Nonthermal Plasma Jet Treatment Negatively Affects the Viability and Structure of Candida albicans SC5314 Biofilms.

Microorganisms are predominantly organized in biofilms, where cells live in dense communities and are more resistant to external stresses than are their planktonic counterparts. With in vitro experiments, the susceptibility of Candida albicans biofilms to a nonthermal plasma treatment (plasma source, kINPen09) in terms of growth, survival, and cell viability was investigated. C. albicans strain SC5314 (ATCC MYA-2876) was plasma treated for different time periods (30 s, 60 s, 120 s, 180 s, 300 s). The results of the experiments, encompassing CFU, fluorescence Live/Dead, and 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt (XTT) assays, revealed a negative influence of the plasma treatment on the proliferation ability, vitality, and metabolism of C. albicans biofilms, respectively. Morphological analysis of plasma-treated biofilms using atomic force microscopy supported the indications for lethal plasma effects concomitant with membrane disruptions and the loss of intracellular fluid. Yielding controversial results compared to those of other publications, fluorescence and confocal laser scanning microscopic inspection of plasma-treated biofilms indicated that an inactivation of cells appeared mainly on the bottom of the biofilms. If this inactivation leads to a detachment of the biofilms from the overgrown surface, it might offer completely new approaches in the plasma treatment of biofilms. Because of plasma's biochemical-mechanical mode of action, resistance of microbial cells against plasma is unknown at this state of research.IMPORTANCE Microbial communities are an increasing problem in medicine but also in industry. Thus, an efficient and rapid removal of biofilms is becoming increasingly important. With the aid of the kINPen09, a radiofrequency plasma jet (RFPJ) instrument, decisive new findings on the effects of plasma on C. albicans biofilms were obtained. This work showed that the inactivation of biofilms takes place mainly on the bottom, which in turn offers new possibilities for the removal of biofilms by other strategies, e.g., mechanical treatment. This result demonstrated that nonthermal atmospheric pressure plasma is well suited for biofilm decontamination.

Global Burden of Sugar-Related Dental Diseases in 168 Countries and Corresponding Health Care Costs.

Oral diseases such as dental caries, edentulism (tooth loss), periodontal disease (PD), and oral cancer currently constitute an increased major public health burden across the globe, with significant differences between countries. One of the main drivers of caries, edentulism, and PD is the excessive intake of sugars. Here, we aimed to quantify the global sugar-related dental health and cost burden in the year 2010. This study used a health-econometrical model to calculate the disease burden as well as the direct and indirect costs attributable to the intake of free sugars (mono- and disaccharides [MDS]). To this end, several databases from the Institute for Health Metrics and Evaluation (IHME), Organisation for Economic Co-operation and Development (OECD), Food and Agriculture Organization (FAO), and World Bank were used. In total, the corresponding disease burden in 168 countries and economic burden in 31 OECD countries were quantified. In 2010, the consumption of MDS was associated with a global dental disease burden of 4.1 million disability-adjusted life years (DALYs; 95% uncertainty interval [UI]: 2.1 to 7.4 million DALYs), with 2.7 million DALYs from MDS-related caries and 1.4 million DALYs from PD. In terms of economic costs, MDS-related dental diseases were associated with a global financial burden of 172 billion US dollars (USD; 95% UI: 91 to 295 billion USD), the largest share of which (151 billion USD) was incurred in OECD countries. Overall, 26.3% (95% UI: 13.3% to 47.5%) of the total global oral disease burden was attributed to the consumption of MDS. The present study emphasizes the need to further address the role of free sugars in oral health and nutrition policy. Although the largest share of the economic burden was accounted for by OECD countries, emerging economies should address this challenge early on in national public health policies if they are to avoid disease and the prospect of increased cost burdens.

Human cell-based taste perception - a bittersweet job for industry.

Covering: 2000 to 2016On the molecular level humans sense food by a variety of specialized tissues which express sensory receptors to handle nutritive value. In general, this means the interplay of gustatory, olfactory, trigeminal and haptic sensation is translated into perception and leads, in terms of taste, to descriptions like sweet, bitter, salty, sour and umami. Further perceptions include astringent, cool, hot, prickle, lingering, kokumi and fatty to name predominant characterizations. It is still not fully understood how this plethora of impressions can be perceived by quite a limited number of receptors obviously being the initial compilers to judge palatability. However, since the discovery of mammalian taste receptors (TASRs) almost 30 years ago the use of taste receptors in cell-based screening campaigns is advancing in industrial approaches. The article will highlight the impacts and the limits of cell-based guided identification of taste modulators for food applications with an emphasis on sweet, bitter and savory taste as well as implications emerging from natural products.

Ten years of subproteome investigations in lactic acid bacteria: A key for food starter and probiotic typing.

The definition of safety and efficacy of food-employed bacteria as well as probiotic strains is a continuous, often unattended, challenge. Proteomic techniques such as 2DE, DIGE and LC/LC-MS/MS are suitable and powerful tools to reveal new aspects (positive and negative) of "known" and "unknown" strains that can be employed in food making and as nutraceutical supplements for human health. Unfortunately, these techniques are not used as extensively as it should be wise. The present report describes the most significant results obtained by our research group in 10years of study on subproteomes in bacteria, chiefly lactic acid bacteria. Production of desired and undesired metabolites, differences between strains belonging to same species but isolated from different ecological niches, the effect of cryoprotectants on survival to lyophilization as well as the adhesive capability of strains, were elucidated by analysis of cytosolic, membrane-enriched, surface and extracellular proteomes. The present review opens a window on a yet largely underexplored field and highlights the huge potential of subproteome investigations for more rational choice of microbial strains as food starters, probiotics and for production of nutraceuticals. These analyses will hopefully contribute to manufacturing safer and healthier food and food supplements in the near future. This article is part of a Special Issue entitled: HUPO 2014.

Pharmacokinetic and pharmacogenomic modelling of the CYP3A activity marker 4ß-hydroxycholesterol during efavirenz treatment and efavirenz/rifampicin co-treatment.

OBJECTIVES: To assess the effect of the major efavirenz metabolizing enzyme (CYP2B6) genotype and the effects of rifampicin co-treatment on induction of CYP3A by efavirenz. PATIENTS AND METHODS: Two study arms (arm 1, n = 41 and arm 2, n = 21) were recruited into this study. In arm 1, cholesterol and 4ß-hydroxycholesterol were measured in HIV treatment-naive patients at baseline and then at 4 and 16 weeks after initiation of efavirenz-based antiretroviral therapy. In arm 2, cholesterol and 4ß-hydroxycholesterol were measured among patients taking efavirenz during rifampicin-based tuberculosis (TB) treatment (efavirenz/rifampicin) just before completion of TB treatment and then serially following completion of TB treatment (efavirenz alone). Non-linear mixed-effect modelling was performed. RESULTS: A one-compartment, enzyme turnover model described 4ß-hydroxycholesterol kinetics adequately. Efavirenz treatment in arm 1 resulted in 1.74 (relative standard error = 15%), 3.3 (relative standard error = 33.1%) and 4.0 (relative standard error = 37.1%) average fold induction of CYP3A for extensive (CYP2B6*1/*1), intermediate (CYP2B6*1/*6) and slow (CYP2B6*6/*6) efavirenz metabolizers, respectively. The rate constant of 4ß-hydroxycholesterol formation [mean (95% CI)] just before completion of TB treatment [efavirenz/rifampicin co-treatment, 7.40 × 10(-7) h(-1) (5.5 × 10(-7)-1.0 × 10(-6))] was significantly higher than that calculated 8 weeks after completion [efavirenz alone, 4.50 × 10(-7) h(-1) (4.40 × 10(-7)-4.52 × 10(-7))]. The CYP3A induction dropped to 62% of its maximum by week 8 of completion. CONCLUSIONS: Our results indicate that efavirenz induction of CYP3A is influenced by CYP2B6 genetic polymorphisms and that efavirenz/rifampicin co-treatment results in higher induction than efavirenz alone.

Pharmacogenetic and pharmacokinetic aspects of CYP3A induction by efavirenz in HIV patients.

We investigated the effects of pharmacogenetic variations and efavirenz pharmacokinetics on inter-individual differences in the extent of CYP3A induction by efavirenz using 4ß-hydroxycholesterol/cholesterol (4ß-OHC/Chol) as a marker for CYP3A induction. Plasma 4ß-hydroxycholesterol and cholesterol concentrations were determined at baseline, and at the 4th, 16th and 48th week of efavirenz-based highly active antiretroviral therapy in antiretroviral therapy-naive HIV patients (n=77). Efavirenz plasma concentrations were quantified at weeks 4 and 16. CYP2B6, CYP3A5, ABCB1, UGT2B7 genotyping were done. Compared with baseline, the median plasma 4ß-OHC/Chol ratio increased at the 4th (257%), 16th (291%) and 48th (165%) week (P<0.0001). CYP2B6*6 genotype significantly influenced 4ß-OHC/Chol ratio at weeks 16 (P=0.02) and 48 (P=0.04) being highest in CYP2B6*6/*6>*1/*6>*1/*1. There were positive correlations between plasma efavirenz and 4ß-OHC/Chol ratios (week 4: P=0.02, week 16: P=0.001). CYP3A enzyme induction by efavirenz is pronounced in CYP2B6 slow metabolizers who have high efavirenz plasma exposure.

Effects of resource chemistry on the composition and function of stream hyporheic biofilms.

Fluvial ecosystems process large quantities of dissolved organic matter as it moves from the headwater streams to the sea. In particular, hyporheic sediments are centers of high biogeochemical reactivity due to their elevated residence time and high microbial biomass and activity. However, the interaction between organic matter and microbial dynamics in the hyporheic zone remains poorly understood. We evaluated how variance in resource chemistry affected the microbial community and its associated activity in experimentally grown hyporheic biofilms. To do this we fed beech leaf leachates that differed in chemical composition to a series of bioreactors filled with sediment from a sub-alpine stream. Differences in resource chemistry resulted in differences in diversity and phylogenetic origin of microbial proteins, enzyme activity, and microbial biomass stoichiometry. Specifically, increased lignin, phenolics, and manganese in a single leachate resulted in increased phenoloxidase and peroxidase activity, elevated microbial biomass carbon:nitrogen ratio, and a greater proportion of proteins of Betaproteobacteria origin. We used this model system to attempt to link microbial form (community composition and metaproteome) with function (enzyme activity) in order to better understand the mechanisms that link resource heterogeneity to ecosystem function in stream ecosystems.

Determining the time course of CYP3A inhibition by potent reversible and irreversible CYP3A inhibitors using A limited sampling strategy.

We established a new limited sampling strategy to assess CYP3A activity and evaluated the time course of reversible (voriconazole) and irreversible (ritonavir) CYP3A inhibition. In this randomized trial, two groups, each with eight healthy participants, received CYP3A inhibitors voriconazole or ritonavir orally for 9 days, with 3 mg midazolam (MDZ) administered before the inhibitor treatment, on days 1, 2, 3, 5, 8, and 9 during inhibitor treatment, and on days 10, 11, and 12 (3 days) after discontinuation. Plasma MDZ area under the curve (AUC) between 2 and 4 h after oral administration in the form of a solution strongly correlated with MDZ clearance. Using this parameter, maximum inhibition of voriconazole and ritonavir was calculated to have occurred only 48 h after starting of the inhibitor (percentage of baseline MDZ clearance, voriconazole: 10.6%; ritonavir: 8.4%). Recovery of CYP3A activity occurred with a half-life of 24 h after voriconazole, whereas ritonavir inhibition was still strong 3 days after discontinuation. These findings underscore the substantial and gradual alterations in dose requirements in the first days of and after such combination therapies.

Effect of rifampicin and CYP2B6 genotype on long-term efavirenz autoinduction and plasma exposure in HIV patients with or without tuberculosis.

We performed a prospective comparative study to examine, from a pharmacogenetics perspective, the effect of rifampicin (RIF) on long-term efavirenz (EFV) autoinduction and kinetics. In a study population of patients with HIV receiving EFV with RIF (arm 2, n = 54) or without RIF (arm 1, n = 128 controls), intraindividual and interindividual plasma EFV and 8-hydroxyefavirenz levels were compared at weeks 4 and 16 of EFV therapy. In arm 2, RIF was initiated 4 weeks before starting EFV. In controls (arm 1), the plasma EFV was significantly lower whereas 8-hydroxyefavirenz was higher at week 16 as compared to week 4. By contrast, there were no significant differences in plasma EFV and 8-hydroxyefavirenz concentrations over time in arm 2. At week 4, the plasma EFV concentration was significantly lower in arm 2 as compared to arm 1, but no significant differences were observed by week 16. When stratified by CYP2B6 genotype, significant differences were observed only with respect to CYP2B6*1/*1 genotypes. Ours is the first report of the CYP2B6 genotype-dependent effect of RIF on long-term EFV autoinduction.

Aumento de resistencia de Streptococcus agalactiae vaginal-anal en el tercer trimestre de gestación a eritromicina y clindamicina al cabo de una década de tamizaje universal / Streptococcus agalactiae increase in resistance to erythromycin and clindamycin in vaginal-anal colonization in third quarter of pregnancy in one decade of universal screening

Introduction: Streptococcus agalactiae (GBS) is the main causative agent of early perinatal sepsis. The acquisition of prevention policies has led to frequent use of intrapartum antibiotics. Surveillance of antimicrobial resistance is indispensable for defining drugs of choice and alternatives for such prophylaxis. Objectives: To determine the evolution of antimicrobial resistance of GBS from maternal colonization to drugs used in the prevention of neonatal sepsis, between 2002 and 2008. Methods: We studied 100 GBS positive vaginal and anal samples from pregnant women. Disc diffussion susceptibility method was performed for penicillin, ampicillin, cefazolin, erythromycin and clindamycin according to the Clinical and Laboratory Standards Institute (CLSI). Results: We analyzed the susceptibility of 99 strains. Seventeen were resistant to erythromycin (17.1 percent) and 13 were resistant to clindamycin (13.1 percent). Thirteen of the 17 strains resistant to erythromycin had the MLS phenotype (resistance to erythromycin and clindamycin) and 4 had the M phenotype (resistance to erythromycin only). Within the MLS phenotype, resistance was constitutive in 9 strains, and induced in 4 strains (positive D test). Compared with 2002 there was a significant increase in resistance to clindamycin (from 3.27 percent to 13.1 percent p < 0.002) and erythromycin (1.09 percent to 17 percent p < 0.001). 100 percent GSB remained sensitive to penicillin and ampicillin. Conclusions: GBS remains highly susceptible to drugs of choice for prevention of perinatal sepsis. There is a significant increase in antimicrobial resistance to clindamycin and erythromycin. Therefore, it is necessary to request susceptibility testing in GBS from third trimester of pregnancy screening in patients allergic to penicillin.

Introducción: Streptococcus agalactiae es el principal agente causal de sepsis perinatal precoz. La adquisición de políticas de prevención ha traído consigo la utilización frecuente de antimicrobianos intra-parto. La vigilancia de resistencia antimicrobiana se hace indispensable para definir el fármaco de elección y alternativas en dicha profilaxis. Nuestro centro realiza tamizaje universal desde hace 10 años. Objetivos: Determinar la evolución de la resistencia antimicrobiana de S. agalactiae de colonización materna, a los antimicrobianos utilizados en la prevención de sepsis neonatal, entre 2002 y 2008. Métodos: Se estudiaron 100 muestras vaginales-anales positivas para S. agalactiae de mujeres embarazadas, con edad gestacional de 35 a 37 semanas. Se realizó estudio de susceptibilidad in vitro por discos a penicilina, ampicilina, cefazolina, eritromicina y clindamicina, según método estandarizado por Clinical and Laboratory Standards Institute (CLSI). Resultados: Se analiza la susceptibilidad de 99 cepas. Diecisiete fueron resistentes a eritromicina (17,1 por ciento) y 13 eran resistentes a clindamicina (13,1 por ciento). De las 17 cepas resistentes a eritromicina, 13 eran fenotipo MLS y 4 del fenotipo M. Dentro del fenotipo MLS, la resistencia fue constitutiva en nueve cepas e inducible en cuatro cepas (test D positivo). En comparación con el año 2002, hubo un aumento significativo de resistencia a clindamicina (de 3,2 a 13,1 por ciento p < 0,002) y a eritromicina (de 1,09 a 17 por ciento p < 0,001). Streptococcus agalactiae se mantuvo 100 por ciento sensible a penicilina y ampicilina. Conclusiones: S. agalactiae mantiene alta sensibilidad a los antimicrobianos de elección para la prevención de sepsis neonatal y a un antimicrobiano alternativo: cefazolina. Se observó un aumento significativo de resistencia antimicrobiana a clindamicina y eritromicina. Se hace necesario, entonces, solicitar antibiograma en el tamizaje del tercer trimestre del embarazo, en pacientes alérgicas a penicilina.

[Tendon transfers for peroneal palsy - functional outcome]. / Motorische Ersatzplastiken zur Therapie der Peroneusparese - funktionelle Ergebnisse.

PURPOSE: Patients with persistent peroneal palsy may require treatment for gait disturbances if conservative treatment is not tolerated. Transfer of the tibialis posterior tendon can restore foot extension and improve the patients gait pattern. PATIENTS AND METHODS: Retrospective analysis (mean follow-up 40.8 months) of 13 patients. (7♀, 6♂; 1998-2005) after tibialis posterior tendon transfer through the interosseous membrane to the tibialis anterior and peroneus longus tendons. Evaluation focussed on hospitalisation periods, perioperative morbidity, functional outcome (range of motion, strength, pain and gait), return to work rate and self assessment with the Funktionsfragebogen Hannover (FFbH). RESULTS: 7 iatrogenic, 5 posttraumatic and 1 congenital peroneal palsy were treated. The patients presented after an average of 8.3 months (hospitalisation 11 days). Perioperative morbidity was 38.4% (15.4% reoperation rate). The mean active range of motion of the ankle was 3°/0°/56° (extension/flexion), the average strength was 3 (MRC) and pain was 4 (visual analogue scale 1-10). 12 patients were evaluated with normal or improved gait pattern without ortheses. The mean FFbH score was 70.9%. CONCLUSION: If conservative treatment for peroneal palsy fails to improve functional outcome tendon transfers such as the tibialis posterior tendon transfer are a considerable treatment option. However, significant morbidity rates have to be anticipated. Consecutive patients should be referred in good time to specialised units.

Optimizing Suprathel®-therapy by the use of Octenidine-Gel®.

OBJECTIVE: A prospective, randomized, non-blinded, clinical study was conducted to evaluate the feasibility and practicability as well as pain reduction and ease of handling of Flammazine® versus Octenidine-Gel® s a primary local antiseptic before synthetic skin substitute application in partial-thickness burns. METHODS: Thirty patients with a median age of 42 years suffering from second-degree burns were included in the study. Burns were randomly selected, one area was treated with Flammazine®/gauze, another area in the same patient was treated with Octenidine-Gel®/gauze as initial antiseptic treatment. Within 24 h the first gauze change was performed followed by wound inspection, disinfection and synthetic skin substitute application. The study focused on patient pain score, analysis of wound bed and ease of handling of the two local antiseptic agents. RESULTS: There was a significant difference between Flammazine® versus Octenidine-Gel® regarding patient pain score and ease of handling. Octenidine-Gel® was less painful (p < 0.05) and easier to handle (p < 0.05). There was no significant difference for wound bed evaluation between the two antiseptic agents. A tendency for better wound bed preparation was seen with the use of Octenidine-Gel®. CONCLUSION: Based on the findings of this study Octenidine-Gel® is recommended as a local antiseptic agent, because when compared to Flammazine®, Octenidine-Gel® proved to be better in terms of ease of care, simplicity application, with gentler and faster detachment of the gel from wound surfaces and consequently far less pain during dressing changes.

Long-term efavirenz autoinduction and its effect on plasma exposure in HIV patients.

We investigated the influence of gender and pharmacogenetic variations on long-term efavirenz autoinduction and disposition among patients with HIV in Tanzania (N = 129). Plasma concentrations (at 16 h) of efavirenz and 8-hydroxyefavirenz were quantified at weeks 4 and 16 of therapy. Genotyping was performed to identify cytochrome P450 (CYP) 2B6*6, CYP3A5*3, *6, and *7, and ABCB1-3435 C/T genotypes. There were reductions in the median efavirenz concentration (Wilcoxon matched-pair test P < 0.001) and efavirenz/8-hydroxyefavirenz ratio (P < 0.001) by 19 and 32%, respectively, at week 16 as compared with week 4. The proportion of patients with efavirenz concentration <1 µg/ml at week 16 was higher by 67, 25, and 5% in CYP2B6*1/*1, *1/*6, and *6/*6 genotypes, respectively. The defined therapeutic range based on observed plasma concentrations is affected by the time point of sampling and the CYP2B6 genotype. The effect of efavirenz autoinduction on reducing plasma exposure continues up to week 16 and predominantly affects CYP2B6 extensive metabolizers. Among CYP2B6 slow metabolizers, the presence of a CYP3A5 genotype allele is associated with greater effects of efavirenz autoinduction on plasma concentrations of the drug. The cumulative induction may influence the long-term antiretroviral therapy outcome, particularly in CYP2B6*1 carriers.

Matriderm® in depth-adjusted reconstruction of necrotising fasciitis defects.

Necrotising fasciitis (NF) and Fournier's gangrene are still a source of high morbidity and mortality and a significant health-care resource consumption. These difficult cases are increasingly being referred to burn centres for specialised wound and critical care issues. Besides the total body surface area (TBSA) affected, location, co-morbidities, age and an immediate surgical treatment are important prognostic factors. The treatment of these patients is challenging and best performed by prompt diagnosis, immediate radical surgical debridement and aggressive critical care management. Referral to a major burn centre may help provide optimal surgical intervention, wound care and critical care management.As soon as the patient is stabilised, reconstruction of the injured areas becomes the main focus. As often seen, complete loss of dermal structures needs a depth adjusted--'multilayer'--reconstruction especially in critical areas. In modern reconstructive surgery, concepts of layer-specific reconstruction, including dermal substitution have to be considered. In this article, we present our recent experiences of five patients with NF who underwent dermal reconstruction with Matriderm® not only for better skin quality but also in some cases as an alternative to flap surgery when joint capsules or tendons were exposed.

Topical application of cerium nitrate prevents burn edema after burn plasma transfer.

Thermal injuries of more than 20% body surface area (BSA) result in systemic capillary leakage with subsequent edema. This can similarly be induced by burn plasma transfer (BPT) from burned individuals to healthy rats. We evaluated if cerium nitrate (CN) bathing can prevent edema after BPT. Therefore, donor rats (DR) underwent thermal injury (100 degrees C water, 30%BSA, 12 s) for positive controls and were additionally bathed in CN (0.05M, at 10 and 120 min) for study groups. For negative controls DR underwent shamburn (37 degrees C water, 30%BSA, 12 s). DR-plasma (harvested 4 h post trauma) was transferred to healthy individuals. Intravital microscopy was performed in mesenteric venules (0/60/120 min). Edema was assessed by FITC-albumin extravasation. Additionally, leukocyte sticking (cells/mm(2)) and micro hemodynamic parameters were assessed. Significant systemic capillary leakage was observed after BPT at 120 min. Edema formation was significantly lower in negative controls. Topical CN application after 10 and 120 min reduced FITC-efflux to baseline levels. Adherent leukocytes increased slightly in all groups. Leukocyte-sticking tended to be reduced after CN bathing. In conclusion, BPT induces burn edema in healthy individuals. CN bathing after 10 and 120 min reduces mediator levels in burned individuals. Therefore, BPT after CN application does not induce burn shock anymore. Burn edema is partially independent from leukocyte activation because CN significantly influences macromolecular leakage whereas leukocyte activation is not significantly altered.

Stenotrophomonas maltophilia induced post-cataract-surgery endophthalmitis: Outbreak investigation and clinical courses of 26 patients.

BACKGROUND: Stenotrophomonas maltophilia, a microorganism which colonizes plastic material, is a rare causative agent of iatrogenic endophthalmitis. PATIENTS AND METHODS: A cluster of 26 cases of acute post-cataract-surgery endophthalmitis (PE) was identified. An outbreak investigation was performed. Information was abstracted from patients' charts and questionnaires sent to patients and their general practitioners. Vision was examined before, during, as well as one and six months after acute PE. Bacterial isolates were subjected to molecular typing. RESULTS: All patients initially received empiric systemic antibiotic treatment. The source of the infections was identified to be the rinsing solution used during cataract surgery, which was contaminated with two strains of S. maltophilia. Antibiotic therapy was subsequently changed to trimethoprim/sulfamethoxazol and ciprofloxacin for 30 days, complemented with iv fluocortolone and topical treatment with prednisolone, ciprofloxacin, and chloramphenicol. Twenty-one patients (81%) received pars plana vitrectomy and were additionally treated with intravitreal injections of vancomycin, amikacin and dexamethasone, or imipenem and dexamethasone, respectively. In addition, oxacillin, mezlocillin, and prednisolone were applied subconjunctivally after vitrectomy. Six months after acute infection, a final visual acuity of > or = 0.2 was achieved by 21/26 patients (80%), a visual acuity of > or = 0.5 by 14/26 patients (54%). Twenty of 26 patients (77%, 17 of whom had undergone vitrectomy) achieved a higher visual acuity than before surgery. Patients from the vitrectomy group had a median final visual acuity of 0.5 compared to 0.4 in the 5 patients without vitrectomy. There was 1 retinal ablation, 2 intra-retinal bleedings, and relapse of infection in 2/26 patients (8%), with isolation of S. maltophilia in one of the relapsing infection cases. CONCLUSIONS: Empiric antibiotic treatment of PE may not adequately treat rare pathogens such as S. maltophilia. Administration of an effective systemic or intravitreal antibiotic treatment after identification of S. maltophilia may have contributed to the favorable clinical course and relatively low relapse frequency in our patients. Despite the known problem of persistence of S. maltophilia, visual acuity outcome after treatment is comparable to PE induced by other Gram-positive or Gram-negative bacteria.

Abrogation of TGF-beta by antisense oligonucleotides modulates expression of VEGF and increases angiogenic potential in isolated fibroblasts from radiated skin.

The transforming growth factor-beta (TGF-beta) has been identified as an important component of wound healing. Recent developments in molecular therapy offer good prospects for the modulation of wound healing, specifically those targeting TGF-beta. The aim of this study was to analyze the effect of TGF-beta targeting on the expression of angiogenic vascular endothelial growth factor (VEGF), a key regulator of angiogenesis and in vitro angiogenic activity in fibroblasts isolated from radiation-induced chronic dermal wounds. The expression of angiogenic VEGF in tissue samples from radiation-induced chronic dermal wounds was investigated by immunohistochemistry and microarray technique. The effect of TGF-beta targeting using antisense oligonucleotides on the expression of VEGF in isolated fibroblasts was analyzed by ELISA and multiplex RT-PCR. Human endothelial cells (ECs) were grown in conditioned medium produced from the treated fibroblasts. EC migration was measured using a modified Boyden chamber; EC tube formation was analyzed under a light microscope. Immunohistochemical investigation and microarray analysis demonstrated a decreased expression of VEGF protein and mRNA in tissue samples from radiation-induced chronic dermal wounds compared to normal human skin. Antisense TGF-beta oligonucleotide treatment significantly up-regulated VEGF secretion in vitro. Addition of conditioned medium from TGF-beta antisense-treated fibroblasts resulted in an increase in EC cell migration and tube formation. In conclusion, our results demonstrate that TGF-beta antisense oligonucleotide technology may be a potential therapeutic option for stimulation of angiogenesis in radiation-induced dermal wounds.

TGF-beta1 antisense therapy modulates expression of matrix metalloproteinases in keloid-derived fibroblasts.

Transforming growth factor-beta1 (TGF-beta1) has been identified as an important regulator of wound healing. Recent developments in molecular therapy offer exciting prospects for the modulation of wound healing, specifically those targeting TGF-beta1. The purpose of this study was to analyze the effect of TGF-beta1 targeting on the expression of matrix metalloproteinases (MMPs) in fibroblasts cultured from earlobe keloids. The expression of MMP-2 and -9 in tissue samples from keloids was investigated by immunohistochemistry. The effect of TGF-beta1 targeting using antisense oligonucleotides on the expression of MMPs in keloid-derived fibroblasts was analysed by ELISA and multiplex RT-PCR. Immunohistochemical studies demonstrated an increased expression of MMP protein in tissue samples from keloids compared to normal human skin. Antisense TGF-beta1 oligonucleotide treatment significantly downregulated MMP-9 secretion in vitro. In conclusion, TGF-beta1 antisense oligonucleotide technology may be a potential therapeutic option for the inhibition of proteolytic tissue destruction in keloids.

[Usefulness of the determination of IgG and IgM antibodies by ELISA and immunocapture in a clinical series of human brucellosis]. / Utilidad de la determinación de anticuerpos IgG e IgM por ELISA e inmunocaptura en una serie clínica de brucelosis humana.

INTRODUCTION: The diagnostic difficulties of brucellosis makes the evaluation of new diagnostic tests necessary. OBJECTIVES: Evaluation of different commercial tests in the serological diagnosis of brucellosis by ELISA and immunocapture antibodies in a clinical series of patients with brucellosis of the Health Network of the Catholic University of Chile. METHODS: All the serums received in the Laboratory of Microbiology for suspicion of brucellosis during five years were studied. Two groups were obtained, one that fulfilled diagnostic criteria for brucellosis [clinical evidence, and/or positive blood culture and/or seroagglutination test (SAT) in titers > 1/160] and the control group. Each serum sample was analyzed using irnmunocapture-agglutination test (Brucellacapt), ELISA IgM and IgG. RESULTS: Of 10 patients with brucellosis, the serologic results were: 8/10 positives for ELISA IgG, 7/10 for Brucellacapt and SAT, and 5/10 for ELISA IgM. DISCUSSION: ELISA IgG alone was superior than SAT. The combination ELISA IgG/ Brucellacapt reaches the best detection performance (9/10) and can be an alternative to SAT.