Correlations between interleukin-8, and myeloperoxidase or luminol-dependent chemiluminescence in inflamed mucosa of ulcerative colitis.

Interleukin-8 (IL-8) is a peptide which induces not only chemotaxis of neutrophils but also the release of reactive oxygen metabolites from the neutrophils. There are few reports which clarify the relationships between IL-8 and mucosal infiltration of neutrophils or reactive oxygen metabolites produced by neutrophils in the colonic mucosa of ulcerative colitis (UC). Biopsy specimens of colonic mucosa obtained from 26 patients with active UC and 21 patients with inactive UC were studied in order to clarify the relationships among the inflammation factors in UC. Levels of IL-8 and myeloperoxidase in organ culture media of the biopsy specimens from active UC (measured by ELISA and EIA) were significantly higher than those from inactive UC and controls. Reactive oxygen metabolites of biopsy specimens in active UC (measured by luminol-dependent chemiluminescence) were also markedly increased compared to those in inactive UC and controls. The levels of IL-8 were closely correlated to luminol-dependent chemiluminescence or myeloperoxidase levels. However, the levels of IL-8 and myeloperoxidase did not correlate with the grades of activity on colonoendoscopic findings. These findings suggest that IL-8 may play a role in the pathophysiology of UC but it does not define the endoscopic activity grades of UC.

Spectrum of cytokine gene expression in intestinal mucosal lesions of Crohn's disease and ulcerative colitis.

BACKGROUND/AIMS: We investigated the mRNA expression of spectrum of cytokines in the colonic mucosa in inflammatory bowel disease (IBD). METHODS: The expression of cytokine gene was evaluated by using the reverse transcription-polymerase chain reaction and the radioactivity of amplified cDNA standardized by coamplified beta-actin cDNA. RESULTS: Crohn's disease and ulcerative colitis showed significantly increased expression of IL-1beta, IL-6, IL-8 and TNF-alpha mRNA as compared with controls (p < 0.05). CONCLUSION: Proinflammatory cytokines such as IL-1beta, IL-6, IL-8 and TNF-alpha are closely involved in the immune abnormalities of inflammatory mucosal lesions in IBD.

Study of cytokines in ulcerative colitis.

We investigated the lymphocyte-activation antigens and the expression of cytokine genes in the mucosa of ulcerative colitis (UC). Fresh colonic mucosal biopsy specimens from patients with UC and controls were fixed for the immunohistochemical study of CD4, HLA-DR, and CD25, and other specimens were prepared for the RNA analysis of cytokines. Gene expression was evaluated by the reverse transcription-polymerase chain reaction, and the radioactivity of dot-blotted amplified cDNA was standardized by co-amplified beta-actin cDNA. The inflamed mucosa of active UC showed increased CD4+DR+ and CD25+ cells in comparison with control subjects. Active UC showed significantly increased mRNA expression of IL-1 beta, IL-2R alpha, IL-6, IL-8, and TNF alpha compared with the controls. We found no significant difference in the mRNA expression for IL-2, IL-4, IL-10, and IFN-gamma between active UC and controls. Increased CD4+DR+ and CD25+ cells in active UC mucosa indicate mucosal CD4(+) T cell activation in the lamina propria, but we did not clarify Th1 or Th2 specific T cell activation from our study of cytokine mRNA expression. The increased mRNA expression for IL-1 beta, IL-6, and TNF alpha in the mucosal lesions of UC indicates that these inflammatory cytokines may play important roles in the pathogenesis of UC.

Research on oxygen transport in jaw cysts: investigation of changes of erythrocyte membrane fluidity.

I have investigated oxygen transport in the fluid from jaw cysts. Measurements were made of erythrocyte membrane fluidity, mean corpuscular volume, PO2, leucocyte percentage and erythrocyte and leucocyte numbers, both before and after irrigation of the cysts with antibiotics. On the basis of the results it is suggested that the membrane fluidity of erythrocytes in jaw cysts is decreased by infection and the PO2 value falls as a result of this decrease and also because of the increase in mean corpuscular volume and the raised oxygen consumption due to neutrophilia within jaw cysts.

[Effects of d-pseudoephedrine on tracheo-bronchial muscle and the cardiovascular system (author's transl)].

The action of d-psuedoephedrine on bronchial smooth muscle, respiratory resistance and blood pressure was compared with that of 1-ephedrine. The following results were obtained. 1) Administration of dl-isoproterenol on isolated guinea pig tracheal muscle previously constricted with acetylcholine (ACh) or histamine (his) was the most effective and d-pseudoephedrine had the same effect as 1-ephedrine. The relaxing effect of dl-isoproterenol, 1-ephedrine or d-pseudoephedrine on ACh or His induced tracheal constriction was competitively antagonized by the pretreatment of propranolol. 2) The intravenous or sublingual administration of d-pseudoephedrine on the increased respiratory resistance induced by His produced the same effect as 1-ephedrine. 3) The increase in blood pressure with d-pseudoephedrine was weaker than that of 1-ephedrine. 4) When 1-ephedrine of d-pseudoephedrine was repeatedly injected into the same animal, signs of tachyphylaxis were observed. It was found that d-pseudoephedrine, the main alkaloid of Ephedrine Helba, exhibited the relaxing effect on bronchial smooth muscles as did 1-ephedrine. The respiratory resistance increased by His was inhibited by the sublingual administration of d-pseudoephedrine and 1-ephedrine.

[Effects of diazepam on rage reaction elicited by the lateral hypothalamus (LH) and on single unit activities in the LH and the basal medial amygdaloid nucleus (Abm) in cats (author's transl)].

Effects of diazepam were examined on the whine reaction elicited by LH stimulation and on unit activities in the LH and Abm in cats. The spontaneous firing frequency of Abm neurons was 5 to 30 spikes/sec and in all 6 neurons isolated the firing frequency increased by non-nociceptive and/or clap-stimulation. Diazepam decreased the spontaneous firing frequency of all Abm neurons isolated and the increased firing frequency elicited by non-nociceptive and/or clap-stimulation was also depressed by diazepam. The spontaneous firing frequency of neurons in the LH was 4 to 5 spikes/sec and all 6 neurons isolated firing frequency increased by non-nociceptive stimulation. Only one of 6 neurons, however, was activated by clap-stimulation. Diazepam decreased the spontaneous firing frequency of all LH neurons. Out of 6 neurons responsive to non-nociceptive stimulation, 3 were also depressed by diazepam. The other neurons were not affected by diazepam. These results suggest that depressed action of diazepam on the whine reaction elicited by the LH stimulation may be related to the decrease of firing in the Abm and/or the LH by diazepam.

[Effects of diazepam on evoked potential recorded from basal medial amygdaloid nucleus, lateral hypothalamus and midbrain reticular formation (author's transl)].

The evoked potential in the lateral hypothalamus (LH) recorded by stimulation of basal medial amygdaloid nucleus (Abm) showed a triphasic pattern and diazepam (2 mg/kg, i. p.) decreased the late component. The evoked potential in the midbrain reticular formation (MRF) recorded by stimulation of Abm showed a fast component with a relatively short latency followed by a biphasic late component and diazepam decreased the late component. Though the evoked potential in the Abm recorded by stimulation of LH showed a triphasic pattern, diazepam had no influence on the amplitude. Diazepam increased markedly the amplitude of evoked potential in the MRF recorded by stimulation of LH. Diazepam was ineffective on the evoked potential in the Abm recorded by stimulation of MRF. Diazepam decreased markedly the late component of evoked potential in the LH recorded by stimulation of MRF. These results suggest that the depression of emotional behavior by diazepam may be particularly related to the fact that the evoked potential in the LH recorded by stimulation of Abm was decreased by diazepam.

[Effects of morphine on the single unit activity of nucleus dorsal raphe].

Effects of morphine were studied in 36 gallamine triethiodide immobilized adult cats under light N2O anesthesia. Single units were recorded from the nucleus dorsal raphe using a stainless steel microelectrode. Dorsal raphe neurons were divided into two types; one was a clock-like (CL) neuron which was typically slow in rate, rhythmic and stable throughout recording time, and the other was a non-clock like (NCL) neuron which was relatively irregular in pattern as compared with CL neuron. Out of 36 neurons recorded in this experiment, 13 were CL neurons and 23 were NCL neurons. Mean discharge rates (spikes per sec) of CL neuron and NCL neuron were 2.84 (range of 2.01 approximately 3.68) and 4.11 (range of 0.10 approximately 35.09), respectively. None of the 13 CL neurons responded to the nociceptive (pinch and/or brdaykinin) and non-nociceptive (hair bending and/or tapping) stimuli. On the contrary, out of 23 NCL neurons, 13 responded to both nociceptive and non-nociceptive stimuli, 4 were responsive to only non-nociceptive stimuli and 6 were not responsive to these stimuli. Receptive fields of NCL neurons were wide with various somatic modalities. The latency for bradykinin (3 microgram) was 6.46 +/- 0.77 sec. Effects of morphine were examined in 6 CL neurons and 12 NCL neurons. After morphine, no nociceptive neuron responded to nociceptive stimuli, although there was no appreciable change in the responsiveness to non-nociceptive stimuli. Firing frequency and pattern of all CL neurons were unaffected by morphine. Out of 12 NCL neurons, 7 were unaffected in firing frequency and 5 were decreased after morphine.

[Effects of a benzodiazepine derivative, MS4101, on emotional behaviour of untamed cats].

Effects of MS4101 on emotional behaviour in untamed cats were studied and compared with those of diazepam. Offensive behaviour, i.e., whine response to a rod presented in front of the snout and blowing air on back hair was markedly observed, and whine, attacking and biting responses to tapping with a rod on the back in these cats were marked. Defensive behaviour, i.e., hissing, crouching body, ear flattening to blowing air on back hair, a rod presented and tapping was markedly observed. From 30 min after MS4101 and diazepam in doses of 2 approximately 4 mg/kg i.p., offensive behaviour in untamed cats was depressed. ID50 (50% of inhibition dose) of offensive behaviour for MS4101 and diazepam was 2.40 (1.95 approximately 2.95) mg/kg i.p. and 0.96 (0.69 approximately 1.34) mg/kg i.p., respectively. MS4101 and diazepam in doses of 2 approximately 4 mg/kg i.p. decreased the offensive behaviour. ID50 of defensive behaviour for MS4101 and diazepam was 3.00 (2.46 approximately 3.66) mg/kg i.p. and 1.45 (1.14 approximately 1.84) mg/kg i.p., respectively. Both MS4101 and diazepam exhibited muscle relaxant effects. Here, diazepam was more effective than MS4101. ED50 of muscle relaxant activity for MS4101 and diazepam was 4.30 (3.03 approximately 6.11) mg/kg i.p., 7.40 (5.04 approximately 10.66) mg/kg i.p., respectively. A single administration of MS4101 and of diazepam in doses 2 mg/kg i.p. enhanced food intake.

[Effects of MS4101, a new benzodiazepine compound, on evoked potentials induced by noxious stimulation of hind legs in cats].

The effects of 10-chloro-11b-(2'-fluorophenyl)-2,3,5,6,7,11b-hexahydro-7-(2''-hydroxyethyl)benzo[6,7]-1,4-diazepino [5,4-b]oxazol-6-one (MS4101), a new benzodiazepine compound, on evoked potential in cats were examined and compared with diazepam. Though evoked potential in somatic cortex I, motor cortex I and posterior groups induced by tibial nerve stimulation were unaffected by a small dose of MS4101 (1 mg/kg) which did not influence on limbic system, the amplitude of evoked potential in midbrain reticular formation was reduced. The amplitude of evoked potential in amygdala and hippocampus induced by cutaneous nerve stimulation was reduced by MS4101 and diazepam. These two drugs had the same effect on amygdala and difference in the degree of action between these drugs was not observed. Sham rage response following head shake, circling and whine behaviour was observed by electrical stimulation of the hypothalamus. The threshold of sham rage response by raised by MS4101 and diazepam.