Induction of aggression and anxiety-like responses by perfluorooctanoic acid is accompanied by modulation of cholinergic- and purinergic signaling-related parameters in adult zebrafish.

Perfluorooctanoic acid (PFOA) is a contaminant of global concern owing to its prevalent occurrence in aquatic and terrestrial environments with potential hazardous impact on living organisms. Here, we investigated the influence of realistic environmental concentrations of PFOA (0, 0.25, 0.5, or 1.0 mg/L) on relevant behaviors of adult zebrafish (Danio rerio) (e.g., exploration to novelty, social preference, and aggression) and the possible role of PFOA in modulating cholinergic and purinergic signaling in the brain after exposure for 7 consecutive days. PFOA significantly increased geotaxis as well as reduced vertical exploration (a behavioral endpoint for anxiety), and increased the frequency and duration of aggressive episodes without affecting their social preference. Exposure to PFOA did not affect ADP hydrolysis, whereas ATP and AMP hydrolysis were significantly increased at the highest concentration tested. However, AChE activity was markedly decreased in all PFOA-exposed groups when compared with control. In conclusion, PFOA induces aggression and anxiety-like behavior in adult zebrafish and modulates both cholinergic and purinergic signaling biomarkers. These novel data can provide valuable insights into possible health threats related to human activities, demonstrating the utility of adult zebrafish to elucidate how PFOA affects neurobehavioral responses in aquatic organisms.

Waterborne atenolol disrupts neurobehavioral and neurochemical responses in adult zebrafish.

Environmental contamination by pharmaceuticals from industrial waste and anthropogenic activities poses adverse health effects on non-target organisms. We evaluated the neurobehavioral and biochemical responses accompanying exposure to ecological relevant concentrations of atenolol (0, 0.1, 1.0, and 10 µg/L) for seven uninterrupted days in adult zebrafish (Danio rerio). Atenolol-exposed fish exhibited anxiety-like behavior, characterized by significant bottom-dwelling with marked reduction in vertical exploration. Atenolol-exposed fish exhibited marked increase in the duration and frequency of aggressive events without altering their preference for conspecifics. Biochemical data using brain samples indicated that atenolol disrupted antioxidant enzyme activities and induced oxidative stress. Exposure to atenolol markedly decreased ATP and AMP hydrolysis without affecting ADP hydrolysis and acetylcholinesterase (AChE) activity. Atenolol significantly upregulated tryptophan hydroxylase 1 (tph1) mRNA expression but downregulated brain-derived neurotrophic factor (bdnf) mRNA. Collectively, waterborne atenolol elicits aggressive and anxiety-like responses in adult zebrafish, accompanied by oxidative stress, reduced nucleotide hydrolysis, altered tph1 and bdnf mRNA expression, which may impact the survival and health of fish in aquatic environment.

Bradykinin and its role in osteoarthritis.

Osteoarthritis (OA), the most common joint disorder, is a disease involving all the articular structures. It presents both degenerative and inflammatory aspects. Recently, the important role of Bradykinin (BK), a phlogistic mediator, has been proposed in the pathophysiology of OA. In our review, we summarized the currently available information on the mechanisms of action of BK in OA by linking its B2 receptors. Then, we analyzed the data about the effects of BK in synoviocytes and chondrocytes cultures. Furthermore, we described the action of B2 receptor antagonists (Icatibant and Fasitibant), presenting them as new promising symptom-anddisease- modifying agents in the treatment of OA. However, more in vitro, animal model and clinical studies, are needed to better understand the mechanisms of action as well as the efficacy and tolerability of the B2 receptor antagonists in OA.

Expanding the use of homebase-related parameters to investigate how distinct stressful conditions affect zebrafish behaviors.

Stress is a physiological reaction that allows the organisms to cope with challenging situations daily. Thus, elucidating the behavioral outcomes following different stressors is of great importance in translational research. Here, we aimed to characterize the main factors which explain similarities and differences of two stress protocols on zebrafish exploratory activity. To answer this point, we performed behavioral analyses aiming to simplify the data structure associated with homebase-related measurements in an integrated manner. Adult zebrafish were exposed to conspecific alarm substance for 5 min (acute stress protocol - AS) or submitted to 7 days of unpredictable chronic stress (UCS). Immediately after AS or in the subsequent day following UCS (8th day), fish were individually tested in the open field and the behaviors were recorded for 30 min to posterior identification of homebase locations. For both protocols, behavioral clustering revealed two major clusters, grouping homebase- and locomotor-related parameters, respectively. While AS increased both positive and negative correlations between exploratory and locomotor endpoints, a significant increase in negative correlations was found in UCS-challenged fish. Comparison of the principal component analyses data set revealed a reduced exploratory activity using the homebase in AS group, while decreased locomotion in the periphery and anxiety-like behaviors were evidenced in UCS fish. In conclusion, our findings revealed a different structure of behavior in zebrafish following AS and UCS protocols, supporting the existence of distinct behavioral strategies to cope with acute and chronic stress. Furthermore, we expand the use of homebase-related measurements as a valuable tool to investigate complex behavioral modulations in future translational neuropsychiatry research.

Acetic acid-induced pain elicits stress-, and camouflage-related responses in zebrafish: Modulatory effects of opioidergic drugs on neurobehavioral phenotypes.

While pain results from the activation of nociceptors following noxious stimuli, mounting evidence links pain- and stress-related responses in mammals. In zebrafish, the activation of hypothalamic-pituitary-interrenal (HPI) axis may also regulate body pigmentation (the camouflage response). Here, we aimed to investigate a putative relationship between pain-, stress-, and camouflage-related parameters in adult zebrafish. To answer this question, we assessed whether intraperitoneal acetic acid injection can activate the HPI axis, measuring whole-body cortisol and the camouflage response as physiological endpoints in the presence or absence of morphine or naloxone, an opioid antagonist. Acetic acid induced a stereotypic circling behavior in the top of the tank, accompanied by abdominal writhing-like response, a specific phenotype that reflects local nociceptive effect. Both whole-body cortisol levels and camouflage response increased in the acetic acid group, while morphine prevented these responses, and naloxone antagonized morphine-induced effects. Moreover, we observed positive correlations between representative behavioral, physiological and skin coloration endpoints, and a "pain index" was proposed to summarize phenotypic profile of zebrafish under different pharmacological manipulations. Collectively, these findings suggest a coordinated activation of pain, camouflage- and stress-related pathways following acetic acid injection in zebrafish. Our data also support that camouflage response represents a novel and relevant biomarker for future probing pain and stress neurobiology, with a robust sensitivity to opioidergic drugs.

Acetic acid-induced nociception modulates sociability in adult zebrafish: Influence on shoaling behavior in heterogeneous groups and social preference.

Due to the recognition of fishes as sentient beings, the zebrafish (Danio rerio) has become an emergent animal model system to investigate the biological processes of nocifensive responses. Here, we aimed to characterize the zebrafish social behavior in a nociception-based context. For this purpose, using a three-dimensional analysis of heterogeneous shoals, we investigated the main behavioral responses in two 6-min trials: before (baseline) and after a single intraperitoneal (i.p) injection of 10 µL phosphate-buffered saline (PBS) (control), acetic acid 5% (AA), morphine 2.5 mg/kg (MOR) or acetic acid 5% plus morphine 2.5 mg/kg (AA + MOR) in one subject from a four-fish shoal. The social preference of individuals for tanks with shoals of fish treated with PBS, 5 % AA, or to an empty aquarium was also tested. We verified that AA administration disrupted the shoal homogeneity by eliciting dispersion of the treated fish with simultaneous clustering of non-manipulated fish. In general, morphine coadministration protected against AA-induced behavioral changes. The social preference test revealed a clear preference to conspecifics (PBS and AA) over an empty tank. However, a prominent preference for PBS- over AA-treated shoal was verified. Overall, our novel findings show that nociception can modulate zebrafish sociability, possibly due to the visual recognition of nocifensive responses. Although future studies are needed to elucidate how nociception modulates zebrafish social behavior, our results contribute to improve the welfare assessment of zebrafish shoals under distinct experimental manipulations.

Zebrafish as a potential non-traditional model organism in translational bipolar disorder research: Genetic and behavioral insights.

Bipolar disorder (BD) is a severe and debilitating illness that affects 1-2% of the population worldwide. BD is characterized by recurrent and extreme mood swings, including mania/hypomania and depression. Animal experimental models have been used to elucidate the mechanisms underlying BD and different strategies have been proposed to assess BD-like symptoms. The zebrafish (Danio rerio) has been considered a suitable vertebrate system for modeling BD-like responses, due to the genetic tractability, molecular/physiological conservation, and well-characterized behavioral responses. In this review, we discuss how zebrafish-based models can be successfully used to understand molecular, biochemical, and behavioral alterations paralleling those found in BD. We also outline some advantages and limitations of this aquatic species to examine BD-like phenotypes in translational neurobehavioral research. Overall, we reinforce the use of zebrafish as a promising tool to investigate the neural basis associated with BD-like behaviors, which may foster the discovery of novel pharmacological therapies.

Utility of cockroach as a model organism in the assessment of toxicological impacts of environmental pollutants.

Environmental pollution is a global concern because of its associated risks to human health and ecosystem. The bio-monitoring of environmental health has attracted much attention in recent years and efforts to minimize environmental contamination as well as to delineate toxicological mechanisms related to toxic exposure are essential to improve the health conditions of both humans and animals. This review aims to substantiate the need and advantages in utilizing cockroaches as a complementary, non-mammalian model to further understand the noxious impact of environmental contaminants on humans and animals. We discuss recent advances in neurotoxicology, immunotoxicology, reproductive and developmental toxicology, environmental forensic entomotoxicology, and environmental toxicology that corroborate the utility of the cockroach (Periplaneta americana, Blaptica dubia, Blattella germanica and Nauphoeta cinerea) in addressing toxicological mechanisms as well as a sensor of environmental pollution. Indeed, recent improvements in behavioural assessment and the detection of potential biomarkers allow for the recognition of phenotypic alterations in cockroaches following exposure to toxic chemicals namely saxitoxin, methylmercury, polychlorinated biphenyls, electromagnetic fields, pharmaceuticals, polycyclic aromatic hydrocarbon, chemical warfare agents and nanoparticles. The review provides a state-of-the-art update on the current utility of cockroach models in various aspects of toxicology as well as discusses the potential limitations and future perspectives.

Earthworm assemblages in different intensity of agricultural uses and their relation to edaphic variables.

The objective of this study was to relate earthworm assemblage structure with three different soil use intensities, and to indentify the physical, chemical, and microbiological soil variables that are associated to the observed differences. Three soil uses were evaluated: 1-Fifty year old naturalized grasslands, low use intensity; 2-Recent agricultural fields, intermediate use intensity, and 3-Fifty year old intensive agricultural fields, high use intensity. Three different sites for each soil use were evaluated from winter 2008 through summer 2011. Nine earthworm species were identified across all sampling sites. The sites shared five species: the native Microscolex dubius, and the introduced Aporrectodea caliginosa, A. rosea, Octalasion cyaneum, and O. lacteum, but they differed in relative abundance by soil use. The results show that the earthworm community structure is linked to and modulated by soil properties. Both species abundance and diversity showed significant differences depending on soil use intensity. A principal component analysis showed that species composition is closely related to the environmental variability. The ratio of native to exotic species was significantly lower in the intensive agricultural system when compared to the other two, lower disturbance systems. Microscolex dubius abundance was related to naturalized grasslands along with soil Ca, pH, mechanical resistance, and microbial respiration. Aporrectodea caliginosa abundance was related to high K levels, low enzymatic activity, slightly low pH, low Ca, and appeared related to the highly disturbed environment. Eukerria stagnalis and Aporrectodea rosea, commonly found in the recent agricultural system, were related to high soil moisture condition, low pH, low Ca and low enzymatic activity. These results show that earthworm assemblages can be good indicators of soil use intensities. In particular, Microscolex dubius, Aporrectodea caliginosa, and Aporrectodea rosea, showed different temporal patterns and species associations, due to the changes in soil properties attributable to soil use intensity, defined as the amount and type of agricultural operations.

A dielectric inverse problem applied to human skin measurements during glucose excursions.

A fringing field capacitive sensor has been used to measure the dielectric properties of human skin and underlying tissue in the MHz frequency range. It has recently been shown in clinical experimental studies that these dielectric properties can be related to the effects of in vivo glucose variations of the test subject. Previously, the relationship between electrical impedance and the glucose level has been established via statistical methods, such as the regression method. In this work, we explored a different approach, namely the resolution of the so-called inverse problem. First we applied the method on an artificial two-layer lossy system in order to test the sensitivity of the solution to forced changes in the layer properties and its stability to a constant setting. After validation of this method on artificial systems, a similar inverse problem was set and solved for dielectric measurements on human skin during an induced glucose excursion, where the skin is also modelled as a double-layer system. The changes of the measured permittivity and conductivity of the second layer versus the glucose changes are calculated for 22 study days. The statistical distribution shows that the median slopes of both dielectric properties are negative. These results can be used to test our hypothesis and to continue building potential explanations for the phenomena induced by the glucose changes on the skin layer dielectric parameters.

Echography of optic disc related pathology in diabetic retinopathy.

Combined A- and B-scan examination of eyes with diabetic retinopathy and opaque media provides information about the presence and density of vitreous hemorrhages, can detect membranes inserting into the optic disc or the retina and differentiate them from retinal detachment. Absolute and relative indications for echographic examination are listed and some echograms concerning vitreous hemorrhage, proliferating membranes and traction retinal detachment are presented. The value of both A-scan standardized echography and contact B-scan technique for previtrectomy evaluation is stressed.

Disposable film cover for the tip of Goldmann's tonometer.

The application of a thin, transparent, polyvinylchloride film on the double prism of Goldmann's tonometer is proposed, so as to prevent bacterial and viral infection during tonometry. This film, mounted in such a way as to facilitate its application, is supplied in disposable sterile packaging. The use of the film does not alter the calibration of the instrument and does not substantially modify readings thereby obtained.

Self-administered pain-relieving manoeuvres in primary headaches.

We investigated the use of self-administered pain-relieving manoeuvres on a sample of 400 patients with primary headaches--represented by an even distribution of migraine without aura (MO), migraine with aura (MA), episodic tension-type headache (TH), and cluster headache (CH)--consecutively seen at Padua and Parma Headache Centres. Manoeuvres on various regions of the head were used by 258 patients (65% of the cases). The most applied procedures were: compression (114 out of 382 manoeuvres; 30%), application of cold (27%), massage (25%) and application of heat (8%). A significant (P < 0.001) relationship was found between headache diagnoses and type of manoeuvre. In MO patients the application of cold (38% of the manoeuvres) and compression (36%), used mainly on the forehead and temples, prevailed; compression, mainly on the temples, was the most frequent procedure (44%) in MA patients. Massage on the temples and nape was the predominant manoeuvre (43%) in TH patients, whereas in the CH group, which more often required heterogeneous procedures, none of the above-mentioned manoeuvres was prevalent. Compression, as a diagnostic criterion for MO, had a sensitivity of 33% and a specificity of 86%; for the application of cold the figures were 36% and 84%, respectively. Massage had a sensitivity of 33% and a specificity of 80% for TH. The efficacy of the self-administered manoeuvres in reducing pain was scarce. Only 8% of the manoeuvres, in fact, resulted in a good or excellent pain control. Moreover, the efficacy of the manoeuvre was often momentary, wearing off when the manoeuvre stopped. In spite of this, 46% of the subjects used the manoeuvres constantly, at each attack.

Complete migration of retained surgical sponge into ileum without sign of open intestinal wall.

A 24-year-old woman came to the emergency room with a history of diffuse abdominal pain in the form of colic, nausea, vomiting and intestinal constipation. Clinical and ultrasound findings suggested intestinal obstruction due to foreign body. She had been submitted to a cesarean section 4 months previously at another hospital. At laparotomy, a ileum loop was found to be distended by an inside large and hardened mass with another intestinal loops and omentum density adherent. An ileotomy was performed on the compromised segment with terminating anastomosis. When opened surgical specimen it was observed an intraluminal surgical sponge that had completely migrated into the interior of the ileum and stopped next to ileumcecal valve. No fistulas or open intestinal wall were observed.

Our experience in the diagnosis of intraocular tumours by a B-scan computerized system and angiodynography (Doppler). Preliminary results.

The authors compare the ultrasound diagnostic results of intraocular tumours by A/B-scans (Sonomed B 3000, Sonocare, Sonovision STT-100) with images obtained using computerized B-scan (Sonocare, Sonovision STT-100, Acoustic Tissue Typing ATT) and angiodynopgraphy systems (Quantum Philips). The Sonovision uses a computerized ultrasound spectrum analysis to assess the probability that a given lesion is a certain tumour, rather than another. The ATT system provides diagnostic probability for type B and type E melanoma, for haemangioma and metastatic carcinoma. The Quantum ultrasound equipment was developed for studying the heart and the major blood vessels. It is a colour Doppler that simplifies the Doppler technique, allowing it to study small anatomical parts such as tumour-like lesions of the eye. The Doppler technique ascertains the presence and the velocity of blood flow in the tumours. In presenting the preliminary results with the new techniques the authors are aware that the ATT system is not designed for some of the lesions under study (melanomas after conservative radiotherapy).