Prevention of incisional hernia with a reinforced tension line (RTL) versus primary suture only in midline laparotomies: 3-year follow-up in a randomized clinical trial.

PURPOSE: Incisional hernia (IH) has an incidence of 10-23%, which can increase to 38% in specific risk groups. The objective of this study is to report the results at 3 years of follow-up of the use of the reinforced tension line (RTL) technique compared with primary suture only (PSO) closure in the prevention of IH in high-risk patients undergoing laparotomy. METHODS: Open randomized controlled clinical trial. Included were patients older than 18 years who underwent midline laparotomy, emergency or scheduled, who were considered high risk, and who completed 3-year follow-up. The patients were randomized 1:1 to the RTL technique or to PSO. The objective was to report the incidence of IH and the complications associated with the closure method. Intention-to-treat analysis and Cox regression were performed. RESULTS: A total of 124 patients were randomized; 51 patients from the RTL group and 53 patients from the PSO group finished the 3-year follow-up. The incidence of IH was higher in the PSO group (15/53, 28.3%) than the RTL group (5/51, 9.8%) (p = 0.016, OR 0.35, 95% CI 0.14-0.88, number needed to treat 5.4, log-rank test p = 0.017). The groups were similar in the rates of surgical site infection, hematoma, seroma, and postoperative pain during follow-up. CONCLUSIONS: The RTL technique is useful in the prevention of IH when compared with PSO in high-risk midline laparotomy patients, and it is not associated with a higher percentage of complications. TRIAL REGISTRATION: Local Committee CI-HRAEB-2013-020. March 13, 2013. CLINICAL TRIALS: NCT02136628, retrospectively registered.

Baseline 18F-Fluorocholine PET/CT and bone scan in the outcome prediction of patients treated with radium 223 dichloride.

AIM: To establish the utility of baseline 18F-Fluorocholine (FCH) PET/CT and bone scintigraphy (BS) in the outcome prediction of patients with castration-resistant prostate cancer and bone metastases (CRPC-BM) treated with 223Ra. METHODS: Prospective, multicenter and non-randomized study (ChoPET-Rad study). FCH PET/CT and BS were performed before the initiation of 223Ra (basal FCH PET/CT and BS). Bone disease was classified attending the number of lesions in baseline BS and PET/CT. FCH PET/CT was semiquantitatively evaluated. Gleason score, baseline levels of prostate-specific antigen (PSA), alkaline phosphatase and lactate dehydrogenase were determined. Progression-free survival (PFS) and overall survival (OS) since the onset of 223Ra treatment was calculated. PFS was defined by PSA rising. Relations between clinical and imaging variables with PFS and OS were evaluated by Pearson, Mann-Whitney tests and Kapplan-Meier analysis. Univariate and multivariate Cox regression analysis was performed. RESULTS: Forty patients were evaluated. The median PFS and OS were of 3.0 ± 2.3 and 23.0 ± 4.2 months, respectively. 33 patients progressed and 13 died during the follow-up. The extension of the bone disease by FCH PET/CT (p = 0.011, χ2 = 10.63), BS (p = 0.044, χ2 = 8.04), SUVmax (p = 0.012) and average SUVmax (p = 0.014) were related to OS. No significant association was found for the PFS. ROC analysis revealed significant association of SUVmax, average SUVmax and basal PSA with OS. Only therapeutic failure was associated with OS in the multivariate analysis (HR = 3.6, p = 0.04). CONCLUSION: FCH PET/CT and BS had prognostic aim in the prediction of OS. None clinical or imaging variable was able to predict the PFS, probably due to the high rate of progressive disease.

Cloning, characterization and functional expression of Taenia solium 17 beta-hydroxysteroid dehydrogenase.

The 17ß-hydroxysteroid dehydrogenases (17ß-HSD) are key enzymes involved in the formation (reduction) and inactivation (oxidation) of sex steroids. Several types have been found in vertebrates including fish, as well as in invertebrates like Caenorhabditis elegans, Ciona intestinalis and Haliotis diversicolor supertexta. To date limited information is available about this enzyme in parasites. We showed previously that Taenia solium cysticerci are able to synthesize sex steroid hormones in vitro when precursors are provided in the culture medium. Here, we identified a T. solium 17ß-HSD through in silico blast searches in the T. solium genome database. This coding sequence was amplified by RT-PCR and cloned into the pcDNA 3.1(+) expression vector. The full length cDNA contains 957bp, corresponding to an open reading frame coding for 319 aa. The highest identity (84%) at the protein level was found with the Echinococcus multilocularis 17ß-HSD although significant similarities were also found with other invertebrate and vertebrate 17ß-HSD sequences. The T. solium Tsol-17ßHSD belongs to the short-chain dehydrogenase/reductase (SDR) protein superfamily. HEK293T cells transiently transfected with Tsol17ß-HSD induced expression of Tsol17ß-HSD that transformed 3H-androstenedione into testosterone. In contrast, 3H-estrone was not significantly transformed into estradiol. In conclusion, T. solium cysticerci express a 17ß-HSD that catalyzes the androgen reduction. The enzyme belongs to the short chain dehydrogenases/reductase family and shares motifs and activity with the type 3 enzyme of some other species.

KCNH1 potassium channels are expressed in cervical cytologies from pregnant patients and are regulated by progesterone.

Potassium voltage-gated channel, subfamily H (eag-related), member 1 (KCNH1) potassium channels are potential tumour markers and cancer therapeutic targets and are up-regulated by oestrogens and human papilloma virus (HPV) oncogenes. However, the role of KCNH1 in normal tissues is poorly understood, and its expression in pregnancy is unknown. We wondered whether KCNH1 channels are expressed in cervical cells from pregnant patients and whether progesterone (P4) regulates KCNH1. The association with HPV was also investigated. KCNH1 protein expression was studied by immunocytochemistry in liquid-based cervical cytologies; 93 samples were obtained from pregnant patients at different trimesters, and 15 samples were obtained from non-pregnant women (controls). The presence of HPV was studied by PCR with direct sequencing and nested multiplex PCR. HeLa cervical cancer cells were transfected with human progesterone receptor-B (PR-B) and treated with P4. KCNH1 mRNA expression in these cultures was studied by real-time PCR. KCNH1 protein was detected in 100% of the pregnancy samples and in 26% of the controls. We found 18 pregnant patients infected with HPV and detected 14 types of HPV. There was no association between the percentage of cells expressing KCNH1 and either the presence or type of HPV. P4 induced KCNH1 mRNA and protein expression in cells transfected with human PR-B. No regulation of KCNH1 by P4 was observed in non-transfected cells. We show for the first time the expression of an ion channel during human pregnancy at different trimesters and KCNH1 regulation by P4 in human cells. These data raise a new research field for KCNH1 channels in human tissues.

Taenia crassiceps WFU cysticerci synthesize corticosteroids in vitro: metyrapone regulates the production.

Taenia solium and Taenia crassiceps WFU cysticerci and tapeworms have the ability to synthesize sex steroid hormones and have a functional 3ß-hydroxisteroid dehydrogenase. Corticosteroids (CS) like corticosterone and dexamethasone have been shown to stimulate in vitro estrogen production by Taenia crassiceps WFU cysticerci. The aim of this work was to study the ability of T. crassiceps WFU cysticerci to synthesize corticosteroids, and the effect of the inhibitor metyrapone on the CS synthesis. For this purpose T. crassiceps WFU cysticerci were obtained from the abdominal cavity of mice, thoroughly washed and pre-incubated in multiwells for 24 h in DMEM plus antibiotics/antimycotics. The tritiated CS precursor progesterone ((3)H-P4) was added to the culture media and parasites cultured for different periods. Blanks containing the culture media plus the (3)H-P4 were simultaneously incubated. Blanks and parasite culture media were ether extracted and analyzed by thin layer chromatography (TLC) in two different solvent systems. Corticosterone production was measured in the culture media by RIA. In some experiments metyrapone (0.1-0.5 mM) was added for 24, 48 or 72 h. Results showed that cysticerci mainly synthesized tritiated 11-deoxy corticosterone (DOC) and small amounts of corticosterone that was also detected by RIA. Small amounts of (3)H-11-deoxy cortisol were also found. Corticosteroid synthesis was time dependent. The addition of metyrapone significantly inhibited tritiated DOC, deoxycortisol and corticosterone synthesis. These results show for the first time that parasites have the capacity to synthesize CS that is modulated by metyrapone. Data suggest that DOC is the main corticosteroid in the parasites.

The effect of glucocorticoids on sex steroid synthesis in cultured Taenia crassiceps Wake Forest University (WFU) cysticerci.

We have shown previously that cultured Taenia crassiceps Wake Forest University (WFU) and Taenia solium cysticerci, as well as the adult worms, synthesize sex steroid hormones from [3H]steroid precursors and that androgens and oestrogens influence the in vitro development of the parasites. Glucocorticoids (GCs) are used to control the inflammation caused by T. solium cysticerci in the brain. These steroids stimulate oestrogen synthesis in several tissues. Since there is no information on the effect of GC on the endocrine function of cysticerci, we investigated the effect of natural and synthetic GCs on the synthesis of oestrogens in cultured T. crassiceps WFU cysticerci. The cysticerci were obtained from the peritoneal cavity of infected female BALB/c mice; the cysts were washed extensively and pre-cultured in Dulbecco's Modified Eagle's Medium (DMEM) plus antibiotics for 5 days. The parasites were further cultured with different doses of corticosterone, dexamethasone or the vehicle for 5 days. [3H]Dehydroepiandrosterone (3H-DHEA) was added to the media and the cysticerci were further incubated for 6 or 24 h. Media were then removed and the steroids ether-extracted. Aliquots of the media were seeded on silica gel plates and developed in solvent systems. Parasites incubated in the presence of 3H-DHEA synthesized [3H]androstenediol, [3H]testosterone and [3H]17ß-oestradiol ([3H]17ß-E2). The addition of 100 nm or higher corticosterone doses to the media increased [3H]17ß-E2 synthesis fourfold after 24 h. Dexamethasone also increased [3H]17ß-E2 synthesis. The experiments presented here show for the first time that corticosterone and the synthetic GC dexamethasone modulate the synthesis of oestrogens by cysticerci.

Towards the molecular characterization of the stable producer phenotype of recombinant antibody-producing NS0 myeloma cells.

The loss of heterologous protein expression is one of the major problems faced by industrial cell line developers and has been reported by several authors. Therefore, the understanding of the mechanisms involved in the generation of stable and high producer cell lines is a critical issue, especially for those processes based on long term continuous cultures. We characterized two recombinant NS0 myeloma cell lines expressing Nimotuzumab, a humanized anti-human epidermal growth factor receptor (EGFR) antibody. The hR3/H7 clone is a stable producer obtained from the unstable hR3/t16 clone. The unstable clone was characterized by a bimodal distribution of intracellular immunoglobulin staining using flow cytometry. Loss of antibody production was due to the emergence of a non-producer cell subpopulation that increased with cell generation number. Immunoglobulin heavy chain (HC) and light chain (LC) ratio (HC/LC) was lower for the unstable phenotype. Proteomic maps using two dimensional gel electrophoresis (2DE) were obtained for both clones, at initial cell culture time and after 40 generations. Fifteen proteins potentially associated with the phenomenon of production stability were identified. The hR3/H7 stable clone showed an up-regulated expression pattern for most of these proteins. The regulation of recombinant antibody production by the host NS0 myeloma cell line most likely involves simultaneously cellular processes such as DNA transcription, mRNA processing, protein synthesis and folding, vesicular transport, glycolysis and energy production, according to the proteins identified in the present proteomic study.

Phylogeny of Chaetanthera (Asteraceae: Mutisieae) reveals both ancient and recent origins of the high elevation lineages.

Penalized likelihood analysis of previously published chloroplast DNA (cpDNA) ndhF sequences suggests that the central-southern Andean genus Chaetanthera diverged ca. 16.5 million years (my) ago, well before the uplift of the Andes to their present heights. Penalized likelihood analysis based on new nuclear ribosomal DNA (rDNA) internal transcribed spacer (ITS) sequences indicates that the most relictual lineages occupy high elevation Andean habitats that did not exist until some 10my later. This result is contrary to the expectation that younger habitats should be occupied by phylogenetically younger lineages. The results are interpreted with respect to the development of aridity in lowland habitats during the Miocene and Pliocene, which presumably extinguished the lowland relatives of the high elevation taxa or, in effect, forced them upwards in search of adequate moisture. As the more northerly lineages were being displaced upward, others diversified in the mediterranean-type climate area of central Chile, giving rise to additional high elevation taxa again, at an early date, as well as lowland taxa. Some species of Chaetanthera from lowland central Chile appear as the phylogenetically youngest taxa, suggesting secondary adaptation to lowland aridity. At the same time, at least two high elevation species, Chaetanthera peruviana and Chaetanthera perpusilla, appear to have been derived recently from a lower elevation ancestor, while some middle to low elevation taxa seem to have evolved recently out of a high elevation complex. The results suggest that the younger high elevation habitats have served as both "cradle" and "museum" for Chaetanthera lineages.

[Effect of strontium on bone metabolism in hemodialysis patients]. / Efecto del estroncio sobre el metabolismo óseo en pacientes en hemodiálisis.

The objective of this study was to assess the relationship between the bone strontium content and bone histomorphometric parameters in bone biopsies from patients with chronic renal failure undergoing hemodialysis. The study was carried out in 74 illiac crest bone biopsies from patients with renal osteodystrophy from different worldwide regions (Argentina, Portugal and Spain). They were underwent to histological and histomorphometric evaluation. The bone strontium/calcium ratio was measured by quadrupole inductively coupled plasma-mass spectrometry. The samples were classified into groups according to histological criteria: hyperparathyroidism (HP), mixed (MX), osteomalacia (OM) and adynamic bone disease (ABD). Serum PTH and alkaline phosphatase before biopsy were available in most of the patients. No correlation was found between the different histomorphometric parameters and the Sr/Ca ratio. The one way ANOVA test showed statistical differences in the Sr/Ca ratio of the different histological forms (HP: 0.58 +/- 0.39; MX: 1.16 +/- 0.74; OM: 1.10 +/- 0.46; ABD: 0.91 +/- 0.40 microgram Sr/mg Ca; p < 0.003). The post-Hoc analysis showed differences between HP and MX. The biopsies having greater or equal values than 1.4 micrograms Sr/mg Ca showed higher levels of bone formation histomorphometric parameters and serum alkaline phosphatase (395 +/- 519 vs 1,022 +/- 989 UI/L, p < 0.05). Although it has been found that the biopsies with higher bone strontium had higher levels of osteoid tissue (characteristic of osteomalacia), the hypothesis of strontium-induced osteomalacia could not be demonstrated.

Efecto del estroncio sobre el metabolismo óseo en pacientes en hemodiálisis / Effect of strontium on bone histomorphometric parameters in haemodialysis patients

El objetivo de este estudio fue investigar la relación que existe entre el contenido de estroncio en el hueso y parámetros histomorfométricos óseos en biopsias de pacientes con insuficiencia renal crónica sometidos a hemodiálisis. El estudio se realizó sobre 74 biopsias óseas de cresta ilíaca de enfermos con osteodistrofia renal de distintas procedencias (Argentina, Portugal y España) que fueron sometidas a análisis histológico e histomorfométrico. Además se cuantificó estroncio y calcio en un fragmento de la biopsia mediante espectrometría de masas con fuente de plasma acoplado por inducción (ICP-MS). Las biopsias fueron clasificadas en cuatro grupos según criterios histológicos: hiperparatiroidismo (HP), enfermedad mixta (MX), osteomalacia (OM) y enfermedad ósea adinámica (EOA). También fueron medidos los niveles séricos de fosfatasa alcalina y de PTH intacta. No se encontró correlación entre los distintos parámetros histomorfométricos y la relación Sr/Ca. El análisis de varianza mostró diferencias significativas de Sr/Ca entre las distintas formas histológicas (HP: 0,58 ± 0,39; MX: 1,16 ± 0,74; OM: 1,10 ± 0,46; AD: 0,91 ± 0,40; p < 0,003), habiendo diferencias entre HP y MX (p < 0,005). Cuando se dividieron las biopsias en dos grupos de acuerdo al nivel de la relación Sr/Ca (estroncio bajo < 1,4 μg Sr/mg Ca, estroncio alto _ 1,4 μg Sr/mg Ca), se encontró que aquellas con estroncio elevado mostraron niveles más altos en parámetros de formación ósea. De los parámetros séricos analizados, solo la fosfatasa alcalina mostró valores incrementados en los pacientes con Sr elevado en el hueso (395 ± 519 vs 1.022 ± 989 UI/L). Si bien se ha encontrado que aquellas biopsias con estroncio elevado muestran niveles más altos de osteoide (compatible con osteomalacia), el grado de causalidad del mismo no ha quedado claramente establecido (AU)

The objective of this study was to assess the relationship between the bone strontium content and bone histomorphometric parameters in bone biopsies from patients with chronic renal failure undergoing hemodialysis. The study was carried out in 74 illiac crest bone biopsies from patients with renal osteodystrophy from different worldwide regions (Argentina, Portugal and Spain). They were underwent to histological and histomorphometric evaluation. The bone strontium/calcium ratio was measured by quadrupole inductively coupled plasma-mass spectrometry. The samples were classified into groups according to histological criteria: hyperparathyroidism (HP), mixed (MX), osteomalacia (OM) and adynamic bone disease (ABD). Serum PTH and alkaline phosphatase before biopsy were available in most of the patients. No correlation was found between the different histomorphometric parameters and the Sr/Ca ratio. The one way ANOVA test showed statistical differences in the Sr/Ca ratio of the different histological forms (HP: 0.58 ± 0.39; MX: 1.16 ± 0.74; OM: 1.10 ± 0.46; ABD: 0.91 ± 0.40 μg Sr/ mg Ca; p < 0.003). The post-Hoc analysis showed differences between HP and MX. The biopsies having greater or equal values than 1.4 μg Sr/mg Ca showed higher levels of bone formation histomorphometric parameters and serum alkaline phosphatase (395 ± 519 vs 1,022 ± 989 UI/L, p < 0.05) Although it has been found that the biopsies with higher bone strontium had higher levels of osteoid tissue (characteristic of osteomalacia), the hypothesis of strontium-induced osteomalacia could not be demonstrated (AU)

A phase II study of gemcitabine and cisplatin combination as induction chemotherapy for untreated locally advanced cervical carcinoma.

BACKGROUND: Cisplatin-based chemoradiation for locally advanced cervical carcinoma is now the standard of care for most patients with cervical carcinoma. However, induction chemotherapy followed by surgery, particularly with newer agents or combinations remains to be explored. This study was undertaken to evaluate the antitumor activity and toxicity of gemcitabine in combination with cisplatin for untreated locally advanced cervical carcinoma. PATIENTS AND METHODS: Open-label, single center, phase II, non-randomized study of neoadjuvant gemcitabine plus cisplatin. Forty-one patients with histologic diagnosis of cervical carcinoma, with no previous treatment and staged as IB2 to IIIB, were treated with three 21-day courses of cisplatin 100 mg/m2 day I and gemcitabine 1000 mg/m2 days 1 and 8, followed by locoregional treatment with either surgery or concomitant chemoradiation. Response and toxicity were evaluated before each course and at the end of chemotherapy. RESULTS: All patients were evaluated for toxicity and 40 for response. The overall objective response rate was 95% (95% confidence interval (CI): 88%-100%) being complete in 3 patients (7.5%) and partial in 35 (87.5%). A complete pathological response was found in 6 (26%) of the 23 patients that underwent surgery. Granulocytopenia grades 3-4 occurred in 13.8% and 3.4% of the courses, respectively, whereas non-hematological toxicity was mild. CONCLUSIONS: Induction chemotherapy with the combination of gemcitabine and cisplatin is highly active for untreated cervical cancer patients and has an acceptable toxicity profile.

Type A behavior pattern, sex, and social support.

Low social support has been suggested as a contributing factor to cardiac disease in some individuals with Type A Behavior Pattern. A number of studies have focused on the relationship between social support and Type A traits in subjects without cardiac disease as a means of evaluating this hypothesis; however, results of these studies have been confusing. In the current study, involving samples of adolescents and working adults, there was a significant positive correlation between Type A scores and satisfaction with social support in men but not in women.

Effects of infantile thymectomy on ovarian functions and gonadotrophin-induced ovulation in prepubertal mice: role of thymulin.

The effects of thymectomy performed on 10-day-old (Tx-10) mice on spontaneous puberty and the ovulatory response induced by gonadotrophin treatment were analysed, together with the effects of thymulin replacement from 10 days of age. Infantile thymectomy induced a delay of puberty, a decrease in serum 17beta-oestradiol concentration and a reduced total number of follicles. Injection of thymulin (12 ng/g body weight) to Tx-10 mice resulted in an earlier onset of puberty, a decrease in the weights of ovaries and uterus, and an increase in serum 17beta-oestradiol concentrations. In control and Tx-10 mice, treatment with pregnant mare serum gonadotrophin (PMSG) (5 IU) at 25 days of age resulted in ovulation and the numbers of ova shed by ovulating animals were similar. When the animals were injected with 1 IU PMSG ovulation did not occur. In Tx-10 mice, both 1 and 5 IU PMSG increased the number of follicles to values similar to those observed in the controls. In Tx-10 mice the sequential injection of PMSG (1 IU) and human chorionic gonadotrophin (hCG) (3 IU) resulted in ovulation, but the number of ova shed was lower than in controls. When these animals were injected daily with thymulin, an increase in the number of ova shed and serum 17beta-oestradiol concentrations was observed. The uterine weight of Tx-10 mice was always significantly reduced in response to gonadotrophin treatment. Thymulin injection in PMSG-hCG-treated Tx-10 mice provoked a significant increase in uterine weight. The results suggest that the presence of the thymus after the neonatal period is necessary to normal ovarian development and function. The increase in gonadotrophin-induced ovarian response produced by thymulin replacement indicates that this peptide has a role in this process as one of the connecting signals between thymus and ovaries.

A Theoretical-Experimental Study on the Structure and Activity of Certain Quinolones and the Interaction of Their Cu(II)-Complexes on a DNA Model.

Theoretical electronic Structure methods have been employed to study the structure and activity of certain (free) quinolones and the interaction of their Cu(II)-complexes on a DNA model (Rhodamine 6G (rhod)). As a manner of assessing the generated geometries, the nalidixic acid geometrical parameters obtained were tested against the crystallographic ones and it was found that the average error in the calculated geometries is small. The present study allows us to (1) Rationalize the observed differences in antibiotic activities through their electronic hardnesses. (2) Suggest a plausible mechanism of action for these drugs through formation of a reactive intermediate (or carrier) which would consist of a quinolone anion coordinated to an adequate metal center (Cu(II) in this study). (3) We find that, through this model of DNA (modeled with rhod) the interaction seems to be mediated by an effective pi-pi stacking. (4) Finally, an in vitro experiment was designed so that the intercalation process in DNA could be experimentally modeled as well. The quenching of the rhod fluorescence is proportional to the strength of the Cu(II)-complex-rhod interaction and therefore provides a quantitative measurement of the "intercalating" capacity of the quinolones and their copper complexes. These results agree well with the theoretical total adduct formation energies.

Effects of thymulin on spontaneous puberty and gonadotrophin-induced ovulation in prepubertal normal and hypothymic mice.

The effects of thymulin administration beginning on days 19 or 24 of age on spontaneous puberty and gonadotrophin-induced ovulation were analysed in female normal and hypothymic mice. In normal and hypothymic mice, the daily administration of thymulin at 24 days of age resulted in a delay in the age of vaginal opening, with an increase in serum progesterone levels. Normal mice treated with 200 ng thymulin beginning on day 19 of age and injected with pregnant mare serum gonadotrophin (PMSG) 24 h later had an increase in ovulation rate, number of ova shed and weight of the ovaries. None of the hypothymic mice treated with thymulin on day 19 and PMSG on day 20 ovulated. PMSG treatment on day 25 induced ovulation in hypothymic mice. When these animals were injected previously with 200 ng thymulin, the number of ova shed by ovulating animals was lower than in PMSG-treated animals. Administration of thymulin and sequential injection of PMSG and human chorionic gonadotrophin 54 h later resulted in an increase in ovulatory response in comparison with those receiving only PMSG. The results suggest that thymulin plays a role in the regulation of spontaneous puberty through its effects on adrenal and ovarian endocrine functions. The increase in the ovarian PMSG response-treated animals, previously given thymulin, showed that this thymic hormone participates in the regulation of gonadotrophin secretion mechanisms and seems to be dose- and age-dependent. In hypothymic mice, neuroendocrine mechanisms regulating puberty are different from those of normal mice.

Estimation of temperature and precipitation from morphological characters of dicotyledonous leaves.

The utility of regression and correspondence models for deducing climate from leaf physiognomy was evaluated by the comparative application of different predictive models to the same three leaf assemblages. Mean annual temperature (MAT), mean annual precipitation (MAP), and growing season precipitation (GSP) were estimated from the morphological characteristics of samples of living leaves from two extant forests and an assemblage of fossil leaves. The extant forests are located near Gainesville, Florida, and in the Florida Keys; the fossils were collected from the Eocene Clarno Nut Beds, Oregon. Simple linear regression (SLR), multiple linear regression (MLR), and canonical correspondence analysis (CCA) were used to estimate temperature and precipitation. The SLR models used only the percentage of species having entire leaf margins as a predictor for MAT and leaf size as a predictor for MAP. The MLR models used from two to six leaf characters as predictors, and the CCA used 31 characters. In comparisons between actual and predicted values for the extant forests, errors in prediction of MAT were 0.6°-5.7°C, and errors in prediction of precipitation were 6-89 cm (=6-66%). At the Gainesville site, seven models underestimated MAT and only one overestimated it, whereas at the Keys site, all eight models overestimated MAT. Precipitation was overestimated by all four models at Gainesville, and by three of them at the Keys. The MAT estimates from the Clarno leaf assemblage ranged from 14.3° to 18.8°C, and the precipitation estimates from 227 to 363 cm for MAP and from 195 to 295 cm for GSP.

Metabolism and toxicity of 2-bromo-(diglutathion-S-yl)-hydroquinone and 2-bromo-3-(glutathion-S-yl)hydroquinone in the in situ perfused rat kidney.

2-Br-(diglutathion-S-yl)hydroquinone (2-Br-(diGSyl)HQ) is a potent nephrotoxicant, causing glucosuria, enzymuria, proteinuria, elevations in blood urea nitrogen, and severe histological alterations to renal proximal tubules at doses of 10-15 mumol/kg. In contrast, 2-Br-3-(glutathion-S-yl)hydroquinone (2-Br-3-(GSyl)HQ) is substantially less nephrotoxic than 2-Br-(diGSyl)HQ and requires a dose of at least 50 mumol/kg to cause modest elevations in blood urea nitrogen concentrations. The reason or reasons for this difference in potency is unclear, but since inhibition of renal gamma-glutamyl transpeptidase (gamma-GT) prevents 2-Br-(diGSyl)HQ-mediated nephrotoxicity, metabolism of these conjugates by the kidney must play an important role. To address this question we have compared the metabolism and toxicity of 2-Br-(diGSyl)HQ and 2-Br-3-(GSyl)HQ in the in situ perfused rat kidney (ISPRK). Following infusion of 20 mumol 2-Br-3-(GSyl)HQ into the right renal artery of male Sprague Dawley rats, a total of 23.5 +/- 1.9% (mean +/- SE) of the dose was accounted for in urine and bile over a period of 180 min. 2-Bromo-3-(cystein-S-yl)hydroquinone and 2-bromo-3-(N-acetylcystein-S-yl)hydroquinone were identified in urine, and unchanged 2-Br-3-(GSyl)HQ was identified in urine and bile. The product arising from the oxidative cyclization of 2-bromo-3-(cystein-S-glycine)hydroquinone, 2H-(3-glycine)-7-hydroxy-8-bromo-1,4-benzothiazine, was also identified in urine.(ABSTRACT TRUNCATED AT 250 WORDS)

Formation and mineralization of extracellular matrix secreted by an immortal human osteoblastic cell line: modulation by tumor necrosis factor-alpha.

Tumor necrosis factor-alpha (TNF-alpha), a 17-kDa cytokine produced by stimulated macrophages/monocytes, modulates the functions of a variety of cells and has been shown to induce bone resorption in vitro. However, the effects that TNF-alpha may have on the process of bone formation are not completely understood. In order to study the effects of TNF-alpha on matrix development and mineralization, we utilized a human osteoblastic cell line, HOS TE85. Our results show that HOS TE85, which has been shown to be responsive to hormones active on normal osteoblasts, forms an extensive extracellular matrix (ECM) that mineralizes during extended culture. Treatment during the development of the matrix with TNF-alpha has little effect on cell number and DNA synthesis, showing thereby that TNF-alpha is not cytotoxic to the cells. However, TNF-alpha inhibits the formation of alkaline phosphatase (AP)-positive foci in a dose-dependent manner at concentrations of 0.1-10 ng/ml. TNF-alpha treatment caused a significant decrease in the incorporation of collagen into the developing matrix. In addition, TNF-alpha treatment resulted in a significant decrease in the synthesis of AP by HOS TE85 cells during the process of ECM formation and resulted in a pronounced lack of mineralization of the ECM. These results indicate that TNF-alpha may be acting as an uncoupler by decreasing the synthesis and incorporation of proteins required for bone formation, and inhibiting matrix formation and mineralization in vitro.

[Superficial gastric cancer in Barcelona. Importance of the detection of severe epithelial dysplasia in biopsies obtained under endoscopic control in the diagnosis of early gastric cancer]. / Cáncer gástrico superficial en Barcelona. Importancia de la detección de displasia epitelial acusada en las biopsias obtenidas bajo control endoscópico para el diagnóstico del cáncer gástrico incipiente.

BACKGROUND: Severe mucosal dysplasia (SMD) in endoscopic gastric biopsies is a controversial lesion because some authors consider it as an histologic lesion associated to superficial gastric cancer (SCG). This study is aimed to asses the prevalence of SCG in Barcelona; if the presence of SMD is associated to SCG and, the clinical and endoscopic manifestations which induced the diagnosis. METHODS: We studied a total of 4,800 patients who had been submitted to gastroscopy during 1.5 years. A total of 79 patients suffered gastric cancer, 56 of them were submitted to resection. Ten of these 56 patients (17%) had SCG. The most frequent first clinical manifestation of SCG was gastrointestinal haemorrhage. RESULTS: Endoscopic aspect suggested malignancy in five cases. Biopsies during endoscopy allow diagnosis of gastric cancer in five. In four histologic studies showed SMD. In the tenth patient, the first endoscopy with biopsies showed a gastric peptic ulcer. A later endoscopic control showed the persistence of the macroscopically benign lesion but one among seven biopsies showed adenocarcinoma. The four patients with SMD had gastric cancer in further endoscopic procedures, except in one which was operated after two endoscopies demonstrating SMD. CONCLUSIONS: This study suggests: 1) prevalence of SGC in Barcelona is similar than the other European countries. 2) haemorrhage is a frequent first manifestation of SCG, and 3) the finding of SMD in endoscopic biopsy strongly suggests the presence of SCG in the stomach.