Comparison of the in vitro activities of delafloxacin and comparators against Staphylococcus epidermidis clinical strains involved in osteoarticular infections: a CRIOGO multicentre retrospective study.

OBJECTIVES: Staphylococcus epidermidis bone and joint infections (BJIs) on material are often difficult to treat. The activity of delafloxacin has not yet been studied on S. epidermidis in this context. The aim of this study was to assess its in vitro activity compared with other fluoroquinolones, against a large collection of S. epidermidis clinical strains. METHODS: We selected 538 S. epidermidis strains isolated between January 2015 and February 2023 from six French teaching hospitals. One hundred and fifty-two strains were ofloxacin susceptible and 386 were ofloxacin resistant. Identifications were performed by MS and MICs were determined using gradient concentration strips for ofloxacin, levofloxacin, moxifloxacin and delafloxacin. RESULTS: Ofloxacin-susceptible strains were susceptible to all fluoroquinolones. Resistant strains had higher MICs of all fluoroquinolones. Strains resistant to ofloxacin (89.1%) still showed susceptibility to delafloxacin when using the Staphylococcus aureus 2021 CA-SFM/EUCAST threshold of 0.25 mg/L. In contrast, only 3.9% of the ofloxacin-resistant strains remained susceptible to delafloxacin with the 0.016 mg/L S. aureus breakpoint according to CA-SFM/EUCAST guidelines in 2022. The MIC50 was 0.094 mg/L and the MIC90 was 0.38 mg/L. CONCLUSIONS: We showed low delafloxacin MICs for ofloxacin-susceptible S. epidermidis strains and a double population for ofloxacin-resistant strains. Despite the absence of breakpoints for S. epidermidis, delafloxacin may be an option for the treatment of complex BJI, including strains with MICs of ≤0.094 mg/L, leading to 64% susceptibility. This study underlines the importance for determining specific S. epidermidis delafloxacin breakpoints for the management of BJI on material.

Effects of chronic insomnia and use of benzodiazepines on daytime performance in older adults.

This study evaluated the impact of insomnia and chronic use of benzodiazepines on the cognitive and psychomotor performance of older adults. Three conditions, matched on age, gender, and education, were compared: 20 prolonged users of benzodiazepines for insomnia, 20 unmedicated insomniacs, and 20 good sleepers. The participants completed neuropsychological tests of memory, attention/concentration, psychomotor speed, and executive functions, as well as subjective evaluations of their actual performance. Individuals with insomnia, both medicated and unmedicated, performed worse than good sleepers on the attention/concentration factor. There was no other objective evidence of performance impairments. However, unmedicated insomniacs had lower performance expectancies and subjectively rated their performance more negatively relative to medicated insomniacs and good sleepers. Both insomnia conditions also rated their performance as lower compared with their perceived potential. It is suggested that the attention/concentration difficulties experienced by medicated and unmedicated older adults with insomnia may be linked to a state of hyperarousal. The discrepancies between subjective reports of daytime deficits and objective impairments may reflect a generalized faulty appraisal of sleep and daytime functioning among individuals with insomnia complaints. The implications of those findings for the assessment and treatment of late-life insomnia are discussed.

Pleomorphic soft tissue myogenic sarcomas of adulthood. A reappraisal in the mid-1990s.

325 diverse sarcomas, 39 rhabdomyosarcomas (RMS), including all histologic variants, and 135 leiomyosarcomas (LMS) were identified. Within these two groups, 18 (46%) of the RMS and 14 (10%) of the LMS represented pleomorphic variants. These neoplasms were studied by morphology (histology and ultrastructure) and by immunohistochemical methods employing antibodies to intermediate filaments (vimentin and desmin) and actin isoforms [alpha-smooth (sm) and alpha-sarcomeric (sr) actins]. Twenty-four pleomorphic malignant fibrous histiocytomas (MFH) and eight pleomorphic liposarcomas (LS) were examined in a similar fashion. By light microscopy, the pleomorphic RMS, LMS, and MFH were indistinguishable, as each was dominated by pleomorphic cells disposed in a haphazard growth pattern; moreover, many featured fascicular, storiform, and sclerotic zones. The distinction between these neoplasms became apparent only following immunohistochemistry and/or ultrastructural study. All pleomorphic RMS disclosed rudimentary sarcomeres and exhibited the following cytoskeletal profile: vimentin (+) (18 of 18), desmin (+) (14 of 18), alpha-sr actin (+) (18 of 18) and alpha-sm actin (+) (five of 18). All the pleomorphic LMS featured smooth-muscle differentiation of variable degrees in the form of cytoplasmic bundles of microfilaments and associated dense bodies; their cytoskeletal profile was vimentin (+) (14 of 14), desmin (+) (seven of 14), alpha-sr actin (+) (none of 14), and alpha-sm actin (+) (eight of 14). The latter was demonstrated in all moderately differentiated, but absent or only focally expressed in poorly differentiated variants. All pleomorphic MFH and LS were devoid of myogenic (skeletal or smooth) ultrastructural features and expressed vimentin solely. This combined morphological and immunohistochemical study illustrates the following: First, these pleomorphic sarcomas are often indistinguishable by histologic growth pattern alone; thus, an accurate diagnosis requires study with all of these techniques. Second, pleomorphic myogenic sarcomas are restricted to adults and are not uncommon neoplasms among pleomorphic sarcomas: RMS (28%), LMS (21%), MFH (38%), and LS (13%). Third, the study defines desmin-negative and alpha-sm actin-positive pleomorphic RMS, and desmin-negative and alpha-sm-actin-negative pleomorphic LMS.

A reproducible method for identification of human genomic DNA autonomously replicating sequences.

We demonstrate a method for the isolation of autonomously replicating sequences from pools of clones obtained from genomic DNA libraries constructed using affinity purification of cruciform DNA. The selection of autonomously replicating sequences was based on their differential ability to replicate as episomes after transfection of pools of plasmid clones into human HeLa cells. Two separate libraries containing affinity-purified cruciform DNA were used, one prepared from DNA of log phase primary human genital fibroblasts and the other prepared from DNA of log phase SW48 colon adenocarcinoma cells. Representative samples of the entire phage libraries were converted to phagemid clones by filamentous helper phage-mediated mass excision to produce pBluescript libraries in Escherichia coli. Clones were grown up individually and the bacteria pooled into groups of 48 for recovery of plasmid DNA. Plasmid pools of 48 independent clones (120 micrograms total) were then transfected by calcium phosphate coprecipitation onto log phase HeLa cells, which were allowed to grow for 3 days before recovery of plasmid by Hirt lysis. The recovery of plasmid from each transfection was estimated to range from 10 to 60 ng. DpnI digestion was then used to digest plasmids which had not been replicated and therefore retained a bacterial methylation pattern which was sensitive to digestion. We estimated from agarose electrophoresis gels that 40-200 pg of recovered plasmid DNA per transfected pool of DNA was resistant to DpnI and therefore was capable of transforming competent E. coli cells. The DpnI-resistant fraction yielded from one to seven independent clones from each pool, with genomic DNA inserts ranging in size from 0.35 to 3.4 kb.(ABSTRACT TRUNCATED AT 250 WORDS)

cDNA clones contain autonomous replication activity.

We have undertaken to investigate transcription as a regulatory event in mammalian DNA replication. Subpopulations of transcripts represented in a cDNA library of human embryo lung fibroblasts (IMR90) were examined for their ability to support autonomous replication after transfection into human cells (HeLa). Two of three cDNA clones (343, 363) containing 'O'-family repetitive sequences, after subcloning into pBR322 and transfection into HeLa cells, were capable of autonomous replication. One of these cDNA clones, 343, is enriched by selection for poly(A)+ RNA. In contrast, none of five Alu-containing transcripts was capable of autonomous replication in human cells. However, six out of ten cDNA clones contained neither 'O'-family or Alu homologous sequences and were as efficient as the cDNA clones containing 'O'-family sequences in replicating autonomously in human cells. cDNA clones, from an oligo-d(T)-primed library of human poly(A)+ enriched RNA, contain a significant proportion of independent clones that can also support autonomous replication of bacterial plasmids in human cells. cDNA clone 343 was observed to contain in a 448 bp EcoRI-HincII fragment, yeast ARS consensus, SAR consensus, IRs, bent DNA and a DUE, all sequence and structural characteristics often associated with many prokaryotic, viral and eukaryotic origins. Sequence analysis of seven other cDNA clones (from non-'O'-family, non-Alu homologous sequences, NOA) showed that five contained some of the same consensus sequences. Two NOA clones (NOA4 and -5) did not contain any representations of ARS and SAR consensus sequences, suggesting that these two features may not be essential for autonomous replication activity in mammalian cells.

Effect of congestive heart failure on clentiazem pharmacokinetics in a dog model.

Clentiazem, 8-chloro diltiazem, is a calcium channel blocker currently undergoing evaluation for the treatment of stable angina and hypertension. As patients with ischaemic disorders often present some degree of heart failure, the aim of this study was to investigate the effect of congestive heart failure on clentiazem (200 micrograms kg-1, i.v. bolus) pharmacokinetics in a canine model. Congestive heart failure was induced in six dogs by rapid ventricular pacing (240 beats min-1) for 3-5 weeks. Clentiazem pharmacokinetics was studied in each dog under the control condition and after the development of clinical signs of heart failure (ascites, dyspnea, fatigue). Blood samples were collected up to 480 min post-dose. Clentiazem plasma concentrations were determined by high performance liquid chromatography. The area under the plasma concentration versus time curves (AUC0-infinity) was significantly increased in congestive heart failure dogs (8.8 +/- 1.6 vs 21.8 +/- 1.4 micrograms min ml-1) (mean +/- SEM). These changes were related to a reduction of the volume of distribution of the central compartment (0.9 +/- 0.1 vs 0.2 +/- 0.11 kg-1) and total body clearance (1.9 +/- 0.4 vs 0.7 +/- 0.21 h-1 kg-1). It is concluded that, in our model, congestive heart failure significantly modifies clentiazem disposition. These results suggest that caution should be exercised when clentiazem is given to patients with a low ejection fraction and a compromised cardiac function. Reduced loading and maintenance doses might be recommended in patients with severe congestive heart failure.

Lactose conjugation of sulphonamide drugs in the lactating dairy cow.

1. 14C-Sulphamethazine (4-amino-N-(4,6-dimethyl-2-pyrimidinyl)benzene-[U-14C]-sulphonamide; 220 mg/kg of body weight) was given orally or i.v. to lactating dairy cows. Milk collected from 0-48 h after dosing accounted for 2.0% (oral dose) and 1.1% (i.v. dose) of the total 14C-activity administered. 2. Sulphamethazine accounted for 70-79% (oral dose) and 54-75% (i.v. dose) of the total 14C in milk samples collected from 0-48 h after dosing. N4-acetylsulphamethazine accounted for 1-2% (oral dose) and 1-4% (i.v. dose) of the 14C in milk. 3. The major 14C-labelled metabolite in the milk was isolated and identified as the N4-lactose conjugate of sulphamethazine, a unique type of metabolite not previously reported. This metabolite accounted for 10-14% (oral dose) and 9-20% (i.v. dose) of the 14C-activity in the milk collected from 0-48 h after dosing with 14C-sulphamethazine. 4. N4-lactose conjugates of sulphapyridine, sulphamerazine, sulphathiazole, sulphadimethoxine and sulphaquinoxaline were present in the milk from cows orally dosed with these five sulphonamide drugs.

Dose-dependent cytotoxicity of chlorinated hydrocarbons in isolated rat hepatocytes.

The aim was to determine if isolated suspended hepatocytes could differentiate between the effects of four chlorinated hydrocarbons that are hepatotoxic in vivo and four that are not. Membrane integrity was assessed by measuring alanine aminotransferase (ALT) release after 30- to 180-min incubations in vitro. From the results, the chlorinated hydrocarbons fell into three groups: tetrachloroethylene and 1,1,2,2-tetrachloroethane were the most potent cytotoxicants; CCl4, 1,1,2-trichloroethane, and trichloroethylene exhibited intermediate cytotoxicity; and low cytotoxicity was observed with CHCl3, 1,1,1-trichloroethane, and 1,1-dichloroethylene. Cytotoxicity ranking correlated poorly with the reported in vivo hepatotoxicity of these agents. The effect of adding SKF-525A on the cytotoxicity of tetrachloroethylene and CCl4 was also assessed. In addition, hepatocytes from rats pretreated with 2,5-hexanedione were used to determine if they were more susceptible to the effects of CHCl3, CCl4, or tetrachloroethylene. SKF-525A decreased the cytotoxicity of both CCl4 and tetrachloroethylene, whereas pretreatment with 2,5-hexanedione enhanced their effect. The effects of both SKF-525A and 2,5-hexanedione on CCl4 in vitro are consistent with in vivo findings. However, tetrachloroethylene is not hepatotoxic in vivo, suggesting that SKF-525A might act by stabilizing plasma membranes rendering the hepatocyte more resistant to lysis. Overall, the results cast doubts on the use of ALT release from isolated hepatocytes as an appropriate in vitro model for assessing hepatotoxic properties of chlorinated hydrocarbons.

Comparison of 252Cf plasma desorption mass spectrometry and fast atom bombardment mass spectrometry in identification of simple monoglucosyl conjugates.

The fast atom bombardment (glycerol) and 252Cf plasma desorption mass spectra of 16 simple monoglucosyl conjugates have been compared. Plasma desorption mass spectrometry, in general, has been found to be the superior technique for characterizing these low molecular weight conjugates because of the relative absence of interfering matrix peaks.

Interaction between prostaglandin E2 and leukotriene D4 on the excretion of electrolytes by the dog kidney in vivo.

Recent experiments indicate that prostaglandin E2 potentiates the vasodilatory properties of leukotrienes in the skin microcirculation. The present experiments were undertaken to study the effect of leukotriene D4 and prostaglandin E2 on renal hemodynamics and urinary electrolytes in the dog. Experiments were performed in three groups of anesthetized Mongrel dogs: the first group was studied under hydropenia, whereas the two remaining groups were studied during water diuresis with (Group 3) or without indomethacin (Group 2). LTD4 (100 ng/min) and PGE2 (3 ug/min) were infused in the left renal artery to minimize systemic effects of these compounds. LTD4 alone failed to influence urinary sodium excretion in all 3 groups. In Group 1, urinary sodium increased from 77 +/- 6 to 393 +/- 74 uEq/min during PGE2, and further increased to 511 +/- 52 uEq/min during LTD4 + PGE2. No change occurred in the contralateral right kidney. In this group, glomerular filtration as well as renal plasma flow were not statistically influenced. In Group 2, the same phenomenon was observed for urinary sodium. The combined infusion of LTD4 + PGE2 increased urinary sodium without significant changes in glomerular filtration and renal plasma flow. Finally, in Group 3, indomethacin was shown to reduce the natriuretic effects of LTD4 and PGE2: during PGE2 alone, urinary sodium increased from 90 +/- 14 to 260 +/- 66 uEq/min, and only rose from 80 +/- 10 to 175 +/- 19 uEq/min during the combined infusion of LTD4 and PGE2. In groups 2 and 3, free water clearance was utilized as an index of sodium chloride reabsorption in the thick ascending limb: this parameter increased from 2.35 +/- 0.25 to 4.70 +/- 0.30 ml/min, while urinary volume was increasing from 3.55 +/- 0.25 to 10.05 +/- 0.65 ml/min, during LTD4 + PGE2. Indomethacin, administered in Group 3, (3 mg/kg/hr) again abolished the effect of combined PGE2 + LTD4. These results indicate a potentiating effect of leukotriene D4 on the PGE2-induced natriuresis in the anesthetized dog. These phenomena occurred in the absence of significant changes in renal hemodynamics, therefore suggesting a direct tubular effect of these arachidonic acid metabolites. Finally, the water diuresis experiments suggest a proximal site of action of PGE2 and LTD4.

Hemodynamic effects of PAF-acether.

Intravenous PAF-Acether produces extravasation of plasma and systemic hypotension. Blood flow to vital organs, including the kidneys, is markedly reduced as a consequence. This study examines the role of cyclo-oxygenase metabolites and angiotensin II in mediating the hemodynamic effects and the renal consequences of PAF-Acether injection. Indomethacin prevents the reduction of arterial blood pressure during PAF infusion. However, the fall in glomerular filtration and renal plasma flow is not abolished by this treatment: inulin and PAH clearances fall from 42 +/- 2 to 30 +/- 1 ml/min, and from 102 +/- 5 to 59 +/- 4 ml/min, respectively. Similarly, indomethacin does not prevent the fall in urinary sodium excretion. However, when angiotensin II receptors blockade is added to indomethacin, glomerular filtration is not statistically affected during PAF infusion. PAH clearance only decreases from 100 +/- 8 to 87 +/- 8 ml/min. However, the combined administration of indomethacin and saralasin does not prevent the fall in urinary sodium excretion, which decreases from 132 +/- 10 to 67 +/- 6 mu Eq/min. The results therefore indicate that the vascular effects of PAF-acether are heterogeneous: the peripheral actions require the production of vasodilatory prostaglandins, whereas in the kidney, inhibition of prostaglandins synthesis does not prevent the effect on renal hemodynamics and sodium excretion. It is likely that the vasoconstrictor effect of PAF on renal plasma flow and glomerular filtration results from increased release of angiotensin II, since saralasin prevents those effects. The mechanisms responsible for the dissociation between renal hemodynamics and urinary sodium excretion require further studies.

Potentiating effects of leukotriene B4 and prostaglandin E2 on urinary sodium excretion by the dog kidney.

A synergistic vasodilatation was recently demonstrated with leukotriene B4 (LTB4) and prostaglandin E2 (PGE2) in the cutaneous microcirculation. The present study addresses this question to the renal microcirculation, with respect to its eventual influence on the net transport of sodium, infusing small doses of LTB4 (100 ng/min) and PGE2 (3 ng/min) in the left renal artery, and using the contralateral kidney as control in anesthetized Mongrel dogs. In group 1 (hydropenic animals), LTB4 alone failed to influence natriuresis (UNaV) while PGE2 increased UNaV from 152 +/- 20 to 225 +/- 18 uEq/min. The combined infusion of LTB4 and PGE2 resulted in a marked elevation of natriuresis to 368 +/- 26, 317 +/- 30 and 342 +/- 52 uEq/min. In group 2, water diuresis was induced to examine the eventual site of action of these compounds, and to assess the response of the diluting segment of the nephron. In these dogs, PGE2 was first administered and UNaV rose modestly from 51 +/- 12 to 77 +/- 15 uEq/min. LTB4 again had no significant influence on UNaV, but LTB4 and PGE2 produced a marked increment from 75 +/- 16 to 374 +/- 24 uEq/min. Urine volume, as well as free water clearance, increased from 3.3 +/- 0.2 to 6.5 +/- 0.7 ml/min, and from 2.3 +/- 0.2 to 3.7 +/- 0.4 ml/min, respectively, during LTB4 + PGE2. No significant change occurred in the right control kidney during these manoeuvers. Since renal hemodynamics (glomerular filtration and plasma flow) remained relatively stable in both groups of dogs, it is suggested that the combined infusion of LTB4 and PGE2 exerts a direct influence on the net transport of sodium, probably in the proximal tubule, as inferred by the results obtained in group 2. These two important metabolites of arachidonic acid could be involved in the modulation of renal sodium excretion under normal and/or pathophysiological conditions.

Hypotension and arteriovenous shunting: effects of intravenous infusion of nitroprusside, nitroglycerin and phentolamine.

The effects of nitroprusside, nitroglycerin and phentolamine on cardiac dynamics and on the fraction of cardiac output shunted through systemic arteriovenous communications, which may explain disparate responses elicited by these systemic vasodilators upon venous return, have been studied in 15 nonanesthetized dogs. Cardiac dynamic parameters were measured by electromagnetic flow probe placed at the root of the aorta. Quantitative measurements of total systemic arteriovenous shunting were determined from the fraction of 9 mu radioactively labeled microspheres, injected into the left atrium, recovered in the pulmonary artery. To provide a common basis for comparison, the mean arterial pressure was lowered by 15-20% either with an intravenous infusion of nitroprusside, nitroglycerin or phentolamine. At the fifteenth minute of infusion, nitroprusside produced significant decrease in stroke volume index (23%) and left ventricular power and work (28% and 40%). Nitroglycerin decreased significantly stroke volume index (12%), cardiac index (9%) and left ventricular work (22%). Phentolamine significantly increased heart rate (72%) and left ventricular maximum acceleration (30%) while it decreased stroke volume index (41%), left ventricular power and work (19% and 55%). Total peripheral resistance was significantly affected only by infusion of phentolamine (-18%). Left ventricular maximum velocity, mean systolic ejection rate and maximum systolic flow did not change significantly under infusion of these systemic vasodilators. Under control conditions, total systemic shunting of cardiac output averaged 8.9-10% and was not modified by any of the vasodilators used. Arteriovenous O2 difference and oxygen consumption, corroborated these findings since they remained within normal limits before and after infusion of nitroprusside, nitroglycerin or phentolamine.(ABSTRACT TRUNCATED AT 250 WORDS)

Nephrogenic adenoma of the bladder after kidney transplantation: spontaneous improvement with azathioprine removal; surgical trauma and cytomegalovirus infection as possible etiologic factors.

We report 2 cases of nephrogenic adenoma several years after successful cadaver kidney transplantation. In 1 case the lesion had cytomegalovirus inclusions, and we observed a marked and sustained reduction in the extension of the lesion with cessation of azathioprine. Surgical trauma and cytomegalovirus infections are discussed as etiologic factors. Conservative treatment seemed appropriate.

AIDS in Haitian immigrants and in a Caucasian woman closely associated with Haitians.

In Montreal the acquired immune deficiency syndrome (AIDS) was seen in eight Haitian immigrants and one Caucasian woman who had lived with Haitian immigrants for 3 years before the onset of her illness. AIDS was characterized by opportunistic infections alone in seven patients, by opportunistic infection and Kaposi's sarcoma in one patient and by chronic generalized lymphadenopathy in one patient. Five of the patients had presented with Mycobacterium tuberculosis infections 1 to 12 months before the onset of opportunistic infections. All nine patients were found to have recall anergy by skin testing for delayed hypersensitivity. Enumeration of the lymphocyte subpopulations in three patients showed a marked inversion of the ratio of helper to suppressor T lymphocytes. Six of the patients died as a result of the opportunistic infections; autopsies showed no recognizable causes of immunodeficiency. Thus, there is in Montreal a third clustering of AIDS cases in North America related to Haitian immigrants.