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Biomedical subjects

J Gil

Publications and source records attributed to J Gil.

At least 127 records · Page 7Linked to original sources

Molecular cloning, expression, and chromosomal localization of a ubiquitously expressed human 6-phosphofructo-2-kinase/ fructose-2, 6-bisphosphatase gene (PFKFB3).

We report the identification of a human 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase gene (PFKFB3) isolated from a human fetal brain cDNA library. The gene was localized to 10p15-->p14 by fluorescence in situ hybridization. The entire cDNA (4,322 bp) codes for a polypeptide of 520 amino acid residues (molecular weight, 59.571 kDa). Structural analysis showed the presence of a kinase domain located at the amino terminus and a bisphosphatase domain at the carboxy terminus, characteristic of previously described 6-phosphofructo-2-kinase/fructose 2, 6-bisphosphatase isozymes. In addition, a phosphorylation site for cAMP-dependent protein kinase was found at the carboxy terminus. Northern blot analysis showed the presence of a unique 4.8-kb mRNA expressed in the different tissues studied. In mammalian COS-1 cells, this cDNA drives the expression of an active isozyme. Taken together, these results identify the presence of a gene coding for a human 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase isozyme which is ubiquitously expressed.

Animals↗

Alveolar vascularization of the lung in a lamb model of congenital diaphragmatic hernia.

Pulmonary hypoplasia and pulmonary hypertension are factors limiting the survival of infants with congenital diaphragmatic hernia (CDH). A reduction in the number of pre-acinar pulmonary vessels and increased muscularization are the structural lesions implicated as causes of irreversible hypoxemia. Whether there is a reduction in the number of air-blood barriers, which represent the capillary surface area of the lung involved in gas exchange, is unknown. We sought to determine if the lungs of CDH lambs have: (1) a reduction in total capillary surface area proportionate to the reduction in the total alveolar surface area of the lung; and/or (2) a disproportionate reduction in the number of capillaries (air blood barriers) within each acinus. The latter measurement was determined by calculating the capillary load which we defined as the number of air blood barriers/unit of surface density. At 80 d gestation (pseudoglandular period), a diaphragmatic hernia was created surgically in one lamb fetus of a twin gestation. At term, the fetuses were removed, the chests opened and the lungs fixed by a tracheal infusion of 1.5% glutaraldehyde at 25 cm of water pressure. Tissues from the lower lobes were examined by light and electron microscopy. Using computerized interactive morphometry, alveolar and capillary surface area, and capillary load were determined by intersection and point counting for the right and left lungs. The data show that the total alveolar surface area of the left CDH and control lungs were 1.8 +/- 0.8 m2 and 6.1 +/- 1.1 m2 (p < 0.01), respectively, and for the right CDH and control lungs 2.5 m2 +/- 0.1 and 11.2 +/- 1.9 m2 (p < 0.01), respectively. The total capillary surface area for the left CDH and control lungs were 0.7 +/- 0.3 m2 and 2.8 +/- 1.2 m2 (p < 0.05), respectively, and for the right CDH and control lungs 0.9 +/- 0.3 m2 and 3.8 +/- 1.5 m2 (p < 0.05), respectively. The capillary load was not statistically different. These findings demonstrate that the lungs in CDH are deficiently vascularized at the alveolar surface due to a reduction in the total alveolar surface area. Each acinus contains the same number of air blood barriers per unit of alveolar surface area indicating a normal acinar composition.

Animals↗

Organizing pneumonia in textile printing workers: a clinical description.

In April 1992 an outbreak of severe respiratory illness occurred among aerographic textile workers in the area of Alcoi, Autonomous Community of Valencia, Spain. An epidemiological study linked this outbreak to the use of a reformulated aerosolized product, Acramin-FWN. We analyzed clinical, laboratory, and pathological data of the first 14 patients with confirmed organizing pneumonia (OP) secondary to this newly recognized occupational toxicant. The mean age of the patients was 30 yrs. The most common clinical findings were cough (86%), epistaxis (71%), dyspnoea (64%), oppressive chest pain (57%), and crackles (50%). A restrictive functional pattern was evident in 64%. Radiographic findings consisted predominantly of patchy infiltrates in 65% and a micronodular pattern in 35%. Treatment with corticosteroids did not prevent initial progression in 11 of the 14 patients and development of irreversible respiratory failure in five patients. At necropsy, besides features of OP, interstitial fibrosis and diffuse alveolar damage were evident. A low total lung capacity, the presence of crackles at admission, and increases in the alveolar-arterial oxygen difference were predictive of death. The organizing pneumonia caused by the inhalation of Acramin-FWN is characterized by a tendency to evolve into progressive interstitial fibrosis despite the use of corticosteroids. The illness is restricted to the respiratory system and once respiratory failure has developed the prognosis is poor.

Adipates↗

Soluble and membrane-bound pyroglutamyl-peptidase I activity in the developing cerebellum and brain cortex.

Developmental changes of soluble and particulate pyroglutamyl (pGlu)-peptidase I activities in the rat brain cortex and in the cerebellum are described in this work. The enzyme activity has been measured spectrofluorimetrically using pGlu-b-naphthylamide as substrate (in the presence and absence of EDTA and DTT, necessary activators of the enzyme) in both soluble and particulate fractions. In the soluble fraction of the cerebellum and brain cortex, pGlu-peptidase I activity is high in the perinatal period and decreases two or three folds subsequently, at a later stage in the cerebellum than in the brain cortex, reaching adult levels at the end of the first postnatal month. The decrease in the activity of pGlu-peptidase observed in this work coincides with increasing levels of brain thyroliberin concentration after the second postnatal week. The particulate pGlu-peptidase I activity, obtained after osmotic shock and high-salt treatment, shows less significant changes during brain development in the areas under study. It is suggested that cytosolic pGlu-peptidase I could play a part in the normal development of the rat central nervous system.

Animals↗

[Transthoracic needle aspiration biopsy. The diagnostic accuracy of pulmonary lesions].

We performed 91 transthoracic functions, under fluoroscopy, sonographically or by computed tomography control, at Clinic Hospital in Valencia. The procedures was performed with Chiba needle 22 gauges while the cytopathologist was present. Maximum number of needle passes was four. Malignant diagnosis has been obtained in 64.8% of cases, in 12 patients a benign diagnosis was obtained and confirmed, 13 cases were false negative and the specimen was insufficient for diagnosis in 7 (5 of them lesions had a diameter less than 2 cm). The study showed 82% for sensitivity, 100% for specificity and 85% for accuracy. The punction was guided by fluoroscopy in 49 patients, by sonography in 6 and by computed tomography control in 36. The accuracy diagnosis for fluoroscopy guidance was 96%. The complications were 9 (9.9%): Two patients presented minor hemoptysis resolved spontaneously, 6 patients showed minor pneumothorax and one patient was treated a cause of a large pneumothorax. We concluded that transthoracic biopsy is a simple and save technique that can provide a high diagnosis accuracy in patients with pulmonary pathology.

Biopsy, Needle↗

Morphometric and electron-microscopic analyses of the effects of gonadotropin-releasing hormone agonists on uterine leiomyomas.

OBJECTIVE: Reports on the histologic effects of gonadotropin-releasing hormone agonists on uterine leiomyomas provide conflicting results. Most previous studies used qualitative methods of analysis. Using quantitative and semiquantitative stereologic methods of analysis, we assessed volume density of hyalinized areas, cell density, nuclear volume, and cytoplasmic cross-sectional areas of smooth muscle cells in histologic sections and also measured diameters of collagen fibrils in electron micrographs of uterine leiomyomas. DESIGN: Thirty leiomyomas from patients treated with gonadotropin-releasing hormone agonists (10 different patient samples), age-matched control patients (10 different patient samples), and postmenopausal women (10 different patient samples) were used. Hyalinization was assessed using a microscope with a projection head and affixed morphometric grid. Cell size and density were evaluated using a video-based, computerized system attached to the microscope, for which morphometric ad hoc programs were written. Diameters of collagen fibrils were measured from electron micrographs. SETTING: The study was conducted in the Department of Pathology, Mount Sinai Medical Center, New York, NY. PATIENTS: A total of 30 patient samples were studied, with three groups comprising 10 samples each, including patients treated with gonadotropin-releasing hormone agonists, age-matched control patients, and postmenopausal women. RESULTS: Myomas from patients treated with gonadotropin-releasing hormone agonists exhibited more hyalinization, greater cell density, slightly smaller cell sizes, and larger collagen fibrils than those of age-matched control patients and postmenopausal women. CONCLUSIONS: Shrinkage after treatment with gonadotropin-releasing hormone agonists is attributed to smaller cell size and increased collagenization in myomas.

Adult↗

Tuberculous pleurisy with or without radiographic evidence of pulmonary disease. Is there any difference?

SETTING: A community teaching hospital in Alicante, Spain. OBJECTIVE: To assess the characteristics of tuberculous pleurisy (TP) in our hospital, and to evaluate the differences between primary and reactivation forms. DESIGN: Between January 1984 and December 1993, all human immunodeficiency virus (HIV)-negative patients with TP were included in the study. From September 1987 onward, patients were prospectively studied. Charts, radiography, pleural fluid findings and diagnostic methods were evaluated. Two groups were distinguished according to chest radiographs: those patients with upper lobe lesions, calcified adenopathy and old pleural thickening were considered reactivation forms. RESULTS: Of the 129 patients (mean age, 31 +/- 18 years), 76% had primary TP and 24% reactivation TP. Differences were found in age (28 +/- 17 vs 40 +/- 18 years, P < 0.01), smoking (43% vs 74%, P < 0.01) and alcohol abuse (23% vs 47%, P < 0.05), weight loss (29% vs 50%, P < 0.05), positive sputum smears and cultures (2% vs 16%, 7% vs 28%, P < 0.01), and number of large effusions (46% vs 26%, P < 0.05), but not in tuberculin reactivity, pleural fluid findings, positive pleural cultures, or presence of pleural granuloma. CONCLUSION: In our setting, TP predominantly affects young adults. Clinical, immunological, and pleural findings are similar to those of patients with classic symptoms of TP. Older age, smoking and alcohol abuse, smaller effusions and sputum yield are differential characteristics of reactivation forms.

Adult↗

Involvement of CED-3/ICE proteases in the apoptosis of B-chronic lymphocytic leukemia cells.

B-chronic lymphocytic leukemia (B-CLL) is characterized by the accumulation of long-lived B lymphocytes that express high levels of Bcl-2. We examined the involvement of CED-3/ICE-like proteases in the apoptosis of B-CLL cells. One of the substrates of these proteases is poly(ADP [adenosine 5'-diphosphate]-ribose) polymerase (PARP). The effect of different factors that induce the apoptosis of B-CLL cells on the proteolytic cleavage of PARP has been studied. Treatment of B-CLL cells with different concentrations of dexamethasone (1 to 1,000 micromol/L) induced in a dose-dependent manner the cleavage of PARP. Dexamethasone induced PARP cleavage after 12 hours of incubation, which was almost complete at 48 hours. PARP cleavage during apoptosis of B-CLL cells was studied in cells from eight patients and a correlation was found between cell viability and the degree of PARP cleavage. Incubation in vitro of B-CLL cells with fludarabine for 48 hours induced PARP cleavage in all the cases studied. Protein kinase C (PKC) activation with 100 nmol/L TPA (12-O-tetradecanoylphorbol 13-acetate) or incubation with interleukin-4 (10 ng/mL) prevented either dexamethasone- or fludarabine-induced proteolysis of PARP. Incubation of B-CLL cells with the CED-3/ICE-like protease inhibitor Z-VAD.fmk inhibited spontaneous and dexamethasone-induced PARP cleavage and DNA fragmentation in a dose-dependent manner. Furthermore, Z-VAD.fmk prevented the cytotoxic effect of dexamethasone. These results indicate that CED-3/ICE-like proteases play an important role in the apoptosis of B-CLL cells.

Aged↗

Subcellular analysis of Tyr-aminopeptidase activities in the developing rat cerebellum.

The endogenous opioid system seems to play important roles in the developing cerebellum. The first opioid peptide isolated, Met-enkephalin, is expressed transiently in this brain area. In the present study, several enzyme activities capable of hydrolyzing enkephalins are measured during the first month of cerebellar development, using Tyr-beta-naphthylamyde as substrate and puromycin as inhibitor of one of the membrane-bound aminopeptidases. Puromycin-sensitive soluble and membrane-bound aminopeptidase activities decrease in the synaptosomal and mitochondrial fractions at the end of the first month of life, just when enkephalin-like immunoreactivity decreases in the cerebellum. Membrane-bound enzyme also decreases in the myelinic fraction. Synaptosomal activity increases after birth, coinciding with decreases in the activity in the microsomal fraction. Puromycin-insensitive and membrane-bound aminopeptidase shows less significant developmental changes and they occur mainly in the first week of life, coinciding with the axonal and dendrite growth. These results could suggest a possible role of these enzymes, together with the rest of the opioid system, in cerebellar development.

Aminopeptidases↗

Quantitative alterations of the functionally distinct subsets of CD4 and CD8 T lymphocytes in asymptomatic HIV infection: changes in the expression of CD45RO, CD45RA, CD11b, CD38, HLA-DR, and CD25 antigens.

We determined the representation in asymptomatic human immunodeficiency virus (HIV) infection of the CD45RO+ and CD45RO- CD45RA+ subsets of CD4+ and CD8+ T lymphocytes, CD11b+ and CD11b- subsets of CD8+ T cells, and activated populations of these subsets. Three-color flow cytometry was used to quantitate the different CD4+ and CD8+ T cell populations in 116 asymptomatic HIV+ individuals. In asymptomatic HIV+ infection there was a significant relative increase in the CD4+ CD45RO+ and CD8+ CD45RO+ T cell subsets, which express CD38 and DR antigens, that correlated strongly with the decline in total CD4+ T cells. In addition, we found a loss of CD4+ CD45RO- and CD8+ CD45RO- T cells associated with progression of HIV infection (as measured by the decline in total CD4+ T cells). Studies presented here also indicate that, with the progression of asymptomatic HIV infection, CD8+ CD11b- T lymphocytes showed a significant decrease, whereas CD8+ CD11b+ T cells were significantly increased. This study demonstrates that the progression of HIV infection in asymptomatic patients involves the increase in CD45RO+ subsets of CD4+ and CD8+ T cells, the increase in CD8+ CD11b+ T cells, the decrease in CD45RO- CD45RA+ subsets of CD4 and CD8 T cells, and the decline in CD8+ CD11b- T cells.

ADP-ribosyl Cyclase↗

Effect of growth factors on the expression of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in Rat-1 fibroblasts.

The activation of glycolytic flux is a biochemical characteristic of growing cells. Several reports have demonstrated the role of fructose 2,6-bisphosphate in this process. In this paper we show that the levels of 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase (6PF2K/Fru-2,6-P2ase) mRNA are modulated in response to serum and growth factors and this effect is due to regulation of its transcription rate. The modulation of the expression of this enzyme by growth factors differs according their mitogenic effect; both lysophosphatidic acid and epidermal growth factor, when added alone, increased the mRNA levels, but endothelin had no effect. Furthermore, cAMP, which acts as an antimitogenic signal in Rat-1 fibroblasts, produced a decrease in 6PF2K/Fru-2, 6-P2ase mRNA and inhibited the effects of lysophosphatidic acid and epidermal growth factor on 6PF2K/Fru-2,6-P2ase expression. PD 098059, a specific inhibitor of the activation of the mitogen-activated protein kinase, was able to prevent the effect of EGF on 6PF2K/Fru-2, 6-P2ase gene expression. These results imply that activation of mitogen-activated protein kinase is required for the stimulation of the transcription of 6PF2K/Fru-2,6-P2ase by EGF.

Animals↗

Morphometric study of the placental vessels and its correlation with umbilical artery Doppler flow.

OBJECTIVE: To determine the changes in the vessel-wall thickness and the radius of the lumen in tertiary-stem villi of the placenta with advancing gestational duration and their relationship to umbilical artery Doppler flow studies. METHODS: Placentas from 63 miscarriages and preterm and term deliveries (between 19 and 40 weeks) were used for morphometric study of the tertiary-stem villi vessels. Each woman had undergone Doppler flow study of the umbilical artery. The resistance index (RI) was determined from the Doppler flow velocity waveform. Placental paraffin sections of 4-micron thickness were stained with hematoxylin and eosin and with periodic acid-Schiff reagents. The tertiary-stem villi and their vessels were examined microscopically and assessed morphometrically using a personal computer with math co-processor and a touch-sensitive screen overlying a video monitor. The monitor received microscopic images from a video camera that was mounted on a microscope. We determined vessel-wall thickness by tracing the outer and inner circumferences of digitized vessel-wall images. RESULTS: Wall thickness, but not lumen size, of the tertiary-stem villi vessels decreased significantly overall at a rate of 0.63micron/week (P < .001). The rate of decrease was 0.64micron/week (P < .001) during the second trimester and 0.50micron/week (P < .001) during the third trimester. There was a significant correlation between the decrease in thickness and in RI (r = 0.83 [P < .001], r = 0.78 [P < .001] in the second and third trimesters, respectively). Resistance indices were all within normal limits. CONCLUSIONS: Placental tertiary-stem villi vessel-wall thickness decreases with advancing gestational age. There is a correlation between the changes in RI of the umbilical artery Doppler flow and the changes in mean wall thickness of the placental vessels.

Female↗

Effect of lidocaine administration on the enkephalin-degrading aminopeptidase activities in discrete areas of the rat brain.

1. The levels of three aminopeptidase activities involved in the degradation of enkephalins (soluble, M and MII) in several rat brain areas were studied after lidocaine administration. 2. Soluble aminopeptidase activity showed decreases in the frontal cortex and in the pituitary gland after treatment. 3. No significant changes in the complete membrane-bound aminopeptidase activity were appreciated in any other of the brain areas. 4. However, decreases of the membrane-bound puromycin-insensitive aminopeptidase activity in the frontal cortex, the hippocampus and the thalamus, after lidocaine administration, were observed.

Aminopeptidases↗

Reproducibility and accuracy of interactive segmentation procedures for image analysis in cytology.

The segmentation of nuclear images is a crucial step in the development of procedures using image analysis for the cytological diagnosis of cancer. The purpose of this study is to evaluate the reproducibility and accuracy of several interactive segmentation methods which can be used in this context. Four methods were studied: a thresholding-based method enabling selection of intensity histogram contrast and brightness, manual tracing with a stylus, and arc- and ellipse-fitting routines. Features of nuclear size and shape were derived from nuclei segmented on repeated occasions by several individuals. Variance component models provided a statistical framework for evaluating the intraobserver and interobserver variability of these measurements in terms of their intraclass correlation coefficients. Of the methods tested, the arc-fitting segmentation method gave the most reproducible results, and thresholding the least. Reproducibility was generally very high both between individuals and for repeated segmentations by a single individual. Accuracies of area measurements for the various methods, as determined with respect to point counting, paralleled the reproducibilities of the methods. Sample size requirements were observed to be more dependent on the biological variability of the tissue sampled than on the particular segmentation method or on the number of individuals performing segmentation.

Cell Biology↗

Endoscopic sphincterotomy for choledocholithiasis: a prospective single-center study on the short-term and long-term treatment results in 483 patients.

BACKGROUND AND STUDY AIMS: Endoscopic sphincterotomy (EST) is the method of choice for the treatment of choledocholithiasis. The aim of the present study was to assess the impact of various prognostic factors on the short-term and long-term results of EST. PATIENTS AND METHODS: Data from 483 EST procedures carried out due to choledocholithiasis without concomitant neoplasms or hepatic cirrhosis, and without subsequent bile duct surgery, were analyzed prospectively. The 324 patients (67%) who agreed to take part in the follow-up examinations and fulfilled the inclusion criteria for the study were divided into four groups, depending on the presence of the gallbladder or cholecystolithiasis prior to EST and on the time of diagnosis, and were prospectively followed up by physical examination laboratory tests, abdominal ultrasound, and endoscopic retrograde cholangiography examinations to evaluate the short-term and long-term outcomes. RESULTS: EST was successful in 95.7% of cases. Early complications were noted in 6.8% of cases. No early complications were seen in the group after cholecystectomy with a T-tube present, whereas this rate was highest in the group with concurrent choledocholithiasis and cholecystolithiasis (six of 48, 12.5%). Recurrent choledocholithiasis was observed in 5.6% of the cases overall. This rate was at its lowest in patients with choledocholithiasis alone, and was highest in the group with concurrent choledocholithiasis and cholecystolithiasis. Among the latter patients, the highest rates of papillary stenosis, chronic pancreatitis, and cholangitis were also observed, the latter complication being noted only in cases of recurrent common bile duct stones. CONCLUSIONS: EST is a relatively safe and effective procedure in the treatment of choledocholithiasis. The best prognostic factor is the presence of choledocholithiasis alone. Concurrent choledocholithiasis and cholecystolithiasis carry a more adverse prognosis, and in these cases cholecystectomy should be considered after EST.

Abdominal Pain↗