Identification of an exported heat shock protein 70 in Plasmodium falciparum

Host cell remodelling is a hallmark of malaria pathogenesis. It involves protein folding, unfolding and trafficking events and thus participation of chaperones such as Hsp70s and Hsp40s is well speculated. Until recently, only Hsp40s were thought to be the sole representative of the parasite chaperones in the exportome. However, based on the re-annotated Plasmodium falciparum genome sequence, a putative candidate for exported Hsp70 has been reported, which otherwise was known to be a pseudogene. We raised a specific antiserum against a C-terminal peptide uniquely present in PfHsp70-x. Immunoblotting and immunofluorescence-based approaches in combination with sub-cellular fractionation by saponin and streptolysin-O have been taken to determine the expression and localization of PfHsp70-x in infected erythrocyte. The re-annotated sequence of PfHsp70-x reveals it to be a functional protein with an endoplasmic reticulum signal peptide. It gets maximally expressed at the schizont stage of intra-erythrocytic life cycle. Majority of the protein localizes to the parasitophorous vacuole and some of it gets exported to the erythrocyte compartment where it associates with Maurer's clefts. The identification of an exported parasite Hsp70 chaperone presents us with the fact that the parasite has evolved customized chaperones which might be playing crucial roles in aspects of trafficking and host cell remodelling.

Insertion Reactions of Six-Membered Cyclopalladated N,N `,N `'-Triarylguanidine, Pd{kappa(2)(C,N)-C6H3Me-3(NHC(NHAr)(=NAr))-2)(mu-Br)](2) (Ar=2-MeC6H4) with PhCC C-C(O)OR (R = Me and Et): A Gateway to Second Orthopalladation through Novel Rearrangements

The reaction of Pd{kappa(2)(C,N)-C6H3Me-3-(NHC(NHAr)(=NAr))-2}(mu-Br)](2) (Ar = 2-MeC6H4; 1) with 4 equiv of PhC C-C(O)OMe in CH2Cl2 afforded Pd{kappa(2)(C,N)-C(Ph)=C(C(O)OMe)C(Ph)=C(C(O)-OMe)C6H3Me-3(N=C(NH Ar)(2))-2}Br] (Ar = 2-MeC6H4; 2) in 70% yield, and the aforementioned reaction carried out with 10 equiv of PhC C-C(O)OR (R = Me, and Et) afforded an admixture of two regioisomers of Pd{kappa(3)(N,C,O)-O=C(OR)-C5Ph3(C(O)OR)C(C(O)OR)C6H3Me-3(N=C(NHAr)( 2))- 2}Br] (Ar = 2-MeC6H4; R = Me (3a/3b), Et (4a/4b)) in 80 and 87% yields, respectively. In one attempt, the minor regioisomer, 4b, was isolated from the mixture in 6% yield by fractional crystallization. Palladacycles 3a/3b and 4a/4b, upon stirring in CH2Cl2/MeCN (1/1, v/v) mixture at ambient condition for S days, afforded Pd{eta(3)-allyl,(KN)-N-1)-C-5(C(O)OR)(2)Ph3C-(C(O)OR)C6H3Me-3(N=C(NH Ar)(2))(-2)}Br] (Ar = 2-MeC6H4; R = Me (5a/5b), Et (6a/6b)) in 94 and 93% yields, respectively. Palladacycles 3a/3b and 4a/4b, upon reaction with AgOTf in CH2CH2/Me2C(O) (1/1, v/v) mixture at ambient temperature for 15 min, afforded Pd{kappa(3)(N,C,O)-O=C(OR)C5Ph3(C(O)OR)C(C(O)OR)C6H3Me-3(N=C(NHAr)(2 ))-2}(OTf)] (Ar = 2-MeC6H4; R = Me (7a/7b), Et (8a/8b)) in 79 and 77% yields, respectively. Palladacycles 7a/7b and 8a/ 8b, upon reflux in PhC1 separately for 6 h, or palladacycles 5a/5b and 6a/6b, upon treatment with AgOTf in CH2Cl2/Me2C(O) (7/3, v/v) mixture for 15 min, afforded Pd{(eta(2)-Ph)C5Ph2(C(O)OR)kappa(2)(C,N)-C(C(O)OR)C6H3Me-3(N=C(NHAr) (2))-2}(OTf)] (Ar = 2-MeC6H4; R = Me (9a/9h), Et (10a/10b)) in >= 87% yields. Palladacycles 9a/9b, upon stirring in MeCN in the presence of excess NaOAc followed by crystallization of the reaction mixture in the same solvent, afforded Pd{kappa(3)(N,C,C)-(C6H4)C5Ph2(C(O)OMe)(2)C(C(O)OMe)(2)C6H3Me-3(N=C( NHAr)(2))-2}(NCMe)] (Ar = 2-MeC6H4; 11a/11b) in 82% yield. The new palladacycles were characterized by analytical, IR, and NMR (H-1 and C-13) spectroscopic techniques, and the molecular structures of 2, 3a, 4a, 4b, 5a, 6a, 7a, 9a, 10a, and 11a-d(3) were determined by single crystal X-ray diffraction. The frameworks in the aforementioned palladacycles, except that present in 2, are unprecedented. Plausible pathways for the formation of new palladacycles and the influence of the guanidine unit in 1, substituents in alkynes, reaction conditions, and electrophilicity of the bromide and the triflate upon the frameworks of the insertion products have been discussed.

Mono- and Dialkyne Insertion Reactions of Cyclopalladated N,N `,N `'-Triarylguanidines kappa(2)(C,N)Pd(mu-Br)](2) and cis-/trans-kappa(2)(C,N)Pd(Lewis Base)Br]. Scaffolds for Enlarged, Rearranged, and Zwitterionic Palladacycles through Ring Contraction cum Amine-Imine Tautomerization

Insertion reactions of six-membered cyclopalladated N,N',N''-triarylguanidines, kappa(2)(C,N)Pd(mu-Br)](2) with various alkynes in CH2Cl2 under ambient conditions afforded diinserted eight-membered palladacycles, (kappa(2)(C,N):eta(2)(C=C)-PdBr] (1-11), in high yield (76-96%), while insertion reactions of six-membered cyclopalladated N,N',N''-triarylguanidines, kappa(2)(C,N)Pd(Lewis base)Br] (VI-XI), with various alkynes under the aforementioned conditions afforded monoinserted six-membered palladacycles, kappa(2)(C,N)-Pd(Lewis base)Br] (12-21), in high yield (81-91%) except for 14 (23%). The insertion reaction of VI with 2 equiv of dimethyl acetylenedicarboxylate (DMAD) and the insertion reaction of 12 with 1 equiv of DMAD in CH2Cl2 under ambient conditions resulted in the formation of a diinserted zwitterionic five-membered palladacycle, kappa(2)(C,C)Pd(2,6-lutidine)Br] (22), in 76% and 70% yields, respectively. Palladacycle 22 upon reaction with AgOTf in wet MeCN afforded the ionic palladacycle kappa(2)(C,C)Pd(2,6-lutidine)(H2O)]OTf] (23) in 78% yield. The ring size of the ``kappa(2)(C,N)Pd]'' unit in the structurally characterized diinserted palladacycles (1 center dot 2CH(2)Cl(2)center dot H2O, 2, 5, and 7), and monoinserted palladacycles (17, 18, and 20 center dot C7H8 H2O) is smaller than that anticipated for mono- and diinserted palladacycles, and this feature is mainly ascribed to the proclivity of III-XI to undergo ring contraction cum amine-imine tautomerization upon alkyne insertion. Palladacycle 22 represents the first diinserted product obtained in alkyne insertion reactions of kappa(2)(C,N)Pd(Lewis base)X] type palladarycles. The molecular structure of 22 center dot H2O determined by X-ray diffraction indicates that the positive charge on the guanidinium moiety is balanced by the negative charge on the palladium atom and thus represents the first structurally characterized zwitterionic palladacycle to be reported in alkyne insertion chemistry. Plausible mechanisms of formation of 12-21 and 22 have been outlined. The presence of more than one species in solution for some of the palladacycles in the series 1-7 and 12-21 was explained by invoking the C-N single-bond rotation of the CN3 unit of the guanidine moiety, while this process in conjunction with Pd-N(lutidine) bond rotation was invoked to explain the presence of four isomers of 15, as studied with the aid of variable-concentration H-1 NMR experiments carried out for 14 and 15.

Structural insights into N-terminal to C-terminal interactions and implications for thermostability of a (beta/alpha)(8)-triosephosphate isomerase barrel enzyme

Although several factors have been suggested to contribute to thermostability, the stabilization strategies used by proteins are still enigmatic. Studies on a recombinant xylanase from Bacilllus sp. NG-27 (RBSX), which has the ubiquitous (beta/alpha)(8)-triosephosphate isomerase barrel fold, showed that just a single mutation, V1L, although not located in any secondary structural element, markedly enhanced the stability from 70 degrees C to 75 degrees C without loss of catalytic activity. Conversely, the V1A mutation at the same position decreased the stability of the enzyme from 70 degrees C to 68 degrees C. To gain structural insights into how a single extreme N-terminus mutation can markedly influence the thermostability of the enzyme, we determined the crystal structure of RBSX and the two mutants. On the basis of computational analysis of their crystal structures, including residue interaction networks, we established a link between N-terminal to C-terminal contacts and RBSX thermostability. Our study reveals that augmenting N-terminal to C-terminal noncovalent interactions is associated with enhancement of the stability of the enzyme. In addition, we discuss several lines of evidence supporting a connection between N-terminal to C-terminal noncovalent interactions and protein stability in different proteins. We propose that the strategy of mutations at the termini could be exploited with a view to modulate stability without compromising enzymatic activity, or in general, protein function in diverse folds where N and C termini are in close proximity. Database The coordinates of RBSX, V1A and V1L have been deposited in the PDB database under the accession numbers 4QCE, 4QCF, and 4QDM, respectively

Reply to Discussion on ``Bearing capacity of circular footings over rock mass by using axisymmetric quasi lower bound finite element limit analysis'' Comput. Geotech. 70 (2015) 138-149]

A discussion has been provided for the comments raised by the discusser (Clausen, 2015)1] on the article recently published by the authors (Chakraborty and Kumar, 2015). The effect of exponent alpha for values of GSI approximately smaller than 30 becomes more critical. On the other hand, for greater values of GSI, the results obtained by the authors earlier remain primarily independent of alpha and can be easily used. (C) 2015 Elsevier Ltd. All rights reserved.

Comportamiento agro-industrial de catorce variedades de caña de azúcar (Saccharum sp hibrido L.) en comparación con las variedades Ja 60-5 y C 87-51

El presente trabajo se llevó a cabo en el ciclo 92-93 en el Ingenio Victoria de Julio, donde se evaluaron 14 variedades de caña de azúcar en comparación con las variedades Ja 60 - 5 y C 87 - 51 para determinar el comportamiento agro-industrial de estos cultivares y seleccionar los genotipos más sobresalientes. El experimento fue establecido el 25 de enero de 1992, en un suelo arcilloso de la serie Chilamatillo, la cosecha se realizó a los 11 meses de edad del cultivo, el 16 de Dic 1992. El diseño utilizado fue de bloques completos al azar con 5 repeticiones, las variables estudiadas fueron: Germinación, población, altura de tallos, floración, diámetro de tallos, peso promedio de los tallos, rendimiento agrícola, rendimiento industrial y rendimiento agro-industrial. Los datos fueron sometidos a un análisis de varianza y prueba de tukey al 5% de margen de error. Los resultados obtenidos fueron los siguientes: Germinación; siendo las mejores RB 73-1012, CP 70-321 y Q 96. Población;las mejores numéricamente resultaron Méx 68-P 23,SP 70-1284 y SP 72-4790. Longitud de tallo; las mejores variedades fueron la SP 72-4790, Q 96, RB 73-1012 y RB 73-5220. Diámetro de los tallos; obtuvo el mejor resultado la variedad Méx 69-420 y en segundo lugar la Ja 60-5. Peso promedio de 100 tallos. ; presentando el mejor resultado el cultivar Méx 69-420 seguido de la RB 73-5220. Floración; todas las variedades florecieron a excepción del testigo C 87-51. Rendimiento agrícola; obtuvo el mejor resultado la variedad Méx 68-P 23 ocupando el segundo lugar SP 72-4790; Rendimiento industrial; entre las variedades sobresalieron numéricamente la Q 96 seguido de la C 87-51 y la CP 72-1210. Rendimiento Agro-Industrial; de acuerdo a los resultados de rangos múltiples la variedad Q 96 presenta un rendimiento estadísticamente superior a la variedad testigo Ja 60-5, superando numéricamente al resto de variedades. Las variedades que presentaron afectaciones por enfermedades fueron : RB 76-5288, SP 70-1284, SP 72-4790, CP 72-1210, SP 71-61801 CP 72-2086, Méx 69-420, Méx 68-P23, Ja 60-5 y e 87-51.

Evaluación de cinco fungicidas para el manejo de enfermedades foliares y su rentabilidad en el cultivo de tomate (Lycopersicom esculentum mill) c.v. butte, Sébaco, Matagalpa, Nicaragua

El presente trabajo se realizó en la Finca Surco Muerto, Municipio de Sébaco–Matagalpa en el período comprendido de Julio a octubre del 2004, con la finalidad de evaluar diferentes productos fungicidas sistémicos (Phyton 0.5 L. ha-1, Benomil 0.5 kg ha-1 y Curzate 2 kg ha-1) y preventivos (Mancozeb 2 kg ha-1 y Clorotalonil 2 L. ha-1) en el manejo de enfermedades foliares en tomate. El diseño establecido fué el de Bloques Completos al Azar (BCA),con siete tratamientos y cuatro repeticiones. Los resultados indican que el efecto de los tratamientos evaluados sobre el control de Alternaria solani enlas primeras fechas no demuestran diferencia estadística, hasta los 62 días después del trasplante (ddt), donde Clorotalonil se comportó como el mejor tratamiento en protección al follaje.Para la variable severidad de Xanthomonas campestris en follaje los resultados indican, que es a partir de los 78 ddt donde los tratamientos demuestran diferencia estadística, comportándose Phyton como el mejor tratamiento para el manejo de dicha enfermedad.El cultivo también fué fuerte mente afectado por Mosca blanca(Bemisia tabaciGenn.) lo que repercutió en porcentajes de severidad de virosis muy altos(92%),enmascarando un mejor efecto que pudieron haber tenido los tratamientos evaluados. Enrelación a las variables de rendimiento analizadas por contrastes ortogonales, para la variable peso de frutos buenos, el análisis no encontró diferencias estadísticas entre los grupos evaluados.Para la variable peso de frutos afectados por Alternaria el análisis detectó diferencias estadísticas entre los grupos evaluados donde el grupo de los Preventivos (Clorotalonil y Mancozeb) ejercieron mejor control para dicho patógeno por haberse obtenido con ellos los más bajos rendimientos afectados con 40.85 kg ha-1. En la variable peso de frutos afectados por Xanthomonas Campestris pvvesicatoria el análisis no encontró diferencias estadísticas entre los grupos comparados.Para el rendimiento real el análisis encontró diferencias estadísticas, donde demuestra que son los tratamientos preventivos (Mancozeb y Clorotalonil) los que ejercieron el mejor control con el más alto rendimiento 5658.56 kg ha-1.Los resultados del análisis económico indican que el tratamiento rentable es Mancozeb, por obtenerse con el una tasa de retorno marginal de 960.25%. En condiciones de bajo rendimiento es Alternado (Curzate + Clorotalonil +Mancozeb) el tratamiento rentable por obtenerse con el una TRM de 246.95%. Al realizar el análisis de sensibilidad los resultados demuestran que la aplicación de Mancozeb es justificable; aun cuando el precio del tomate disminuye en un 70%, de su precio original,ya que con precios bajos se obtiene una TRM de 1.98%.

Influencia de diferentes métodos de control de malezas en soya (Glycine max (L) (Merr) c.v. Cristalina inoculada y sin inoculación

Se realizó un ensayo en el Centro Experimental del Algodón (CAL) Posoltega en la época de postrera de 1988 en el cultivo de soya (Glyceni Max) L Merr) C. V: Cristalina, con el objetivo de determinar la influencia de diferentes métodos de control de malezas sobre el comportamiento de ls cenosis, el crecimiento y el rendimiento de la soya y determinar el efecto de la inoculación sobre cenosis, el crecimiento y rendimiento del cultivo. Se utilizó un diseño de parcelas divididas en bloques al Azar con cuatro réplicas utilizando soya inoculada y sin inocular y tres métodos de control de malezas; La abundancia y la dominancia de malezas fue mayor en soya inoculada y en el control químico con remesaren (Flex 0.70 lts/ha P.C.) ; la diversidad fue similar en los dos tipos de soya; pero el control por periodo crítico (b2) aumento la diversidad a la cosecha superando hasta en 31.2% al control químico (B1). C. rotundus fue la maleza más abundante en todo el ciclo del cultivo. La altura de planta y el número de planta /m2, fueron en soya sin inoculante (a2) con diferencias significativas, para el • número de ramas /plantas • de vainas/planta, peso de 1000 semillas (g), rendimientos (kg/ha), peso seco de la paja (kg/ha). Diámetro del tallo (mm) y altura de inserción de la primera vaina para los dos tipos de soya no se presentaron diferencia estadísticas: El control químico (b1) fue superado en la altura de la planta (cm), pero seca de la paja (Kg/ha), diámetro del tallo (mm), • de vainas/planta y peso de 1000 semillas (g) por los otros dos métodos (b3 y b2), con diferencias significativas. No se encuentra ron diferencias significativas para el • de nódulos/planta y peso seco de nódulos/planta en los niveles de soya y en los diferentes métodos químicos(b1) fue superado significativamente en • de nódulos/plantas por el control en periodo critico (b2). Influencia de diferentes Métodos de control de malezas en soya (Glycine max L Merr) C.V. Cristalina, inoculada y sin inoculante.

P.v.b. virgillii maronis : bucolicorum, eclogaex., georgicorum, libri iii., aeneidos, libri xii. et in ea, mauri servii honorati : grammatici comentarii, ex antiquiss : exemplaribus longe meliores et auctiores : ex bibliotheca petri danielis i.c. : accessit fabij planciadis fulgentij liber de continentia virgiliana, auctior e mss. codd. item iunij philargyrij commentariolus in bucolica & georgica virgilij. : cum certissimo et copiosissimo indice

As Bucólicas são constituídas de dez éclogas, consideradas como traduzidas ou imitadas de Teócrito. As Geórgias são um poema didático no qual se acentua a predileção de Virgílio pela vida campestre. A Eneida é uma utopia política que o poeta não conseguiu acabar e foi publicada contra a sua vontade expressa.

Ana C.: as tramas da consagração

O trabalho tem por objetivo percorrer as principais linhas de força discursivas que se articularam no processo de consagração da poeta Ana Cristina Cesar, e que configuraram a sua imagem mais conhecida na mídia cultural e na academia. Na Introdução, define-se o conceito de autor que operou no desenvolvimento da pesquisa, e a sua relação com a assinatura Ana C.. No primeiro capítulo, é analisada a colocação de Ana Cristina no campo cultural e poético da década de 70; a sua posição dupla de convívio e diferenciação em relação aos poetas da chamada geração marginal. Percorrem-se as leituras críticas que dessa colocação se fizeram, analisando como crítica e objeto de estudo foram se conformando simultaneamente. No segundo capítulo, são definidas as narrativas dominantes que se fizeram dos textos de Ana C. depois da sua morte, contrapondo a imagem oficial que se desprende das edições póstumas, que criam uma imagem mítica de Ana C., a leituras contrárias. No terceiro capítulo, percorre-se a fortuna crítica, geralmente ligada à academia, e textos literários que tomaram a figura de Ana C. como problemática ou tema da escrita, tentando fazer uma avaliação em relação à reafirmação ou a mobilização da imagem consagrada que esses textos propõem

Flavio Josefo e o paradigma de circularidade cultural entre as comunidades judaicas e a sociedade romana na Urbs do século I d.C

O paradigma de circularidade cultural entre a comunidade judaica e a sociedade romana foi construído pela historiografia através da análise do contato sociocultural e embates entre romanos e judeus que, ao longo da história, ocuparam o mesmo espaço em diversas regiões anexadas ao Império Romano. As relações de poder estabelecidas entre Roma e Jerusalém, após a ocupação da Judéia, apontam para uma hierarquização nas relações sociais, culturais e políticas entre romanos e judeus. O conceito de circularidade cultural de Carlo Ginzburg nos permite, a partir da trajetória de Flávio Josefo, identificar a dualidade no mundo social de Josefo, na qual, de um lado estavam as culturas dominantes (sociedade romana) e, do outro, as culturas subalternas (comunidades judaicas) que, apesar da marcação das diferenças, se influenciavam reciprocamente.

中国4种蝙蝠的G-带和C-带

首次报道了中国4种蝙蝠的G-带和C-带核型。大长舌果蝠(Eonycteris spelaea)二倍染色体数目(2n)为36,常染色体臂数(FN)为56;马来假吸血蝠(Megaderma spasma)2n=38,FN=70;黑髯墓蝠(Taphozous melanopogon)2n=42,FN=64;皱唇蝠(Chaerephon plicata)2n=48,FN=54。通过C-带显示,除着丝粒异染色质外,在皱唇蝠的许多染色体臂内和马来假吸血蝠染色体的端粒处也有较多的插入异染色质,大长舌果蝠的基因组中既有臂内异染色质也有端粒异染色质。

Exploratory fishing experiments in Hirakud reservoir - Orissa State (1967-70)

Suitable areas for fishing have been located in the middle and upper regions of the Mahanadi course of Hirakud reservoir. In the former the suitable period is during summer and beginning of monsoons, and in the latter in summer and winter months. The fishery of the reservoir is contributed by four species namely S. silondia, L. fimbriatus, C. mirgala and C. catla.

The first study on the effects of microcystin-RR on gene expression profiles of antioxidant enzymes and heat shock protein-70 in Synechocystis sp PCC6803

Microcystins are heptapeptide toxins produced by cyanobacteria. Microcystin-RR(MC-RR) is a common variant among the 80 variants identified so far. There have been many investigations documenting the toxic effects of microcystins on animals and higher plants, but little is known on the toxic effects of microcystins on algae, especially at molecular level. We studied the effects of MC-RR on gene expression profile of a few antioxidant enzymes and heat shock protein-70 (Hsp70) in Synechocystis sp. PCC6803. After two days post-exposure, a high dose toxin (5 mg/l, about 4.8 x 10(-3) mM) significantly increased expression levels of the genes gpx1, sodB, katG, acnB, gamma-TMTand dnaK2, while a relatively low dose toxin (1 mg/l, about 9.63 x 10(-4) mM) induced a moderate and slow increase of gene expression. Our results indicate that MC-RR could induce the oxidative stress in Synechocystis sp. PCC6803 and the increase in gene expression of antioxidant enzymes and Hsp70 might protect the organism from the oxidative damage. in addition, cell aggregation was observed during the early period of exposure, which might be a specific oxidative stress reaction to MC-RR. (C) 2008 Elsevier Ltd. All rights reserved.

Rickettsia-like organism infection in a freshwater cultured fish Ophiocephalus argus C. in China

From 2001 to 2002, a new and emergent infectious disease of Ophiocephalus argus occurred in a fishery in Hubei Province, China, with an incidence of 60% similar to 70% and a mortality as high as 100 %. The diseased fish showed an enlarged abdomen, the millet-like nodules in internal organs, and the swollen kidney which was composed of 5 similar to 10 sarcoma-like bodies in cream or gray-white colour or ulcerated into beandregs-like substance. Light microscopic observation revealed the basophilic or acidphilic inclusions in cytoplasm of the cells and the granulomas, a diffusive chronic inflammation in internal organs. Further analysis under an electron microscope indicated that the intracytoplasmic inclusions were rickettsia-like organisms (RLOs) that are either spherical or coccoid, with variable size, ranging from 0.5 similar to 1.5 mum in diameter, and enclosed within membrane-bound cytoplasmic vacuoles. RLO had a central nucleoid region with some fine filamentous structures and an electron-dense granule. Its cytoplasm contained abundant ribosomal bodies. Occasionally, RLO appeared to be divided by binary fission. RLOs were also observed in the homogenized tissue of infected fish. The results suggested that the death of cultured O. argus was caused by RLO infection.