The natural compound 7-epiclusianone inhibits superoxide dismutase activity in Schistosoma mansoni.

Schistosomiasis - caused by trematodes from the genus Schistosoma - affects more than 200 million people worldwide. Growing resistance to therapy with praziquantel (PZQ) has encouraged the search for novel treatments against this neglected disease. The compound 7-epiclusianone (7-epi) - isolated from 'bacupari' (the fruit of the Gracinia brasiliensis tree) - has promising activity against Schistosoma mansoni in vitro, damaging the parasite's tegument. However, the target and mechanism of action of 7-epi have not been identified. Here, we examined the possibility that 7-epi harms the tegument by inhibiting parasite superoxide dismutase (SOD), which protects the tegument from damage by reactive oxygen species produced by host immune cells. Molecular docking analysis in silico suggested strong interactions between 7-epi and S. mansoni cytosolic superoxide dismutase (SmCtSOD) at allosteric cavities. Schistosoma mansoni couples were cultivated ex vivo with 12.44-198.96 µm 7-epi for 24 h, and then parasite extracts were tested for lipid peroxidation (as a surrogate for oxidative stress), and SOD activity and expression. Lipid peroxidation levels increased after incubation with concentrations ≥99.48 µm 7-epi, and this compound reduced SOD activity at concentrations ≥24.87 µm. However, contact with 7-epi did not alter SOD expression, by quantitative real-time polymerase chain reaction (qRT-PCR). Our results show that the inhibition of SmCtSOD is partly responsible for the tegument detachment observed after incubation with 7-epi, but is not the only cause of the antiparasitic action of this compound in vitro.

Plasma butyrylcholinesterase activity: a possible biomarker for differential diagnosis between Alzheimer's disease and dementia with Lewy bodies?

Butyrylcholinesterase (BChE) is an enzyme encoded by BCHE gene, responsible for secondary hydrolysis of the acetylcholine. K and -116A BCHE variants were associated with decrease in plasma BChE activity, and their influence has been investigated in diseases with a cholinergic deficit such as Alzheimer's disease (AD) and dementia with Lewy bodies (DLB). In order to check the influence of BCHE genetic variants on enzymatic activity, all patients and controls were genotyped for K and -116A variants. We found lower plasma BChE activity in DLB patients compared to elderly controls and to AD independent of the presence of K or -116A variants. Our results suggest that the reduction of total plasma BChE activity is probably associated with a feedback mechanism and provides a future perspective of using this enzyme as a possible plasmatic marker for differential diagnosis between AD and DLB.

Anxiety-like responses induced by nitric oxide within the BNST in mice: Role of CRF1 and NMDA receptors.

It has been shown that the bed nucleus of the stria terminalis (BNST) of rats contains nitrergic neurons, which are activated during animal exposure to aversive stimuli. The BNST is also populated by glutamatergic and corticotrophin releasing factor (CRFergic) neurons, which in turn are activated under stressful situations. Here we investigated the anxiogenic-like effects of intra-BNST injections of a nitric oxide (NO) donor, NOC-9 in mice. The role of CRFergic and glutamatergic systems on defensive behavior induced by NOC-9 was investigated with previous intra-BNST infusion of different doses of CP376395, a CRF type 1 receptor antagonist (CRF1), or AP-7, an NMDA (N-methyl-D-aspartate) receptor antagonist. Anxiety-like behavior was assessed immediately and 5 min after intra-BNST drug injection, exposing mice to a novel arena and to the elevated plus-maze (EPM; an anxiogenic situation). Results showed that NOC-9 provoked a short period (≈ 150 s) of freezing behavior in the novel arena and increased anxiety in the EPM. Both CP and AP-7 attenuated the anxiogenic-like effects of NOC-9 in the EPM without changing freezing behavior in the novel arena. When given alone (i.e. without prior intra-BNST injection of NOC-9), AP-7 (0.20 nmol), but not CP (0.75, 1.50, or 3.00 nmol), attenuated anxiety in mice exposed to the EPM. These results suggest that CRF1 and NMDA receptors located within the BNST differentially modulate aversive effects induced by NO production in this limbic forebrain structure.

Side of pupillary mydriasis predicts the cognitive prognosis in patients with severe traumatic brain injury.

BACKGROUND: Pupils' abnormalities are associated to bad prognosis in traumatic brain injury. We investigated the association between the side of pupil mydriasis and the long-term cognitive performance of patients with severe traumatic brain injury (TBI). METHODS: We analyzed the cognitive performance of patients admitted at the intensive care unit with isochoric pupils (IP, n = 28), left mydriasis (LM, n = 10), right mydriasis (RM, n = 9) evaluated in mean 2.5 years after the severe TBI and controls (n = 26) matched for age, sex and education level. RESULTS: Patients and controls had similar scores in the four WAIS-III investigated subtests. In comparison with controls, LM patients had lower scores in Letters and Category Fluency and IP patients in Category Fluency. Among the 10 evaluated memory tests, LM patients had lower scores than controls in eight, RM patients in two and IP in three memory tests. IP and RM were 3.5 to nine times more associated to significant impairment (cognitive scores under the percentile 10 of controls) in six of 16 investigated cognitive tests. LM was six to 15 times more associated to significant impairment in 10 of 16 cognitive tests. The association among the pupil abnormalities and cognitive performances remained significant after the multiple linear regression analysis controlling for age, gender, admission coma Glasgow scale and serum glucose, presence of associated trauma, and cranial computed tomography abnormalities. CONCLUSION: Side of admission pupil abnormalities may be a useful variable to improve prognostic models for long-term cognitive performance in severe TBI patients.

The prognostic value of histopathological grading systems in oral squamous cell carcinomas.

OBJECTIVE: This study evaluated the association of four histopathological grading systems (WHO grading system, malignancy grading of the deep invasive margins (MG), histological risk (HR) model, and tumor budding and depth of invasion (BD) model) with clinicopathological parameters and outcome of 113 oral squamous cell carcinomas to identify their roles in prognosis. METHODS: Demographic and clinical features were obtained from patients' records. Sections from all paraffin-embedded blocks were evaluated according to the four grading systems. Demographic and clinical associations were analyzed using chi-square test, and correlations between the grading systems were established with the Spearman's rank correlation test. Survival curves were performed with Kaplan-Meier method, and multivariate analysis based on Cox proportional hazard model was calculated. RESULTS: Significant associations with survival were observed for WHO grading system and BD model in the univariate analysis, but only the BD model was significantly associated with disease outcome as an independent prognostic marker. Age, tumor size, and presence of regional metastasis were also independent markers of reduced survival. CONCLUSION: A significant association between the BD model and outcome of OSCC patients was observed, indicating this new histopathological grading system as a possible prognostic tool.

Atopy patch test with Aleuroglyphus ovatus antigen in patients with atopic dermatitis.

BACKGROUND: Epicutaneous test made with dust mite antigens. OBJECTIVE: Evaluation of the response of the epicutaneous test with Aleuroglyphus ovatus antigen in atopic patients. METHODS: We patch tested 119 individuals, 48 with atopic dermatitis, 50 with respiratory allergy and 21 healthy controls. We compare the positive response frequency to a closed patch test using Aleuroglyphus ovatus antigen in different concentrations and 48 and 96h reading times among those individuals. RESULTS: Six patients with atopic dermatitis (12.5%) and 4 with respiratory atopy (8.0%) had positive reactions. None of the non-atopic controls had a positive response. As the antigen concentration raised, the number of positive reactions to epicutaneous test raised as well. CONCLUSION: Our data suggest a positive relation between Atopy Patch Test positive responses and Aleuroglyphus ovatus antigen concentration, no matter the kind of the atopic clinical expression.

Lip squamous cell carcinoma in a Brazilian population: epidemiological study and clinicopathological associations.

OBJECTIVES: It was evaluated epidemiological aspects of primary lip squamous cell carcinoma (LSCC) and its associations with clinicopathological factors. STUDY DESIGN: This retrospective, cross-sectional study analysed a socio-demographic, clinical, and morphological data of HNSCC in a Brazilian population (n=30). Data analysis included descriptive statistics and bivariate analyses using the chi-square and Fisher 's exact tests to compare the variables. RESULTS: The LSCC represented 10.8% of all oral cavity squamous cell carcinoma. Lip malignant disease was more prevalent in elderly men, with male-to-female ratio of 5:1. Lower lip was more affected. It was observed high rates of chronic solar exposure, and tobacco and alcohol drinking habits. Clinically, early TNM staging, small tumour lesions, and non-metastatic disease were predominant findings. It was identified a high frequency of well differentiated tumor samples. Worse Karnofsky performance status was associated with cervical metastasis. CONCLUSIONS: Our findings showed that LSCC patients exhibited similar epidemiological and clinical profiles as noted in other studies. Still, the occurrence of metastatic disease was associated with a worse physical performance status of the LSCC patients during diagnosis.

Influence of incubation temperature and embryonic motility on the growth of members of Caiman yacare (Daudin, 1802).

This study aimed to evaluate whether skeletal development of the Pantanal Caiman (Caiman yacare) is similarly influenced by temperature variation and controlled increases in embryo motility. All eggs were incubated at 90% humidity and 29 °C for the first 45 days. Thereafter, the incubation temperature was either maintained at 29 °C and embryos were treated with 4-aminopyridine (4-AP) on days 46, 47, 48, and 49 (Group I, 29 °C 4-AP, n = 15); maintained at 29 °C (n = 14; Group II); or at 33 °C (n = 14, Group III). Embryonic movement was measured using an Egg Buddy® digital monitor on days 30, 35, 42, 49, 56, and 60, at which point embryos were euthanized and samples were collected for analysis. No differences were observed between groups with varying incubation temperatures. In contrast, embryonic motility was greater in embryos treated with 4-AP (P < 0.001) on day 49, and this was associated with higher proportions of snout-vent and hand lengths. This study demonstrates for the first time that pharmacologically induced increases in embryo motility result in phenotypic changes to the proportion of elements during prenatal ontogeny, thereby effectively altering the adaptation of the species to specific environments.

Anxiogenesis induced by social defeat in male mice: Role of nitric oxide, NMDA, and CRF1 receptors in the medial prefrontal cortex and BNST.

Nitric oxide (NO) release in the right medial prefrontal cortex (RmPFC) produces anxiogenesis. In the bed nucleus of the stria terminalis (BNST), a region that receives neuronal projections from the mPFC, NO provokes anxiety, an effect that is blocked by local injections of corticotrophin-releasing factor type 1 receptor (CRF1) or n-methyl-d-aspartate receptor (NMDAr) antagonist. Anxiety is also enhanced by social defeat stress, and chronic stress impairs and facilitates, respectively, PFC and BNST roles in modulating behavioral responses to aversive situations. This study investigated whether the (i) chronic social defeat stress (CSDS) increases NO signaling in the mPFC; and/or (ii) anxiogenic effects provoked by the intra-RmPFC injection of NOC-9 (an NO donor) or by CSDS are prevented by intra-BNST injections of AP-7 (0.05 nmol) or CP 376395 (3.0 nmol), respectively, NMDAr and CRF1 antagonists, in male Swiss-Webster mice exposed to the elevated plus-maze (EPM). Results showed that (a) CSDS increased anxiety (i.e., reduced open-arm exploration) and repeatedly activated nNOS-containing neurons, as measured by ΔFosB (a stable nonspecific marker of neural activity) + nNOS double-labeling, in the right (but not left) mPFC, (b) NOC-9 in the RmPFC also increased anxiety, and (c) both CSDS and NOC-9 effects were reversed by injections of AP-7 or CP 376395 into the BNST. These results suggest that NMDA and CRF1 receptors located in BNST play an important role in the modulation of anxiety provoked by NO in the RmPFC, as well as by chronic social defeat in mice.

Flow cytometry analysis of the circulating haemocytes from Biomphalaria glabrata and Biomphalaria tenagophila following Schistosoma mansoni infection.

Aiming to further characterize the haemocyte subsets in Biomphalaria snails, we have performed a detailed flow cytometric analysis of whole haemolymph cellular components using a multiparametric dual colour labelling procedure. Ethidium bromide/acridine orange fluorescence features were used to first select viable haemocytes followed by flow cytometric morphometric analysis based on the laser scatter properties (forward scatter-FSC and side scatter-SSC). Our findings demonstrated that B. glabrata (BG-BH, highly susceptible to S. mansoni) and 2 strains of B. tenagophila (BT-CF, moderately susceptible and BT-Taim, resistant to S. mansoni) have 3 major circulating haemocyte subsets, referred to as small, medium and large haemocytes. The frequency of small haemocytes was higher in BG-BH, while medium haemocytes were the most abundant cell-type in both B. tenagophila strains. Schistosoma mansoni infection resulted in early reduction of large and medium circulating haemocytes followed by an increase of small haemocytes. Although parasite infection induced haemocyte alterations in all Biomphalaria strains, the response was particularly intense in BT-Taim, the parasite-resistant snail. Interestingly, the trematode infection induces changes in haemocytes with less granular rather than in those with more granular profile. The results indicated that, in B. tenagophila of Taim strain, circulating haemocytes, especially the medium and high subset with less granular profile, are very reactive cells upon S. mansoni infection, suggesting that this cell subset would participate in the early parasite destruction observed in this snail strain.

Influence of incubation temperature and embryonic motility on the growth of members of Caiman yacare (Daudin, 1802) / Influência da temperatura de incubação e da motilidade embrionária sobre o crescimento dos membros de Caiman yacare (Daudin, 1802)

Abstract This study aimed to evaluate whether skeletal development of the Pantanal Caiman (Caiman yacare) is similarly influenced by temperature variation and controlled increases in embryo motility. All eggs were incubated at 90% humidity and 29 °C for the first 45 days. Thereafter, the incubation temperature was either maintained at 29 °C and embryos were treated with 4-aminopyridine (4-AP) on days 46, 47, 48, and 49 (Group I, 29 °C 4-AP, n = 15); maintained at 29 °C (n = 14; Group II); or at 33 °C (n = 14, Group III). Embryonic movement was measured using an Egg Buddy® digital monitor on days 30, 35, 42, 49, 56, and 60, at which point embryos were euthanized and samples were collected for analysis. No differences were observed between groups with varying incubation temperatures. In contrast, embryonic motility was greater in embryos treated with 4-AP (P < 0.001) on day 49, and this was associated with higher proportions of snout-vent and hand lengths. This study demonstrates for the first time that pharmacologically induced increases in embryo motility result in phenotypic changes to the proportion of elements during prenatal ontogeny, thereby effectively altering the adaptation of the species to specific environments.

Resumo Este estudo objetivou avaliar os efeitos da temperatura e motilidade embrionária sobre o desenvolvimento esquelético de jacaré-do-pantanal (Caiman yacare). Os ovos foram incubados com 90% de umidade e empregou-se a temperatura de 29°C por 45 dias. Após, para a incubação do Grupo I a temperatura continuou em 29°C, mas associou-se à injeção de 4-aminopiridina (29°C-4AP, n = 15) aplicada nos dias 46, 47, 48 e 49, do Grupo II permaneceu em 29°C (n = 14) e do Grupo III elevou-se para 33°C (n = 14). A movimentação foi mensurada através do monitor digital Egg Buddy® nos dias 30, 35, 42, 49, 56 e 60 dias. Aos 60 dias, os embriões foram eutanasiados e coletadas amostras embrionárias. Na análise estatística não foram observadas diferenças entre os grupos para o fator temperatura sobre a motilidade embrionária no desenvolvimento esquelético. Em contraste, a motilidade evidenciou diferença estatística no dia 49 para o Grupo I (P < 0,001) e apresentou maiores proporções de nariz e mão. Esses dados demonstraram pela primeira vez que o aumento na motilidade, induzidos farmacologicamente resultam em divergências fenotípicas na proporção de segmentos anatômicos durante a ontogenia pré-natal, podendo alterar efetivamente a adaptação dos animais em ambientes específicos.

Influence of incubation temperature and embryonic motility on the growth of members of Caiman yacare (Daudin, 1802)

Abstract This study aimed to evaluate whether skeletal development of the Pantanal Caiman (Caiman yacare) is similarly influenced by temperature variation and controlled increases in embryo motility. All eggs were incubated at 90% humidity and 29 °C for the first 45 days. Thereafter, the incubation temperature was either maintained at 29 °C and embryos were treated with 4-aminopyridine (4-AP) on days 46, 47, 48, and 49 (Group I, 29 °C 4-AP, n = 15); maintained at 29 °C (n = 14; Group II); or at 33 °C (n = 14, Group III). Embryonic movement was measured using an Egg Buddy® digital monitor on days 30, 35, 42, 49, 56, and 60, at which point embryos were euthanized and samples were collected for analysis. No differences were observed between groups with varying incubation temperatures. In contrast, embryonic motility was greater in embryos treated with 4-AP (P 0.001) on day 49, and this was associated with higher proportions of snout-vent and hand lengths. This study demonstrates for the first time that pharmacologically induced increases in embryo motility result in phenotypic changes to the proportion of elements during prenatal ontogeny, thereby effectively altering the adaptation of the species to specific environments.

Resumo Este estudo objetivou avaliar os efeitos da temperatura e motilidade embrionária sobre o desenvolvimento esquelético de jacaré-do-pantanal (Caiman yacare). Os ovos foram incubados com 90% de umidade e empregou-se a temperatura de 29°C por 45 dias. Após, para a incubação do Grupo I a temperatura continuou em 29°C, mas associou-se à injeção de 4-aminopiridina (29°C-4AP, n = 15) aplicada nos dias 46, 47, 48 e 49, do Grupo II permaneceu em 29°C (n = 14) e do Grupo III elevou-se para 33°C (n = 14). A movimentação foi mensurada através do monitor digital Egg Buddy® nos dias 30, 35, 42, 49, 56 e 60 dias. Aos 60 dias, os embriões foram eutanasiados e coletadas amostras embrionárias. Na análise estatística não foram observadas diferenças entre os grupos para o fator temperatura sobre a motilidade embrionária no desenvolvimento esquelético. Em contraste, a motilidade evidenciou diferença estatística no dia 49 para o Grupo I (P 0,001) e apresentou maiores proporções de nariz e mão. Esses dados demonstraram pela primeira vez que o aumento na motilidade, induzidos farmacologicamente resultam em divergências fenotípicas na proporção de segmentos anatômicos durante a ontogenia pré-natal, podendo alterar efetivamente a adaptação dos animais em ambientes específicos.

A remarkable depletion of both naïve CD4+ and CD8+ with high proportion of memory T cells in an IPEX infant with a FOXP3 mutation in the forkhead domain.

IPEX is a rare X-linked syndrome, with immune dysfunction, polyendocrinopathy and enteropathy. We describe an infant who died at the age of 11 months after developing eczema, severe diarrhoea, diabetes, hypothyroidism, thrombocytopenia and four episodes of septicaemia. Immunophenotyping of peripheral blood at 8 months revealed normal CD3+ T, CD4+ T and CD8+ T cell numbers, with low NK and B cells. CD4+ and CD8+ T lymphocytes showed remarkably low numbers and percentages of naïve cells and high numbers of memory CD4 and CD8 cells. At autopsy, an intense depletion of immune cells in thymus, spleen and lymph nodes was observed. No Hassall's corpuscles were found in thymus. Lymphocytic pancreatitis and intense villous atrophy with mucosal lymphocytic infiltration in small bowel were also seen. FOXP3 gene studies revealed a: C-->G substitution 3 bp upstream of exon 10, which prevents splicing between exons 9 and 10, likely resulting in a functionally altered or deficient protein. Florid clinical findings are usually observed in association of forkhead DNA-binding domain mutations. The intense depletion of naïve T cells we report suggest that depletion of immune cells might take place due to uncontrolled activation due to the absence of regulatory T cells.

Participation of cell-free haemolymph of Biomphalaria tenagophila in the defence mechanism against Schistosoma mansoni sporocysts.

Biomphalaria tenagophila of Taim strain is able to completely destroy Schistosoma mansoni sporocyst few hours after parasite penetration, although the mechanism is still not well known. In this experimental work we show that passive transference of cell-free haemolymph, especially from B. tenagophila Taim, resulted in higher resistance of B. tenagophila Cabo Frio to S. mansoni infection. This effect was demonstrated in vivo, by the reduction in the infection rate, and the significantly lower production of sporocysts and cercariae of the parasite in snails treated with Taim cell-free haemolymph compared to CBSS-inoculated snails. The protective effect of Taim cell-free haemolymph was also observed during the in vitro interaction between haemocytes and sporocysts. In this system, addition of B. tenagophila cell-free haemolymph, especially from Taim strain, was responsible for significant increase in sporocyst mortality compared to B. glabrata cell-free haemolymph or culture medium. Moreover, the combination of Taim cell-free haemolymph and Cabo Frio haemocytes increased significantly the mortality of sporocysts. The results show that Taim cell-free haemolymph would act direct and indirectly on destruction of S. mansoni sporocysts. The results also suggest that cell-free haemolymph indirectly increases parasite recognition by the circulating granulocytes and it is species specific.

Effect of Angiostrongylus vasorum infection on Biomphalaria tenagophila susceptibility to Schistosoma mansoni.

Concomitant infection with different parasites may be a helpful laboratorial strategy leading to the better understanding of the mechanisms used by the internal defense system (IDS) of Gastropoda to deal with helminth infection, such as Schistosoma mansoni. This work reports the effect of co-infection of Angiostrongylus vasorum and S. mansoni in hemocyte activity and in the outcome of infection. The simultaneous infection resulted in an increase of snail susceptibility to S. mansoni. In contrast, snails infected with both parasites, 15 days apart, did not show differences in the susceptibility compared to a single parasite infection. The increased susceptibility was measured by the significantly higher number of migrating sporocysts, higher percentage of snails shedding cercariae, higher number of cercariae shed and higher mortality in the experimental group that were simultaneously infected with A. vasorum and S. mansoni, when compared to snails infected only with S. mansoni. Snails simultaneously infected with A. vasorum and S. mansoni showed lower hemocyte activation during the first few days of infection, compared to activation induced only by A. vasorum infection. Between 5 and 15 days post-infection (dpi), granulocyte number and nitric oxide (NO) contents of simultaneously infected snails were lower than the S. mansoni-infected snails. Based on the results, we suggest that differences in the level of hemocyte response could explain the increased S. mansoni susceptibility observed in snails simultaneously infected with both parasites. However, when S. mansoni infection occurred after A. vasorum larvae are completely encapsulated, the response against S. mansoni was not altered, and therefore there were no differences in the susceptibility level.

Physical environment modulates the behavioral responses induced by chemical stimulation of dorsal periaqueductal gray in mice.

In order to investigate the relationship between behaviors elicited by chemical stimulation of the dorsal periaqueductal gray (dorsal PAG) and spontaneous defensive behaviors to a predator, the excitatory amino acid D,L-homocysteic acid (5 nmol in 0.1 micro l), was infused into the dorsal PAG and behavioral responses of mice were evaluated in two different situations, a rectangular novel chamber or the Mouse Defense Test Battery (MDTB) apparatus. During a 1-min period following drug infusion, more jumps were made in the chamber than in the MDTB runway but running time and distance traveled were significantly higher in the runway. Animals were subsequently tested using the standard MDTB procedure (anti-predator avoidance, chase and defensive threat/attack). No drug effects on these measures were significant. In a further test in the MDTB apparatus, the pathway of the mouse during peak locomotion response was blocked 3 times by the predator stimulus (anesthetized rat) to determine if the mouse would avoid contact. Ninety percent of D,L-homocysteic treated animals made direct contact with the stimulus (rat), indicating that D,L-homocysteic-induced running is not guided by relevant (here, threat) stimuli. These results indicate that running as opposed to jumping is the primary response in mice injected with D,L-homocysteic into the dorsal PAG when the environment enables flight. However, the lack of responsivity to the predator during peak locomotion suggests that D,L-homocysteic-stimulation into the dorsal PAG does not induce normal antipredator flight.

No difference in CCND1 gene expression between breast cancer patients with and without lymph node metastasis in a Southern Brazilian sample.

The Cyclin D1 protein has been extensively studied over the last decades, for its various roles in physiological processes, both in normal and cancer cells. Gene amplifications and overexpression of CCND1 are frequently reported in several types of cancers, including breast carcinomas, showing the increasing relevance of Cyclin D1 in tumorigenesis. Little is known about the role of this protein in the metastatic process, and the main objective of this study was to evaluate the importance of the CCND1 as a potential marker of tumor progression in breast carcinomas, in a sample collected in Southern Brazil. We studied 41 samples of formalin-fixed paraffin-embedded tissue sections from invasive ductal breast carcinomas subdivided into metastatic (n = 19) and non-metastatic (n = 22) tumors. Gene expression analysis was performed through Quantitative Real-Time PCR and immunohistochemistry. In spite of the higher expression levels of CCND1 mRNA and protein in tumors when compared with the control samples, no differences were observed between the metastatic and non-metastatic groups, suggesting that, in these samples, the expression of CCND1 has no significant influence on the metastatic process. Further studies must be performed in an attempt to clarify the diagnostic and prognostic value of Cyclin D1 in breast cancers, as well as the mechanisms that trigger its overexpression in tumors.

Functional lateralization of the medial prefrontal cortex in the modulation of anxiety in mice: Left or right?

It has been suggested that the left medial prefrontal cortex (LmPFC) has an inhibitory role in controlling the right mPFC (RmPFC), thereby reducing the deleterious effects of stressors on emotional states. Here, we investigated the effects on anxiety of bilateral or unilateral injections of NOC-9 [a nitric oxide (NO) donor] and cobalt chloride (CoCl2; a synaptic inhibitor) into the mPFC of mice exposed to the elevated plus-maze (Experiments 1 and 2). The effects of restraint or social defeat on anxiety in undrugged mice were recorded at 5 min or 24 h after exposure to the stress (Experiment 3). Experiment 4 investigated the effects of LmPFC injection of CoCl2 combined with restraint or social defeat on anxiety, which was recorded 24 h later. Although intra-RmPFC NOC-9 produced anxiogenesis, its injection into the LmPFC, or bilaterally, did not change anxiety. Intra-RmPFC or -LmPFC injection of CoCl2 produced anxiolytic- and anxiogenic-like effects, respectively. Both restraint and social defeat produced anxiogenesis at 5 min, but defeated mice did not display anxiety 24 h after the stress. Although intra-LmPFC CoCl2 did not change anxiety, which was recorded 24 h later in non-stressed mice, this synaptic inhibitor produced a clear, anxiogenic-like effect in defeated (but not restrained) mice. These results suggest that (i) nitrergic activation of the RmPFC increases anxiety, which in turn is inhibited by NO production within the LmPFC; (ii) neuronal inhibition of the RmPFC or LmPFC elicits anxiolysis and anxiogenesis, respectively; and (iii) inactivation of the LmPFC results in recrudescence of anxiety induced by social defeat stress.

Evaluation of the alcoholic extract of Dipteryx alata Vogel almonds and bark in skin wound healing in C57BL6 mice / [Avaliação do extrato alcoólico da casca e da semente da Dipteryx alata Vogel na cicatrização da pele de camundongos C57BL/6]

The aim of this study was to evaluate the topical application of alcoholic extracts of Dipteryx alata Vogel almonds and bark in skin wound healing in mice. Fifty-four C57BL/6 mice were equally distributed into three groups: Control, Almond, and Bark. A 9 mm skin fragment was resected from the dorsal region of the animals' thorax. The wounds were submitted to topical application of base cream (vehicle), 10% hydroalcoholic almond extract, or bark extract twice a day. Macroscopic, histological, and immunohistochemical evaluations were conducted on the 7th, 14th, and 21st postoperative days. No significant difference was observed regarding skin wound area among groups, with the parameter presenting only a temporal effect on healing (p>0.05). The almond and control groups exhibited more intense collagenization than the bark group (p<0.05). Dipteryx alata Vogel showed to be inert in the wound healing process in mice.(AU)

O objetivo deste estudo foi avaliar a aplicação tópica do extrato alcoólico da semente e da casca da Dipteryx alata Vogel na cicatrização de feridas cutâneas, em camundongos. Um total de 54 camundongos C57BL/6 foram utilizados neste estudo, distribuídos em três grupos de 18 animais (controle, semente e casca). Em todos os animais, um fragmento de pele foi ressecado da região dorsal do tórax utilizando-se instrumento de punção de 9mm de diâmetro, após o qual foi realizada aplicação tópica de creme base (veículo), extrato hidroalcoólico 10% de semente ou casca, duas vezes ao dia. As avaliações macroscópica, histológica e imuno-histoquímica foram realizadas no sétimo, 14º e 21º dias de pós-operatório. Não foi observada diferença significativa quanto à área da ferida cutânea entre os grupos, apenas um efeito temporal na cicatrização (P>0,05), indicando estágio possivelmente mais avançado desse processo. Porém, na avaliação histológica, os grupos semente e controle apresentaram colagenização mais intensa que o grupo casca (P<0,05). Dipteryx alata Vogel mostrou-se inerte no processo de cicatrização de feridas em camundongos.(AU)

Butyrylcholinesterase: K variant, plasma activity, molecular forms and rivastigmine treatment in Alzheimer's disease in a Southern Brazilian population.

Alzheimer's disease (AD) is a neurodegenerative disorder in which there is a decline of cholinergic function. The symptomatic AD treatment involves the use of ChEIs (cholinesterase inhibitors) as rivastigimine, a dual inhibitor. The human butyrylcholinesterase (BChE) is an enzyme that has specific roles in cholinergic neurotransmission and it has been associated with AD. In the serum, BChE is found in four main molecular forms: G1 (monomer); G1-ALB (monomer linked to albumin); G2 (dimer); and G4 (tetramer). The interaction between the products of BCHE gene and CHE2 locus results in CHE2 C5+ and CHE2 C5- phenotypes. CHE2 C5+ phenotype and BChE-K are factors that influence on BChE activity. This work aimed to verify the proportions of BChE molecular forms, total and relative activity in 139 AD patients and 139 elderly controls, taking into account K variant, CHE2 locus, rivastigmine treatment and clinical dementia rating (CDR) of AD patients. Phenotypic frequencies of CHE2 C5+ and frequency of the carriers of the K allele were similar between groups. Total BChE activity in plasma was significantly lower in AD patients than in elderly controls. Furthermore, we found that reduction on plasma BChE activity is associated directly with AD progression in AD patients and that rivastigmine treatment has a stronger effect on BChE activity within the CDR2 group. The reduction in BChE activity did not occur proportionally in all molecular forms. Multiple regression analysis results confirmed that AD acts as the main factor in plasma BChE activity reduction and that severe stages are related with an even greater reduction. These findings suggest that the reduction of total plasma BChE and relative BChE molecular forms activity in AD patients is probably associated with a feedback mechanism and provides a future perspective of using this enzyme as a possible plasmatic secondary marker for AD.