Antibacterial activity, cytocompatibility and effect of Bio-C Temp bioceramic intracanal medicament on osteoblast biology.

AIM: To analyse the antimicrobial and biological properties of a new bioceramic intracanal medicament (Bio-C Temp), and to compare it with two calcium hydroxide-based intracanal medicaments (Calen® and UltraCal® XS). METHODOLOGY: The direct contact and the crystal violet tests were performed to assess the antimicrobial activity of intracanal medicaments against Enterococcus faecalis. The cytocompatibility and the effect of the medication on the biology of the human osteoblast-like cell line (Saos-2) were evaluated with methylthiazole tetrazolium (MTT), neutral red, alkaline phosphatase activity and mineralization (alizarin red) assays. The data were analysed using one-way anova and Tukey's tests, two-way anova and Bonferroni's tests, or Kruskal-Wallis and Dunn's tests (α = 0.05). RESULTS: Bio-C Temp had significantly less antibacterial activity and biofilm biomass reduction than the other intracanal medicaments (P < 0.05). There was no difference in the viability of Saos-2 exposed to the various intracanal medicaments, except regarding the 1 : 2 dilution, when the Bio-C Temp group had significantly lower cell viability than the UltraCal® XS and Calen® groups (P < 0.05). Bio-C Temp induced significantly greater ALP activity than the other intracanal medicaments (P < 0.05) at day 1. Calen® induced significantly greater deposition of mineralized nodules than the other intracanal medicaments (P < 0.05), and no difference was observed between Bio-C Temp and UltraCal® XS (P > 0.05). CONCLUSIONS: Bio-C Temp had similar cytocompatibility at higher dilutions, and higher or similar induction of ALP activity and deposition of mineralized nodules in comparison with Calen® and UltraCal® XS. However, it had significantly less antibacterial and antibiofilm activity than Calen® and UltraCal® XS.

Effects of octenidine applied alone or mixed with sodium hypochlorite on eukaryotic cells.

AIM: To assess the effects of octenidine dihydrochloride (OCT) on eukaryotic cells and the cytotoxicity of OCT associated with sodium hypochlorite - NaOCl (NaOCl/OCT). METHODOLOGY: L929 fibroblasts and human osteoblast-like cells (Saos-2) were exposed to 0.1% OCT, 2% CHX, 2.5% NaOCl, 5.25% NaOCl and mixtures of 5.25% NaOCl and 0.1% OCT (NaOCl/OCT) at 90 : 10, 80 : 20 and 50 : 50 ratios. Cell viability was assessed by methyl-thiazol-tetrazolium (MTT) and neutral red (NR) assays; type of cell death, by flow cytometry; cytoskeleton, by actin and α-tubulin fluorescence; and alkaline phosphatase (ALP) activity, by thymolphthalein release. The data were analysed by two-way ANOVA and Bonferroni tests (α = 0.05). RESULTS: MTT and NR assays revealed that 0.1% OCT had the lowest cytotoxicity (P < 0.05), followed by 2% CHX (P < 0.05). The 2.5% NaOCl, NaOCl/OCT 80 : 20 and NaOCl/OCT 50 : 50 solutions had intermediate cytotoxicity. NaOCl 5.25% and NaOCl/OCT 90 : 10 had the highest cytotoxicity (P < 0.05). The OCT group had a higher percentage of viable cells than the NaOCl and CHX groups (P < 0.05), and induced apoptosis at higher doses. The cytoskeleton alterations were observed at 0.12%, 0.6% and 2.02% for the NaOCl, CHX and OCT groups, respectively. The solutions did not induce ALP activity. CONCLUSION: Octenidine dihydrochloride was less cytotoxic, induced apoptosis at higher doses, caused few changes in the cytoskeleton and did not induce alkaline phosphatase activity. In addition, octenidine dihydrochloride reduced the cytotoxicity of 5.25% NaOCl when combined at 20 and 50%.

Effect of immersion in distilled water or phosphate-buffered saline on the solubility, volumetric change and presence of voids within new calcium silicate-based root canal sealers.

AIM: To assess the effect of immersion in distilled water or phosphate-buffered saline (PBS) on the solubility, volumetric change and presence of voids of calcium silicate-based root canal sealers (TotalFill BC, Sealer Plus BC and Bio-C), in comparison with the gold standard epoxy resin-based sealer (AH Plus). METHODOLOGY: All properties were evaluated after immersion in distilled water or PBS. Solubility was determined by the percentage of mass loss, whereas volumetric change and presence of voids were evaluated by micro-computed tomography, after 7 days of immersion. The volumetric change and percentage of voids between the baseline (after setting) and the experimental period were calculated. Statistical analysis was performed using one-way anova and Tukey's or Student's t-tests (α = 0.05). RESULTS: The calcium silicate-based sealers had significantly greater solubility and volumetric loss than AH Plus, after immersion in distilled water or PBS (P < 0.05). Bio-C had the greatest solubility (P < 0.05), followed by TotalFill BC and Sealer Plus BC, which were similar (P > 0.05). Regarding the volumetric change, AH Plus had a volume increase, with similar values in distilled water and PBS (P > 0.05). TotalFill BC, Sealer Plus BC and Bio-C had a similar volumetric change (P > 0.05). The calcium silicate-based materials had the greatest solubility and volume loss after immersion in distilled water (P < 0.05). There was no difference in the percentage of voids amongst the sealers, before and after immersion in distilled water or PBS (P > 0.05). CONCLUSIONS: TotalFill BC, Sealer Plus BC and Bio-C had significantly greater solubility and volumetric loss than AH Plus. Although storage in PBS significantly reduced the solubility and volumetric change of calcium silicate-based sealers, their solubility remained above that recommend by ISO 6876. All the sealers evaluated had low and similar voids, even after immersion in distilled water or PBS.

Cytocompatibility, bioactive potential and antimicrobial activity of an experimental calcium silicate-based endodontic sealer.

AIM: To evaluate the cytocompatibility, bioactive potential, antimicrobial and antibiofilm activities of an experimental calcium silicate-based endodontic sealer, in comparison with TotalFill BC Sealer and AH Plus. METHODOLOGY: Cytocompatibility was assessed by methyltetrazolium (MTT) and neutral red (NR) assays, after exposure of the Saos-2 cells to the sealer extracts (1 : 2, 1 : 4, 1 : 8, 1 : 16 and 1 : 32 dilutions) for 24 h. The sealers were manipulated and placed in 12-well culture plates and exposed to ultraviolet light; then, 5 mL of DMEM without serum was added. Cell bioactivity was evaluated by alkaline phosphatase activity (ALP) and Alizarin red staining (ARS). Antimicrobial and antibiofilm activities were evaluated by direct contact test (DCT) on planktonic cells (DCTPC) and modified DCT on biofilm formed in bovine dentine blocks (MDCT). MTT, NR and ALP data were analysed by two-way anova and Bonferroni tests; ARS data by anova and Tukey's tests; and the microbiological data by Kruskal-Wallis and Dunn tests (α = 0.05). RESULTS: The experimental sealer, TotalFill BC and AH Plus were not cytotoxic to Saos-2, in comparison with the negative control (P > 0.05). Greater ALP was observed after 7 days of exposure of Saos-2 to AH Plus and the experimental sealer (P < 0.05) when compared to the control. Significantly greater mineralized nodule production was observed for TotalFill BC and the experimental sealer (P < 0.05). In DCTPC, the experimental sealer and TotalFill BC were associated with a significantly greater reduction of E. faecalis (P < 0.05) and eliminated C. albicans. In MDCT, the experimental sealer and TotalFill BC had significantly greater antibiofilm efficacy (P < 0.05). CONCLUSIONS: The experimental calcium silicate-based sealer was cytocompatible, bioactive, antimicrobial against E. faecalis and C. albicans and effective against E. faecalis biofilms, with potential for use in root canal treatment.

Cellular and extracellular vesicular origins of miRNAs within the bovine ovarian follicle.

The ovarian follicle components must provide an ideal environment to ensure the success of reproductive processes, and communication between follicular cells is crucial to support proper oocyte growth. Recently, it has been demonstrated that the presence of extracellular vesicles (EVs) carrying microRNAs (miRNAs) in follicular fluid represents an important autocrine and paracrine communication mechanism inside the ovarian follicle. In this study, we tested the hypothesis that the miRNA content of EVs isolated from ovarian follicular (granulosa and cumulus-oocyte complexes) cell-conditioned culture media is dependent upon cell type. We initially screened bovine granulosa cells (GCs) and cumulus-oocyte complexes (COCs), as well as their derived EVs for 348 miRNAs using real-time PCR, and detected 326 miRNAs in GCs and COCs cells and 62 miRNAs in GCs and COCs EVs. A bioinformatics analysis of the identified cell-specific and differentially expressed miRNAs predicted that they likely modulate important cellular processes, including signalling pathways such as the PI3K-Akt, MAPK and Wnt pathways. By investigating the origins of miRNAs within the follicular fluid, the results of this study provide novel insights into follicular miRNA content and intercellular communication that may be of invaluable use in the context of reproductive technologies, diagnostic of ovarian-related diseases and/or the identification of biomarkers for oocyte and embryo quality.

Effectiveness of several solutions to prevent the formation of precipitate due to the interaction between sodium hypochlorite and chlorhexidine and its effect on bond strength of an epoxy-based sealer.

AIM: To evaluate the effectiveness of isopropyl alcohol, saline or distilled water to prevent the precipitate formed between sodium hypochlorite (NaOCl) and chlorhexidine (CHX) and its effect on the bond strength of an epoxy-based sealer in radicular dentine. METHODOLOGY: The root canals of 50 extracted human canines (n = 10) were instrumented. In G1, root canals were irrigated with 17% EDTA and 2.5% NaOCl; G2, as G1, except that 2% CHX was used as the final irrigant. In the other groups, intermediate flushes with isopropyl alcohol (G3), saline (G4) or distilled water (G5) were used between NaOCl and CHX. The specimens were submitted to SEM analysis to evaluate the presence of debris and smear layer, in the apical and cervical segments. In sequence, fifty extracted human canines were distributed into five groups (n = 10), similar to the SEM study. After root filling, the roots were sectioned transversally to obtain dentine slices, in the cervical, middle and apical thirds. The root filling was submitted to a push-out bond strength test using an electromechanical testing machine. Statistical analysis was performed using Kruskal-Wallis and Dunn's tests (α = 5%). RESULTS: All groups had similar amounts of residue precipitated on the canal walls (P > 0.05). The push-out bond strength values were similar for all groups, independently of the root third evaluated (P > 0.05). CONCLUSIONS: Isopropyl alcohol, saline and distilled water failed to prevent the precipitation of residues on canal walls following the use of NaOCl and CHX. The residues did not interfere with the push-out bond strength of the root filling.

Congenital toxoplasmosis in the state of Minas Gerais, Brazil: a neglected infectious disease?

This study aimed to investigate the distribution of congenital toxoplasmosis in the state of Minas Gerais, Southeastern Brazil and describe the demographic and socioeconomic profile of the municipalities associated with the disease. An ecological study was conducted using socioeconomic indicators of a database (MGSSRI) created by Fundação João Pinheiro (a government technical support agency of Minas Gerais), in order to show the development of the municipalities in the state. The prevalence of toxoplasmosis was the outcome and the items of the MGSSRI were the explanatory variables. Of 146,307 newborns screened (November 2006 to May 2007), 190 had congenital toxoplasmosis, yielding a prevalence of 1·3/1000, ranging from 0 to 76·9/1000 in the municipalities. The multivariate model indicated a higher occurrence of toxoplasmosis in municipalities with smaller populations and worse indexes of tax performance. Congenital toxoplasmosis appears to be a neglected disease in the state of Minas Gerais, given the high prevalence found and its concentration in municipalities with worse socioeconomic indexes.

Obstructive sleep apnea and biomarkers of inflammation in ischemic stroke.

BACKGROUND AND OBJECTIVES: Despite a confirmed association between obstructive sleep apnea (OSA) and stroke, the pathogenesis of OSA in stroke has not been hitherto clarified. The aim of this study was to evaluate the relationship between respiratory abnormalities and atherogenic pro-inflammatory markers, interleukin-1beta (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) in acute ischemic stroke patients. METHODS: Nocturnal polygraphy was performed in 50 consecutive patients with acute ischemic stroke in the first week after the event. Levels of inflammatory markers (IL-6, IL-1ß and TNF-α) were determined from morning blood samples and comparatively analyzed between cases with and without severe OSA and with age-matched controls. RESULTS: All patients with acute ischemic stroke, 31 men, mean age (64.3 ± 7.7 years), had apnea-hypopnea index (AHI) > 5 and 35 (70%) had severe OSA (AHI ≥ 30). Hypertension was more frequent in patients with severe OSA (85.7%) when compared to controls (40.0%) (P = 0.002). Stroke outcome, assessed by the Barthel index, tended to be more severe (P = 0.06) in cases with severe OSA. Patients with mild/moderate OSA and with severe OSA showed higher levels of IL-6 when compared to controls (P = 0.01 and P = 0.000, respectively). Among cases with acute stroke and severe OSA, IL-6 levels were correlated with lower oxyhemoglobin desaturation (r=-0.30; P = 0.001) and with the desaturation index (r = 0.15; P = 0.02). CONCLUSIONS: IL-6, an atherogenic marker, shows a commensurate increase in stroke patients with OSA. It is correlated with oxyhemoglobin desaturation and with desaturation index and may be a surrogate measure to evaluate continuous positive airway pressure therapy.

Antibacterial effectiveness of several irrigating solutions and the Endox Plus system - an ex vivo study.

AIM: To compare the ex vivo antibacterial effectiveness of the Endox Plus system and sodium hypochlorite (NaOCl) in combination with BioPure MTAD (Tulsa Dental, Tulsa, OK, USA) or with EDTA in Enterococcus faecalis-contaminated root canals. METHODOLOGY: After initial preparation, the root canals of 70 single-rooted human teeth were inoculated with E. faecalis (ATCC 29212) and incubated for 21 days. Specimens were divided into five groups: Endox Plus/saline; 2.5% NaOCl/MTAD; 2.5% NaOCl/EDTA; saline (positive control); negative control (root canals not prepared, nor irrigated). Samples were collected using paper points. Microbiological analysis evaluated the number of CFUs. Data were analysed by anova and Tukey tests at 0.05 significance. RESULTS: All specimens had bacterial growth after the incubation period, with similar CFU per mL counts (P > 0.05). After chemo-mechanical preparation, the number of bacteria in all groups reduced, except for the negative control. No significant differences were observed between 2.5% NaOCl/MTAD and 2.5% NaOCl/EDTA, but these groups had lower CFU counts than the other groups (P < 0.05). In the final samples, an increase in the bacterial counts was observed for Endox Plus/saline, 2.5% NaOCl/MTAD, 2.5% NaOCl/EDTA and saline (P < 0.05) with no significant differences between these groups. CONCLUSIONS: This ex vivo study revealed that the Endox Plus system was associated with a reduced antibacterial effectiveness compared with conventional irrigation using 2.5% NaOCl/MTAD and 2.5% NaOCl/EDTA. All irrigation procedures allowed recovery of bacteria 7 days after treatment, demonstrating persistence of contamination within the root canal system.

Shortage of Donors for Intestinal Transplantation in São Paulo, Brazil.

BACKGROUND: The number of deceased organ donors has decreased slightly over the past 4 years. Although the pool of intestinal transplantation candidates is relatively small, donor allocation is challenging because of the inability to maintain the donor in a good condition and the complexities involved in making a suitable weight match between donors and recipients. Our goal was to analyze the epidemiologic profile of potential donors based on the organs offered by the regional Organ Procurement Organization from Hospital das Clinicas-USP (OPO/HC-USP) and attempt to estimate possible matches and program viability. METHODS: We retrospectively analyzed information from the OPO/HC-USP database regarding organs offered over the past 7 years as well as patients listed in our program. Data were collected regarding donor characteristics (eg, sex, age, race, body mass index, blood type, cause of death) and medical care details (eg, intensive care unit stay, use of vasopressor agents and antibiotics). RESULTS: In this time period, there were 18,103 brain death notifications in the state of São Paulo; 5,202 (35%) became viable donors, resulting in 5,201 (99%) effectively used livers and kidneys. Most potential donors were male, in their 40s, white, and had blood type O. Only 3 potential donors from OPO/HC-USP would have reached the established minimum criteria for intestinal donation over these 7 years.

L-Alanyl-Glutamine Attenuates Oxidative Stress in Liver Transplantation Patients.

BACKGROUND: Ischemia/reperfusion injury during liver transplantation can cause severe damage to the graft. The objective of this randomized, double-blind study was to evaluate the possible protective effects of L-alanyl-glutamine on the liver graft. METHODS: The sample included 33 patients from a liver transplantation service in Northeastern Brazil. Before cold ischemia, the patients received 50 g of L-alanyl-glutamine (treatment group) or saline (control group) through the portal vein. The graft was biopsied at the time of recovery, at the beginning of warm ischemia, and at the end of transplantation to determine malondialdehyde (MDA), heat-shock protein (Hsp)70, nuclear factor kappa-beta (NFkB), superoxide dismutase (SOD), and reduced glutathione (GSH) levels. RESULTS: The blood parameters were similar in the two groups. In the treatment group, MDA did not increase at the beginning of cold ischemia and decreased at the end of transplantation. This phenomenon was not observed in the control group. GSH, SOD, Hsp70, and NFkB levels were similar in the two groups. CONCLUSIONS: Our findings suggest that preconditioning with L-alanyl-glutamine attenuates the effects of ischemia/reperfusion-related oxidative stress and reduces lipid peroxidation in the grafts of liver transplantation patients.

In vitro human immune reactivity of fast protein liquid chromatography fractionated Paracoccidioides brasiliensis soluble antigens.

Soluble antigens of Paracoccidioides brasiliensis yeast cells (PbAg) were fractionated in a fast protein liquid chromatography (FPLC) system, using Q-Sepharose anion-exchange resin, in order to characterize antigenic fractions that could elicit cell reactivity and antibody recognition in human paracoccidioidomycosis (PCM). PbAg fractions were eluted by 20 mM Tris-HCl solution (pH 9.6) with an increasing gradient up to 1 M NaCl. The FPLC system was able to resolve 7 fractions, enumerated from 0 to VI, according to the elution on the NaCl gradient. The analysis of each fraction on SDS-PAGE showed that fractions 0 to V were constituted by multiple protein bands with molecular mass ranging from 18 to 114 kDa. Large amounts of nucleic acids were evidenced in fraction VI, as revealed by agarose gel stained with ethidium bromide. Sera from PCM patients presenting different clinical forms contained antibodies that recognized antigens in all fractions with the exception of fraction VI as detected by ELISA. Further studies were designed to investigate the capacity of these fractions to induce cell proliferation. It was demonstrated that fractions III and V (200 and 450 mM NaCl, respectively) stimulated a significant proliferative response of peripheral blood mononuclear cells, while fraction 0 induced the lowest proliferative response among patients with PCM, in either acute, acute treated, or chronic forms.

Immunization of bovines using a DNA vaccine (pcDNA3.1/MSP1b) prepared from the Jaboticabal strain of Anaplasma marginale.

Anaplasma is a tick-borne ehrlichial pathogen of cattle that causes the disease, anaplasmosis. In the present study, a total of 11 Anaplasma marginale seronegative calves were assigned into two groups: one immunized (G1, n = 6) and one nonimmunized-control (G2, n = 5). Six calves were immunized by using a DNA vaccine containing the gene of a major surface protein, MSP1b, encoded by the plasmid identified as pcDNA3.1/MSP1b. Calves received three intramuscular inoculations of 100 microg of pcDNA3.1/MSP1b at a 20-day interval. The control group received buffer phosphate at the same schedule as the experimental group. The immune response elicited by immunization with pcDNA3.1/MSP1b was evaluated in mice and calves. Twenty days following initial immunization, specific serum antibody from four BALB/c mice bound MSP1b in immunoblots. Sixty days after the last immunization, all calves were challenged with cryopreserved A. marginale at a dose of 10(4) parasites/mL/animal by intravenous injection. Results of packed cell volume (PCV) and detection of infected erythrocytes in all experimental groups revealed that the decrease of PCV and detection of infected erythrocytes occurred at 28 to 42 days after challenge. Mean temperature values did not increase over 39.85 degrees C. Antibodies developed by immunized bovines from G2 were detected 14 days after challenge. MSP1b was characterized during the immunization period and MSP2 was the most predominant polypeptide at the challenge period. DNA of A. marginale was detected in all groups just after challenge by nested PCR assay. It can be concluded that all immunized bovines were partially protected against homologous challenge.

Evidence for the participation of proteinases released by Candida albicans in the early killing of peritoneal macrophages in vitro.

1. We have investigated the possibility that proteinases released by Candida albicans participate in the early killing of three types of mice peritoneal macrophage (resident, thioglycollate-elicited, or Con A-activated) in vitro. 2. Phagocytic assays were performed by incubation of macrophages and C. albicans together at a 1:10 ratio for 30 min at 37 degrees C in RPMI medium buffered to pH 7.0 with 12 mM Hepes without serum. With no albumin added to the medium, the macrophages were 85% to 100% damaged and unviable, to a greater extent than expected from the proportion of phagocytic cells containing germ tubes. When 10 mg/ml of albumin was added to the medium, however, 90% of the macrophages remained viable for the 2 hours of the phagocytic assay, suggesting that albumin may have acted as a substrate for or inhibitor of proteinases released by C. albicans, thereby protecting the macrophage from the proteolytic action of the proteinases. 3. The phagocytosis of IgG-coated erythrocytes was reduced to 43% when IgG was preincubated with the supernatant from C. albicans cultures, but the addition of 10 mg/ml albumin or of 5 micrograms/ml pepstatin (an inhibitor of C. albicans acid proteinases) to the same supernatants prevented the effect on phagocytosis of IgG-coated erythrocytes. 4. These results suggest that proteinases released from C. albicans are involved in the early killing of macrophages.

Congenital cytomegalovirus infection in a neonatal intensive care unit in brazil evaluated by PCR and association with perinatal aspects.

Cytomegalovirus (CMV) infection is the most common congenital infection, affecting 0.4% to 2.3% newborns. Most of them are asymptomatic at birth, but later 10% develop handicaps, mainly neurological disturbances. Our aim was to determine the prevalence of CMV shed in urine of newborns from a neonatal intensive care unit using the polymerase chain reaction (PCR) and correlate positive cases to some perinatal aspects. Urine samples obtained at first week of life were processed according to a PCR protocol. Perinatal data were collected retrospectively from medical records. Twenty of the 292 cases (6.8%) were CMV-DNA positive. There was no statistical difference between newborns with and without CMV congenital infection concerning birth weight (p=0.11), gestational age (p=0.11), Apgar scores in the first and fifth minutes of life (p=0.99 and 0. 16), mother's age (p=0.67) and gestational history. Moreover, CMV congenital infection was neither related to gender (p=0.55) nor to low weight (<2,500 g) at birth (p=0.13). This high prevalence of CMV congenital infection (6.8%) could be due to the high sensitivity of PCR technique, the low socioeconomic level of studied population or the severe clinical status of these newborns.

Catechin attenuates behavioral neurotoxicity induced by 6-OHDA in rats.

This study was designed to investigate the beneficial effect of catechin in a model of Parkinson's disease. Unilateral, intrastriatal 6-hydroxydopamine (6-OHDA)-lesioned rats were pretreated with catechin (10 and 30 mg/kg) by intraperitoneal (i.p.) injection 2h before surgery and for 14 days afterwards. After treatments, apomorphine-induced rotations, locomotor activity, working memory and early and late aversive memories were evaluated. The mesencephalon was used to determine the levels of monoamines and measurement of glutathione (GSH). Immunohistochemical staining was also used to evaluate the expression of tyrosine hydroxylase (TH) in mesencephalic and striatal tissues. Catechin administration attenuated the increase in rotational behavior and the decrease in locomotor activity observed in lesioned rats. Although catechin did not rescue the impairment of late aversive memory, it protected the animals against 6-OHDA-induced working memory deficits. Furthermore, catechin treatment restored GSH levels, and significantly increased dopamine and DOPAC content, and TH-immunoreactivity in 6-OHDA-lesioned rats. Catechin protected 6-OHDA-lesioned rats due to its antioxidant action, indicating that it could be useful as an adjunctive therapy for the treatment of Parkinson's disease.

Efficacy of the Bm86 antigen against immature instars and adults of the dog tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae).

The Bm86 antigen has been used to control ticks of the Boophilus genera in integrated programs that also include the use of acaricides. Because of recent phylogenetic studies have lead to the inclusion of all Boophilus species within the Rhipicephalus genera, we aimed to investigate the efficacy of the Bm86 antigen on the biotic potential of Rhipicephalus sanguineus. Domestic dogs were vaccinated with Bm86 and challenged with the three instars of R. sanguineus. Male and female mongrel dogs were divided into two groups of four animals each, comprising non-vaccinated and vaccinated animals. Immunized dogs were given two doses of an experimental formulation containing 50mug of recombinant Bm86, at 21 days interval while the other group was given placebo, consisting of the same preparation without Bm86. Each dog was challenged 21 days after the last dose with 250 larvae, 100 nymphs and 55 adults (25 females and 30 males) released inside feeding chambers (one per instar) glued to their shaved flank. The effect of the vaccination was evaluated by determining biological parameters of ticks including the yield rates of larvae, nymphs and adult females. Adult females engorged weight, egg mass weight, efficiency rate of conversion to eggs (ERCE) and hatchability. In addition, sera were collected from dogs at 0, 21, 36, 45 and 75 days after the vaccination and used for the detection of specific antibodies by ELISA. Collection rates of larvae, nymphs and adult females fed on vaccinated dogs were significantly (p<0.05) reduced by 38%, 29% and 31%, respectively, as compared with non-vaccinated controls. Significant reductions were also observed in weight of engorged females and egg mass, in ERCE, but not in the hatch rate of ticks fed on immunized dogs. ELISA data revealed a marked and significant increase in optical densities of sera from vaccinated animals after the second dose of Bm86. We concluded that the Bm86 antigen used as a vaccine for dogs reduced the viability and biotic potential of the R. sanguineus.

Topical anti-inflammatory potential of Physalin E from Physalis angulata on experimental dermatitis in mice.

The anti-inflammatory effect of physalin E, a seco-steroid isolated from Physalis angulata L. was evaluated on acute and chronic models of dermatitis induced by 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and oxazolone, respectively, in mouse ear. The changes in ear edema/thickness, production of pro-inflammatory cytokines (TNF-alpha and IFN-gamma), myeloperoxidase (MPO) activity, and histological and immunohistochemical findings were analysed, as indicators of dermal inflammation. Similar to dexamethasone, topically applied Physalin E (0.125; 0.25 and 0.5 mg/ear) potently inhibited the TPA and oxazolone-induced dermatitis, leading to substantial reductions in ear edema/thickness, pro-inflammatory cytokines, and MPO activity. These effects were reversed by mifepristone, a steroid antagonist and confirmed by immunohistochemical and histopathological analysis. The data suggest that physalin E may be a potent and topically effective anti-inflammatory agent useful to treat the acute and chronic skin inflammatory conditions.