Trans-spliced Heat Shock Protein 90 Modulates Encystation in Giardia lamblia

Background: Hsp90 from Giardia lamblia is expressed by splicing of two independently transcribed RNA molecules, coded by genes named HspN and HspC located 777 kb apart. The reasons underlying such unique trans-splicing based generation of GlHsp90 remain unclear. Principle Finding: In this study using mass-spectrometry we identify the sequence of the unique, junctional peptide contributed by the 5' UTR of HspC ORF. This peptide is critical for the catalytic function of Hsp90 as it harbours an essential ``Arg'' in its sequence. We also show that full length GlHsp90 possesses all the functional hall marks of a canonical Hsp90 including its ability to bind and hydrolyze ATP. Using qRT-PCR as well as western blotting approach we find the reconstructed Hsp90 to be induced in response to heat shock. On the contrary we find GlHsp90 to be down regulated during transition from proliferative trophozoites to environmentally resistant cysts. This down regulation of GlHsp90 appears to be mechanistically linked to the encystation process as we find pharmacological inhibition of GlHsp90 function to specifically induce encystation. Significance: Our results implicate the trans-spliced GlHsp90 from Giardia lamblia to regulate an essential stage transition in the life cycle of this important human parasite.

Heat shock protein 90 as a potential drug target against surra

Trypanosomiasis is caused by Trypanosoma species which affect both human and animal populations and pose a major threat to developing countries. The incidence of animal trypanosomiasis is on the rise. Surra is a type of animal trypanosomiasis, caused by Trypanosoma evansi, and has been included in priority list B of significant diseases by the World Organization of Animal Health (OIE). Control of surra has been a challenge due to the lack of effective drugs and vaccines and emergence of resistance towards existing drugs. Our laboratory has previously implicated Heat shock protein 90 (Hsp90) from protozoan parasites as a potential drug target and successfully demonstrated efficacy of an Hsp90 inhibitor in cell culture as well as a pre-clinical mouse model of trypanosomiasis. This article explores the role of Hsp90 in the Trypanosoma life cycle and its potential as a drug target. It appears plausible that the repertoire of Hsp90 inhibitors available in academia and industry may have value for treatment of surra and other animal trypanosomiasis.

Functional characterization of heat-shock protein 90 from Oryza sativa and crystal structure of its N-terminal domain

Heat-shock protein 90 (Hsp90) is an ATP-dependent molecular chaperone that is essential for the normal functioning of eukaryotic cells. It plays crucial roles in cell signalling, cell-cycle control and in maintaining proteome integrity and protein homeostasis. In plants, Hsp90s are required for normal plant growth and development. Hsp90s are observed to be upregulated in response to various abiotic and biotic stresses and are also involved in immune responses in plants. Although there are several studies elucidating the physiological role of Hsp90s in plants, their molecular mechanism of action is still unclear. In this study, biochemical characterization of an Hsp90 protein from rice (Oryza sativa; OsHsp90) has been performed and the crystal structure of its N-terminal domain (OsHsp90-NTD) was determined. The binding of OsHsp90 to its substrate ATP and the inhibitor 17-AAG was studied by fluorescence spectroscopy. The protein also exhibited a weak ATPase activity. The crystal structure of OsHsp90-NTD was solved in complex with the nonhydrolyzable ATP analogue AMPPCP at 3.1 angstrom resolution. The domain was crystallized by cross-seeding with crystals of the N-terminal domain of Hsp90 from Dictyostelium discoideum, which shares 70% sequence identity with OsHsp90-NTD. This is the second reported structure of a domain of Hsp90 from a plant source.

Heat Shock Protein 90 regulates encystation in Entamoeba

Enteric protozoan Entamoeba histolytica is a major cause of debilitating diarrheal infection worldwide with high morbidity and mortality. Even though the clinical burden of this parasite is very high, this infection is categorized as a neglected disease. Parasite is transmitted through feco-oral route and exhibit two distinct stages namely - trophozoites and cysts. Mechanism and regulation of encystation is not clearly understood. Previous studies have established the role of Heat shock protein 90 (Hsp90) in regulating stage transition in various protozoan parasites like Giardia, Plasmodium, Leishmania, and Toxoplasma. Our study for the first time reports that Hsp90 plays a crucial role in life cycle of Entamoeba as well. We identify Hsp90 to be a negative regulator of encystation in Entamoeba. We also show that Hsp90 inhibition interferes with the process of phagocytosis in Entamoeba. Overall, we show that Hsp90 plays an important role in virulence and transmission of Entamoeba.

Effect of substrates and surfactants over the evolution of crystallographic texture of nanostructured ZnO thin films deposited through microwave irradiation

In spite of intense research on ZnO over the past decade, the detailed investigation about the crystallographic texture of as obtained ZnO thin films/coatings, and its deviation with growth surface is scarce. We report a systematic study about the orientation distribution of nanostructured ZnO thin films fabricated by microwave irradiation with the variation of substrates and surfactants. The nanostructured films comprising of ZnO nanorods are grown on semiconductor substrates such as Si(100), Ge(100)], conducting substrates (ITO-coated glass, Cr coated Si), and polymer coated Si (PMMA/Si) to examine the respective development of crystallographic texture. The ZnO deposited on semiconductor substrates yieldsmixed texture, whereas c-axis oriented ZnO nanostructured films are obtained by conducting substrate, and PMMA coated Si substrates. Among all the surfactants, nanostructured film produced by using the lower molecular weight of polymeric surfactants (polyvinylpyrrolidone) shows a stronger (0002) texture, and that can be tuned to (10 - 10) by increasing the molecular weight of the surfactant. The strongest basal pole is achieved for the ZnO deposited on PMMA coated Si as substrate, and cetyl-trimethyl ammonium bromide as cationic surfactant. The texture analysis is carried out by X-ray pole figure analysis using the Schultz reflection method. (C) 2015 Elsevier B.V. All rights reserved.

First Structural View of a Peptide Interacting with the Nucleotide Binding Domain of Heat Shock Protein 90

The involvement of Hsp90 in progression of diseases like cancer, neurological disorders and several pathogen related conditions is well established. Hsp90, therefore, has emerged as an attractive drug target for many of these diseases. Several small molecule inhibitors of Hsp90, such as geldanamycin derivatives, that display antitumor activity, have been developed and are under clinical trials. However, none of these tested inhibitors or drugs are peptide-based compounds. Here we report the first crystal structure of a peptide bound at the ATP binding site of the N-terminal domain of Hsp90. The peptide makes several specific interactions with the binding site residues, which are comparable to those made by the nucleotide and geldanamycin. A modified peptide was designed based on these interactions. Inhibition of ATPase activity of Hsp90 was observed in the presence of the modified peptide. This study provides an alternative approach and a lead peptide molecule for the rational design of effective inhibitors of Hsp90 function.

Comportamiento agroindustrial de veinticinco variedades de caña de azúcar (Saccharum sp. hibrido) en comparación con la variedad L 68-90 en caña planta

En el ciclo 90-91 se llevó a cabo el presente trabajo en el Ingenio Javier Guerra Báez con el objetivo de determinar el comportamiento agroindustrial de 25 cultivares de caña de azúcar en comparación con la variedad L 68-90. El experimento se sembró el 21 de febrero de 1990 en un suelo franco de la serie Nandaime, la cosecha se realizó a los 12 meses de edad el 21 de febrero de 1991. El diseño utilizado fue látice triple (5 x 5) y las variables estudiadas fueron: Germinación. Población, altura de tallos, diámetro de tallos, peso promedio de los tallos, rendimiento agrícola. rendimiento industrial y rendimiento agroindustrial Los datos fueron sometidos a un análisis de varianza y prueba de rangos múltiples de Duncan al 5% de significancia. Los mejores comportamientos para cada variable en estudio los obtuvieron los siguientes cultivares: Germinación: RB 73-2727, Cp 70-321, RB 77-3720, RB 73-2223, RB 73-9735 y RB 76-5288; Población: Sp 70-1423, Cp 70-321 RB 73-1012. Cp 71-6180. RB 73-1714. RB 76-5288. RB 73-2727, Cp 72-1210 y C 87-51; Longitud de tallo: RB 73-429. Cp 70-1527. RB 73-9735. RB 73-1012. Q 96 MEX 69-420 y MEX 68-P23; DiáMetro de los tallos: RB 73-2223, Cp 74-383. MEX 53-473, RB 76-5288. RB 73-9735. RB 73-5220. RB 73-9953. Cp70-1527, MEX 56-476 y MEX 68-p23; Peso promedio de los tallos: MEX 68- p23. RB 73-9735. RB 73-2223. CP 70-1527, RB 73-429. MEX 53-473. Q 96. MEX 69-420 y MEX 56-476;Rendimiento agrícola: Cp 71 -6180. MEX 68- P23. X 69-420. RB 73-1012. Sp 70-1423. RB 76-5288 y RB 73-9735; Rendimiento industrial Q 96, Cp 70-321, Cp 72-1210, MEX 68 P23 y Cp71-6180; Rendimiento agroindustrial Cp 71-6180.MEX 68-P23. Cp 70-321. RB 76-5288, Cp 72-1210. MEX69-420, RB 73-1012 Y Q96. Las variedades que presentaron enfermedades fueron: C 87-51, Cp 71-6180, Cp 70-1527, RB 73-2223. RB 73-2727. L '8-90, Cp 74-383, Sp 70-1284, RB 73-1714 y RB 73-429.

Evaluación agro-industrial de veinticinco variedades de caña de azúcar (Saccharum sp. hibrido), en comparación con la variedad L 68-90, en planta y retoño 1

El presente estudio corresponde a la evaluación de dos años de producción (90-91;91-92), de 25 variedades de caña de azúcar en comparación con la variedad L 68-90, establecido en el ingenio " Julio Buitrago Urroz ", para determinar su comportamiento agro­ industrial y seleccionar las mejores variedades para una segunda fase de selección. El ensayo se sembró 24 de marzo de 1990, la cosecha de planta se realizó el 21 de marzo de 1991, y el retoño 1 se cosecho el 21 de febrero de 1992. El diseño utilizado fue el de látice triple (5 x 5) parcialmente equilibrado y los parámetros estudiados fueron: Germinación, Población, Altura, Diámetro de los tallos, Peso promedio de los tallos, Rendimiento agrícola, Rendimiento industrial, y Rendimiento agro-industrial. Cada una de las variables fueron evaluadas mediante un análisis de varianza y a la prueba de rango múltiple de Duncan a un 5% de significación. Los resultados obtenidos fueron los siguientes: Germinación; Se obtuvieron diferencias altamente significativas, las mejores variedades son: Cp 70-321, Cp 70-1527, RB 73-9735, Mex 68-P23, RB 73-1012, Mex 53-473. Población; Se presentaron diferencias altamente significativas, resultando los mejores genotipos: Rb 73-2727, Cp 70-321, Sp 70-1423. Altura; Se determinó una alta diferencia significativa entre los cultivares destacándose: Sp 72-4790, RB 76-5288, RB 73-429. Diámetro de tallo; Se obtuvo una alta diferencia significativa siendo las mejores: RB 73-2223, Mex 56-476, Mex 69-420, RB 73-9735, Mex 53- 473, RB 73-9953, Cp 74-383. Peso promedio de los tallos; Se observó una alta diferencia significativa resultando los mejores genotipos: Mex 56-476, RB 73-2223, Mex 69-420, RB 73-429, Q 96, Mex 68-P23, RB 76-5288, RB 73-9735. Rendimiento agrícola; Estadísticamente se determinó una alta diferencia significativa, las variedades que superan al testigo son: Sp 72-4790, Sp 70- 1284, RB 73-9735. Rendimiento industrial; Se obtuvo diferencia significativa, siendo el mejor genotipo: La Q-96. Rendimiento agro-industrial; No se observaron diferencias significativas entre los cultivares en estudio, de acuerdo a la separación de medias las variedades más destacadas fueron: Sp 72-4790, RB 73- 5220. Respecto a las observaciones fitosanitarias la única variedad que no presento síntomas de las enfermedades señaladas en este estudio es: Sp 72-4790, las variedades más afectadas por el carbón son: RB 74-5672, Cp 72-1210, Cp 70-1527, RB 76-5288, RB 77-320, RB 73-1012, RB 73-9735, L 68-90.

Efecto de diferentes dosis de N.P.K. sobre el rendimiento agroindustrial de la variedad L-68-90 (Saccharum sp hibrido); en caña planta

El presente estudio se realizó de junio de 1992 a marzo de 1993 en El Ingenio Victoria de Julio. Se evaluó el efecto de 13 dosis de N-P-K (0-60-60, 40-60-60, 80-60-60, 120-60-60, 40-0-60, 40-30-60, 80-0-60, 80-30-60, 40-9Ó-60, 120-0-60, 120-30-60, 120-60-0 y 120-60-30), sobre el cultivo de la caña de azúcar (Saccharum sp. híbrido), las cuales se compararon con e1 testigo (0-0-0). Se plantó la variedad L 68-90. El diseño experimental utilizado fue de Bloques Completos al Azar modificado, las variables analizadas fueron: brotación, diámetro, altura, población, rendimiento agrícola, rendimiento industrial y rendimiento agro industrial. Los datos que se obtuvieron se sometieron al análisis de varianza y separación de medías según Tukey a un 5 % de margen de error. En ninguna de las variables evaluadas se observó diferencia estadística significativa. Con respecto a la Germinación el mayor porcentaje fue de 75.33 porciento y correspondió a los tratamientos 120-60-0 y 40-30-60. El mayor diámetro fue de 2.73 cm habiéndose obtenido con el tratamiento 120-60-30. La mayor altura se obtuvo con el tratamiento 40-30-60 y fue de 2.94 cm. La población mayor fue de 231 750 tallos/ha correspondiendo al tratamiento 40-30-60. El mayor rendimiento agrícola se logró con el tratamiento 80-0-60 y fue de 86.23 t/ha. El mayor rendimiento industrial fue de 93.80 kg/t correspondiendo éste al tratamiento 120-0-60. El mayor rendimiento agro-industrial se obtuvo con el tratamiento 80-0-60 siendo éste valor de 7.675 t/ha.

Two-phase flow patterns in a 90 degrees bend at microgravity

Bends are widely used in pipelines carrying single- and two-phase fluids in both ground and space applications. In particular, they play more important role in space applications due to the extreme spatial constraints. In the present study, a set of experimental data of two-phase flow patterns and their transitions in a 90degrees bend with inner diameter of 12.7 mm. and curvature radius of 76.5 mm at microgravity conditions are reported. Gas and liquid superficial velocities are found to range from (1.0 similar to 23.6) m/s for gas and (0.09 similar to 0.5) m/s for liquid, respectively. Three major flow patterns, namely slug, slug-annular transitional, and annular flows, are observed in this study. Focusing on the differences between flow patterns in bends and their counterparts in straight pipes, detailed analyses of their characteristics are made. The transitions between adjoining flow patterns are found to be more or less the same as those in straight pipes, and can be predicted using Weber number models satisfactorily. The reasons for such agreement are carefully examined.

Sapientia, 1968, Vol. XXIII, nº 90 (número completo)

Contenido: La ubicación de la inteligencia / Octavio N. Derisi – La libertad en la filosofía de la integralidad / Alberto Caturelli – Conocimiento y ser / José I. Alcorta – Notas y comentarios -- Bibliografía

Edital nº 90, de 13 de dezembro de 1995 : concurso público para Analista Legislativo : Arquivista e Bibliotecário

Em conformidade com o Edital nº 9/1994, torna pública as notas das prova de Conhecimento Específico II e de Legislação, Língua Portuguesa e Estrangeira, com a respectiva classificação final do concurso público para Analista Legislativo - Arquivista e Bibliotecário.

Análise do sistema partidário brasileiro : a evolução das bancadas parlamentares na década de 90

Examina o sistema partidário brasileiro, considerando a evolução das bancadas parlamentares do PFL, PMDB, PSDB e PT nas eleições de 1990, 1994, 1998 e 2002. Discute a hipótese de que, com a sequência das eleições na década de 1990, o quadro partidário brasileiro estaria começando a apresentar sinais de estabilização.