Immunohistochemical expression of p63, Ki67 and ß-catenin in canine transitional cell carcinoma and polypoid cystitis of the urinary bladder.

Transitional cell carcinoma (TCC) is a urinary bladder tumour associated with high mortality in dogs. In this study, we investigated the feasibility of using p63, Ki67 or ß-catenin as a clinical marker for predicting biological behaviour and prognosis in canine TCC. Expression levels of these proteins in TCC (n = 25), polypoid cystitis (n = 5) and normal urinary bladder (n = 5) were scored after immunohistochemical staining. The staining scores for p63 (P < 0.01) and ß-catenin (P < 0.05) in TCC were significantly lower than those in normal urinary bladder and polypoid cystitis. In contrast, Ki67 (P < 0.01) staining scores in TCC were significantly higher than those in normal urinary bladder and polypoid cystitis. In TCC, low p63 expression was significantly related to the presence of vessel invasion (P < 0.05) and metastasis (P < 0.01) as well as short survival time (P < 0.05). These findings show that p63 could be a reliable marker for predicting prognosis in canine TCC.

Hemokinin-1 mediates pruriceptive processing in the rat spinal cord.

Hemokinin-1 (HK-1) is a new mammalian tachykinin peptide consisting of the amino acid sequence similar to substance P (SP). Although the function of SP, a representative tachykinin peptide, has been well established in the pain system, that of HK-1 has not yet been elucidated. [Leu(11)]-SP had an antagonistic effect on SP-induced scratching behavior, suggesting that [Leu(11)]-HK-1 may also attenuate the induction of scratching behavior by HK-1. Thus, the effects of a pretreatment with [Leu(11)]-HK-1 were evaluated to clarify the function of HK-1. The intrathecal administration of [Leu(11)]-HK-1 attenuated the induction of scratching by HK-1, but not SP, while [Leu(11)]-SP reduced the induction of scratching by SP, but not HK-1. These results indicated that [Leu(11)]-HK-1 may be a more specific antagonist of HK-1-preferred receptors and [Leu(11)]-SP has an antagonistic effect on the SP-preferred receptor, the neurokinin 1 receptor. In the formalin test for examining noxious response, the intrathecal administration of [Leu(11)]-SP, but not [Leu(11)]-HK-1, reduced the number of flinchings and c-Fos-positive cells in the spinal dorsal horn following formalin injection into the plantar region of the hind paw. These results indicated that SP, but not HK-1, is involved in nociceptive processing at the spinal level. To evaluate the involvement of HK-1 and SP in pruritic processing, the effect of [Leu(11)]-HK-1 and [Leu(11)]-SP on the induction of scratching behavior and c-Fos expression by serotonin (5-HT) and histamine was evaluated. The increased induction of scratching behavior and c-Fos expression by 5-HT and histamine was markedly attenuated by pretreatment with both [Leu(11)]-HK-1 and [Leu(11)]-SP, suggesting that HK-1 and SP may be involved in pruritic processing. These results indicate that HK-1 is involved in pruritic processing and [Leu(11)]-HK-1 is a valuable tool for clarifying the mechanisms underlying pruritic processing.

An amino-terminal fragment of hemokinin-1 has an inhibitory effect on pruritic processing in rats.

Hemokinin-1 (HK-1) is a peptide encoded by the preprotachykinin gene, TAC-4, and shares the hydrophobic carboxyl-terminal (C-terminal) region common to mammalian tachykinin peptides, such as substance P (SP). It is generally believed that C-terminal fragments of SP elicit an excitatory effect, while pretreatment with amino-terminal (N-terminal) fragments of SP inhibits the function of SP; however, there is no available information on HK-1. Therefore, to clarify the characteristics of C-terminal and N-terminal fragments of HK-1, HK-1 was divided into HK-1 (1-5) as the N-terminal fragment and HK-1 (6-11) as the C-terminal fragment based on the similarity of amino acids between HK-1 and SP. Intrathecal administration of HK-1 (6-11) induced scratching behavior similar to HK-1, while HK-1 (1-5) hardly induced scratching. Pretreatment with HK-1 (1-5), however, attenuated scratching induced by HK-1 and SP, whereas pretreatment with SP (1-5) attenuated SP-induced scratching, but not HK-1. Furthermore, intrathecal administration of HK-1 (1-5) and SP (1-5) markedly attenuated the induction of flinching and enhancement of c-Fos expression in the spinal cord following the intradermal administration of formalin, a noxious stimulant, while pretreatment with HK-1 (1-5), but not SP (1-5), markedly attenuated the induction of scratching behavior by subcutaneous administration of pruritic agents, such as serotonin or histamine. Taken together, these findings indicate that HK-1 (1-5) suppresses pruritic and nociceptive processing, while SP (1-5) suppresses nociceptive processing. Therefore, it is suggested that HK-1 (1-5) may be a useful tool for revealing pruritic processing and HK-1 may play a crucial role in pruritic processing.

The amino-terminal region of hemokinin-1 regulates the induction of thermal hyperalgesia in rats.

It is known that intrathecal administration of substance P (SP) induces thermal hyperalgesia, whereas hemokinin-1 (HK-1), a member of the same tachykinin family as SP, hardly induces thermal hyperalgesia; however, the underlying mechanism remains to be elucidated. Therefore, we aimed to clarify which amino acid of these peptides contributes to the induction of thermal hyperalgesia. When two chimera peptides between the N-terminal region of SP and the C-terminal region of HK-1, and vice versa, SP (1-5)/HK-1 and HK-1 (1-5)/SP, were intrathecally administered, SP (1-5)/HK-1 induced thermal hyperalgesia whereas HK-1 (1-5)/SP had hardly any effect; furthermore, thermal hyperalgesia was induced by only C-terminal fragments of HK-1 and SP. These findings indicate that the N-terminal region of HK-1 is involved in the non-induction of thermal hyperalgesia. Next, we synthesized and intrathecally administered these chimera peptides in which part of the N-terminal region of HK-1 was replaced with that of SP, and vice versa, and all synthesized peptides induced thermal hyperalgesia. Both SP (1-2)/HK-1 and HK-1 (1-4)/SP certainly induced thermal hyperalgesia, although HK-1 and HK-1 (1-5)/SP had hardly any effect; therefore, it is probable that Ser at the 2nd position and Arg at the 5th position of HK-1 may be involved in the non-induction of thermal hyperalgesia. Furthermore, peptides in which amino acid at the 3rd and/or 4th positions of HK-1 was replaced with that of SP were synthesized. Intrathecal administration of HK-1 (1-2,4-5)/SP, but not HK-1 (1-2,5)/SP and HK-1 (1-3,5)/SP, hardly induced thermal hyperalgesia. These findings indicate that three amino acids, Ser, Thr and Arg at the 2nd, 4th and 5th positions of HK-1, respectively, regulate the induction of thermal hyperalgesia by HK-1.

Central endogenous vasopressin induced by central salt-loading participates in body fluid homeostasis through modulatory effects on neurones of the paraventricular nucleus in conscious rats.

Peripherally secreted arginine vasopressin (AVP) plays a role in controlling body fluid homeostasis, and central endogenous AVP acts as a neurotransmitter or neuromodulator. The limbic system, which appears to exert an inhibitory effect on the endocrine hypothalamus, is also innervated by fibres that contain AVP. We examined whether central endogenous AVP is also involved in the control of body fluid homeostasis. To explore this possibility, we examined neuronal activity in the paraventricular nucleus of the hypothalamus (PVN), periventricular parts of the PVN and limbic brain areas, as well as AVP mRNA expression in the PVN and the peripheral secretion of AVP after central salt-loading in rats that had been pretreated i.c.v. with the AVP V(1) receptor antagonist OPC-21268. Neuronal activity in the PVN evaluated in terms of Fos-like immunoreactivity (FLI), especially in the parvocellular subdivisions, was suppressed. On the other hand, FLI was enhanced in the lateral septum, the bed nucleus of the stria terminalis and the anterior hypothalamic area. Similarly, AVP mRNA expression was enhanced in the magnocellular subnucleus of the PVN, despite the lack of a significant difference in the peripheral AVP level between OPC-21268- and vehicle-pretreated groups. We recorded renal sympathetic nerve activity (RSNA) as sympathetic nerve outflow during central salt-loading. The suppression of RSNA was significantly attenuated by i.c.v. pretreatment with OPC-21268. These results suggest that the suppression of RSNA during central salt-loading might be the result of a decrease in neuronal activity in the parvocellular subdivisions of the PVN via the inhibitory action of central endogenous AVP. The parvocellular and magnocellular neurones in the PVN might show different responses to central salt-loading to maintain body fluid homeostasis as a result of the modulatory role of central endogenous AVP.

Pharmacological characterization of desensitization in scratching behavior induced by intrathecal administration of hemokinin-1 in the rat.

Desensitization is induced by the repeated administration of high doses of substance P (SP) or hemokinin-1 (HK-1). However, little information is available about the mechanisms involved in the induction of desensitization by these peptides. Thus, to characterize this desensitization, we examined the dose-dependent effect of these peptides, the effect of pretreatment with neurokinin 1(NK1) receptor antagonists, and the effect of pretreatment with inhibitors of protein kinases such as protein kinase A (PKA), protein kinase C (PKC), calcium/calmodulin kinase II (CaMKII) and mitogen-activated protein kinase kinase (MEK). The number of scratchings induced by 10(-3)M SP or HK-1 decreased following pretreatment with 10(-11)-10(-3)M SP or HK-1 with a marked reduction at 10(-3) and 10(-6)M SP or HK-1. The effect of NK1 receptor antagonists on desensitization induced by pretreatment with 10(-6)M SP was marked, whereas there was little effect of pretreatment with these antagonists on 10(-6)M HK-1-induced desensitization. Additionally, 10(-6)M SP- and HK-1-induced desensitization was attenuated by pretreatment with PKA, PKC and MEK inhibitors, except a CaMKII inhibitor that inhibited SP-induced desensitization. These results indicate that the receptor and kinases involved in HK-1-induced desensitization are partially different from those of SP.

H-reflex and reciprocal Ia inhibition after fatiguing isometric voluntary contraction in soleus muscle.

This study investigated the H-reflex and reciprocal Ia inhibition during fatigue in the human soleus muscle. Ten healthy subjects participated in this study, and performed intermittent isometric voluntary contraction of the ankle plantarflexion at 50% MVC as the fatiguing task. Reciprocal Ia inhibition was evaluated by the degree of H-reflex amplitude depression in the soleus muscle by the test stimulus following conditioning stimulus to the common peroneal nerve. The difference in H-reflex amplitude between before and after fatiguing task was also checked. There was no significant difference in the degree of H-reflex amplitude depression, although the H-reflex amplitude significantly decreased after the fatiguing task (p < 0.01). From the results of this study, it was considered that the decrease in H-reflex amplitude was caused by descending inhibitory input from the supraspinal to alpha-motoneuron, and the excitability of the Ia inhibitory interneuron was not involved. It was suggested that the function of reciprocal Ia inhibition was difficult to modulate during fatigue caused by isometric voluntary contraction in this study.

M wave and H-reflex of soleus muscle before and after electrical muscle stimulation in healthy subjects.

PURPOSE: This study was the preliminary research for applying the evaluation of muscle fatigue using the evoked electromyography in the field of physical therapy. In this study, we speculated that muscle fatigue was induced by electrical muscle stimulation (EMS). And we studied the M wave and amplitude ratio of H/M before and after EMS because the M wave and amplitude ratio of H/M have been often used as the parameter in the study of muscle fatigue. METHODS: Subjects were five healthy males. In this study, the intermittent EMS (30 Hz) was administered to the soleus muscle of dominant leg for 10 minutes and we analyzed the amplitude of maximal M wave and the amplitude ratio of H/M in this study. RESULTS: The amplitude of maximal M wave after EMS significantly decreased compared with that before the EMS (p < 0.05). And the results of amplitude ratio of H/M varied as follows; decrease in two subjects, increase in one subject and unchanged results in two subjects. DISCUSSION: M wave reflects the excitability of muscle membrane related to the change in force during muscle fatigue and the amplitude ratio of H/M has been considered as the index of a relative excitability of alpha motoneuron pool. From the results of this study, we considered that muscle fatigue was induced by EMS as predictability because the amplitude of maximal M wave significantly decreased after EMS. And it was considered that the excitability of spinal neural function corresponding to fatigued soleus muscle by EMS was not consist change in this study. Therefore we thought that results of amplitude ratio of H/M might be influenced by excitability of spinal neural function in subject's ordinary state. CONCLUSION: It was suggested that the muscle fatigue was induced because the amplitude of maximal M wave significantly decreased after EMS in this study. And also it was suggested the excitability of spinal neural function corresponding with fatigued soleus muscle by EMS was not consist change in this study.

Test-retest reliability for recording the Erb's point potential with a change of recording posture.

This study was the preliminary research for testing the median nerve somatosensory evoked potentials (SEPs) with a postural alteration in the field of rehabilitation medicine. The purpose of this study was to investigate test-retest reliability for recording the median nerve SEPs with a change of recording posture. Subject was one healthy male, with an age of 28 years and a height of 176 cm. Median nerve SEPs was recorded in supine position on the bed with changing the tilting angle, which was set at each of 0, 15, 30, 45, 60, 75, and 90 degrees. We compared a coefficient of variation of the latency and amplitude of the Erb's point potential at each tilting angle. A coefficient of variation of the latency and amplitude at each tilting angle was calculated ranged from 10.0% to 20.0%. From a result in a coefficient of variation, all Erb's point potentials of median nerve SEPs with a change of recording posture were recorded reproducibly in this study.

The silent period from soleus and gastrocnemius muscles in relation to conditions of standing.

To clarify the relationship between postural instability and silent period (SP), we studied the variation of SP from soleus (SOL) and gastrocnemius (GAS) muscles on various standings. Subjects were eight healthy males, with a mean age of 23.5 +/- 2.2 (21-27) years. Standings in this study was regulated with ten kinds of situations provided by visual information, supporting or not by a finger and a width of base of support. SP evoked by single stimulation to tibial nerve at the popliteal fossa was recorded from SOL and GAS of dominant side during ten kinds of standings in random. The raw data were averaged 30 times. SP was calculated the duration from the artifact due to electrical stimulation to re-starting the electromyographical bursting of tonic muscle contraction under 100 or 200 microV|div on a screen. As a result of this study, there were not any statistical significant changes in SP from both SOL and GAS (one-way ANOVA: F = 1.797, F = 1.786) among ten kinds of standing. It is thought that a variation of SP may reflect the magnitude of facilitation or disinhibition of the central nervous system including spinal, brainstem or motor cortex. As the result of this study in healthy persons, it was suggested that the degree of facilitation or disinhibition of central nervous system related to SP from SOL and GAS was not different on ten kinds of standings regulated by visual information and a width of base of support.

Predominance of G3B and G14 equine group A rotaviruses of a single VP4 serotype in Japan.

A total of 65 equine group A rotaviruses (GAR) isolated from diarrheal foals at 48 farms in Hokkaido, Japan, between 1996 (29 isolates) and 1997 (36 isolates) were characterized for their VP7 and VP4 serotypes by PCR, nucleotide sequencing, and virus neutralization (VN) tests. By PCR VP7 typing, all isolates were classified as G3 or G 14, and the predominant serotype in each year was G3 (86%) in 1996 and G14 (53%) in 1997. VN tests with these 20 isolates randomly selected confirmed the specificity of PCR on the bases of complete agreement of the results in these methods (9 G3 and 11 G14), and revealed that all 9 G3 isolates were subtype G3B. There were five differing amino acid residues in three VP7 antigenic regions between subtypes G3A and G3B. Antiserum to a baculovirus recombinant that expressed P[12] VP4 neutralized all isolates and P[12] reference strains. These results suggest that genotype P[12] GAR belong to a single VP4 serotype, and that one VP4 and two VP7 serotypes (G3B and G14) of GAR were predominant in the equine population in Japan.

[Perioperative management of the patient with von Willebrand's disease].

A 67-year-old man with von Willebrand's disease, was referred to our hospital for operation of the lung cancer. He underwent right upper lobectomy of the lung and mediastinal lymph node dissection under general anesthesia. Three days before surgery, 1-desamino-8-D-arginine-vasopressin (DDAVP) was infused with good response of bleeding time shortening from 6 minutes to 3 minutes. Therefore, immediately before operation, DDAVP was infused. During the operation bleeding tendency was observed. Heat-treated factor VIII concentration and fresh frozen plasma were administered. Bleeding tendency was controlled. Total blood loss was 613 ml. During intraoperative and postoperative period, factor VIII activity and von Willebrand factor (vWF) activity were kept at adequate levels (factor VIII: 105-150%; vWF: 65-225%). The postoperative course was uneventful and he was discharged 18 days after the operation.

Intracranial self-stimulation induces Fos expression in GABAergic neurons in the rat mesopontine tegmentum.

The cholinergic neurons which originate in the mesopontine tegmentum and innervate the midbrain ventral tegmental area have been proposed to play a key role in intracranial self-stimulation reward. This mesopontine area also contains GABA neurons. Detailed information is still lacking, however, about the relationship of cholinergic and GABAergic neurons in this region to self-stimulation reward. Therefore, using double immunostaining for Fos as a marker of neuronal activity and choline acetyltransferase as a marker of cholinergic neurons, or for Fos and GABA, we investigated whether self-stimulation of the medial forebrain bundle induces Fos expression within cholinergic and GABAergic neurons in two regions of the mesopontine tegmentum, i.e., pedunculopontine tegmental nucleus and laterodorsal tegmental nucleus. Self-stimulation of the medial forebrain bundle for 1 h induced a large increase in the number of cells expressing Fos in both the pedunculopontine tegmental nucleus and laterodorsal tegmental nucleus, when compared to control brains. However, the self-stimulation-induced expression of Fos was restricted mostly to GABA-, but not choline acetyltransferase-, immunostained cells. We also examined, using microdialysis, whether self-stimulation increases acetylcholine efflux in the ventral tegmental area, a terminal region of the mesopontine tegmentum cholinergic pathway. One hour of self-stimulation significantly increased acetylcholine efflux from this terminal area. These results indicate that intracranial self-stimulation of the medial forebrain bundle may increase acetylcholine release without affecting expression of Fos in cholinergic neurons, while the same stimulation may induce Fos expression in GABAergic neurons of the mesopontine tegmentum. GABAergic as well as cholinergic neurons in this area appear to be activated by self-stimulation reward in the medial forebrain bundle.

Remission and recurrence of hyperthyroid Graves' disease during and after methimazole treatment when assessed by IgE and interleukin 13.

We analyzed the relationship between serum IgE concentrations and the remission or recurrence of Graves' disease. One hundred seven patients with Graves' disease were treated with methimazole (MMI). Serum IgE concentration greater than 170 IU/ml was found in 41 of 107 untreated patients (38.3%). However, the presence of TSH-binding inhibiting immunoglobulin or thyroid-stimulating antibody did not correlate with the IgE concentrations. Remission was found in 20 of 41 patients with elevated IgE concentrations (48.8%) after 18 months of MMI treatment, as opposed to 53 of 66 patients with normal concentrations (80.3%) (P = 0.0014). MMI treatment was discontinued in 73 patients who were followed for 26-48 months. The recurrence of Graves' disease was found in 13 patients, whereas the remaining 60 were still in remission. The rate of long-standing remission was lower in patients with elevated than normal IgE concentration (34.1% vs. 69.7%, P = 0.0007). We also analyzed serum levels of interleukin (IL)-13. Although IL-13 was not detected in all patients, the detection rate was higher in patients without remission and in those with recurrence than in those with long-standing remission (47.1%, 38.5%, and 13.3%, respectively; P = 0.0012). More patients with elevated IgE were positive for allergic diseases and for family history of allergic diseases in their first-degree relatives. We conclude that the elevation of IgE and the higher detection rate of IL-13 are associated with both remission and recurrence of Graves' disease.

MK-801 reduces non-noxious stimulus-evoked Fos-like immunoreactivity in the spinal cord of rats with chronic constriction nerve injury.

We investigated the role of N-methyl-D-aspartate (NMDA) receptors on non-noxious stimulus-induced pain by examining the effect of MK-801, a non-competitive NMDA receptor antagonist, on Fos-like immunoreactivity (FLI) in the spinal dorsal horn by non-noxious stimulation to rats with chronic constriction injury (CCI) of the sciatic nerve. In CCI rats that did not receive the non-noxious stimulus, FLI was significantly increased in laminae V/VI of the dorsal horn at the 7th and 14th days after surgery relative to sham rats. When CCI rats received non-noxious stimuli, rubbing the plantar of the hind paw, FLI in laminae I/II at the 14th day was significantly increased relative to CCI rats that did not receive the stimulation. In sham rats, the same stimulus significantly decreased FLI in laminae III/IV and V/VI at the 7th and 14th day. When MK-801 was administered intraperitoneally prior to non-noxious stimulation in CCI rats at the 14th day after surgery, the stimulus-induced FLI in laminae I/II in CCI rats was significantly reduced. This study indicates that NMDA receptor is involved in upregulating FLI in response to non-noxious stimulation of CCI rats.

Mucosal disease induced in cattle persistently infected with bovine viral diarrhea virus by antigenically different cytopathic virus.

Four cattle persistently infected with non-cytopathic (NCP) bovine viral diarrhea (BVD) virus were challenged with cytopathic (CP) BVD virus that was antigenically different from the persistent virus. Two of the animals were injected with dexamethasone (DM) and then challenged. They developed mucosal disease on days 21 and 33 post-challenge. CP-BVD viruses were isolated from their lymph nodes but not from the sera. The isolates were antigenically different from the persistent virus and the nucleotide sequence of a 787 base region in the E2 gene was markedly different. One of the isolates was indistinguishable from the challenge virus by virus neutralization tests and the nucleotide sequence showed high homology with that of the challenge CP-BVD virus. The other two cattle, challenged with the CP-BVD virus without DM treatment, developed mucosal disease at 30 and 264 days post-inoculation. CP-BVD virus was isolated from the sera as well as the lymph nodes of the cattle and was antigenically and genetically similar to the persistent virus and different from the challenge CP-BVD virus. The present results indicate that cattle persistently infected with NCP-BVD virus can develop mucosal disease induced by antigenically different CP-BVD viruses when their cellular immunity is suppressed, although they are not immunotolerant to the virus.

Genomic and serological diversity of bovine viral diarrhea virus in Japan.

Genomic properties of 62 field isolates of bovine viral diarrhea virus (BVDV) collected from 1974 to 1999 in Japan were investigated. The 5' untranslated region (UTR) was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and the 244 to 247 base nucleotide sequences were determined. Serological properties were also characterized by the cross-neutralization test using antisera against the representative strain of the classified genotype. Using phylogenetic tree analysis, BVDV 1 was subdivided into two major clusters, BVDV-1a (29 isolates) and BVDV-1b (27 isolates). In group 1a, 3 differed from the other viruses, and were classified in a branch assigned as 1a'. However, 4 isolates (So CP/75, 190 CP, 190 NCP and KS86-1-NCP) could not be assigned to group 1a or 1b. In comparison with the published sequence data, KS86-1-NCP, 190 CP and 190 NCP were similar to the Southern Africa isolates that have recently been proposed as BVDV 1c. The 5' UTR sequence of So CP/75 was unique among those of BVDV 1, suggesting that the isolate should be classified into a new genotype. Only 2 out of 62 isolates collected in 1989 and 1990 were identified as BVDV 2. The results of the cross-neutralization test strongly supported these data, suggesting a close correlation between the 5' UTR sequence and the antigenicity of BVDV.

Immunohistochemical characterisation of Fos-positive cells in brainstem monoaminergic nuclei following intracranial self-stimulation of the medial forebrain bundle in the rat.

Fos immunostaining was used as a marker of neuronal activity following intracranial self-stimulation (ICSS) of the medial forebrain bundle (MFB) in the rat, and was combined with immunostaining for tyrosine hydroxylase (TH), serotonin (5-HT), gamma-aminobutyric acid (GABA), or NR1 (one of the glutamate N-methyl- D-aspartate receptor subunits) for purposes of neurochemical identification. ICSS induced a significant but different degree of increase in the number of Fos-immunopositive (Fos+) cells in the six brainstem monoaminergic nuclei examined, which included the ventral tegmental area (VTA), substantia nigra pars compacta (SNc), dorsal raphe nucleus (DR), median raphe nucleus (MR), locus coeruleus (LC), and A7 noradrenaline cells. Densely labelled Fos+ cells were observed in the LC following ICSS, and many of these Fos+ cells were colocalized with TH. Similarly, many of Fos+ cells in the A7 and DR/MR were colocalized with TH and 5-HT, respectively. By contrast, a smaller number of Fos+ cells was detected in the VTA and SNc following the ICSS, and in these regions the majority of Fos+ cells were not colocalized with TH. Although results among regions quantitatively differed, the ICSS induced a significant increase in the number of double-labelled cells (GABA+/Fos+ or NR1+/Fos+) in all of the VTA, DR, and LC, in which the ICSS produced an ipsilaterally weighted increase in Fos-like immunoreactivity. These results suggest that ICSS of the MFB induces differential Fos expression within monoaminergic and GABAergic neurons in brainstem monoaminergic nuclei under modulation by glutamatergic afferents.

Effects of previous noxious stimulus applied to remote areas on noxious stimulus-evoked c-Fos expression in the rat trigeminal nucleus caudalis.

Noxious stimulus-evoked c-Fos expression in the spinal dorsal horn is modulated by noxious stimuli applied previously to remote areas of the body. To confirm the existence of such modulation in c-Fos expression in the trigeminal system, changes in c-Fos expression in the trigeminal nucleus caudalis induced by formalin injection into the rat whisker pad were examined by previously injecting formalin into different areas (contralateral whisker pad, ipsilateral or contralateral forepaw) of the body. Formalin injection-evoked c-Fos expression in this nucleus was significantly reduced by previous formalin injection into the contralateral whisker pad or ipsilateral forepaw but not into the contralateral forepaw. The interval between the two injections of formalin that produced a maximal reduction of formalin injection-evoked c-Fos expression was 1 h, and the reduction of c-Fos expression was less when the interval of the two noxious stimuli was longer or shorter than 1 h. These results suggested that noxious stimulus-evoked c-Fos expression in the trigeminal nucleus caudalis is reduced by noxious stimulus applied previously to remote areas, and the reduction is dependent on the area of previous noxious stimulation and interval between the two noxious stimuli.

Experimental transmission of sheep-associated malignant catarrhal fever from sheep to Japanese deer (Cervus nippon) and cattle.

The assumption that sheep carry ovine herpesvirus-2 (OvHV-2), the causative agent of sheep-associated malignant catarrhal fever (SA-MCF), is widely accepted, albeit OvHV-2 has not been isolated. We attempted experimental contact transmission of MCF from Japanese sheep persistently infected with OvHV-2 to Japanese deer (Cervus nippon) and cattle. In Experiment 1, a deer was kept in close quarters with an infected ewe. In Experiment 2, a second deer was kept with the same ewe. In Experiment 3, two cows were each kept with two infected wethers. In Experiment 1, the deer developed clinical signs at 138 days after first contact and then died. OvHV-2 genes by polymerase chain reaction (PCR) and fluorescent antibodies to Alcelaphine herpesvirus-1 were detected in the affected deer. Moreover, sequences of PCR products (423bp), obtained by amplification of materials from the sheep and from the affected deer, coincided. These results clearly confirmed that the sheep was a carrier of OvHV-2, and that this virus had induced SA-MCF in a deer. In other experiments, no OvHV-2 infection occurred in deer and cattle during the 6-18 months periods of contact, though viral genes were detected in the nasal swabs and white blood cells of the sheep. To our knowledge, this is the first report on successful experimental transmission of MCF from OvHV-2-infected sheep to deer.