STUDIES ON THE SITE OF MEDIATED TRANSPORT OF CHLORIDE FROM BLOOD INTO CEREBROSPINAL FLUID: EFFECTS OF ACETAZOLAMIDE

Abstract— In monkeys we measured the steady-state concentrations of Cl⁻ in endogenous CSF, in artificial CSF (which had equilibrated with the underlying exposed surface of the cerebral cortex but was not in diffusion equilibrium with endogenous CSF), and in arterial plasma. The ratio of the distribution of Cl⁻ in artificial CSF to that in plasma was consistent with a passive Donnan distribution, whereas that ratio describing Cl⁻ levels in endogenous CSF in comparison to those in plasma clearly exceeded theratio required for a passive, Donnan⁻type of distribution for Cl⁻. The kinetic analysis of the efflux of Cl⁻ from blood into endogenous CSF and into artificial CSF (perfused over the exposed surface of the cerebral cortex) indicated that the rate of efflux of Cl⁻ into endogenous CSF which was continuous with ventricular fluid was inhibited by acetazolamide [in confirmation of a similar finding described previously by M aren and B roder (1970)], whereas the rate of efflux of chloride from blood into the artificial CSF perfusate was uninfluenced by pretreatment of animals with acetazolamide. We have discussed the site of mediated (active) transport of chloride from blood into CSF in light of these findings.     

Fluid compartmentation and electrolytes of cat cerebral cortex in vitro. I. Swelling and solute distribution in mature cerebral cortex

The distribution of indicator solutes (inulin, sucrose, raffinose, thiocyanate, chloride, and isethionate) and the swelling in slices of adult cat cerebral cortex incubated in vitro have been investigated for a variety of incubation media and conditions. Where appropriate, these data have been compared with analogous data obtained on cat cerebral cortex in vivo (Bourke et al., 1965) and with data previously reported for slices of rat cerebral cortex (Pappius and Elliott , 1956; Pappius et al., 1962) and guinea pig cerebral cortex (Varon and Mc Ilwain , 1961; Keesey et al., 1965) incubated in vitro under comparable conditions. In addition to substantial agreement of present data with previously reported in vitro data, a number of new findings have been added by the present study: (a) a component of slice swelling which is K⁺-dependent; (b) the association of slice swelling with presence of a diffusible, external anion (Cl^?) and its prevention by replacement with a relatively non-diffusible anion (isethionate^?); (c) variation of the size of slice chloride spaces as a direct function of slice swelling; (d) dependence of the size of slice fluid spaces accessible to inulin and sucrose upon time, during incubation, of solute addition and upon K⁺ concentration of the incubation medium; (e) indications of the dynamic and presumably metabolically-dependent nature of indicator solute distribution; and (f) the mobility of a portion of the fluid of swelling associated with changing the K⁺ concentration but not the tonicity of the medium during incubation. At least five operationally-defined fluid compartments may be inferred from the present data : (1) interstitial or extracellular space(s) readily accessible to all solutes and of a size which can be minimally estimated from direct determinations in viuo; (2) additional fluid space(s) accessible to most solutes, including inulin and sucrose, under apparently suboptimal conditions of slice metabolism in vitro; (3) fluid space(s) prone to swell under in vitro conditions and readily accessible in vitro to chloride and thiocyanate but not to inulin or sucrose; (4) fluid space(s) which swell reversibly in the presence of added external K⁺ (or Rb⁺) and are inaccessible to all usual indicator solutes; and (5) after exclusion of the foregoing, the remaining fluid presumably comprising most of the intracellular space(s). The data have been discussed in terms of the morphological complexity of cerebral cortex, in terms of applicability to studies of cortical electrolyte distribution, and in terms of the general problem of delineating cerebral interstitial spaces.     

Sex investment ratios in eusocial Hymenoptera support inclusive fitness theory

Inclusive fitness theory predicts that sex investment ratios in eusocial Hymenoptera are a function of the relatedness asymmetry (relative relatedness to females and males) of the individuals controlling sex allocation. In monogynous ants (with one queen per colony), assuming worker control, the theory therefore predicts female‐biased sex investment ratios, as found in natural populations. Recently, E.O. Wilson and M.A. Nowak criticized this explanation and presented an alternative hypothesis. The Wilson–Nowak sex ratio hypothesis proposes that, in monogynous ants, there is selection for a 1 : 1 numerical sex ratio to avoid males remaining unmated, which, given queens exceed males in size, results in a female‐biased sex investment ratio. The hypothesis also asserts that, contrary to inclusive fitness theory, queens not workers control sex allocation and queen–worker conflict over sex allocation is absent. Here, I argue that the Wilson–Nowak sex ratio hypothesis is flawed because it contradicts Fisher's sex ratio theory, which shows that selection on sex ratio does not maximize the number of mated offspring and that the sex ratio proposed by the hypothesis is not an equilibrium for the queen. In addition, the hypothesis is not supported by empirical evidence, as it fails to explain ‘split’ (bimodal) sex ratios or data showing queen and worker control and ongoing queen–worker conflict. By contrast, these phenomena match predictions of inclusive fitness theory. Hence, the Wilson–Nowak sex ratio hypothesis fails both as an alternative hypothesis for sex investment ratios in eusocial Hymenoptera and as a critique of inclusive fitness theory.     

Assessing the microbial oxidative stress mechanism of ozone treatment through the responses of Escherichia coli mutants

Aims:  To investigate the effect of the oxidative stress of ozone on the microbial inactivation, cell membrane integrity and permeability and morphology changes of Escherichia coli. Methods and Results:  Escherichia coli BW 25113 and its isogenic mutants in soxR, soxS, oxyR, rpoS and dnaK genes were treated with ozone at a concentration of 6 μg ml^?1 for a period up to 240 s. A significant effect of ozone exposure on microbial inactivation was observed. After ozonation, minor effects on the cell membrane integrity and permeability were observed, while scanning electron microscopy analysis showed slightly altered cell surface structure. Conclusions:  The results of this study suggest that cell lysis was not the major mechanism of microbial inactivation. The deletion of oxidative stress–related genes resulted in increased susceptibility of E. coli cells to ozone treatment, implying that they play an important role for protection against the radicals produced by ozone. However, DnaK that has previously been shown to protect against oxidative stress did not protect against ozone treatment in this study. Furthermore, RpoS was important for the survival against ozone. Significance and Impact of the Study:  This study provides important information about the role of oxidative stress in the responses of E. coli during ozonation.     

Ribavirin enhances IFN-α signalling and MxA expression: a novel immune modulation mechanism during treatment of HCV

The nucleoside analogue Ribavirin significantly increases patient response to IFN-α treatment of HCV, by directly inhibiting viral replication. Recent studies indicate that Ribavirin also regulates immunity and we propose that Ribavirin enhances specific interferon sensitive gene (ISG) expression by amplifying the IFN-α-JAK/STAT pathway. We found that IFN-α-induced STAT1 and STAT3 phosphorylation was increased in hepatocytes co-treated with Ribavirin and IFN-α, compared to IFN-α alone. Ribavirin specifically enhanced IFN-α induced mRNA and protein of the anti-viral mediator MxA, which co-localised with HCV core protein. These novel findings indicate for the first time that Ribavirin, in addition to its viral incorporation, also enhances IFN-α-JAK/STAT signalling, leading to a novel MxA-mediated immuno-modulatory mechanism that may enhance IFN-α anti-viral activity against HCV.     

Volatile biomarkers of Pseudomonas aeruginosa in cystic fibrosis and noncystic fibrosis bronchiectasis

Aims: The purpose of this study was to determine whether volatile organic compounds specific to Pseudomonas aeruginosa could be detected in clinical sputum specimens. Methods and Results: Patients were recruited from specialist bronchiectasis and cystic fibrosis clinics. The gold standard for diagnosing Ps. aeruginosa infection was a positive sputum culture. About 72 sputum headspace samples taken from patients at risk of or known to have prior Ps. aeruginosa infection were analysed by solid phase micro‐extraction mass spectrometry. 2‐nonanone was a marker in Ps. aeruginosa in sputum headspace gas with sensitivity of 72% and specificity of 88%. A combination of volatile compounds, a sputum library of 17 compounds with 2‐nonanone, increased sensitivity in the detection of Ps. aeruginosa to 91% with specificity of 88%. Conclusions: In contrast to the 48‐hour turnaround for classical microbiological culture, these results were available within 1–2 h. These data demonstrate the potential for rapid and accurate diagnosis of Ps. aeruginosa infection from sputum samples. Significance and impact of the study: 2‐Nonanone is a compound requiring further study in the exhaled breath as it may improve diagnostic of Ps. aeruginosa infection when combined with other reported volatile markers.