A multicenter, randomized, double-blind, placebo-controlled, dose-finding trial of a long-acting formulation of octreotide in promoting weight loss in obese adults with insulin hypersecretion.

OBJECTIVE: To compare changes in weight in obese patients who received long-acting octreotide (octreotide LAR) at one of three dose levels (20, 40, or 60 mg) or placebo over 6 months and to identify the lowest dose of octreotide LAR that safely achieved optimal weight loss. DESIGN: Randomized, double-blind, placebo-controlled trial of octreotide LAR at three dose levels. PATIENTS: A total of 172 adults (28 men and 144 women) with at least moderate obesity (body mass index (BMI) range 30-65 kg/m2) and evidence of insulin hypersecretion were enrolled. Patients were predominantly either Caucasian (50.0%) or African American (45.3%). The mean age (38 +/- 11 year), weight (110.7 +/- 23 kg), and BMI (39.8 +/- 6.5 kg/m2) were similar across the four treatment groups. MEASUREMENTS: Efficacy measures included weight, BMI, fasting serum glucose; triglycerides; percentage of total body fat and abdominal fat as measured by dual-energy X-ray absorptiometry; skin fold thickness; waist-to-hip circumference; leptin; percentage of carbohydrates, fat, and protein ingested; nutritional evaluation (including dietary analysis--3-day food record); quality of life (QoL; using the Impact of Weight on Quality of Life-Lite); Beck Depression Inventory; and Carbohydrate Craving Questionnaire. Safety measures included medical history, vital signs, physical examinations, hematology, blood chemistries, thyroid function tests, hemoglobin A1c, gallbladder ultrasound, electrocardiograms, and adverse events. RESULTS: After 6 months of treatment, patients receiving 40 or 60 mg of octreotide LAR experienced statistically significant weight loss compared to baseline, with mean differences from placebo in percent weight change of -1.98 and -1.87%, respectively. This finding was accompanied by statistically significant mean decreases in BMI compared to baseline, that is, a mean decrease of 0.73 and 0.79 kg/m2 for the 40 and 60 mg treatment arms, respectively. The observed weight loss was progressive during the 6-month treatment in the two higher dose groups. The lowest dose to reach statistical significance in weight loss after 6 months' treatment was 40 mg. Post hoc analysis revealed a 3.5-3.8% weight loss at month 6 in the two higher dose groups among Caucasian patients having insulin secretion greater than the median of the cohort, defined as CIR(gp) (corrected insulin response at the glucose peak) > or = 1.43. There were no statistically significant changes in QoL scores, body fat, leptin concentration, Beck Depression Inventory, or macronutrient intake. Mean changes of blood glucose AUC(0-180 min) during an oral glucose tolerance test in patients taking octreotide LAR were 39-40 mg/dl h higher than those on placebo. A total of 7-21% of the patients taking octreotide LAR reached a 5% or greater decrease in body weight from Baseline, compared to 11% for the placebo group. This was not statistically significant. The most common adverse events included diarrhea, headache, cholelithiasis, nausea, and abdominal pain. CONCLUSION: Octreotide LAR given at 40 or 60 mg resulted in statistically significant weight loss. A post hoc analysis stratifying patients by race and CIR(gp) indicated that Caucasian patients with the greater degree of insulin hypersecretion appeared to derive the most benefit from treatment. The observed safety profile was consistent with the known effects of octreotide from previous studies.

Tipos histológicos y variables pronósticas en 569 casos de melanoma maligno de la piel, entre 1976 y 2001 / Histological types in 569 cases of malignant melanoma of the skin, between 1976 and 2001

El melanoma maligno (MM) primario de la piel ha incrementado su incidencia en todos los países del mundo, afectando a personas de todas las edades y de ambos sexos. Para conocer la frecuencia de tipos histológicos y de las variables pronósticas del melanoma maligno primario de la piel en nuestro hospital, se revisaron las biopsias del Departamento de Anatomía Patológica entre enero de 1976 y diciembre de 2001. Se revisaron 238.547 biopsias entre 1976 y 2001, encontrándose 569 casos de MM, 305 mujeres y 255 hombres; en 9 casos no se consignó el sexo. El promedio de edad de los casos fue de 51,8 años, sin diferencias entre hombres y mujeres. Del total de casos, 429 eran MM invasor (75,4 por ciento) y 140 MM in situ (24,6 por ciento). El tipo histológico del MM invasor más frecuentemente encontrado fue melanoma de tipo extensión superficial (27,4 por ciento), seguido por melanoma tipo nodular (17 por ciento). El promedio de espesor según Breslow para MM invasor fue de 1,2 mm. El MM de la piel es una neoplasia cada vez más frecuente en nuestro medio. En nuestra casuística tiende a predominar en mujeres y afecta a adultos en la edad media de la vida. Un porcentaje significativo de lesiones (24,6 por ciento) se detecta en una fase in situ, lo que es comparable al de estudios poblacionales norteamericanos.

The influence of speed, grade and mass during simulated off road bicycling.

The purpose of this investigation was to examine the effects of bicycle mass, speed, and grade on oxygen consumption (VO2), heart rate (HR), and ratings of perceived exertion (RPE) during a simulated off-road riding paradigm. Nine adult subjects with mean +/- SD age, mass, and VO2 max of 26.1 +/- 5.6 years, 71.7 +/- 7.5 kg, 56.6 +/- 5.2 ml x kg(-1) x min(-1) respectively, were trained to ride a fully suspended Trek Y-22 mountain bike on a treadmill with a 3.8 cm bump affixed to the belt. Riders completed a maximum of nine separate trials encompassing three different bike masses (11.6, 12.6 and 13.6 kg), 3 speeds (2.7, 3.6 and 4.5 m x s(-1)), and 3 grades (0, 2.5, and 5%). Throughout a trial, bike mass and speed remained constant while riding grade was increased every 5 min. During simulated off-road riding on a fully suspended mountain bike, increases in speed and grade significantly increased VO2, heart rate, and RPE. Increases in bike mass had no significant effects on VO2, heart rate or RPE. In addition, speed and grade changes interacted to differentially affect VO2, heart rate, and RPE at all speeds and grades.

[Sweet syndrome associated with articular and renal involvement]. / Síndrome de Sweet asociado a compromiso articular y renal.

We report a 29 years old female presenting with fever and painful infiltrated erythematous and violaceous plates with pseudo vesicles in the surface, located in both arms, four days after having suffered a tonsillitis. She was admitted with the diagnosis of Sweet syndrome and the lesions disappeared spontaneously. Two months later, she presented with a similar condition, again after an upper respiratory infection. Five months later, she was admitted with arthralgias with positive rheumatoid factor and antinuclear antibodies. Three years after the first admission, she was admitted with an acute glomerulonephritis and renal failure after another upper respiratory infection. Sweet syndrome was described in 1964 and, although initially considered benign, its association with inflammatory diseases or cancer has been reported.

Wheat DNA polymerase CI: a homologue of rat DNA polymerase beta.

We have previously described a low-molecular-weight DNA polymerase (52 kDa) from wheat embryo: DNA polymerase CI (pol CI). This enzyme shares some biochemical properties with animal DNA polymerase beta (pol beta). In this report, we analyse pol CI in wheat embryo germination. Immunodetection and measurement of the enzyme activity show that wheat pol CI remains at a constant level during germination, whereas dramatic changes of the replicative DNA polymerase A and B activities were previously reported. We observe that the level of pol CI in physiological conditions (embryo germination and dividing cell culture) is in agreement with a pol beta-type DNA polymerase. By microsequencing of the electroblotted 52 kDa polypeptide, we determined the sequence of a dodecapeptide from the N-terminal region. A comparative analysis of the N-terminal pol CI peptide with some mammalian pol beta sequences shows a clear homology with helix 1 of the N-terminal ssDNA domain (residues 15 to 26) of the rat pol beta. Thus, the helical structure of this region should be conserved in the wheat peptide. This represents the first evidence of a partial primary structure of a beta-type DNA polymerase in plants.

Ferroelectric Field Effect Transistor Based on Epitaxial Perovskite Heterostructures

Ferroelectric field effect devices offer the possibility of nonvolatile active memory elements. Doped rare-earth manganates, which are usually associated with colossal magnetoresistive properties, have been used as the semiconductor channel material of a prototypical epitaxial field effect device. The carrier concentration of the semiconductor channel can be "tuned" by varying the manganate stochiometry. A device with La0.7Ca0.3MnO3 as the semiconductor and PbZr0.2Ti0.8O3 as the ferroelectric gate exhibited a modulation in channel conductance of at least a factor of 3 and a retention loss of 3 percent after 45 minutes without power.

Changes of enzymes and factors involved in DNA synthesis during wheat embryo germination.

We have previously purified and characterized wheat germ DNA polymerases A and B. To determine the role played by DNA polymerases A and B in DNA replication, we have measured the level of their activities during wheat embryo germination. The level of cellular proteins known to be associated with DNA synthesis such as PCNA and DNA primase were also investigated. The activity of DNA polymerase A gradually increased reaching a maximal level at 12 h after germination. Three days later, only a residual activity was detected. DNA polymerase B showed the same pattern during germination with very similar changes in activity. Our results indicate a striking correlation between maximal activities of DNA polymerase A, DNA polymerase B and optimal levels of DNA synthesis. These results support a replicative role of these enzymes. The activity of wheat DNA primase that copurifies with DNA polymerase A also increases during wheat germination. Taking together all its properties, and in spite of its behaviour with some inhibitors. DNA polymerase A may be considered as the plant counterpart of animal DNA polymerase alpha. Concerning DNA polymerase B we have previously shown that PCNA stimulates its processivity. Besides studying the changes of DNA polymerases A and B and DNA primase we have also studied changes in PCNA during germination. We show that PCNA is present in wheat embryos at a constant relatively high level during the first 24 h of germination. After 48 h, the absence of PCNA is concomitant with an important decrease in DNA polymerase B activity. In this report we confirm the behaviour of DNA polymerase B as a delta-like activity.

A Philadelphia chromosome positive CML patient with a unique translocation studied via GTG-banding and fluorescence in situ hybridization.

The Philadelphia (Ph) chromosome was the first consistently occurring chromosome abnormality associated with a single cancer type, chronic myelogenous leukemia (CML). This translocation has since been reported with other chromosome abnormalities. The present report describes a case of Ph chromosome positive CML with a unique complex translocation identified using molecular cytogenetics in addition to routine techniques. GTG-banding revealed abnormalities in at least chromosomes 9, 13, 17, and 22. Fluorescence in situ hybridization (FISH) studies were performed as an adjunct to conventional cytogenetic analyses. Using FISH with the Oncor bcr/abl probe, the Ph translocation previously hypothesized was confirmed. Applying FISH with paired painting probes in various combinations, a complex translocation involving chromosomes 9, 13, 15, 17, and 22 was observed. The results of the GTG-banding and FISH studies were compared with each other and correlated with those of the hematological findings. In an extensive search of the medical literature database (Medline, Health, Cancerlit, Ovid, and CINAHL) spanning nearly three decades (1965-1994), we found no previous report of this specific translocation. Therefore, to the best of our knowledge, this is a unique translocation associated with Ph chromosome positive CML.

Quantitation of mRNA species by RT-PCR on total mRNA population.

PCR is commonly used for mRNA quantitation. Previously described procedures are applied to one or a few specific mRNA sequences. We show here that methods used for amplifying heterogeneous cDNA populations can be applied to the quantitation of many mRNA species. This quantitation is achieved by dot blotting and hybridization with the corresponding probes after amplifying a bulk mRNA population. Only a single, two-round-amplification assay is required for quantitation of a whole set of mRNA species. The proportionality of input molecules to output signal was shown by performing a series of control experiments. We applied this technique to measure the relative variations of the MBP, Po, and MAG mRNA sequences in the normal trembler mouse model. The results were consistent with previously described Northern blot data. This quantitative PCR method provides a rapid and reliable way to quantify relative amounts of mRNA species in small amounts of total RNA by using internal controls.

Gamma interferon potently induces tryptophanyl-tRNA synthetase expression in human keratinocytes.

Incubation of human keratinocytes with gamma interferon (gamma-IFN) induces the synthesis of a 53-kDa protein of unknown nature and function. We report the identification of this protein through amino acid microsequencing. The NH2-terminal amino acid sequence of the 53-kDa antigen demonstrated that this protein was tryptophanyl-tRNA synthetase (Frolova et al, Gene 109:291-296, 1991, Genbank accession number 61715). This result was validated by the sequencing of tryptic peptides. Identification of the 53-kDa gamma-IFN-induced protein was confirmed by immunoblotting with an antiserum directed against beef pancreas tryptophanyl-tRNA synthetase. Northern blot analysis using a synthetic oligonucleotidic 32P-labeled probe evidenced a 3.1-kb transcript in gamma-IFN-treated cells indicating that the gene was regulated at the pre-translational level. These data show that gamma-IFN potently induces in keratinocytes the expression of an enzyme directly involved in protein biosynthesis. Elevated levels of tryptophanyl-tRNA synthetase in treated cultured keratinocytes might be involved in the cell-growth-inhibitory activity of gamma interferon.

Accuracy of fine-needle aspiration biopsy in diagnosis of palpable breast masses: two-year results of a university-affiliated community hospital.

Fifty-eight solid breast masses in 57 patients were subjected to fine-needle aspiration biopsy, and the results of cytologic analysis and histologic diagnosis were compared. The sensitivity was 79.2%, specificity, 85.3%, and accuracy, 82.8%. These results are within the range of those in the medical literature. Fine-needle aspiration biopsy is a useful adjunct in the management of palpable breast masses in the community hospital setting. Our data, however, do not support its use alone as a screening tool. Excisional biopsy or close clinical follow-up remains essential for evaluation and management of palpable breast masses.

A mammalian tryptophanyl-tRNA synthetase shows little homology to prokaryotic synthetases but near identity with mammalian peptide chain release factor.

Determination of the amino acid sequence of beef pancreas tryptophanyl-tRNA synthetase was undertaken through both cDNA and direct peptide sequencing. A full-length cDNA clone containing a 475 amino acid open reading frame was obtained. The molecular mass of the corresponding peptide chain, 53,728 Da, was in agreement with that of beef tryptophanyl-tRNA synthetase, as determined by physicochemical methods (54 kDa). Expression of this clone in Escherichia coli led to tryptophanyl-tRNA synthetase activity in cell extracts. The open reading frame included two sequences analogous to the consensus sequences, HIGH and KMSKS, found in class I aminoacyl-tRNA synthetases. The homology with prokaryotic and yeast mitochondrial tryptophanyl-tRNA synthetases was low and was limited to the regions of the consensus sequences. However, a 90% homology was observed with the recently described rabbit peptide chain release factor (eRF) [Lee et al. (1990) Proc. Natl. Acad. Sci. 87, 3508-3512]. Such a strong homology may reveal a new group of genes deriving from a common ancestor, the products of which could be involved in tRNA aminoacylation (tryptophanyl-tRNA synthetase) or translation termination (eRF).

Transhiatal blunt esophagectomy for carcinoma of the esophagus.

Transhiatal blunt esophagectomy has been reported as a safe and effective procedure for the palliation of carcinoma of the esophagus. Avoidance of a thoracotomy eliminates the morbidity associated with this procedure, and creation of a cervical esophagogastric anastomosis avoids the catastrophic sequelae of an intrathoracic anastomotic leak. Moreover, use of the procedure for palliation does not preclude excellent 1-year survival rates. We report early results in five consecutive patients with esophageal carcinoma who underwent transhiatal blunt esophagectomy. Five patients had 22 complications, including one with a fascial dehiscence, pyloroplasty leak, and localized mediastinal abscess requiring a second laparotomy. One patient died in the hospital postoperatively of massive aspiration pneumonitis. Our results compare favorably with those reported in the literature. We believe that transhiatal blunt esophagectomy avoids the morbidity and mortality of a thoracotomy and an intrathoracic anastomosis, yet remains a major gastrointestinal operative procedure with all of its attendant risks.

Tryptophanyl-tRNA synthetase-like immunoreactivity in the central nervous system and midgut of the migratory locust. Comparisons with gastrin-cholecystokinin-like and octopamine-like immunoreactivity.

A tryptophanyl-tRNA synthetase (TrpRS)-immunoreactivity is localized in various neurosecretory cells of all ganglia of the central nervous system of the Orthoptera Locusta migratoria, except in deutocerebrum, and in endocrine cells of the midgut. It has been observed that TrpRS-like material never co-localizes either with CCK-like or octopamine-like material. TrpRS immunoreactive perikarya and processes that ramify extensively throughout the neuropiles have been detected in the protocerebrum, optic lobes, tritocerebrum, suboesophageal, thoracic and abdominal ganglia. In the lateral protocerebrum, a particular TrpRS pathway different from the lateral gastrin cholecystokinin (CCK-8(s] pathway is revealed, certain of these processes terminating in the glandular part of the corpora cardiaca. In the metathoracic ganglion, have been observed numerous immunoreactive cell bodies and processes in the neuropiles. Some of them constitute a major pathway and which are distinct from octopamine (OA) cells but in close vicinity with the latter. In the midgut immunopositive TrpRS-like cells are dispersed among the regenerative and digestive cells of the epithelium; they are different from gastrin-cholecystokinin positive cells. The various TrpRS-like immunoreactivities identified in Locusta indicate that TrpRS-like material may occur in different tissues of organisms other than Vertebrates. These results suggest also that TrpRS-like enzyme could be involved in functions other than aminoacylation, as in Vertebrates.

Tryptophanyl-tRNA synthetase is a major soluble protein species in bovine pancreas.

Besides their central role in protein synthesis, aminoacyl-tRNA synthetases have been found or thought to be involved in other processes. We present here a study showing that tryptophanyl-tRNA synthetase has a surprising tissular distribution. Indeed, immunochemical determinations showed that in several bovine organs such as liver, kidney and heart, tryptophanyl-tRNA synthetase constitutes, as expected, about 0.02% of soluble proteins. In spleen, brain cortex, stomach, cerebellum or duodenum, this amount is about 10-times higher, and in pancreas it is 100-fold. There is no correlation between these amounts and the RNA content of the organs. Moreover, the concentration of another aminoacyl-tRNA synthetase (methionyl-tRNA synthetase) is higher in liver than in pancreas, while the amount of tRNATrp is not higher in pancreas than in liver as compared to other tRNAs. Among several interpretations, it is possible that tryptophanyl-tRNA synthetase is involved in a function other than tRNA aminoacylation. This unknown function would be specific to the differentiated organs, since fetal cerebellum and fetal pancreas contain the same amount of tryptophanyl-tRNA synthetase as adult liver.

Characterization of a murine monoclonal antibody specific for an allotypic determinant on T cell antigen receptor.

Repeated immunization of normal C57L/J (H-2b) mice with peripheral T cells from BALB.B (H-2b) mice results in the production of antibodies which react with the T cell receptor. A monoclonal antibody-producing hybridoma, F23.1, was isolated from immunized C57L/J mice showing this property. This monoclonal antibody recognizes approximately 25% of peripheral T cells in BALB mice. It stains approximately the same fraction of T cells and precipitates the same heterodimer as the rat monoclonal antibody described previously that was made against isolated receptor material. The allotypic determinant recognized by this monoclonal antibody is present in most common laboratory strains (BALB, C57BL, CBA, A, DBA, C3H) and is absent in C57L, C57BR, and SJL mice. Sorting peripheral T cells from BALB.B or (SJL X BALB/c)F1 mice for the F23.1+ and F23.1- subsets revealed that both populations contain approximately the same CTL precursor frequency for alloantigen. Thus, the T cell receptor allotype defined by F23.1 is present on CTL. Furthermore, cytotoxicity mediated by an F23.1+ CTL line could be blocked specifically by the F23.1 monoclonal antibody. Under appropriate conditions, the monoclonal antibody F23.1 bound to Sepharose 4B beads can induce resting peripheral T lymphocytes of allotype-positive strains to proliferate.

Studies on the mechanism of suppression of primary cytotoxic responses by cloned cytotoxic T lymphocytes.

We have shown in the accompanying companion paper that cloned cytotoxic T lymphocytes (CTL) can serve as veto cells in vitro, suppressing primary cytotoxic activity directed against antigens expressed by those cloned CTL but not against third party antigens. We now explore the mechanism of this antigen-specific suppression by cloned CTL, using as a model system the ability of G4, a BALB.B anti-H-2Dd CTL clone, to specifically suppress a primary in vitro anti-H-2b CTL response. G4 cells do not constitutively secrete a suppressor factor, because suppression cannot be mediated by supernatants removed from G4 cells at a time when they are routinely used as veto cells. Furthermore, medium removed from cultures suppressed by G4 will not suppress, indicating that the veto cell function of G4 is not mediated by soluble factors. Full suppression of primary anti-H-2b CTL responses requires that G4 be present throughout the 5-day mixed lymphocyte culture (MLC). Removal of G4 during the first 3 days of MLC results in a drastic reduction in the level of antigen-specific suppression, with a slight but reproducible loss of suppression after veto cell removal on day 4. The addition of G4 during the course of an ongoing MLC reveals that maximal suppression requires the presence of veto cells during the first 24 to 48 hr of culture. Thus, G4 cells must be present both early and late in an MLC to exert maximal veto cell suppression. Several experiments suggest that G4-induced veto cell activity is unlikely to be due to cytolysis of CTL precursors which are capable of recognizing G4. G4 cannot specifically recognize these CTL precursors, and G4 cells are inefficient at lectin-mediated lysis of non-tumor cell targets. Furthermore, we show that G4 cells cannot lyse CTL which recognize them. Finally, dilutions of anti-clonotypic antibodies which completely block both lectin-mediated and specific cytolysis by G4 do not block (and in fact enhance) G4-mediated veto cell activity.

Monoclonal antibodies specific for a murine cytotoxic T-lymphocyte clone.

Two antibody-secreting murine hybridomas, F1G3.1 and F2A11.5, have been established from B10.D2 mice immunized with cells from the murine cytotoxic T-lymphocyte clone G4. The two clones used, G4 and B10, were derived from BALB.B (H-2b) mice and the target antigen specificity of both maps to the Dd region of the murine H-2 complex. However, B10 has a lower affinity for the target cells, as shown by its lower specific killing of blasts and its higher susceptibility to blocking by anti-Lyt-2 monoclonal antibody 53-6.75. The monoclonal antibodies, F1G3.1 and F2A11.5, react only with cells from clone G4. Similarly, they block only the specific cytolysis mediated by G4; no effect on cytotoxicity mediated by B10 or by heterogeneous populations of cytotoxic T lymphocytes was found. F1G3.1, especially, is very active in stimulating G4 to secrete immune interferon; B10 in contrast did not show any induction on treatment with these monoclonal antibodies. The structure of the surface antigen on G4 cells recognized by these monoclonal antibodies was revealed by immunoprecipitation studies of radioiodinated cell surface proteins. A protein dimer could be identified with an apparent molecular size of 80,000 daltons consisting of monomers migrating as 42,000-dalton proteins on reduction. So far, electrophoresis in the presence of NaDodSO4 does not indicate any heterogeneity in the size of the monomers. This molecule can be distinguished from the Lyt-2 complex.

Role of thiol compounds in mammalian melanin pigmentation. II. Glutathione and related enzymatic activities.

Previously, we reported evidence suggesting that, in addition to tyrosinase, glutathione-reductase plays an important role in the regulation and control of the biosynthetic activity of melanocytes. Further investigations were performed on a mammal presenting a well-defined genotype for coat pigmentation, the mutant mouse [subline C57 BL (6J)], namely the nonagouti black (a/a) mutant and the yellow (Ay/a) mutant showing, respectively, pure uniform eumelanin and phaeomelanin pigmentation. Analysis of thiol compounds and glutathione-related enzyme levels in mouse skin gave similar results to those found in tortoise-shell guinea pig skin. The observed differences in the glutathione and glutathione-related enzyme content between black and yellow (or red) skin provide evidence that the increase of glutathione-reductase activity in the environment of the melanocytes may stimulate the pigment cells to produce phaeomelanin instead of eumelanin pigment.