Temporal profile of potassium channel dysfunction in cerebrovascular smooth muscle after experimental subarachnoid haemorrhage

The pathogenesis of cerebral vasospasm after subarachnoid haemorrhage (SAH) involves sustained contraction of arterial smooth muscle cells that is maximal 6–8 days after SAH. We reported that function of voltage-gated K⁺ (K_V) channels was significantly decreased during vasospasm 7 days after SAH in dogs. Since arterial constriction is regulated by membrane potential that in turn is determined predominately by K⁺ conductance, the compromised K⁺ channel dysfunction may cause vasospasm. Additional support for this hypothesis would be demonstration that K⁺ channel dysfunction is temporally coincident with vasospasm. To test this hypothesis, SAH was created using the double haemorrhage model in dogs and smooth muscle cells from the basilar artery, which develops vasospasm, were isolated 4 days (early vasospasm), 7 days (during vasospasm) and 21 days (after vasospasm) after SAH and studied using patch-clamp electrophysiology. We investigated the two main K⁺ channels (K_V and large-conductance voltage/Ca²⁺-activated (K_Ca) channels). Electrophysiologic function of K_Ca channels was preserved at all times after SAH. In contrast, function of K_V channels was significantly decreased at all times after SAH. The decrease in cell size and degree of K_V channel dysfunction was maximal 7 days after SAH. The results suggest that K_V channel dysfunction either only partially contributes to vasospasm after SAH or that compensatory mechanisms develop that lead to resolution of vasospasm before K_V channels recover their function.     

Dendritic cells infected with Mycobacterium bovis bacillus Calmette Guerin activate CD8(+) T cells with specificity for a novel mycobacterial epitope

Although CD4(+) T cells are essential for protective immunity against Mycobacterium tuberculosis infection, recent reports indicate that CD8(+) T cells may also play a critical role in the control of this infection. However, the epitope specificity and the mechanisms of activation of mycobacteria-reactive CD8(+) T cells are poorly characterized. In order to study the CD8(+) T cell responses to the model mycobacterial antigen, MPT64, we used recombinant vaccinia virus expressing MPT64 (VVWR-64) and a panel of MPT64-derived peptides to establish that the peptide MPT64(190-198) contains an H-2D(b)-restricted CD8(+) T cell epitope. A cytotoxic T lymphocyte response to this peptide could be demonstrated in M. bovis bacillus Calmette Guerin (BCG)-infected mice following repeated in vitro stimulation. When bone marrow-derived dendritic cells (DC) were infected with BCG, the expression of MHC class I molecules by DC was up-regulated in parallel with MHC class II and B7-2, whereas CD1d expression level was not modified. Moreover, BCG-infected DC activated MPT64(190-198)-specific CD8(+) T cells to secrete IFN-gamma, although with a lower efficacy than VVWR-64-infected DC. The production of IFN-gamma by MPT64(190-198)-specific CD8(+) T cells was inhibited by antibodies to MHC class I, but not to CD1d. These data suggest that mycobacteria-specific CD8(+) T cells are primed during infection. Therefore, anti-mycobacterial vaccine strategies targeting the activation of specific CD8(+) T cells by DC may have improved protective efficacy.     

Influence of rifampin therapy on serum bactericidal activity in the presence of cloxacillin and vancomycin

In this study the effect of rifampin on serum inhibitory and serum bactericidal titres was examined. Sera were prepared from pooled human serum to contain vancomycin (10 mg/L), cloxacillin (5 mg/L) or rifampin (1 mg/L), and the combinations cloxacillin/rifampin and vancomycin/rifampin. These five sera were tested by a microtitre method for serum inhibitory power and serum bactericidal titre against 11 strains of Staphylococcus aureus. A 48 h incubation period was required to detect full colony growth for subculture plates. It was found with all strains that the effect of the addition of rifampin to the other two antibiotics was to increase the serum inhibitory power, lower the serum bactericidal titre, increase the inhibitory/cidal ratio, and slow colony growth on subculture. In the clinical part of the study it was shown that only three of 38 sera (8%) from patients receiving betalactam or vanomycin but not rifampin gave an inhibitory/cidal ratio greater than 8, but that nine of 10 sera (90%) from patients receiving rifampin in addition to betalactam or vancomycin gave a ratio greater than 8 (P<0.001). The study verified that the effect of rifampin in serum was to increase inhibitory power and decrease bactericidal titre. The clinical significance of these results is not known and it is suggested that a high ratio of inhibitory to bactericidal titre in the presence of rifampin is to be expected, and that a low bactericidal titre under these circumstances is not necessarily an indication to modify therapy.     

Clinical and genetic features of variegate porphyria in a Chinese patient

Acute porphyria is rare in orientals. We describe a Chinese woman with recurrent generalised tonic-clonic seizures and abdominal pain. Genomic DNA studies identified a heterozygous base substitution from guanine to adenine at nucleotide position 503, resulting in substitution of arginine by histidine at position 168 of the protein (R168H). This genetic abnormality is similar to the mutation reported in Caucasians with variegate porphyria. To the best of our knowledge, this is the first report in the English literature a Chinese patient with variegate porphyria with an identifiable mutation. A brief review of porphyria is presented.     

Work-place alcohol and other drug testing: a review of the scientific evidence

In this paper, scientific evidence for alcohol and other drug testing programs is compared to determine whether each approach is justifiable for improving work-place safety. Three types of studies are reviewed: laboratory, epidemiological and evaluation studies. Laboratory studies show that alcohol use decreases psycho-motor performance; however, for other drugs, some drugs deteriorate performance while others have little effect. Epidemiological studies in the work-place have not provided conclusive evidence that a strong causal link exists between either alcohol or other drug use and work-place injuries/accidents. Evaluation studies have not shown that either drug or alcohol testing significantly reduces work injuries/accidents. Other types of scientific evidence provide some justification of alcohol testing, but not for other drug testing. Alcohol use is much more common than other drug use in industrialized countries, suggesting that alcohol may be more related to industrial accidents than other drugs. As well, epidemiological studies suggest that alcohol is a major factor for traffic collisions, but research is inconclusive for other drugs. Finally, alcohol testing is more justifiable than drug testing because the results of alcohol tests closely correlate with psycho-motor performance while drug tests do not.     

Characterization of the P2 receptors in rabbit pulmonary artery

1. We have identified the P₂ receptors mediating vasomotor responses in the rabbit pulmonary artery.
2. Neither ATP nor UTP contracted intact or endothelium-denuded rings. However, both relaxed intact rings of rabbit pulmonary artery that had been preconstricted with phenylephrine (pD₂ 5.2 and 5.6, respectively).
3. The vasodilator effect of UTP was endothelium-dependent and abolished by the nitric oxide synthase inhibitor N^G-nitro-L-arginine (L-NOARG).
4. The vasodilator effect of ATP was only partially inhibited by removal of endothelium or addition of L-NOARG, suggesting an additional direct effect on vascular smooth muscle.
5. The endothelium-dependent vasodilator responses to UTP and ATP were competitively antagonized by suramin.
6. Preconstricted, endothelium-denuded rings were also relaxed by 2-methylthio ATP (pD₂ 6.6), a P_2Y receptor agonist.
7. Ca²⁺-mobilizing P_2U receptors were identified on smooth muscle cells on the basis of single cell responses to ATP (pD₂ 7.8) and UTP (pD₂ 7.9; 6.7 in the presence of 100 μM suramin).
8. There was no evidence of a Ca²⁺-mobilizing P_2Y receptor in these cultured cells.
9. The data suggest the presence of (i) a suramin-sensitive P_2U receptor on endothelial cells that induces vasorelaxation through NO release, (ii) a suramin-sensitive P_2U receptor on cultured smooth muscle cells that mobilizes Ca²⁺ but is not coupled to vasomotor responses and (iii) a putative P_2Y receptor on vascular smooth muscle cells that induces relaxation via a Ca²⁺-independent signal transduction pathway.

     

A comparison of skinfold thickness, body mass index, bioelectrical impedance analysis and dual-energy X-ray absorptiometry in assessing body composition in obese subjects before and after weight loss

The assessment of body composition in obese subjects by anthropometric means (skinfolds), presents many difficulties. This study compares the estimates provided of fat free mass in 21 obese subjects (mean body mass index 36.6 +/- 1.2 kg/m(2)) using body mass index and skinfold thickness, with those obtained using the more recently developed techniques of bioelectrical impedance analysis and dual-energy X-ray absorptiometry. Despite highly significant correlations between some of the methods (r(2) = 0.94 for dual-energy X-ray absorptiometry versus bioelectrical impedance analysis), there was a considerable lack of agreement in the measurements, particularly when skinfold thickness was compared with dual-energy X-ray absorptiometry (limits of agreement -21.9 to -1.5 kg for fat free mass estimated from dual-energy X-ray absorptiometry and skinfold thickness) and even for dual-energy X-ray absorptiometry and bioelectrical impedance analysis (limits of agreement -10.7 to 0.4 kg). After weight loss the intermethod differences were reduced.     

Effect of probenecid on the formation and elimination kinetics of the sulphate and glucuronide conjugates of diflunisal

The effect of probenecid on the pharmacokinetics of diflunisal and its glucuronide and sulphate conjugates was studied in 8 healthy volunteers. Diflunisal 250 mg b. d. was administered p. o. for 15 days and its steady state pharmacokinetics was evaluated on Day 16 after the last dose (control phase). Probenecid 500 mg b. d. was co-administered throughout the entire study period in the treatment phase of the study.The steady state plasma concentration of diflunisal was significantly higher during the probenecid treatment phase as compared to the control phase (104.0 vs. 63.1 g·ml^–1). This was the result of a significant decrease in the plasma clearance of diflunisal from 5.8 (control) to 3.4 ml·min^–1 (probenecid co-administration). The metabolite formation clearances of both glucuronides were significantly decreased by probenecid, -45 % and -54 % for the phenolic and acyl glucuronide, respectively. The metabolite formation clearance of the sulphate conjugate was not affected by probenecid co-administration.Steady state plasma concentrations of the sulphate and glucuronide conjugates of diflunisal were 2.5- to 3.1-fold higher during probenecid co-administration, due to a significant reduction in the renal clearance of the three diflunisal conjugates. Probenecid also reduced the plasma protein binding of diflunisal, but only to a minor extent; the unbound plasma fraction of diflunisal at steady state averaged between 5 and 30 % higher during probenecid co-administration.