Daniela Cevolani

Results of Long‐Term Experimental Studies on the Carcinogenicity of Methyl Alcohol and Ethyl Alcohol in Rats

Abstract: Methyl alcohol was administered in drinking water supplied ad libitum at doses of 20,000, 5,000, 500, or 0 ppm to groups of male and female Sprague‐Dawley rats 8 weeks old at the start of the experiment. Animals were kept under observation until spontaneous death. Ethyl alcohol was administered by ingestion in drinking water at a concentration of 10% or 0% supplied ad libitum to groups of male and female Sprague‐Dawley rats; breeders and offspring were included in the experiment. Treatment started at 39 weeks of age (breeders), 7 days before mating, or from embryo life (offspring) and lasted until their spontaneous death. Under tested experimental conditions, methyl alcohol and ethyl alcohol were demonstrated to be carcinogenic for various organs and tissues. They must also be considered multipotential carcinogenic agents. In addition to causing other tumors, ethyl alcohol induced malignant tumors of the oral cavity, tongue, and lips. These sites have been shown to be target organs in man by epidemiologic studies.

Results of Long‐Term Experimental Studies on the Carcinogenicity of Ethylene‐bis‐Dithiocarbamate (Mancozeb) in Rats

Abstract: Mancozeb, an ethylene‐bis‐dithiocarbamate (EBDC), has been one of the most commonly used fungicides in commercial use for several decades. Nevertheless, up to now, no adequate published experimental studies on the carcinogenicity of Mancozeb have been published. Because of the importance of the compound and of the number of people potentially exposed (workers engaged in the production and use of the fungicide, people living in agricultural areas where the compound is sprayed, and people consuming polluted products), a long‐term experimental study of Mancozeb was begun at the Cancer Research Center of the Ramazzini Foundation. Groups of 150 male and female Sprague‐Dawley rats, 8 weeks old at the start of the treatment, were administered Mancozeb at the concentration of 1000, 500, 100, 10, and 0 ppm in feed supplied ad libitum for 104 weeks. At the end of the treatment, animals were kept under controlled conditions until spontaneous death. Mancozeb caused an increase in (1) total malignant tumors, (2) malignant mammary tumors, (3) Zymbal gland and ear duct carcinomas, (4) hepatocarcinomas, (5) malignant tumors of the pancreas, (6) malignant tumors of the thyroid gland, (7) osteosarcomas of the bones of the head, and (8) hemolymphoreticular neoplasias. On the basis of these data, Mancozeb must be considered a multipotent carcinogenic agent.

Thermosensitivity of anterior hypothalamic-preoptic neurons during the waking-sleeping cycle: a study in brain functional states

The thermosensitivity of anterior hypothalamic-preoptic neurons was studied in cats during the waking-sleeping cycle. Direct cooling and warming of the anterior hypothalamic-preoptic region was accomplished with water-perfused thermodes. Neuronal thermosensitivity was determined by means of the linear regression analysis of firing rate changes vs anterior hypothalamic-preoptic temperature changes. A total of 117 neurons were classified as thermosensitive during wakefulness and synchronized sleep (20.1% of the studied neurons). Cold-sensitive neurons outnumbered warm-sensitive neurons by 3.7:1. The homeothermic states, wakefulness and synchronized sleep, are characterized by similar frequency distributions of neuronal thermosensitivity, although variable changes in single neuron thermosensitivity are state-dependent. Such changes underlie the quantitative differences in homeothermic regulation between these states. The impairment of thermoregulation during desynchronized sleep is characterized by a different frequency distribution of neuronal thermosensitivity resulting from both a drop in the responsiveness to thermal stimulation of a majority of neurons and a reversal in the sensitivity to cooling and warming of a minority of neurons. In conclusion, only the frequency distribution of thermosensitivity in the neuronal population is indicative of changes in the thermoregulation paradigm across behavioral states.

Responses of anterior hypothalamic-preoptic neurons to direct thermal stimulation during wakefulness and sleep

The responses of anterior hypothalamic-preoptic units to direct thermal stimulation were studied during wakefulness and sleep in cats. Seventy-nine percent of the selected units showed changes in firing rate in relation to wakefulness and sleep stages. Forty-nine percent of the units characterized by activity related to EEG patterns were found to be responsive to thermal stimulation in wakefulness and synchronized sleep. Unit responses to thermal stimulation were either absent or inconsistent in desynchronized sleep.

Brain functional changes in patients with ulcerative colitis: A functional magnetic resonance imaging study on emotional processing

Background: Ulcerative colitis (UC) is associated with psychological stress and poor emotional functioning. The neural emotional processing involves the complex integration of several cortical and subcortical brain structures. The amygdala plays a fundamental role in the neural processing of emotional stimuli and is a core structure of the brain–gut axis (BGA) that represents the anatomo‐functional substrate for the bidirectional influences between emotions and gastrointestinal functions. The aim of this study was to investigate the brain emotional processing in UC patients compared to healthy people. Methods: Ten UC patients in remission and 10 matched healthy controls underwent a functional magnetic resonance imaging (fMRI) scan while performing a task involving emotional visual stimuli. A set of negative, positive, and neutral pictures were used to study brain‐related emotional responses. Results: A significantly reduced blood oxygen level‐dependent (BOLD) signal in UC patients relative to controls was found in the amygdala, thalamic regions, and cerebellar areas (P < 0.05 corrected for multiple comparisons). The group‐related differences were detected in the brain activity in response to positive emotional stimuli. Conclusions: UC is associated with an emotional dysfunction characterized by decreased sensitivity to emotions with a positive content. The previous intestinal inflammatory activity in UC patients might have contributed to determine the functional changes of the amygdala that we found. On the other hand, the dysfunction of the amygdala may influence the course of the disease. (Inflamm Bowel Dis 2010;)

Amygdala responses to masked and low spatial frequency fearful faces: a preliminary fMRI study in panic disorder

Previous studies have demonstrated amygdala activation in response to fearful faces even if presented below the threshold of conscious visual perception. It has also been proposed that subcortical regions are selectively sensitive to low spatial frequency (LSF) information. However, chronic hyperarousal may reduce amygdala activation in panic disorder (PD). Our aim was to establish whether the amygdala is engaged by masked and LSF fearful faces in PD as compared to healthy subjects. Neutral faces were used as the mask stimulus. Thirteen PD patients (seven females, six males; mean age=29.1 (S.D: 5.9)) and 15 healthy volunteers (seven females, eight males; mean age=27.9 (S.D. 4.5)) underwent two passive viewing tasks during a 3T functional magnetic resonance imaging (fMRI) as follows: 1) presentation of faces with fearful versus neutral expressions (17ms) using a backward masking procedure and 2) presentation of the same faces whose spatial frequency contents had been manipulated by low-pass filtering. Level of awareness was confirmed by a forced choice fear-detection task. Whereas controls showed bilateral activation to fearful masked faces versus neutral faces, patients failed to show activation within the amygdala. LSF stimuli did not elicit amygdala response in either group, contrary to the view that LSF information plays a crucial role in the processing of facial expressions in the amygdala. Findings suggest maladaptive amygdala responses to potentially threatening visual stimuli in PD patients.

Polygraphic study of anterior hypothalamic-preoptic neuron thermosensitivity during sleep ☆

Thermal responsiveness of anterior hypothalamic-preoptic (AH-PO) neurons was studied in unanesthetized sleeping cats carrying chronically implanted electrodes and thermistors for recording EEG, EMG and AH-PO temperature. Activity of AH-PO neurons was recorded using tungsten microelectrodes during atraumatic stereotaxic head restraint in a sound-attenuated chamber at 22-25 degrees C. Direct cooling or warning of the AH-PO region was accomplished with bilaterally positioned water-perfused thermodes. 110 thermosensitive neurons (77% cold-sensitive and 23% warm-sensitive) were sampled during wakefulness and synchronized sleep. In contrast, the changes in spontaneous firing rate shown by AH-PO neurons during desynchronized sleep were often associated with a strong depression or outright suppression of their responsiveness to AH-PO thermal stimulation.

Different localization of spermidine/spermine N1-acetyltransferase and ornithine decarboxylase transcripts in the rat kidney.

In situ hybridization histochemistry of transverse sections from male rat kidney showed that the mRNA of the regulatory enzyme of polyamine degradation, spermidine/spermine N1 acetyltransferase, has a spotty distribution in the cortex, is low and diffused in the outer stripe and high and diffused in the inner stripe of the outer medulla. At the cellular level, this mRNA is solely expressed by the epithelium of the distal straight and convoluted nephron tubules. Since biosynthetic ornithine decarboxylase mRNA is solely found in the proximal straight tubules, it is proposed that polyamine biosynthesis and degradation occur at separate sites along the nephron.

Neuronal loss up-regulates clusterin mRNA in living neurons and glial cells in the rat brain

The aim of this study was to examine the time course and cellular localization of clusterin mRNA after neurodegeneration. Selective neuronal death was achieved in the rat inferior olivary complex after volkensin injection in the contralateral cerebellar cortex. Quantitative analysis of the in situ hybridization signal demonstrated over-expression of clusterin mRNA in living neurons at 6 days and outside the neuronal cell bodies at 10 days post-injection. We conclude that, in our experimental model, clusterin over-expression occurs as an early and transient neuronal and as a delayed glial response to selective neuronal death, supporting the view that clusterin may be involved in cytoprotection and tissue remodeling.

Characterization and Regional Distribution of a Class of Synapses with Highly Concentrated cAMP Binding Sites in the Rat Brain

A class of putative synaptic terminals with concentrated cAMP binding sites are labelled in unfixed sections of rat brain by means of the ligand 8‐thioacetamido fluorescein cAMP (SAF‐cAMP), a fluorescent analogue of cAMP. The labelled terminals appear as sharply delimited bouton‐like structures in close proximity but external to the cell body of neurons. The SAF‐cAMP binding, measured at equilibrium in competition with other nucleotides, indicates that the binding site recognizes the cAMP moiety of SAF‐cAMP. In the labelled terminals of the frontal cortex the concentration of SAF‐cAMP binding sites is estimated to be in the millimolar range (at least 2.1 ± 1.0 mM). In a brain homogenate, labelled terminals are visualized only in the membrane fraction enriched in synaptosomes. The cAMP binding activity of the synaptosomes is insoluble in high and in low ionic strength solution and is only partially solubilized by detergents, suggesting that the binding sites are intrinsic membrane proteins and/or proteins associated with the cytoskeleton. There is the possibility that SAF‐cAMP labels new cAMP binding sites highly concentrated in a class of synaptic terminals. SAF‐cAMP labelling is prominent in well defined regions of the rat brain: (i) the frontal and entorhinal areas of the cortex; (ii) the field CA1 of the hippocampus; (iii) the olfactory system; (iv) the medial nuclei of the thalamus; (v) the parabrachial nuclei and other less defined regions of the reticular substance; (vi) the substantia gelatinosa of Rolando in the spinal cord; and (vii) the neo‐ and paleocerebellum in the Purkinje cell layer, the archicerebellum in the granular cell layer. SAF‐cAMP labelling is absent in specific motor and sensory structures, with the exception of the olfactory system. It is proposed that SAF‐cAMP binding sites single out a new type of synaptic terminals involved in complex nervous functions.