Genotoxic and mutagenic effects of methyl ether dillapiole on the development of Aedes aegypti (Diptera: Culicidae).

Dillapiole, extracted from Piper aduncum essential oil and its derivatives, has been shown to be a potential alternative to the control of Aedes aegypti, which has become resistant to synthetic insecticides. Methyl ether dillapiole (MED) and temephos (TM) were compared to complement the data on the genotoxicity and developmental changes of Ae. aegypti. Over four generations (G1 -G4 ), third stage larvae were treated with MED at 60, 80 and 100 µg/mL and TM at 0.002, 0.005 and 0.007 µg/mL for 4 h. Adult females were separated to estimate oviposition and hatching rates, and total egg length. Over the four generations, a significant reduction was recorded in oviposition and hatching rates, and in mean egg length (Tukey, P < 0.05), compared with the negative control (NC). Cytological slide preparations were done from adult oocytes and larval neuroblasts. The cumulative effects of genotoxic (bridges, budding and nuclear fragmentation) and mutagenic (micronucleus and chromosomal breakage) damage was observed in the neuroblasts and oocytes of exposed mosquitoes. Developmental changes and damage to the genome of MED-treated Ae. aegypti were greater than those caused by TM. Further studies should focus on understanding the effects of the MED molecule on Ae. aegypti.

Effects of canola oil and antioxidants on performance, serum parameters, carcass traits, and rumen fermentation patterns of Nellore cattle.

Several nutritional strategies have been used in beef cattle production in order to increase animal performance and profitability. However, in the past two decades, the increase of consumer preference for functional foods has driven the investigation for improving food via adding functional substances to animal diets. We evaluated the effect of canola oil supplementation associated with vitamin E and selenium on performance, rumen metabolism, carcass traits, meat tenderness, and serum, liver, and meat status of antioxidants in finishing Nellore males. Animals were fed for 106 days in a feedlot and were randomly distributed in a 2 × 2 factorial arrangement: two levels of oil in the diet (no inclusion and 3% canola oil, defined as diet without oil inclusion (NO) and effect of oil (OIL), respectively) and two levels of antioxidants in the diet (no inclusion and 2.5 mg of Se/kg of DM + 500 UI of vitamin E/kg of DM, defined as diet without antioxidant inclusion (NA) and effect of the antioxidants (ANT), respectively). DM intake (kg/day) was evaluated daily; performance and serum were analysed at the beginning of the feedlot and every 28 days. Animals were slaughtered and hot carcass weight (kg) was recorded; ruminal fluid and liver samples were collected. At 24 h postmortem, carcass pH was recorded and the Longissimus thoracis was sampled. There was no significant effect of the OIL*ANT interaction (P > 0.05) for any trait evaluated. Bulls fed OIL presented greater final BW (P < 0.01), average daily gain (kg/day; P < 0.01), feed efficiency (P < 0.01), rump fat thickness (P8RF; P < 0.05), and greater tenderness; the ANT diet increased P8RF (P < 0.05). The levels of selenium and vitamin E in serum, liver, and meat were increased (P < 0.01) with the inclusion of ANT. ANT did not change triiodothyronine (T3, ng/mL) and thyroxine (T4, µg/gL) serum concentrations but decreased serum glucose levels. The treatments did not affect (P > 0.05) ruminal parameters or the protozoa population. Our results showed that the inclusion of 3% canola oil in the diet DM increased performance, feed efficiency, carcass fat deposition, and tenderness, with no effect on rumen fermentation and protozoa population of Nellore cattle in a feedlot system. The inclusion of ANT in the cattle diet did not affect performance or rumen parameters. However, the levels of ANT were increased in the serum, liver, and meat, enriching the final product with these compounds.

Biomechanical analysis in mandibular advancement and occlusal plane rotation with finite element analysis.

We used finite element analysis to assess stress on the cortical bone and plate fixation system, as well as mandibular resistance after sagittal split ramus osteotomy with different mandibular advancements and rotations of the occlusal plane. Three-dimensional mandibular models were obtained, and 6mm and 12mm advancements were planned associated with linear, clockwise, and counter-clockwise rotation of the angle of the occlusal plane. Each model was then fixed with one or two 2.0mm system plates and secured with four monocortical screws. A total of 12 models were built and subjected to a vertical load in the lower central incisor ranging from 50N to 500N in 50N increments. Results showed that the 12mm advancement was associated with higher stress on the bone and plate surface. Additionally, the models fixed with two plates exhibited lower plate stress than those fixed with a single plate. Counter-clockwise rotation of the angle of the mandibular plane in the 6mm advancement caused more plate stress, which did not occur in the 12mm advancement. This analysis has shown that change in the occlusal plane in large mandibular advancements does not act as an additional stress factor. These findings can help to better understand the tensions on bone and plate surfaces in patients who need large mandibular advancements that are associated with a change in the occlusal plane, and will aid better surgical planning.

Guanosine enhances glutamate uptake and oxidation, preventing oxidative stress in mouse hippocampal slices submitted to high glutamate levels.

Glutamate (Glu) is the main mammalian brain neurotransmitter. Concerning the glutamatergic neurotransmission, excessive levels of glutamate in the synaptic cleft are extremally harmful. This phenomenon, named as excitotoxicity is involved in various acute and chronic brain diseases. Guanosine (GUO), an endogenous guanine nucleoside, possesses neuroprotective effects in several experimental models of glutamatergic excitotoxicity, an effect accompanied by an increase in astrocytic glutamate uptake. Therefore, the objective of this study was to investigate the involvement of an additional putative parameter, glutamate oxidation to CO2, involved in ex-vivo GUO neuroprotective effects in mouse hippocampal slices submitted to glutamatergic excitotoxicity. Mice were sacrificed by decapitation, the hippocampi were removed and sliced. The slices were incubated for various times and concentrations of Glu and GUO. First, the concentration of Glu that produced an increase in L-[14C(U)]-Glu oxidation to CO2 without cell injury was determined at different time points (between 0 and 90 min); 1000 µM Glu increased Glu oxidation between 30 and 60 min of incubation without cell injury. Under these conditions (Glu concentration and incubation time), 100 µM GUO increased Glu oxidation (35%). Additionally, 100 µM GUO increased L-[3,4-3H]-glutamate uptake (45%) in slices incubated with 1000 µM Glu (0-30 min). Furthermore, 1000  µM Glu increased reactive species levels, SOD activity, and decreased GPx activity, and GSH content after 30 and 60 min; 100 µM GUO prevented these effects. This is the first study demonstrating that GUO simultaneously promoted an increase in the uptake and utilization of Glu in excitotoxicity-like conditions preventing redox imbalance.

The signal transducer and activator of transcription 6 (STAT-6) mediates Th2 inflammation and tissue damage in a murine model of peanut-induced food allergy.

INTRODUCTION: Food allergies are inflammatory conditions mediated by Th2 and probably STAT-6 dependent immune responses. OBJECTIVE AND DESIGN: Here we investigated the role of Signal Transducer and Activator of Transcription 6 (STAT-6) in development of inflammation in peanut allergy. METHODS: To induce food allergy, wild-type (WT) and mice deficient for STAT-6 (Stat6-/-) were sensitized with peanut proteins and challenged with peanut seeds. RESULTS: WT animals lost weight and refused the peanut diet, in contrast to Stat6-/- mice, which had a better maintenance of body weight and more regular seeds' consumption. The augmented peanut-specific IgG, IgG1 and IgE in the allergic WT was abolished in Stat6-/- animals that also presented increased IgG2a. There was an overall reduction in the gut mediators in the absence of STAT-6, including those related to inflammatory and Th2 responses, in contrast to a rising counter regulatory and Th1 reaction in Stat-6-/- mice. These animals had IFN-γ and IL-10 similar to WT after the four-week challenge. Most interestingly, Stat-6-/- mice had no intestinal damage, in contrast to WT animals, which had inflammatory infiltrate, tissue destruction, epithelial exulceration, edema, congestion and loss of villous architecture in the small gut segments. CONCLUSIONS: STAT-6 plays an important role in the establishment of the Th2 inflammatory responses and intestinal damage in peanut allergy.

Biomechanical simulation of temporomandibular joint replacement (TMJR) devices: a scoping review of the finite element method.

The aim of this study was to perform a literature review on the use of finite element modeling (FEM) for the evaluation of the biomechanical behavior of temporomandibular joint replacement (TMJR) devices. An electronic search of online medical and scientific literature database was conducted using selected search terms. The search identified 307 studies, of which 19 were considered relevant to this study. Of the 19 selected studies, 10 (52.6%) investigated the influence of geometry and fixation methods, while two (10.5%) evaluated the behavior of artificial condyle-fossa structures. The TMJR devices assessed in these studies included TMJ Inc. (aka Christensen; 63.2%), Zimmer Biomet (15.7%), Stryker (10.5%), and a theoretical intramedullary condylar component (5.3%); 26.3% of the studies evaluated custom TMJR devices. Such studies provided important data on the distribution of strain and stress through TMJR structural components and surrounding bone by using different software systems and methods. The mean stress values were lower on a custom TMJR condyle-ramus component and the supporting bone than on the stock device. FEM proved to be an accurate and valuable biomechanical simulation tool for studying the current TMJR devices and should be considered a useful tool for the improvement and development of future joint replacement devices.

Guanosine Exerts Neuroprotective Effect in an Experimental Model of Acute Ammonia Intoxication.

The nucleoside guanosine (GUO) increases glutamate uptake by astrocytes and acts as antioxidant, thereby providing neuroprotection against glutamatergic excitotoxicity, as we have recently demonstrated in an animal model of chronic hepatic encephalopathy. Here, we investigated the neuroprotective effect of GUO in an acute ammonia intoxication model. Adult male Wistar rats received an intraperitoneal (i.p.) injection of vehicle or GUO 60 mg/kg, followed 20 min later by an i.p. injection of vehicle or 550 mg/kg of ammonium acetate. Afterwards, animals were observed for 45 min, being evaluated as normal, coma (i.e., absence of corneal reflex), or death status. In a second cohort of rats, video-electroencephalogram (EEG) recordings were performed. In a third cohort of rats, the following were measured: (i) plasma levels of glucose, transaminases, and urea; (ii) cerebrospinal fluid (CSF) levels of ammonia, glutamine, glutamate, and alanine; (iii) glutamate uptake in brain slices; and (iv) brain redox status and glutamine synthetase activity in cerebral cortex. GUO drastically reduced the lethality rate and the duration of coma. Animals treated with GUO had improved EEG traces, decreased CSF levels of glutamate and alanine, lowered oxidative stress in the cerebral cortex, and increased glutamate uptake by astrocytes in brain slices compared with animals that received vehicle prior to ammonium acetate administration. This study provides new evidence on mechanisms of guanine-derived purines in their potential modulation of glutamatergic system, contributing to GUO neuroprotective effects in a rodent model of by acute ammonia intoxication.

Evaluation of patients undergoing placement of zygomatic implants using sinus slot technique.

BACKGROUND: This study aimed to evaluate patients undergoing placement of zygomatic implants by Stella and Warner's technique, considering the survival rate of conventional and zygomatic implants, and assess the health of the maxillary sinuses and the level of patient satisfaction. METHODS: In this retrospective cohort study, 28 patients had received a combination of conventional and zygomatic implants (group I) and 14 were rehabilitated with only conventional implants (group II). RESULTS: The results showed that Stella and Warner's technique, thought to minimize the presence of the implant into the maxillary sinus, improving the emergence of the implant, proved to be effective, allowing a high survival rate of conventional and zygomatic implants (100 %). The follow-up period ranged from a minimum of 15 months to a maximum of 53 months after prosthetic rehabilitation (average of 34 months). No pathological changes were found on the periimplant tissues. Radiographs showed satisfactory bone levels in conventional implants of oral rehabilitation with zygomatic implants and a good positioning of the apex of the zygomatic implants in relation to the zygomatic bone. The tomographic findings revealed no characteristics of sinus disease. There were no cases of obstruction of the maxillary sinus ostium. CONCLUSIONS: The placement of zygomatic implants by Stella and Warner's technique proved to be a predictable technique with high implant survival rate in patients with atrophic maxilla and was not associated with sinus disease in the sample analyzed. However, a long-term follow-up is necessary to confirm the initial findings of this study.

Lophine derivatives as activators in peroxyoxalate chemiluminescence.

Lophine and four of its derivatives were used as activators (ACTs) of the chemiluminescent peroxyoxalate (PO) reaction of bis(2,4,6-trichlorophenyl)oxalate with H2O2, catalysed by imidazole. Kinetic emission assays have shown that with the tested compounds the reaction mechanism, regarding the formation of the high energy intermediate (HEI) of the PO reaction, occurs as previously seen for commonly used ACTs. A bimolecular interaction of the compounds with the HEI leads to chemiexcitation through the chemically initiated electron exchange luminescence (CIEEL) mechanism, as confirmed by a linear free-energy correlation between the relative catalytic rate constants and the oxidation potentials of the compounds. The yields of excited state formation and light emission, in the range of 10(-2)-10(-3) E mol(-1), are comparable to the ones seen with commonly used ACTs. A Hammett plot with ρ = -0.90 indicates the buildup of a partial positive charge on the transition step of the catalytic process, consistent with the formation of a radical cation of the ACT, being an additional validation of the CIEEL mechanism in this system.

In vitro mechanical assessment of 2.0-mm system three-dimensional miniplates in anterior mandibular fractures.

This study constituted a comparative assessment of the mechanical resistance of square and rectangular 2.0-mm system three-dimensional miniplates as compared to the standard configuration using two straight miniplates. 90 polyurethane replica mandibles were used for the mechanical trials. Groups 1, 2, and 3 simulated complete symphyseal fractures characterized by linear separation of the central incisors; groups 4, 5, and 6 simulated parasymphyseal fractures with an oblique configuration. Groups 1 and 4 represented the standard method with two straight miniplates set parallel to one another. Square miniplates were used in groups 2 and 5, and rectangular miniplates in groups 3 and 6. A universal testing machine set to a velocity of 10mm/min and delivering a vertical linear load to the first left molar was used to test each group. Maximum load values and load values with pre-established dislocation of 5mm were obtained and submitted to statistical analysis using a calculated reliability interval of 95%. The mechanical performances of the devices were similar, except in the case of rectangular plates used in the parasymphyseal fractures. The innovative fixation methods used showed significantly better results in the case of symphyseal fractures.

Impact of smoking on inflammation: overview of molecular mechanisms.

BACKGROUND: Inflammation is a critical component of normal tissue repair, as well as being fundamental to the body's defense against infection. Environmental factors, such as smoking, have been reported to modify the host response and hence modify inflammation progression, severity and outcome. Therefore, a comprehensive understanding of the molecular mechanisms by which smoking affects inflammation is vital for preventive and therapeutic strategies on a clinical level. AIM: The purpose of the present article is to review the potential biological mechanisms by which smoking affects inflammation, emphasizing recent developments. RESULTS: Smoking is reported to effect a number of biological mediators of inflammation through its effect on immune-inflammatory cells, leading to an immunosuppressant state. Recent evidence strongly suggests that the molecular mechanisms behind the modulation of inflammation by smoking mainly involve the nuclear factor-kappa B (NF-kB) family, through the activation of both an inhibitor of IkB kinase (IKK)-dependent and -independent pathway. In addition to NF-kB activation, a number of transcriptional factors including GATA, PAX5 and Smad 3/4, have also been implicated. CONCLUSION: Multiple mechanisms may be responsible for the association of smoking and inflammation, and the identification of potential therapeutic targets should guide future research.

Factors involved in the T helper type 1 and type 2 cell commitment and osteoclast regulation in inflammatory apical diseases.

INTRODUCTION: Periapical chronic lesion formation involves activation of the immune response and alveolar bone resorption around the tooth apex. However, the overall roles of T helper type 1 (Th1), Th2, and T-regulatory cell (Treg) responses and osteoclast regulatory factors in periapical cysts and granulomas have not been fully determined. This study aimed to investigate whether different forms of apical periodontitis, namely cysts and granulomas, show different balances of Th1, Th2 regulators, Treg markers, and factors involved in osteoclast chemotaxis and activation. METHODS: Gene expression of these factors was assessed using quantitative real-time polymerase chain reaction, in samples obtained from healthy gingiva (n = 8), periapical granulomas (n = 20), and cysts (n = 10). RESULTS: Periapical cysts exhibited a greater expression of GATA-3, while a greater expression of T-bet, Foxp3, and interleukin-10 (IL-10) was seen in granulomas. The expression of interferon-gamma, IL-4, and transforming growth factor-beta was similar in both lesions. Regarding osteoclastic factors, while the expression of SDF-1alpha/CXCL12 and CCR1 was higher in cysts, the expression of RANKL was significantly higher in granulomas. Both lesions exhibited similar expression of CXCR4, CKbeta8/CCL23, and osteoprotegerin, which were significantly higher than in control. CONCLUSION: Our results showed a predominance of osteoclast activity in granulomas that was correlated with the Th1 response. The concomitant expression of Treg cell markers suggests a possible suppression of the Th1 response in granulomas. On the other hand, in cysts the Th2 activity is augmented. The mechanisms of periradicular lesion development are still not fully understood but the imbalance of immune and osteoclastic cell activity in cysts and granulomas seems to be critically regulated by Treg cells.

Brain plasmin enhances APP alpha-cleavage and Abeta degradation and is reduced in Alzheimer's disease brains.

The proteolytic processing of amyloid precursor protein (APP) has been linked to sphingolipid-cholesterol microdomains (rafts). However, the raft proteases that may be involved in APP cleavage have not yet been identified. In this work we present evidence that the protease plasmin is restricted to rafts of cultured hippocampal neurons. We also show that plasmin increases the processing of human APP preferentially at the alpha-cleavage site, and efficiently degrades secreted amyloidogenic and non-amyloidogenic APP fragments. These results suggest that brain plasmin plays a preventive role in APP amyloidogenesis. Consistently, we show that brain tissue from Alzheimer's disease patients contains reduced levels of plasmin, implying that plasmin downregulation may cause amyloid plaque deposition accompanying sporadic Alzheimer's disease.

Up-regulation of T helper 2 and down-regulation of T helper 1 cytokines during murine retrovirus-induced immunodeficiency syndrome enhances susceptibility of a resistant mouse strain to Leishmania amazonensis.

Resistance to and recovery from leishmania infection is dependent on cell-mediated immunity. C57BL/6 mice are resistant to Leishmania amazonensis (La) infection but susceptible to LP-BM5 murine leukemia virus (MuLV) infection. MuLV infection leads to a state of immunodeficiency characterized by severe compromise of cell-mediated immunity. When infected with La alone, C57BL/6 mice developed a small transient lesion that evolved to spontaneous healing or a lesion with extremely slow growth. Lesions were predominantly comprised of a lympho-macrophagic infiltrate with few parasitized macrophages. When infected with La and, 4 weeks later, with MuLV (La-MuLV), the mice developed a large uncontrolled nonhealing lesion containing vacuolated and heavily parasitized macrophages. In contrast, mice infected with MuLV first and La 4 weeks later (MuLV-La) developed a small but persistent lesion, characterized histologically by a small number of heavily parasitized macrophages and few lymphocytes. Eight weeks after MuLV infection, both had similar immunological profiles with decreased lymphocyte proliferation, diminished production of interferon-gamma, and high production of interleukins 4 and 10. At the time of L. amazonensis infection, La-MuLV animals have a normal T cell function whereas in MuLV-La mice this function is already impaired; this may influence the recruitment of macrophages to the site of leishmania injection.

Selective inability of spleen antigen presenting cells from Leishmania donovani infected hamsters to mediate specific T cell proliferation to parasite antigens.

Golden hamsters (Mesocricetus auratus) infected with Leishmania donovani develop a disease similar to human kala-azar. There is conspicuous hypergammaglobulinaemia and their T cells do not respond to stimulation by parasite antigens. The impairment of the cellular immune response seems to be restricted to parasite antigens since infected animals are able to develop a T cell response to the mitogen Concanavalin A (Con-A) and, after sensitization, to the antigens keyhole limpet haemocyanin (KLH) and human serum albumin (DNP-HSA). In the present investigations we studied the role played by infected macrophages in the development of the cellular unresponsiveness present in visceral leishmaniasis. Adherent spleen cells from infected hamsters were unable to present L. donovani antigens to antigen specific T cells, however they were able to present KLH. Conversely, T cells from infected animals did not respond to parasite antigens even when these antigens were presented by normal syngeneic macrophages. Interestingly, lymphocytes from inguinal lymph nodes of infected animals sensitized in their foot pad with parasite antigens proliferated well when stimulated in vitro with L. donovani antigens. These results suggest that the defect in the cellular immune response of the L. donovani infected hamsters is a consequence of a selective inability of their antigen presenting cells to process and present parasite antigens to T cells.

Cytokine activation of human macrophages infected with HIV-1 to inhibit intracellular protozoa.

Peripheral blood mononuclear cells (PBMCs) from HIV-seronegative donors were infected in vitro with HIV-1. Infection was monitored by cytopathology, supernatant p24 antigen, and by immunocytochemical staining. After 14 days in culture, approximately 70-90% of the cells became infected with HIV, as indicated by cell fusion and immunostaining for virus. At this time, recombinant HuIFN-gamma was added to the cultures, followed by infection 24 h later with the intracellular protozoan parasites Toxoplasma gondii, Trypanosoma cruzi, or Leishmania chagasi. Percentages of intracellular parasites were determined at various points thereafter. Using a system capable of detecting both virus and parasite infection, we determined that (a) cells infected with HIV were capable of ingesting and/or being infected by each of these parasitic protozoa, (b) HIV-infected macrophages could be activated to inhibit the replication of all three parasites following treatment with IFN-gamma, and (c) cultures of HIV-infected macrophages could respond to IFN-gamma with increased oxidative burst activity. The degree of parasite infection or inhibition observed in infected cells was not significantly different from that observed in non-HIV-infected cells. From these observations, we concluded that HIV-1 infection does not render macrophages unresponsive to IFN-gamma activation for microbicidal activity.

Evaluation of the mutagenic potential of the antichagasic drug Rochagan in healthy and chagasic rodents.

Benznidazole (bz) is the active component of the antichagasic drug Rochagan. Tests were carried out to detect the induction of chromosomal aberrations and micronuclei in rodent bone marrow cells and peripheral blood cells, respectively. Rats were exposed to acute treatment with Rochagan by gavage at total doses of 150, 300, 1500, 2000 and 3000 mg bz/kg body weight and killed at different times. In the chronic treatments, healthy and chagasic Balb/c mice were treated with Rochagan by gavage at a dose of 100 mg bz/kg/day for 10 and 25 days. No significant increase in frequency of chromosomal aberrations in bone marrow cells or of micronuclei in peripheral blood cells was detected in the animals acutely or chronically exposed to Rochagan in vivo.

Activity of some naphthoquinones on blood stream forms of Trypanosoma cruzi.

Accidental transmission of Chagas disease to man by blood transfusion is a serious problem in Latin America. This paper describes the testing of several naphthoquinones, some of which were active against blood trypomastigotes in vitro at 4 degrees C and might therefore warrant further study for preventing transmission of Chagas disease by blood transfusion.