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Biomedical subjects

L Campos

Publications and source records attributed to L Campos.

At least 19 recordsLinked to original sources

Restricted diet modifies carbohydrate metabolism in immature rats.

The aim of the present work was to study the effect of a restricted diet on carbohydrate metabolism in submandibular glands of female prepuber rats and the influence of arachidonic acid derivatives. Rats of 21 days of age were divided into three groups. Group I: normally fed rats. Group II: restricted diet (50% of the normal food intake). Group III: normally restricted diet with re-feeding. The baseline concentration of tissue glycogen was significantly higher in Group II than in I or III and after 60 min of incubation in a glucose free medium all groups showed a glycogen drop. In addition, the glucose metabolism was increased in Group II. Cycloxigenase inhibitors failed to alter (14)CO(2) levels in Groups I and III. In Group II, indomethacin and acetylsalicylic acid inhibited glucose metabolism, which was reverted by PGE(2) addition. The exogenous arachidonic acid metabolism and different eicosanoids showed that restricted diet significantly increased the production of PGE(2) but diminished PGF(2)(alpha) production. Our results suggest that a restricted diet would lead to a new dynamic equilibrium of glucose homeostasis. Prostaglandins E(2) and F(2)(alpha) would participate by adapting the source of energy to tissue demands while maintaining the metabolic features that characterize puberty.

Animals↗

A randomized, multicenter study of subcutaneous and intravenous darbepoetin alfa for the treatment of chemotherapy-induced anemia.

BACKGROUND: This randomized, open-label study evaluated the efficacy, safety and pharmacokinetics of darbepoetin alfa administered intravenously (i.v.) or subcutaneously (s.c.) in chemotherapy-induced anemia. PATIENTS AND METHODS: Patients received darbepoetin alfa i.v. (n=59) or s.c. (n=59) at a dose of 4.5 mug/kg once weekly for 6 weeks (correction phase) followed by 4.5 mug/kg once every 3 weeks for the remainder of the 18-week treatment period (maintenance phase). RESULTS: During the correction phase, the mean [95% confidence interval (CI)] change in hemoglobin (intention-to-treat) was 1.1 (0.6-1.5) g/dl in the i.v. group and 1.3 (0.9-1.7) g/dl in the s.c. group; using available data, the mean change was 1.4 (1-1.9) g/dl and 1.6 (1.2-2) g/dl, respectively. The percentage (95% CI) of patients maintaining hemoglobin (i.e. average decrease < or =0.5 g/dl) during the maintenance phase was similar between the i.v. (82%; 95% CI 66% to 92%) and s.c. (80%; 95% CI 66% to 90%) groups. Thirty-five per cent (95% CI 20% to 50%) of patients in the i.v. group and 32% of patients in the s.c. group (95% CI 18% to 45%) received red blood cell transfusions during week 5 to the end of the treatment period. Darbepoetin alfa was well tolerated in both groups. No significant difference (P=0.36) in weekly darbepoetin alfa serum concentrations was observed between groups. CONCLUSIONS: Darbepoetin alfa can be administered i.v. or s.c. at equal doses for the treatment of anemia in this setting.

Aged↗

Synthesis and secondary structure of oligo(alpha-isobutyl beta,L-aspartate)s.

The oligo(beta-peptide)s, hexa(alpha-isobutyl beta,L-aspartate) (Hex-AIBLA) and octa(alpha-isobutyl beta,L-aspartate) (Oct-AIBLA), were synthesized in solution by using standard coupling methods. Powder x-ray diffraction showed that the octamer crystallized in the two helical crystal forms known to exist in the homologous poly(beta-peptide), whereas the hexamer seemed to adopt an extended conformation. Both CD and 1H-NMR spectra of Oct-AIBLA in MeOH revealed the presence of a regularly folded conformation in this solvent, presumably the 3(14) helix. The helix-to-coil transition of Oct-AIBLA was observed to take place upon heating in both MeOH and CHCl3, in the second case associated with a not-well-defined aggregation-disaggregation process. The spectroscopic evidence obtained on the presence of folded structures in Hex-AIBLA were much weaker than for the octamer.

Circular Dichroism↗

[Development of an experimental model for the study of the effects of cryotherapy on lung tumours].

UNLABELLED: Adenocarcinomas are today the most frequent lung cancers. They are mainly treated by surgery or by chemotherapy, but for the most advanced stages a local cryotherapy can be proposed as a palliative option for bronchial desobstruction. AIM OF THE STUDY: The aim of this work was to establish an experimental model to study in vivo the biological effects of this technique to propose it as a neoadjuvant treatment. MATERIALS AND METHODS: A xenograft system was used: cells from the A549 cell line were injected subcutaneously into SCID mice. Tumour nodes could be treated after seven weeks. The histological study showed that these tumours faithfully reproduced the morphological features of adenocarcinoma, and developed an intratumoral neovascularization. Two protocols of cryotherapy (1 vs 3 cycles of freezing) were performed and the induction of apoptosis was analyzed by immunohistochemical staining of cleaved caspase-3. RESULTS: The basal expression of cleaved caspase-3 in untreated tumours (23%) increased after cryotherapy. The increase was maximal eight hours after treatment (up to 47% of positive cells) and was less important with the first protocol, suggesting a lesser efficiency in the induction of apoptosis. CONCLUSION: The establishment of this model, which is faithful to physiological features, allowed us to demonstrate in vivo time and dose-dependent effects of cryotherapy.

Adenocarcinoma↗

Immunological classification of acute myeloblastic leukemias: relevance to patient outcome.

Immunophenotyping is a major tool to assign acute leukemia blast cells to the myeloid lineage. However, because of the large heterogeneity of myeloid-related lineages, no clinically relevant immunological classification of acute myeloblastic leukemia (AML) has been devised so far. To attempt at formulating such a classification, we analyzed the pattern of expression of selected antigens, on blast cells collected at AML diagnosis. Patients were eligible if they had a first diagnosis of de novo AML and a sufficient number of blast cells for proper immunophenotyping. The relative expression of CD7, CD13, CD14, CD15, CD33, CD34, CD35, CD36, CD65, CD117, and HLA-DR were analyzed by cytometry in a test series of 176 consecutive AML cases. Statistical tools of clusterization allowed to remove antigens with overlapping distribution, leading us to propose an AML classification that was validated in a second AML cohort of 733 patients. We identified five AML subsets (MA to ME) based on the expression of seven antigens within four groups (CD13/CD33/CD117, CD7, CD35/CD36, CD15).-MA and MB-AML have exclusively myeloid features with seldom extramedullary disease and rare expression of lymphoid antigens. No cases of acute promyelocytic leukemia (APL) were observed within MB AML. MC AML have either myeloid or erythroblastic features. MD AML have more frequently high WBC counts than other subsets, which were related to the expression of CD35/CD36 and CD14 and to monoblastic differentiation. ME AML lack CD13, CD33, and CD117 but display signs of terminal myeloid differentiation. Specific independent prognostic factors were related to poor overall survival in each immunological subset: CD34+ (P<3 x 10(-4)) in MA AML, CD7+ in MB AML, non-APL cases (P<0.03) in MC AML, CD34+ (P<0.002) and CD14+ (P<0.03) in MD AML, CD14+ in ME AML (P<0.01). The inclusion of seven key markers in the immunophenotyping of AML allows a stratification into clinically relevant subsets with individual prognostic factors, which should be considered to define high-risk AML populations.

Adolescent↗

Poor anticoagulant response to tissue factor pathway inhibitor in patients with venous thrombosis.

Tissue factor pathway inhibitor (TFPI) is of major importance in regulating the coagulation triggering effects of tissue factor. An association between TFPI deficiency and thrombosis has still not been clearly demonstrated. We evaluated the anticoagulant activity of exogenous TFPI added either to the plasma of patients with venous thrombosis (n = 118) or to the plasma of healthy controls similar in terms of mean age and sex ratio (n = 107). A poor anticoagulant response to TFPI, defined as TFPI resistance, was observed in 4.7% of controls and in 11.0% of patients. TFPI resistance was associated with an almost threefold increase in the risk of thrombosis and could therefore represent a novel hemostatic risk factor for venous thrombosis.

Adult↗

Mechanisms of staurosporine induced apoptosis in a human corneal endothelial cell line.

BACKGROUND: Apoptosis very probably plays a key part in endothelial cell loss during corneal storage in organ culture as well as hypothermic storage. However, the mechanisms underlying endothelial apoptosis are poorly understood. The response of a human corneal endothelial cell (HCEC) line to staurosporine, a known inducer of apoptosis, was investigated to gain insights into the intracellular modulators that participate in endothelial cell death. METHODS: Immortalised HCECs were studied after 3, 6, 12, and 24 hours of incubation with 0.2 micro M staurosporine. Cell shedding was monitored. Hoechst 33342 fluorescent DNA staining combined with propidium iodide was used for apoptosis/necrosis quantification and morphological examination. The caspase-3 active form was assessed using western blot, proteolytic activity detection, and immunocytochemistry. The cleaved form of poly(ADP-ribose) polymerase (PARP) was assessed using immunocytochemistry and western blot. The ultrastructural features of cells were screened after 12 hours with staurosporine or vehicle. RESULTS: The specific apoptotic nature of staurosporine induced HCEC death was confirmed. The ultrastructural features of staurosporine treated cells were typical of apoptosis. HCEC shedding and DNA condensation increased with time. Caspase-3 activity was detected as early as 3 hours after exposure with staurosporine, peaking at 12 hours of incubation. The presence of cleaved PARP after 3 hours confirmed caspase-3 activation. CONCLUSIONS: These data suggest strongly that HCEC cell death induced by staurosporine is apoptosis. The main consequence of HCEC apoptosis is shedding. Staurosporine induced apoptosis of endothelial cells involves activation of caspase-3, and could be a useful model to study strategies of cell death inhibition.

Apoptosis↗

Investigation of maximum doses absorbed by people accompanying patients in nuclear medicine departments.

In nuclear medicine, radioisotopes are bound to several compounds (called radiopharmaceuticals) for use in diagnostic and therapeutic applications. These unscaled sources are administered in various forms to patients, who remain radioactive for hours or days, and represent a source of potential radiation exposure for others. Thus, in nuclear medicine departments, radiation protection of workers and members of the public must take this exposure into account. In this context, the aim of this work was the investigation of maximum dose levels to which people accompanying patients are exposed. For monitoring, thermoluminescence dosemeters were given to 380 people in three nuclear medicine departments in Brazil. Exposure results are given, and issues regarding length of stay of members of the public in those departments are discussed.

Absorption↗

Value of two mortality assessment techniques for organ cultured corneal endothelium: trypan blue versus TUNEL technique.

BACKGROUND/AIM: It is known that trypan blue staining is not a good predictor of loss of corneal endothelial cells (ECs) during organ culture. As it is primarily an indicator of membrane integrity, it would also not be expected to identify ECs undergoing apoptosis. The aim of this study was to determine the ability of the in situ TdT dUTP mediated nick end labelling (TUNEL) technique to detect cell death in the corneal endothelium caused by apoptosis during organ culture, compared with conventional vital staining with trypan blue. METHODS: 31 human corneas were organ cultured at 31C for 3-35 days. Staurosporine was used to induce apoptosis in five control corneas. The endothelium was assessed by trypan blue and by the in situ TUNEL technique. The percentages of trypan and TUNEL positive ECs were compared. Their links with sex, donor age, time from donor death and organ culture, initial and final EC density and cell loss were studied. RESULTS: TUNEL stained ECs were observed in all corneas. TUNEL positive ECs were mostly located either in corneal folds or at the periphery of corneal folds showing central shedding. The mean percentage of cell death at the end of storage, assessed by the trypan blue technique, was 1.47% (SD 2.63, range 0.03-12); assessed by the TUNEL technique it was 12.7% (SD 16.4 range 0.6-65.5). There was a significant correlation between the two techniques (r = 0.7, p<0.001). The percentage of TUNEL stained ECs was correlated negatively with EC density at the end of storage (r = -0.47, p <0.005) and positively with percentage EC loss during storage (r = 0.46, p < 0.05). CONCLUSION: This study demonstrates that organ cultured corneas systematically carry non-viable ECs that are implicated in cell death by apoptosis and go undetected when trypan blue staining is used. Because the in situ TUNEL assay detects earlier events in the cell death process than does trypan blue, it should be used to quantify endothelial viability, especially for experiments with new storage media.

Adult↗

Automated tri-image analysis of stored corneal endothelium.

BACKGROUND: Endothelial examination of organ culture stored corneas is usually done manually and on several mosaic zones. Some banks use an image analyser that takes account of only one zone. This method is restricted by image quality, and may be inaccurate if endothelial cell density (ECD) within the mosaic is not homogeneous. The authors have developed an analyser that has tools for automatic error detection and correction, and can measure ECD and perform morphometry on multiple zones of three images of the endothelial mosaic. METHODS: 60 human corneas were divided into two equal groups: group 1 with homogeneous mosaics, group 2 with heterogeneous ones. Three standard microscopy video images of the endothelium, graded by quality, were analysed either in isolation (so called mono-image analysis) or simultaneously (so called tri-image analysis), with 50 or 300 endothelial cells (ECs) counted. The automated analysis was compared with the manual analysis, which concerned 10 non-adjacent zones and about 300 cells. For each analysis method, failures and durations were studied according to image quality. RESULTS: All corneas were able to undergo analysis, in about 2 or 7.5 minutes for 50 and 300 ECs respectively. The tri-image analysis did not increase analysis time and never failed, even with mediocre images. The tri-image analysis of 300 ECs was always most highly correlated with the manual count, particularly in the heterogeneous cornea group (r=0.94, p<0.001) and prevented serious count errors. CONCLUSIONS: This analyser allows reliable and rapid analysis of ECD, even for heterogeneous endothelia mosaics and mediocre images.

Aged↗

[Automated analyser of organ cultured corneal endothelial mosaic].

PURPOSE: Until now, organ-cultured corneal endothelial mosaic has been assessed in France by cell counting using a calibrated graticule, or by drawing cells on a computerized image. The former method is unsatisfactory because it is characterized by a lack of objective evaluation of the cell surface and hexagonality and it requires an experienced technician. The latter method is time-consuming and requires careful attention. We aimed to make an efficient, fast and easy to use, automated digital analyzer of video images of the corneal endothelium. METHODS: The hardware included a PC Pentium III ((R)) 800 MHz-Ram 256, a Data Translation 3155 acquisition card, a Sony SC 75 CE CCD camera, and a 22-inch screen. Special functions for automated cell boundary determination consisted of Plug-in programs included in the ImageTool software. Calibration was performed using a calibrated micrometer. Cell densities of 40 organ-cultured corneas measured by both manual and automated counting were compared using parametric tests (Student's t test for paired variables and the Pearson correlation coefficient). RESULTS: All steps were considered more ergonomic i.e., endothelial image capture, image selection, thresholding of multiple areas of interest, automated cell count, automated detection of errors in cell boundary drawing, presentation of the results in an HTML file including the number of counted cells, cell density, coefficient of variation of cell area, cell surface histogram and cell hexagonality. The device was efficient because the global process lasted on average 7 minutes and did not require an experienced technician. The correlation between cell densities obtained with both methods was high (r=+0.84, p<0.001). The results showed an under-estimation using manual counting (2191+/-322 vs. 2273+/-457 cell/mm(2), p=0.046), compared with the automated method. CONCLUSIONS: Our automated endothelial cell analyzer is efficient and gives reliable results quickly and easily. A multicentric validation would allow us to standardize cell counts among cornea banks in our country.

Aged↗

Practice of nuclear medicine in Spain.

This survey presents the results of a poll sent to all Spanish nuclear medicine departments between July 1999 and March 2000, with the aim of clarifying the current situation of nuclear medicine in Spain. This survey is believed to be the first of its kind, and it is anticipated that the data will be of assistance to health authorities in ensuring that the needs of the population with regard to nuclear medicine facilities are met.

Nuclear Medicine↗

[Tomographic studies of myocardial perfusion normality with 99m Tc-Tetrofosmin. Spanish multicenter study].

As a first phase in a nationwide multicenter study to obtain myocardial perfusion normality patterns, this work presents the study design and quality control methodology used to guarantee that the gammacameras fulfilled some minimum quality requirements. The following aspects were considered in the study design in order to guarantee the homogeneity and interchangeability of the results: creation and structure of the work group, data interchange system, data selection and acquisition, centralized archiving and processing, assessment system, study acceptance criteria and distribution of the results. To carry out the instrumental quality control, three phantom studies were established, one to control the rotation center, another to verify tomographic uniformity and a third to simulate the shape and orientation of the left ventricle. The three phantoms circulated through all of the 18 participating centers in this project, which corresponded to 19 gammacameras. Very strict guidelines had to be followed in the acquisition and processing of these phantom studies. If any camera and/or center did not fulfill the criteria established, it was advised of the problem detected in order to correct it. Once the defect was repaired, all the phantoms were sent again for verification. Uniformity of the rotation center was quantified by means of the eccentricity of a 360 degrees orbit, admitting up to a maximum of 10%. Tomographic uniformity was visually assessed, taking in account the number of slices with rings and their contrast and finally no artifacts could be present in the reconstructed study of the ventricle. The center of rotation was within limits in all the cameras except one case while the tomographic uniformity was incorrect in 6 cases. All the departments, except one, corrected the defects detected, and passed the acceptance test. The results made it possible to guarantee adequate homogeneity and instrumental quality in this multicenter study.

Coronary Circulation↗

[Tomographic studies of myocardial perfusion normality with 99m Tc-Tetrofosmin. Spanish multicenter study].

This objective of this study was to obtain a pool of Myocardial Perfusion SPECTs with a 99mTetrofosmin stress/rest one day protocol in normal volunteers having a less than or equal to 5% likelihood of coronary artery disease that represents normalcy in the Spanish population. A total of 169 volunteers from 15 hospitals were studied. The volunteers were divided into 5 groups: Groups 1, 2 or 3 corresponding to men < 30 years (n = 33), men between 30 and 50 years (n = 32), or men > 50 years (n = 31); Groups 4 or 5: premenopausal (n = 38) or postmenopausal women (n = 35). A clinical history, physical examination, clinical laboratory parameters, echocardiography and a symptom limited exercise stress test were performed in all of them and had to be normal. The mean likelihood of coronary artery disease was 1.15 +/- 1.07%.Twenty-four segments were analyzed in each study and were classified into 5 grades of uptake (1 = normal, 2, 3, 4 = mild, moderate or severe defect and 5 = no uptake). Defects were then analyzed according to sex and location. Considering the stress and rest studies separately (8,112 segments), only 19 moderate and 75 mild defects were found, these corresponding to 16 volunteers, with more inferior defects in men and anterior defects in women. These data validate the normalcy of our population. A pool of Myocardial Perfusion SPECTs with a 99mTetrofosmin stress/rest one day protocol in normal volunteers that represents Spanish normal values was obtained.

Adult↗