Negative charge distribution and density on the surface of oxygenated normal and sickle red cells

Negative charges on the external surface of red cells were visualized by colloidal iron hydroxide labelling of 50% of the membrane area after osmotic hemolysis and glutaraldehyde fixation. Counts were made over randomly selected areas on electron micrographs at 350,000 x magnification. Statistical analyses showed that at the 95% level of confidence there was no significant difference between oxygenated normal (AA) and sickle (SS) cells in either the distribution or the density of negative charges.     

Micropropagation, antinociceptive and antioxidant activities of extracts of Verbena litoralis Kunth (Verbenaceae)

This work describes an efficient micropropagation protocol for Verbena litoralis and the study of the antinociceptive and antioxidant activities in extracts of this species. For the establishment in vitro, surface-sterilization procedures and PVPP showed high efficiency in fungal-bacterial contamination and phenol oxidation controls. Nodal segments cultivation in MS medium supplemented with 6-benzyladenine (7.5 μM)/α-naphthaleneacetic acid (NAA; 0.005 μM) induced multiple shoots. Elongated shoots were rooted with IAA (0.2 μM). Acclimatization rates were elevated and the plants showed the typical features of this species. The hexanic fraction (HF) of powdered leaves presented a radical scavenging activity with IC(50) = 169.3 μg mL(-1). HF showed a non-dose dependent analgesic activity in the writhing test; its antinociceptive activity in the hot plate test was restricted to 500 mg kg(-1), which is the highest dose. The results of this study showed the potential of tissue culture on conservation and large scale multiplication and confirmed the traditional folk medicine use of V. litoralis.     

Erythropoietin structure-function relationships: high degree of sequence homology among mammals

To investigate structure-function relationships of erythropoietin (Epo), we have obtained cDNA sequences that encode the mature Epo protein of a variety of mammals. A first set of primers, corresponding to conserved nucleotide sequences between mouse and human DNAs, allowed us to amplify by polymerase chain reaction (PCR) intron 1/exon 2 fragments from genomic DNA of the hamster, cat, lion, dog, horse, sheep, dolphin, and pig. Sequencing of these fragments permitted the design of a second generation of species-specific primers. RNA was prepared from anemic kidneys and reverse-transcribed. Using our battery of species-specific 5' primers, we were able to successfully PCR- amplify Epo cDNA from Rhesus monkey, rat, sheep, dog, cat, and pig. Deduced amino acid sequences of mature Epo proteins from these animals, in combination with known sequences for human, Cynomolgus monkey, and mouse, showed a high degree of homology, which explains the biologic and immunological cross-reactivity that has been observed in a number of species. Human Epo is 91% identical to monkey Epo, 85% to cat and dog Epo, and 80% to 82% to pig, sheep, mouse, and rat Epos. There was full conservation of (1) the disulfide bridge linking the NH2 and COOH termini; (2) N-glycosylation sites; and (3) predicted amphipathic alpha- helices. In contrast, the short disulfide bridge (C29/C33 in humans) is not invariant. Cys33 was replaced by a Pro in rodents. Most of the amino acid replacements were conservative. The C-terminal part of the loop between the C and D helices showed the most variation, with several amino acid substitutions, deletions, and/or insertions. Calculations of maximum parsimony for intron 1/exon 2 sequences as well as coding sequences enabled the construction of cladograms that are in good agreement with known phylogenetic relationships.     

鼓槌石斛中一新的联苄类化合物——鼓槌石斛素

从兰科鼓槌石斛(Dendrobium chrysotoxum)中分得一新的联苄类化合物,经光谱分析(UV,IR,MS,¹HNMR,¹³CNMR和DIFNOE),其结构确定为4-羟基-3,5,3',4'-四甲氧基联苄,命名为鼓槌石斛素。关键词鼓槌石斛;鼓槌石斛素;联苄类化合物。     

A qualitative and quantitative analysis of the entorhinal cortex in schizophrenia

The entorhinal cortex (ERC) has been implicated in schizophrenia by a number of studies. There is anatomical observation of neuronal heterotopias in the rostral ERC, which is consistent with a hypothesis of neurodevelopmental abnormalities in this disease. In view of the significant cytoarchitectonic variation of the ERC throughout its rostro-caudal extent, we performed a detailed subareal analysis of the rostral two-thirds of the entorhinal cortex (ERCr) in 14 postmortem schizophrenic brains and 14 matched controls (mean ages of 48 and 47 respectively). This systematic evaluation included both a qualitative microscopic analysis of morphogenetic anomalies that would be consistent with neurodevelopmental pathology and quantitative measurements of total neuronal number, average neuronal density, laminar volume and laminar depth from the cortical surface in cytoarchitectonically matched subareas of schizophrenic and control brains. Parcellation of the entire ERC on the basis of cytoarchitectonic criteria identified five distinct regions, similar to those described in the macaque, except that in the human brain three of the regions were further divisible into two or three subareas, yielding nine distinct cellular compartments. Five rostral areas, prorhinal (Pr), lateral (28L), intermediate rostral and caudal (281r and 281c), and sulcal (28S), comprise the ERCr. Gross and microscopic examination of these subdivisions throughout the ERCr failed to reveal laminar disorganization in any of the schizophrenic brains. The brains also did not differ significantly with respect to total neuronal number, total volume and neuronal density per laminar and subareal subdivision, or laminar thickness per entorhinal subarea. However, neuronal number and density were reduced by 12-18% in Pr and 28L, suggesting that mild quantitative abnormalities may exist in the ERCr and might possibly be revealed in a larger sample of schizophrenic brains. We have failed to confirm previous reports of laminar disorganization in the ERCr in brains of patients with schizophrenia; to the extent that this region is implicated in schizophrenia, the structural changes are likely to consist of more subtle cellular disturbances.      

Percutaneous ablation of atrial fibrillation: assessment of outcomes at 1-year follow-up.

OBJECTIVE: Current non-pharmacologic treatment strategies for atrial fibrillation (AF) involve discrete ablation of ectopic foci or isolation of the pulmonary vein (PV) ostia, which have been implicated in the genesis of this rhythm disorder. The aim of this study was to determine intermediate and long-term outcomes after percutaneous AF ablation and predictors of successful treatment. POPULATION AND METHODS: We studied 29 consecutive patients (72% male, mean age 52.3 +/- 13 years) who underwent percutaneous ablation of AF refractory to antiarrhythmic drug therapy. AF was idiopathic in 22 patients and the remaining 7 patients were hypertensive. Six patients had permanent AF and the others had recurrent paroxysmal AF (with at least 1 episode a week). Left atrial size was 40.3 +/- 6.1 mm. A total of 35 ablation procedures were performed, 12 using the focal ablation technique and 23 by PV isolation. Six patients underwent a redo procedure. Patients were evaluated at 1-year follow-up (symptoms, medication, ECG and Holter monitoring). Predictors of successful treatment were identified among baseline clinical variables (age, gender, hypertension), LA size, AF sub-type, ablation technique, and number of isolated PV, using multivariable logistic regression. RESULTS: At 1-year follow-up, 20 patients presented sinus rhythm (69%), of whom 7 were taking antiarrhythmic drugs and remained free of AF relapse. Out of the total of 35 procedures, 2 immediate complications occurred: pericardial tamponade in one patient and right phrenic nerve palsy in another. Predictors of long-term success were absence of a prior history of hypertension and isolation of at least three PV (p = 0.01 for both independent predictors). CONCLUSION: Two out of three patients who underwent AF ablation presented sinus rhythm one year after the intervention. Isolation of at least three PV and idiopathic etiology are independent predictors of successful treatment.