Dynamics of carotid body responses in vitro in the presence of CO2-HCO3-: role of carbonic anhydrase

The role of carbonic anhydrase (CNA) in the dynamics of carotid body (CB) function was tested by studying the effects of the membrane-permeable CNA inhibitor methazolamide on the chemosensory responses of the cat CB, perfused and superfused in vitro with cell-free and modified Tyrode solution at 36.5 +/- 0.5 degrees C in the presence of CO2-HCO3- (PO2 = 120 Torr, PCO2 = 32 Torr, pH = 7.40). The bulk of CO2 flow to the CB from the external milieu was overwhelmingly large relative to the metabolic production of CO2 in the CB. Accordingly, the relative contribution of the endogenous CO2 to the CB responses was small. The chemosensory nerve discharges were recorded from the whole desheathed carotid sinus nerve. The responses to acidic hypercapnia (PCO2 = 50-60 Torr, pH = 7.20-7.10), hypoxia (PO2 = 25 and 50 Torr), perfusate flow interruption, and bolus injections of sodium cyanide (20-40 nmol) were tested. To contrast, we also measured the effects of nicotine (2-4 nmol), which may act at sites other than those for O2 and CO2. Methazolamide (30 mg/l) in the perfusate at constant PCO2 and pH reduced the baseline activity and delayed the responses to step changes in PCO2 (and concomitantly pH) and PO2 and to cyanide but not to nicotine. The steady-state responses to these stimuli, measured as differences from control, were reduced, but not significantly. The initial overshoots seen with step changes in both high PCO2 and low PO2 were eliminated.(ABSTRACT TRUNCATED AT 250 WORDS)     

Electroclinical and neuroimaging studies in epilepsy

INTRODUCTION AND MATERIAL: During 54 months, we have studied the electro-clinical and neuroimaging features in outpatients with active epilepsy. Each patient was interviewed for one of us. Then, we have reviewed the medical records about both the clinical featuring. EEG and neuroimaging (NI) studies and seizures frequency (SF) outcome. Differences in crude proportions were assessed by chi 2 test for independence by 2 x 2 tables. RESULTS AND CONCLUSIONS: It has been 207 patients with 49 +/- 19.6 years of mean age at review. Partial seizures was significantly related with both a higher SF at onset and politherapy. Also, with a focal EEG distribution but only in case of complex partial seizures. Abnormal NI was significantly more frequent in oldest patients. A greater proportion of patients were in politherapy in four situation: SF at onset > 1 by day, a focal EEG distribution, duration of epilepsy longer than 20 years and age of onset lesser than 60 years. A 37.2% was seizures-free in the last year and in 34% the SF was improved a 50% or more from the beginning. A significantly greater proportion of patients was following with seizures in four cases: when the SF at onset has been > or = 1 by day, being partial seizures, women and having politherapy.     

Effect of inoculum size, relative humidity, storage temperature, and ripening stage on the attachment of Salmonella Montevideo to tomatoes and tomatillos

The influence of inoculum populations and environmental factors on attachment of Salmonella Montevideo to the surface of tomatoes and tomatillos was evaluated. To study the effect of inoculum size, red, ripe tomatoes were spot-inoculated with bacterial suspensions (10(5) and 10(8) CFU/fruit) and stored at 22 degrees C under 100% relative humidity. The effects of temperature (12, 22, and 30 degrees C) and relative humidity (75, 85, and 97%) on attachment of the pathogen (10(7) CFU/fruit) to tomatoes (red and green) and ripe tomatillos were also evaluated. Inoculated fruits were stored for 90 min at all combinations of temperature and relative humidity, and after rinsing with water, the number of cells attached to the surface was determined. Salmonella Montevideo attached to the surface of tomatoes within 90 min. A direct correlation between the number of attached cells and the population in the inoculum was observed. The percentage of cells that attached immediately after inoculation was approximately 0.3% for the three test products. After storage for 90 min at various temperature and relative humidity conditions, the number of adhering cells ranged from 4.0 to 5.4 log CFU/fruit (1.2% of inoculum). Both the type of product and the temperature/relative humidity combination had a significant (P < 0.05) effect on attachment of Salmonella Montevideo to the surfaces of tomatoes and tomatillos. Scanning electron micrographs of the cuticles of inoculated washed tomatoes and tomatillos revealed typical skin cell patterns, and only a few randomly dispersed Salmonella Montevideo were observed. Deposition of Salmonella Montevideo on the surface of tomatoes and tomatillos could result in attachment and subsequent colonization under suitable conditions.     

Trehalose Metabolism: From Osmoprotection to Signaling

Trehalose is a non-reducing disaccharide formed by two glucose molecules. It is widely distributed in Nature and has been isolated from certain species of bacteria, fungi, invertebrates and plants, which are capable of surviving in a dehydrated state for months or years and subsequently being revived after a few hours of being in contact with water. This disaccharide has many biotechnological applications, as its physicochemical properties allow it to be used to preserve foods, enzymes, vaccines, cells etc., in a dehydrated state at room temperature. One of the most striking findings a decade ago was the discovery of the genes involved in trehalose biosynthesis, present in a great number of organisms that do not accumulate trehalose to significant levels. In plants, this disaccharide has diverse functions and plays an essential role in various stages of development, for example in the formation of the embryo and in flowering. Trehalose also appears to be involved in the regulation of carbon metabolism and photosynthesis. Recently it has been discovered that this sugar plays an important role in plant-microorganism interactions.     

Cat carotid body chemosensory responses to non-hypoxic stimuli are inhibited by sodium nitroprusside in situ and in vitro

We studied the effects of sodium nitroprusside, a nitric oxide donor, on the chemosensory responses to cyanide and nicotine in the cat carotid body. In situ, sodium nitroprusside infusion reduced the cyanide-evoked responses in a dose-dependent manner. In vitro, Tyrode containing nitroprusside reversibly reduced the cyanide- (by 59%) and nicotine-induced (by 45%) chemosensory responses. The present results suggest that chemosensory responses induced by cyanide and nicotine are reduced by increased nitric oxide content, similarly to the hypoxic chemosensory responses.     

Carotid body chemoreception: the importance of CO2-HCO3- and carbonic anhydrase. (review)

The current hypotheses of carotid body (CB) chemoreception regard the glomus cells as the initial site of stimulus transduction. The consensus is that the transduction of chemical stimulus is coupled with the release of transmitter(s) from the glomus cells, which in turn generates action potentials in the afferent nerve terminals. Carbonic anhydrase (CA) is present in the glomus cells of the CB. Inhibition of CA activity in the CB in situ reduces the carotid chemosensory responses to CO2 and to O2, suggesting a common mechanism of chemosensing for both stimuli. However, CA inhibitors also block the red blood cell enzyme. Thus, the CO2 hydration reaction does not come to completion within the transit time of the blood from the lung to the CB. A steady-state reaction is not reached until later and so the PCO2 and pH levels in arterial blood samples are not the same as those sensed by the CB. Experiments in vitro using cat CB perfused and superfused with cell-free solutions, which had been pre-equilibrated with respiratory gases, strongly support the proposition that the CA activity in CB cells is essential for the speed and amplitude of the initial response to CO2 and for its subsequent adaptation. The immediate response to hypoxia also is delayed, but the late steady-state was less dependent on CA activity. In the nominal absence of CO2-HCO3- from the perfusate, hypoxic chemoreception persisted and its magnitude is not affected by CA inhibition, except for a delay which may be due to the initial alkaline pH of the glomus cells. Recent experiments performed in isolated glomus cells and in the whole CB show that hypoxia does not modify significantly the intracellular pH. By its simple catalytic function, CA can speed up the approach of the CO2 hydration reaction to equilibrium. However, CA may also contribute in the steady-state to the regulation of pHi by providing a continuous supply of H+ and HCO3-. Furthermore, CA performs a facilitatory role in the physiological chemosensory responses to CO2 and O2 in the presence of extracellular CO2-HCO3-. This role is likely to be related to the ion exchanger function and then to pHi regulation in the chemoreceptor cells.     

Time structure, temporal correlation and coherence of chemosensory impulses propagated through both carotid nerves in cats

In spontaneously breathing, pentobarbitone anesthetized cats, we recorded simultaneously the impulses in the chemosensory fibers of both carotid (sinus) nerves, to analyze the correlations between the frequencies of chemosensory discharges (f chi) and their activation (?df chi/dt?a) and deactivation (?df chi/dt?d) rates. We studied the chemosensory responses to brief exposures to hypoxia (100% N2; 5-s and 10-s) and hyperoxia (100% O2; 30-s), and intravenous injections of excitatory (NaCN 0.2-100 micrograms/kg) and inhibitory (dopamine hydrochloride 0.02-20 micrograms/kg) chemoreceptor agents. Hypoxia increased f chi, with a high temporal correlation between frequency levels in both nerves. Prolonging hypoxic stimulation increased ?df chi/dt?d, with preservation of ?df chi/dt?a. Hyperoxic exposure produced highly correlated decreases in f chi in both nerves, but reduced correlation in df chi/dt. Increasing doses of NaCN produced analogous increments in f chi, df chi/dt and their correlations, the ?df chi/dt?a/?df chi/dt?d ratio remaining constant along all the experimental range, except in one animal in which the ratio increased in both nerves alike. Dopamine reduced f chi bilaterally, with chemosensory silencing being reached with doses of about 0.2-0.5 microgram/kg, the correlations between f chi's of both nerves remaining constant within the range analyzed. Maximal ?df chi/dt?d was not affected along the range of dopamine doses, except in one animal in which it increased in both nerves. It is concluded that both carotid nerves convey similar quantitative information to the brain stem. Thus, the carotid nerves constitute either cooperative inputs or redundant afferences contributing to a high safety factor.     

Colonization of tomatoes by Salmonella montevideo is affected by relative humidity and storage temperature

The influences of the relative humidity (RH) and storage temperature on the colonization of tomato surfaces by Salmonella Montevideo were studied. Red, ripe tomatoes (Lycopersicon esculentum) were spot inoculated in three separate trials with 100 pl (approximately 10(6) CFU) of Salmonella Montevideo and stored for 90 min at 22 degrees C under 97% RH to facilitate attachment of cells to the blossom end of tomato surfaces. Following this attachment step, tomatoes were washed to remove loosely adhered cells and then stored at 22 or 30 degrees C for up to 10 days under RH of 60, 75, 85, or 97%. At 0, 0.4, 1, 4, 7, and 10 days of storage, three tomatoes were individually hand massaged in 50 ml of 0.1% peptone water and the washes were separately analyzed to enumerate populations of Salmonella Montevideo. The number of Salmonella Montevideo cells attached after 90 min at 22 degrees C was 3.8 log CFU per tomato; this level was determined to be the initial colonizing population. After 10 days of storage at 30 degrees C, the Salmonella Montevideo population increased to 0.7, 1.0, 1.2, and 2.2 log CFU per tomato at 60, 75, 85, and 97% RH, respectively. A similar trend was observed at 22 degrees C, although populations were lower than at 30 degrees C. Scanning electron micrographs of tomato cuticles after storage revealed a well-defined biofilm containing bacteria. These findings reinforce the importance of maintaining stored tomatoes at temperatures that do not support growth of pathogenic bacteria and demonstrate the growth-promoting effects of high humidity.     

A study of the retrogradation process in five argentine rice starches

Rate of the retrogradation process of five new argentine genotypes rice starches with different amylose contents and gelatinization temperatures during storage at 4.5 ± 0.5 °C was studied using differential scanning calorimetry (DSC).Significant differences were not found among transition temperatures, i.e., T₀ (onset temperature), T_p (peak temperature) and T_c (conclusion temperature), of non-waxy genotypes when studied using DSC. An increased retrogradation enthalpy (ΔH, mJ/mg) with the storage time occurred with all non-waxy samples. The waxy genotype, W4109, did not show a retrogradation peak for the period under study. Genotypes with high total amylose content (TAM) retrograded more than those with lower content. The kinetic parameters “k” and “n” were evaluated using the Avrami model appearing to be related to the water-soluble amylose (SAM) content. Relationship between the retrogradation degree (%R) and the water-insoluble amylose (IAM) content was found. Gelatinization temperature of the starch (T_G) seemed to affect the retrogradation rate.