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Biomedical subjects

J A Oliveira

Publications and source records attributed to J A Oliveira.

At least 19 recordsLinked to original sources

Morphology and preliminary enzyme characterization of the salivary glands from the predatory bug Podisus nigrispinus (Heteroptera: Pentatomidae).

Podisus nigrispinus (Dallas) is a common predator in agricultural and natural systems in Neotropical America. Its feeding strategy involves extra-oral digestion and to better understand this process its salivary glands were extracted and subjected to morphological and preliminary enzyme characterization. The salivary glands of P. nigrispinus are formed by a pair of main and accessory gland complexes. The main salivary glands are further divided into an anterior and a posterior lobe. The compartmentalization of the salivary gland complex is likely to be important for the production, activation and release of the digestive enzymes used in the extra-oral digestion of prey items. Proteases and lipase, important digestive enzymes involved in zoophagy, were detected in the salivary glands of P. nigrispinus. The prevailing trypsin-like protease activity was characterized by using the serine-protease substrate N-alpha-benzoyl-L-Arg-p-nitroanilidine (L-BApNA) and the trypsin inhibitors tosyl-L-lysine chloromethyl ketone (TLCK) and benzamidine. The KM value obtained for trypsin-like activity was 1.57 mm and the different peaks of optimum pH and temperature activity suggest the presence of multiple forms of this enzyme in P. nigrispinus. Detection of amylase activity in the salivary glands of this predator suggests its ability to digest starch and obtain nutrients from plants, which may have adaptative value under prey scarcity.

Amylases↗

Detection of a chloramphenicol efflux system in Escherichia coli isolated from poultry carcass.

An active chloramphenicol efflux system was demonstrated in a multiresistant E. coli isolated from poultry carcass. The effect of different concentrations of chloramphenicol on the original strain and on the plasmid-cured strain was determined in the presence and in the absence of CCCP, an uncoupler of the proton-motive force. Minimal inhibitory concentration (MIC) was lower in the presence of CCCP in the original strain. The plasmid-cured strain displayed lower resistance for chloramphenicol than the wild type, but the MIC was not affected by CCCP. The combined results indicate a plasmid encoded energy dependent resistance mechanism. 3H-chloramphenicol accumulation within the cells was measured by scintillation counting. The uptake or the efflux of 3H-chloramphenicol was influenced by CCCP in the original strain, but not in the plasmid-cured strain. More than one chloramphenicol resistance mechanism may exist in this strain. E. coli is an important commensal or pathogen that inhabits the gastrointestinal tracts of humans and animals, so a plasmid encoded active drug resistance mechanism can be a potential source of horizontal transfer of resistance.

Animals↗

Postprandial symptoms in dysmotility-like functional dyspepsia are not related to disturbances of gastric myoelectrical activity.

Gastric dysrhythmias, such as tachy- or bradygastria, have been reported in patients with functional dyspepsia (FD), but their role in symptom production is uncertain. It is also not known whether gastric dysrhythmias in these patients can be elicited by physiological gastric distension with a meal. We investigated the relationships between symptoms after ingestion of different volumes of water following a test meal and gastric dysrhythmias in FD patients. Fourteen patients with dysmotility-like FD and 13 healthy volunteers underwent paired electrogastrography (EGG) studies. Fasted subjects ingested 150 ml of yoghurt with either 150 ml (low volume) or 300 ml (high volume) water in random order. Fasting and fed EGGs with monitoring of symptoms were performed in both studies. Ten FD patients (71.4%) reported upper abdominal discomfort and bloating after the low volume meal, but only one (7.1%) presented an abnormal EGG (dominant frequency in the 2-4-cpm range: 58%). Following the high volume meal, 7 patients (50%) had symptoms, but none had EGG abnormalities. No significant differences were found between FD patients and controls for any of the EGG variables, in any test. In FD patients with postprandial symptoms, the percentage of the EGG dominant frequency in the normal range (median, 84.6%; range, 76.0-100.0%) was similar (P>0.20) to that in those without symptoms (88.5%; 75.0-100.0%). We conclude that disturbances of gastric myoelectrical activity are unlikely to play a role in the origin of postprandial upper abdominal discomfort and bloating in dysmotility-like FD.

Adult↗

Efficacy of linear and convex transducers for ultrasound-guided transvaginal follicle aspiration.

A new device was developed to hold linear transducers for transvaginal follicle aspiration. Efficacy of follicle aspiration was compared using a linear 6 MHz and a convex 5 MHz transducer. Fifty-five cows were submitted to follicle aspiration at random days of the estrous cycle. Aspirations were conducted with linear (n=28) and convex (n=38) transducers with 18 G needles at a negative pressure corresponding to 13 ml H(2)O/min. A greater number of follicles were aspirated using convex than to linear probe (12.4 versus 7.8, respectively, P<0.05). Mean number of oocytes and recovery rates were similar for convex (5.4 and 48.6%) and linear (4.6 and 59.3%) transducers. Limited space between the linear transducer and needle guide restricted access to some portions of the ovary, reducing the number of follicles aspirated using a linear transducer. The newly developed adaptor allowed greater stability, holding the ovaries firmly against the linear transducer. This diminished mobility permitted a similar number of oocytes to be recovered with both transducers. In conclusion, this new adaptor provided a low cost alternative for routine follicle aspiration and oocyte recovery in cattle.

Animals↗

Molecular and karyologic variation in the genus Isothrix (Rodentia, Echimyidae).

Phylogenetic relationships among available samples of the genus Isothrix using mitocondrial gene cytochrome b were carried out and a new karyotype of Isothrix specimens referred to Isothrix negrensis from the mid course of Rio Negro, in the Brazilian Amazon, was described. Molecular, karyological and morphological data of Isothrix negrensis was compared to that of I. bistriata and I. pagurus. Cytochrome b DNA sequence analyses clearly separate I. negrensis from I. bistriata and genetic distances estimates between then are greater than between species of the related genus Mesomys. The median-joining analysis postulated the presence of two median vectors between I. bistriata haplotypes from adjacent localities, suggesting that genetic isolation between them is unrelated to geographic distance. These results confirm previous molecular differences suggesting that I.bistriata is a composite of several taxa. The karyotype of Isothrix negrensis also differs from those reported for I. bistriata and I. pagurus. In relation to the external morphology I. negrensis differs from I. bistriata by the overall darker dorsal coloration, head darker than dorsum, presence of a longitudinal line of blackish hairs in the proximal part of the tail extending until near to its base, dorsal surface of hind feet covered with yellowish hairs and stripe of light hairs in the head reduced to a small patch.

Animals↗

Histoenzymology and morphometry of the masticatory muscles of tufted capuchin monkey (Cebus apella Linnaeus, 1758).

Samples of the anterior and posterior regions of the masseter and temporal muscles and of the anterior belly of the digastric muscle of 4 adult male tufted capuchin monkeys (Cebus apella) were removed and stained with HE and submitted to the m-ATPase reaction (with alkaline and acid preincubation) and to the NADH-TR and SDH reactions. The results of the histoenzymologic reactions were similar, except for acid reversal which did not occur in fibers of the fast glycolytic (FG) type in the mandibular locomotor muscles. FG fibers had a larger area and were more frequent in all regions studied. No significant differences in frequency or area of each fiber type were detected, considering the anterior and posterior regions of the masseter and temporal muscles. The frequency of fibers of the fast oxidative glycolytic (FOG) and slow oxidative (SO) types and of FOG area differed significantly between the anterior belly of the digastric muscle and the mandibular locomotor muscle. The predominance of fast twitch (FG and FOG) fibers and the multipenniform and bipenniform internal architecture of the masseter and temporal muscles, respectively, are characteristics that permit the powerful bite typical of tufted capuchin monkeys.

Adenosine Triphosphatases↗

Relationship between follicle size and ultrasound-guided transvaginal oocyte recovery.

We evaluated the relationship between follicle size and oocyte recovery (OR) using ultrasound-guided follicle aspiration. Thirty Holstein cows were subjected to OR without gonadotrophic therapy. Oocytes were recovered two to four times from each cow in a total of 67 aspiration sessions. Ovarian follicles with diameters < or =4 mm and >4 mm were aspirated in separated groups. Recovered oocytes from each group were kept separate and submitted to in vitro maturation, fertilization, and culture to the blastocyst stage. A total of 430 follicles were aspirated, of which 154 (35.8%) were from follicles >4 mm and 276 (64.2%) were from follicles < or =4 mm. Seventy-seven oocytes (50%) were recovered from follicles >4 mm and 200 (72.2%) were from follicles < or =4 mm. Nineteen blastocysts were obtained from follicles >4 mm, whereas 45 blastocysts were obtained from follicles < or =4 mm. Recovery rate was greater (P<0.01) in follicles < or =4 mm. Oocyte quality, cleavage rate and blastocyst development did not differ between different follicle sizes. Routine aspiration of small follicles (< or =4 mm) could increase the number of oocytes available for in vitro development.

Animals↗

Alcohol determination using an acoustic wave sensor.

An acoustic wave methodology was developed to quantify alcohols in aqueous solutions. The frequency at minimum impedance of a bare quartz crystal in contact with ethanol solutions was found to be a suitable parameter to quantify ethanol. Ethanol in several Portuguese white wines was analyzed both by the proposed methodology and by the usual areometric method with no statistically significant differences (alpha = 0.05) in precision or accuracy of the results.

Electrochemistry↗

Development of a sensor for calcium based on quartz crystal microbalance.

A new sensor for calcium based on a piezoelectric quartz crystal is presented. The selectivity depends on the ratio lipophilic salt/ionophore of the composition of the coating of the quartz crystal. A crystal coated with a THF solution of PVC (34.5% w/w), DOS (62.1% w/w) and 10,19-bis[(octadecylcarbamoyl) methoxyacetyl]-1,4,7,13,16-pentaoxa-10,19-diaza cycloheneicosane (3.4% w/w) and a salt/ionophore molar proportion of 60%, corresponding to a frequency decrease of the dry crystal of 6.0 kHz, showed a detection limit to calcium of 2.2 mg/L. Both a standard calcium chloride solution and a commercially balanced salt solution for tissue culture were analyzed subsequent to ion chromatographic separation. The results obtained with the sensor developed were compared with those obtained with a conductivity detector. Although the results from both detectors agreed for the standard solution, only the quartz crystal sensor was able to give reliable results for the tissue culture solution.

Calcium↗

[Time's paradigm in primary angioplasty].

The use of primary angioplasty in the treatment of acute myocardial infarction (AMI), as opposed to fibrinolysis, is well established, as long as performed by an experienced team. The benefits are "time-dependent", hense the rational for the sentence "Time is Muscle". It is very important to assess the time factor, mainly if the therapeutic option is primary angioplasty, since it is necessary to set up a full range of conditions related to this type of specific treatment (human, material and logistical). In the current study was analysed the in and out of hospital time sequence of 99 patients with AMI that underwent primary PTCA during the year of 1999. The main goal was to assess if the performance of this procedure outside regular working hours could have any significant impact on the "time of in-hospital ischemia". The different time durations were prospectively recorded according to a protocol that was previously established and filled for each patient by the lab team. The authors conclude that: a) the average in-hospital time delay in the treatment of AMI by PTCA was not significantly influenced by the fact that the procedure was performed outside the regular working hours of the lab; b) the main component of the "ischemic in hospital time" corresponds to the time between the patient's admission and the decision making time for PTCA. Although the average decision making time for PTCA is too long and must be optimized, the average total in hospital ischemic time (99-105 minutes) compares favorably with other previous reports in the literature.

Aged↗

The quantification of potassium using a quartz crystal microbalance.

N,N'-dibenzyl-4,13-diaza-18-crown-6 (A) and bis[(benzo-15-crown-5)-4'-ylmethyl] pimelate (B) were tested as coatings for two piezoelectric crystals for potassium quantification. Both sensors showed stability, reversibility and sensitivity characteristics that allowed their use in quantitative analysis. However, compound A is much more sensitive to potassium than B. Compound A also shows a larger relative sensitivity for potassium with regard to sodium than B. A pharmaceutical sample of known composition was analysed both by an acoustic wave sensor with a crystal coated with compound A and by conductivity. No statistically significant difference in the median of the results was found (alpha = 0.05), although precision is superior for the conductivity methodology. Performance of the sensor in terms of frequency stability and selectivity was improved by the incorporation of PVC, a plasticizer and a lipophilic salt in the coating composition. Limits of detection found for potassium were 1.92 ppm, or 1.75 ppm for a crystal with a frequency decrease due to coating of 2.9 kHz or 3.9 kHz, respectively. Selectivity coefficients (fixed interference 80 ppm) for potassium over Na, Ca, Al, Zn, Mg, and Fe ranged from 0.103 to 0.332.

Flow Injection Analysis↗

DNA release by line-1 (L1) retrotransposon. Could it be possible?

We have verified the presence of line-1 retrotransposon (L1) in plasma DNA in 15/17 brain tumor (glioma) patients and in 6/6 healthy people by applying PCR amplification of part of the L1 5' end. The same samples were separately amplified for K-ras. Results suggested that L1 sequences are circulating throughout the body. We hypothesized the participation of transposable elements such as L1 in a putative DNA release mechanism.

Adult↗

Trends in alkanes and PAHs in airborne particulate matter from Oporto and Vienna: identification and comparison.

A total of 56 weekly samples from Oporto and 40 from Vienna were collected and analysed, for 23 n-alkanes and 16 polycyclic aromatic hydrocarbons (PAH), by GC-MS after extraction with a toluene/methanol mixture. Total and elemental carbon were in the same range of values for both sampling sites. Although parts of the spectrum of species in both sampling sites were constant over the sampling period there is no evidence for suggesting a universal tracer for alkanes. For both Oporto and Vienna, Principal Component Analysis (PCA) identified two PCs and in both cases the first PC contained only PAHs while the second contained only alkanes. This separation between alkanes and PAHs and the observation of an alternating pattern with higher concentrations of odd carbon numbered from C27 to C30 (natural emissions from biological origin) is believed to result from a separation between anthropogenic and biological contributions associated with the first and second PC, respectively.

Air Pollutants↗

A new method for monitoring the contamination of glycerol with diethylene glycol.

A quartz crystal microbalance has been used to check the purity of glycerol. The experimental procedure involves no derivatization or other preliminary step except determination of water content. The protocol consists of the direct reading of the frequency of a quartz crystal with one face in contact with an aqueous solution of the glycerol to be tested. The method is inexpensive, simple, rapid, and highly sensitive; diethylene glycol contamination at the 3.61% level could be detected.

Biosensing Techniques↗

Reduced exploratory activity of audiogenic seizures susceptible Wistar rats.

Patients with epilepsy and animals with experimental usually show behavioral changes such as increased anxiety. Audiogenic seizures (AS) are a model of generalized tonic-clonic limbic seizures induced by sound stimulation in genetically susceptible animals. The objective of this paper was to evaluate the exploratory activity of an inbred strain derived from Wistar progenitors that has been selected in our laboratory for AS susceptibility. The exploratory activity of audiogenic seizures susceptible (S) and resistant (R) Wistar rats was measured in two situations: an open arena and the elevated plus maze, an animal model of anxiety. S animals displayed a reduced exploration in both the open arena (reduced total distance moved) and the elevated plus maze (reduced number of enclosed-arm entries). In the latter there was also a decrease in open-arm exploration, particularly of the distal part of these arms. This effect persists even when the effect of a decreased number of enclosed-arm entries is removed by analysis of covariance. Therefore, the results indicate that audiogenic seizure genetically susceptible Wistar rats display a reduced exploration of novel environments. Moreover, the results with the elevated plus maze suggest that these animals are more anxious than AS resistant rats.

Acoustic Stimulation↗

Optimisation of the experimental conditions of a new method, based on a quartz crystal microbalance, for the determination of cyanide.

A new method based on a quartz crystal microbalance was developed for the determination of cyanide. As the sensitivity depends on pH, temperature and nitrogen flow rate, a modified simplex was used to optimise these experimental parameters. Two different versions of the proposed method were optimised. For the first version a sensitivity increase of 1.5 was observed after 27 runs, whereas for the second version a sensitivity increase of 1.7 was observed after 12 runs.

Cyanides↗

Anticonvulsant and proconvulsant roles of nitric oxide in experimental epilepsy models.

The effect of acute (120 mg/kg) and chronic (25 mg/kg, twice a day, for 4 days) intraperitonial injection of the nitric oxide (NO) synthase (NOS) inhibitor NG-nitro-L-arginine (L-NOARG) was evaluated on seizure induction by drugs such as pilocarpine and pentylenetetrazole (PTZ) and by sound stimulation of audiogenic seizure-resistant (R) and audiogenic seizure-susceptible (S) rats. Seizures were elicited by a subconvulsant dose of pilocarpine (100 mg/kg) only after NOS inhibition. NOS inhibition also simultaneously potentiated the severity of PTZ-induced limbic seizures (60 mg/kg) and protected against PTZ-induced tonic seizures (80 mg/kg). The audiogenic seizure susceptibility of S or R rats did not change after similar treatments. In conclusion, proconvulsant effects of NOS inhibition are suggested to occur in the pilocarpine model and in the limbic components of PTZ-induced seizures, while an anticonvulsant role is suggested for the tonic seizures induced by higher doses of PTZ, revealing inhibitor-specific interactions with convulsant dose and also confirming the hypothesis that the effects of NOS inhibitors vary with the model of seizure.

Animals↗