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It has been reported that allograft rejection is mediated by a variety of adhesion molecules. Using a corneal allograft model in mice, we studied the role of very late antigen (VLA)-4 and leukocyte function-associated antigen (LFA)-1 adhesion molecules in corneal allograft rejection and the effects of monoclonal antibodies (mAbs) to them in suppressing corneal rejection. C3H/He donor corneas were transplanted into BALB/c corneal beds. The allografted mice were treated with a control mAb (M18/2), mAbs to VLA-4, or LFA-1 or their combination by i.p. injection until day 7. The expression of VLA-4, LFA-1, major histocompatibility complex (MHC) class II antigens, interleukin (IL)-2, IL-2 receptor and interferon gamma (IFNgamma) in the grafted cornea were studied immunohistochemically. Cytotoxic T lymphocyte (CTL) responses to donor alloantigens were assessed. The skins from a syngeneic donor or a third-part strain were transplanted 8 weeks after the initial keratoplasty onto the mice treated with anti-LFA-1 plus anti-VLA-4 mAbs. Fourteen of 16 allografts in non-treated mice and control mAb-treated mice became opaque by 2 weeks after transplantation. At 2 weeks, non-treated allografts showed expression of MHC class II antigens on keratocytes and mononuclear cells at the host-graft junction. Also, mononuclear cells expressing VLA-4, LFA-1, IL-2, IL-2 receptor and IFNgammawere present in the stroma at the host-graft junction. The allografts treated with either anti-VLA-4 or anti-LFA-1 alone, or anti-VLA-4 plus anti-LFA-1 remained transparent for more than 2 weeks, and the survival rates at 14 weeks was 0%, 16.7%, and 75.0%, respectively. The combined use of anti-VLA-4 and anti-LFA-1 mAbs prolonged graft survival significantly (P<0.05) at 14 weeks as compared with anti-LFA-1 mAb alone. At 3 weeks, CTL responses to donor alloantigens were depressed in mice treated with either anti-LFA-1 alone or anti-LFA-1 plus anti-VLA-4. Specific prolongation of donor-syngeneic skin was observed after treatment with the combination of these two mAbs. These results indicate that VLA-4 and LFA-1 have important roles in rejection process of corneal allografts, and that the combined use of mAbs to these molecules has remarkable effects on inducing alloantigen-specific immunosuppression in corneal transplantation.

Corneal transplantation was performed by grafting C3H/He donor corneas into BALB/c corneal beds. The allografted mice were injected either with 0.5 mg/day of anti-very late antigen (VLA)-4 antibody, anti-leukocyte function-associated antigen (LFA)-1 antibody, or 0.25 mg/day each of both antibodies on days -2, 0, 1, 3, 5, and 7. After 3 weeks, cytotoxic T lymphocyte (CTL) responses to donor alloantigens were assessed. Splenocytes in the allografted mice without treatment demonstrated greater CTL responses that those in naive mice. CTL responses were depressed in mice treated with either anti-LFA-1 alone or a combination of anti-LFA-1 and anti-VLA-4 antibodies as compared with splenocytes from allografted mice without treatment. These data suggest that CTLs play significant roles in eliciting immune response to corneal allografts.

In order to obtain a useful index that can predict the effectiveness of steroid therapy in IgA nephropathy, we investigated the relationship between steroid therapy and extracapillary change. We analysed 51 cases consisting of a group of 24 cases in which steroid was administered (initial dose: prednisolone 20-60 mg/day orally, or pulse therapy), and a group of 27 cases in which steroid was not administered. First of all, we compared these two groups in terms of a clinical improvement rate over 3, 12 and 60-month-periods after the therapy, respectively. It was found that steroid therapy led to a higher improvement rate (p < 0.01-0.05), indicating that steroid is effective for IgA nephropathy. Among the group undergoing steroid therapy, the frequency of cellular crescents (C) was significantly higher (p < 0.05-0.01) in the cases in whom steroid was effective and that of fibrous crescents or adhesion (F) was significantly lower (p < 0.05-0.01). We then assessed C/F in each case of the steroid-administered group, and studied its relationship with the improvement rate. As a result, the improvement rate was below 50% in the group of C/F < 0.25, over 50% of the group of 0.25 < or = C/F < 0.75, and 100% of the group of 0.75 < or = C/F. Accordingly, we concluded that C/F could be used a parameter for predicting the effectiveness of steroid therapy in IgA nephropathy and for determining indications for steroid therapy.

Cultured PTC clearly expressed intercellular adhesion molecule-1 (ICAM-1) under the influence of tubular basement membrane antigen (TBM)-primed lymphocytes. These TBM-primed lymphocytes also demonstrated a high cytotoxic activity against cultured PTC. A pure preparation of isolated PTC from BALB/c mouse kidney was brought into primary culture. PTC was prepared by the method of Boogaard PJ et al, and our modification. Briefly, kidney was perfused with buffer containing 0.08% (w/v) collagenase. The cortical tissue was then filtered through nylon-gauze. Viable PTC were separated from other materials by isopycnic centrifugation on a discontinuous Nycodenz gradient. The confluent monolayer of PTC showed a typical epithelial morphology with cobblestone-like cells in the center of the cell-islands. Typical dome formation was observed in PTC cultures. These cells also strongly expressed gamma-glutamyl transpeptitase activity. Coculture of PTC with syngeneic lymphocytes primed with TBM antigen induced ICAM-1 expression in PTC. The TBM-primed lymphocytes had a cytotoxic activity without complement. However, neither virgin lymphocytes nor liver antigen-primed lymphocytes had cytotoxic activity. This simple syngeneic experimental model may allow us further molecular biological examination of renal tubulointerstitial diseases.

A non-hemorrhagic metalloprotease (protease L4) was purified from the venom of Chinese Mamushi (Agkistrodon halys brevicaudus) by gel filtration and anion-exchange chromatography. Protease L4 has the molecular weight of 22,000 and its optimum pH was 8.5. The protein was stable in the pH range of 5-9 and below 40 degrees C. The proteolytic activity was inhibited by metal-chelating agents and some metal ions. Calcium ion activated the activity dose-dependently, but had only a minor effect on the thermal and pH stability. L4 showed fibrinogenase activity, hydrolyzing only the A alpha chain of fibrinogen. The protease cleaved preferentially at the N-terminal of Leu and His residues of some peptides.

To compare the visual field defects in the late stage of disease between normal-tension glaucoma (NTG) and primary open angle glaucoma (POAG), measurements taken with the 30-2 and 10-2 threshold program of the Humphrey Visual Field Analyzer were analysed by statistical methods including logistic discriminant analysis. Thirty-four eyes of 34 NTG cases and 63 eyes of 63 POAG cases whose mean deviation (MD) given by STATPAC was equal to or less than -15 dB were included and there was no significant difference in MD, refraction, or age between the two groups. The analyses demonstrated that the upper arcuate area was significantly more depressed in POAG and the inferior Bjerrum area was significantly more depressed in NTG eyes. It was also shown that the lower papillo-macular area was less affected in NTG than in POAG eyes.

Experimental autoimmune hepatitis was induced in C57BL/6 mice by immunization with syngeneic liver protein and adjuvant. Hepatitis was characterized by marked cellular infiltrates, but hepatic necrosis was mild to moderate. A small dose of endotoxin (25 micrograms/mouse) produced lethal hepatitis with elevation of serum transaminase levels in these mice. The endotoxin-induced reactions were completely inhibited by i.p. administration of FUT-175 (5 mg/kg), a synthetic protease inhibitor, 1 h before the endotoxin injection. In vitro experiments showed that two-thirds of the inflammatory infiltrates were monocyte/macrophages. Cytotoxicity against syngeneic hepatocytes was significantly increased by the addition of endotoxin (25 micrograms/ml), but the same dose of endotoxin alone had no effect on the viability of hepatocytes. The endotoxin-induced increase in cytotoxicity was prominent in the glass-dish adherent (monocyte/macrophage enriched) fraction and was also demonstrated after depletion of T-cells. However, elevated cytotoxicity did not occur when FUT-175 (> 1 x 10(-7) M) was present throughout the assay period. These results seem to indicate that the hepatotoxic effects of endotoxin are mediated, at least in part, by monocytes or macrophages infiltrating the liver following immunization of liver proteins. Our results also suggest that FUT-175 has protective effects against endotoxin-induced hepatotoxic reactions.

In order to identify the endogenous protease associated with stratum corneum (SC) desquamation, we examined properties of proteases in the stratum corneum of normal human skin. SC were obtained by tape stripping, washed in toluene and then dried. The proteolytic activity in SC was measured using peptidyl 4-methyl-coumaryl-7-amides (MCAs). The SC was dispersed uniformly in the reaction mixture with dimethylformamide and Triton X-100 and incubated with the peptidyl MCAs. The protease in the SC hydrolysed both Boc-Phe-Ser-Arg-MCA and Boc-Gln-Ala-Arg-MCA (substrates for trypsin) very effectively. The hydrolytic activity was inhibited by the serine protease inhibitors diisopropyl fluorophosphate (DFP), aprotinin, antipain and leupeptin, but not by chymostatin, a chymotrypsin inhibitor. These results show that one or more trypsin-like serine protease is present in the SC of normal human skin. Casein-acrylamide electrophoresis showed that the molecular weight of this serine protease was about 30 kDa. We have previously shown that cells dissociate from human SC sheets in a detergent mixture (N,N-dimethyldodecylamine oxide and sodium lauryl sulphate). This cell dissociation was inhibited by aprotinin and leupeptin. In addition, the proteolytic activity in the outer SC was higher than that in the inner SC, and the activity in the SC of scaly skin induced by SLS treatment was higher than that of untreated skin. These results strongly suggest that the trypsin-like serine protease described here is involved in SC desquamation.

We measured six stratum corneum sphingolipid species (ceramides 1-6) in 26 males and 27 females, and found a significant change in their percentage composition only among female subjects of different age groups. There was a significant increase in ceramide 1 and 2 with a corresponding decrease in ceramide 3 and 6 from prepubertal age to adulthood. Thereafter the ratio of ceramide 2 to total sphingolipids decreased with age in contrast to ceramide 3 which showed an increase. Such a pattern of change in the aging population is different from that observed in scaly skin experimentally induced by tape stripping. The present results suggest a significant influence of female hormones on the composition of stratum corneum sphingolipids. Moreover, the different patterns of change in sphingolipid composition of stratum corneum lipids between scales from inflammatory skin and those from aged skin also suggest that epidermal biosynthesis of sphingolipids is influenced by epidermal proliferative activity.

Sweet's syndrome (acute febrile neutrophilic dermatosis) is an unusual condition characterized by fever, polymorphonuclear neutrophil leukocytosis of the blood, thick painful plaques on the face, neck and limbs, and a dense dermal infiltrate of mature neutrophils seen histologically. Recently, this disease has also been reported in association with various malignant neoplasms and chronic inflammatory disorders. In the literature, seven cases of Sweet's syndrome associated with collagen diseases have been reported, but no cases with mixed connective tissue disease (MCTD). The first case of Sweet's syndrome associated with MCTD was herein described and discussed. A 49-year-old man was admitted to our hospital with the complaints of high fever and painful erythema on his face, neck and limbs. Six months ago, MCTD was suspected, with the presence of limited cutaneous sclerosis of the hands, Raynaud's phenomenon, polyarthralgia, an elevation of CPK value and a positive anti-RNP antibody. Just before hospitalization, he suffered a prodromal infection of the upper respiratory tract for two weeks. He was diagnosed as Sweet's syndrome by the clinical and histological features. He began receiving corticosteroid therapy (prednisolone 60 mg/day), and within a week he showed dramatic improvement in the above symptoms.

Transfer of tubular basement membrane (TBM)-primed thymocytes from BALB/c mice that had been immunized with allogeneic TBM antigen without adjuvant prevented the development of interstitial nephritis (IN) in recipient BALB/c mice that had been immunized with TBM antigen with complete Freund's adjuvant (CFA) to produce IN. TBM antigen was prepared from TBM of normal ddY mice (ddY TBM antigen). BALB/c mice were highly susceptible to IN and showed a high immune response to TBM antigen when they were immunized with TBM antigen in CFA. Development of IN in high responder BALB/c mice was clearly suppressed by transfer with ddY TBM-thymocytes. The anti-TBM antibody response and the proliferative response of splenic T cells to TBM antigen were also depressed by the transfer. The cell extract of ddY TBM-thymocytes had also suppressive activity on the development of IN and on the immune response to TBM antigen. This simple system without any adjuvant for inducing the thymocytes, which has strong suppressive activity on the development of IN and on the immune response to TBM antigen, may allow us to analyse the role of suppressor T cells in negative regulation of IN.

Duplex Doppler sonography (ultrasonic equipment: Toshiba SSA-270A) were performed in the patients with various renal disease (male 44, female 32) admitted to our hospital between June 1990 and August 1991. Interlobar arterial blood flow velocity was measured at the side of renal pyramid through a longitudinal scan. Both the maximum blood flow velocity (Vmax) and the minimum blood flow velocity (Vmin) were measured quantitatively and resistive index (RI: defined as (Vmax-Vmin)/Vmax) were calculated. Vmax and Vmin correlated well with the creatinine clearance (Ccr) (r = 0.56, r = 0.66 respectively), whereas RI, by which we can detect Doppler waveform changes, correlated weak with Ccr (r = 0.39). We found twenty-nine patients (40%) with an elevated RI (greater than or equal to 0.70). They had severe renal dysfunction and active pathologic findings in the tubulo-interstitial or vascular compartment of the kidney. Doppler examination of renal blood flow velocity was valuable not only to estimate renal function but also to assess pathophysiology of renal disease.

Stratum corneum sphingolipids are of particular importance in maintaining the water permeability barrier of mammalian epidermis. Free amino acids also play an important role in water retention in the stratum corneum. To clarify the way in which these substances affect scaly skin, stratum corneum sphingolipids and free amino acids collected from artificially-induced scaly skin were analysed. Scaly skin was induced by tape stripping. The total amount of sphingolipids was quantified by gas chromatography and five of sphingolipid fractions were isolated and quantified by thin-layer chromatography. Free amino acids were analysed using a high-speed amino analyser. The total amount of sphingolipid in scaly skin did not differ statistically from that in control skin. However, a significant change in the distribution of the five sphingolipid species was observed in scaly skin and the total amount of amino acids was decreased in scaly skin. These results suggest that the distribution of these five types of sphingolipid and the total amount of amino acids are responsible for scaly skin.

A case of homicidal poisoning by aconite is reported from the viewpoint of clinical forensic medicine and analytical chemistry. Jesaconitine was detected in the vomitus, stomach contents, plasma and urine at concentrations of 32.2, 5.48, 0.433 and 1.07 micrograms/ml, respectively. The total amount of jesaconitine in the stomach contents was 1.3 mg. Macroscopic autopsy revealed hemorrhagic pulmonary edema, and histologically, diffuse contraction-band necrosis was evident in the myocardium. Exogenous administration of adrenaline or endogenous release of catecholamines are known to induce hemorrhagic pulmonary edema and contraction-band necrosis, and aconitine is reported to cause release of endogenous catecholamines centrally and to induce neurogenic pulmonary edema and ventricular-type arrhythmia. Therefore, the chief cause of death in this case was considered to be aconite-induced centrogenic arrhythmia.

We have estimated the gas concentration in an upstairs room following leakage of gas from a ground-floor room. The method used to calculate the gas concentration in the upstairs room was based on a ventilation model for two rooms. The assumptions on which the calculation was based were: 1) Uniform diffusion of gas. 2) Presence of slight openings in the room wall. 3) Reported data on the volume of the openings between the downstairs ceiling and the upstairs floor. 4) Calculation of the ventilation rate based on buoyant force. The upstairs gas concentration became steady within 4-7 h after the leakage. The ventilation rates for the 1st and 2nd floors at a steady state were 0.86 and 0.63 (room volume/h), respectively. The volume of the room did not affect the gas concentration at the steady state.

It seems that active orthodontic therapy using multi-bracket appliances presents effective assurance of a physiological stable occlusion for TMJ dysfunction syndrome in adolescents. The efficiency and problems concerning the progress of active orthodontic therapy after splint therapy were examined from the data of the change of occlusal conditions, and the function of masticatory muscles before and after therapy for TMJ dysfunction from four cases for which we performed orthodontic therapy activity. 1. All of the occurring symptoms of the TMJ dysfunction syndrome (limitation, pain, sound) in the progress of orthodontic therapy were concerned with using intermaxillary elastic. The symptom which occurred in process of using only intra-maxillary elastic was just a malaise of the TMJ. Also the sound of the TMJ relapsed in the process of using class III intermaxillary elastic and recurred after the multi-bracket appliance was removed. In this case, the condylar vertical position after orthodontic therapy changed from the lower position to the center position compared with the first visit. 2. Malocclusion was improved with orthodontic therapy in all four patients. 3. The function of the masticatory muscles after orthodontic therapy was improved in comparison with the first visit by evaluation of the masticatory muscles activity and the state of silent period.

We made a follow-up survey on the TMJ dysfunction syndrome patients who visited our clinic, using questionnaires for the purpose of raising the dental management of TMJ dysfunction syndrome patients and obtained as following results. 1. The percentage of questionnaires which were sent back was 61.5% with the finished group, and was 38.9% with the discontinued group. The average percentage for all subjects was 50.7%. 2. From the results from all subjects who answered the questionnaire and sent them back, it was found that the ratio of the patients who had understanding of the explanation before the treatment of the TMJ dysfunction syndrome was 60.5%. The ratio of the patients who answered "I could not use the splint completely" was 2.6%, and the ratio of the patients who answered "I used the splint completely in spite of malaise when using it" was 84.2%. The ratio of the patients who answered "I felt that treat time was long" was 57.9%. The ratio of the patients who had a relapse of the symptom after treatment was 33.3%, but 22.2% of their symptoms were temporary and soon disappeared. 3. There were 4 questions which showed statistical significant differences between the finished group and the discontinued group. They were the attitude to the treatment after examination at first visit, period of time the splint was used, complaint of discomfort while using the splint, concerned mind of visiting interval. 4. The rate of the patients who complained of the limitation of jaw at the first time was higher with the finished group than with the discontinued group.

The ambulatory ECG system has been used not only in detecting arrhythmia but also in the diagnosis of myocardial ischemia. Therefore, it is necessary to reproduce ECG waveforms as accurately as possible. However, ambulatory ECG waveforms which are in clinical use are apt to be distorted because the frequency responses do not satisfy the AHA (American Heart Association) recommendation which is well accepted as the standard of the conventional ECG system. An improvement of frequency response is reported here using a compensation circuit, whose theory is based on the natural observation method. Since our system is adjusted with square waveforms and composed of a simple analog circuit, it can be compensated easily in real time. In this paper, our newly developed system and various improved ECG signals are presented and compared with conventional ECG signals. As a result, we could obtain less distorted waveforms than from commercially available devices. This new approach enables us to obtain diagnostic information of the distortion-free ST segment in ECG waveforms.

The clinical records and treatment results of 163 patients with Hodgkin's disease, who were seen at Ellis Fischel State Cancer Hospital (EFSCH) between 1940 and 1971, were reviewed and analyzed. More than 200 clinical and histological variables were recorded for each case of Hodgkin's disease, including details of radiotherapy and chemotherapy. Statistical studies were carried out in order to evaluate the independent prognosis significance of each of these factors. All of the lesions were reclassified according to the Lukes proposal which was modified and recommended at the 1965 Rye classification (except for hepatomegaly which was included in Stage IV). This is a retrospective study, and the modern techniques of staging were rarely used in pretreatment studies (since 1965, only ten patients have had an abdominal exploration). The basic work-up consisted of a complete blood count, urinalysis, blood type, chest X ray, and EKG. Lymphangiogram and radioisotope liver scans were used on less than 10% of the patients. About 30% of the patients had gastrointestinal X rays and 70% had IVP. Bone marrow biopsies -- the majority of which were done by needle aspiration -- were obtained for approximatley 50% of the patients. Clinical stage, histological type, and presence of absence of systemic symptoms appeared to be themost significant prognostic factors. The classification of systemic symptoms according to the criteria of either the Rye or Ann Arbor conferences showed no particular difference in determining the survival rate. Among the systemic symptoms, fever appeared to be the most important for survival rate. Survival rates were higher in nonanemic and nonlymphocytopenic patients. Eosinophilia, blood group, and Rh factor had no prognostic significance. The relapse-free interval was an important indicator of long-term prognosis. The unfavorable influence of relapse in ultimate prognosis was clearly seen; however, the extent of the relapse site was shown to have no significant influence on survival.