Profile of organ donors in Ceará, northeastern Brazil, from 1998 to 2012.

The use of cadaver donors for transplantation is often the only alternative in the treatment of patients with organ failure. The purpose of this study was to draw a comprehensive profile of solid organ donors in Ceará, northeastern Brazil, from 1998 to 2012. The study was retrospective and based on secondary data regarding sex, age, blood typing, and cause of brain death obtained from the solid organ donor database of the Ceará Transplantation Center covering the period November 1998 to December 2012. During the study period, 976 donors (69% male) were used. Donors were distributed in 4 age groups as follows: 12.9% <18 years, 50.9% 18-40 years, 28.5% 41-60 years, and 7.7% >60 years. The average age was 35 ± 16 years. On the average, female donors were older than male donors (38.4 ± 17 y vs 33.5 ± 16 y; P < .0001). Men were predominant in the age groups 18-40 y (75.3%; P < .0001) and 41-60 y (59.4%; P < .0001). The main causes of brain death were traumatic brain injury (TBI) (56.7%) and stroke (33.1%). The former was more common in men (P < .0001), the latter in women (P < .0001). TBI was caused by traffic accidents (51.4%), of which 50.7% were motorcycle accidents, and urban violence (22.6%), of which 71.2% were associated with firearms. The number of donations increased in the study period (11.2 donors per million population in 1998-2002 to 68.1 in 2008-2012). In Ceará, solid organ donation is on the rise. The predominant donor profile was young men aged 18-40 years with brain death due to TBI caused by traffic accidents and urban violence.

Tumor necrosis factor beta (TNF-ß) NcoI polymorphism is associated with multiple sclerosis in Caucasian patients from Southern Brazil independently from HLA-DRB1.

This study evaluated the association of tumor necrosis factor beta (TNF-ß) NcoI polymorphism with the presence of multiple sclerosis (MS), disability, and HLA-DRB1 alleles in 208 Brazilian MS patients. As controls, 147 healthy individuals were included. The disability was evaluated at baseline and 5-year follow-up using the Expanded Disability Status Scale (EDSS). The TNF-ß genotypes were determined using PCR and restriction fragment length polymorphism and serum TNF-α level was determined using enzyme-linked immunosorbent assay. Among the MS patients, 166 (79.8 %) were white, 39 (18.7 %) were brown, and three (1.4 %) were Asian descents (those were excluded from the further analysis). Among the 205 MS patients, 149 (72.6 %) presented remitting-relapsing MS. The baseline and 5-year follow-up EDSS ranged from 0.0 to 3.0 and from 1.0 to 5.7, respectively. The TNFB2/B2 genotype was associated with the presence of MS among the white patients (p = 0.0443). Brown patients presented higher disability (p = 0.0234) and higher TNF-α levels (p = 0.0463) than white patients. White and brown patients carrying TNFB2/B2 genotype exhibited higher TNF-α levels (p = 0.0354 and p = 0.0309, respectively) than those with other geotypes. Association between TNF-ß NcoI genotypes and HLA-DRB1 alleles was not observed among the MS patients (p > 0.05). Taken together, TNFB2 allele was associated with the presence of MS independently of HLA-DRB1 in white patients and the TNFB2/B2 genotype was associated with increased TNF-α levels in white and brown patients, which could be an important genetic factor candidate for the susceptibility and pathogenesis of MS.

Estudo de biocompatibilidade in vivo e in vitro do extrato hidroalcoólico de Cissus sicyoides L. - Vitaceae em ratos / Study of in vivo and in vitro biocompatibility of Cissus sicyoides L. - Vitaceae hydroalcoholic extracts in rats

Biocompatibilidade é a capacidade de um material exercer funções específicas quando aplicado em contato com tecidos vivos de determinado hospedeiro, sem, contudo, causar danos ou prejuízo ao mesmo. Este trabalho objetivou determinar a biocompatibilidade in vivo e in vitro do extrato hidroalcoólico do Cissus sicyoides L - Vitaceae. Foram utilizados 30 ratos (Rattus novergicus albinus wistar), com idade entre 45 e 90 dias e pesando entre 170 e 260 g. Os animais foram divididos em 3 grupos (A1, A2 e A3) de 6 animais cada para o teste in vivo, os quais foram sacrificados com 2, 4 e 6 dias, respectivamente. Para o teste in vitro, foram utilizados 12 animais para obtenção do índice de aderência e da capacidade fagocítica dos macrófagos de ratos do grupo controle e do grupo experimental. Nos resultados encontrados no teste in vivo, conclui-se que o extrato apresentou-se biocompatível, visto que não provocou alterações significativas no tecido. Já no teste in vitro, o mesmo não se apresentou biocompatível, pois o extrato puro apresentou índice de aderência baixo (7,1) e taxa de fagocitose elevada (35,7), indicando diferença significante quando comparado ao controle. Porém, quando diluído, o extrato se mostrou inócuo, devido ao aumento dos valores do índice de aderência nas concentrações de 1/10 (61,4) e 1/100 (74,3) nos ensaios, as quais não apresentaram diferença significante quando comparadas ao controle. Após a análise dos dados, concluiu-se que a solução diluída do extrato hidroalcoólico do Cissus sicyoides L. não causa danos ou prejuízos. Entretanto, como nem todos os efeitos farmacológicos foram testados no presente trabalho, não se pode inferir automaticamente que ele é biocompatível em todos os casos.

Biocompatibility is the ability of a material to perform specifictasks when applied to living tissues without causing damage or injuries to it. Thus, this study aimed at determining the in vivo and in vitro biocompatibility of Cissus sicyoides L. - Vitaceae hydroalcoholic extracts. A total of 30 rats (Rattus norvegicus Albinus Wistar), with ages ranging from 45 to 90 days and weighing between 170 and 260g were used. The animals were divided into 3 groups (A1, A2 and A3) with 6 animals each, for the in vivo test, which were sacrificed after 2, 4 and 6 days, respectively. For in vitro test, 12 animals were used to obtain the index of adherence and phagocytic ability of macrophages of rats from the control and the experimental groups. In results found for the in vivo test, it was concluded that the extract was biocompatible, whereas no significant changes were observed in the tissue. As to the in vitro test, the extract was not biocompatible, since the pure extract showed a low rate of adherence (7.1) and a high rate of phagocytosis (35.7), indicating a significant difference when compared to the control group. However, when diluted, the extract was shown to be harmless, due to an increase in the values of the adherence index at the following concentrations : 1/10 (61.4) and 1/100 (74.3) in the tests, which showed no significant differences when compared to the control group. After analyzing the data, it was concluded that since the infusion of the plant is a kind of dilution, its use does not cause any harm to the body. A new study is necessary at the moment to possibily demonstrate its effects on the long term.

Successful organ transplantation from donors poisoned with a carbamate insecticide.

Currently, liver transplantation is the only option for patients with end-stage liver disease. In Brazil, the mortality rate on the waiting list is about 25%. Multiple strategies to expand the donor pool are being pursed, however, grafts from poisoned donors are rarely used. This report documents successful liver, kidney and heart transplantations from four female donors who suffered brain death by hypoxia despite cardiopulmonary resuscitation following Aldicarb exposure ([2-methyl-2-(methylthio)propionaldehyde O-(methylcarbamoyl)-oxime]). The success rate of 12 grafts from four donors poisoned by Aldicarb was 91% 6 months after transplantation. Poisoned patients are another pool of organ donors who at present are probably underused by transplantation services. More studies are necessary to confirm the safety for the recipients.

Hypoglycemic effect of Bauhinia cheilandra in rats.

The extract of the methanolic leaves of Bauhinia cheilandra (BC) was tested on glucose loaded and alloxan-induced diabetic rats. In both tests, the methanolic extract at doses of 300, 600, and 900 mg/kg, has shown a statistically significant and considerable hypoglycemic activity.

Analysis of expressed sequence tags from Musa acuminata ssp. burmannicoides, var. Calcutta 4 (AA) leaves submitted to temperature stresses.

In order to discover genes expressed in leaves of Musa acuminata ssp. burmannicoides var. Calcutta 4 (AA), from plants submitted to temperature stress, we produced and characterized two full-length enriched cDNA libraries. Total RNA from plants subjected to temperatures ranging from 5 degrees C to 25 degrees C and from 25 degrees C to 45 degrees C was used to produce a COLD and a HOT cDNA library, respectively. We sequenced 1,440 clones from each library. Following quality analysis and vector trimming, we assembled 2,286 sequences from both libraries into 1,019 putative transcripts, consisting of 217 clusters and 802 singletons, which we denoted Musa acuminata assembled expressed sequence tagged (EST) sequences (MaAES). Of these MaAES, 22.87% showed no matches with existing sequences in public databases. A global analysis of the MaAES data set indicated that 10% of the sequenced cDNAs are present in both cDNA libraries, while 42% and 48% are present only in the COLD or in the HOT libraries, respectively. Annotation of the MaAES data set categorized them into 22 functional classes. Of the 2,286 high-quality sequences, 715 (31.28%) originated from full-length cDNA clones and resulted in a set of 149 genes.

Central antinociceptive effect of a hydroalcoholic extract of Dioclea grandiflora seeds in rodents.

The acute treatment of rats and mice with a hydroalcoholic extract from the seeds of Dioclea grandiflora (EHDg) at doses of 250 and 500 mg/kg, by intraperitoneal or oral administration, produced a significant antinociceptive effect in the tail flick and hot plate tests, an effect which was inhibited by naloxone. EHDg given to mice daily for 30 days at a dose of 500 mg/kg, did not cause any observable toxic effect nor any alteration in the pattern of antinociceptive response by the tail immersion test during the course of this treatment. These results suggest that EHDg has a central antinociceptive action devoid of tolerance effect typical of opioid drugs.

Clinical and histopathological changes of the nasal mucosa induced by occupational exposure to sulphuric acid mists.

AIMS: To assess potential alterations of the nasal mucosa by clinical and histopathological evaluation of workers exposed to sulphuric acid mists at anodising plants, correlating the findings with duration of exposure and sulphuric acid concentrations in the air, and comparing them with a control group. METHODS: Fifty two workers from five plants underwent a clinical evaluation (standard questionnaire, clinical, and ear, nose, and throat examination including nasal endoscopy). For the histopathological study, 20 of the 52 subjects (study group) were randomly selected, as well as 11 unexposed subjects (control group), matched by sex, age, and smoking habits. Nasal biopsy specimens were obtained from the anterior septum mucosa and the anterior curvature of the middle turbinate in each individual. A total of 56 nasal mucosa specimens (37 in the study group and 19 in the control group) were evaluated with regard to normal respiratory epithelium or metaplastic epithelium, atypia or dysplasia, and alterations of the lamina propria. RESULTS: The histopathological study revealed squamous metaplasia in 29 (79%) and atypia in 13 (35%) of the 37 study group samples. No association was found between exposure duration and the clinical and histopathological variables, but a significant association was found between sulphuric acid concentrations higher than 200 micro g/m(3) and pale mucosal patches and ulcerations in the exposed subjects. Logistic regression analysis showed that the exposed subjects had a fivefold risk of developing atypia compared with the unexposed subjects. CONCLUSIONS: Workers exposed to sulphuric acid mists presented with a high incidence of nasal symptoms, and macroscopic and microscopic changes of the nasal mucosa, including squamous atypia and dysplasia. The risk for these histopthological lesions increased with higher sulphuric acid concentrations in the air, revealing an exposure-response relation.

Towards the identification of cassava root protein genes.

The protein population of cassava root layers was characterized by SDS-PAGE and bidimensional polyacrylamide gel electrophoresis. SDS-Page revealed the presence of a protein population in the molecular weight range between 94 and 20 kDa. The expression pattern of these proteins was well-defined within the different layers. Partial protein sequence analyses and preliminary results on the layer-specific expression pattern obtained with Northern analyses are presented.

Toxicological evaluation of the hydro-alcohol extract of the dry leaves of Peumus boldus and boldine in rats.

The hydro-alcohol extract of the dry leaves of Peumus boldus and boldine, showed abortive and teratogenic action and changes in the blood levels of bilirubin, cholesterol, glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and urea in rats. The long term administration of the extract and boldine did not cause histological modification during a period of 90 days.

ACGT and vicilin core sequences in a promoter domain required for seed-specific expression of a 2S storage protein gene are recognized by the opaque-2 regulatory protein.

The expression of Brazil nut storage albumin genes is highly regulated during seed development. Several sequences in the promoter of one of these genes show homologies with the target sites of the maize O2 bZIP regulatory protein. We therefore asked whether the O2 protein would recognize these promoter sequences. We show that the O2 protein binds to three different sequences (F1, F2 and F3). F1 and F3 are hybrid C/G and A/G boxes, respectively, that are homologous to the O2-binding site of a maize alpha-zein gene. F2 is a new O2-binding sequence related to the O2 target sites of the Coix alpha-coxin, the maize b-32 genes and the AP-1 pseudopalindrome. Molecular modelling showed that an Asn and a Ser in the 02 DNA binding domain make different base-specific contacts with each operator. 5' Promoter deletions of the be2S1 gene showed that the domain containing the O2 target sites F1 and F2 is required for detectable reporter gene expression in transgenic tobacco seeds. Moreover, the homologous coix O2 protein was shown to in situ transactivate the promoter region encompassing the three O2-binding sites F1, F2 and F3. Thus, these sites may be in vivo regulatory sequences mediating activation by bZIP regulatory proteins.

Transient expression of GUS and the 2S albumin gene from Brazil nut in peanut (Arachis hypogaea L.) seed explants using particle bombardment.

The effect of parameters involved in the transformation efficiency of peanut (Arachis hypogaea L.) seed tissues by direct gene transfer using a helium inflow particle bombardment device was evaluated. Transient gene expression was affected by both particle and DNA amounts, and was positively correlated with gene copy number, as determined byß-glucuronidase (GUS) activity assays. No influence of plasmid size on GUS gene expression was observed. Transcriptional control of GUS by either the CaMV 35S or the 2S promoter from Brazil nut 2S albumin gene varied with the developmental stage of the seed and was approximately tenfold greater under the influence of the 35S promoter than under the 2S promoter. The gene products of both the Brazil nut methionine-rich 2S albumin and GUS genes under the transcriptional control of the 35S promoter were detected by ELISA assays.

The BRCA1 and 1A1.3B promoters are parallel elements of a genomic duplication at 17q21.

The results of experiments aimed at detecting polymorphisms and mutations in the BRCA1 promoter region as well as comparisons of two published DNA sequences indicated that two similar but distinct copies of this region exist in the human genome. PCR primers specific for amplification of each of the related sequences were developed and new genomic clones corresponding to each of the two promoter regions were isolated from rearrangement-resistant libraries. Sequence analysis of the clones and specific PCR products reveals two similar genomic arrangements of head-to-head genes. The BRCA1 gene is closely apposed to a gene structure that is similar but not identical to 1A1.3B, and the 1A1.3B gene is apposed to a gene structure that has strong similarity to BRCA1 but also significant differences. STS analysis of YAC and P1 clones located in the vicinity of BRCA1 indicates that these similar promoter regions are elements of a direct duplication. New hypotheses for genetic mechanisms that may be involved in breast and ovarian cancer etiology are raised by the identification of this duplicated genetic structure on chromosome 17q.

BRCA1 R841W: a strong candidate for a common mutation with moderate phenotype.

BRCA1 mutations cause increased risk for breast and ovarian cancer, frequently of early onset. Many different mutations occur in BRCA1, including several examples of recurrent mutations, each of which accounts for a significant number of families with heritable cancer predisposition. These common mutations have an etiological role in many breast and ovarian cancer cases and provide the opportunity to examine genotype-phenotype correlations and genotype-environment interactions in individuals with the identical BRCA1 lesion. We report a novel missense change in BRCA1, 2640 C-->T (R841W), found in 3 cases from a subject group of 305 breast and 79 ovarian cancer cases from Orange County, CA. These are consecutive, population-based cases not selected for age or family history. In all three cases, there is a strong family history of breast, ovarian, or other cancers possibly related to a BRCA1 defect and family members showed a high concordance of cancer incidence with the presence of R841W. The age of cancer onset was not always distinct from typical sporadic cases. Testing of a sample of 413 unrelated individuals to examine the hypothesis that R841W might be a rare polymorphism detected one additional instance in a woman with breast cancer diagnosed at age 77 years, and cancer in one parent. R841W is likely to be an etiologically significant lesion with involvement in close to 1% (95% confidence interval of 0-1.7%) of all breast and ovarian cancers in this population.

A corm-specific gene encodes tarin, a major globulin of taro (Colocasia esculenta L. Schott).

A gene encoding a globulin from a major taro (Colocasia esculenta L. Schott) corm protein family, tarin (G1, ca. 28 kDa) was isolated from a lambda Charon 35 library, using a cDNA derived from a highly abundant corm-specific mRNA, as probe. The gene, named tar1, and the corresponding cDNA were characterized and compared. No introns were found. The major transcription start site was determined by primer extension analysis. The gene has an open reading frame (ORF) of 765 bp, and the deduced amino acid sequence indicated a precursor polypeptide of 255 residues that is post-translationally processed into two subunits of about 12.5 kDa each. The deduced protein is 45% homologous to curculin, a sweet-tasting protein found in the fruit pulp of Curculigo latifolia and 40% homologous to a mannose-binding lectin from Galanthus nivalis. Significant similarity was also found at the nucleic acid sequence level with genes encoding lectins from plant species of the Amaryllidaceae and Lilliaceae families.

Schwannoma of the nasal septum. Apropos of 2 cases.

We present two cases of schwannoma of the nasal septum. One of the patients has neurofibromatosis type 2; however, her main complaint was nasal obstruction. Schwannoma deriving from the septum is an extremely rare pathology, both in systemic neurofibromatosis type 2 and as a solitary lesion. There are no characteristic symptoms. Histopathologic examination may be inconclusive so that the definitive diagnosis requires immunohistochemic studies or electron microscopy. Differential diagnosis includes several neurogenic and mesenchymal tumors. CT scan and MRI studies are helpful in evaluating the origin, localization and extension of the lesion. Complete resection of the mass is usually curative, although patients with neurofibromatosis present a higher risk of local recurrence and malignant transformation.

Transgenic plants of ramie (Boehmeria nivea Gaud.) obtained by Agrobacterium mediated transformation.

A regeneration and transformation protocol for ramie (Boehmeria nivea Gaud.) is presented. Regeneration was obtained from leaf discs placed on solid B-5 medium (Gamborg et al. 1968) containing adequate concentrations of auxin and cytokinin. Co-cultivation of leaf discs with Agrobacterium tumefaciens and subsequent regeneration resulted in transgenic plants as shown by Southern blot and analysis of expression of the GUS-marker gene.

Particle bombardment-mediated transient expression of a Brazil nut methionine-rich albumin in bean (Phaseolus vulgaris L.).

Bean (Phaseolus vulgaris L.) mature embryos were transformed using biolistic methods with a plasmid containing 2S albumin and beta-glucuronidase structural sequences, both under the control of the 35S CaMV promoter. We have shown that chimaeric tissues could be obtained and that both structural sequences were expressed to similar levels.

Expression of the 2S albumin from Bertholletia excelsa in Brassica napus.

The methionine rich 2S albumin seed storage protein of Bertholletia excelsa has been expressed in seeds of Brassica napus (rapeseed). A chimeric gene driven by the soybean lectin 5' flanking regions was used to produce a fusion protein consisting of the soybean lectin signal peptide and the propeptide of the Brazil nut 2S albumin. Several transgenic plants were studied at the RNA and protein levels; in each case the chimeric gene was expressed and the protein detected at levels ranging from 0.02% to 0.06% of total protein. Transcriptional studies in a particular transgenic plant show that expression of the gene is tissue specific and developmentally regulated during seed maturation. The endogenous napin genes and the introduced gene are regulated differently, with expression of the chimeric gene paralleling that seen when the soybean lectin gene is expressed in other plant species. Western analysis using antibodies to Brazil nut 2S albumins resulted in the detection of a protein whose size is consistent with correct processing of the precursor.