Antifungal Efficacy during Candida krusei Infection in Non-Conventional Models Correlates with the Yeast In Vitro Susceptibility Profile

The incidence of opportunistic fungal infections has increased in recent decades due to the growing proportion of immunocompromised patients in our society. Candida krusei has been described as a causative agent of disseminated fungal infections in susceptible patients. Although its prevalence remains low among yeast infections (2-5%), its intrinsic resistance to fluconazole makes this yeast important from epidemiologic aspects. Non mammalian organisms are feasible models to study fungal virulence and drug efficacy. In this work we have used the lepidopteran Galleria mellonella and the nematode Caenorhabditis elegans as models to assess antifungal efficacy during infection by C. krusei. This yeast killed G. mellonella at 25, 30 and 37°C and reduced haemocytic density. Infected larvae melanized in a dose-dependent manner. Fluconazole did not protect against C. krusei infection, in contrast to amphotericin B, voriconazole or caspofungin. However, the doses of these antifungals required to obtain larvae protection were always higher during C. krusei infection than during C. albicans infection. Similar results were found in the model host C. elegans. Our work demonstrates that non mammalian models are useful tools to investigate in vivo antifungal efficacy and virulence of C. krusei. © 2013 Scorzoni et al.

Persistent efficacy of 3.5% doramectin compared to 3.15% ivermectin against gastrointestinal nematodes in experimentally-infected cattle in Brazil

The present study aimed to evaluate the persistent efficacy of a 3.5% doramectin* (700μg/kg) formulation compared to 3.15% ivermectin** (630μg/kg) treatment, administered subcutaneously at a dose of 1mL/50kg body weight in cattle experimentally infected with gastrointestinal nematodes. Seventy-two male crossbred Holstein cattle that were negative for helminth infection were divided into nine groups. Treatments of 3.5% doramectin (Groups 2, 4, 6 and 8) and 3.15% ivermectin (Groups 3, 5, 7 and 9) were administered on days 49, 42, 35 and 28 prior to challenge with infectious nematode larvae (L3). Animals in the control group (Group 1) received saline solution on day 49 before challenge. Beginning on day zero, each animal received 50mL orally of a mixed culture containing approximately 3,000 third stage larvae (L3) of Haemonchus (60%), Oesophagostomum (20%), Cooperia (15%) and Trichostrongylus (5%) for seven consecutive days, resulting in a total challenge of 21,000 larvae/animal. Due to the large number of cattle, autopsies were performed between days 28 and 35 after the last day of inoculation. The formulation containing doramectin (700 mcg/kg) achieved persistent efficacy against H. placei and C. punctata for 49 and 35days, respectively. The persistent efficacy of ivermectin (630 mcg/kg) against H. placei lasted for 49days, but this treatment was ineffective against C. punctata. Both formulations demonstrated persistent efficacy against T. axei for 49days. The persistent efficacy of doramectin (700 mcg/kg) and ivermectin (630 mcg/kg) lasted for 49 and 42days against O. radiatum, respectively. © 2012 Elsevier Ltd.

Efficacy of four irrigation needles in cleaning the apical third of root canals

This study aimed to evaluate the influence of irrigation needle gauge and design, and the final root canal diameter on the apical cleaning efficacy. Twelve human mandibular incisors were used. At different stages of root canal widening (sizes 20, 30 and 40 K-files), root canals were filled with radiopaque contrast medium. Four different needles were evaluated: 23G with side opening, 22G with apical opening, 30G with side opening and 30G with apical opening. Irrigation was carried out with 2 mL distilled water. The same tooth was radiographed with a digital system several times to assess the four types of needle in those three stages of canal widening. Pre-irrigation (canals filled with contrast) and post-irrigation (canals with remaining contrast) images were submitted to digital subtraction using the Adobe Photoshop CS4 program. Pre-irrigation (filled with contrast) and subtracted (cleaned by irrigation) areas were outlined by a trained and blinded operator using the Image Tool 3.0 software. Their ratio was calculated to express the percentage of apical cleaning in each stage of canal widening (sizes 20, 30 and 40 K-files) with each of the four needles. Data obtained were subjected to one-way ANOVA and Tukey's tests. The 30G needles with side and apical opening promoted better apical cleaning at all stages of root canal widening (p<0.05). In conclusion, smaller diameter needles were more efficacious in cleaning the apical third of the root canals, regardless of their design.

Evaluation of a propolis water extract using a reliable RP-HPLC methodology and in vitro and in vivo efficacy and safety characterisation

Since the beginning of propolis research, several groups have studied its antibacterial, antifungal, and antiviral properties. However, most of these studies have only employed propolis ethanolic extract (PEE) leading to little knowledge about the biological activities of propolis water extract (PWE). Based on this, in a previous study, we demonstrated the anti-inflammatory and immunomodulatory activities of PWE. In order to better understand the equilibrium between effectiveness and toxicity, which is essential for a new medicine, the characteristics of PWE were analyzed. We developed and validated an RP-HPLC method to chemically characterize PWE and PEE and evaluated the in vitro antioxidant/antimicrobial activity for both extracts and the safety of PWE via determining genotoxic potential using in vitro and in vivo mammalian micronucleus assays. We have concluded that the proposed analytical methodology was reliable, and both extracts showed similar chemical composition. The extracts presented antioxidant and antimicrobial effects, while PWE demonstrated higher antioxidant activity and more efficacious for the most of the microorganisms tested than PEE. Finally, PWE was shown to be safe using micronucleus assays. © 2013 Bruno Alves Rocha et al.

The efficacy of the self-adjusting file and ProTaper for removal of calcium hydroxide from root canals

Objective: The goal of this study was to evaluate the efficacy of the Self-Adjusting File (SAF) and ProTaper for removing calcium hydroxide [Ca(OH)2] from root canals. Material And Methods: Thirty-six human mandibular incisors were instrumented with the ProTaper system up to instrument F2 and filled with a Ca(OH)2-based dressing. After 7 days, specimens were distributed in two groups (n=15) according to the method of Ca(OH)2 removal. Group I (SAF) was irrigated with 5 mL of NaOCl and SAF was used for 30 seconds under constant irrigation with 5 mL of NaOCl using the Vatea irrigation device, followed by irrigation with 3 mL of EDTA and 5 mL of NaOCl. Group II (ProTaper) was irrigated with 5 mL of NaOCl, the F2 instrument was used for 30 seconds, followed by irrigation with 5 mL of NaOCl, 3 mL of EDTA, and 5 mL of NaOCl. In 3 teeth Ca(OH)2 was not removed (positive control) and in 3 teeth canals were not filled with Ca(OH)2 (negative control). Teeth were sectioned and prepared for the scanning electron microscopy. The amounts of residual Ca(OH)2 were evaluated in the middle and apical thirds using a 5-score system. Results: None of the techniques completely removed the Ca(OH)2 dressing. No difference was observed between SAF and ProTaper in removing Ca(OH)2 in the middle (P=0.11) and the apical (P=0.23) thirds. Conclusion: The SAF system showed similar efficacy to rotary instrument for removal of Ca(OH)2 from mandibular incisor root canals.

Liposomal-praziquantel: Efficacy against Schistosoma mansoni in a preclinical assay

Currently, schistosomiasis mansoni is treated clinically with praziquantel (PZQ). Nevertheless, cases of tolerance and resistance to this drug have been reported, creating the need to develop new drugs or to improve existing drugs. Considering the small number of new drugs against Schistosoma mansoni, the design of nanotechnology-based drug delivery systems is an important strategy in combating this disease. The aim of this study was to evaluate the activity of PZQ containing liposome (lip.PZQ) on S. mansoni, BH strain. Mice were treated orally with different concentrations of PZQ and lip.PZQ 30 and 45 days following infection. The number of worms, recovered by perfusion of the hepatic portal system, and the number of eggs found in the intestine and liver were analysed. Parasite egg counts were also performed. The most active formulation for all parameters was 300. mg/kg of lip.PZQ, since as it decreased the total number of worms by 68.8%, the number of eggs in the intestine by 79%, and the number of hepatic granulomas by 98.4% compared to untreated controls. In addition, this concentration decreased egg counts by 55.5%. The improved efficacy of the treatment with lip.PZQ, especially when administered 45 days following infection, compared with the positive-control group (untreated) and the groups that received free PZQ, can be explained by greater bioavailability in the host organism; the preferred target of lip.PZQ is the liver, and lip.PZQ is better absorbed by the tegument of S. mansoni, which has an affinity for phospholipids. © 2013 Elsevier B.V.

Anthelmintic efficacy of an oral formulation of Aurixazol against gastrointestinal nematodes of naturally and experimentally infected sheep

As a result of the need to develop new active principles for the control of endoparasites in ruminants, the present in vivo study evaluated a formulation containing 24% Aurixazol (48 mg/kg), a parasiticide molecule based on disophenolate of levamisole. Two experiments were conducted: one evaluating the anthelmintic efficacy of 24% Aurixazol (48 mg/kg) against gastrointestinal nematodes in naturally infected sheep, compared to an association of ivermectin (0.2 mg/kg) + albendazole (5.0 mg/kg) + levamisole (7.5 mg/kg) (IAL), and a second one which evaluated the persistent efficacy of the same formulation against immature stages (L4) and adults of Haemonchus contortus in experimentally infected animals. In experiment I, against H. contortus, the formulation of Aurixazol and the IAL association reached efficacies (arithmetic means) of 99.32% and 96.11%, respectively. For Trichostrongylus colubriformis, the efficacy values were 88.92% and 98.08% for Aurixazol and the IAL association, respectively. Both formulations were totally effective against Oesophagostomum columbianum (100%). The results of the statistical analysis demonstrated that the mean parasitic burden of treated animals was significantly different (P ≤ 0.05) compared to the average number of helminths diagnosed in animals from the control group for H. contortus, T. colubriformis and O. columbianum. Comparing only the treated groups, it was possible to verify that the average number of H. contortus recovered from animals treated with Aurixazol was different (P ≤ 0.05) when compared to the mean amount recovered from sheep treated with the IAL association. When evaluating the prevention of H. contortus infection in experiment II, Aurixazol did not present preventive efficacy. Up until 21 days after treatment the groups treated with Aurixazol contained less adults and L4 of H. contortus (P ≤ 0.05) when compared to the non-medicated control group. However, future studies will be necessary to assess the effectiveness of Aurixazol against nematode strains resistant to levamisole and disophenol, but the efficacy results described in this study allow to state that Aurixazol can, associated with other measures, become an important tool in the control of sheep nematodes. © 2013.

Reinforcing production cooperation and dialogue spaces: the role of SMEs

Foreword by Alicia Bárcena and Jorge Valdez

Efficacy of the d-phenothrin/pyriproxyfen association against mites in naturally co-infested rabbits

O objetivo do presente estudo foi avaliar a eficácia da associação de d-fenotrina e piriproxifen no controle de infestações simultâneas por Psoroptes ovis, Cheyletiella parasitivorax e Leporacarus gibbus em coelhos naturalmente co-infestados. Vinte coelhos mestiços (Nova Zelândia Branco x Califórnia) infestados simultaneamente pelas três espécies de ácaros foram divididos aleatoriamente em dois grupos. Todos os coelhos infestados por apresentavam eritema, hiperemia e formação de crostas nas orelhas, causados por P. ovis. Infestações simultâneas por C. parasitivorax e L. gibbus foram considerados assintomáticas em todos os animais. Dez animais foram tratados com uma formulação spray contendo d-fenotrina a 4,4% e piriproxifen a 0,148%, pulverizando toda a superfície corporal de forma uniforme, incluindo a face interna das orelhas. Os outros 10 coelhos não foram t ratados, sendo mantidos como grupo controle. Os animais foram avaliados nos dias 7, 14, 21, 28 e 35 pós-tratamento. A associação de d-fenotrina e piriproxifen foi 100% eficaz no controle das três espécies de ácaros e foi responsável pela remissão das lesões de sarna psoróptica nos animais tratados. Não foram observados sinais de intoxicação. Os resultados indicam que a formulação spray de d-fenotrina e piriproxifen em uma única aplicação é uma opção clinicamente segura e eficaz no controle de infestações por ácaros em coelhos.

Efficacy of an Oral Appliance for the Treatment of Obstructive Sleep Apnea

Purpose: The aim of this study was to validate the use of a mandibular repositioner appliance (MRA) to treat obstructive sleep apnea (OSA) and primary snoring, comparing polysomnographic and Epworth Sleepiness Scale (ESS) data obtained prior to and during MRA treatment. Materials and Methods: Sixty-three patients who presented with different degrees of OSA severity or primary snoring were fitted to a PM positioner between 2009 and 2011. The diagnosis was established by a polysomnogram (PSG) prior to treatment and after 6 months to verify the efficacy of MRA therapy. Subjective daytime sleepiness was evaluated by ESS questionnaire prior to treatment and at the follow-up. Results: Patients were divided into primary snoring and OSA groups. For the primary snoring group, PSG variables did not show significant results, except for a decrease in snoring. For the OSA group, the mean apnea-hypopnea index (AHI) was reduced from 23.0 +/- 11 to 5.3 +/- 4.0 and median ESS reduced significantly from 13.0 to 8.5. Complete response (AHI < 5) was found in 25 (40%) patients and partial response (AHI <= 10) in 27 (43%) patients. Conclusion: The findings validate the efficacy of the adjustable PM positioner for the safe treatment of OSA.

In vitro evaluation of permeation, toxicity and effect of praziquantel-loaded solid lipid nanoparticles against Schistosoma mansoni as a strategy to improve efficacy of the schistosomiasis treatment

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efficacy of sperm motility after processing and incubation to predict pregnancy after intrauterine insemination in normospermic individuals

Background: Intrauterine insemination (IUI) is widely used to treat infertility, and its adequate indication is important to obtain good pregnancy rates. To assess which couples could benefit from IUI, this study aimed to evaluate whether sperm motility using a discontinuous gradient of different densities and incubation in CO2 in normospermic individuals is able to predict pregnancy.Methods: A total of 175 couples underwent 175 IUI cycles. The inclusion criteria for women were as follows: 35 years old or younger (age range: from 27 to 35 years) with normal fallopian tubes; endometriosis grades I-II; unexplained infertility; nonhyperandrogenic ovulatory dysfunction. Men with normal seminal parameters were also included. All patients underwent ovarian stimulation with clomiphene citrate and human hMG or r-FSH. When one or (at most) three follicles measuring 18 to 20 mm were observed, hCG (5000 UI) or r-hCG (250 mcg) was administered and IUI performed 36-40 h after hCG. Sperm processing was performed using a discontinuous concentration gradient. A 20 microliters aliquot was incubated for 24 h at 37 degrees C in 5% CO2 following a total progressive motility analysis. The Mann-Whitney and Chi-square tests, as well as a ROC curve were used to determine the cutoff value for motility.Results: Of the 175 couples, 52 (in 52 IUI cycles) achieved clinical pregnancies (CP rate per cycle: 29.7%). The analysis of age, duration and causes of infertility did not indicate any statistical significance between pregnancy and no pregnancy groups, similar to the results for total sperm count and morphology analyses, excluding progressive motility (p < 0.0001). The comparison of progressive motility after processing and 24 h after incubation between these two groups indicated that progressive motility 24 h after incubation was higher in the pregnancy group. The analysis of the progressive motility of the pregnancy group after processing and 24 h after incubation has not shown any motility difference at 24 h after incubation; additionally, in couples who did not obtain pregnancy, there was a statistically significant decrease in progressive motility 24 h after incubation (p < 0.0001). The ROC curve analysis generated a cutoff value of 56.5% for progressive motility at 24 h after incubation and this cutoff value produced 96.1% sensitivity, 92.7% specificity, 84.7% positive predictive value and 98.3% negative predictive value.Conclusions: We concluded that the sperm motility of normospermic individuals 24 h after incubation at 37 degrees C in 5% CO2, with a cutoff value of 56.5%, is predictive of IUI success.

Efficacy of Surface Treatments on the Bond Strength of Resin Cements to Two Brands of Zirconia Ceramic

Purpose: To compare the shear bond strength (SBS) of two cements to two Y-TZP ceramics subjected to different surface treatments.Materials and Methods: Zirconia specimens were made from Lava (n = 36) and IPS e.max ZirCAD (n = 36), and their surfaces were treated as follows: no treatment (control), silica coating with 30-mu m silica-modified alumina (Al2O3) particles (CoJet Sand), or coating with liners Lava Ceram for Lava and Intensive ZirLiner for IPS e.max ZirCAD. Composite resin cylinders were bonded to zirconia with Panavia F or RelyX Unicem resin cements. All specimens were thermocycled (6000 cycles at 5 degrees C/55 degrees C) and subjected to SBS testing. Data were analyzed by post-hoc test Tamhane T2 and Scheffe tests (alpha = 0.05). Failure mode was analyzed by stereomicroscope and SEM.Results: With both zirconia brands, CoJet Sand showed significantly higher SBS values than control groups only when used with RelyX Unicem (p = 0.0001). Surface treatment with liners gave higher SBS than control groups with both ceramic brands and cements (p < 0.001). With both zirconia brands, the highest SBS values were obtained with the CoJet and RelyX Unicem combination (> 13.47 MPa). Panavia F cement showed significantly better results when coupled with liner surface treatment rather than with CoJet (p = 0.0001, SBS > 12.23 MPa). In untreated controls, Panavia F showed higher bond strength than RelyX Unicem; the difference was significant (p = 0.016) in IPS e.max ZirCAD. The nontreated specimens and those treated with CoJet Sand exhibited a high percentage of adhesive and mixed A (primarily adhesive) failures, while the specimens treated with liners presented an increase in mixed A and mixed C (primarily cohesive) failures as well as some cohesive failure in the bulk of Lava Ceram for both cements.Conclusion: CoJet Sand and liner application effectively improved the SBS between zirconia and luting cements. This study suggests that different interactions between surface treatments and luting cements yield different SBS: in clinical practice, these interactions should be considered when combining luting cements with surface treatments in order to obtain the maximum bond strength to zirconia restorations.

Concentrations of and application protocols for hydrogen peroxide bleaching gels: Effects on pulp cell viability and whitening efficacy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)