Guidelines on fibrinogen assays

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Antimicrobial and antioxidant screening of curcumin and pyrocatechol in the prevention of biodiesel degradation: oxidative stability

Owing to its hygroscopicity biodiesel may accumulate water during storage, which becomes favorable to the growth of microorganisms. In order to control microbial contamination, use of various chemical biocides has been studied. However, the addition of a natural substance simultaneously with antioxidant and microbial growth inhibition could prove advantageous in the prevention of biodiesel oxidation and microbial contamination. Curcumin and pyrocatechol are antioxidant agents, which also exhibit microbial growth inhibition abilities. This research effort aimed at evaluating the addition of curcumin and pyrocatechol to biodiesel produced from various vegetable sources (waste frying oil, soybean oil, cottonseed oil, sesame oil, macaúba almond oil and microalgae oil). The combined addition of 1% (w/w) water and curcumin (viz. 0.2% (w/w) for biodiesel from spent frying oil, 0.5% (w/w) for biodiesel from soybean oil, 0.1% (w/w) for biodiesel from cotton seed oil, 0.5% (w/w) for biodiesel from sesame seed oil, 0.2% (w/w) for biodiesel from macaúba almond oil, and 0.2% (w/w) for biodiesel from microalgae oil) were those processing variables that promoted the best fungistatic and antioxidant effects, allowing maintenance of an unfavorable environment for microbial growth in biodiesel inoculated with the ubiquitous filamentous mold Paecilomyces variotii Bainier.Project funding by CNPq (National Council for Scientific and Technological Development - Brazil) (CNPq Ref. No. 404808/ 2013-1, Project ‘Studies on Biodiesel: Development of analytical methods for the characterization and quality control, and research of new natural additives to improve the quality of this biofuel’), is hereby gratefully acknowledged. This work received support from CNPq, National Council for Scientific and Technological Development Brazil, in the form of Research Productivity (PQ) fellowships granted to Victor M. Balcão (Ref. No. 306113/2014-7) and Marco V. Chaud (Ref. No. 309598/2014-1). The authors have no conflicts of interest whatsoever to declare

Evaluation of the best method to assess antibiotic potentiation by phytochemicals against staphylococcus aureus

The increasing occurrence of bacterial resistance to antibiotics has now reached a critical level. Finding antibiotic coadjuvants capable to inhibit the bacterial resistance mechanisms would be a valuable mid-term solution, until new classes of antibiotics are discovered. Selected plant alkaloids were combined with 5 antibiotics against 10 Staphylococcus aureus strains, including strains expressing distinct efflux pumps and methicillin-resistant S. aureus strains. The efficacy of each combination was assessed using the microdilution checkerboard, time-kill, Etest, and disc diffusion methods. The cytotoxicity of the alkaloids was evaluated in a mouse fibroblast cell line. Potentiation was obtained in 6% of all 190 combinations, especially with the combination of: ciprofloxacin with reserpine (RES), pyrrolidine (PYR), and quinine (QUIN); tetracycline with RES; and erythromycin with PYR. The highest cytotoxicity values were found for QUIN (half maximal inhibitory concentration [IC50] = 25 ± 2.2 mg/L) and theophylline (IC50 = 100 ± 4.7 mg/L).The authors are very grateful to Professor Simon Gibbons (Department of Pharmaceutical and Biological Chemistry, The School of Pharmacy, UCL School of Pharmacy, London) for providing some of the bacterial strains. This work was supported by Operational Programme for Competitiveness Factors - COMPETE and by FCT Portuguese Foundation for Science and Technology through Projects Bioresist - PTDC/EBB-EBI/105085/2008; Phytodisinfectants - PTDC/DTP-SAP/1078/2012 and the PhD grants awarded to Ana Abreu (SFRH/BD/84393/2012) and Anabela Borges (SFRH/BD/63398/2009)

Suboptimal clinical response to ciprofloxacin in patients with enteric fever due to Salmonella spp. with reduced fluoroquinolone susceptibility: a case series

BACKGROUND: Salmonella spp. with reduced susceptibility to fluoroquinolones have higher than usual MICs to these agents but are still considered "susceptible" by NCCLS criteria. Delayed treatment response to fluoroquinolones has been noted, especially in cases of enteric fever due to such strains. We reviewed the ciprofloxacin susceptibility and clinical outcome of our recent enteric fever cases. METHODS: Salmonella enterica Serotype Typhi (S. Typhi) and Serotype Paratyphi (S. Paratyphi) blood culture isolates (1998–2002) were tested against nalidixic acid by disk diffusion (DD) and agar dilution (AD) and to ciprofloxacin by AD using NCCLS methods and interpretive criteria. Reduced fluoroquinolone susceptibility was defined as a ciprofloxacin MIC of 0.125–1.0 mg/L. The clinical records of patients treated with ciprofloxacin for isolates with reduced fluoroquinolone susceptibility were reviewed. RESULTS: Seven of 21 (33%) S. Typhi and S. Paratyphi isolates had reduced susceptibility to fluoroquinolones (MIC range 0.125–0.5 mg/L). All 7 were nalidixic acid resistant by DD (no zone) and by AD (MIC 128- >512 mg/L). The other 14 isolates were nalidixic acid susceptible and fully susceptible to ciprofloxacin (MIC range 0.015–0.03 mg/L). Five of the 7 cases were treated initially with oral ciprofloxacin. One patient remained febrile on IV ciprofloxacin until cefotaxime was added, with fever recurrence when cefotaxime was discontinued. Two continued on oral or IV ciprofloxacin alone but had prolonged fevers of 9–10 days duration, one was switched to IV beta-lactam therapy after remaining febrile for 3 days on oral/IV ciprofloxacin and one was treated successfully with oral ciprofloxacin. Four of the 5 required hospitalization. CONCLUSIONS: Our cases provide further evidence that reduced fluoroquinolone susceptibility of S. Typhi and S. Paratyphi is clinically significant. Laboratories should test extra-intestinal Salmonella spp. for reduced fluoroquinolone susceptibility

Validation of serum IGF-I as a biomarker to monitor the bioactivity of exogenous growth hormone agonists and antagonists in rabbits

The development of new growth hormone (GH) agonists and growth hormone antagonists (GHAs) requires animal models for pre-clinical testing. Ideally, the effects of treatment are monitored using the same pharmacodynamic marker that is later used in clinical practice. However, intact rodents are of limited value for this purpose because serum IGF-I, the most sensitive pharmacodynamic marker for the action of GH in humans, shows no response to treatment with recombinant human GH and there is little evidence for the effects of GHAs, except when administered at very high doses or when overexpressed. As an alternative, more suitable model, we explored pharmacodynamic markers of GH action in intact rabbits. We performed the first validation of an IGF-I assay for the analysis of rabbit serum and tested precision, sensitivity, linearity and recovery using an automated human IGF-I assay (IDS-iSYS). Furthermore, IGF-I was measured in rabbits of different strains, age groups and sexes, and we monitored IGF-I response to treatment with recombinant human GH or the GHA Pegvisomant. For a subset of samples, we used LC-MS/MS to measure IGF-I, and quantitative western ligand blot to analyze IGF-binding proteins (IGFBPs). Although recovery of recombinant rabbit IGF-I was only 50% in the human IGF-I assay, our results show that the sensitivity, precision (1.7–3.3% coefficient of variation) and linearity (90.4–105.6%) were excellent in rabbit samples. As expected, sex, age and genetic background were major determinants of IGF-I concentration in rabbits. IGF-I and IGFBP-2 levels increased after single and multiple injections of recombinant human GH (IGF-I: 286±22 versus 434±26 ng/ml; P<0.01) and were highly correlated (P<0.0001). Treatment with the GHA lowered IGF-I levels from the fourth injection onwards (P<0.01). In summary, we demonstrated that the IDS-iSYS IGF-I immunoassay can be used in rabbits. Similar to rodents, rabbits display variations in IGF-I depending on sex, age and genetic background. Unlike in rodents, the IGF-I response to treatment with recombinant human GH or a GHA closely mimics the pharmacodynamics seen in humans, suggesting that rabbits are a suitable new model to test human GH agonists and antagonists

The role of polysaccharide intercellular adhesin (PIA) in Staphylococcus epidermidis adhesion to host tissues and subsequent antibiotic tolerance

The aim of this study was to determine the role of polysaccharide intercellular adhesin (PIA) in Staphylococcus epidermidis adhesion to host tissues and subsequent antibiotic tolerance. The adherence of S. epidermidis 1457 and the mutant defective in PIA production (1457-M10) to urinary epithelium and endothelium was estimated by colony counting. Minimum bactericidal concentration and mean reduction of cellular activity (XTT) following antibiotic exposure was determined for planktonic and adhered bacteria. S. epidermidis 1457 adhered to a greater extent to both cells than the mutant strain. The adhered strains had a significantly higher antimicrobial tolerance than their planktonic counterparts. The mutant strain was, in general, the most susceptible to the antibiotics assayed. In conclusion, PIA may influence S. epidermidis adherence to host tissues and their antimicrobial susceptibility. Initial adhesion may be the main step for the acquisition of resistance in S. epidermidis

Fungaemia due to Cryptococcus laurentii and a review of non- neoformans cryptococcaemia

Cryptococcus laurentii is one of several non- neoformans cryptococci that have rarely been associated with human infection. The spectrum of clinical infection due to non- neoformans species ranges from skin lesions to fungaemia. Most cases of non- neoformans fungaemia have been noso-comially acquired and have been associated with indwelling intravascular catheters and neutropenia. Limited data on in vitro susceptibilities of non- neoformans cryptococci show these species to be more resistant to fluconazole and flucytosine than most Cr. neoformans. Two such cases are presented here. Zusammenfassung . Cryptococcus laurentii ist eine von mehreren nicht- neofomans-Cryptococcus -Arten, die selten mit Krankheiten am Menschen in Zusammenhang gebracht werden. Das Spektrum durch Nicht- neoformans -Arten bedingter klinischer Infektionen reicht von HautlÄsionen bis zur FungÄmie. Die Mehrzahl der FÄlle von nicht- neoformans -FungÄmie waren nosokomial bedingt und mit intravasalen Dauerkathetern und Neutropenie assoziiert. Die wenigen Daten Über In-vitro-SuszeptibilitÄt von Non- neoformans -Arten fÜr Fluconazol und Flucytosin sprechen dafÜr, daß diese Arten resistenter als die meisten Cr. neoformans-StÄmme sind. Zwei solcher FÄlle werden vorgestellt.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72398/1/j.1439-0507.1998.tb00338.x.pd

Frequency of antimicrobial resistance among invasive and colonizing Group B Streptococcal isolates

BACKGROUND: Group B Streptococcus (GBS) remains susceptible to penicillin, however, resistance to second-line antimicrobials, clindamycin and erythromycin, has increased since 1996. We describe the age-specific antibiotic susceptibility profile and capsular type distribution among invasive and colonizing GBS strains. METHODS: We tested 486 invasive GBS isolates from individuals of all ages collected by a Wisconsin surveillance system between 1998 and 2002 and 167 colonizing strains collected from nonpregnant college students during 2001 in Michigan. Antimicrobial susceptibility testing was performed by disk diffusion or Etest and capsular typing was performed using DNA dot blot hybridization RESULTS: 20.0% (97/486) of invasive and 40.7% (68/167) of colonizing isolates were resistant to clindamycin (P < .001) and 24.5% (119/486) of invasive and 41.9% (70/167) of colonizing isolates were resistant to erythromycin (P < .001). Similarly, 19.8% (96/486) of invasive and 38.3% (64/167) of colonizing isolates were resistant to both antimicrobial agents (P < .001). 29.4% (5/17) of invasive isolates from persons 18–29 years of age and 24.3% (17/70) of invasive isolates from persons 30–49 years of age were resistant to clindamycin. Similarly, 35.3% (6/17) of invasive isolates from persons 18–29 years of age and 31.4% (22/70) of invasive isolates from persons 30–49 years of age were resistant to erythromycin. 34.7% (26/75) of invasive isolates from persons < 1 year of age were capsular type Ia, whereas capsular type V predominated among isolates from adults. CONCLUSION: Clindamycin and erythromycin resistance rates were high among isolates colonizing nonpregnant college students and invasive GBS isolates, particularly among the colonizing isolates. Susceptibility profiles were similar by age although the proportion of clindamycin and erythromycin resistance among invasive isolates was highest among persons 18–49 years of age. Increasing antimicrobial resistance has implications for GBS disease treatment and intrapartum prophylaxis among penicillin intolerant patients

Three Cases of Moraxella osloensis Meningitis: A Difficult Experience in Species Identification and Determination of Clinical Significance

We had three cases of Moraxella osloensis meningitis. The species identification was impossible by conventional and commercial phenotypic tests. However, we could identify the species using the 16S rRNA gene sequencing. Determination of clinical significance was difficult in one patient. All three patients recovered by appropriate antimicrobial therapy