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81.
Mutations in the fumarate hydratase gene cause hereditary leiomyomatosis and renal cell cancer in families in North America 总被引:10,自引:0,他引:10 下载免费PDF全文
Toro JR Nickerson ML Wei MH Warren MB Glenn GM Turner ML Stewart L Duray P Tourre O Sharma N Choyke P Stratton P Merino M Walther MM Linehan WM Schmidt LS Zbar B 《American journal of human genetics》2003,73(1):95-106
Hereditary leiomyomatosis and renal cell cancer (HLRCC) is an autosomal dominant disorder characterized by smooth-muscle tumors of the skin and uterus and/or renal cancer. Although the identification of germline mutations in the fumarate hydratase (FH) gene in European families supports it as the susceptibility gene for HLRCC, its role in families in North America has not been studied. We screened for germline mutations in FH in 35 families with cutaneous leiomyomas. Sequence analysis revealed mutations in FH in 31 families (89%). Twenty different mutations in FH were identified, of which 18 were novel. Of these 20 mutations, 2 were insertions, 5 were small deletions that caused frameshifts leading to premature truncation of the protein, and 13 were missense mutations. Eleven unrelated families shared a common mutation: R190H. Eighty-one individuals (47 women and 34 men) had cutaneous leiomyomas. Ninety-eight percent (46/47) of women with cutaneous leiomyomas also had uterine leiomyomas. Eighty-nine percent (41/46) of women with cutaneous and uterine leiomyomas had a total hysterectomy, 44% at age < or =30 years. We identified 13 individuals in 5 families with unilateral and solitary renal tumors. Seven individuals from four families had papillary type II renal cell carcinoma, and another individual from one of these families had collecting duct carcinoma of the kidney. The present study shows that mutations in FH are associated with HLRCC in North America. HLRCC is associated with clinically significant uterine fibroids and aggressive renal tumors. The present study also expands the histologic spectrum of renal tumors and FH mutations associated with HLRCC. 相似文献
82.
Aquino AC Jorge JA Terenzi HF Polizeli ML 《Applied microbiology and biotechnology》2003,61(4):323-328
An alpha-amylase produced by Scytalidium thermophilum was purified using DEAE-cellulose and CM-cellulose ion exchange chromatography and Sepharose 6B gel filtration. The purified protein migrated as a single band in 6% PAGE and 7% SDS-PAGE. The estimated molecular mass was 36 kDa (SDS-PAGE) and 49 kDa (Sepharose 6B). Optima of pH and temperature were 6.0 and 60 degrees C, respectively. In the absence of substrate the purified alpha-amylase was stable for 1 h at 50 degrees C and had a half-life of 12 min at 60 degrees C, but was fully stable in the presence of starch. The enzyme was not activated by several metal ions tested, including Ca(2+) (up to 10 mM), but HgCl(2 )and CuCl(2) inhibited its activity. The alpha-amylase produced by S. thermophilum preferentially hydrolyzed starch, and to a lesser extent amylopectin, maltose, amylose and glycogen in that order. The products of starch hydrolysis (up to 6 h of reaction) analyzed by thin layer chromatography, showed oligosaccharides such as maltotrioses, maltotetraoses and maltopentaoses. Maltose and traces of glucose were formed only after 3 h of reaction. These results confirm the character of the enzyme studied to be an alpha-amylase (1,4-alpha-glucan glucanohydrolase). 相似文献
83.
Bessy Gutirrez Luis Osorio María Cristina M. Motta Telervo Huima-Byron Heydeie Erdjument-Bromage Christian Muoz Hernn Sagua Renato A. Mortara Alex Echeverría Jorge E. Araya Jorge Gonzlez 《Parasitology international》2009,58(4):367-374
Three different monoclonal antibodies were produced against Trypanosona cruzi proteasomes. These antibodies were shown to react with a single 27-kDa band on immunoblots of purified proteasomes. Using a 7E5 monoclonal antibody (IgG1) that recognized the α5 subunit of protozoan protease we have studied the intracellular distribution of the T. cruzi 20S proteasome. Contrary to all cell types described to date, T. cruzi 20S proteasome was found not only in the cytoplasm and nucleus but also in the kinetoplast. As revealed by confocal microscopy, the reactivity of monoclonal antibody 7E5 was highly specific for protozoan proteasome because the antibody recognized only the proteasomes from parasites and not those from the mammalian host in T. cruzi infected cells. These findings were confirmed by immunoblots or immunoprecipitations, followed by chymotrypsin-like activity detection in kinetoplasts isolated by differential centrifugation and sucrose density gradients. Proteasome 20S was present in all T. cruzi stages and only slight differences in terms of relative abundance were found. The potential role of the proteasome in kinetoplast remodeling remains to be determined. 相似文献
84.
Hernández-Montiel HL Meléndez-Herrera E Cepeda-Nieto AC Mejía-Viggiano C Larriva-Sahd J Guthrie S Varela-Echavarría A 《Developmental biology》2003,255(1):99-112
We have addressed the control of longitudinal axon pathfinding in the developing hindbrain, including the caudal projections of reticular and raphe neurons. To test potential sources of guidance signals, we assessed axon outgrowth from embryonic rat hindbrain explants cultured in collagen gels at a distance from explants of midbrain-hindbrain boundary (isthmus), caudal hindbrain, or cervical spinal cord. Our results showed that the isthmus inhibited caudally directed axon outgrowth by 80% relative to controls, whereas rostrally directed axon outgrowth was unaffected. Moreover, caudal hindbrain or cervical spinal cord explants did not inhibit caudal axons. Immunohistochemistry for reticular and raphe neuronal markers indicated that the caudal, but not the rostral projections of these neuronal subpopulations were inhibited by isthmic explants. Companion studies in chick embryos showed that, when the hindbrain was surgically separated from the isthmus, caudal reticulospinal axon projections failed to form and that descending pioneer axons of the medial longitudinal fasciculus (MLF) play an important role in the caudal reticulospinal projection. Taken together, these results suggest that diffusible chemorepellent or nonpermissive signals from the isthmus and substrate-anchored signals on the pioneer MLF axons are involved in the caudal direction of reticulospinal projections and might influence other longitudinal axon projections in the brainstem. 相似文献
85.
Rita Rocha Celso M. Teixeira-Duarte João M.P. Jorge João Henrique Morais-Cabral 《Journal of structural biology》2019,205(3):34-43
RCK (regulating conductance of K+) domains are common regulatory domains that control the activity of eukaryotic and prokaryotic K+ channels and transporters. In bacteria these domains play roles in osmoregulation, regulation of turgor and membrane potential and in pH homeostasis. Whole-genome sequencing unveiled RCK gene redundancy, however the biological role of this redundancy is not well understood. In Bacillus subtilis, there are two closely related RCK domain proteins (KtrA and KtrC) that regulate the activity of the Ktr cation channels. KtrA has been well characterized but little is known about KtrC. We have characterized the structural and biochemical proprieties of KtrC and conclude that KtrC binds ATP and ADP, just like KtrA. However, in solution KtrC exist in a dynamic equilibrium between octamers and non-octameric species that is dependent on the bound ligand, with ATP destabilizing the octameric ring relative to ADP. Accordingly, KtrC-ADP crystal structures reveal closed octameric rings similar to those in KtrA, while KtrC-ATP adopts an open assembly with RCK domains forming a super-helix. In addition, both KtrC-ATP and -ADP octamers are stabilized by the signaling molecule cyclic-di-AMP, which binds to KtrC with high affinity. In contrast, c-di-AMP binds with 100-fold lower affinity to KtrA. Despite these differences we show with an E. coli complementation assay that KtrC and KtrA are interchangeable and able to form functional transporters with both KtrB and KtrD. The distinctive properties of KtrC, in particular ligand-dependent assembly/disassembly, suggest that this protein has a specific physiological role that is distinct from KtrA. 相似文献
86.
Galanter JM Fernandez-Lopez JC Gignoux CR Barnholtz-Sloan J Fernandez-Rozadilla C Via M Hidalgo-Miranda A Contreras AV Figueroa LU Raska P Jimenez-Sanchez G Zolezzi IS Torres M Ponte CR Ruiz Y Salas A Nguyen E Eng C Borjas L Zabala W Barreto G González FR Ibarra A Taboada P Porras L Moreno F Bigham A Gutierrez G Brutsaert T León-Velarde F Moore LG Vargas E Cruz M Escobedo J Rodriguez-Santana J Rodriguez-Cintrón W Chapela R Ford JG Bustamante C Seminara D Shriver M Ziv E Burchard EG Haile R 《PLoS genetics》2012,8(3):e1002554
Most individuals throughout the Americas are admixed descendants of Native American, European, and African ancestors. Complex historical factors have resulted in varying proportions of ancestral contributions between individuals within and among ethnic groups. We developed a panel of 446 ancestry informative markers (AIMs) optimized to estimate ancestral proportions in individuals and populations throughout Latin America. We used genome-wide data from 953 individuals from diverse African, European, and Native American populations to select AIMs optimized for each of the three main continental populations that form the basis of modern Latin American populations. We selected markers on the basis of locus-specific branch length to be informative, well distributed throughout the genome, capable of being genotyped on widely available commercial platforms, and applicable throughout the Americas by minimizing within-continent heterogeneity. We then validated the panel in samples from four admixed populations by comparing ancestry estimates based on the AIMs panel to estimates based on genome-wide association study (GWAS) data. The panel provided balanced discriminatory power among the three ancestral populations and accurate estimates of individual ancestry proportions (R2 > 0.9 for ancestral components with significant between-subject variance). Finally, we genotyped samples from 18 populations from Latin America using the AIMs panel and estimated variability in ancestry within and between these populations. This panel and its reference genotype information will be useful resources to explore population history of admixture in Latin America and to correct for the potential effects of population stratification in admixed samples in the region. 相似文献
87.
Bioprocess intensification: a potential aqueous two-phase process for the primary recovery of B-phycoerythrin from Porphyridium cruentum 总被引:1,自引:0,他引:1
Benavides J Rito-Palomares M 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2004,807(1):33-38
A process for the primary recovery of B-phycoerythrin from Porphyridium cruentum exploiting aqueous two-phase systems (ATPS) was developed in order to reduce the number of unit operations and benefit from an increased yield of the protein product. The evaluation of system parameters such as poly(ethylene glycol) (PEG) molecular mass, concentration of PEG as well as salt, system pH and volume ratio was carried out to determine under which conditions the B-phycoerythrin and contaminants concentrate to opposite phases. PEG 1450-phosphate ATPS proved to be suitable for the recovery of B-phycoerythrin because the target protein concentrated to the top phase whilst the protein contaminants and cell debris concentrated in the bottom phase. An extraction ATPS stage comprising volume ratio (Vr) equal to 1.0, PEG 1450 24.9% (w/w), phosphate 12.6% (w/w) and system pH of 8.0 allowed B-phycoerythrin recovery with a purity of 2.9 (estimated as the relation of the 545-280 nm absorbances). The use of ATPS resulted in a primary recovery process that produced a protein purity of 2.9 +/- 0.2 and an overall product yield of 77.0% (w/w). The results reported demonstrated the practical implementation of ATPS for the design of a primary recovery process as a first step for the commercial purification of B-phycoerythrin produced by P. cruentum. 相似文献
88.
Adrians Otero-Arnaiz Silvia Castillo Jorge Weave Guillermo Ibarra-Manríquez 《Biotropica》1999,31(2):243-254
The Coastal Plain of Chiapas (southern Mexico) was formerly covered by large tracts of subhumid tropical forests but is heavily deforested at present. In this region, 15 pastures were selected to characterize species composition of isolated trees, as well as to describe species composition, growth form patterns, and patterns of dispersal units in the vegetation growing under their canopies. The 65 recorded pasture trees belonged to 20 species and 11 families, of which Fabaceae and Moraceae were the most species-rich. Coccoloba barbadensis (Polygonaceae) and Enterolobium cyclocarpum (Fabaceae) were the most abundant and frequent trees in the studied pastures. More than half (55%) of isolated tree species were fleshy-fruited. In the vegetation sampled under pasture trees, 134 species and 45 families were found. Fabaceae and Poaceae had the largest numbers of species. Herbs were the predominant growth form (46.3%), followed by shrubs (23.9%), trees (23.1%), and lianas (2.2%); 6 species could nor be placed in any growth form category. Most species of this flora were fleshy-fruited (43.3%), followed by heavy, gravity-dispersed fruits (17.9%). The analysis of dispersal units by growth form category confirmed the prevalence of fleshy fruits, although their predominance was not so obvious among herb species. Almost half (49.2%) of the flora under pasture trees was typical of secondary vegetation; this pattern was true for herbs but not for most woody species, which were typical of primary vegetation. A numeric classification of the vegetation samples taken under pasture trees produced eight floristic groups, all of which were independent of the specific identity of pasture trees. No significant effect of dispersal unit type of pasture tree on the characteristics of the vegetation growing under them was found. Future attempts to re-create the original forest cover using isolated trees in pastures as regeneration foci should pay more attention to the maintenance of a large specific diversity independently of the dispersal types among these components of tropical landscapes. 相似文献
89.
The heterogeneity of an aspartic acid-containing thermal polymer population has been studied by anion exchange chromatography, amino acid analysis of the resulting fractions and data processing by the principal components method. By using stepwise or continuous gradients of ionic strength both saw-toothed or bell-shaped elution profiles were obtained. This behaviour and the amino acid composition of analyzed fractions suggest a relatively high degree of heterogeneity in the polymer population although less than theoretically expected. This conclusion is compared with the findings reported in proteinoids made by similar procedures. 相似文献
90.