Effects of short-term exposure to genistein and overfeeding diet on the neural and retinal progenitor competence of adult zebrafish (Danio rerio).

Neurogenesis is a process that occurs throughout the life of a vertebrate. Among the different factors that may affect the natural occurrence of neurogenesis, obesity seems to decrease the proliferation capacity of progenitor neuronal cells. Conversely, the phytoestrogen genistein is known to attenuate some obesity effects beyond its neuroprotective action. Aiming to improve the understanding of how obesity and genistein trigger an impact on the neural and retinal progenitor competence of adult zebrafish, fish were exposed to genistein (GEN - 2 µg L-1) alone or combined with two dietary groups (control and overfeed - OFD) for up to 9 weeks. Zebrafish were fed once per day with Artemia sp. in the control and GEN (2% of BW, control diet), and three times per day in the OFD and OFD + GEN groups (12% BW, overfeeding diet). To assess obesity induction, BMI, biometric parameters, and PPAR-γ protein were quantified. Afterwards, qRT-PCR and immunohistochemistry were performed to determine the cell proliferation and the presence of stem cells through PCNA and Sox-2. Our findings proved that overfeeding adult zebrafish increased the general growth and induced the development of fatty liver. However, for OFD + GEN, this effect was assuaged through the anti-adipogenic effect of GEN. This finding suggests that phytoestrogens could be beneficial to reduce the negative effects of obesity. Moreover, OF induced negative effects on retinal and brain homeostasis, decreasing the proliferation capacity of progenitor neuronal cells. With regard to retinal progenitor competence, genistein seems to mitigate the negative impacts of obesity, whereas the effects of obesity on the brain were exacerbated by this phytoestrogen which negatively influenced the homeostasis of zebrafish neural progenitor competence. This study highlighted the fact that the effects of phytoestrogens in adult neural progenitor competence are complex and could exhibit dissimilar effects depending on the tissue.

Molecular Profile of Sensitization to Dermatophagoides pteronyssinus Dust Mite in Portugal.

BACKGROUND AND OBJECTIVES: To analyze component-resolved diagnosis of sensitization to Dermatophagoides pteronyssinus (Der p) in patients with respiratory allergy and the association between diagnostic findings and clinical severity in different geographical areas. METHODS: The study population comprised 217 patients (mean age, 25.85 [12.7] years; 51.16% female) selected from 13 centers in Portugal (5 from the North, n=65). All had allergic rhinitis with or without asthma and positive skin prick test results to at least 1 dust mite. Specific IgE (sIgE) to Der p, Dermatophagoides farinae, Lepidoglyphus destructor, Der p 1, Der p 2, Der p 10, and Der p 23 was determined using ImmunoCAP. The Mann-Whitney test was applied for the following comparisons: rhinitis vs rhinitis and asthma; mild vs moderate-to-severe rhinitis; North vs South. RESULTS: The prevalence of sensitization was 98.2% for Der p, and 72.4%, 89.4%, 9.7%, and 77% for Der p 1, Der p 2, Der p 10, and Der p 23, respectively. The corresponding median sIgE levels were 8.56, 17.7, 0.01, and 3.95 kUA/L. sIgE to all allergens was higher in patients with moderate-to-severe rhinitis and rhinitis with asthma (nonsignficant). Concentrations of sIgE to Der p 2 were significantly higher in the South than in the North (P=.0496). CONCLUSION: The most common sensitization in Portugal was to Der p. The highest prevalence and median sIgE level were observed for Der p 2. All sIgE values for molecular components were higher in more symptomatic patients (nonsignificant). Concentrations of sIgE to Der p 2 were higher in the South, probably because of the warmer temperature and/or the larger sample size.

Epidemiologic analysis of salivary gland tumors over a 10-years period diagnosed in a northeast Brazilian population.

BACKGROUND: Salivary gland tumors (SGT) correspond to a heterogeneous group of lesions with variable biological behavior. The present study aimed to determine the distribution and demographic findings of salivary gland neoplasms in a northeast Brazilian population. MATERIAL AND METHODS: A retrospective descriptive cross-sectional study was performed. A total of 588 cases of SGT were diagnosed between 2006 and 2016 of 4 pathology services in the state of Sergipe, Brazil. All cases were reviewed, and data such as sex, age, anatomical location, and histopathological diagnosis were collected. RESULTS: A total of 470 (79.9%) tumors were benign and 118 (20.1%) were malignant. The majority of the patients were females (n=328, 55.8%) with an overall female:male ratio of 1.2:1. The major salivary glands were affected more than the minor glands (69.5% vs. 30.5%). Pleomorphic adenoma (n=419, 71.3%) and mucoepidermoid carcinoma (n=29, 4.9%) were the most frequent benign and malignant tumors, respectively. In addition, both benign and malignant tumors occurred more frequently in the parotid gland (n=300, 51%, p<0.05). CONCLUSIONS: The epidemiologic profile and clinical characteristics of SGT were similar to those described in other countries and other regions of Brazil. Epidemiological studies of SGT help to understand their clinical and pathological features and are essential to establish the proper management and prognosis.

MoS2 flakes stabilized with DNA/RNA nucleotides: In vitro cell response.

Two-dimensional transition metal dichalcogenides (TMDCs), such as MoS2 and WS2, have recently emerged as nanomaterials with potential use in biomedicine. An attractive means to favor their interaction with biological media is the use of proper biomolecules as exfoliating/dispersing agents. Here, MoS2 flakes were stabilized with different small functional biomolecules such as adenosine monophosphate (AMP), guanosine monophosphate (GMP) and flavin mononucleotide (FMN) through the strong nucleotide-MoS2 interaction of Lewis acid-base type, rather than just on the weak dispersive and hydrophobic forces commonly associated with the use of many surfactants. The impact of the nucleotide-stabilized MoS2 flakes on the viability and cell proliferation, on the production of intracellular reactive oxygen species (ROS), and on the preosteoblast differentiation process (early stage) has been also evaluated, as well as the incorporation and intracellular localization of the nanomaterials by MC3T3-E1 and Saos-2 cells. The nucleotide-stabilized MoS2 flakes were found to exhibit excellent biocompatibility. Furthermore, their incorporation did not affect the integrity of the cell plasma membrane, which makes them ideal candidates for delivering drug/gene directly into cells. The in vitro cell response of tumor cells to these nanomaterials differs from that of undifferentiated cells, which provides the basis for their potential use in cancer therapy.

Multicentric cutaneous keratoacanthomas in a free-living marmoset (Callithrix sp.).

Cutaneous neoplasia is common in non-human primates. We describe the gross and microscopic features of multicentric cutaneous keratoacanthomas in a free-living marmoset (Callithrix sp.). Immunohistochemistry for human papillomavirus and herpes simplex virus type I and simplex virus type II was negative. Keratoacanthomas should be included in the differential diagnosis for cutaneous masses in non-human primates.

Parotid Salivary Gland Basal Cell Adenocarcinoma in a Big-eared Opossum (Didelphis aurita).

The opossum (family Didelphidae) is a marsupial endemic to the Americas. Apart from the South American short-tailed opossum (Monodelphis domestica) and the Virginia opossum (Didelphis virginiana), there is considerable lack of knowledge about the health and diseases of most opossum species. Among these, the big-eared opossum (Didelphis aurita) is found in Argentina, Brazil and Paraguay. Natural and experimental studies have shown this species to be susceptible to infectious agents with zoonotic potential and the animals may play a role in transmission of such agents. However, neoplasia appears to be uncommon in this species. We describe the gross, microscopical and immunohistochemical features of a parotid salivary gland basal cell adenocarcinoma in a free-living big-eared opossum. This case represents the first report of salivary gland neoplasia in opossums.

Hepatocellular carcinoma in a free-living marmoset (Callithrix sp.) with concomitant biliary trematodiasis.

Hepatocellular carcinoma (HCC) is rare in New World primates. We report the gross, microscopical, and immunocytochemical features of a spontaneous HCC in a free-living marmoset (Callithrix sp.). Hepatitis B and C virus and aflatoxin immunohistochemistry were negative; however, concomitant intra- and extrahepatic biliary trematodiasis could have played a role.

The Vespid Allergy Quality of Life Questionnaire - cultural adaptation and translation to Portuguese.

SUMMARY: A cross-cultural translation of the Vespid Allergy Quality of Life Questionnaire (VQLQ) to the Portuguese population (VQLQ-P) was performed, assessing its applicability in wasp and in non-beekeeper bee venom allergic patients. Additionally, we evaluated a Visual Analogue Scale (VAS) to estimate hymenoptera allergy interference with daily life. Methods. Cross-cultural translation was performed according to recommendations. The final VQLQ-P version, the Expectation of Outcome questionnaire (EoQ), EQ-5D and VAS were applied to wasp (n = 19) and non-beekeeper bee venom allergic patients (n = 30). Results. VQLQ-P significantly correlated with EoQ, (r = 0.76, p < 0.01), EQ-5D (usual activities and anxiety / depression dimensions) and VAS, with a good internal consistency (Cronbach α = 0.88) in wasp allergic individuals. VQLQ-P and EoQ correlation was also high (r = 0.67, p < 0.01) in bee allergy. Conclusion. The VQLQ-P is a valuable tool to evaluate quality of life impairment in Portuguese hymenoptera venom allergic individuals.

Magnetic resonance enterography is feasible and reliable in multicenter clinical trials in patients with Crohn's disease, and may help select subjects with active inflammation.

BACKGROUND: Reliable tools for patient selection are critical for clinical drug trials. AIM: To evaluate a consensus-based, standardised magnetic resonance enterography (MRE) protocol for selecting patients for inclusion in Crohn's disease (CD) multicenter clinical trials. METHODS: This study recruited 20 patients [Crohn's Disease Activity Index (CDAI) scores: <150 (n = 8); 150-220 (n = 4); 220-450 (n = 8)], to undergo ileocolonoscopy and two MREs (with and without colonic contrast) within a 14-day period. Procedures were scored centrally using, Magnetic Resonance Index of Activity (MaRIA), and both Crohn's Disease Endoscopic Index of Severity (CDEIS) and Simplified Endoscopic Score (SES-CD). RESULTS: 37 MREs were acquired. Both MREs were evaluable in 16 patients for calculation of test-retest and inter-reader reliability scores. The MaRIA scores for the terminal ileum had excellent test-retest and inter-reader reliability, with correlations >0.9. The proximal ileum showed strong within-reader agreement (0.90-0.96), and fair between-reader agreement (0.59-0.72). MRE procedures were tolerable. MaRIA scores correlated with CDEIS and SES-CD (0.63 and 0.71), but not with CDAI (0.34). MRE identified 3 patients with intra-abdominal complications, who would otherwise have been included in clinical trials. Furthermore, both MRE and ileocolonoscopy identified active bowel wall inflammation in 2 patients with CDAI <150, and none in 1 patient with CDAI > 220. Data quality was good/excellent in 85% of scans, and fair or better in 96%. CONCLUSIONS: Magnetic resonance enterography of high-quality and reproducibility was feasible in a global multi- centre setting, with evidence for improved selectivity over CDAI and ileocolonoscopy in identifying appropriate CD patients for inclusion in therapeutic intervention trials.

Assessment of asthma control: clinical, functional and inflammatory aspects.

BACKGROUND: Asthma is a complex disease with numerous markers of severity/activity. Clinical assessment,functional parameters and inflammation biomarkers are the most used A correlation between them is difficult, as each one evaluates a particular aspect of the disease. OBJECTIVE AND METHODS: To explore the possible association between asthma control, pulmonary function and inflammation in patients with asthma, consecutive asthmatics underwent simultaneous spirometry (measurement of FEV1), exhaled nitric oxide (eNO) evaluation and Asthma Control Test (ACTTM) questionnaire. RESULTS: The study included 232 asthmatics (mean age: 37.48 years; 78.4%female): 43% had uncontrolled asthma (ACTTM < or = 19) with FEV1 mean values of 83.3% +/- 21.8; 48% partially controlled (ACTTM: 20-24) with FEV1 of 87.6% +/- 17; 9% complete control (ACTTM = 25) with FEV1 of 93.1 +/- 20.6. The relation ACTTM/FEV1 and ACTTM/FEF 25-75% was statistically significant (p = 0.001 and p = 0.034, respectively). Among patients with eNO < 35 ppb, 66% had FEV1 > 80% and 52% had ACTTM > 19. No association was found combining ACTTM/eNO or FEV1/eNO. A subgroup of 66 patients was evaluated twice. CONCLUSION: An association was found between ACTTM and spirometry, with higher ACTTM scores reflecting less bronchial obstruction. The authors advise a combined approach in asthma follow-up, involving clinical aspects, functional parameters and inflammation biomarkers, although in some circumstances ACT could be a valid instrument to be used alone to assess control.

Gill histopathological and oxidative stress evaluation in native fish captured in Portuguese northwestern rivers.

The Northwestern Portuguese region is densely populated and highly industrialized, suffering from high anthropogenic pressure. To assess the biological effect of the several pollutants that are constantly released to the water, a biomarker-based biomonitoring is a promising approach that may provide early-warning signals of pollutants exposure. Fish gill is the first target of pollutants action, thus histopathological and biochemical changes may constitute potential biomarkers. To evaluate this hypothesis, three native fish species (barbel-Luciobarbus bocagei, chub-Squalius carolitertii and nase-Pseudochondrostoma sp.) were sampled in Northwestern Portuguese rivers, the gill histopathological changes were qualitative and quantitatively analyzed and the lipid peroxidation and glutathione-S-transferase activity were determined. A multivariate statistical analysis was performed to establish correlations between these biological responses, environmental variables and ecological status. The quantitative evaluation of the main histopathological changes and oxidative stress responses emphasize the differences, among species, in the responses to the presence of contaminants in water. Discriminant canonical analysis showed that filament epithelium proliferation, necrosis and GST activity were the main contributors to discriminate the ecological status classification. In addition, the results showed that a wide range of environmental factors are influencing fish physiology. In conclusion, the gill biological responses, although not reflecting specific contaminants, can be used as biomarkers of ecosystems perturbation.

Effect of the hydrostatic pressure on otolith growth of early juveniles of Nile tilapia Oreochromis niloticus.

Nile tilapia Oreochromis niloticus early juveniles were maintained for 2 weeks in a pressurized system under a controlled photoperiod, at constant salinity and temperature. Groups of fish were exposed to one of three absolute hydrostatic pressure (HP) regimes: (1) a constant normal atmospheric pressure (100 kPa), (2) a constant 40 m pressure (500 kPa) or (3) a semi-diurnal cyclic vertical migration (100-500 kPa). No significant differences were detected in otolith size and incremental periodicity among the three HP treatments, suggesting that HP does not affect otolith growth of early juveniles O. niloticus.

A new osteophyte segmentation algorithm using partial shape model and its applications to rabbit femur anterior cruciate ligament transection via micro-CT imaging.

Osteophyte is an additional bony growth on a normal bone surface limiting or stopping motion at a deteriorating joint. Detection and quantification of osteophytes from CT images is helpful in assessing disease status as well as treatment and surgery planning. However, it is difficult to distinguish between osteophytes and healthy bones using simple thresholding or edge/texture features due to the similarity of their material composition. In this paper, we present a new method primarily based active shape model (ASM) to solve this problem and evaluate its application to anterior cruciate ligament transection (ACLT) rabbit femur model via CT imaging. The common idea behind most ASM based segmentation methods is to first build a parametric shape model from a training dataset and apply the model to find a shape instance in a target image. A common challenge with such approaches is that a diseased bone shape is significantly altered at regions with osteophyte deposition misguiding an ASM method and eventually leading to suboptimum segmentations. This difficulty is overcome using a new partial ASM method that uses bone shape over healthy regions and extrapolates it over the diseased region according to the underlying shape model. Finally, osteophytes are segmented by subtracting partial-ASM derived shape from the overall diseased shape. Also, a new semi-automatic method is presented in this paper for efficiently building a 3D shape model for an anatomic region using manual reference of a few anatomically defined fiducial landmarks that are highly reproducible on individuals. Accuracy of the method has been examined on simulated phantoms while reproducibility and sensitivity have been evaluated on CT images of 2-, 4- and 8-week post-ACLT and sham-treated rabbit femurs. Experimental results have shown that the method is highly accurate ( R2 = 0.99), reproducible (ICC = 0.97), and sensitive in detecting disease progression (p-values: 0.065,0.001 and < 0.001 for 2- vs. 4, 4- vs. 8- and 2- vs. 8-weeks, respectively).

SVR versus neural-fuzzy network controllers for the sagittal balance of a biped robot.

The real-time balance control of an eight-link biped robot using a zero moment point (ZMP) dynamic model is difficult due to the processing time of the corresponding equations. To overcome this limitation, two alternative intelligent computing control techniques were compared: one based on support vector regression (SVR) and another based on a first-order Takagi-Sugeno-Kang (TSK)-type neural-fuzzy (NF) network. Both methods use the ZMP error and its variation as inputs and the output is the correction of the robot's torso necessary for its sagittal balance. The SVR and the NF were trained based on simulation data and their performance was verified with a real biped robot. Two performance indexes are proposed to evaluate and compare the online performance of the two control methods. The ZMP is calculated by reading four force sensors placed under each robot's foot. The gait implemented in this biped is similar to a human gait that was acquired and adapted to the robot's size. Some experiments are presented and the results show that the implemented gait combined either with the SVR controller or with the TSK NF network controller can be used to control this biped robot. The SVR and the NF controllers exhibit similar stability, but the SVR controller runs about 50 times faster.

Disruption of zebrafish (Danio rerio) embryonic development after full life-cycle parental exposure to low levels of ethinylestradiol.

Exposure of fish to the synthetic estrogen ethinylestradiol (EE2) has been shown to induce a large set of deleterious effects. In addition to the negative impact of EE2 in reproductive endpoints, concern has recently increased on the potential effects of EE2 in fish embryonic development. Therefore, the present study aimed at examining the effects of EE2 on the full embryonic development of zebrafish in order to identify the actual phases where EE2 disrupts this process. Hence, zebrafish were exposed to environmentally relevant low levels of EE2, 0.5, 1 and 2ng/L (actual concentrations of 0.19, 0.24 and 1ng/L, respectively) from egg up to eight months of age (F(1)), and the survival as well as the occurrence of abnormalities in their offsprings (F(2)), per stage of embryonic development, was investigated. A thorough evaluation of reproductive endpoints and transcription of vtg1 gene in the parental generation (F(1)) at adulthood, was performed. No significant differences could be observed for the two lowest EE2 treatments, in comparison with controls, whereas vtg1 transcripts were significantly elevated (40-fold) in the 2ng/L EE2 treatment. In contrast to the findings in the F(1) generation,a significant concentration-dependent increase in egg mortality between 8 and 24hours post-fertilization (hpf) was observed for all EE2 treatments, when compared with controls. The screening of egg and embryo development showed a significant increase in the percentage of abnormalities at 8 hpf for the highest EE2 concentration, a fact that might explain the increased embryo mortality at the 24 hpf time-point observation. Taken together, these findings indicate that the two lowest tested EE2 concentations impact late gastrulation and/or early organogenesis, whereas exposure to 2ng/L EE2 also disrupts development in the blastula phase. After early organogenesis has been completed (24 hpf), no further mortality was observed. These results show that increased embryo mortality occurs at EE2 levels below those inducing reproductive impairment and vtg1 gene induction in the male parental generation, thus suggesting that EE2 may impact some fish populations at levels below those inducing an increase in vtg1 transcripts. Hence, these findings have important implications for environmental risk assessment, strongly supporting the inclusion of embryonic development studies in the screening of endocrine disruption in wild fish populations.