Behavioral expression of emotions and non-invasive assessment of physiological correlates in dogs (Canis familiaris)

The aim of this thesis was to validate the use of infrared thermography (IRT) to non-invasively measure emotional reactions to different situations in pet dogs (Canis familiaris). A preliminary test, aimed to evaluate the correlation between eye-temperature and rectal temperature in dog, was performed. Then, in three different situations, negative (veterinary visit), positive (palatable food rewards), and mildly stressing followed by mildly positive (separation from and reunion with the owner), variations in heat emitted from lacrimal caruncle (referred to as eye temperature) were measured with an infrared thermographic camera. In addition, heart rate and heart rate variability parameters were collected using a non-invasive heart rate monitor designed for human use and validated on dogs. All experiments were video recorded to allow behavioral coding. During the negative situation dogs’ level of activity and stress related behaviors varied across compared to the baseline and dogs showed an increase in eye temperature despite having a significant decrease in the level of activity. The positive situation was characterized by a peak in eye temperature and mean HR and dogs engaged in behaviors indicating a positive arousal, focusing on food treats and tail wagging but there were not variations in HRV during stimulation but only an increment in SDNN immediately after the stimulus. In the separation from and reunion with the owner dogs’ eye temperature and mean HR did not vary neither in the stressful nor in the positive situations, RMSSD increased after the positive episode, SDNN dropped during the two stimulations and it increased after the stimulations. During the separation from the owner dogs were mainly directed to the door or to the experimenter while during the reunion with the owner dogs were focused mainly on the owner and on the environment, exhibiting safe base effect. A different approach was used to assess the welfare of shelter dogs. Dogs were implanted with a telemeter and after implantation dogs were housed in sequence in four different situations lasting 1 week: alone, alone with toys and a stretch cot for sleeping, with an unknown, spayed, female, and alone with a daily 2-hours interaction with an experimenter. Two different approaches were tried: partially random extracted fragments from every week, behaviors from 8 a.m. to 4 p.m. were continuous during baseline and the female situation. Results showed different reactions by dogs to the different situations and interestingly not all enrichments were enjoyed by the dogs improving their welfare. Overall results suggest that IRT may represent a useful tool to investigate emotional reactions in dogs. Nevertheless, further research is needed to establish the specificity and sensivity of IRT in this context and to assess how different dogs’ characteristics, breed, previous experience and the valence and arousal elicited by the stimulus could influence the magnitude and type of the response. The role of HRV in understanding emotional valence and the one of telemeters in understanding long-term effects on sheltered dogs’ welfare is also discussed.

The expression of the emotions in man and animals /

"Authorized edition."

Subjective Ambivalence and the Expression of Anti-Gay Bias

Univalent attitudes toward gay people have been widely studied, but no research to date has examined ambivalent (i.e., torn, conflicted) attitudes toward gay people. However, the Justification-Suppression Model (JSM; Crandall & Eshleman, 2003) proposes that ambivalence leads to biased expressions through intrapsychic processes which facilitate biased expression, particularly in contexts presenting strong justifications for expressing prejudice and weak pressures to suppress prejudice. I test these implications in the context of bias toward gay people. In Study 1, the measurement of ambivalence is examined in terms of both subjective ambivalence (i.e., the reported experience of “torn” attitudes) and calculated ambivalence (i.e., mathematical conflict between positive and negative attitude components). I find that higher subjective ambivalence is only associated with more negative attitudes toward gay people (and not positive attitudes toward gay people), and that higher subjective ambivalence predicts less gay rights support even after taking negative and positive attitudes toward gay people into account. Further, higher subjective ambivalence is associated with ideological opposition to gay people and more negative intergroup emotions (e.g., intergroup disgust). These findings suggest it is valuable to examine the unique component of subjective ambivalence separate from univalent negativity. Because calculated ambivalence measures are mathematically dependent upon a univalent negative measure, they cannot be examined separately from negativity. Therefore, subjective ambivalence is the focus of Study 2. The main goals of Study 2 were to determine why and when subjective ambivalence is related to bias. I examined the extent to which the negative relation between subjective ambivalence and opposition to anti-gay bullying can be accounted for by lower intergroup empathy and lower collective guilt, which may facilitate the expression of bias in keeping with the JSM. The relation between subjective ambivalence and anti-gay bullying opposition was examined within four social contexts based on a 2 (high vs. low offensiveness) x 2 (normatively unjustified vs. normatively justified) manipulation. I expected that higher subjective ambivalence would be most strongly related to lower intergroup empathy and collective guilt when there are the strongest justifications for bias expression, and that lower intergroup empathy and collective guilt would lead to less opposition to anti-gay bullying. Higher subjective ambivalence predicted less anti-gay bullying opposition. After accounting for positivity and negativity, the direct effect of subjective ambivalence was no longer significant, yet subjective ambivalence uniquely predicted intergroup empathy, which in turn predicted less anti-gay bullying opposition. These findings provide evidence that subjective ambivalence is largely negative in nature, but also presents evidence for a unique component of subjective ambivalence (separate from univalent attitudes) associated with low intergroup empathy and negativity. In contrast to previous research, I found very little evidence for the context-dependency of subjective ambivalence. Further research on subjective ambivalence, including subjective ambivalence toward other social groups, may expand our understanding of the factors leading to biased expressions.

The Influence of Parental Socialization of Emotions on Emotional Experience in Adulthood: An Exploratory Study.

Essai doctoral présenté à la Faculté des arts et des sciences en vue de l’obtention du grade de doctorat en psychologie clinique (D.Psy.)

A Cognitive Science Reasoning in Recognition of Emotions in Audio-Visual Speech

In this report we summarize the state-of-the-art of speech emotion recognition from the signal processing point of view. On the bases of multi-corporal experiments with machine-learning classifiers, the observation is made that existing approaches for supervised machine learning lead to database dependent classifiers which can not be applied for multi-language speech emotion recognition without additional training because they discriminate the emotion classes following the used training language. As there are experimental results showing that Humans can perform language independent categorisation, we made a parallel between machine recognition and the cognitive process and tried to discover the sources of these divergent results. The analysis suggests that the main difference is that the speech perception allows extraction of language independent features although language dependent features are incorporated in all levels of the speech signal and play as a strong discriminative function in human perception. Based on several results in related domains, we have suggested that in addition, the cognitive process of emotion-recognition is based on categorisation, assisted by some hierarchical structure of the emotional categories, existing in the cognitive space of all humans. We propose a strategy for developing language independent machine emotion recognition, related to the identification of language independent speech features and the use of additional information from visual (expression) features.

Pipkiiα Is Widely Expressed In Hematopoietic-derived Cells And May Play A Role In The Expression Of Alpha- And Gamma-globins In K562 Cells.

Characterized for the first time in erythrocytes, phosphatidylinositol phosphate kinases (PIP kinases) belong to a family of enzymes that generate various lipid messengers and participate in several cellular processes, including gene expression regulation. Recently, the PIPKIIα gene was found to be differentially expressed in reticulocytes from two siblings with hemoglobin H disease, suggesting a possible relationship between PIPKIIα and the production of globins. Here, we investigated PIPKIIα gene and protein expression and protein localization in hematopoietic-derived cells during their differentiation, and the effects of PIPKIIα silencing on K562 cells. PIPKIIα silencing resulted in an increase in α and γ globins and a decrease in the proliferation of K562 cells without affecting cell cycle progression and apoptosis. In conclusion, using a cell line model, we showed that PIPKIIα is widely expressed in hematopoietic-derived cells, is localized in their cytoplasm and nucleus, and is upregulated during erythroid differentiation. We also showed that PIPKIIα silencing can induce α and γ globin expression and decrease cell proliferation in K562 cells.

Diet Carotenoid Lutein Modulates The Expression Of Genes Related To Oxygen Transporters And Decreases Dna Damage And Oxidative Stress In Mice.

Lutein (LT) is a carotenoid obtained by diet and despite its antioxidant activity had been biochemically reported, few studies are available concerning its influence on the expression of antioxidant genes. The expression of 84 genes implicated in antioxidant defense was quantified using quantitative reverse transcription polymerase chain reaction array. DNA damage was measured by comet assay and glutathione (GSH) and thiobarbituric acid reactive substances (TBARS) were quantified as biochemical parameters of oxidative stress in mouse kidney and liver. cDDP treatment reduced concentration of GSH and increased TBARS, parameters that were ameliorated in treatment associated with LT. cDDP altered the expression of 32 genes, increasing the expression of GPx2, APC, Nqo1 and CCs. LT changed the expression of 37 genes with an induction of 13 mainly oxygen transporters. In treatments associating cDDP and LT, 30 genes had their expression changed with a increase of the same genes of the cDDP treatment alone. These results suggest that LT might act scavenging reactive species and also inducing the expression of genes related to a better antioxidant response, highlighting the improvement of oxygen transport. This improved redox state of the cell through LT treatment could be related to the antigenotoxic and antioxidant effects observed.

Quantitative Proteomic Analysis Reveals Metabolic Alterations, Calcium Dysregulation, And Increased Expression Of Extracellular Matrix Proteins In Laminin α2 Chain-deficient Muscle.

Congenital muscular dystrophy with laminin α2 chain deficiency (MDC1A) is one of the most severe forms of muscular disease and is characterized by severe muscle weakness and delayed motor milestones. The genetic basis of MDC1A is well known, yet the secondary mechanisms ultimately leading to muscle degeneration and subsequent connective tissue infiltration are not fully understood. In order to obtain new insights into the molecular mechanisms underlying MDC1A, we performed a comparative proteomic analysis of affected muscles (diaphragm and gastrocnemius) from laminin α2 chain-deficient dy(3K)/dy(3K) mice, using multidimensional protein identification technology combined with tandem mass tags. Out of the approximately 700 identified proteins, 113 and 101 proteins, respectively, were differentially expressed in the diseased gastrocnemius and diaphragm muscles compared with normal muscles. A large portion of these proteins are involved in different metabolic processes, bind calcium, or are expressed in the extracellular matrix. Our findings suggest that metabolic alterations and calcium dysregulation could be novel mechanisms that underlie MDC1A and might be targets that should be explored for therapy. Also, detailed knowledge of the composition of fibrotic tissue, rich in extracellular matrix proteins, in laminin α2 chain-deficient muscle might help in the design of future anti-fibrotic treatments. All MS data have been deposited in the ProteomeXchange with identifier PXD000978 (http://proteomecentral.proteomexchange.org/dataset/PXD000978).

Histologic Correlation Of Expression Of Ki-67 In Squamous Cell Carcinoma Of The Glottis According To The Degree Of Cell Differentiation.

Squamous cell carcinoma is the most common neoplasm of the larynx and glottis, and its prognosis depends on the size of the lesion, level of local invasion, cervical lymphatic spread, and presence of distant metastases. Ki-67 (MKI67) is a protein present in the core, whose function is related to cell proliferation. To evaluate the expression of marker Ki-67 in squamous cell carcinoma of the larynx and glottis and its correlation to pathological findings. Experimental study with immunohistochemistry analysis of Ki-67, calculating the percentage of the cell proliferation index in glottic squamous cell carcinomas. Sixteen cases were analyzed, with six well-differentiated and 10 poorly/moderately differentiated tumors. There was a correlation between cell proliferation index and degree of cell differentiation, with higher proliferation in poorly/moderately differentiated tumors. The cell proliferation index, as measured by Ki-67, may be useful in the characterization of histological degree in glottic squamous cell tumors.

Decreased Expression Of Stem Cell Markers By Simvastatin In 7,12-dimethylbenz(a)anthracene (dmba)-induced Breast Cancer.

Simvastatin, a competitive inhibitor of HMG-CoA reductase widely used in the treatment and prevention of hyperlipidemia-related diseases, has recently been associated to in vitro anticancer stem cell (CSC) actions. However, these effects have not been confirmed in vivo. To assess in vivo anti-CSC effects of simvastatin, female Sprague-Dawley rats with 7,12-dimethyl-benz(a)anthracene (DMBA)-induced mammary cancer and control animals were treated for 14 days with either simvastatin (20 or 40 mg/kg/day) or soybean oil (N = 60). Tumors and normal breast tissues were removed for pathologic examination and immunodetection of CSC markers. At 40 mg/kg/day, simvastatin significantly reduced tumor growth and the expression of most CSC markers. The reduction in tumor growth (80%) could not be explained solely by the decrease in CSCs, since the latter accounted for less than 10% of the neoplasia (differentiated cancer cells were also affected). Stem cells in normal, nonneoplastic breast tissues were not affected by simvastatin. Simvastatin was also associated with a significant decrease in proliferative activity but no increase in cell death. In conclusion, this is the first study to confirm simvastatin anti-CSC actions in vivo, further demonstrating that this effect is specific for neoplastic cells, but not restricted to CSCs, and most likely due to inhibition of cell proliferation.

A Leucine-rich Diet Modulates The Tumor-induced Down-regulation Of The Mapk/erk And Pi3k/akt/mtor Signaling Pathways And Maintains The Expression Of The Ubiquitin-proteasome Pathway In The Placental Tissue Of Nmri Mice.

Placental tissue injury is concomitant with tumor development. We investigated tumor-driven placental damage by tracing certain steps of the protein synthesis and degradation pathways under leucine-rich diet supplementation in MAC16 tumor-bearing mice. Cell signaling and ubiquitin-proteasome pathways were assessed in the placental tissues of pregnant mice, which were distributed into three groups on a control diet (pregnant control, tumor-bearing pregnant, and pregnant injected with MAC-ascitic fluid) and three other groups on a leucine-rich diet (pregnant, tumor-bearing pregnant, and pregnant injected with MAC-ascitic fluid). MAC tumor growth down-regulated the cell-signaling pathways of the placental tissue and decreased the levels of IRS-1, Akt/PKB, Erk/MAPK, mTOR, p70S6K, STAT3, and STAT6 phosphorylated proteins, as assessed by the multiplex Millipore Luminex assay. Leucine supplementation maintained the levels of these proteins within the established cell-signaling pathways. In the tumor-bearing group (MAC) only, the placental tissue showed increased PC5 mRNA expression, as assessed by quantitative RT-PCR, decreased 19S and 20S protein expression, as assessed by Western blot analysis, and decreased placental tyrosine levels, likely reflecting up-regulation of the ubiquitin-proteasome pathway. Similar effects were found in the pregnant injected with MAC-ascitic fluid group, confirming that the effects of the tumor were mimicked by MAC-ascitic fluid injection. Although tumor progression occurred, the degradation pathway-related protein levels were modulated under leucine-supplementation conditions. In conclusion, tumor evolution reduced the protein expression of the cell-signaling pathway associated with elevated protein degradation, thereby jeopardizing placental activity. Under the leucine-rich diet, the impact of cancer on placental function could be minimized by improving the cell-signaling activity and reducing the proteolytic process.

Increased Expression Of Hes5 Protein In Notch Signaling Pathway In The Hippocampus Of Mice Offspring Of Dams Fed A High-fat Diet During Pregnancy And Suckling.

Maternal high-fat diet (HFD) impairs hippocampal development of offspring promoting decreased proliferation of neural progenitors, in neuronal differentiation, in dendritic spine density and synaptic plasticity reducing neurogenic capacity. Notch signaling pathway participates in molecular mechanisms of the neurogenesis. The activation of Notch signaling leads to the upregulation of Hes5, which inhibits the proliferation and differentiation of neural progenitors. This study aimed to investigate the Notch/Hes pathway activation in the hippocampus of the offspring of dams fed an HFD. Female Swiss mice were fed a control diet (CD) and an HFD from pre-mating until suckling. The bodyweight and mass of adipose tissue in the mothers and pups were also measured. The mRNA and protein expression of Notch1, Hes5, Mash1, and Delta1 in the hippocampus was assessed by RT-PCR and western blotting, respectively. Dams fed the HFD and their pups had an increased bodyweight and amount of adipose tissue. Furthermore, the offspring of mothers fed the HFD exhibited an increased Hes5 expression in the hippocampus compared with CD offspring. In addition, HFD offspring also expressed increased amounts of Notch1 and Hes5 mRNA, whereas Mash1 expression was decreased. However, the expression of Delta1 did not change significantly. We propose that the overexpression of Hes5, a Notch effector, downregulates the expression of the proneural gene Mash1 in the offspring of obese mothers, delaying cellular differentiation. These results provide further evidence that an offspring's hippocampus is molecularly susceptible to maternal HFD and suggest that Notch1 signaling in this brain region is important for neuronal differentiation.

The recurrence of leprosy reactional episodes could be associated with oral chronic infections and expression of serum IL-1, TNF-α, IL-6, IFN-γ and IL-10

The aim of this study was to determine whether the presence of leprosy reactional episodes could be associated with chronic oral infection. Thirty-eight leprosy patients were selected and divided into 2 groups: group I - 19 leprosy patients with oral infections, and group II - 19 leprosy patients without oral infections. Ten patients without leprosy, but presenting oral infections, were assigned to the control group. Leprosy patients were classified according to Ridley and Jopling classification and reactional episodes of the erythema nodosum type or reversal reaction were identified by clinical and histopathological features associated with serum IL-1, TNF-α, IL-6, IFN-γ and IL-10 levels. These analyses were performed immediately before and 7 days after the oral infection elimination. Patients from group I presenting oral infections reported clinical improvement of the symptoms of reactional episodes after dental treatment. Serum IL-1, TNF-α, IL-6, IFN-γ and IL-10 levels did not differ significantly before and after dental treatment as determined by the Wilcoxon test (p>0.05). Comparison of the 2 groups showed statistically significant differences in IL-1 and IL-6 at baseline and in IL-1, IL-6 and IL-10 on the occasion of both collections 7 days after therapy. Serum IL-6 and IL-10 levels in group I differed significantly at baseline compared to control (Mann-Whitney test; p<0.05). These results suggest that oral infection could be involved as a maintenance factor in the pathogenesis of leprosy reactional episodes.

Immunohistochemical expression of p53, p16 and hTERT in oral squamous cell carcinoma and potentially malignant disorders

Oral carcinogenesis is a multi-step process. One possible step is the development of potentially malignant disorders known as leukoplakia and erytroplakia. The objective of this study was to use immunohistochemistry to analyze the patterns of expression of the cell-cycle regulatory proteins p53 and p16INK4a in potentially malignant disorders (PMD) of the oral mucosa (with varying degrees of dysplasia) and in oral squamous cell carcinomas (OSCC) to correlate them with the expression of telomerase (hTERT). Fifteen PMD and 30 OSCC tissue samples were analyzed. Additionally, 5 cases of oral epithelial hyperplasia (OEH) were added to analyze clinically altered mucosa presenting as histological hyperplasia without dysplasia. p53 positivity was observed in 93.3% of PMD, in 63.3% of OSCC and in 80% of OEH. Although there was no correlation between p53 expression and the grade of dysplasia, all cases with severe dysplasia presented p53 suprabasal immunoexpression. p16INK4a expression was observed in 26.7% of PMD, in 43.3% of OSCC and in 2 cases of OEH. The p16INK4a expression in OEH, PMD and OSCC was unable to differentiate non-dysplastic from dysplastic oral epithelium. hTERT positivity was observed in all samples of OEH and PMD and in 90% of OSCC. The high hTERT immunoexpression in all three lesions indicates that telomerase is present in clinically altered oral mucosa but does not differentiate hyperplastic from dysplastic oral epithelium. In PMD of the oral mucosa, the p53 immunoexpression changes according to the degree of dysplasia by mechanisms independent of p16INK4a and hTERT.