Caracterização anatômica, histoquímica e de classes de metabólitos secundários de folhas de Guzmania lingulata (L. ) Mez / Anatomical, histochemical and class characterization of secondary metabolites in leaves of Guzmania lingulata (L. ) Mez

Na medicina natural pouco se sabe sobre a importância curativa da maior parte das espécies pertencentes à Bromeliaceae. Tal fato evidencia a necessidade da realização de investigações para a comprovação de possíveis potenciais fitoterápicos em espécies dessa família. Guzmania lingulata é caracterizada como sendo uma das espécies de bromélia mais comercializada e conhecida popularmente, a qual é amplamente propagada no Brasil. O presente trabalho objetivou a caracterização anatômica e a identificação de classes de metabólitos secundários presentes em G. lingulata, por meio de análises anatômicas, histoquímicas e químicas. As folhas foram fixadas em FAA 50 e transferidas para etanol 50%. Para o estudo anatômico, secções transversais da porção mediana das lâminas foliares foram obtidas à mão livre e submetidas ao processo de dupla coloração com safranina e azul de astra, sendo, logo após, montadas em gelatina glicerinada. Para a triagem das classes de metabólitos secundários foram coletadas 10 folhas frescas que passaram pelo processo de maceração em solução hidroalcoólica 70% por sete dias, à temperatura ambiente. O filtrado resultante da filtração a vácuo foi evaporado sob vácuo a temperatura de 50ºC até a remoção total do etanol, sendo obtido desta forma o extrato bruto hidroalcoólico a frio (EBHF). Os resultados das secções histológicas permitiram a caracterização anatômico-foliar baseada na qualidade das secções manuais obtidas. Os testes em relação às classes de metabólitos secundários revelaram a presença de antocianinas e taninos e ausência de flavonóides e saponinas. Esses resultados estimulam a realização de estudos mais aprofundados para identificar e quantificar os compostos dessas e de outras classes de metabólitos.

In natural medicine, little is known about the healing importance of most of the species belonging to the family Bromeliaceae. Such a fact points tothe need to perform investigations toprovethe possible phytotherapic potentials in species of the family. Guzmania lingulata is characterized as one of the most popular well-known and commercialized species of bromeliads, being widely spread in Brazil. This work aimed at the anatomic characterization and identification of classes of the secondary metabolites present in G. lingulata, by anatomic, histochemical and chemical analysis of the leaves. The leaves were fixed in FAA 50 and, afterwards, transferred to 50% ethanol. For the anatomic study, transverse sections of the middleproportion of the leafblades were obtained by hand and exposed to the process of double coloration with safranin and astra blue, being, right after that, mounted on glycerin jelly. For the sorting of the classes of secondary metabolites, 10 fresh leaves were collected and then put through the maceration process in 70% hydroalcoholic solution for 7 days, at room temperature. The resulting filtered solution from the vacuum filtration was evaporated under vacuum at 40ºC until the complete removal of the ethanol, obtaining, this way, the cold crude hydroalcoholic extract (EBHF). The results ofhistological sectionsallowed the anatomical characterization based onthe quality ofthe manual sectionsobtained. The testsin relationto the classes of secondary metabolitesrevealed the presence ofanthocyanins andtannins andabsence of flavonoids and saponins. These results encourage the completionoffurther studies to identify and quantify the compounds of these and other classes of metabolites.

Histological alterations found in the ureter during organ preservation and early phases of renal transplantation.

INTRODUCTION: There are no studies on the phenomena that occur on the ureter during organ preservation and immediately after transplantation. MATERIAL AND METHODS: We studied ureteral fragments obtained during organ harvesting in the cadaver (n = 9), after cold preservation period (n = 18), and immediately after kidney graft reperfusion (n = 126). In addition to the histological analysis, we evaluated the risk factors for the development of lesions and their relation to the evolution of the transplant. RESULTS: Alterations were detected in 120 of the 126 fragments studied after graft reperfusion. Global cellular infiltration was considered to be normal, mild, and moderate to severe in 34.9%, 41.3%, and 23.8%, respectively, consisting mainly of CD8(+) T lymphocytes. Urothelial exfoliation and cell vacuolization were detected in 42% and 52.4% of the cases, respectively. There was an inverse relationship between donor ventilation time and the intensity of the cellular infiltration. Seven and three of the nine fragments obtained during organ harvesting showed mild cellular infiltration of the chorion and urothelium, respectively. Cold storage promoted minor histological changes. After reperfusion, there was increased urothelial infiltration in 11 of the 18 cases. There was no relation between the lesions encountered and human leukocyte antigen compatibilities, renal rejections episodes, or the evolution of the graft itself. CONCLUSIONS: Consequences of brain death mechanical ventilation were detected at the ureteral level, with abnormal lymphocytic infiltration in most cases. Cold storage did not produce any major histological changes. The lesions detected after graft reperfusion do not seem to involve immunological phenomena.

Ureteric lesions detected in normally functioning kidney allografts: risk factors and clinical implications.

INTRODUCTION: We characterized the alterations in ureteral biopsies from normally functioning kidney allografts to study risk factors. MATERIALS AND METHODS: We studied 55 ureteral fragments from kidney grafts obtained during cystoscopy for routine double-J stent extraction. We evaluated the type and severity of the lesions, the risk factors for their occurrence, and their relation to the evolution of the transplant, including the occurrence of renal rejection episodes or ureteral complications. RESULTS: Borderline or rejection lesions were detected in 21 of the 55 fragments. Rejection lesions were more common among biopsies performed in the first 80 days (54.6%) than during the 120 days afterward (15.4%, P = .043). Similarly, urothelium reactivity was detected in 71.4% and 30.7% of the biopsies performed up to and after 4 months, respectively (P = .008). Urothelial atrophy was detected in 20% of the fragments, the age of the donors being higher in these cases (P = .026). There was a trend to the association of borderline or rejection lesions in the ureteral biopsies and a history of an acute renal rejection episode (P = .053). There were no detectable relations between those findings and the evolution of the transplant. CONCLUSIONS: Thirty-eight percent of the biopsed ureters showed rejection or borderline lesions, these lesions were more common among biopsies done in the first months after transplantation. These findings are similar to the ones found in routine renal biopsies. We did not find any significant relation between the nature of the lesions encountered on the ureteral biopsies and the evolution of the transplant.

Bacterioplankton abundance, biomass and production in a Brazilian coastal lagoon and in two German lakes.

The bacterioplanktonic abundance, biomass, and production within a tropical lagoon (Cabiúnas, Brazil) and two temperate lakes (Stechlin and Dagow, Germany) were compared. Bacterial abundance and production were significantly different among the three water bodies. The lowest bacterial production (0.8 microg C l(-1) d(-1)) was observed in the tropical Cabiúnas Lagoon despite its higher mean temperature and dissolved organic carbon concentration. Highest bacterioplankton abundance (2.6 x 10(9) cells l(-1)) and production (68.5 microg C l(-1) d(-1)) were measured in eutrophic Lake Dagow. In oligotrophic Lake Stechlin, the lowest bacterial biomass (48.05 microg C l(-1)) was observed because of lower bacterial biovolume (0.248 microm(3)) and lower bacterial abundance. Bacterial populations in the temperate lakes show higher activity (production/biomass ratio) than in the tropical lagoon. The meaning of isotopic dilution and leucine incorporation by non-bacterial micro-organisms were evaluated in the oligotrophic temperate system. Leucine uptake by non-bacterial micro-organisms did not have significant influence on bacterial production.

Different methods for extracting bacteria from freshwater sediment and a simple method to measure bacterial production in sediment samples.

The efficiency of different treatments was tested to extract bacterial cells from freshwater sediment samples. The influence of sonication, density gradient centrifugation, fixation by formalin and centrifugation speed on bacterial recovery was investigated. The method developed by Smith and Azam [Mar. Microb. Food Webs 6 (1992) 107] to measure microbial activity on bacterioplankton (3H-leucine incorporation), was also evaluated in sediment samples. After 1 min of sonication bacterial abundance was reduced by about 47% in diluted sediments with tetrasodium pyrophosphate. With the addition of Percoll after sonication, bacterial counts were not significantly different (P<0.05). Fixation by formalin increased bacterial counts using sonication. However, higher bacterial abundance was estimated in non-sonicated samples. Bacterial abundance in samples centrifuged at 7000xg with and without Percoll was not significantly different (P<0.05). Highest bacterial abundance was obtained after centrifugation at low speed (750xg). Bacterial abundance decreased with higher centrifugation speed (750, 1500 and 3000xg), the difference, however, was not significant. Bacterial production ranged from 0.10 microg C cm(-3) d(-1) in autoclaved sediment to 0. 27 microg C cm(-3) d(-1) in untreated sediment. The radioactivity measured in controls of both untreated and autoclaved sediment was high (70 and 91%, respectively), indicating a high level of leucine adsorption in sediment particles. In contrast, radioactivity in control samples previously centrifuged was markedly lower (6%). Despite the high values of radioactivity in the controls, bacterial production in untreated sediment was significantly higher than in centrifuged sediment (P<0.05).

Haemodynamics during liver transplantation in familial amyloidotic polyneuropathy: study of the intraoperative cardiocirculatory data of 50 patients.

BACKGROUND: Circulatory instability frequently complicates liver transplantation for familial amyloidotic polyneuropathy (FAP) and may be a source of surgical morbidity and mortality. OBJECTIVE: To evaluate FAP intraoperative haemodynamic data and their relation to the duration of surgery, and need for anaesthetic drugs. RBC and sympathomimetic amines. SETTING: Clinical study during a four year period. PATIENTS (mean +/- SD): Group I included 50 consecutive FAP ATTR Met 30 recipients of first transplantation. Age was 35.3 +/- 7.1 years, neurological score 34.3 +/- 13 in 100 and time elapsed from first symptom 5.0 +/- 2.7 years. Group II (control), not different concerning age and sex, included 51 patients transplanted during the same period with other pathologies. METHOD: Anaesthetic protocol, monitoring and surgical techniques were similar in both groups. Data of the two groups were compared either by the Student's t-test or Fisher's exact test. RESULTS: Low values of systemic vascular resistance index were observed in both groups, with no differences between them. Systemic arterial pressures were usually lower in group I, because cardiac index and heart rate were also significantly lower, although within normal values. However, in group I, isoflurane (a vasodilator anaesthetic) was used during less time (p < 0.05) and in lower concentrations (p < 0.01) and phenylephrine was necessary in 26% of patients vs 0 patients in group II (p < 0.001). CONCLUSION: FAP patients presented a different intraoperative behaviour when compared to other patients submitted to liver transplantation. From a clinical point of view, the authors stress: 1--As a result of autonomic dysfunction, the administration of anaesthetic drugs to FAP patients always presents the risk of producing significant hypotension; even the use of ketamine does not prevent hypotension; 2--Safety is ensured by beat-to-beat surveillance of arterial pressures and the capacity to act immediately to support circulation; 3--These patients seem to be very sensitive to decreases in the pre-load; 4--Hypotension is also frequent with an adequate pre-load, usually as the result of low SVR; an infusion of a vasoconstrictor drug emerges as the most frequent treatment requested and our experience supports it as an effective one.

Liver transplantation for familial amyloid polyneuropathy.

Familiar Amyloid Polyneuropathy (FAP), an autosomal dominant inherited multisystemic disorder was first observed by Corino de Andrade, a Portuguese neurologist, in 1939. This disease of Portuguese origin was probably spread by fishermen, mainly to Sweden and Japan. It is characterized by a progressive peripheral polyneuropathy and autonomic neuropathy (erectile sexual disfunction, gastrointestinal disfunction, bladder dysfunction and cardio vascular disease) and malnutrition. There are neural and systemic amiloid deposits. Type I FAP, of Portuguese origin, is the most common variety. The amyloid protein is the variant transthyretin (TTR) in which methionine (MET) is a substitute for valine in position 30 (TTR MET 30). It is mainly produced by the liver (90%) and, in small amounts, by the choroidal plexus. Symptoms usually start in the 3rd and 4th decade of life and the patients usually die within 10-15 years. From the therapeutic options--plasmapheresis, immunoadsorption and liver transplantation; the latter seems to be the only one, which stops the production of TTR MET 30 in a permanent way, by means of the liver. The lack of any other effective therapy and the success of the first liver transplantation performed in Sweden arouse great hope. So far, around 300 patients have been transplanted all over the world. A hundred and thirty of them were transplanted in Portugal. A Kaplan Meier survival curve of the Portuguese patients shows a survival rate of 78% at 5 years. However, in spite of the progression of the disease being halted, the irreversibility of some neurological lesions seems to persist. This fact raises the problem of the timing of the transplantation. It seems that the patients should be transplanted as soon as the symptoms start, since mortality and severe morbidity seems to mainly involve those in whom symptomatic disease has lasted longer than six years. As the explanted liver is a morphologic normal liver, a sequential (domino) transplant has been carried out in 16 cases so far done--by one of the authors (ALF) on patients with either hepatocellular carcinoma or liver metastatic disease.

[Kidney transplantation in patients with type I and type II diabetes mellitus]. / Transplantação renal em doentes com diabetes mellitus tipo I e tipo II.

A total of 618 patients with end-stage renal disease received kidney transplants between 1980 and September 1996. Twenty eight of them were diabetics. Better results were achieved for type 1 diabetic patients than for type 2 (mortality: 5.9% vs 27.3%; functioning graft: 88.2% vs 72.7%). The morbility was also higher in those patients (infections: 81.8% vs 29.4%; vascular complications: 45.5% vs 17.6%). Actuarial patient and graft survival were lower for type 2 than for non diabetic patients. For type 1 diabetics the results are similar to those for non diabetics. Better results can probably be achieved by restricting the selection criteria. The decision to transplant or maintain on dialysis should be made on a case by case basis.

Serum potassium concentrations after suxamethonium in patients with familial amyloid polyneuropathy type I.

BACKGROUND: Suxamethonium produces an abnormal increase in serum potassium in some neurological diseases and some authors have suggested that it is safer not to use this drug in patients with familial amyloid polyneuropathy (FAP). However, there are no data previously reported to support this hypothesis. The aim of this study was to evaluate the magnitude of the potassium increase produced by suxamethonium in FAP type I. METHOD: Twenty-one FAP Met 30 patients anaesthetised for liver transplantation were studied. Age was 34.9 +/- 6.9 years (mean +/- SD), time elapsed from first symptom 5.5 +/- 3.2 years and weight was 14 +/- 9% below ideal body weight. Anaesthesia was induced with thiopentone and low-dose fentanyl. Samples for blood gas and 5 min after 1 mg/kg of suxamethonium was given for tracheal intubation. RESULTS: Before induction serum potassium levels were 3.8 +/- 0.4 mmol/L. One minute after suxamethonium, values were 3.8 +/- 0.4 mmol/L and 5 min after 4.3 +/- 0.5 mmol/L. The maximal increase observed was 1.6 mmol/L (from 3.4 mmol/L to 5.0 mmol/L). CONCLUSION: The average increase in plasma potassium concentrations observed in FAP patients after suxamethonium was similar to the increase observed in a normal population by others. Our study can exclude the hypothesis that an anomalous increase in potassium would be a typical and frequent response to suxamethonium in FAP met 30 patients. However, we cannot exclude that a dangerous rise in serum potassium may exist in a certain percentage of FAP patients.

Cerebral toxoplasmosis after renal transplantation. Case report and review.

Infection caused by Toxoplasma gondii is a frequent event in Portugal. When this occurs in immunocompetent individuals, it is rarely a matter of concern; the contrary occurs with immunosuppressed patients or in pregnancy. Transplant patients are treated with immunosuppressive drugs which mainly disturb their mechanisms of cellular immunity, and that opens the way to infections by opportunistic intracellular microorganisms. We recently treated a renal transplant patient who suffered from cerebral toxoplasmosis, and this provided an opportunity for a review of the other 20 patients reported in medical literature to date.