, in response to osmotic stimulation have been reported, gonadotropin releasing hormone (GnRH)-degrading activity was also analysed in serum, neurohypophysis and adrenal glands of male and female rats. Our results did not suggest sex differences in the response to changes in osmolality. GnRH-degrading activity decreased in serum of dehydrated males and females, which suggests a longer action of the peptide under these conditions. In neurohypophysis, there was an increase in the activity of aminopeptidase A (APA), the enzyme responsible for the metabolism of angiotensin II to angiotensin III. This occurs with a decrease in alanyl aminopeptidase activity, which would lead to a prolonged action of angiotensin III by reduction of its metabolism. In adrenals of dehydrated animals, the results would imply a high degree of metabolism of angiotensin III and vasopressin.
We evaluated the influence of a diet supplemented with olive oil (20% by weight) (OO) on the activity of glutamyl aminopeptidase (GluAP) and aspartyl aminopeptidase (AspAP), which are involved in angiotensin metabolism. Serum concentrations of total cholesterol and fatty acids were also measured. Animals fed on the OO diet gained significantly more weight than did controls from the second week until the end of the feeding period. Serum total cholesterol concentration was significantly higher in the OO group than in control mice. Total monounsaturated fatty acids increased in OO-fed animals, but total saturated fatty acids decreased. No differences between the two groups were observed for total polyunsaturated fatty acids. Serum from animals fed on the OO diet contained significantly lower proportions of myristic, pentadecanoic, palmitic, palmitoleic, vaccenic, alpha-linolenic, gamma-linolenic, and 11,14-eicosadienoic acids than did serum from control animals. In contrast, the OO group had higher levels of oleic, stearic, and gondoic acids. GluAP activity decreased significantly in the serum of OO-fed animals. In these animals soluble AspAP activity was significantly higher in the testis, and significantly lower in the lung and adrenal, in comparison to controls. Membrane-bound AspAP activity was higher in the testis and atrium, and lower in lung, in the OO group. Soluble GluAP activity was significantly lower in the testis of OO-fed animals. Membrane-bound GluAP activity did not differ between the two groups in any of the tissues analyzed. Serum AspAP and GluAP activities correlated negatively with palmitoleic and vaccenic acid respectively in the OO group. However, no significant correlations were found in the control group. These results may reflect functional changes in the renin-angiotensin system in the serum, adrenal, testis, lung and atrium after feeding with a diet enriched in olive oil.
Lipids have physical, chemical, and physiological properties that make them important factors in human nutrition. They form a group of compounds of varied chemical nature that have the common property of being soluble in organic solvents but insoluble in water. This basic property affects their digestion, absorption, and transport in the blood and metabolism at cellular level. Firstly, fatty-acid chain length and number of double bonds influence fat absorption. Thus, medium-chain fatty acids (MCFA) are better absorbed than long-chain fatty acids. Secondly, the positional distribution of fatty acids (FA) in dietary triglycerides (TG) determine whether FA are absorbed as 2-monoglycerides (2-MG) or free fatty acids (FFA), and hence, influences the composition of chylomicroms (CM) because triglycerides (TG) are resyntethysed in the intestinal mucosa using 2-MG from dietary lipids. Generally, the absorption of FA in the sn-2 position of TG is favored, whereas no specificity has been found for the fatty acids in the sn-1 and sn-3 positions. Finally, some FA of nutritional interest, namely, long-chain polyunsaturated fatty acids (LCP), are present in dietary lipid sources as both TG or phospholipids (PL). Fatty acids esterified as PL or TG may show different availability. In fact, some authors have suggested a better absorption of LCP-PL. Moreover, dietary LCP in form of TG or PL differently affects the composition of HDL and LDL PL.
Pyroglutamyl-ended forms of amyloid-beta-peptide are present in senile plaques in some individuals with Alzheimer type dementia. Single oral administration of the acetylcholinesterase inhibitor SDZ ENA 713 (rivastigmine (+)-(S)-N-ethyl-3-[(1-dimethylamino)ethyl]-N-methylphenylcarbamate hydrogen tartrate) increases basal and K(+)-stimulated pyrrolidone carboxyl peptidase (Pcp) activity in mice frontal cortex synaptosomes in a dose-dependent manner. These results suggest that this drug may ameliorate ATD cognitive deficits acting not only facilitating cholinergic transmission but also avoiding the formation of pyroglutamyl-ended amyloid-beta-peptides (A beta pE) deposition through the activation of Pcp.
The intake of mono- and polyunsaturated fatty acids has been associated with a minor risk of cardiovascular diseases including hypertension. Changes in levels of fatty acids may also modify the cell activity and may be related with alterations in different regulatory processes. Aminopeptidases are zinc-metalloenzymes which metabolise circulating peptide hormones in several tissues. Glutamyl-aminopeptidase (GluAP) and to a lesser extent, aspartyl-aminopeptidase (AspAP), are related with angiotensin metabolism in the renin-angiotensin system. The present work was designed to study the effect of a range of concentrations (1-100 microM) of oleic and linoleic acids and cholesterol present in the culture medium on the activity of GluAP and AspAP in the culture of rat cerebral cortical astrocytes taken from 21-day-old fetuses. The results showed that oleic acid inhibits, while linoleic acid stimulates the activity of AspAP. Both fatty acids inhibit the activity of GluAP. Cholesterol stimulates the activity of both enzymes. On the basis of these results, a functional link may exit between the effects of fatty acids on hypertension and the modulation of aminopeptidase activity by these compounds in rat astrocytes, as an example of target cell type in the central nervous system.
To evaluate the influence of light and darkness on brain pyroglutamyl-peptidase I (pGluPI) activity, four experimental groups of rats were compared at the same time-point (10.00 h). Two groups were designed with a standard 12-12 h light-dark cycle: In group A, the lights were on from 7.00 h to 19.00 h, and the experiment was done under light conditions; in group B, the lights were on from 19.00 h to 7.00 h, and the experiment was done under darkness conditions. Two additional groups were designed with nonstandard light-dark conditions: In group C, the animals were subjected to constant light, and the experiment was done under light conditions. In group D, animals were subjected to constant darkness, and the experiment was done under darkness conditions. Light (vs darkness) and standard (vs nonstandard) conditions produced significant changes on pGluPI activity in specific structures; the data suggested that endogenous substrates of pGluPI such as thyrotropin-releasing hormone and gonadotropin-releasing hormone, might be modified in parallel. There was left predominance in the retina under light conditions on a standard schedule (group A). The regional pattern of distribution of activity was similar in groups on a standard schedule (A vs B) and in groups tested under constant light-dark conditions (C vs D). However, this pattern differed between groups subjected to standard vs constant light-dark conditions (A and B vs C and D). These results support an influence of environmental light and darkness on pGluPI activity, which may reflect concomitant changes in its susceptible substrates and consequently in their functions.
The effect of killer strains of Saccharomyces cerevisiae on the growth of sensitive strains during must fermentation was studied by using a new method to monitor yeast populations. The capability of killer yeast strains to eliminate sensitive strains depends on the initial proportion of killer yeasts, the susceptibility of sensitive strains, and the treatment of the must. In sterile filtered must, an initial proportion of 2-6% of killer yeasts was responsible for protracted fermentation and suppression of isogenic sensitive strains. A more variable initial proportion was needed to get the same effect with non-isogenic strains. The suspended solids that remain in the must after cold-settling decreased killer toxin effect. The addition of bentonite to the must avoided protracted fermentation and the suppression of sensitive strains; however, the addition of yeast dietary nutrients with yeast cell walls did not, although it decreased fermentation lag.
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