Thermodynamics of the structural transition in metal-organic frameworks.

A thermodynamic study of the structural large-pore (LP) to narrow pore (NP) transition in various Metal Organic Frameworks (MOFs) is presented. First, the pressure induced transition at a constant temperature is investigated using a Tian-Calvet microcalorimeter set-up equipped with a high pressure cell. This device permits simultaneous measurements of the mechanical work and heat associated with the LP → NP transition. It is shown that MIL-53(Al) and MIL-53(Cr) have similar thermodynamic and mechanical behaviour whilst the MIL-47(V) system is characterized by much higher transition energy and mechanical work. Second, the temperature induced transition at ambient pressure is studied by means of differential scanning calorimetry (DSC) combined with X-ray absorption spectroscopy. This set-up enables one to follow simultaneously the structural changes associated with the phase transition detected by DSC. The MIL-53(Cr)-Br functionalized MOF is chosen here as a case study where both energetics and structural changes are discussed.

Pharmacodynamics of ibafloxacin in micro-organisms isolated from cats.

The pharmacodynamic properties of ibafloxacin were investigated in micro-organisms isolated from cats. Minimal inhibitory concentrations (MIC) of ibafloxacin (racemate, R- and S-enantiomers) and its metabolites (7-hydroxy- and 8-hydroxy-ibafloxacin) and time-kill kinetics were determined against Gram-negative and Gram-positive bacteria isolated from dermal and respiratory and urinary tract infections in cats. Racemic ibafloxacin has a broad spectrum of bactericidal activity against Gram-negative and some Gram-positive bacteria. Escherichia coli and Pasteurella, Klebsiella and Staphylococcus spp. are commonly isolated from feline infections and all are susceptible to ibafloxacin (MIC90 < or = 0.5 microg/mL), whereas Pseudomonas aeruginosa, Proteus mirabilis and Streptococcus spp. are considered intrinsic resistant. Microbiological activity resides primarily in the S-enantiomer of ibafloxacin whereas the R-enantiomer is less active. Killing curves using concentrations of racemic ibafloxacin and 8-hydroxy-ibafloxacin, which are representative of the in vivo situation observed in cats, showed at least 99.9% reduction in viable bacterial isolates from feline clinical samples over 24 h. Bacterial eradication was achieved in cats with Cmax/MIC and AUC/MIC values much lower than the target values previously established in man and laboratory animals. Additional studies in dogs and cats are necessary to define more clearly the surrogate markers of antibacterial activity (i.e. Cmax/MIC, AUC/MIC ratios), which are associated with a good clinical response.

Pharmacokinetics of ibafloxacin in healthy cats.

The pharmacokinetics of ibafloxacin following single and repeated administration of an oral gel formulation and the effect of food intake were investigated in cats. Ibafloxacin is a chiral fluoroquinolone available for clinical use as a racemic mixture of the R- and S-enantiomers. Plasma concentrations of ibafloxacin and its metabolites were determined using microbiological, LC-MS-MS and enantioselective capillary zone electrophoresis assays. Ibafloxacin was absorbed rapidly [time of maximum concentration (tmax) 2-3 h], reaching a mean maximum concentration (Cmax) of approximately 2.1 and 1.6 microg/mL for R- and S-ibafloxacin, respectively, following a single oral administration of the racemate at 15 mg/kg. Once absorbed, ibafloxacin was metabolized to 7-hydroxy-ibafloxacin and mainly to 8-hydroxy-ibafloxacin. Following repeated oral administration, significant increases in Cmax and AUC of ibafloxacin and its less active metabolites (racemic or enantiomers) were observed between the first and the tenth day of treatment. This twofold exposure increase in concentrations of ibafloxacin and its metabolites may contribute additionally to the efficacy of this drug in the treatment of feline bacterial infections. Single and repeated doses of ibafloxacin were well tolerated by cats. Food promoted the absorption of ibafloxacin, doubling Cmax and increasing AUC and slightly delaying tmax. High concentrations of the metabolites, mainly 8-hydroxy- and 7-hydroxy-ibafloxacin were excreted in urine, either unchanged or as glucurono-conjugates.

In vitro and in vivo pharmacodynamic properties of the fluoroquinolone ibafloxacin.

The pharmacodynamic properties of a new veterinary fluoroquinolone antimicrobial agent, ibafloxacin, were evaluated. Minimal inhibitory concentrations (MIC), time-kill kinetics, postantibiotic effect (PAE) and postantibiotic subminimal inhibitory concentration effects (PA-SME) were determined against pathogenic canine Gram-negative and Gram-positive bacterial isolates from dermal, respiratory and urinary tract infections. The synergistic interactions between ibafloxacin and its main metabolite, 8-hydroxy-ibafloxacin were investigated. Finally, the efficacy of ibafloxacin was tested in in vivo canine infection models. Ibafloxacin had good activity against Pasteurella spp., Escherichia coli, Klebsiella spp., Proteus spp. and Staphylococcus spp. (MIC90=0.5 microg/mL), moderate activity against Bordetella bronchiseptica, Enterobacter spp. and Enterococcus spp. (MIC50=4 microg/mL) and low activity against Pseudomonas spp. and Streptococcus spp. The time-killing analysis confirmed that ibafloxacin was bactericidal with a broad spectrum of activity. The PAE and PA-SME were between 0.7-2.13 and 1-11.5 h, respectively. Finally, studies in dog models of wound infection and cystitis confirmed the efficacy of once daily oral ibafloxacin at a dosage of 15 mg/kg. Additional studies are needed to better define the importance of AUC/MIC (AUIC) and Cmax/MIC ratios on the outcome of fluoroquinolone therapy in dogs.

Pharmacokinetics of ibafloxacin following intravenous and oral administration to healthy Beagle dogs.

The pharmacokinetics of ibafloxacin, a new veterinary fluoroquinolone antimicrobial agent, was studied following intravenous (i.v.) and oral administration to healthy dogs. The mean absolute bioavailability of ibafloxacin after oral doses of 7.5, 15 and 30 mg/kg ranged from 69 to 81%, indicating that ibafloxacin was well absorbed by dogs. Ibafloxacin was also absorbed rapidly [time of maximum concentration (t(max)) 1.5 h], reaching a mean maximum concentration (C(max)) of 6 microg/mL at 15 mg/kg, well distributed in the body [large volume of distribution at steady state (V(ss)) and V(area) of 1.1 L/kg and 4 L/kg, respectively], and exhibited an elimination half-life of 5.2 h and a low total body clearance (8.7 mL/min/kg). Both C(max) and area under the concentration-time curve (AUC) showed dose proportionality over the dose range tested (7.5-30 mg/kg). The pharmacokinetics of ibafloxacin was similar following single and repeated dosage regimens, implying no significant accumulation in plasma. Food promoted the absorption of ibafloxacin by increasing C(max) and AUC, but did not change t(max). High amounts of the metabolites, mainly 8-hydroxy- and, 7-hydroxy-ibafloxacin were excreted in urine and faeces, either unchanged or as glucuronide conjugates. Following oral administration of 15 mg ibafloxacin/kg, the total recovery of ibafloxacin, its metabolites and conjugates in urine and faeces was 61.9-99.9% of the dose within 48 h.

Evidence for cytochrome P4501A2-mediated protein covalent binding of thiabendazole and for its passive intestinal transport: use of human and rabbit derived cells.

Thiabendazole (TBZ), an anthelmintic and fungicide benzimidazole, was recently demonstrated to be extensively metabolized by cytochrome P450 (CYP) 1A2 in man and rabbit, yielding 5-hydroxythiabendazole (5OH-TBZ), the major metabolite furtherly conjugated, and two minor unidentified metabolites (M1 and M2). In this study, exposure of rabbit and human cells to 14C-TBZ was also shown to be associated with the appearance of radioactivity irreversibly bound to proteins. The nature of CYP isoforms involved in this covalent binding was investigated by using cultured rabbit hepatocytes treated or not with various CYP inducers (CYP1A1/2 by beta-naphthoflavone, CYP2B4 by phenobarbital, CYP3A6 by rifampicine, CYP4A by clofibrate) and human liver and bronchial CYP-expressing cells. The covalent binding to proteins was particularly increased in beta-naphthoflavone-treated rabbit cells (2- to 4-fold over control) and human cells expressing CYP1A2 (22- to 42-fold over control). Thus, CYP1A2 is a major isoenzyme involved in the formation of TBZ-derived residues bound to protein. Furthermore, according to the good correlation between covalent binding and M1 or 5OH-TBZ production, TBZ would be firstly metabolized to 5OH-TBZ and subsequently converted to a chemically reactive metabolic intermediate binding to proteins. This metabolic activation could take place preferentially in liver and lung, the main biotransformation organs, rather than in intestines where TBZ was shown to be not metabolized. Moreover, TBZ was rapidly transported by passive diffusion through the human intestinal cells by comparison with the protein-bound residues which were not able to cross the intestinal barrier. Consequently, the absence of toxicity measured in intestines could be related to the low degree of TBZ metabolism and the lack of absorption of protein adducts. Nevertheless, caution is necessary in the use of TBZ concurrently with other drugs able to regulate CYP1A2, particularly in respect to liver and lung tissues, recognised as sites of covalent-binding.

Identification of human and rabbit cytochromes P450 1A2 as major isoforms involved in thiabendazole 5-hydroxylation.

This report characterized one of the major cytochrome P450 isozyme involved in thiabendazole metabolism. This study was undertaken by using both cultured rabbit hepatocytes treated or not with drugs known to specifically induced various cytochromes P450 isoenzymes (i.e., P450 1A1/2 by beta-naphthoflavone, P450 2B4 by phenobarbital, P450 3A6 by rifampicine and P450 4A by clofibrate) and human liver (THLE-5) and bronchial (BEAS-2B) epithelial cells expressing or not the major constitutive human cytochromes P450 (i.e., CYP1A2, 2A6, 2B6, 2C9, 2D6, 2E1 or 3A4). Only hepatocytes exposed to beta-naphthoflavone and clofibrate significantly metabolized thiabendazole to 5-hydroxythiabendazole. Extensive biotransformation of this anthelmintic only occurred in human cells expressing CYP1A2. Moreover, experiments performed on rabbit preparations showed good correlations between thiabendazole 5-hydroxylase activity and both ethoxyresorufin and methoxyresorufin O-dealkylase activities. Thus, CYP1A2 is a major isoenzyme involved in thiabendazole 5-hydroxylation.

Comparative Metabolism of Thiabendazole in Cultured Hepatocytes from Rats, Rabbits, Calves, Pigs, and Sheep, Including the Formation of Protein-Bound Residues.

Cultured hepatocytes from rat, rabbit, calf, pig, and sheep were used to study metabolism and formation of protein-bound residues of thiabendazole ([(14)C]TBZ), a benzimidazole anthelmintic and fungicide. In all investigated species, major pathways corresponded to 5-hydroxylation of TBZ and its further conjugation. However, marked interspecies differences in rates of TBZ disappearance and 5-hydroxy metabolite formation were demonstrated. Rabbit hepatocytes presented the fastest TBZ biotransformation and were the most extensive hydroxylators. By contrast, the lowest capacity of oxidation was observed for the rat. Two unidentified minor metabolites, designated M1 and M2, were particularly produced by sheep hepatocytes. Moreover, the protein-bound residues in these cells, which could be related to cytochrome P450-dependent oxidation, were formed in 4 times greater amounts than in the other animal cells. These findings substantiate hepatocytes as an in vitro model for prediction of hepatic metabolism in vivo.

Ontogenic development of drug-metabolizing enzymes in male chicken liver.

The ontogenic study of the hepatic biotransformation enzymes revealed the early development of both oxidative and conjugative enzymes in male chickens ranging in age from 3 to 12 weeks. Although the rate of microsomal cytochrome P450 reactions progressively increased during the first 9 weeks, it decreased thereafter. Furthermore, the proteins revealed by the antibodies to anti-cytochrome P450 1A2, 2B4, 2C7, and 3A4 appeared to be constitutively expressed. Hepatic monooxygenases were characterized by different developmental patterns. The demethylase activities increased progressively up to 9 weeks, then they declined, in 12 weeks reaching the activity level observed in 3-week-old chickens. In contrast, alkoxyresorufin O-dealkylases and benzopyrene hydroxylase activities continued to increase with age. Significant variability was noted for aniline hydroxylase. Among conjugation enzymes, UDP-glucuronyltransferase towards p-nitrophenol and isoniazid N-acetyltransferase activities increased with the age of the fowl, but with different profiles. Concerning glutathione S-transferase accepting 1-chloro-2,4-dinitrobenzene or 1,2-dichloro-4-nitrobenzene, the chickens aged from 3 to 9 weeks were less well developed in this enzyme than 12-week-old ones.

Fasciola hepatica: mebendazole and thiabendazole pharmacokinetics in sheep.

Pharmacokinetics of the two anthelmintic drugs mebendazole and thiabendazole were determined in sheep before and 4, 8, 13, 19, and 25 weeks after an infestation of animals by an oral administration of 150 metacercariae of Fasciola hepatica. The parasitic pathology was ascertained by the increase in plasma enzyme activities of glutamate dehydrogenase and gamma-glutamyltransferase. After oral administration of mebendazole (25 mg.kg-1), the parent drug and especially its reduced metabolite were present in plasma of animals. A significant 1.5- to 2.7-fold increase in the mean residence time occurred by Weeks 13 to 25 postinfection. This change was related to decreases in both the elimination from the pharmacokinetic compartment representing the reduced metabolite and the area under the curve of plasma metabolite concentration versus time. A 59% decrease in MBZ reduction was demonstrated in liver microsomes prepared from 12-week-infected sheep. This reductase activity was characterized by NADPH dependency and a pH peak activity of 6.0 and was competitively inhibited by daunomycin. In sheep receiving a 50 mg.kg-1 oral dose of thiabendazole, fascioliasis provoked only decreased plasma concentrations of the metabolite 5-hydroxythiabendazole by Weeks 4 to 25 postinfection. This change parallels an increase in urinary excretion of free metabolite but this is of minor significance in the general fate of the drug because of the prevalence of excretion as conjugates. In summary, fascioliasis appears to have more of an effect on the pharmacokinetics of mebendazole, a drug intensively metabolized by the liver into a metabolite present at high concentrations in the plasma of animals and humans.

Shortcomings of the French reimbursement system.

PIP: The vast majority of women living in France are enrolled in the national social security scheme. But very often their eligibility arises through the membership of a husband, cohabitant, or parent paying contributions. The insured pays for medical care, and the social security authorities will reimburse 70-80% of the rates set by the authorities themselves. Only a few brands of pills are refunded, mainly the cheapest ones. No reimbursement is available for the so-called third-generation pills, which cost around 50 francs (US$8) per cycle. IUDs cost around 200 francs (US$32) and are reimbursed at around 70 francs (US$12). The diaphragm can be reimbursed but is very rarely prescribed. Condoms are not reimbursed, but they have recently started selling for 1 franc. Increasingly, private gynecologists are charging much more than social security rates for consultations and the insertion of IUDs. Family planning services are also offered in family planning centers. The IPPF member association in France, MFPF, has about 20 medical centers. The family planning centers offer consultations at the social security rate; to girls under 18, they can be offered free of charge. However, up to the age of 20, young people are eligible only through their parents' insurance. The Ministry of Health rate for an abortion is 1000-1400 francs (US$170-230), plus any additional examinations such as blood tests or ultrasound scans; costs are reimbursed on the basis of 80% of this rate. However, there is a shortage of available beds in clinics; for example, in the Paris region 70% of women cannot find beds. MFPF often deals with women who cannot afford to pay, and those who do not have a residence permit are completely excluded from reimbursement for family planning; but special medical assistance has been set up for women not entitled to Social Security or for those unable to pay.^ieng

Evaluation of erythropoietic/hematopoietic reconstitution after BMT by highly fluorescent reticulocyte counts compares favorably with traditional peripheral blood cell counting.

Highly fluorescent reticulocyte (HFR) counts and reticulocyte counts were analyzed daily using a Sysmex R-1000 automated reticulocyte counter after 42 BMT in children (median age 7 years, range 10 month-18.5 years). White blood cell levels were also analyzed daily using either traditional counting methods or, when levels were very low, cytocentrifuge preparations. Twenty-eight patients received autologous and 14 allogeneic BMT. After myeloablative therapy, HFR counts fell to zero and rose to 2% of the reticulocyte total after a median time of 10 days (range 6-23 days) post-autoBMT and 14 days (range 9-29 days) post-alloBMT. The appearance of HFR (at least 2% in two successive counts) preceded the attainment of 20 x 10(9)/l reticulocytes in 23 of 26 autoBMT by a median time of 3 days, and in 13 of 13 alloBMT with a median time of 4 days (p < 0.001). Two per cent HFR preceded the attainment of 0.5 x 10(9)/l neutrophils in the majority of BMT by a median time > 6 days (p < 0.001). The attainment of 0.05 x 10(9)/l monocytes was preceded by a rise in HFR in 12 of 26 autoBMT and in 7 of 13 alloBMT. Median times between 2% HFR and monocyte levels of 0.05 x 10(9)/l were respectively 2 (range 0- > 24) and 3 (range: 0- > 19) days after auto and alloBMT. These results show that the evaluation of erythropoiesis following BMT by means of HFR counting provides an early measure of hematopoietic reconstitution which is as precise and considerably more practical than monitoring the monocyte level.

Complement activation during low-density lipoprotein apheresis.

Complement system activation was investigated in two girls with familial homozygous hypercholesterolemia undergoing two monthly sessions on LA15 or LA40 (Kaneka liposorber). We determined blood levels of C3c and C3a, leukocyte counts, and plasma levels of C3c and C3a in the extracorporeal circulation device at the start of the sessions and 15 and either 60 or 120 min into them. Sequential eluates were collected from LA40 at the end of the sessions (0.5M NaCl, 1M hydroxylamine). Anaphylatoxin C3a increased throughout, especially with LA40. As previously reported, C3a was trapped in the dextran column but was noticeably present in efferent plasma. Besides many proteins, nonnative complement fragments bearing C3a and C3d antigens were detected in almost all the eluates, suggesting possible in situ complement activation. Practically, complement activation induced by the first filter is a risk; long-term side effects may arise from this extracorporeal circulation device.

[In vitro antibacterial effect of a new oral cephalosporin, cefixime. Results of a multicenter study]. / Activité antibactérienne in vitro d'une nouvelle céphalosporine orale, le céfixime. Résultats d'une étude multicentrique.

Minimal inhibitory concentrations (MIC) of cefixime (CXM) were evaluated by agar dilution against 2,469 bacterial strains isolated in 10 hospitals. For Enterobacteriaceae, MIC 50 and 90% micrograms/ml were respectively: (I) naturally non beta lactamase producing species: E. coli and Shigella 0.25-0.5; Salmonella 0.06-0.25; P. mirabilis 0.008-0.032. (II) chromosomal penicillinase producing species: Klebsiella 0.06-2. (III) chromosomal cephalosporinase producing species: E. cloacae and C. freundii 1-greater than 128; S. marcescens 0.25-16; indole + Proteus 0.06-4; P. stuartii 0.032-0.5. Activity of CXM was not modified against plasmid-mediated penicillinase producing strains, but CXM was inactive on cephalosporinase hyperproducing strains and on broad spectrum beta lactamases producing strains. CXM was inactive on P. aeruginosa (MIC 50 and 90%: 64-128) and on A. baumannii (16-128). Haemophilus and Gonococci, regardless of beta-lactamase production status, and Meningococci were very susceptible to CXM (MIC 0.008-0.12). B. catarrhalis was generally inhibited by 0.03 to 0.5. CXM was poorly active on methicillin susceptible Staphylococci (MIC 50 and 90%: 1-64) and inactive on methicillin resistant strains. Enterococci were generally resistant whereas Streptococci and Pneumococci were inhibited by low concentrations: 0.008 to 1. These antibacterial properties place CXM in excellent position among oral cephalosporins.

RU 486: a new method for early abortion.

PIP: At the end of September 1988, Mifepristone or RU-486, was put on the market in France, to be used in association with prostaglandins to terminate pregnancy during the 1st 7 weeks. Mifepristone stops the pregnancy, and the prostaglandins facilitate the expulsion of the egg. At the initial visit the woman is examined and told about Mifepristone. A week later, she returns and takes 3 tablets of RU-486 in the presence of the physician. 48 hours later she returns to hospital and is given an intramuscular injection of prostaglandin. Bleeding starts within 48 hours and is sometimes heavy and protracted. A week later the woman is given a check-up and an interview. Women find the experience extremely stressful, especially since they themselves carry out the act that terminates the pregnancy.^ieng

Clinical isolates of Staphylococcus lugdunensis and S. schleiferi: bacteriological characteristics and susceptibility to antimicrobial agents.

The bacteriological characteristics and susceptibility to antimicrobial agents of 108 clinical isolates of Staphylococcus lugdunensis and Staphylococcus schleiferi are described. Fifty out of 108 isolates were considered to be responsible for 16 documented infections, including some severe infections (endocarditis, bacteraemia, osteitis). A number of bacteriological characteristics enabled the identification of these species in the clinical microbiology laboratory: the absence of coagulase and protein A, and the presence of a fibrinogen affinity factor and thermonuclease along with other biochemical characteristics (ornithine and arginine decarboxylases, carbohydrate acidification, novobiocin susceptibility) differentiated these new species from other staphylococci; however, they did not possess virulence markers such as toxins or haemagglutinin, but were haemolytic. In this series, almost all isolates were susceptible to 22 antibiotics and 4 antiseptics representative of the main groups of antimicrobial agents. More information is needed on the ecology and epidemiology of these new opportunistic pathogens.

Susceptibilities to ceftriaxone of streptococcal strains associated with infective endocarditis.

We determined the bactericidal activity of ceftriaxone on 20 streptococci isolated from patients with infective endocarditis and that of penicillin G on 5 strains. The MICs of ceftriaxone were less than or equal to 2 micrograms/ml and the MBCs were low for 5 nontolerant strains (less than or equal to 2 micrograms/ml) and high for 15 tolerant strains (greater than or equal to 16 micrograms/ml). The maximal reduction of the viable bacterial counts after 24 h of exposure to antibiotic was achieved for a concentration of ceftriaxone of 4, 32 and 256 micrograms/ml, respectively for 5, 10 and 19 strains. The activity of penicillin G was similar.

[In vitro activity of roxithromycin compared to 5 other macrolides against staphylococci]. / Activité in vitro de la roxithromycine comparée à celle de cinq autres macrolides sur les staphylocoques.

This study aimed to compare the bacteriostatic activity of roxithromycin (RU) to those of erythromycin (ERY), troleandomycin (TAO), spiramycin (SPI), josamycin (JOS) and midekamycin (MID) against staphylococci strains. 239 strains of hospital origin were analysed: S. aureus (139), coagulase negative staphylococci (CNS) (100). The MIC were determined by the agar dilution method. The modal MIC, the MIC 50 and 90 observed for the both groups of strains are given according to species and antibiotics. This study gives the opportunity to classify the 6 antibiotics in a decreasing order considering their antistaphylococcal activity: RU = ERY, TAO = SPI, JOS = MID. No difference was noticeable between S. aureus and CNS strains.

Susceptibilities of streptococcal strains associated with infective endocarditis to nine antibiotics.

The susceptibilities of 121 streptococcal strains isolated from patients with infective endocarditis to mezlocillin, piperacillin, ceftizoxime, ceftriaxone, ceftazidime, imipenem, ciprofloxacin, ofloxacin and pefloxacin were determined by the agar dilution technique. Viridans streptococci, Streptococcus bovis and Enterococcus faecalis were susceptible to imipenem, mezlocillin and piperacillin. All the strains, except E. faecalis were sensitive to ceftriaxone, ceftizoxime and ceftazidime and resistant to ciprofloxacin, ofloxacin and pefloxacin. E. faecalis strains were moderately resistant to the new quinolons.

[Pathology of the isthmo-interstitial segment of the oviduct, tubal pregnancy and microsurgery]. / Pathologie du segment isthmo-interstitiel de l'oviducte, grossesse tubaire et microchirurgie.

With regard to the retrospective and prospective analysis of two series of patients, a first series concerning 180 patients presenting pathology of the isthmo-interstitial segment of the oviduct, and a second series concerning 149 patients operated on for extrauterine pregnancy, the authors have established the responsibility of the pathology of the proximal segment in a very large number of cases of sterility regarded as inexplicable, and in a certain number of cases of extrauterine pregnancy. Medical treatment with danazol gives remarkable results. In cases of failure of medical treatment, microsurgery involves isthmo-ostial reimplantation.