Isolation and characterization of the outer membrane of Borrelia hermsii

The outer membrane of Borrelia hermsii has been shown by freeze-fracture analysis to contain a low density of membrane-spanning outer membrane proteins which have not yet been isolated or identified. In this study, we report the purification of outer membrane vesicles (OMV) from B. hermsii HS-1 and the subsequent identification of their constituent outer membrane proteins. The B. hermsii outer membranes were released by vigorous vortexing of whole organisms in low-pH, hypotonic citrate buffer and isolated by isopycnic sucrose gradient centrifugation. The isolated OMV exhibited porin activities ranging from 0.2 to 7.2 nS, consistent with their outer membrane origin. Purified OMV were shown to be relatively free of inner membrane contamination by the absence of measurable beta-NADH oxidase activity and the absence of protoplasmic cylinder-associated proteins observed by Coomassie blue staining. Approximately 60 protein spots (some of which are putative isoelectric isomers) with 25 distinct molecular weights were identified as constituents of the OMV enrichment. The majority of these proteins were also shown to be antigenic with sera from B. hermsii-infected mice. Seven of these antigenic proteins were labeled with [3H]palmitate, including the surface-exposed glycerophosphodiester phosphodiesterase, the variable major proteins 7 and 33, and proteins of 15, 17, 38, 42, and 67 kDa, indicating that they are lipoprotein constituents of the outer membrane. In addition, immunoblot analysis of the OMV probed with antiserum to the Borrelia garinii surface-exposed p66/Oms66 porin protein demonstrated the presence of a p66 (Oms66) outer membrane homolog. Treatment of intact B. hermsii with proteinase K resulted in the partial proteolysis of the Oms66/p66 homolog, indicating that it is surface exposed. This identification and characterization of the OMV proteins should aid in further studies of pathogenesis and immunity of tick-borne relapsing fever.     

The effects of starch and protein degradation rates, hay sources, and feeding frequency on rumen microbial fermentation in a continuous culture system

Six continuous culture fermenters were used in three experiments to study the effects of dietary starch and protein degradability combination, hay sources, and feeding frequency on fermentation by rumen microorganisms. Experiments 1 and 2 used a 3 x 2 factorial design in which six diets were formulated to contain low (LS), medium (MS), or high starch degradability (HS) in combination with low (LP) or high protein degradability (HP). The dietary combinations were (1) LS + HP, (2) LS + LP, (3) MS + HP, (4) MS + LP, (5) HS + HP, and (6) HS + LP. In experiment 1, pangola was used as the hay source, and in experiment 2, alfalfa hay was used. In experiment 3, two starch degradabilities (LS, MS) and two hay sources (alfalfa, A; pangola, P) were combined with two feeding frequencies (2 X/d, 12 X/d). The dietary combinations were (1) LS + A + 12 X/d, (2) MS + A + 12 X/d, (3) LS + A + 2 X/d, (4) MS + A + 2 X/d, (5) LS + P + 12 X/d, and (6) MS + P + 12 X/d. A CRD design was used for experiment 3. Two rumen-cannulated Holstein cows fed alfalfa hay ad lib were used as donors of rumen fluid for all experiments. Each period was 6 d in length, with 5 d for adaptation and 1 d for sampling. In experiments 1 and 2, the effects of starch degradability on the composition of rumen microorganisms were significant. The MS or HS with HP had the highest total bacterial and protozoal density (P < 0.05). As for VFA, pH and nitrogen products, the effects of starch and protein degradability were not significant. There was no interaction between starch and protein degradability on most of microbial composition (ammonia-N, microbial nitrogen). In experiment 3, 12 X/d feeding frequency (MS + A + 12 X/d vs. MS + A + 12 X/d) resulted in higher pH, which tended to increase bacterial and protozoal density and starch and protein digestibilities. Different hay sources altered the starch and protein synchronization effect on the ammonia-N concentration (mg/dl) and non-ammonia N content (% DM) in the continuous culture system.     

Pregnancies of women with epilepsy: a population-based study in Iceland

PURPOSE: Women with epilepsy who become pregnant are commonly considered to be at high risk for complications during pregnancy or delivery. The offspring are also considered to have increased risk of perinatal mortality, congenital malformations, and maturational delay. Because few of these studies are population based, potential bias exists because of selection. METHODS: We performed a historical population-based cohort study in Iceland to determine the prevalence of epilepsy among pregnant women, to identify pregnancy and delivery complications in women with epilepsy, and to determine the outcome of their pregnancies as compared with that in the general population of Iceland. We identified all women with active epilepsy who gave birth during a 19-year period in Iceland. RESULTS: In this population, 3.3 in 1,000 pregnancies involve mothers with active epilepsy. The frequency of adverse events (AE) during pregnancy in the women with epilepsy is similar to that observed among all live births in the population, but cesarean section was performed twice as frequently as in the general population. Perinatal mortality rate and mean birth weight are not significantly different in the offspring of women with epilepsy as compared with rest of the population. The risk of major congenital malformations (MGM) is increased 2.7-fold over that expected when a mother is treated with antiepileptic drugs (AEDs) during a pregnancy. CONCLUSIONS: Our study indicates that the rate of complications of pregnancy in mothers with active epilepsy is low and similar to that of the general population with epilepsy. Use of AEDs by the mother during pregnancy significantly increases the risk of MGM in the offspring.     

Fetal cerebral metabolism: the influence of asphyxia and other factors

Cerebral oxidative metabolism has been described in fetal sheep at two stages of development and is known to remain relatively constant over a wide range of oxygen levels in arterial blood. This constancy of oxygen consumption is caused by an increase in cerebral blood flow that matches the reduction in oxygen content and oxygen extraction. Although a number of factors are involved in the hypoxia-associated vasodilation (for example, oxygen, carbon dioxide, adenosine, prostaglandins, arginine vasopressin), its regulation is incompletely understood. During severe asphyxia, however, there is a limit to the vasodilator function, and both cerebral blood flow and oxygen consumption fall. The fetus can tolerate a certain degree of reduced oxygen uptake (possibly to 50% of control level) by various conservation techniques, but severe reductions are associated with neuronal damage. The primary substrate for the fetal brain under normal conditions is glucose, but the fetus can readily use anaerobic glycolysis and produce lactate under conditions of oxygen limitation. Lactate efflux from the brain is relatively slow, so prolonged and severe asphyxia may result in a high tissue level of lactate, which has been implicated in neuronal damage.     

Mechanical circulatory support in children

Nine children (aged 1.2-15 years) have been treated with mechanical circulatory support devices at our institution. Indications for treatment were acute cardiac allograft rejection (n = 4), postcardiotomy cardiogenic shock (n = 4), and bridge to cardiac transplantation (n = 1). Eight patients required left ventricular support, and one required biventricular support. A BioMedicus centrifugal pump was used in eight patients, and a Hemopump intra-aortic axial flow device was used in one patient. In two patients, an intra-aortic balloon pump was in place at the time that circulatory support was instituted. Mechanical support time ranged from 2 to 139 h, and the average flow index was 2.31 l/min per m2. Three patients required hemodialysis during support, and one patient required re-exploration because of mediastinal hemorrhage. Recovery of native ventricular function was assessed by transthoracic or transesophageal echocardiography, and weaning from the device was achieved by gradually decreasing pump flow in increments of 0.1 to 0.5 l/min. Seven patients were successfully weaned from support. Two hospital deaths occurred after circulatory support had been discontinued: one patient died of respiratory failure and the other of gram-negative pneumonia and sepsis. The five surviving patients experienced no significant complications, and their hemodynamic indices were normal at the time of discharge. At a mean follow-up of 28.8 months, these patients are leading active unrestricted lives, with no long-term device-related sequelae. Based on this experience, mechanical circulatory support is feasible in children who experience profound circulatory failure from a variety of causes.     

Intravascular histiocytosis

The majority of cases of intravascular lymphomatosis are B-cell lymphomas with only the occasional case being of T-cell type. We report a case of intravascular lymphomatosis in which the proliferating cells were of histiocytic type; the tumour has recurred following treatment.     

Measurements and a model of the outer hair cell hydraulic conductivity

The dimensions of the apical surfaces of hair cells were measured in guinea pigs, aged from 3 weeks before term to 25 weeks after birth. In the basal two-thirds of the cochlea, the apical surfaces of the outer hair cells and their supporting cells changed with age, shrinking in a direction radial across the cochlear duct. There was an associated widening of the angle of the 'V' of the rows of stereocilia. Further apically, between 12 and 16 mm from the base of the cochlea, the outer hair cells and their supporting cells underwent the opposite change, becoming wider in a radial direction with age. The changes were seen before birth and continued for more than 3 weeks after birth. The results suggest that the guinea pig cochlea continues certain developmental processes for a considerable time after birth.     

A MODEL TO PREDICT THE FATIGUE LIFE OF FIBRE-REINFORCED TITANIUM MATRIX COMPOSITES UNDER CONSTANT AMPLITUDE LOADING

A model based on micro-mechanical concepts has been developed for predicting fatigue crack growth in titanium alloy matrix composites. In terms of the model, the crack system is composed of three zones: the crack, the plastic zone and the fibre. Crack tip plasticity is constrained by the fibres and remains so until certain conditions are met. The condition for crack propagation is that fibre constraint is overcome when the stress at the location of the fibre ahead of the crack tip attains a critical level required for debonding. Crack tip plasticity then increases and the crack is able to propagate round the fibre. The debonding stress is calculated using the shear lag model from values of interfacial shear strength and embedded fibre length published in the literature. If the fibres in the crack wake remain unbroken, friction stresses on the crack flanks are generated, as a result of the matrix sliding along the fibres. The friction stresses (known as the bridging effect) shield the crack tip from the remote stress, reducing the crack growth relative to that of the matrix alone. The bridging stress is calculated by adding together the friction stresses, at each fibre row bridging the crack, which are assumed to be a function of crack opening displacement and sliding distance at each row. The friction stresses at each fibre row will increase as the crack propagates further until a critical level for fibre failure is reached. Fibre failure is modelled through Weibull statistics and published experimental results. Fibre failure will reduce the bridging effect and increase the crack propagation rate. Calculated fatigue lives and crack propagation rates are compared with experimental results for three different materials (32% SCS6/Ti-15-3, 32% and 38% SCS6/Ti-6-4) subjected to mode I fatigue loading. The good agreement shown by these comparisons demonstrates the applicability of the model to predict the fatigue damage in Ti-based MMCs.     

The effect of bonded and nonbonded posts on the fracture resistance of dentin]

This in vitro study determined the effect of bonded and non-bonded cast, serrated, tin plated, parallel gold posts on the fracture resistance of surrounding dentine walls. Specimens were divided into two groups. One group's posts were luted with zinc phosphate cement and the other group's posts were tin-plated and adhesively bonded with an adhesive resin cement. Three different dentine wall thicknesses, namely 0.5 mm, 0.75 mm and 1.0 mm were prepared, subdividing the specimens of each main group into three subgroups. Specimens were subjected to a 50 degrees oblique force. The force required to fracture the dentinal walls was registered. There was no statistically significant differences in the fracture resistance of dentine with the two different cements or of the different dentine wall thicknesses.