Direct assessment of possible mutations in the 23S rRNA gene encoding macrolide resistance in Chlamydia trachomatis.

Reports of potential treatment failure have raised particular concerns regarding the efficacy of the single dose azithromycin regimen in the treatment of urogenital and anorectal Chlamydia trachomatis (CT) infections. Several factors have been suggested, including heterotypic resistance. Antimicrobial susceptibility testing in CT requires cell culture with serial dilutions of antibiotics, which is laborious and for which there is no standardized testing methodology. One method to partly overcome these difficulties would be to use a genotypic resistance assay, however most current available assays do still require prior CT culture. In order to facilitate the assessment of genotypic resistance directly from clinical samples, without the need for prior culture, the aim of this study was to develop a CT specific PCR assay for the assessment of resistance associated mutations (RAMs) in the 23S rRNA gene, and to evaluate a sample of clinical cases in which CT PCR's remained positive during follow-up despite azithromycin treatment. Neither the in silico analysis nor the analytical specificity testing demonstrated clinically relevant cross-reactivity with other bacterial species. These results in conjunction with the analytical sensitivity demonstrating consistent CT 23S rRNA gene detection in the range of 10e3 IFU/mL, exemplify the assay's apt performance. Although no known macrolide RAMs were detected in the clinical cases, the described assay allows future culture independent macrolide RAM surveillance in CT, and increases accessibility for other laboratories to engage in screening.

Chlamydia trachomatis intra-bacterial and total plasmid copy number in clinical urogenital samples.

Chlamydia trachomatis (CT) increases its plasmid numbers when stressed, as occurs in clinical trachoma samples. Most CT tests target the plasmid to increase the test sensitivity, but some only target the chromosome. We investigated clinical urogenital samples for total plasmid copy numbers to assess its diagnostic value and intra-bacterial plasmid copy numbers to assess its natural variation. Both plasmid and chromosome copies were quantified using qPCR, and the plasmid:chromosome ratio (PCr) calculated in two cohorts: (1) 383 urogenital samples for the total PCR (tPCr), and (2) 42 vaginal swabs, with one half treated with propium-monoazide (PMA) to prevent the quantification of extracellular DNA and the other half untreated to allow for both tPCr and intra-bacterial PCr (iPCr) quantification. Mann-Whitney U tests compared PCr between samples, in relation to age and gender. Cohort 1: tPCr varied greatly (1-677, median 16). Median tPCr was significantly higher in urines than vaginal swabs (32 vs. 11, p < 0.001). Cohort 2: iPCr was more stable than tPCr (range 0.1-3 vs. 1-11). To conclude, tPCr in urogenital samples was much more variable than previously described. Transport time and temperature influences DNA degradation, impacting chromosomal DNA more than plasmids and urine more than vaginal samples. Data supports a plasmid target in CT screening assays to increase clinical sensitivity.

Gut microbiota composition in relation to the metabolic response to 12-week combined polyphenol supplementation in overweight men and women.

BACKGROUND/OBJECTIVES: The intestinal microbiota may have a profound impact on host metabolism. As evidence suggests that polyphenols affect substrate utilization, the present study aimed to investigate the effects of polyphenol supplementation on intestinal microbiota composition in humans. Furthermore, we examined whether (changes in) gut microbiota composition may determine the metabolic response to polyphenol supplementation. SUBJECTS/METHODS: In this randomized, double-blind, placebo (PLA)-controlled trial, 37 overweight and obese men and women (18 males/19 females, 37.8±1.6 years, body mass index: 29.6±0.5 kg/m2) received either epigallocatechin-3-gallate and resveratrol (EGCG+RES, 282 and 80 mg/day, respectively) or PLA for 12 weeks. Before and after intervention, feces samples were collected to determine microbiota composition. Fat oxidation was assessed by indirect calorimetry during a high-fat mixed meal test (2.6 MJ, 61 energy% fat) and skeletal muscle mitochondrial oxidative capacity by means of ex vivo respirometry on isolated skeletal muscle fibers. Body composition was measured by dual-energy X-ray absorptiometry. RESULTS: Fecal abundance of Bacteroidetes was higher in men as compared with women, whereas other assessed bacterial taxa were comparable. EGCG+RES supplementation significantly decreased Bacteroidetes and tended to reduce Faecalibacterium prausnitzii in men (P=0.05 and P=0.10, respectively) but not in women (P=0.15 and P=0.77, respectively). Strikingly, baseline Bacteroidetes abundance was predictive for the EGCG+RES-induced increase in fat oxidation in men but not in women. Other bacterial genera and species were not affected by EGCG+RES supplementation. CONCLUSIONS: We demonstrated that 12-week EGCG+RES supplementation affected the gut microbiota composition in men but not in women. Baseline microbiota composition determined the increase in fat oxidation after EGCG+RES supplementation in men.

Impact of a supportive care service for cancer outpatients: management and reduction of hospitalizations. Preliminary results of an integrated model of care.

PURPOSE: Supportive care in oncology is a primary need for every oncology department nowadays. In 2012, in our institution, a dedicated supportive care service (SCS) was created in order to deal with any need our on-treatment patients might have (e.g. tumour-related or treatment-related symptoms). We hypothesized that this service had a positive impact on the number of unplanned hospitalizations; to confirm our hypothesis, we decided to review admission data in 2011 and 2012. METHODS: Using our internal software, we compared admission data in 2011 (that is, the year before the dedicated service was created) and 2012 (when such service began, that is April of that year). We also made an evaluation of the costs of these hospitalizations. RESULTS: Despite an increase of the number of patients treated in our day hospital (+6.5 %), the number of unplanned hospital admissions decreased by 3.2 % (from 17.3 to 14.1 %). The number of patients accessing to emergency room went from 66 to 61 % (a reduction of 5 %). The costs of these hospitalizations were reduced by 2.2 %. CONCLUSIONS: The introduction of the dedicated SCS in our oncology department caused a net reduction by 3.2 % of the number of unplanned hospitalizations of on-treatment cancer patients.

Adjuvant chemotherapy seems beneficial for invasive intraductal papillary mucinous neoplasms.

AIMS: The incidence of intraductal papillary mucinous neoplasm (IPMN) is rising and these neoplasms now represent up to 25% of resected pancreatic neoplasms. The optimal postoperative management of resected invasive IPMN is still debated in the absence of large prospective clinical trials and of validated prognostic factors in this setting. The objective of our study was to identify potential prognostic factors and to investigate the role of adjuvant therapies for patients radically resected for invasive IPMN. METHODS: We retrospectively reviewed clinical and pathological data regarding a large series of patients with invasive IPMN who underwent surgical resection in the last six years at University Hospital of Pisa. RESULTS: Sixty-four patients were considered for the analysis, thirty-three of whom received adjuvant chemotherapy with gemcitabine. In our series node involvement and high tumoral grade emerged as the major pathologic prognostic factors. Patients treated with adjuvant chemotherapy with gemcitabine experienced a longer disease-free survival than those who received surgery alone. CONCLUSIONS: Gemcitabine-based chemotherapy seems beneficial as adjuvant treatment for patients with resected invasive IPMN.

Pharmacokinetics of cefpodoxime in plasma and subcutaneous fluid following oral administration of cefpodoxime proxetil in male beagle dogs.

Pharmacokinetics of cefpodoxime in plasma (total concentration) and subcutaneous fluid (free concentration using microdialysis) was investigated in dogs following single oral administration of prodrug cefpodoxime proxetil (equivalent to 5 and 10 mg/kg of cefpodoxime). In a cross over study design, six dogs per dose were utilized after a 1 week washout period. Plasma, microdialysate, and urine samples were collected upto 24 h and analyzed using high performance liquid chromatography. The average maximum concentration (C(max) ) of cefpodoxime in plasma was 13.66 (±6.30) and 27.14 (±4.56) µg/mL with elimination half-life (t(1/2) ) of 3.01 (±0.49) and 4.72 (±1.46) h following 5 and 10 mg/kg dose, respectively. The respective average area under the curve (AUC(0-∞) ) was 82.94 (±30.17) and 107.71 (±30.79) µg·h/mL. Cefpodoxime was readily distributed to skin and average free C(max) in subcutaneous fluid was 1.70 (±0.55) and 3.06 (±0.93) µg/mL at the two doses. Urinary excretion (unchanged cefpodoxime) was the major elimination route. Comparison of subcutaneous fluid concentrations using pharmacokinetic/pharmacodynamic indices of fT(>MIC) indicated that at 10 mg/kg dose; cefpodoxime would yield good therapeutic outcome in skin infections for bacteria with MIC(50) upto 0.5 µg/mL while higher doses (or more frequent dosing) may be needed for bacteria with higher MICs. High urine concentrations suggested cefpodoxime use for urinary infections in dogs.

Interdependence of primary and Johari-Goldstein secondary relaxations in glass-forming systems.

We report evidence from broadband dielectric spectroscopy that the dynamics of the primary alpha- and secondary Johari-Goldstein (JG) beta-processes are strongly correlated in different glass-forming systems over a wide temperature T and pressure P range, in contrast with the widespread opinion of statistical independence of these processes. The alpha-beta mutual dependence is quantitatively confirmed by (a) the overall superposition of spectra measured at different T-P combinations but with an invariant alpha-relaxation time; (b) the contemporary scaling of the isothermal-pressure and isobaric-temperature dependences of the alpha-and beta-relaxation times as plotted versus the reduced variable Tg(P)/T where Tg is the glass transition temperature. These novel and model-independent evidences indicate the relevance of the JG relaxation phenomenon in glass transition, often overlooked by most current theories.

Microbiological assay for gatifloxacin in pharmaceutical formulations.

A simple, sensitive and specific agar diffusion bioassay for the antibacterial gatifloxacin was developed using a strain of Bacillus subtilis ATCC 9372 as the test organism. Gatifloxacin could be measured in tablets and raw material at concentration ranging 4-16 microgml(-1). The calibration graph for gatifloxacin was linear from 4.0 to 16.0 microgml(-1). A prospective validation of the method demonstrated that the method was linear (r2=0.9993), precise (R.S.D.=1.14%) and accurate. The results confirmed its precision and did not differ significantly from others methods described in the literature. The validated method yielded good results in terms of the range, linearity, precision, accuracy, specificity and recovery. We concluded that the microbiological assay is satisfactory for in vitro quantification of the antibacterial activity of gatifloxacin.

Emergence of a new feature in the high pressure-high temperature relaxation spectrum of tri-propylene glycol.

We investigated dielectric relaxation of a tri-propylene glycol system under high compression. By increasing temperature and pressure we observed that a new relaxation process emerges from the low frequency tail of the structural peak. This new peak starts to be visible at about 0.5 GPa and becomes clearly evident at 1.7 GPa. However, this additional peak merges again with the structural one as the glass transition is approached, since it has a weaker temperature dependence. This finding enriches the relaxation scenario of molecular glass formers confirming that the application of very high hydrostatic pressure can favor the detection of new relaxation or otherwise unresolved processes in supercooled liquid systems.

Dynamics of supercooled and glassy dipropyleneglycol dibenzoate as functions of temperature and aging: Interpretation within the coupling model framework.

Dielectric relaxation measurements of a typical small molecular glassformer, dipropyleneglycol dibenzoate show the presence of two secondary relaxations. Their dynamic properties differ in the equilibrium liquid and glassy states, as well as the changes during structural recovery after rapid quenching the liquid to form a glass. These differences enable us to identify the slower secondary relaxation as the genuine Johari-Goldstein (JG) beta-relaxation, acting as the precursor of the primary alpha-relaxation. Agreement between the JG beta-relaxation time and the independent relaxation time of the coupling model leads to predicted quantitative relations between the JG beta-relaxation and the alpha-relaxation that are supported by the experimental data.

Adam-Gibbs model for the supercooled dynamics in the ortho-terphenyl ortho-phenylphenol mixture.

Dielectric measurements of the alpha-relaxation time were carried out on a mixture of ortho-terphenyl (OTP) with ortho-phenylphenol, over a range of temperatures at two pressures, 0.1 and 28.8 MPa. These are the same conditions for which heat capacity, thermal expansivity, and compressibility measurements were reported by Takahara et al. [S. Takahara, M. Ishikawa, O. Yamamuro, and T. Matsuo, J. Phys. Chem. B 103, 3288 (1999)] for the same mixture. From the combined dynamic and thermodynamic data, we determine that density and temperature govern to an equivalent degree the variation of the relaxation times with temperature. Over the measured range, the dependence of the relaxation times on configurational entropy is in accord with the Adam-Gibbs model, and this dependence is invariant to pressure. Consistent with the implied connection between relaxation and thermodynamic properties, the kinetic and thermodynamic fragilities are found to have the same pressure independence. In comparing the relaxation properties of the mixture to those of neat OTP, density effects are stronger in the former, perhaps suggestive of less efficient packing.

Gelatin microparticles containing propolis obtained by spray-drying technique: preparation and characterization.

Gelatin microparticles containing propolis extractive solution (PES) were prepared by spray-drying technique. The optimization of the spray-drying operating conditions and the proportions of gelatin and mannitol were investigated. Regular particle morphology was obtained when mannitol was used, whereas mannitol absence produced a substantial number of coalesced and agglomerated microparticles. Microparticles had a mean diameter of 2.70 microm without mannitol and 2.50 microm with mannitol. The entrapment efficiency for propolis of the microparticles was upto 41% without mannitol and 39% with mannitol. The microencapsulation by spray-drying technique maintained the activity of propolis against Staphylococcus aureus. These gelatin microparticles containing propolis would be useful for developing intermediary or eventual propolis dosage form without the PES' strong and unpleasant taste, aromatic odour, and presence of ethanol.

Relaxation processes in an epoxy resin studied by time-resolved optical Kerr effect.

The dynamics of the epoxy resin phenyl glycidyl ether, a fragile glass-forming liquid, is investigated in the liquid and supercooled phases by time-resolved optical Kerr effect experiment with an heterodyne detection technique. We tested the mode-coupling theory and found that the predicted dynamic scenario allows to reproduce properly the measured signal, for t>1 ps, in the whole temperature interval investigated. Furthermore, the values of T(c) and lambda, obtained from the analysis of three different and independent dynamic regimes (alpha regime, von Schweidler, beta regime), are in remarkable agreement. Moreover, relaxation times obtained from optical Kerr effect and dielectric spectroscopy measurements are compared. The two time scales differ only for a constant factor in the whole temperature interval investigated.

Effect of pressure on the dynamics of glass formers.

A description of the pressure dependence of the structural relaxation time has been derived from the Adam-Gibbs theory by writing the configurational entropy in terms of the excess heat capacity and the molar thermal expansion. This new equation was tested successfully on dielectric relaxation data for an epoxy compound over a wide range of temperature and pressure.

Pressure dependence of structural relaxation time in terms of the Adam-Gibbs model.

A new equation describing the behavior of the structural relaxation time, tau(T,P), as a function of both pressure and temperature, is discussed. This equation has been derived from the Adam-Gibbs theory by writing the configurational entropy, S(c), in terms of the excess thermal heat capacity and of the molar thermal expansion. Consequently, the parameters introduced in the expression are directly related to specific physical properties of the material, such as the thermal expansion coefficient alpha and the isothermal bulk modulus K0. At a fixed pressure, for low pressures, the found equation reduces to a Vogel-Fulcher-Tammann equation of tau versus temperature with the fragility parameter independent from pressure. The equation for tau(T,P) was successfully tested directly by fitting the dielectric relaxation time data for two isothermal and one isobaric measurements on diglycidyl ether of bisphenol-A, carried out in previous experiments. The parameters estimated by the best fit were in reasonable agreement with the values determined from the known physical properties of the material. Finally, the expression for the change versus pressure of the temperatures at which the same value of tau(max) is obtained (e.g., the change versus pressure of the glass transition temperature) agrees with several expressions previously proposed in the literature to provide a phenomenological description of the observed phenomena.

Mucormycosis manifesting as proptosis and unilateral blindness.

A 51-year-old woman presented to the emergency department (ED) of another institution with sudden onset of blindness in the left eye. The patient was found to have no light perception in the left eye and a marked chemosis occurring several days after a fall. She was transferred to the hospital for ophthalmologic evaluation. Upon careful history and physical examination, the diagnosis of rhinocerebral mucormycosis was considered and urgent ophthalmology and otolaryngology consults were obtained. The patient underwent extensive surgical debridement and pharmacologic treatment. The diagnosis was confirmed by pathological specimens. In this case report, the clinical presentation, pathogenesis, diagnostic workup, and ED management of mucormycosis are discussed, highlighting the possible diagnostic and therapeutic pitfalls that are most pertinent to the emergency physician.