Ontogeny of humoral immunity in northern elephant seal (Mirounga angustirostris) neonates

OBJECTIVE: To assess relations of left ventricular (LV) geometry and function to insulin resistance in obesity-a condition associated with volume overload and abnormal LV relaxation. DESIGN: Cross-sectional relational study. SUBJECTS: 27 healthy overweight-obese subjects (18 women, body mass index (BMI) = 35.0+/-4.0 kg/m2) and 31 age-matched normal-weight controls (21 women, BMI = 22.6+/-2.4 kg/m2). MEASUREMENTS: Subjects were studied by Doppler-echocardiography the same day and hour (08.00 h) as measurements of fasting insulin and blood glucose were made. Insulin resistance was determined by the 'Homeostasis Assessment Model'. RESULTS: Twelve obese subjects with insulin resistance (IR) had higher body size than 15 patients without IR and higher blood pressure than normal-weight controls (all P < 0.01). Relative IR was related to isovolumic relaxation time. This relation was not maintained after controlling for age, blood pressure, weight and height. Isovolumic relaxation time was, however, positively related to diastolic blood pressure, a measure of load, in normal controls (r=0.44) and obese without IR (r=0.62) but not in insulin resistant subjects (r=0.14). CONCLUSION: IR does not independently influence myocardial relaxation in uncomplicated obesity, but modulates the effect of load on active diastole.     

Mutant analysis reveals an alternative pathway for N-linked glycosylation in Drosophila melanogaster

The mas-1 gene of Drosophila melanogaster encodes Golgi mannosidase I (MAS-1), and flies homozygous for small deletions of the gene are viable. They show but few abnormalities and those have a low penetrance [Kerscher, S., Albert, S., Wucherpfennig, D., Heisenberg, M. & Schneuwly, S. (1995) Dev. Biol. 168, 613-626]. Here we sequence the N-linked oligosaccharides associated with a reporter protein, and with membrane proteins prepared from wild type and MAS-1 null organisms. The results show that the null organisms synthesise the same range of oligosaccharides as wild type, albeit with different ratios. There is an accumulation of the Man8GlcNAc2 which is one of the substrates for the MAS-1 enzyme. This supports the suggestion of Kerscher et al. that the lack of severe phenotypic disturbances in the null organisms is due to the presence of an alternative pathway(s), but it also shows that this alternative pathway(s) does not entirely compensate for the normal pathway.     

Risk factors associated with chronic hepatitis C virus infection: limited frequency of an unidentified source of transmission

Acute hypoxaemia is a life-threatening emergency. Diagnosis of the exact aetiology maybe complicated by the presence of pre-existing lung conditions. A case report is presented of a non-intubated patient with a pre-existing lung tumour who developed sudden profound hypoxaemia 3 days after emergency abdominal surgery. Definitive aetiological diagnosis was delayed due to chest X-ray features suggestive of compression and erosion of tumour tissue into the airway. Emergency computerised tomography (CT) imaging however revealed mucous plugging leading to massive atelectasis as the main aetiology.     

Evidence that different mechanisms underlie smooth muscle relaxation to nitric oxide and nitric oxide donors in the rabbit isolated carotid artery

1. The endothelium-dependent relaxants acetylcholine (ACh; 0.03-10 microM) and A23187 (0.03-10 microM), and nitric oxide (NO), applied either as authentic NO (0.01-10 microM) or as the NO donors 3-morpholino-sydnonimine (SIN-1; 0.1-10 microM) and S-nitroso-N-acetylpenicillamine (SNAP; 0.1-10 microM), each evoked concentration-dependent relaxation in phenylephrine stimulated (1-3 microM; mean contraction and depolarization, 45.8+/-5.3 mV and 31.5+/-3.3 mN; n=10) segments of rabbit isolated carotid artery. In each case, relaxation closely correlated with repolarization of the smooth muscle membrane potential and stimulated a maximal reversal of around 95% and 98% of the phenylephrine-induced depolarization and contraction, respectively. 2. In tissues stimulated with 30 mM KCl rather than phenylephrine, smooth muscle hyperpolarization and relaxation to ACh, A23187, authentic NO and the NO donors were dissociated. Whereas the hyperpolarization was reduced by 75-80% to around a total of 10 mV, relaxation was only inhibited by 35% (n=4-7 in each case; P<0.01). The responses which persisted to ACh and A23187 in the presence of 30 mM KCl were abolished by either the NO synthase inhibitor L-NG-nitroarginine methyl ester (L-NAME; 100 microM) or the inhibitor of soluble guanylyl cyclase 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 10 microM; 10 min; n=4 in each case; P<0.01). 3. Exposure to ODQ significantly attenuated both repolarization and relaxation to ACh, A23187 and authentic NO, reducing the maximum changes in both membrane potential and tension to each relaxant to around 60% of control values (n=4 in each case; P<0.01). In contrast, ODQ almost completely inhibited repolarization and relaxation to SIN-1 and SNAP, reducing the maximum responses to around 8% in each case (n=3-5; P<0.01). 4. The potassium channel blockers glibenclamide (10 microM), iberiotoxin (100 nM) and apamin (50 nM), alone or in combination, had no significant effect on relaxation to ACh, A23187, authentic NO, or the NO donors SIN-1 and SNAP (n=4 in each case; P>0.05). Charybdotoxin (ChTX; 50 nM) almost abolished repolarization to ACh (n=4; P<0.01) and inhibited the maximum relaxation to ACh, A23187 and authentic NO each by 30% (n=4-8; P<0.01). Application of ODQ (10 microM; 10 min) abolished the ChTX-insensitive responses to ACh, A23187 and authentic NO (n=4 in each case; P<0.01 5. When the concentration of phenylephrine was reduced (to 0.3-0.5 microM) to ensure the level of smooth muscle contraction was the same as in the absence of potassium channel blocker, ChTX had no effect on the subsequent relaxation to SIN-1 (n=4; P>0.05). However, in the presence of tone induced by 1-3 microM phenylephrine (51.2+/-3.3 mN; n=4), ChTX significantly reduced relaxation to SIN-1 by nearly 50% (maximum relaxation 53.2+/-6.3%, n=4; P<0.01). 6. These data indicate that NO-evoked relaxation of the rabbit isolated carotid artery can be mediated by three distinct mechanisms: (a) a cyclic GMP-dependent, voltage-independent pathway, (b) cyclic GMP-mediated smooth muscle repolarization and (c) cyclic GMP-independent, ChTX-sensitive smooth muscle repolarization. Relaxation and repolarization to both authentic and endothelium-derived NO in this large conduit artery appear to be mediated by parallel cyclic GMP-dependent and -independent pathways. In contrast, relaxation to the NO-donors SIN-1 and SNAP appears to be mediated entirely via cyclic GMP-dependent mechanisms.     

Transient and long-lasting effects of iontophoretically administered norepinephrine on somatosensory cortical neurons in halothane-anesthetized cats

We examined the modulatory effects of iontophoretically administered norepinephrine (NE) on the excitability of 117 neurons in cat somatosensory cortex. NE was released in the vicinity of neurons with receptive fields (31/117) while they were excited by somatic stimuli and near neurons without receptive fields (86/117) while they were excited by glutamate. In 54% of the neurons (n = 63) the effects were inhibitory, decreasing both the spontaneous and the driven activity. Most of these cells were found in the middle layers of cortex. In 36% of the neurons (n = 42), mostly located in either the upper or lower layers, the effects were excitatory, enhancing either or both driven and spontaneous activity, but 52% (n = 22) of this group displayed a transient phase of inhibition. NE usually had a proportionately greater effect on the spontaneous activity than on the evoked activity. Effects of specific NE agonists and antagonists indicated that alpha 2- and beta-receptors mediated the inhibition, but that alpha 1-receptors mediated the excitation. We hypothesize that when NE is released in cat somatosensory cortex, it modulates neuronal responses to afferent activity by generally reducing the excitability of cells in the middle layers and by increasing their signal-to-noise ratios. However, in the upper and lower layers NE will enhance neuronal activity, encouraging exchanges with other cortical areas. In addition, we tested and confirmed the hypothesis that short treatments with NE can modify the excitability of neurons in adult cat somatosensory cortex for long periods of time. Of 69 cells, 59% showed effects of NE lasting for more than 5 min. Response enhancement lasting for the duration of the recording session (5 to 36 min) occurred in 82% (18/22) of silent cells without a receptive field and in 63% (17/27) of spontaneously active cells without a receptive field, but only in 14% (2/14) of cells with a receptive field, suggesting an inverse relationship between cell excitability and this effect of NE. The magnitude of the enhancement followed the same relationship, being greatest for silent cells and least for cells with receptive fields (125, 58, and 49%, respectively). The long-lasting enhancement was blocked by an alpha 1-receptor antagonist in 6 of 9 neurons tested, while the administration of alpha 2- and beta-receptor agonists never produced a long-lasting effect, suggesting that the effect is mediated by alpha 1-adrenoceptors.     

Predicting the future and reconstructing the past: a Bayesian characterization of the utility of subjective fluency

The subjective sense of fluency with which an item can be perceived or remembered is proposed to be a vital cue in making decisions about the future memorability and the nature of our past experience with that stimulus. We first outline a number of cases in which such perceptual or retrieval fluency influences judgments both about our own future performance and our likely past experience, and then present a Bayesian analysis of how judgments of recognition--deciding whether or not a currently viewed item was studied at a particular point in the past--may incorporate information about the perceptual fluency of that item. Using a simple mathematical model, we then provide an interpretation of certain enigmatic phenomena in recognition memory.     

HeLa cells as a model to study the invasiveness and biology of Legionella pneumophila

HeLa cells were established as a model system to study the invasiveness and biology of Legionella pneumophila. In this model, invasion could be distinguished from adherence; virulent strains of L. pneumophila were adherent and invasive, whereas nonvirulent strains were adherent but poorly invasive. Invasion was rapid and did not require de novo bacterial protein synthesis, suggesting that the invasion factor is constitutively expressed by virulent strains. Entry into HeLa cells required actin polymerization and an intact microtubule cytoskeleton and was only moderately inhibited by the presence of 100 mM glucose or galactose. Intracellular replication of virulent L. pneumophila took place in ribosome-studded complex endosomes and led to the formation of free bacteria-laden vesicles presumably released from lysed HeLa cells. These free vesicles (referred to as mature vesicles) were isolated in continuous density gradients of Percoll. The bacteria contained in the isolated mature vesicles had a unique envelope structure and were highly adherent to HeLa cells, characteristics that correlated with a bright red appearance after the Giménez stain (Giménez positive). Plate-grown legionellae and replicating legionellae, harboured in complex endosomes, displayed a typical Gram-negative envelope and stained green after the Giménez stain (Giménez negative). Chronically infected cultures of HeLa cells were also established that may be a useful tool for studying long-term interactions between virulent L. pneumophila and mammalian cells. HeLa cells constitute a valuable model system that offers unique opportunities to study parasite-directed endocytosis, as well as stage specific-parasite interactions.