Subchronic oral toxicity study of furan in B6C3F1 Mice.

Furan is a heterocyclic organic compound formed during heat treatment for processing and preservation of various types of food. Rodent studies have previously shown that furan is a hepatocarcinogen. Those studies were conducted over a high dose range, which induced tumors at nearly 100% incidence at all doses. This ninety-day gavage study in mice was conducted to extend the dose to a lower range (0.0, 0.03, 0.12, 0.5, 2.0, and 8.0 mg/kg body weight [bw] per day) to identify a no-observed adverse effect level for hepatotoxicity and to characterize non-neoplastic effects, including those affecting clinical biochemistry, hematology, tissue morphology, and histopathology. The liver was the primary target organ with dose-dependent toxicity. Liver weights were increased at the 8.0 mg/kg bw dose in females only. Levels of the serum enzyme alanine transaminase, representative of liver damage, were increased three-fold at the highest dose. Histological changes in the liver were observed at 2.0 and 8.0 mg/kg bw in both sexes. Although clinical parameters were also altered for the kidney, these differences were not accompanied by histological changes. Based on these clinical biochemical and histological changes, a no-observed adverse effect level of 0.12 mg/kg bw per day of furan in mice is suggested.

Subchronic oral toxicity study of furan in Fischer-344 rats.

Rodent studies have shown that furan is a hepatocarcinogen. Previous studies conducted with high doses showed tumors at nearly 100% incidence at all doses. In this paper, a ninety-day gavage experiment conducted with lower doses (0.0, 0.03, 0.12, 0.5, 2.0, and 8.0 mg/kg bw) to identify a no-observed adverse effect level for hepatotoxicity and to characterize non-neoplastic effects including gross changes and histopathology, clinical biochemistry, hematology, and immunotoxicology is reported. As indicated by changes in serum biomarkers, increased liver weights and gross and histological lesions, the liver is the major target organ affected by furan. There were no changes in body weights, food consumption, or histology in other organs. Some of the serum electrolyte markers, including phosphorus, were altered. There was a significant increase in serum thyroxine and triidothyronine in males. This increase was not accompanied by histological thyroid changes. Immunophenotypic analysis showed that thymic lymphocyte maturation was altered in male rats. Although altered clinical biochemistry and hematological parameters were observed at a dose of > 0.5 mg/kg bw, mild histological lesions in the liver were observed at > 0.12 mg/kg bw. Based on this finding, a furan dose of 0.03 mg/kg bw was proposed as the no-observed adverse effect level for hepatic toxicity.

The distribution of tachykinin binding sites in the brain of an electric fish (Apteronotus leptorhynchus).

We mapped the distribution of tachykinin binding sites utilizing quantitative autoradiography of iodinated substance P and eledoisin as prototypic ligands for neurokinin-1 (NK1) and neurokinin-3 (NK3) receptors, respectively. The two ligands produced highly heterogenous and quantitatively different patterns of specific binding, suggesting that they revealed different tachykinin receptor subtypes. Although [125I]substance P and [125I]eledoisin binding were correlated in most brain regions, the binding of substance P was usually denser. [125I]substance P binding and substance P-like immunoreactivity were reasonably correlated in most brain areas, although discrepancies were found in some nuclei. Dense [125I]substance P binding was found in most areas of the subpallium and in parts of the pallium related to the olfactory system, as well as in the glomerular layer of the olfactory bulb. Moderate to dense binding of both ligands was observed in preoptic area, hypothalamus, habenula, parts of the thalamus and preglomerular complex. Especially noteworthy was the presence of [125I] substance P binding in the diencephalic prepacemaker nucleus, a region involved in the control of electroncommuncatory behavior. Substance P-like immunoreactivity is sexually dimorphic in certain diencephalic nuclei, including the prepacemaker nucleus (Weld and Maler, 1992); no obvious difference was seen between [125I]substance P or [125I]eledoisin binding in the brains of male versus female fish. In the mesencephalon striking laminar patterns of binding were seen in the torus semicircularis dorsalis and the optic tectum. Dense binding was also noted in the raphé nuclei, the locus ceruleus and the sensory nucleus of the vagus. Although binding of substance P in the electrosensory lateral line lobe and nucleus preeminentialis was light, it was distributed in a discrete fashion, suggesting a role of substance P in electrosensory processing.

Connections of the olfactory bulb in the gymnotiform fish, Apteronotus leptorhynchus.

This work examines the connectivity of the olfactory bulb in the gynmotiform fish Apteronotus leptorhynchus. Wheat germ agglutinin conjugated horseradish peroxidase was iontophoresed in different areas and depths of the bulb in order to define its efferent and afferent connections. The olfactory bulb projects bilaterally via the medial (medial and centromedial fascicles) and lateral olfactory (lateral and centrolateral fascicles) tracts. The nervus terminalis courses through the ventromedial aspect of the bulb to terminate in parts of the medial subpallium and hypothalamus. Its telencephalic component could be identified by a nonpreadsorbable substance P-like immunoreactivity. Fibers within the medial olfactory tract form four telencephalic terminal fields: peduncular, medial, intermediate and posterior fields. The diencephalic terminal fields in the habenula, preoptic, and hypothalamic areas appear to correspond to some of the nervus terminalis fibers (von Bartheld and Meyer [1986] Cell Tissue Res. 245:143-158, Krishna et al. [1992] Gen. Comp. Endocrinol. 85:111-117), and to axons of telencephalic bulbopetal cells of area dorsalis posterior. The terminal fields of the medial olfactory tract and nervus terminalis partially overlap in the ventral telencephalic areas partes ventralis, supracommissuralis, and rostral preoptic region. The lateral olfactory tract forms a lateral terminal field and contributes to the intermediate and posterior terminal fields. Olfactory fibers cross in the interbulbar, anterior, and habenular commissures and tuberal decussation. Consistent differences were noted between the medial and lateral olfactory bulb, with respect to their cytoarchitectonics, immunohistochemistry, and connections. In addition to the olfactory nerve, bulbar afferents are predominantly ipsilateral, with minor inputs originating from the contralateral bulb and telencephalic area dorsalis posterior, nucleus raphe centralis, and locus ceruleus.

Substance P-like immunoreactivity in the brain of the gymnotiform fish Apteronotus leptorhynchus: presence of sex differences.

The distribution of substance P-like immunoreactivity (SPli) was charted in the brain of the gymnotiform fish Apteronotus leptorhynchus, and correlated with the circuitry underlying intraspecific electrocommunication. Cell bodies were found predominantly in the lateral hypothalamus and in certain paraventricular organs: nucleus preopticus periventricularis, anterior subdivision; anterior hypothalamus; nucleus posterioris periventricularis; nucleus recessus lateralis, medial subdivision 2; nucleus recessus posterioris and nucleus recessus lateralis, lateral subdivision. Cell bodies were also found in the rostral olfactory nucleus, ventral telencephalon (ventral and central subdivisions), the habenula, the vagal sensory and motor nuclei and in the subtrigeminal nucleus. The distribution of SPli fibers was similar in some respects to that reported for other vertebrates. SPli was found in the rhombencephalon associated with vagal afferent fibers and in the funicular nucleus (possibly related to nociception). In the diencephalon and midbrain SPli fibers were found in the habenular-interpeduncular tract, in the hypothalamus and pituitary. SPli fibers were also found in preoptic and forebrain areas. The most striking result was the sexually dimorphic SPli innervation of certain hypothalamic and septal nuclei, and of the prepacemaker nucleus (PPn), a diencephalic cell group which controls communication ('chirping') in gymnotiforms. The PPn and septal/hypothalamic nuclei were densely innervated by SPli in males but devoid of SPli in females.

Angiotensin II stimulation of teleost adrenocorticotropic hormone release: interactions with urotensin I and corticotropin-releasing factor.

The release of adrenocorticotropic hormone (ACTH) from dispersed goldfish anterior pituitary cells was examined in order to determine whether human angiotensin II (AII) would potentiate the ACTH-releasing activity of urotensin I (UI) and ovine corticotropin-releasing factor (oCRF), peptides with a sequence homology of greater than 50%. In mammals, AII has a slightly enhancing or potentiating effect on CRF-stimulated ACTH release. In the present investigations, concentrations of AII (0.5 and 1 nM), and of UI (1 nM) or oCRF (3 nM), which elicit moderate increases in ACTH release, were tested alone and in combination. The ACTH-releasing activities of AII and UI combined, or of AII and oCRF combined, showed no potentiation and in fact were less than additive. It was concluded that AII does not potentiate the ACTH-releasing activity of either UI or oCRF observed with goldfish anterior pituitary cells in vitro.

Inhibition of CRF- and urotensin I-stimulated ACTH release from goldfish pituitary cell columns by the CRF analogue alpha-helical CRF-(9-41).

alpha-Helical CRF-(9-41) is an analogue of corticotropin-releasing factor (CRF) that antagonizes CRF-stimulated ACTH release in rats. In the present study, alpha-helical CRF-(9-41) was tested to determine whether it would antagonize the ACTH-releasing activity of CRF or urotensin I (UI) observed with superfused, dispersed goldfish anterior pituitary cells. At a concentration of 4 microM, alpha-helical CRF-(9-41) completely blocked the ACTH-releasing activity of 100 nM CRF or 100 nM UI. The inhibitor by itself showed little intrinsic ACTH-releasing activity. This investigation reveals similarities in the CRF-antagonism of alpha-helical CRF-(9-41) in the teleost and mammalian pituitary in vitro. It also provides are similar and suggests that alpha-helical CRF-(9-41) may be useful as a tool to investigate the effects of CRF-like and UI-like peptides in teleost fishes.

Stimulation by angiotensins I and II of ACTH release from goldfish pituitary cell columns.

Dispersed, superfused goldfish anterior pituitary cell columns were stimulated with pulses of salmon angiotensin I (sAI), human angiotensin I (hAI), and human angiotensin II (hAII). Human AII stimulated the greatest release of ACTH. The dose-response curves for hAI and sAI were similar and revealed that hAI and sAI were about one-tenth as potent as hAII in stimulating ACTH release. In mammals, AI must be converted to AII in order to stimulate ACTH release. In goldfish, the angiotensin-converting enzyme inhibitor captopril, which inhibits the conversion of AI to AII, was not able to block sAI-stimulated ACTH release. This finding suggests that the angiotensin receptor of the goldfish corticotrope is less discriminating than that of the mammalian corticotrope and recognizes both AI and AII. This hypothesis was supported by the observation that sarcosine analogs of AII, which block AII-stimulated ACTH release in mammals, failed to block hAII-stimulated ACTH release in goldfish. Saralasin showed negligible, [Sar1,Thr8]-AII slight, and [Sar1, Ile8]-AII moderate, intrinsic ACTH-releasing activity. These findings suggest that the ACTH-releasing activity of angiotensin appeared early in the evolution of the vertebrate pituitary.

Circadian and seasonal variations of lipogenesis in the Gulf killifish, Fundulus grandis.

Lipogenesis was measured in male Gulf killifish at three seasons under different environmental conditions. Cold temperature is stimulatory for lipogenesis in the fall and spring, but warm temperature is more stimulatory during the summer. The highest rate of lipogenesis occurs during the spring. However, total body fat content is relatively low at that time, which suggests that lipids are being actively mobilized as well. A high rate of lipogenesis also occurs at a cold temperature (20 degrees C) during the fall, and more lipids are stored at that time of year, as evidenced by higher liver and body fat stores. Both lipogenesis and total body fat content are at a minimum during the summer. These results indicate that high levels of lipid synthesis do not necessarily result in higher body fat content, as the latter is the product of both lipid synthesis and lipid mobilization. This study is also in agreement with previous studies which had indicated a changing responsiveness to temperature during the annual cycle of the Gulf killfish.