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Biomedical subjects

G Arroyo

Publications and source records attributed to G Arroyo.

At least 19 recordsLinked to original sources

A phase I-II study of weekly cisplatin and gemcitabine with concurrent radiotherapy in locally advanced cervical carcinoma.

BACKGROUND: The purpose of this study was to determine the maximum-tolerated dose (MTD) and the antitumor activity of gemcitabine when administered in combination with concurrent cisplatin and radiotherapy in locally advanced cervical carcinoma (LACC). PATIENTS AND METHODS: Patients with histologically confirmed LACC (International Federation of Gynecology and Obstetrics IIB-IVA), previously untreated, were eligible for entry in the study to receive radiotherapy and concomitant weekly chemotherapy with cisplatin 40 mg/m(2) and gemcitabine at increasing doses levels until the MTD was found. RESULTS: Thirty-six patients were included. Sixteen patients were entered at four dose levels. The MTD was 150 mg/m(2) and the recommended dose of gemcitabine for phase II was 125 mg/m(2). Twenty additional patients were entered at this level. Toxicity at the recommended dose was acceptable with grade 3/4 toxicity in <20% of patients. Thirty-five of thirty-six patients (97.3%) achieved an objective response, 32 (88.8%) a complete response (CR) three a (8.3%) partial response and one (2.7%) stable disease. At a median follow-up of 26 months, 28 of 36 patients (77.7%) are in sustained complete remission and seven of 36 (19.4%) have relapsed. The 3-year disease-free and overall survival rates are 67% and 72%, respectively. CONCLUSION: The association of cisplatin and gemcitabine with concurrent radiotherapy is active and well-tolerated in untreated LACC.

Adult↗

Reaction of B. cereus bacteria and peroxidase enzymes under pressures >400 MPa.

It is known that B. cereus (Gram-positive bacteria) and peroxidase enzymes are resistant to pressures of approximately 400 MPa in fruit and vegetable products among others. The aim of the present work is to have knowledge about their behavior when using pressures >400 MPa without other combined treatments. The results showed that B. cereus was not inactivated at pressures of 1000 MPa for 15 min at 20 degrees C. In peroxidase enzymes the results remained similar and the pressure of 1000 MPa for 30 min was not enough to inactivate them. The apple cell structure at these high-pressure levels revealed that it changed and the cells were less cemented. The treated apple presented a translucent aspect, and some fluids migrated from the inside to the outside of the cell.

Bacillus cereus↗

Biological response of rats fed with tofu treated with high hydrostatic pressure.

Emerging technologies for food preservation have arisen in recent years, such as high-pressure (HP) hydrostatic treatment, and the biological response for this kind of food preservation is not well-known. Forty female rats (six weeks old) were used in the experiment to evaluate the biological effects of HP treatment of tofu. The animals were divided into groups that were fed with tofu (untreated), tofu treated with HP, and conventional food (as control) for 28 days. The glucose level, mineral content (calcium, potassium, zinc, and magnesium), shinbone maximum shear force, weight of the body, and weight of organs (heart, liver, spleen, and kidneys) were analyzed. The biological response for the rats was that significant differences were found in the calcium amount determined on the serum of the rats fed with untreated tofu and those fed with tofu treated with HP, and the calcium amount was lower on the rats fed with tofu treated with HP. Also, there were significant differences in the weight of the liver, and it was lower in the rats fed with tofu treated with HP. It was quite remarkable how the weight of the body and organs were smaller in the rats fed with tofu in comparison to the weight of the control rats. In the other components assayed no significant differences were found. HP produces a potential effect on tofu as it is observed in the rats response to the tofu treated with HP.

Animals↗

Soybean vegetable protein (Tofu) preserved with high pressure.

Tofu is a soybean vegetable protein that Asians have long consumed; its intake is increasing in other countries. Tofu was purchased at a local shop. The tofu samples were already preserved in plastic bags subjected to vacuum and storage (5 C). Tofu samples were subjected to high pressure (HP) of 400 MPa at 5 C for 5, 30, and 45 mm. Microbial analysis, sensorial evaluation, and structure were determined. HP treatment in tofu reduces the microbial population. Most of the microorganisms found in the initial population belonged to the Enterobacteriaceae family, bacteria Gram-negative (no Enterobacteriaceae), and bacteria Gram-positive. Alter HP treatment, Hafnia alvei and Bacillus cereus were found. After HP treatment, tofu is a pasteurized product, which is safer in terms of secondary pathogenic microbial contamination. Sensorial test results revealed that treated tofu was acceptable to consumers. The micrographs on the cryofracture observed with a cryoscanning electron microscope revealed a more compact structure after pressure compared with that of untreated samples, but the aggregates in the treated samples were more disperse.

Food Preservation↗

A spider tRNA(Ala) requires a far upstream sequence element for expression.

Within the series of timed differential accumulations of small RNAs we have shown to prelude the synthesis of fibroin in the large ampullate glands of Nephila clavipes (Nc), we are currently directing our attention to the alanine tRNAs. This work reports the subcloning of the members of a tRNAAla gene cluster and the optimization of their transcription in a heterologous cell-free system derived from Bombyx mori (Bm) silkglands. Our data show that the heterologous cell-free system supports the faithful and differential transcription of the individual spider alanine tRNA genes. We are thus making use of the extract to characterize the individual genes with respect to flank-contained regulatory elements through cell-free transcription of gene derivatives. The work has been initiated with pNTA3 because of its high transcriptional activity. Interestingly, the transcription of this gene requires a far upstream sequence, an uncommon modality in tRNA genes.

Animals↗

Protective effect of ascorbic acid against the browning developed in apple fruit treated with high hydrostatic pressure.

Apple Reineta variety was used as an apple dessert. The 1-1.5-cm cubes were immersed in a sucrose solution (30% w/v) and subjected to high pressure (HP) of 400 MPa for 30 min at 5 degrees C. Different ascorbic acid concentrations were used to protect the fruit from the browning developed after the HP treatment. After 2 months of storage at 5 degrees C, no brown color was observed in the samples treated with 20 mM ascorbic acid, and they were acceptable to consumers. However, untreated samples presented fermentation, and they were not acceptable to consumers. The electric conductivity and potassium content were found to be good indicators of the metabolites released from the fruit to the solution in samples treated with high pressure. HP did not affect the peroxidase activity but eliminated the microbial population.

Ascorbic Acid↗

High pressure response of fruit jams contaminated with Listeria monocytogenes.

High pressure is an alternative to thermal processing and is used to preserve food. Listeria monocytogenes is a bacterium which grows at low temperature, is able to multiply under vacuum, and is responsible for food poisoning. Pressures of 100, 200, 300 and 400 MPa were used for 5, 10 and 15 min at 20 degrees C on pure culture, and on apple and plum jam baby food artificially contaminated with Listeria. Pure culture was also to test pressures of 200, 300, 350 and 400 MPa at 5 degrees C for 30 min. The results were analysed statistically and showed that there were no significant differences between pressures of 100 and 200 MPa at 5, 10 and 15 min. However, at 300 MPa, there were significant differences at 15 min. When the pressure treatment was 400 MPa, significant differences were observed at pressure times of 5, 10 and 15 min. The results were fitted to a linear curve. In pure culture, no viable cells were detected after high pressure treatment of 350 MPa for 30 min at 5 degrees C. The use of low temperature helps to maintain the sensory properties of the product.

Colony Count, Microbial↗

Response to high-pressure, low-temperature treatment in vegetables: determination of survival rates of microbial populations using flow cytometry and detection of peroxidase activity using confocal microscopy.

Application of high hydrostatic pressure (200, 300, 350 and 400 MPa) at 5 degrees C for 30 min to different micro-organisms, including Gram-positive and Gram-negative bacteria, moulds and yeasts, proved to be more effective in inactivating these organisms than treatments at 20 degrees C for 10 min and at 10 degrees C for 20 min. Moulds, yeasts, Gram-negative bacteria and Listeria monocytogenes were most sensitive, and their populations were completely inactivated at pressures between 300 and 350 MPa. The same conditions of pressure, temperature, and time were applied to different vegetables (lettuce, tomato, asparagus, spinach, cauliflower and onion), achieving reductions of from 2-4 log units in both viable mesophiles and moulds and yeasts at pressures of between 300 and 400 MPa. Sensory characteristics were unaltered, especially in asparagus, onion, tomato and cauliflower, though slight browning was observed in cauliflower at 350 MPa. Flow cytometry was applied to certain of the microbial populations used in the above experiment before and after the pressurization treatment. The results were indicative of differing percentage survival rates depending on micro-organism type, with higher survival rates for Gram-positive bacteria, except L. monocytogenes, than in the other test micro-organisms. Growth of survivors was undetectable using the plate count method, suggesting that micro-organisms suffering from pressure stress were metabolically inactive though alive. The pressurization treatments did not inactivate the peroxidase responsible for browning in vegetables. Confocal microscopic examination of epidermal tissue from onion showed that the enzyme had been displaced to the cell interior. Use of low temperatures and moderately long pressurization times yielded improved inactivation of micro-organisms and better sensorial characteristics of the vegetables, and should lower industrial costs.

Aerobiosis↗

Effect of high pressure on the reduction of microbial populations in vegetables.

Resistance of micro-organisms to high pressure is variable and directly related to extrinsic and intrinsic factors. Pressures of 100, 200, 300, 350 and 400 MPa were applied at 20 degrees C for 10 min and at 10 degrees C for 20 min using strains of Gram-positive and Gram-negative bacteria, moulds and yeasts, as well as spores of Gram-positive bacteria. The results showed that at pressures of 100 and 200 MPa, decreases in microbial populations were not significant, whereas the populations of all the micro-organisms tested decreased considerably at a pressure of 300 MPa. A pressure of 300 MPa at 10 degrees C for 20 min was required to completely reduce the population of Saccharomyces cerevisiae, and a pressure of 350 MPa was needed to reduce most of the Gram-negative bacteria and moulds. The Gram-positive bacteria were more resistant, and pressures of 400 MPa were unable to completely reduce their populations. The different pressures employed had little effect on the initial numbers of spores. The initial populations of viable aerobic mesophiles and moulds and yeasts in vegetables (lettuce and tomatoes) decreased 1 log unit at pressures of 300 MPa and above under both sets of experimental treatment conditions. However, treatment at that pressure also resulted in alterations in the organoleptic properties of the samples. In the tomatoes, the skin loosened and peeled away, though the flesh remained firm, and colour and flavour were unchanged. The lettuce remained firm but underwent browning; flavour was unaffected. In vegetables use of moderate pressures in combination with other treatment conditions would appear to be required to reduce the populations of contaminating micro-organisms while avoiding the undesirable alterations in organoleptic properties that take place at 300 MPa.

Food Microbiology↗

Efficiency of different enrichment and isolation procedures for the detection of Salmonella serotypes in edible offal.

Rapid detection systems for Salmonella in foodstuffs are currently being developed. However, existing standards still call for application of traditional methods employing pre-enrichment followed by selective enrichment and isolation. The efficacy of various methods was tested using 264 chicken and lamb organ meats. Pre-enrichment was carried out in Tryptone Soy Broth (TSB) and enrichment in Tetrathionate Brilliant Green Broth (TTB) at 37 degrees C, Selenite Broth with Brilliant Green and Sulphapyridine at 37 degrees C and 43 degrees C, and Rappaport-Vassiliadis Broth (RV 10) at 42 degrees C. The isolation media were Brilliant Green Agar (BGA), Deoxycholate Citrate Agar, Hektoen Enteric Agar (HEA) and Salmonella-Shigella Agar. Enrichment in RV/42 degrees C followed by isolation on BGA as recommended by ISO standard no. 6579 and enrichment in TTB/37 degrees C followed by isolation in HEA, no longer recommended by that standard, produced the best results. Low percentages of positive samples and difficulties in detecting Salmonella are the result of interference by competing organisms (Enterobacteriaceae) and the number of salmonellas present after enrichment. A total of 528 samples (TSB, eggs, lamb liver and chicken liver) were inoculated with Salm. enteritidis, Salm. kapemba and Salm. virchow, and the preceding experiment was repeated. All the TSB and egg samples tested positive, but the percentage of positive samples from the lamb and chicken liver was only 81-92%. Recovery of the salmonellas did not depend upon the method employed or the serotype inoculated but instead on interference by competing flora and the numbers of Salmonella present in the samples.

Animals↗

Selective action of inhibitors used in different culture media on the competitive microflora of Salmonella.

The action of 12 inhibitors employed in the culture media used to detect the presence of Salmonella in food on 24 bacterial strains including contaminating Gram-positive bacteria common in water and food, Gram-negative bacteria, especially Enterobacteriaceae and Pseudomonadaceae, which are components of the competitive microflora, and six Salmonella serotypes was tested. Two liquid culture media (AR 5 and AE 1) were used. Series of tubes containing increasing concentrations of each inhibitor were inoculated with the test strains and incubated at 37 degrees C until growth was verified spectrophotometrically (24-48 h). The results showed that the inhibitors were effective against the Gram-positive contaminating microflora. They did not preferentially inhibit the competitive microflora of Salmonella, chiefly Enterobacteriaceae, and were ineffective against the Pseudomonas strains, which can tolerate concentrations higher than those customarily employed in culture media.

Anti-Bacterial Agents↗

Prelude activities in the synthesis of tissue-specific secretory protein products.

In studying the process of protein synthesis of a silk-producing organism we have found that several macromolecules must be synthesized in order for the process to occur. Through time course studies, we have found that small RNAs may play a paramount role in directing the finely orchestrated process. Alanine tRNA, U1 snRNA, and 5S RNA have been identified through Northern blotting as molecules timely and tissue-specific synthesized and upgraded as a prelude activity for the silk being produced.

Animals↗

Spider silkglands contain a tissue-specific alanine tRNA that accumulates in vitro in response to the stimulus for silk protein synthesis.

The large ampullate glands of the orb-web spider, Nephila clavipes provide massive amounts of fibroin throughout the lifetime of the adult female. We have developed methods to culture the glands and manipulate their biosynthetic activity. This has allowed us to monitor a series of molecular events that precede silk production in glands excised from appropriately stimulated animals. In this paper, we demonstrate that prior to the transient dramatic production of fibroin, such glands accumulate large amounts of tRNAs cognate to the abundant amino acids in spider silk. One of these, alanine tRNA, appears to consist of two isoaccepting forms--one constitutive, and the other silkgland specific. Moreover, the silkgland-specific form appears to accumulate preferentially in response to stimulation. This phenomenon of tissue-specific tRNA production appears similar to that found in the silkglands of Bombyx mori, but the spider system has the unique property of permitting manipulation in vitro. Thus, it provides an unusual opportunity to study the mechanism of regulated tRNA synthesis.

Animals↗

Changes in platelet aggregation caused by glyburide in diabetic patients.

The effect of glyburide on platelet aggregation was investigated in 31 patients with maturity-onset diabetes. Platelet aggregation in platelet-rich plasma was measured before and one hour after the patients had ingested 5 mg of glyburide. The drug caused a significant decrease in platelet aggregation as determined by changes in the amplitude and slope of the curve.

Adult↗