Recuperation and improvement of chromosome preparations stained with silver nitrate

Both periodic acid and photographic reagents have been used to remove the silver nitrate residues from cytological preparations. We used potassium ferricyanide to remove AgNO3 salts in cases of excessive chromosome impregnation. This method produced partial decolorization, with contrast enhancement. Counterstaining with Giemsa also promoted a better contrast between chromosome arms and NORs, which were preserved and retained a dark color. Application of this procedure to chromosome preparations treated sequentially for CBG banding/AgNO3 staining promoted complete decolorization of C+ regions, leading to "reverse" C-banding, generally with high contrast.

Enhanced expression of Ang-(1-7) during pregnancy

Pregnancy is a physiological condition characterized by a progressive increase of the different components of the renin-angiotensin system (RAS). The physiological consequences of the stimulated RAS in normal pregnancy are incompletely understood, and even less understood is the question of how this system may be altered and contribute to the hypertensive disorders of pregnancy. Findings from our group have provided novel insights into how the RAS may contribute to the physiological condition of pregnancy by showing that pregnancy increases the expression of both the vasodilator heptapeptide of the RAS, angiotensin-(1-7) [Ang-(1-7)], and of a newly cloned angiotensin converting enzyme (ACE) homolog, ACE2, that shows high catalytic efficiency for Ang II metabolism to Ang-(1-7). The discovery of ACE2 adds a new dimension to the complexity of the RAS by providing a new arm that may counter-regulate the activity of the vasoconstrictor component, while amplifying the vasodilator component. The studies reviewed in this article demonstrate that Ang-(1-7) increases in plasma and urine of normal pregnant women. In preeclamptic subjects we showed that plasma Ang-(1-7) was suppressed as compared to the levels found in normal pregnancy. In addition, kidney and urinary levels of Ang-(1-7) were increased in pregnant rats coinciding with the enhanced detection and expression of ACE2. These findings support the concept that in normal pregnancy enhanced ACE2 may counteract the elevation in tissue and circulating Ang II by increasing the rate of conversion to Ang-(1-7). These findings provide a basis for the physiological role of Ang-(1-7) and ACE2 during pregnancy.

Prevalence of deltaF508, G551D, G542X, and R553X mutations among cystic fibrosis patients in the North of Brazil

Cystic fibrosis (CF) is the most common genetic disease among Caucasians and is rare among sub-Saharan Africans. The Brazilian population is not ethnically homogeneous but it is the result of three-way ethnic admixture of Europeans, Africans and Amerindians in varying proportions, depending on the region. In the present study, we investigated 33 patients who had been diagnosed and are currently under treatment for CF at the University Hospital João de Barros Barreto, Belém, Pará State. The molecular analysis for G542X, G551D and R553X mutations was performed by PCR followed by RFLP using BstNI, HincII and MboI, respectively, in polyacrylamide gel eletrophoresis and stained with AgNO3. ThedeltaF508 mutation (a deletion of 3 bp) was only analyzed by polyacrylamide gel electrophoresis and stained with AgNO3. Each sample was analyzed for regions of interest in the CFTR gene using amplified by PCR and specific primers. The deltaF508 and G551D mutations presented frequencies of 22.7 and 3%, respectively. In 74.3% of the remaining patients, none of the mutations investigated was found. The present study characterized in a sample of patients with an established clinical diagnosis of CF (asthma, repeated bronchopneumonia, disorders of nutritional status, etc.) the most frequent mutation ( deltaF508) in the North region of Brazil and is also the first report of the G551D mutation. In spite of the wide spectrum of CF mutations and the heterogeneous ethnic origin of the Amazon population, the molecular diagnosis is a helpful additional tool for the diagnosis and treatment of CF patients.

The validity and 4-year test-retest reliability of the Brazilian version of the Eating Attitudes Test-26

In a cross-sectional study conducted four years ago to assess the validity of the Brazilian version of the Eating Attitudes Test-26 (EAT-26) for the identification of abnormal eating behaviors in a population of young females in Southern Brazil, 56 women presented abnormal eating behavior as indicated by the EAT-26 and the Edinburgh Bulimic Investigation Test. They were each matched for age and neighborhood to two normal controls (N = 112) and were re-assessed four years later with the two screening questionnaires plus the Composite International Diagnostic Interview (CIDI). The EAT results were then compared to diagnoses originating from the CIDI. To evaluate the temporal stability of the two screening questionnaires, a test-retest design was applied to estimate kappa coefficients for individual items. Given the prevalence of eating disorders of 6.2%, the CIDI psychiatry interview was applied to 161 women. Of these, 0.6% exhibited anorexia nervosa and 5.6%, bulimia nervosa (10 positive cases). The validity coefficients of the EAT were: 40% sensitivity, 84% specificity, and 14% positive predictive value. Cronbach's coefficient was 0.75. For each EAT item, the kappa index was not higher than 0.344 and the correlation coefficient was lower than 0.488. We conclude that the EAT-26 exhibited low validity coefficients for sensitivity and positive predictive value, and showed a poor temporal stability. It is reasonable to assume that these results were not influenced by the low prevalence of eating disorders in the community. Thus, the results cast doubts on the ability of the EAT-26 test to identify cases of abnormal eating behaviors in this population.

Lipolysis of emulsion models of triglyceride-rich lipoproteins is altered in male patients with abdominal aorta aneurysm

Disorders of the lipid metabolism may play a role in the genesis of abdominal aorta aneurysm. The present study examined the intravascular catabolism of chylomicrons, the lipoproteins that carry the dietary lipids absorbed by the intestine in the circulation in patients with abdominal aorta aneurysm. Thirteen male patients (72 ± 5 years) with abdominal aorta aneurysm with normal plasma lipid profile and 13 healthy male control subjects (73 ± 5 years) participated in the study. The method of chylomicron-like emulsions was used to evaluate this metabolism. The emulsion labeled with 14C-cholesteryl oleate and ³H-triolein was injected intravenously in both groups. Blood samples were taken at regular intervals over 60 min to determine the decay curves. The fractional clearance rate (FCR) of the radioactive labels was calculated by compartmental analysis. The FCR of the emulsion with ³H-triolein was smaller in the aortic aneurysm patients than in controls (0.025 ± 0.017 vs 0.039 ± 0.019 min-1; P < 0.05), but the FCR of14C-cholesteryl oleate of both groups did not differ. In conclusion, as indicated by the triglyceride FCR, chylomicron lipolysis is diminished in male patients with aortic aneurysm, whereas the remnant removal which is traced by the cholesteryl oleate FCR is not altered. The results suggest that defects in the chylomicron metabolism may represent a risk factor for development of abdominal aortic aneurysm.

Confiabilidade e validade da escala de depressão geriátrica em idosos com doença arterial coronariana

FUNDAMENTO: A prevalência de depressão em portadores de doença arterial coronariana (DAC) é alta. A escala de depressão geriátrica (EDG) é um instrumento amplamente usado para rastrear a depressão em idosos. No Brasil, as propriedades psicométricas da versão curta ainda não foram adequadamente exploradas. OBJETIVO: Avaliar as propriedades psicométricas da versão curta da EDG em portadores de DAC em ambulatório de cardiologia. MÉTODOS: Estudo transversal que avaliou 209 idosos (≥ 65 anos) com DAC utilizando a EDG-15, Cumulative Illness Rating Scale for Geriatrics (CIRS), Brazilian OARS Multidimensional Function Assessment Questionnaire (BOMFAQ) e The Cambridge Examination for Mental Disorders of the Elderly (CAMDEX). A consistência interna da EDG-15 foi calculada pelo KR-20. Uma análise fatorial dessa escala foi conduzida. Escores da EDG-15 foram comparados com os diagnósticos de depressão (DSM-IV) para a validade de critérios. Na análise de validade concorrente, os mesmos escores foram correlacionados com os das escalas de depressão CAMDEX, Miniexame do Estado Mental (MEEM), Cambridge Cognitive Examination (CAMCOG) e BOMFAQ. RESULTADOS: A depressão clínica foi diagnosticada em 35,71% da amostra avaliada de acordo com o DSM-IV. Para o diagnóstico de depressão maior ou distimia, o ponto de corte 5/6 apresentou acurácia moderada (AUROC = 0,84), sensibilidade de 79,92% e especificidade de 78,29%. A consistência interna foi de 0,80. Na análise fatorial, três fatores obtidos explicaram 52,72% da variância total observada. Os escores da EDG-15 correlacionaram-se com os da escala de depressão CAMDEX. CONCLUSÃO: No geral, a EDG-15 apresentou boa confiabilidade e validade (concorrente e de critério). Em settings cardiológicos, seu uso pode auxiliar no rastreamento de quadros depressivos de forma simples e rápida.

Heterotaxy syndrome: a case report

Heterotaxy syndrome is defined as an abnormal arrangement of some organs and vessels in association with dysmorphism. The authors describe the case of a patient with heterotaxy syndrome with poliesplenia incidentally diagnosed during imaging evaluation (computed tomography and small bowel barium study) of unrelated pathological condition.

alfa e b-glucosidases como alvos moleculares para desenvolvimento de fármacos

Glucosidases are involved in key steps in the processing of oligosaccharides by cleaving O-glucose residues. Since they catalyze breaking and transfer reactions of glucosidic groups for the normal growth and development of all the cells, defects or genetic deficiencies in these enzymes are associated with serious disorders of the carbohydrate metabolism. Thus, glucosidases represent important targets to develop inhibitors, owing to their potential activities against viruses, tumoral growth and metastasis, diabetes, Gaucher's disease and other syndromes associated with the lisosomal storage of glucoesphingolipids, and osteoarthritis. This paper presents a description of the biochemical pathways and mechanisms of alpha and beta-glucosidases, and the currently available drugs capable to inhibit these enzymes.

Validação de método analítico por cromatografia líquida de alta eficiência para doseamento do adapaleno em suspensões de nanocápsulas

Adapalene is a retinoid drug often used for disorders of the skin, mainly acne. In this work, an analytical method for the quantification of adapaleno in suspensions of nanocapsules by HPLC was developed and validated. The method was linear in the range of 10-30 µg mL-1, with a good correlation coefficient (r = 0.994). Precision and accuracy analysis showed low relative standard deviation (< 4.6%) and a good recovery percentual (98.2-106.9%). The procedure was specific, linear, precise, exact and robust so that the method can be applied in quantification of adapalene in suspensions of nanocapsules.

Acalasia do esôfago: miotomia com transiluminação

Laparoscopic techniques have provided a new dimension to correct functional disorders of the esophagus, which has stimulated some investigators to recently report the use of laparoscopic cardiomyotomy in the treatment of esophageal achalasia. Now, a new instrument has been added to the current laparoscopic technique to offer a safer and easier method to proceed complete myotomy. After the dissection of the esophagogastric junction, a special catheter is introduced reaching the stomach. lt has an illuminated 10 cm extremity connected to a light source. lts withdrawal allows to visualize every muscle circular fiber by transillumination withan improved view provide by the laparoscopic optic system lens. This condition modifies the operative surgeon s attitude offering a better controlled situation over the procedure. The use of transillumination o fthe esophagogastric junction provides a good identification of the mucosa e submucosa avoiding the risk of esophageal perforation. It also helps to perform a complete myotomy preventing the ocurrence of persistent disphagia in the postoperative period. Cardiomyotomy with parcial fundoplication is possible by videolaparoscopic approach, now made easier with transillumination. This technique is safe and the functional results are similar to those observed in the literature for conventional open procedures, with the obvious advantages of the minimally invasive approach.

Derivação biliopancreática com preservação gástrica videolaparoscópica: sistematização técnica

BACKGROUND: The authors describe a videolaparoscopic technical variation of biliopancreatic diversion, and its rationale, as well as its preliminary results.They operated on 12 female patients, age between 26 and 49 years, BMI range 47,4 and 59,5kg/m². There were no intraoperative complications; operative time ranged from 2 to 4,5 hours; discharge in the 3rd. postoperative day. Liquid dietetic recommendations in the early period, and normal diet after the first week. One patient was readmmitted after 1 week with abdominal pain because of excessive ingestion of food; there was deep venous trombosis in one patient. Follow-up showed loss of excess weight of 27,4% after four months. Initial results of the described operation sugests it may be a good alternative to morbid obesity surgical treatment; it is strongly required a multidisciplinar pre and postoperative treatment program.

Serum cytokine levels in autoimmune and non-autoimmune hyperthyroid states

Although the role of interleukin-2 (IL-2) and interferon gamma (gIFN) is still poorly understood in hyperthyroid diseases, it is reasonable to assume that these cytokines may be present at higher levels in Graves' disease (GD) than in other primarily non-autoimmune thyroid diseases. In order to look for an easy method to distinguish GD from primarily non-autoimmune causes of hyperthyroidism, we compared 13 healthy individuals with 21 treated and untreated hyperthyroid GD patients and with 19 patients with hyperthyroidism due to other etiologies: 7 cases of multinodular goiter, 5 cases of excessive hormone replacement and 7 cases of amiodarone-associated hyperthyroidism. All patients presented low TSH levels and a dubious clinical thyroid state. We found a good correlation between TSH and serum IL-2 levels (r = 0.56; P<0.01). Serum IL-2 (P<0.01) and gIFN (P<0.01) levels were lower in the hyperthyroid group of patients than in control subjects, suggesting a depressed TH1 pattern in the T-cell subset of hyperthyroid patients. GD had normal IL-2 levels, while patients with other forms of thyrotoxicosis presented decreased IL-2 levels (P<0.05). There was no difference between treated and untreated GD patients. We suggest that the direct measurement of serum IL-2 level may help to confirm hyperthyroidism caused by GD.

Protein defects in neuromuscular diseases

Muscular dystrophies are a heterogeneous group of genetically determined progressive disorders of the muscle with a primary or predominant involvement of the pelvic or shoulder girdle musculature. The clinical course is highly variable, ranging from severe congenital forms with rapid progression to milder forms with later onset and a slower course. In recent years, several proteins from the sarcolemmal muscle membrane (dystrophin, sarcoglycans, dysferlin, caveolin-3), from the extracellular matrix (alpha2-laminin, collagen VI), from the sarcomere (telethonin, myotilin, titin, nebulin), from the muscle cytosol (calpain 3, TRIM32), from the nucleus (emerin, lamin A/C, survival motor neuron protein), and from the glycosylation pathway (fukutin, fukutin-related protein) have been identified. Mutations in their respective genes are responsible for different forms of neuromuscular diseases. Protein analysis using Western blotting or immunohistochemistry with specific antibodies is of the utmost importance for the differential diagnosis and elucidation of the physiopathology of each genetic disorder involved. Recent molecular studies have shown clinical inter- and intra-familial variability in several genetic disorders highlighting the importance of other factors in determining phenotypic expression and the role of possible modifying genes and protein interactions. Developmental studies can help elucidate the mechanism of normal muscle formation and thus muscle regeneration. In the last fifteen years, our research has focused on muscle protein expression, localization and possible interactions in patients affected by different forms of muscular dystrophies. The main objective of this review is to summarize the most recent findings in the field and our own contribution.

Cell therapy in dilated cardiomyopathy: from animal models to clinical trials

Dilated cardiomyopathy can be the end-stage form and common denominator of several cardiac disorders of known cause, such as hypertensive, ischemic, diabetic and Chagasic diseases. However, some individuals have clinical findings, such as an increase in ventricular chamber size and impaired contractility (classical manifestations of dilated cardiomyopathy) even in the absence of a diagnosed primary disease. In these patients, dilated cardiomyopathy is classified as idiopathic since its etiology is obscure. Nevertheless, regardless of all of the advances in medical, pharmacological and surgical procedures, the fate of patients with dilated cardiomyopathy (of idiopathic or of any other known cause) is linked to arrhythmic episodes, severe congestive heart failure and an increased risk of sudden cardiac death. In this review, we will summarize present data on the use of cell therapies in animal models of dilated cardiomyopathies and will discuss the few clinical trials that have been published so far involving patients affected by this disease. The animal models discussed here include those in which the cardiomyopathy is produced by genetic manipulation and those in which disease is induced by chemical or infectious agents. The specific model used clearly creates restrictions to translation of the proposed cell therapy to clinical practice, insofar as most of the clinical trials performed to date with cell therapy have used autologous cells. Thus, translation of genetic models of dilated cardiomyopathy may have to wait until the use of allogeneic cells becomes more widespread in clinical trials of cell therapies for cardiac diseases.

Renegociação da dívida pública interna em condições de sobreendividamento

Renegotiation of the public debt in conditions of excessive debt. This paper studies the debt overhang models and maturity management models, and analyses both theoretically and historically the debt renegotiations which the final outcome is a lower public debt burden. From the theoretical view the renegotiation plans can be Pareto improving, both creditors and debtors can be better-off, and the value of the new debt will price the new reputation and debtors' willingness to pay. From the historical point of view, funded debts were the instrument that debtors used to improve reputation.