Effects of lycopene from guava (Psidium guajava L.) derived products on breast cancer cells.

Polyphenols, condensed tannins, total flavonoids, total carotenoids, lycopene and ascorbic acid were determined besides verifying antioxidant capacity of peel, pulp and desserts (with and without sugar) of red guava (Psidium guajava L.) as well as the effects of lycopene on cytotoxicity, cell cycle and apoptosis on breast cancer cell line MCF-7. Guava peel contains 90% of the total ascorbic acid and heat treatment does not modify bioactive compounds content and antioxidant capacity. Sugar addition decreased guava pulp functional capacity. After heat treatment, lycopene content was stable, but sugar addition reduced its concentration by 57%. Lycopene (10 µM) extracted from guava and standard presented the same cytotoxic effect on MCF-7 cells. Lycopene influenced over G2-M transition check-point of the cell cycle and increased apoptotic cells percentages compared to untreated cells. The consumption of in natura guava, especially with peel can be considered an important source of bioactive compounds.

Could implant position influence the peri-implant parameters in edentulous mandibles of diabetics rehabilitated with overdentures? A split-mouth randomized study.

This randomized controlled trial assessed the impact of crestal level position of implants installed in type 2 diabetes mellitus (T2DM) patients rehabilitated with overdentures. Twenty-two mandibular edentulous T2DM patients were submitted to implant placement for retention of an overdenture. By means of a split-mouth design, two implants were installed: one at supracrestal level (SL) and one at crestal level (CL). Clinical, immunoenzymatic and tomographic analyses were performed at prosthesis placement (baseline) and after 6, 12 and 24 months following implant loading. Increased peri-implant probing depths were detected in CL implants when compared with SL implants at all time-points (baseline P=0.047; 6 months P=0.014; 12 months P=0.027; 24 months P=0.036). Indeed, augmented clinical attachment levels were also detected in CL implants when compared with SL implants at all time-points (baseline P=003; 6 months P=0.045; 12 months P=0.029; 24 months P=0.026). CL implants demonstrated increased amounts of interleukin-6 (IL-6) at 6 months (P=0.043) and higher IL-17 (P=0.021), IL-21 (P=0.034) and tumour necrosis factor alpha (TNF-α) concentrations (P=0.030) at 24 months in comparison with SL implants. CL group revealed enhanced bone loss from baseline to 6 (P=0.032), 12 (P=0.043) and 24 months (P=0.028) when compared with SL. In conclusion, this study showed that implants placed supracrestally in T2DM patients rehabilitated with overdentures demonstrated lower bone loss and better clinical parameters with beneficial modulation of peri-implant immunoinflammatory biomarkers when compared with implants positioned at crestal level.

Optimizing the storage of chemotherapeutics for ophthalmic oncology: stability of topotecan solution for intravitreal injection.

BACKGROUND: . Intravitreal administration of topotecan shows activity against tumor vitreous seeding in the conservative treatment of retinoblastoma, a malignant tumor originated in the retina of small children. Adequate storage of the intravitreal topotecan solution would allow immediate availability for patients at health care institutions. The goal of the work was to address the stability of the intravitreal topotecan formulation upon reconstitution. MATERIALS AND METHODS: . Intravitreal topotecan solutions were reconstituted (at a concentration of 0.2 mg topotecan in 1 mL saline solution vehicle, aliquoted in 1 mL plastic syringes) and stored either frozen or at room temperature for different times. Topotecan content was analyzed at time zero and at different conditions using a high performance liquid chromatography method to quantify topotecan lactone (active) and to detect its pH-dependent hydrolysis product, the open carboxylate. RESULTS: . We found that intravitreal topotecan syringes remained stable at room temperature at least for 24 h, at least for 167 days upon stored frozen at -20°C, and up to 8 h after thawing at day 6. The degradation carboxylate product did not appear in the analyzed thawed samples during the whole study. CONCLUSIONS: . This study confirms the stability of frozen intravitreal topotecan syringes and will help optimize the use of this chemotherapy modality at institutions with low resources. Storage of aliquots will also help reduce personnel exposure to chemotherapy at hospital pharmacies.

The impact of implant abutment surface treatment with TiO2 on peri-implant levels of angiogenesis and bone-related markers: a randomized clinical trial.

The goal of this randomized, blinded, split-mouth controlled clinical trial was to assess the influence of abutment surface treatment on tissue healing. Fifteen patients received two implants distributed randomly to two groups: test (TiO2 abutment surface), control (standard abutment surface). Levels of epidermal growth factor (EGF), bone morphogenetic protein 9 (BMP-9), endothelin 1 (ET-1), fibroblast growth factor (FGF), placental growth factor (PlGF), and vascular endothelial growth factor (VEGF) were quantified in the peri-implant fluid after 3, 14, 30, and 60 days. Inter-group comparisons indicated higher levels of EGF, BMP-9, ET-1, FGF, and PlGF in the test group after 30days (P<0.05). PlGF levels were also higher in the test group after 60 days. In the test group, intra-group analysis revealed different levels of ET-1 and FGF between days 3 and 30, and days 3 and 60 (P<0.05); furthermore, VEGF levels were significantly higher on day 60 than on day 3 (P <0.05). In the control group, intra-group analysis demonstrated significantly different levels of ET-1, FGF, and PlGF between days 3 and 60 and of PlGF between days 14 and 60 (P<0.05). In conclusion, abutment surfaces treated with TiO2 influenced the levels of angiogenesis and bone-related markers.

Effect of curcumin on bone tissue in the diabetic rat: repair of peri-implant and critical-sized defects.

This study determined the effect of curcumin on bone healing in animals with diabetes mellitus (DM). One hundred rats were divided into five groups: DM+PLAC, DM+CURC, DM+INS, DM+CURC+INS, and non-DM (CURC, curcumin; PLAC, placebo; INS, insulin). Critical calvarial defects were created and titanium implants were inserted into the tibiae. Calvarial defects were analyzed histometrically, and BMP-2, OPN, OPG, RANKL, Runx2, Osx, ß-catenin, Lrp-5, and Dkk1 mRNA levels were quantified by PCR. The implants were removed for a torque evaluation, the peri-implant tissue was collected for mRNA quantification of the same bone-related markers, and the tibiae were submitted to micro-computed tomography. The DM+CURC+INS and non-DM groups exhibited greater closure of the calvaria when compared to the DM+PLAC group (P<0.05). Increased retention of implants was observed in the DM+CURC, DM+CURC+INS, and non-DM groups when compared to the DM+PLAC group (P<0.05). CURC improved bone volume and increased bone-implant contact when compared to DM+PLAC (P<0.05). In calvarial samples, CURC favourably modulated RANKL/OPG and Dkk1 and improved ß-catenin levels when compared to DM+PLAC (P<0.05). In peri-implant samples, Dkk1 and RANKL/OPG were down-regulated and BMP-2 up-regulated by CURC when compared to DM+PLAC (P<0.05). CURC reverses the harmful effects of DM in bone healing, contributing to the modulation of bone-related markers.

The impact of different torques for the insertion of immediately loaded implants on the peri-implant levels of angiogenesis- and bone-related markers.

The aim of this split-mouth, randomized, double-blind, controlled clinical trial was to evaluate the influence of different insertion torque values for dental implants on bone- and angiogenesis-related marker profiles. Eighteen edentulous patients received dental implants and fixed complete-arch mandibular prostheses. The implants (n=36) were assigned randomly to two groups: reduced torque (n=18), with insertion torque <30Ncm; and conventional torque (n=18), with insertion torque ≥30Ncm. Levels of vascular endothelial growth factor (VEGF), placental growth factor, bone morphogenetic protein 9 (BMP-9), periostin, osteoprotegerin (OPG), and tartrate-resistant acid phosphatase (TRAP) in the peri-implant fluid were quantified at 7, 14, 30, and 120days after implant placement. Inter-group comparisons showed that VEGF and OPG levels were higher in the low-level torque group than in the conventional torque group on days 7 and 30, respectively (P<0.05). BMP-9 and periostin levels were higher in the conventional group than in the low-level torque group on day 120, and TRAP was up-regulated around implants inserted with conventional torque when compared to those inserted with lower-level torque at all time points evaluated (P<0.05). In conclusion, the use of different levels of torque for implantation of immediately loaded implants significantly influenced the levels of bone- and angiogenesis-related markers during early peri-implant repair.

Impact of resveratrol on bone repair in rats exposed to cigarette smoke inhalation: histomorphometric and bone-related gene expression analysis.

This study investigated the effect of resveratrol on bone healing and its influence on the gene expression of bone-related markers in rats exposed to cigarette smoke. Two calvarial defects were created in each of 60 rats, which were assigned equally (n=20) to three groups: (1) resveratrol (10mg/kg)+smoke exposure (SMK+RESV); (2) placebo+smoke exposure (SMK+PLA); or (3) placebo+no smoke exposure (NS+PLA). Substances were administered daily for 30days following surgery. Smoke inhalation was started 7days before surgery and continued for 30days after surgery. One defect was processed for histomorphometric analysis and the other was used for mRNA quantification of bone-related gene expression by qPCR. The remaining defect was smaller in the SMK+RESV (2.27±0.61mm, P=0.0003) and NS+PLA (2.17±0.74mm, P=0.0005) groups than in the SMK+PLA group (3.12±0.47mm). Higher levels of Runx2 were observed in the NS+PLA group than in the smoke exposure groups (vs. SMK+PLA, P=0002; vs. SMK+RESV, P=0.052); levels of Lrp-5 were also higher in the no smoke exposure group (vs. SMK+RESV, P=0.009; vs. SMK+PLA, P=0.003). Resveratrol therapy decreased RANKL/OPG expression when compared to placebo (SMK+RESV vs. SMK+PLA, P=0.017). Dkk1 levels were decreased in the SMK+RESV group when compared to the SMK+PLA (P=0.006) and NS+PLA groups (P=0.005). In conclusion, resveratrol optimizes the repair of critical-sized bone defects, up-regulating the gene expression of important bone remodelling markers in rats exposed to cigarette smoke inhalation.

Systemic treatment with resveratrol and/or curcumin reduces the progression of experimental periodontitis in rats.

BACKGROUND AND OBJECTIVE: Periodontitis is a chronic inflammatory disease of periodontal tissues that leads to the destruction of bone and other connective tissues. Resveratrol and curcumin are plant-derived substances with biological properties that may have immunomodulatory properties. This study investigated the effect of continuous administration of resveratrol and curcumin and the association of resveratrol and curcumin on the progression of experimental periodontitis in rats. MATERIAL AND METHODS: Forty Wistar rats were assigned randomly to the following groups: group 1, experimental periodontitis + placebo (PL) (n = 10); group 2, experimental periodontitis + resveratrol (RSV) (n = 10); group 3, experimental periodontitis + curcumin (C) (n = 10); and group 4, experimental periodontitis + resveratrol + curcumin (COMBI) (n = 10). Periodontitis was induced in rats by tying a silk suture, as a ligature, around one of the first molars. Daily administration of the placebo solution, 10 mg/kg of resveratrol, 100 mg/kg of curcumin or 10 mg/kg of resveratrol plus 100 mg/kg of curcumin was carried out from day 0 to day 30. At the end of the relevant experimental periods, rats were killed and the specimens obtained were processed for morphometric analysis of bone loss. Gingival tissues surrounding the first molar were collected for quantification of interleukin (IL)-1ß, IL-4, interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) using a Luminex/MAGPIX assay. RESULTS: Intergroup comparisons of the morphometric outcomes revealed higher bone-loss values in the PL group (p < 0.05) when compared with RSV, C and COMBI groups. There was no difference in bone-loss values among RSV, C and COMBI groups (p > 0.05). The immunoenzymatic assay of the gingival tissue showed a lower concentration of IL-1ß in the COMBI group in comparison with the PL group (p < 0.05). Higher values of IL-4 were demonstrated in groups RSV, C and COMBI in comparison with the PL group (p < 0.05). Only RSV caused a reduction in the levels of IFN-γ (p < 0.05). There was no difference in the concentration of TNF-α amongst the four groups (p > 0.05). CONCLUSION: Resveratrol and curcumin are capable of reducing alveolar bone loss in an animal model of periodontitis. This occurred when these agents were added singly or in combination with one another, but there did not appear to be either synergistic or additive effects.

Alcohol intake may impair bone density and new cementum formation after enamel matrix derivative treatment: histometric study in rats.

BACKGROUND AND OBJECTIVE: Alcohol intake may interfere with bone metabolism; however, there is a lack of information about the outcomes of regenerative approaches in the presence of alcohol intake. Enamel matrix derivative (EMD) has been used in periodontal regenerative procedures resulting in improvement of clinical parameters. Thus, the aim of this histomorphometric study is to evaluate the healing of periodontal defects after treatment with EMD under the influence of alcohol intake. MATERIAL AND METHODS: Twenty Wistar rats were randomly assigned to two groups: G1 = alcohol intake (n = 10) and G2 = non-exposed to alcohol intake (n = 10). Thirty days after initiation of alcohol intake, fenestration defects were created at the buccal aspect of the first mandibular molar of all animals from both groups. After the surgeries, the defects of each animal were randomly assigned to two subgroups: non-treated control and treated with EMD. The animals were killed 21 d later. RESULTS: G1 showed less defect fill for non-treated controls. Bone density (BD) and new cementum formation were lower for G1 when compared to G2, for EMD-treated and non-treated sites. EMD treatment resulted in greater BD and new cementum formation in both groups and defect fill was not significantly different between groups in the EMD-treated sites. The number of tartrate-resistant acid phosphatase-positive osteoclasts was significantly higher in G1 when compared to G2 and in EMD-treated sites of both groups. CONCLUSION: Alcohol intake may produce a significant detrimental effect on BD and new cementum formation, even in sites treated with EMD. A limited positive effect may be expected after EMD treatment under this condition.

Cigarette smoke inhalation increases the alveolar bone loss caused by primary occlusal trauma in a rat model.

BACKGROUND AND OBJECTIVE: Occlusal trauma (OT) and smoking are both factors that alter alveolar bone metabolism and therefore could synergistically act on alveolar bone loss. The aim of this experimental study was to evaluate the influence of short-term cigarette smoke inhalation (CSI) on inter-radicular alveolar bone loss promoted by primary OT in a rat model. MATERIAL AND METHODS: Forty-eight animals were randomly assigned to one of three groups based on treatment type: OT + CSI (n = 16), animals were exposed to CSI three times per day, for 8 min per exposure, and they concomitantly received unilateral vertical augmentation creating an occlusal interference inducing experimental OT; OT (n = 16), animals received only unilateral vertical augmentation; negative control (NC; n = 16), animals maintained for equal periods to achieve periodontal baseline values of periodontal ligament dimension. Each group was divided into two subgroups (n = 8) based on treatment length: 7 or 14 d. RESULTS: After 7 d, the OT + CSI group exhibited significantly higher bone loss compared to the NC group (p = 0.0022). After 14 d, the OT (p < 0.0001) and OT + CSI (p < 0.0001) groups presented significantly higher bone loss compared to the NC group, and OT + CSI resulted in significantly higher bone loss than OT alone (p = 0.0241). The number of tartrate-resistant acid phosphatase-positive cells on the linear surface of the bone crest after 7 d was significantly higher in the OT + CSI group as compared to the NC and OT groups (p < 0.0001 and p = 0.0045, respectively) and remained significantly higher in the OT + CSI group after 14 d, compared to the OT group (p < 0.0001). CONCLUSION: Short-term CSI increases early bone loss in association with OT after 7 d, and this worsens in severity after 14 d of exposure.

Antimicrobial susceptibility of coagulase-negative Staphylococci isolated from mastitic cattle in Brazil / Susceptibilidade antimicrobiana de cepas de Staphylococci coagulase-negativa isoladas de leite de bovinos com mastite no Brasil

Um total de 109 cepas de Staphylococci coagulase-negativa foi isolado de leite de vacas com mastite clínica e subclínica, em 35 fazendas, situadas em nove estados brasileiros, no período de fevereiro a maio de 2005. Os isolados foram investigados em relação a susceptibilidade in vitro a diversos agentes antimicrobianos. A resistência à penicilina foi a observação mais freqüente (93,5 por cento), seguida por sulfonamida (88,9 por cento), novobiocina (88,6 por cento) e ampicilina (85,3 por cento). Todas as cepas examinadas mostraram resistência a pelo menos uma das drogas antimicrobianas testadas. Cepas apresentando resistência múltipla foram extremamente comuns, com 10,0 por cento dos microrganismos isolados apresentando resistência a todas as drogas antimicrobianas. Os resultados obtidos indicaram que as cepas de Staphylococci coagulase-negativas, isoladas no Brasil, apresentaram um alto grau de resistência a antimicrobianos. Estes resultados são, provavelmente, uma conseqüência da pressão devida ao uso intensivo de drogas antimicrobianas.

O-serogroups, eae gene and EAF plasmid in Escherichia coli isolates from cases of bovine mastitis in Brazil.

Mastitis has been recognized for some time as the most costly disease in dairy herds. From March 1997 to August 1998, 2144 samples of bovine mastitic milk were collected, from which 182 Escherichia coli isolates were made, and from which 141 isolates had the somatic antigen (serogroup) determined. Twelve different serogroups were isolated from mastitic milk, and among them were O26, O55, O111 and O119, all of them classic enteropathogenic E. coli (EPEC) serogroups. These represented 40.0% of the isolates. The 20 of 57 isolates tested had plasmids and in dot blot hybridization, nine isolates were positive for an EaeA probe and an EPEC adherence factor (EAF) probe while two isolates were negative for EaeA probe but positive for the EAF probe. The nine isolates were characterized as attaching and effacing (A/E) E. coli (AEEC) isolates.

Early treatment of an intruded primary tooth: a case report.

The report is about a case of total intrusion of a deciduous central incisor involving a male patient of 14 months of age. Repositioning of the intruded tooth was performed 20 minutes after the trauma, followed by placement with sutures. Clinical and radiographic monitoring performed after 1, 2, 4, 9, and 13 months showed normal characteristics. We emphasize the development of preventive programs promoting parental awareness of the importance of looking for prompt care, resulting in a better prognosis.

Presence of a member of the mitochondrial carrier family in hydrogenosomes: conservation of membrane-targeting pathways between hydrogenosomes and mitochondria.

A number of microaerophilic eukaryotes lack mitochondria but possess another organelle involved in energy metabolism, the hydrogenosome. Limited phylogenetic analyses of nuclear genes support a common origin for these two organelles. We have identified a protein of the mitochondrial carrier family in the hydrogenosome of Trichomonas vaginalis and have shown that this protein, Hmp31, is phylogenetically related to the mitochondrial ADP-ATP carrier (AAC). We demonstrate that the hydrogenosomal AAC can be targeted to the inner membrane of mitochondria isolated from Saccharomyces cerevisiae through the Tim9-Tim10 import pathway used for the assembly of mitochondrial carrier proteins. Conversely, yeast mitochondrial AAC can be targeted into the membranes of hydrogenosomes. The hydrogenosomal AAC contains a cleavable, N-terminal presequence; however, this sequence is not necessary for targeting the protein to the organelle. These data indicate that the membrane-targeting signal(s) for hydrogenosomal AAC is internal, similar to that found for mitochondrial carrier proteins. Our findings indicate that the membrane carriers and membrane protein-targeting machinery of hydrogenosomes and mitochondria have a common evolutionary origin. Together, they provide strong evidence that a single endosymbiont evolved into a progenitor organelle in early eukaryotic cells that ultimately give rise to these two distinct organelles and support the hydrogen hypothesis for the origin of the eukaryotic cell.

Cervico-vaginal Chlamydia trachomatis infection in pregnant adolescent and adult women. A morphologic and immunofluorescent study.

We studied the incidence of cervical Chlamydia trachomatis (CT) in 65 adolescents aged between 14 and 19 years and 65 adults aged 20 years or more. All subject were more than three months advanced in a normal pregnancy. Two samples for cytology were obtained from each patient, one ecto- and endocervical, with Ayre's spatula and cytobrush, and one only endocervical with cytobrush. The first one was examined after Papanicolaou staining and the second one was examined with the easily performed immunofluorescence reaction to CT (IF)--Microtrak, SYVA. Twenty seven adolescent patients (41.5%) and fourteen adults (21.5%) had a positive IF test for CT; the rate in adolescents was significantly higher (P < 0.01) than in adults. The Papanicolaou (Pap) stained slides, examined blind for evidence of CT infection showed a sensitivity of 70.7%, a specificity of 95.5% and positive and negative predictive values of 87.8% and 87.6% respectively. Seven adolescents showed cytological signs of Papillomavirus (HPV) infection, and six of them were also CT positive; five had signs of HPV infection and 4 of them were CT positive.