Microsomal 11a-hydroxylation of progesterone in Image : Part I: Characterization of the hydroxylase system

Microsomes (105,000xg sediment) prepared from induced cells of Image was found to hydroxylate progesterone to 11a-hydroxyprogesterone (11a-OHP) in high yields (85-90% in 30 min.) in the presence of NADPH and O2. The pH optimum for the hydroxylase was found to be 7.7. However, for the isolation of active microsomes grinding of the mycelium should be carried out at pH 8.3. Metyrapone, carbon monoxide, SKF-525A, p-CMB and N-methyl maleimide inhibited the hydroxylase activity indicating the involvement of cytochrome P-450 system. The inhibition of the hydroxylase by cytochrome Image and the presence of high levels of NADPH-cytochrome Image reductase in induced microsomes suggest that the reductase could be one of the components in the hydroxylase system.

Solution NMR studies of acetohydroxy acid synthase I: Identification of the sites of inter-subunit interactions using multidimensional NMR methods

The novel multidomain organization in the multimeric Escherichia coli AHAS I (ilvBN) enzyme has been dissected to generate polypeptide fragments. These fragments when cloned, expressed and purified reassemble in the presence of cofactors to yield a catalytically competent enzyme. Structural characterization of AHAS has been impeded due to the fact that the holoenzyme is prone to dissociation leading to heterogeneity in samples. Our approach has enabled the structural characterization using high-resolution nuclear magnetic resonance methods. Near complete sequence specific NMR assignments for backbone H-N, N-15, C-13 alpha and C-13(beta) atoms of the FAD binding domain of ilvB have been obtained on samples isotopically enriched in H-2, C-13 and N-15. The secondary structure determined on the basis of observed C-13(alpha) secondary chemical shifts and sequential NOEs indicates that the secondary structure of the FAD binding domain of E. coli AHAS large Subunit (ilvB) is similar to the structure of this domain in the catalytic subunit of yeast AHAS. Protein-protein interactions involving the regulatory subunit (ilvN) and the domains of the catalytic subunit (ilvB) were studied using circular dichroic and isotope edited solution nuclear magnetic resonance spectroscopic methods. Observed changes in circular dichroic spectra indicate that the regulatory subunit (ilvN) interacts with ilvB alpha and ilvB beta domains of the catalytic subunit and not with the ilvB gamma domain. NMR chemical shift mapping methods show that ilvN binds close to the FAD binding site in ilvB beta and proximal to the intrasubunit ilvB alpha/ilvB beta domain interface. The implication of this interaction on the role of the regulatory subunit oil the activity of the holoenzyme is discussed. NMR studies of the regulatory domains show that these domains are structured in solution. Preliminary evidence for the interaction of ilvN with the metabolic end product of the pathway, viz., valine is also presented.

Rights of Portugal, in reference to Great Britain and the question of the slave trade, or, the manifesto and protest of the weak, against the ingratitude, oppression, and violence of the strong ...

Enquanto os políticos e diplomatas polemizavam e agiam, Thomas Buxton publicava, depois de viajar ao Brasil e ao Caribe, o mais completo levantamento sobre o tráfico negreiro jamais feito até então.

A quantitative analysis of the phytoplankton of the Gulf of Panama: I. Results of the regional phytoplankton surveys during July and November 1957 and March 1958

ENGLISH: 1. Quantitative phytoplankton samples were collected by the Inter-American Tropical Tuna Commission at the surface and ten meters in the Gulf of Panama, as follows: a) 18-21 March, 1958 (31 stations)-during the height of the upwelling season, b) 10-12 July, 1957 (10 stations)-during the transition to the rainy season at a time when mild upwelling winds reappear, c) 7-8 November, 1957 (15 stations)-during the height of the rainy season. 2. Maximum phytoplankton populations occurred during the upwelling season, followed by a considerable decline during July, and a further Subsidence during November. 3. A remarkable regional uniformity in species composition was observed during the surveys despite regional differences in growth conditions. Diatoms overwhelmingly dominated the communities. 4. During all surveys, the innermost regions, generally north of 8°30'N, were the most productive. The least productive areas were in the offing of San Miguel Bay and Parita Bay, suggesting that nutrient accretion via runoff is inadequate to sustain sizeable autotrophic plant populations in those regions. 5. During all surveys, phytoplankton growth appeared to be limited by nutrient availability. 6. During all surveys, phytoplankton growth appeared to be related to depth of the water column. 7. Although below average rainfall contributed to unusually favorable growth conditions (reduced stability, increased transparency and, presumably, nutrient reserves) during the November survey relative to November 1955 and 1956 at 8°45'N, 79°23'W, the anticipated heightened phytoplankton response was not observed. 8. During the November survey, the local diatom responses and their regional fluctuations could be satisfactorily related to the accompanying surface salinity conditions. However, this correspondence is undoubtedly attributable to factors associated with the observed salinity levels, probably nutrients, rather than salinity directly. 9. Unusually warm conditions occurred during the March survey, attributable to considerably weaker upwelling winds than normally occurring then, which contributed to a considerably lower standing crop and a retardation in succession of three to five weeks relative to that observed during 1955-1957 at 8°45'N, 79°23'W in the Gulf of Panama. 10. During the March survey, a well defined inverse relationship existed between mean temperature and mean diatom abundance in the upper ten meters, and between transparency and mean diatom abundance. A direct relationship occurred between surface salinity and mean diatom abundance in the upper ten meters. These relationships are interpreted to indicate that diatom abundance primarily reflected the nutrient concentrations associated with a given upwelling intensity, rather than describing casual relationships. 11. The survey results indicate that the phytoplankton dynamics observed at 8°45'N, 79°23'W from November, 1954 through May, 1957 are generally representative of the Gulf of Panama. 12. The following new forms, to be described in a later publication, were observed during the surveys: Actinoptychus undulatus f. catenata n.f., Asterionella japonica f. tropicum n.f., Leptocylindrus maximus n. sp., Skeletonema costatum f. tropicum n.f. SPANISH: 1. La Comisión Interamericana del Atun Tropical recolectó en el Golfo de Panama muestras cuantitativas de fitoplancton en la superficie y a los diez metros, como sigue: a) Del 18 al 21 de marzo de 1958 (31 estaciones)-durante el maximum de la estación de afloramiento. b) Del 10 al 12 de julio de 1957 (10 estaciones)-durante la epóca de transición a la estación lluviosa cuando reaparecen los vientos ligeros que causan el afloramiento. c) Del 7 al 8 de noviembre de 1957 (15 estaciones)-durante el maximum de la estación lluviosa. 2. Las poblaciones maximas de fitoplancton aparecieron durante la estación de afloramiento, seguido por una considerable disminución durante el mes de julio y una calma durante noviembre. 3. Durante la investigación se observó una remarcable uniformidad regional en la composición de las especies a pesar de las diferencias regionales en las condiciones de crecimiento. Las diatomeas predominaban en gran numero en las comunidades. 4. Durante todas las investigaciones, las regiones mas cerca de la costa, generalmente al norte de los 8°30'N, eran las mas productivas. Las areas menos productivas fueron las mar afuera de las Bahias de San Miguel y Parita, lo que sugiere que el aumento en las sales nutritivas causado por las escorrentias es inadecuado para sostener poblaciones grandes de plantas autotróficas en estas regiones. 5. Durante todas las investigaciones, el crecimiento del fitoplancton parecio estar limitado por la disponibilidad de las. sales nutritivas. 6. Durante todas las investigaciones el crecimiento del fitoplancton parecio estar relacionado con la profundidad de la columna de agua. 7. Aunque las precipitacion por debajo del promedio normal contribuyo a condiciones desusadamente favorables de crecimiento (estabilidad reducida, aumento de la transparencia y, presumiblemente, de la reserva de sales nutritivas) durante la investigación de noviembre en relación a noviembre de 1955 y de 1956 en los 8°45'N, 79°23'W, no se observo-la alta reacción de fitoplancton que se esperaba. 8. Durante la investigación de noviembre, las reacciones locales de las diatomeas y sus fluctuaciones regionales pudieron relacionarse en forma satisfactoria con condiciones asociadas con la salinidad de la superficie. Sin embargo, esta correspondencia puede atribuirse sin duda a factores asociados con los niveles observados de salinidad, probablemente con las sales nutritivas, en lugar de directamente con la salinidad. 9. Condiciones calurosas no comunes ocurrieron durante la investigación de marzo, las que pueden atribuirse a que los vientos que ocasionan el afloramiento fueran mas debiles que los normales, lo que contribuyó a que la cosecha estable fuera considerablemente mas baja y a la demora de tres a cinco semanas en la sucecion relativa a la que se observó durante 1955-1957 en los 8°45'N, 8°23'W, en el Golfo de Panama. 10. Durante la investigación de marzo, existió una relación inversa bien definida entre la temperatura y la abundancia media de las diatomeas en los diez metros superiores, y entre la transparencia y la abundancia media de las diatomeas. Una relación directa ocurrio entre la salinidad de superficie y la abundancia media de las diatomeas en los diez metros superiores. Estas relaciones se interpretan como indicadoras de que la abundancia de diatomeas refleja primeramente las concentraciones de las sales nutritivas asociadas con una intensidad de afloramiento dada, en lugar de describir relaciones causales. 11. Los resultados de la investigacion indican que la dinamica del fitoplancton observada en los 8°45'N, 79°23'W, desde noviembre de 1954 a mayo de 1957, es generalmente representativa del Golfo de Panama. 12. Durante las investigaciones se observaron las siguientes formas nuevas, las que seran descritas en una publicación posterior: Actinoptychus undulatus f. catenata n.f., Asterionella japonica f. tropicum n.f., Leptocylindrus maximus n. sp., Skeletonema costatum f. tropicum n.f.

Isothermal crystallization kinetics of PEO in poly(ethylene terephthalate)-poly(ethylene oxide) segmented copolymers. I. Effect of the sofi-block length

The isothermal crystallization kinetics of poly(ethylene oxide) (PEO) block in two poly(ethylene terephthalate) (PET)-PEO segmented copolymers was studied with differential scanning calorimetry. The Avrami equation failed to describe the overall crystallization process, but a modified Avrami equation, the Q equation, did. The crystallizability of the PET block and the different lengths of the PEO block exerted strong influences on the crystallization process, the crystallinity, and time final morphology of the PEO block. The mechanism of nucleation and the growth dimension of the PEG block were different because of the crystallizability of time PET block and the compositional heterogeneity. The crystallization of the PEO block was physically constrained by the microstructure of time PET crystalline phase, which resulted in a lower crystallization rate. However, this influence became weak with the increase in the soft-block length. (C) 2000 John Wiley & Sons, Inc.

Isobutane/butene alkylation over supported heteropoly acid catalysts: I. Influence of the structure of silica

Catalysts consisting of heteropoly acids (HPAs) supported on different silica and mesoporous molecular sieves have been prepared by impregnation and the sol-gel method, respectively, and their catalytic behavior in fixed-bed alkylation of isobutane with butene has been investigated. The activity, selectivity and stability of the supported-HPA catalysts could be correlated with the surface acidity of the catalysts, the structure of supports as well as the time on stream (TOS). In the fixed-bed reactor, the acidity of the heteropoly acid is favorable to the formation of dimerization products (C-8(=)); especially, the pore size of supports was seen to have an important effect on activity and product distribution of the catalysts. Contrary to the traditional solid-acid catalysts, the supported-HPA catalysts own an excellent stability for alkylation, which makes it possible for these supported catalysts to replace the liquid-acid catalysts used in industry.

A randomised, phase II trial of the DNA-hypomethylating agent 5-aza-2'-deoxycytidine (decitabine) in combination with carboplatin vs carboplatin alone in patients with recurrent, partially platinum-sensitive ovarian cancer.

Background: Our previous laboratory and clinical data suggested that one mechanism underlying the development of platinum resistance in ovarian cancer is the acquisition of DNA methylation. We therefore tested the hypothesis that the DNA hypomethylating agent 5-aza-2'-deoxycytodine (decitabine) can reverse resistance to carboplatin in women with relapsed ovarian cancer.

Methods: Patients progressing 6-12 months after previous platinum therapy were randomised to decitabine on day 1 and carboplatin (AUC 6) on day 8, every 28 days or carboplatin alone. The primary objective was response rate in patients with methylated hMLH1 tumour DNA in plasma.

Results: After a pre-defined interim analysis, the study closed due to lack of efficacy and poor treatment deliverability in 15 patients treated with the combination. Responses by GCIG criteria were 9 out of 14 vs 3 out of 15 and by RECIST were 6 out of 13 vs 1 out of 12 for carboplatin and carboplatin/decitabine, respectively. Grade 3/4 neutropenia was more common with the combination (60% vs 15.4%) as was G2/3 carboplatin hypersensitivity (47% vs 21%).

Conclusions: With this schedule, the addition of decitabine appears to reduce rather than increase the efficacy of carboplatin in partially platinum-sensitive ovarian cancer and is difficult to deliver. Patient-selection strategies, different schedules and other demethylating agents should be considered in future combination studies.

A Pension for Injured Victims of the Troubles: Reparations or Reifying Victim Hierarchy?

Reparations have been often used victim-centred measures to redress both private harm and gross violations of human rights. However, with the increasing occurrence of internal armed conflict and political violence, identities of victims and perpetrators in protracted conflicts can become blurred for some individuals. In countries like Peru and Northern Ireland that have suffered protracted violence, victimhood has been contested around which individuals are seen as innocent and deserving to exclude any members of non-state armed groups from claiming reparations. This article explores the issue of a proposed bill on a pension for injured victims of the Troubles in Northern Ireland. It identifies that there is no consistent state practice or human rights jurisprudence in this area, but instead offers a more complex approach through four models that can grapple with the seeming diametrically opposed victimhood and responsibility, by including victimised-perpetrators in reparations programmes such as that proposed for a pension of seriously injured victims in Northern Ireland.

A Communication Intervention for Training Southern European Oncologists to Recognize Psychosocial Morbidity in Cancer. I - Development of the Model and Preliminary Results on Physicians' Satisfaction

BACKGROUND: The detection of psychosocial distress is a significant communication problem in Southern Europe and other countries. Work in this area is hampered by a lack of data. Because not much is known about training aimed at improving the recognition of psychosocial disorders in cancer patients, we developed a basic course model for medical oncology professionals. METHODS: A specific educational and experiential model (12 hours divided into 2 modules) involving formal teaching (ie, journal articles, large-group presentations), practice in small groups (ie, small-group exercises and role playing), and discussion in large groups was developed with the aim of improving the ability of oncologists to detect emotional disturbances in cancer patients (ie, depression, anxiety, and adjustment disorders). RESULTS: A total of 30 oncologists from 3 Southern European countries (Italy, Portugal, and Spain) participated in the workshop. The training course was well accepted by most participants who expressed general satisfaction and a positive subjective perception of the utility of the course for clinical practice. Of the total participants, 28 physicians (93.3%) thought that had they been exposed to this material sooner, they would have incorporated the techniques received in the workshop into their practices; 2 participants stated they would likely have done so. Half of the doctors (n = 15) believed that their clinical communication techniques were improved by participating in the workshop, and the remaining half thought that their abilities to communicate with cancer patients had improved. CONCLUSIONS: This model is a feasible approach for oncologists and is easily applicable to various oncology settings. Further studies will demonstrate the effectiveness of this method for improving oncologists skills in recognizing emotional disorders in their patients with cancer.