Closed recirculatory spinal subarachnoid perfusion for determining CSF dynamics

1982 ◽  
Vol 56 (3) ◽  
pp. 368-372 ◽  
Author(s):  
Alan A. Artru ◽  
Michael Nugent ◽  
John D. Michenfelder

✓ A new method for determining rates of cerebrospinal fluid (CSF) production under nonsteady-state conditions, namely, closed recirculatory spinal subarachnoid perfusion, was used to determine the effect of enflurane on the rate of CSF production in dogs. Considerable variability in results was observed such that there was no statistical difference in rates of production among animals that received enflurane 2.2%, enflurane 2.2% and nitrous oxide 60% to 70%, enflurane 3.2% and nitrous oxide 60% to 70%, or nitrous oxide 60% to 70% (controls). Possible sources of variability were sought in additional studies using a modification of the new method, and in an in vitro model. The results were compared to those obtained using an established method for determining rates of CSF production, namely, open ventriculocisternal perfusion. It was concluded that the sources of variability in the closed recirculatory method relate in part to adherence of the fluorescein-conjugated albumin tracer to glass and other surfaces, and to uneven flow and distribution of the tracer in the recirculatory system. When the open ventriculocisternal perfusion method was used, consistent results were obtained, demonstrating that CSF production rate increased significantly in animals that received enflurane. The authors conclude that the new closed recirculatory method is less reliable than the classical open perfusion method.

1979 ◽  
Vol 50 (4) ◽  
pp. 477-482 ◽  
Author(s):  
A. Richard Vela ◽  
Michael E. Carey ◽  
Bruce M. Thompson

✓ Considerable difference of opinion has arisen as to whether intravenously administered steroids affect cerebrospinal fluid (CSF) production in the acute laboratory animal undergoing ventriculocisternal perfusion. Our experiments with ventriculocisternal perfusion in dogs indicate that, when given intravenously, neither dexamethasone, methylprednisolone, hydrocortisone, nor aldosterone result in a significant, acute effect upon CSF production. Similarly, CSF absorption and outflow resistance mechanisms are not acutely affected by intravenous methylprednisolone, hydrocortisone, and aldosterone. Dexamethasone also probably does not produce an immediate effect upon CSF absorption.


Microbiology ◽  
2020 ◽  
Vol 166 (10) ◽  
pp. 909-917 ◽  
Author(s):  
Sophie P. Bennett ◽  
Maria J. Torres ◽  
Manuel J. Soriano-Laguna ◽  
David J. Richardson ◽  
Andrew J. Gates ◽  
...  

Nitrous oxide (N2O) is a potent greenhouse gas that is produced naturally as an intermediate during the process of denitrification carried out by some soil bacteria. It is consumed by nitrous oxide reductase (N2OR), the terminal enzyme of the denitrification pathway, which catalyses a reduction reaction to generate dinitrogen. N2OR contains two important copper cofactors (CuA and CuZ centres) that are essential for activity, and in copper-limited environments, N2OR fails to function, contributing to rising levels of atmospheric N2O and a major environmental challenge. Here we report studies of nosX, one of eight genes in the nos cluster of the soil dwelling α-proteobaterium Paraccocus denitrificans. A P. denitrificans ΔnosX deletion mutant failed to reduce N2O under both copper-sufficient and copper-limited conditions, demonstrating that NosX plays an essential role in N2OR activity. N2OR isolated from nosX-deficient cells was found to be unaffected in terms of the assembly of its copper cofactors, and to be active in in vitro assays, indicating that NosX is not required for the maturation of the enzyme; in particular, it plays no part in the assembly of either of the CuA and CuZ centres. Furthermore, quantitative Reverse Transcription PCR (qRT-PCR) studies showed that NosX does not significantly affect the expression of the N2OR-encoding nosZ gene. NosX is a homologue of the FAD-binding protein ApbE from Pseudomonas stutzeri , which functions in the flavinylation of another N2OR accessory protein, NosR. Thus, it is likely that NosX is a system-specific maturation factor of NosR, and so is indirectly involved in maintaining the reaction cycle of N2OR and cellular N2O reduction.


1985 ◽  
Vol 63 (5) ◽  
pp. 759-762 ◽  
Author(s):  
Huda Y. Zoghbi ◽  
Sada Okumura ◽  
John P. Laurent ◽  
Marvin A. Fishman

✓ The effect of glycerol administration on cerebrospinal fluid (CSF) formation in dogs was studied by means of a ventriculocisternal perfusion technique. Net CSF production rate decreased after oral administration of glycerol (3 gm/kg) from a baseline level of 42.33 ± 6.68 µl/min (mean ± standard error) to a trough of 10.33 ± 4.88 µl/min at 90 minutes after administration (p < 0.025). Serum osmolality concomitantly increased from a baseline value of 296 ± 2.83 to 309 ± 4.7 mOsm/kg H2O at 90 minutes. The mean percentage change in CSF production inversely correlated to the mean percentage change in serum osmolality, r = −0.85. Thus, glycerol administration decreases net CSF formation, and this effect may be related in part to the rise in serum osmolality.


1974 ◽  
Vol 41 (3) ◽  
pp. 350-355 ◽  
Author(s):  
Michael E. Carey ◽  
A. Richard Vela

✓The rate of cerebrospinal fluid (CSF) production in dogs was measured by ventriculocisternal perfusion with artificial CSF containing inulin. In normotensive animals, the average CSF production was 36 ± 6 µl/min. When the mean arterial blood pressure was reduced to 62 ± 1 mm Hg, the CSF production fell to 22 ± 5 µl/min, a 39% reduction in fluid formation. The authors briefly discuss various hypotheses to explain this reduction.


1986 ◽  
Vol 251 (5) ◽  
pp. R996-R999
Author(s):  
A. A. Artru ◽  
T. F. Hornbein

When the rate of cerebrospinal fluid (CSF) production (Vf) is determined using the classical open ventriculocisternal perfusion technique, it is not possible to observe natural fluctuations in CSF pressure, or the effects of experimental treatments on CSF pressure, or to make inferences about CSF volume or resistance to reabsorption of CSF, because CSF pressure is fixed according to the level of the distal end of the cisternal outflow cannula. In addition, the convention of placing the distal end of the cisternal outflow cannula at the interaural line fixes CSF pressure at a value that may not be usual for the animal, thereby introducing potential causes for error in the determination of Vf. The present study describes and evaluates a method of closed ventriculocisternal perfusion that allows the simultaneous determination of Vf and CSF pressure. With this method the time to tracer equilibration was less, and the volume of distribution of the tracer was smaller than with the classical open perfusion system. CSF pressure tracings were of high fidelity showing both respiratory and cardiovascular variation. During both sedation with nitrous oxide or halothane anesthesia, Vf values using closed perfusion were similar to values previously reported using open perfusion. Vf decreased during halothane anesthesia compared with nitrous oxide sedation. No time-related change in Vf from the first measurable time period (approximately 2 h) to the end of the study (approximately 5 h) was observed.


2020 ◽  
Vol 65 (1) ◽  
pp. e01948-20
Author(s):  
Dalin Rifat ◽  
Si-Yang Li ◽  
Thomas Ioerger ◽  
Keshav Shah ◽  
Jean-Philippe Lanoix ◽  
...  

ABSTRACTThe nitroimidazole prodrugs delamanid and pretomanid comprise one of only two new antimicrobial classes approved to treat tuberculosis (TB) in 50 years. Prior in vitro studies suggest a relatively low barrier to nitroimidazole resistance in Mycobacterium tuberculosis, but clinical evidence is limited to date. We selected pretomanid-resistant M. tuberculosis mutants in two mouse models of TB using a range of pretomanid doses. The frequency of spontaneous resistance was approximately 10−5 CFU. Whole-genome sequencing of 161 resistant isolates from 47 mice revealed 99 unique mutations, of which 91% occurred in 1 of 5 genes previously associated with nitroimidazole activation and resistance, namely, fbiC (56%), fbiA (15%), ddn (12%), fgd (4%), and fbiB (4%). Nearly all mutations were unique to a single mouse and not previously identified. The remaining 9% of resistant mutants harbored mutations in Rv2983 (fbiD), a gene not previously associated with nitroimidazole resistance but recently shown to be a guanylyltransferase necessary for cofactor F420 synthesis. Most mutants exhibited high-level resistance to pretomanid and delamanid, although Rv2983 and fbiB mutants exhibited high-level pretomanid resistance but relatively small changes in delamanid susceptibility. Complementing an Rv2983 mutant with wild-type Rv2983 restored susceptibility to pretomanid and delamanid. By quantifying intracellular F420 and its precursor Fo in overexpressing and loss-of-function mutants, we provide further evidence that Rv2983 is necessary for F420 biosynthesis. Finally, Rv2983 mutants and other F420H2-deficient mutants displayed hypersusceptibility to some antibiotics and to concentrations of malachite green found in solid media used to isolate and propagate mycobacteria from clinical samples.


2015 ◽  
Vol 59 (5) ◽  
pp. 2867-2874 ◽  
Author(s):  
Atteneri López-Arencibia ◽  
Daniel García-Velázquez ◽  
Carmen M. Martín-Navarro ◽  
Ines Sifaoui ◽  
María Reyes-Batlle ◽  
...  

ABSTRACTThein vitroactivity of a novel group of compounds, hexaazatrinaphthylene derivatives, against two species ofLeishmaniais described in this study. These compounds showed a significant dose-dependent inhibition effect on the proliferation of the parasites, with 50% inhibitory concentrations (IC50s) ranging from 1.23 to 25.05 μM against the promastigote stage and 0.5 to 0.7 μM against intracellular amastigotes. Also, a cytotoxicity assay was carried out to in order to evaluate the possible toxic effects of these compounds. Moreover, different assays were performed to determine the type of cell death induced after incubation with these compounds. The obtained results highlight the potential use of hexaazatrinaphthylene derivatives againstLeishmaniaspecies, and further studies should be undertaken to establish them as novel leishmanicidal therapeutic agents.


2015 ◽  
Vol 197 (14) ◽  
pp. 2383-2391 ◽  
Author(s):  
Semen A. Leyn ◽  
Irina A. Rodionova ◽  
Xiaoqing Li ◽  
Dmitry A. Rodionov

ABSTRACTAutotrophic microorganisms are able to utilize carbon dioxide as their only carbon source, or, alternatively, many of them can grow heterotrophically on organics. Different variants of autotrophic pathways have been identified in various lineages of the phylumCrenarchaeota. Aerobic members of the orderSulfolobalesutilize the hydroxypropionate-hydroxybutyrate cycle (HHC) to fix inorganic carbon, whereas anaerobicThermoprotealesuse the dicarboxylate-hydroxybutyrate cycle (DHC). Knowledge of transcriptional regulation of autotrophic pathways inArchaeais limited. We applied a comparative genomics approach to predict novel autotrophic regulons in theCrenarchaeota. We report identification of two novel DNA motifs associated with the autotrophic pathway genes in theSulfolobales(HHC box) andThermoproteales(DHC box). Based on genome context evidence, the HHC box regulon was attributed to a novel transcription factor from the TrmB family named HhcR. Orthologs of HhcR are present in allSulfolobalesgenomes but were not found in other lineages. A predicted HHC box regulatory motif was confirmed byin vitrobinding assays with the recombinant HhcR protein fromMetallosphaera yellowstonensis. For the DHC box regulon, we assigned a different potential regulator, named DhcR, which is restricted to the orderThermoproteales. DhcR inThermoproteus neutrophilus(Tneu_0751) was previously identified as a DNA-binding protein with high affinity for the promoter regions of two autotrophic operons. The global HhcR and DhcR regulons reconstructed by comparative genomics were reconciled with available omics data inMetallosphaeraandThermoproteusspp. The identified regulons constitute two novel mechanisms for transcriptional control of autotrophic pathways in theCrenarchaeota.IMPORTANCELittle is known about transcriptional regulation of carbon dioxide fixation pathways inArchaea. We previously applied the comparative genomics approach for reconstruction of DtxR family regulons in diverse lineages ofArchaea. Here, we utilize similar computational approaches to identify novel regulatory motifs for genes that are autotrophically induced in microorganisms from two lineages ofCrenarchaeotaand to reconstruct the respective regulons. The predicted novel regulons in archaeal genomes control the majority of autotrophic pathway genes and also other carbon and energy metabolism genes. The HhcR regulon was experimentally validated by DNA-binding assays inMetallosphaeraspp. Novel regulons described for the first time in this work provide a basis for understanding the mechanisms of transcriptional regulation of autotrophic pathways inArchaea.


2011 ◽  
Vol 56 (1) ◽  
pp. 148-153 ◽  
Author(s):  
Marisa H. Miceli ◽  
Stella M. Bernardo ◽  
T. S. Neil Ku ◽  
Carla Walraven ◽  
Samuel A. Lee

ABSTRACTInfections and thromboses are the most common complications associated with central venous catheters. Suggested strategies for prevention and management of these complications include the use of heparin-coated catheters, heparin locks, and antimicrobial lock therapy. However, the effects of heparin onCandida albicansbiofilms and planktonic cells have not been previously studied. Therefore, we sought to determine thein vitroeffect of a heparin sodium preparation (HP) on biofilms and planktonic cells ofC. albicans. Because HP contains two preservatives, methyl paraben (MP) and propyl paraben (PP), these compounds and heparin sodium without preservatives (Pure-H) were also tested individually. The metabolic activity of the mature biofilm after treatment was assessed using XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] reduction and microscopy. Pure-H, MP, and PP caused up to 75, 85, and 60% reductions of metabolic activity of the mature preformedC. albicansbiofilms, respectively. Maximal efficacy against the mature biofilm was observed with HP (up to 90%) compared to the individual compounds (P< 0.0001). Pure-H, MP, and PP each inhibitedC. albicansbiofilm formation up to 90%. A complete inhibition of biofilm formation was observed with HP at 5,000 U/ml and higher. When tested against planktonic cells, each compound inhibited growth in a dose-dependent manner. These data indicated that HP, MP, PP, and Pure-H havein vitroantifungal activity againstC. albicansmature biofilms, formation of biofilms, and planktonic cells. Investigation of high-dose heparin-based strategies (e.g., heparin locks) in combination with traditional antifungal agents for the treatment and/or prevention ofC. albicansbiofilms is warranted.


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