Antibacterial activity of ofloxacin in urine for 4 days after a single oral dose of 400 mg

Antibacterial activity of ofloxacin in urine after a single oral dose of 400 mg was evaluated in ten healthy female volunteers. Urine was collected over six periods, i.e., 0-6 h, 6-12 h, 12-24 h, 24-48 h, 48-72 h, and 72-96 h postdose. Ofloxacin levels were assayed in all samples using a microbiological method and HPLC. Urinary ofloxacin MICs were determined for five bacterial strains recovered from urine, two E. coli strains of which one was susceptible and the other resistant to nalidixic acid (Nal-A), one Klebsiella pneumoniae resistant to nalidixic acid (Nal-B), one Staphylococcus saprophyticus strain, and one Enterococcus faecalis strain; MICs were 0.06, 0.25, 1, 0.25, and 2 mg/L, respectively. Mean urinary ofloxacin levels by the microbiological method during the six collection periods were 193.3 +/- 30.3, 138.1 +/- 31, 53.2 +/- 7.3, 8.3 +/- 0.8, 1.4 +/- 0.2, and 0.6 +/- 0.1 mg/L, respectively. HPLC provided similar results: 216.7 +/- 31.6, 130.7 +/- 20.5, 56.5 +/- 7.1, 8.3 +/- 0.9, 1.5 +/- 0.3, and 0.5 +/- 0.05 mg/L, respectively. Mean urinary ofloxacin excretion over 96 h was 67.4 +/- 3.6% of the dose by the microbiological method was 72.5 +/- 2.5% of the dose by HPLC. On the first day, bacteriostatic activity of urine against enterobacteria exceeded 32 and was greater than 8192 for the nalidixic acid-susceptible E. coli strain; on the next day, overall values were equal or greater than 8 for the nalidixic acid-resistant E. coli and K. pneumoniae strains. Bacteriostatic activity was equal to or greater than 32 for the S. saprophyticus strain during the first two days and equal to 8 on the first day and 4 on the second day for the E. faecalis strain.     

Peyer's patch adherence of enteropathogenic Escherichia coli strains in rabbits

RDEC-1 (serotype O15) is an attaching and effacing strain of rabbit enteropathogenic Escherichia coli (REPEC) that causes diarrhea in postweanling rabbits. It expresses AF/R1 pili that mediate Peyer's patch M-cell adherence. We investigated Peyer's patch adherence, the presence of virulence genes, ileal brush border aggregation, and pilus expression in 9 strains representing several serotypes of REPEC as well as in two commensal strains. Postweanling rabbits were inoculated with 10(6) organisms and sacrificed at 24 h, and tissues were prepared for examination by light microscopy. Strains B10 and RDEC-1 were also studied at 12 and 72 h postinoculation. All REPEC strains were eaeA positive, expressed pili, and adhered to ileal brush borders. Both commensal strains expressed pili, and one strain adhered to brush borders. All REPEC strains demonstrated some degree of Peyer's patch lymphoid follicle adherence, ranging from diffuse coverage to small patches covering two to three dome epithelial cells. Strains C102 and C110 had genes homologous with the structural subunit gene of the AF/R1 pilus (afrA) of RDEC-1, which correlated with greater degrees of lymphoid follicle adherence and lesser degrees of ileal villus adherence. The observation that all REPEC strains adhere to Peyer's patch epithelium suggests the possibility that human strains of enteropathogenic E. coli (EPEC) might do likewise. EPEC strains might thus serve as mucosal vaccine vectors in humans. Better understanding of the molecular mechanism of REPEC adherence should provide a model for the targeting of the Peyer's patch in humans.     

Glycosphingolipids of skeletal muscle: I. Subcellular distribution of neutral glycosphingolipids and gangliosides in rabbit skeletal muscle

The development of polymers with different surface properties and surface modifications of intraocular lenses (IOL) should reduce foreign body reactions after implantation by reducing the surface hydrophobicity of the lenses. It was examined how far such surface variations influenced the adhesiveness of bacteria. The most common organism isolated from cases of postoperative endophthalmitis is Staphylococcus epidermidis. For this reason, three strains of this species, the type strain ATCC 14990 and two clinical isolates (8687, 6579 I), with different hydrophobic surfaces, were studied. IOL made of PMMA, silicone, and a copolymer as well as PMMA lenses with modified surfaces (unpolished, polished, silanized, and heparinized) were used. Bacteria were radiolabelled with 3H-thymidine and the adherent bacteria were calculated per mm2 of lens surface. The three strains adhered better to the unpolished surface of silicone than to PMMA. Treatment of PMMA surface by polishing diminished the differences between the strains. An influence of hydrophobic interactions on the adherence of S. epidermidis ATCC 14990 was demonstrated. The adherence of this hydrophobic type strain was clearly reduced by heparinization of the PMMA surface. In contrast, the hydrophilic catheter isolate 6579 I adhered better to modified surfaces. This strain differed clearly in its PFGE pattern from both hydrophobic strains. Hydrophobic interactions play a role in the bacterial adherence to intraocular lenses in vitro and in vivo. Modifications of polymer surfaces, however, can result in rather different effects depending on the bacterial surface composition and properties.     

Bacterial growth in human vitreous humor

The aim of this study was to investigate the capacity of human vitreous to support bacterial growth and to show differences in the growth kinetics of gram-positive and gram-negative bacteria. Vitreous gel of 70 keratoplasty donor eyes was sampled under sterile conditions, screened microscopically for cellular components and tested for sterility and levels of antibiotic drugs by bio-assay. The samples were inoculated with clinical isolates of Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus viridans and Streptococcus pyogenes. As control each strain was added both to 0.9% sodium chloride solution and to Mueller-Hinton broth. In order to determine bacterial growth the number of colony forming units was determined 4, 6, 24, 48 and 72 hr after inoculation by viable count. Vitreous gel did not support bacterial growth; the tested strains could not be recovered after 48 hr. Similar results could be obtained with sodium chloride; whereas in Mueller Hinton broth the strains showed normal pattern of growth. It seems that vitreous humor has inherent antibacterial capacity in vitro, although the responsible factors remain unknown.     

The effect of hip stem material modulus on surface strain in human femora

Human femora were used to compare the changes in bone surface strain resulting from decreasing the material modulus of a collarless hip stem to determine whether a highly elastic stem increased bone loading. Three substrate materials were tested: titanium (modulus of elasticity 110 GPa), carbon fiber composite (modulus of elasticity 52 GPa), and polymethylmethacrylate (PMMA, modulus of elasticity of 1.9 GPa). Two separate analyses were performed in which femora were implanted randomly with one of the three stem types. Results showed that assembly strains did not differ significantly among different materials. There was a large strain reduction in the proximal region of the femora for all stem substrates relative to the intact femur. Although there was statistically greater surface shear strain as the material modulus decreased, the PMMA stem did not substantially increase bone loading.     

The abilities of a Staphylococcus epidermidis wild-type strain and its slime-negative mutant to induce endocarditis in rabbits are comparable

The abilities of a parent and mutant pair of Staphylococcus epidermidis strains, the slime-producing parent RP62A and its slime-negative mutant, to establish endocarditis in a rabbit model of aortic valve endocarditis and to accumulate and adhere to surfaces in vitro were compared. Vegetation titer and infection rate depended on the presence or absence of a catheter (P = 0.020) and on inoculum size (P < 0.001) but not on the infecting strain. The ability of the parent strain vis-à-vis its mutant to accumulate in vitro on surfaces as demonstrated in a slime test did not correlate with any enhancement in the development of endocarditis in the rabbit model. In vitro initial adherence rates were identical. Both isolates accumulated to the same reduced extent in vitro in the presence of serum, albumin, or gelatin. Adhesion was equally promoted by addition of fibronectin. These data suggest that the in vitro phenomenon of accumulation described as slime production in the absence of serum may not be an important virulence determinant in vivo.     

Antibody and cytokine responses in a mouse pulmonary model of Shigella flexneri serotype 2a infection

A murine pulmonary model was used to study the mucosal immune response to Shigella flexneri serotype 2a infection. Inoculation of BALB/cJ mice with shigellae via the intranasal route resulted in bacterial invasion of bronchial and alveolar epithelia with concomitant development of acute suppurative bronchiolitis and subsequent development of lethal pneumonia. The pathology of pulmonary lesions resembled the colitis that characterizes shigellosis in humans and primates. Significant protection against a lethal dose of S. flexneri 2a was observed in mice previously infected with two sublethal doses of the homologous strain. Immunity against lethal challenge was associated with decreased bacterial invasion of the mucosal epithelium. Over the course of two sublethal challenges, which constituted primary and secondary immunizations, mice developed pulmonary and serum immunoglobulin G and A antibody recognizing both lipopolysaccharide and invasion plasmid antigens IpaB and IpaC. Immune mice and naive control mice differed in lung lavage cytokine levels following lethal challenge. Immune mice developed significantly elevated levels of pulmonary gamma interferon within 6 h of challenge, while naive control mice developed elevated levels of this cytokine later during the initial 24-h period. Both groups had elevated levels of gamma interferon during the 24- to 48-h period of infection. Both groups also had elevated levels of tumor necrosis factor alpha within 6 h of challenge, but the control mice had significantly higher levels at the 48- and 72-h time points. Elevated levels of interleukin-4 were observed only in immunized mice. This cytokine appeared within 24 h and receded between 48 and 72 h. Fluorescence-activated cell sorter analysis of lung parenchymal cells showed that both groups experienced an initial influx of monocytes, but the proportion of this cell type began to recede in immunized mice after 48 h of infection, while peak levels were maintained in the control animals. These studies suggest that elements of local B lymphocyte activity, as well as Th1 and Th2 lymphocyte activity, may contribute to the survival of immune mice after intranasal challenge with shigellae.     

Adherence of staphylococci of different hydrophobicity. Study of various intraocular lenses

BACKGROUND: A major goal in research on intraocular lenses (IOL) is the development of new polymers and modifications to reduce foreign-body reactions after implantation. This effect may be achieved by a reduction in the surface hydrophobicity of the polymers. To illustrate the influence of surface modifications on bacterial adhesiveness, the most often isolated organism in "low-grade" postoperative endophthalmitis, Staphylococcus epidermidis, was used. MATERIALS AND METHODS: For this reason three strains of this species, the type strain ATCC 14990 and two clinical isolates (8687, 6579 I) with different hydrophobic surface properties were studied. IOL, used in the experiments were either made of PMMA or silicone with modified surfaces (unpolished, polished, heparinized). The adhesiveness of H3-thymidin-labeled bacteria was calculated/mm2 of lens surface. Each experiment was performed in triplicate and repeated three times. RESULTS: The hydrophobic-type strain showed stronger adherence to unpolished PMMA surface (8000 bacteria per mm2) compared to the polished (5200 bacteria/mm2). In contrast, the hydrophilic strain adhered with 2000 bacteria/mm2 to the unpolished and with 4200 bacteria/mm2 to the polished surface. Polishing PMMA lenses diminished the differences between the three strains. However, surface passivation of silicone lenses increased the adhesion rate of the hydrophilic strain up to 9600 bacteria/mm2. Treatment of PMMA lenses with heparin increased the adhesiveness of the hydrophilic strain and reduced the adhesion rate of the hydrophobic type strain to 250 bacteria/mm2. CONCLUSIONS: It was demonstrated that bacterial adherence to IOL also involves hydrophobic interactions. Obviously, however, that adherence reflects a complex of interactions between the two surfaces.     

Restricted usage of T cell receptor V alpha/J alpha gene segments with different nucleotide but identical amino acid sequences in HLA-DR3+ sarcoidosis patients

Alkylation of DNA was studied after treatment with [3H]melphalan (phenylalanine mustard; 1-2 microM) using a human tumour cell line, RD, in culture, or Escherichia coli (WP2 or WP2-uvrA strains) in growth medium. After 6 h at 37 degrees C, treated cells were isolated and re-suspended in fresh growth media. Samples were taken at times up to 48 h for isolation of DNA, and in some cases also RNA and protein (which were found to be alkylated to about the same extent as DNA). Alkylated DNA was analysed as previously described (M.R. Osborne and P.D. Lawley, Chem.-Biol. Interact 89 (1993) 49-60). The four principal products, mono-7-alkylguanine (G-M-OH); mono-3-alkyladenine (A-M-OH); and the cross-linked products G-M-G and A-M-G, were identified in DNA from melphalan treated cells, and quantitatively determined. In each case, alkylation of cellular macromolecules was maximal after about 6 h. In DNA of the human tumour cell line, the relative amounts of adenine products decreased with time, most markedly with A-M-OH to 42% of the 2-h value after 48 h. In DNA of both bacterial strains, A-M-OH was virtually undetectable even at early times. Comparisons between the time course of relative decreases in amounts of these alkylpurine products and the corresponding values for alkylated DNA in vitro suggest that the adenine products are subject to removal by repair enzyme action in E. coli of either strain.     

Effect of cultivation medium on some physicochemical parameters of outer bacterial membrane

The changes of surface charge and hydrophobicity of the outer bacterial membrane in relation to utilization of n-hexadecane were studied. For this spectrophotometric study adsorption of methylene blue and transport of gentian violet were used. The decrease in the negative charge of the bacterial strains Pseudomonas putida CCM 3423, P. aeruginosa, and P. fluorescens CCM 2115, depended on the type of growth medium. The decrease of surface charge was in the order: meat extract peptone broth > mineral medium with glucose > mineral medium with n-hexadecane. The highest permeability of the bacterial membrane for gentian violet was determined in the case of P. fluorescens grown in meat extract peptone broth. This effect can be explained by a greater hydrophobicity of the bacterial surface for this strain. In other strains a lower permeability was observed. P. fluorescens showed a greater adherence to hexadecane.     

A highly adherent phenotype associated with virulent Bvg+-phase swine isolates of Bordetella bronchiseptica grown under modulating conditions

The ability of Bvg(-)-phase and Bvg(+)-phase Bordetella bronchiseptica swine isolates, grown under modulating or nonmodulating conditions, to adhere to swine ciliated nasal epithelial cells was determined. When virulent strains were cultivated at 37 degrees C in the Bvg+ phase, numerous adherent bacteria (approximately eight per cell, depending on the strain used) were observed. However, when such strains were grown under modulating conditions (23 degrees C), a significant increase in the level of attachment was seen, suggesting that B. bronchiseptica produces a Bvg-repressed adhesin under these conditions. bvg mutant strains, including an isogenic bvgS mutant, adhered minimally. Western blots indicated that two putative B. bronchiseptica adhesins, filamentous hemagglutinin and pertactin, were not detectable in cultures displaying the highly adherent phenotype. Several proteins apparent in Western blots obtained by using bacterial extracts enriched in outer membrane proteins derived from B. bronchiseptica grown at 23 degrees C were not present in similar extracts prepared from an isogenic bvgS mutant grown at 23 degrees C or from the parent strain grown at 37 degrees C. Adherence of bacteria cultivated at 23 degrees C was almost completely abolished by pretreatment of organisms at 60 degrees C; adherence was reduced by 57% when bacteria were pretreated with pronase E. Temperature shift experiments revealed that the heightened level of adhesion that occurs following growth at 23 degrees C was maintained for up to 18 h when bacteria were subsequently incubated at 37 degrees C. We propose that a Bvg-repressed adhesin, expressed only by modulated bvg+ strains of B. bronchiseptica, may play a key role in the initial colonization of naturally infected swine.     

Viscoelastic behaviour of acrylic bone cements

Local contact stresses at the bone-cement interface are thought to play an important role in the initiation of component loosening. A reduced-modulus bone cement can lower these local contact stresses. The viscoelastic properties of such a cement raised the question of long term subsidence of the implant system. In this study, the viscoelastic properties of a reduced-modulus bone cement were compared with standard polymethylmethacrylate, PMMA, bone cement using stress relaxation tests. Unconstrained stress relaxation tests were performed at 37 degrees C in an aqueous environment by applying 1%, 2.5%, and 5% strains on bone cement specimens and monitoring the diminishing load for 100 h. The initial rapid stress relaxation occurring over the first hour and the steady state stress relaxation occurring between 15 and 100 h were analyzed. A fast stress diminution occurred in PBMMA specimens indicating that, in a total hip arthroplasty application, PBMMA bone cement would transfer the stress quickly and distribute it over a larger area of endosteal bone surface. Steady state stress relaxation experiments showed a significant difference in 2.5% and 5% stress relaxation values (P < 0.05) between PMMA and PBMMA specimens, but not at the 1% stress values. Length measurements indicated that the viscoelastic PBMMA specimens demonstrated little recovery after 100 h of imposed strain whereas the elastic PMMA specimens showed substantial recovery. This seems to indicate relatively larger subsidence rates in unconstrained PBMMA specimens compared to PMMA specimens. In vivo, the cement is surrounded by endosteal bone at the outer side and by an implant on the inner side. Therefore, constrained creep tests are necessary to obtain the data required for an assessment of in vivo subsidence.     

Interleukin-3 plus interleukin-6 may improve chromosomal analysis of multiple myeloma: cytologic and cytogenetic evidence in thirty-four patients

To better define the role of interleukin-3 (IL-3) and IL-6 in the cytogenetic analysis of multiple myeloma (MM), we performed concomitant chromosome and cytologic studies in 34 patients. In each case, 10-30 x 10(6) bone marrow cells were incubated in two independent cultures consisting of conventional cytogenetic medium with and without IL-3 plus IL-6 added for 72 hours. 1-ml aliquots of each culture were aspirated at 24, 48, and 72 hours and exposed to colcemid for 6 hours. Cytospin preparations were then made and mitotic cells were counted and identified as plasma cells or as nonmalignant cells based on their reactivity with an appropriate anti kappa/lambda serum. Slides for conventional cytogenetic analysis were prepared at 72 hours. A greater than two-fold increase of mitotic plasma cells was observed in cytospin preparations from stimulated cultures versus unstimulated cultures in 15 of 34 cases, whereas a less than 2-fold increase, no variation or no mitosis was recorded in 19 cases. Comparison of the number of mitotic plasma cells in stimulated cultures at 24, 48, and 72 hours showed a decreased mitotic activity at 72 hours. Clonal abnormalities were detected by conventional cytogenetic analysis in 19 of 34 cases (55.8%). Recurrent clonal aberrations involved chromosome 13 (4 cases), chromosomes 1p, and 14q (3 cases); chromosomes 3p, 6q, 7q, and 9q (2 cases). We conclude that IL-3 + IL-6 may increase the number of dividing plasma cells in cytogenetic cultures and that a 2-day culture with these cytokines may facilitate the detection of chromosome abnormalities in MM.     

Subzero nonfreezing preservation in a murine limb replantation model

This study was designed to examine the most effective temperature for hypothermic storage, without freezing, to prolong ischemic tolerance in an amputated murine hindlimb model. We measured freezing points in the calf muscle and the subcutaneous tissue of the foot in the amputated limbs of Fisher 344 strain male inbred rats. The highest freezing point was -1.5 degrees C, which was recorded in the calf muscle. To prevent freezing in any of the tissues in the amputated limb, the temperature for the lowest nonfreezing preservation was defined as -1 degrees C. The amputated limbs were preserved at subzero nonfreezing temperature (-1 degrees C) and at 4 degrees C for 4, 8, 12, 24, 48, and 72 h, and were then transplanted to other inbred rats by microsurgical techniques. We evaluated the vascular patency of the anastomoses by direct observation and performed histological examinations on the seventh day after replantation. Subzero nonfreezing preservation of a limb at -1 degrees C for 72 h was significantly superior to hypothermic preservation at 4 degrees C for 72 h in terms of anastomotic patency rates (P < 0.05). The histology of skeletal muscles preserved at -1 degrees C for 8 h showed greater similarity to the normal situation than the histology of those preserved at 4 degrees C for 8 h. Bone viability with osteoblastic activity was maintained in grafted limbs preserved at -1 degrees C for 72 h, but in the limbs preserved at 4 degrees C for 72 h the bone was not viable, showing no osteoblastic activity. Clinically, the period of ischemia in major limb replantation at normal ambient temperatures is limited to about 6 h. In this study, the maximum ischemia time for replantation of a limb containing muscle tissue was prolong to 8 h at -1 degrees C, but the maximum ischemia time at 4 degrees C could not be prolonged to 8 h. Our results suggest that, in the major replantation of a limb containing muscular tissue, hypothermic preservation at -1 degrees C would be more useful than preservation at 4 degrees C.     

Confocal laser scanning microscopy of peritoneal catheter surfaces

Surface topography of used (in situ > 12 months) and unused CAPD catheters was studied by scanning electronmicroscopy (SEM) and confocal laser scanning microscopy (CLSM). Microbial biofilm was observed on all used catheters. Disruption and removal of the attached biofilm revealed extensive pitting of the catheter surface and scoring within the catheter pores. Similar, though less extensive, surface defects were present in unused catheters. Examination by CLSM, with software specific to the determination of surface topography, showed used catheters to have a surface microrugosity greater than that of unused catheters (p < 0.0005). Adherence studies with radiolabelled Staphylococcus epidermidis demonstrated increased adherence to used than to unused catheters (p < 0.0005) after 48 h. However, when catheters were pre-treated with spent dialysate there was a substantial reduction in bacterial adherence to either catheter and no significant difference in adherence to used and unused catheters. Surface microrugosity of CAPD catheters increases during use but is unlikely to be an important factor in bacterial adherence in vivo.     

Pharmacodynamics of vancomycin alone and in combination with gentamicin at various dosing intervals against methicillin-resistant Staphylococcus aureus-infected fibrin-platelet clots in an in vitro infection model

We compared the pharmacodynamic activities of vancomycin with or without gentamicin in an in vitro infection model with methicilin-resistant Staphylococcus aureus-infected fibrin-platelet clots. Infected fibrin-platelet clots (FPCs) were prepared with human cryoprecipitate, human platelets, thrombin, and the organism (approximately 10[9] CFU of MRSA-494/g) and were suspended with monofilament line in an infection model capable of simulating human pharmacokinetics. Antibiotics were bolused to simulate vancomycin regimens of 2 g every 24 h (q24h), 1 g q12h, 500 mg q6h, and continuous infusion (steady-state concentration of 20 microg/ml) and gentamicin regimens of 1.5 mg/kg of body weight q12h and 5 mg/kg once daily (q.d.). Model experiments were performed in duplicate over 72 h. FPCs were removed from the models in quadruplicate at 0, 8, 24, 32, 48, 72 h, weighed, homogenized, diluted, and plated to determine colony counts. The inoculum density at 72 h was used to compare bactericidal activities between the regimens. All regimens containing vancomycin significantly decreased the bacterial inoculum compared to the growth control (P < 0.001). Vancomycin monotherapy regimens were similar in bacterial kill regardless of dosing frequency. The addition of gentamicin (either q12h or q.d.) significantly improved the bactericidal activity of the vancomycin q6h, q12h, and q24h regimens (P < 0.001). The greatest reduction in bacterial density at 72 h (P < 0.001) and the most rapid rate of kill (time to 99.9% killing) were achieved with the regimen consisting of 2 g of vancomycin q24h plus gentamicin (q.d. or q12h).     

胶质芽孢杆菌HJ07的UV与NTG诱变育种及其对铝土矿浸矿效果

以从河南铝土矿样筛选出的一株胶质芽孢杆菌HJ07为出发菌株,对其进行紫外(UV)与亚硝基胍(NTG)诱变育种及铝土矿浸矿脱硅研究.分别通过紫外线照射120 s与采用质量浓度为600 mg·L-1的亚硝基胍处理,出发菌株HJ07的致死率分别达到89%与90%,正突变率分别达到16.5%与18.7%.从突变菌株中筛选所得的两株菌种UV-2与NTG-5的生长代谢活性与脱硅能力明显比出发菌株高.在铝土矿浸出体系中,UV-2与NTG-5达到生长稳定期的时间比HJ07分别缩短了48 h与24 h,且生长稳定期具有更大的细菌浓度.浸矿12 d后,UV-2与NTG-5菌株浸出液中SiO2的质量浓度分别比HJ07提高了约25.6%与12.5%,且达到浸出终点的时间分别缩短了3 d和2 d.UV-2与NTG-5菌株较出发菌株HJ07具有更强的产酸与产胞外聚合物的能力.被UV-2菌株作用后的铝土矿表面的溶蚀程度更加显著,矿物表面形成了明显的菌胶团.      

Effect of citrate on growth of Lactococcus lactis subsp. lactis in milk

The effect of citrate on the growth of Lactococcus lactis subsp. lactis var. diacetylactis in milk has been investigated. Five strains of Lactococcus lactis subsp. lactis var. diacetylactis were compared to their citrate-negative variants, which lack the plasmid coding for citrate permease. In most cases, acidification kinetics and the final bacterial concentration of pure cultures of parental and variant strains did not differ significantly. Co-cultures of parental and variant strains, however, systematically tended towards the predominance of parental strains. Citrate metabolism is responsible for this change, since the predominance of citrate-positive strains was not observed in the absence of citrate. Continuous culture in milk enabled the difference in growth rates between the parental strain Lactococcus lactis subsp. lactis var. diacetylactis CDI1 and its citrate-negative variant to be quantified by following changes in the populations of the two co-cultured strains. At 26 degrees C, the growth rate of the parental strain was 7% higher than that of its citrate-negative variant. These results show that citrate metabolism slightly stimulates the growth of lactococci in milk.     

The significance of enteroaggregative Escherichia coli in the etiology of hospitalized diarrhoea in Calcutta, India and the demonstration of a new honey-combed pattern of aggregative adherence

Previous studies have identified enteroadherent Escherichia coli that exhibit localized adherence, diffuse adherence and atypical diffuse adherence as diarrhoeagenic agents associated with infantile diarrhoea in Calcutta, India. In this study, a DNA probe specific for enteroaggregative adherence was used to determine the etiological significance of enteroaggregative E. coli in the causation of diarrhoea. From a total of 330 strains of E. coli recovered from 159 cases of acute secretory diarrhoea and 174 cases of invasive diarrhoea, 20 strains hybridized with the probe, whereas of the 25 E. coli strains recovered from 25 healthy controls only 1 strain hybridized with the probe. Of the 21 probe positive strains, 19 adhered to HeLa cells in the typical stacked-brick pattern while 2 strains recovered from 2 cases of secretory diarrhoea adhered to the glass surface in a hitherto undescribed formation which we have termed, based on the appearance, as the honey-comb pattern. The enteroaggregative E. coli strains identified in this study did not produce any conventional enterotoxins and were significantly associated with patients with secretory diarrhoea (10.7%) than with invasive diarrhoea (1.7%). The results of this study indicate that enteroaggregative E. coli play a causal role in acute secretory diarrhoea in this part of the world which lends credence to the involvement of a potent toxin in the pathogenesis of EAggEC mediated infections.     

Pharmacodynamics of RP 59500 (quinupristin-dalfopristin) administered by intermittent versus continuous infusion against Staphylococcus aureus-infected fibrin-platelet clots in an in vitro infection model

We evaluated the bactericidal activity of RP 59500 (quinupristin-dalfopristin) against fibrin-platelet clots (FPC) infected with two clinical isolates of Staphylococcus aureus, one constitutively erythromycin and methicillin resistant (S. aureus AW7) and one erythromycin and methicillin susceptible (S. aureus 1199), in an in vitro pharmacodynamic infection model. RP 59500 was administered by continuous infusion (peak steady-state concentration of 6 microg/ml) or intermittent infusion (simulated regimens of 7.5 mg/kg of body weight every 6 h (q6h) q8h, and q12h. FPCs were infected with S. aureus to achieve an initial bacterial density of 10(9) CFU/g. Model experiments were run in duplicate over 72 h. Two FPCs were removed from each model at 0, 12, 24, 36, 48, and 72 h, and the bacterial densities (in CFU per gram) were determined and compared to those of growth control experiments. Additional samples were also removed from the model over the 72-h period for pharmacokinetic evaluation. All regimens significantly (P < or = 0.01) decreased bacterial densities in the infected FPCs for both isolates compared to growth controls. This occurred even though MBCs were equal to or greater than the RP 59500 concentrations achieved in the models. There were no significant differences found between the dosing frequencies and levels of killing when examining each isolate separately. However, examination of the residual bacterial densities (CFU per gram at 72 h) and visual inspection of the overall killing effect (killing curve plots over 72 h) clearly demonstrated a more favorable bactericidal activity against 1199 than against the AW7 isolate. This was most apparent when the q8h and the q12h AW7 regimens were compared to all 1199 treatment regimens by measuring the 72-h bacterial densities (P < or = 0.01). Killing (99.9%) was not achieved against the AW7 isolate. However, a 99.9% kill was demonstrated for all dosing regimens against the 1199 isolate. The area under the concentration-time curve from 0 to 24 h was found to be significantly correlated with reduction in bacterial density for the AW7 isolate (r = 0.74, P = 0.04). No resistance was detected during any experiment for either isolate. RP 59500 efficacy against constitutively erythromycin- and methicillin-resistant S. aureus may be improved by increasing organism exposure to RP 59500 as a function of dosing frequency.