Effect of oral mizoribine pulse therapy for frequently relapsing steroid-dependent nephrotic syndrome.

AIM: To evaluate the efficacy of oral mizoribine (MZB) pulse therapy given twice a week for frequently relapsing steroid-dependent nephrotic syndrome (FR-SDNS). SUBJECTS: 16 patients with FR-SDNS with a median age of 11.6 years (range 5.1 â 17.8 years) were enrolled in the study. This study was a Phase II trial. METHODS: The dose of MZB was adjusted to achieve a peak blood level of about 3 microg/ml (10.0-19.7 mg/kg/d, maximum total dose 750 mg) in two divided doses given 2 days a week before a meal. The therapeutic benefits of MZB pulse therapy were assessed based on a comparison of the incidence of relapse (times/year) and the required daily dosage of prednisolone (PSL) before and after therapy. RESULTS: The incidence of relapse after therapy was significantly lower than that before therapy (2.4 A+/- 1.6 vs. 3.4 A+/- 1.1 times/year, p < 0.05), and the required daily dosage of PSL after therapy was lower than that before therapy (0.39 A+/- 0.26 vs. 0.47 A+/- 0.24 mg/kg/d; not significant). During the follow-up period, discontinuation of PSL was possible in 6 of 12 patients who showed a decreased rate of relapse after therapy. The age at entry into the study and the peak blood concentration of MZB of these patients were significantly higher than in four patients who did not show a decreased rate of relapse (12.3 A+/- 4.3 vs. 7.9 A+/- 2.6 years, p < 0.05; 3.00 A+/- 0.93 vs. 1.97 A+/- 0.36 microg/ml, p < 0.005, respectively). No adverse effects were observed in any patients. CONCLUSION: Our results show that MZB pulse therapy is effective in decreasing the frequency of relapse and reducing the required PSL dosage in older pediatric patients with FR-SDNS.

Non-H-2-linked control of in vivo growth of SJL/J-derived reticulum cell sarcoma in recombinant inbred strains between BALB/cKe and SJL/J mice.

This report investigated the growth of reticulum cell sarcoma (RCS) in several congenic and recombinant inbred strains between BALB/cKe female (H-2d) and SJL/J male (H-2s) mice. SJA20 mice congenic with SJL/J except at the Igh locus supported RCS growth. Recombinant mice of the H-2d haplotype did not support RCS growth. However, recombinant inbred mice of the H-2s haplotype varied in their susceptibility to permit RCS growth in vivo. These results supported the role of non-H-2 gene(s) controlling the growth of RCS. Since the recombinant strains of mice exhibited different immunologic characteristics and since RCS tumor growth depended on the ability of the mice to develop a strong antitumor proliferative response, the findings reported here suggested that non-H-2 genes control the magnitude of the syngeneic proliferative response and consequently regulate RCS growth in vivo.

Inhibition of IA positive reticulum cell sarcoma tumor cell growth in syngeneic SJL/J mice by passive administration of monoclonal anti-IA antibody.

SJL/J (H-2s) mice develop spontaneous reticulum cell sarcoma (RCS) tumors at the age of 8-11 months. The RCS tumor expresses IA antigens on the cell surface, stimulates syngeneic T-cell proliferation, and appears to depend on host cells' participation for its own growth. The present study investigates the role of passively administered monoclonal anti-IA antibody on RCS tumor growth. The administration of monoclonal anti-IAs antibody into SJL/J mice prior to tumor inoculation or at the same time as tumor transplantation resulted in a significant inhibition of tumor growth. Furthermore, pretreatment of RCS tumor with antibody prior to inoculation also resulted in tumor growth inhibition. The inhibition seen in all cases studied was tumor specific, since the use of normal ascites on antibody directed against unrelated antigens resulted in no inhibition of tumor growth. Examination of tumor cells derived from spleen and lymph nodes of antibody treated mice demonstrated that the observed inhibition of tumor growth was the result of a significant depletion of IA positive tumor cells. In contrast to other tumor systems studied to date whereby anti-IA antibody promotes tumor growth, the present findings demonstrate that passive administration of anti-IA antibodies inhibit RCS tumor growth in syngeneic mice. The possible mechanisms involved are discussed.

Regulation of reticulum cell sarcoma tumor growth in SJL/J mice by a serum inhibitor affecting T-cell proliferation.

The reticulum cell sarcoma (RCS) tumor in SJL/J mice has been shown to stimulate a strong syngeneic proliferative response. A unique characteristic of the RCS tumor is that it requires host T-cells for growth. Consequently, factors that inhibit host T-cell proliferation may have a profound effect on tumor cell growth in vivo. This report investigates the relationship between a serum inhibitor of T-cell proliferation and tumor growth. Inhibition was measured in an interleukin 2-dependent proliferation of the CTLL-2 line. Sera from mice with RCS transplantable lines or from mice with spontaneous tumors were much less inhibitory than were sera from normal syngeneic SJL/J mice or from allogeneic mice. The inhibitory activity appears to follow a circadian rhythm, because serum derived in the morning was more inhibitory than was serum derived in the evening. Serum from female mice was less inhibitory than serum from male mice. In contrast to male mice, serum from 35-week-old female mice was as inhibitory as was that from young (8- to 12-week-old) mice. The mechanism of serum inhibition in tumor-bearing mice was examined. The serum was tested for the presence of interleukin 2 which could decrease inhibitory activity, and no interleukin was found. Furthermore, absorption of the serum with CTLL-2 cells did not enhance the inhibitory effect to the level of normal mouse serum. These results suggest that tumor growth in vivo coincides with less serum inhibitor, providing an adequate T-cell response requisite for tumor growth. This corroborates the notion of RCS-host T-cell interaction necessary for tumor growth which is in part regulated by a serum inhibitor.

Detection of a transforming gene in spontaneous reticulum cell sarcoma of SJL/J mice: genetically linked and host-dependent neoplasia.

Spontaneous reticulum cell sarcoma (RCS) tumor induction occurs in 90% of SJL/J mice of 8-13 months of age. Tumor induction and growth has been shown to be under the influence of both H-2 and non-H-2 genes as well as the presence of an intact host T-cell system. We postulated that cellular oncogenes may play a role in the induction, growth, and characteristics of RCS. DNA-mediated gene transfer protocols were adopted to investigate the presence of transforming genes in DNA from RCS of SJL/J mice. High molecular weight DNA was isolated from these tumors as well as from brains and livers of control tumor-free SJL/J mice and transfected into NIH-3T3 mouse and F2408 rat fibroblast cell lines. Foci of transformed cells with a peculiar round morphology were scored in both rat and mouse cultures given tumor DNA, but not in those receiving DNA from normal tissues. DNA from first-cycle transformants was transfected in further cycles of transfection, giving rise to foci with similar morphological appearances and growth properties. These experiments suggest that a transforming gene, present in RCS spontaneous tumors, is involved in the malignant conversion of the transfected normal fibroblasts. The implication of these results with respect to the induction and growth properties of RCS is discussed.

Enhanced expression of mitochondrial genes in senescent endothelial cells and fibroblasts.

It has been suggested that some mitochondrial genes are important in cellular senescence. In order to identify the mitochondrial genes that are involved in cellular senescence, we have constructed a cDNA library from senescent human vascular endothelial cells and isolated 86 senescence-specific cDNA clones by differential screening. Among the clones, we identified four distinct mitochondrial genes including NADH dehydrogenase subunit 2 (ND2), ND3, ATPase 6 and 16S ribosomal RNA. We then compared the levels of expression of these genes in young and senescent cells by using two endothelial and two fibroblast cell strains. Northern blot and slot blot hybridization confirmed that the expression levels of ND3, ATPase 6 and 16S rRNA were elevated in senescent cells of all four strains. The expression level of ND2 was also elevated during cellular senescence in three of the four strains. Because mitochondria are actively involved in oxidative phosphorylation and respiratory functions, the altered expression levels of these genes may participate in aging processes.

Effects of vasopressin and catecholamines on the maintenance of circulatory stability in brain-dead patients.

The effectiveness and reliability of long-term control of circulatory stability in brain-dead patients by combined administration of vasopressin and catecholamine was examined in detail. Twenty-five patients were divided into three groups according to the dose of vasopressin. The first group (n = 10) received no vasopressin, the second group (n = 2) an antidiuretic dose (0.1-0.4 U/hr), and the third group (n = 13) a pressor dose (1-2 U/hr), respectively. Patients given no vasopressin or an antidiuretic dose demonstrated circulatory deterioration and cardiac arrest within a short time after brain death, despite administration of a large dose of epinephrine. All patients with a pressor dose of vasopressin, however, demonstrated stable circulation as long as vasopressin and epinephrine were administered. Five patients in whom stable circulation was maintained by this technique were randomly chosen from the third group and studied under the following four conditions: (1) neither vasopressin nor epinephrine; (2) vasopressin only; (3) epinephrine only; and (4) both vasopressin and epinephrine. Compared with the controls (neither vasopressin nor epinephrine), vasopressin only increased the total peripheral resistance index, whereas epinephrine alone increased the cardiac index. Combined administration, however, raised the mean arterial blood pressure significantly by markedly increasing the total peripheral resistance index and cardiac index. Finally, in four brain-dead patients also randomly chosen from the third group, epinephrine, norepinephrine, and dopamine were compared in their circulatory effects with a pressor dose of vasopressin. Epinephrine increased both the total peripheral resistance index and cardiac index, whereas norepinephrine increased the total peripheral resistance index, compared with the baseline (no catecholamine). The required dose of norepinephrine, however, was four times that of epinephrine. The major effect of dopamine was to increase the cardiac index. We conclude that a pressor dose of vasopressin plays a central role in circulatory stabilization of brain-dead patients, and that long-term maintenance of stable circulation for a desired length of time is possible by the combined use of vasopressin and a catecholamine. Individually, catecholamines exhibit characteristic differences. Epinephrine has significant effects on both peripheral vessels and the heart, whereas norepinephrine keeps the circulation stable by increasing the total peripheral resistance index, with a much larger dose than epinephrine. Dopamine acts primarily on the heart.

Serum concentrations of cortisone and cortisol in premature infants.

To determine the relationship between biological active cortisol and its inert metabolite cortisone accurately in premature infants, serum cortisone and cortisol concentrations were measured by reversed-phase high-performance liquid chromatography (HPLC) in a group of 232 premature infants and in a control group of 127 children and 88 adults. In the control group, serum cortisone concentrations were greater than serum cortisol levels during the first 2 months after birth; cortisol levels were higher than cortisone levels after 2 months of age. However, in premature infants, serum cortisone concentrations were greater than serum cortisol levels even after the first 2 months, and total concentrations of cortisone and cortisol were equal to those in controls. Results were then analyzed according to the equivalent gestational age of premature infants. Cortisone was predominant in premature infants older than 32 weeks of equivalent gestational age, but cortisol was higher than cortisone from equivalent gestational age 24 to 31 weeks. These findings suggest that the ability of premature infants to secrete glucocorticoids resembled that of normal controls. Also, the fetal zone of the cortex, which is associated with a predominance of cortisone, remained functional in premature infants for a longer time than in control infants. Our findings that in premature infants cortisone was predominant compared with cortisol and the sum of cortisone and cortisol was equal to that in the controls indicate that cortisone cannot be disregarded whenever the cortisol level is estimated, although cortisone itself is recognized to be biologically inactive. Simultaneous measurement of serum cortisone and cortisol concentrations is important when adrenocortical function is being determined, especially in premature infants.

A new, safe, and easy technique of atrial septal defect creation.

We created an atrial septal defect (ASD) using a Ferris-Smith-Kerrison bone punch under transesophageal echocardiographic monitoring for infants with complex congenital heart diseases, eg, transposition of the great arteries. We describe a safe and easy technique of ASD creation instead of Blalock-Hanlon operation.

An experimental study of gastric mucosal blood flow in endotoxemia of the rat, with special reference to the vagus nerve and EDRF.

Gastric mucosal lesions are an important complication in endotoxemia. In order to define the role played by the vagus nerve and endothelial-derived relaxing factor (EDRF) in gastric mucosal blood flow, an investigation was carried out on four groups of rats: a control group; a group given lipopolysaccharide (LPS, 5 mg/kg); a group given gossypol-acetic acid (gossypol), which has an injurious effect on the vascular endothelial cell; and a group given L-NG-monomethyl arginine (LNMMA). Following the administration of acetylcholine and papaverine hydrochloride (via the splenic artery) and vagus nerve stimulation in all four groups of rats, the effects of vagus nerve stimulation and EDRF on the gastric mucosal blood flow were determined with a laser Doppler rheometer. In the LPS group, the gastric mucosal blood flow was decreased after acetylcholine administration and vagus nerve stimulation. This was also the case in the gossypol group. These findings suggest that inhibition of EDRF release may be responsible for the reduced gastric mucosal blood flow observed in endotoxemia.

Urinary excretion of a large amount of bound sialic acid and of under sulfated chondroitin sulfate A by patients with the Lowe syndrome.

Urine samples from two patients with the Lowe syndrome were analyzed for sialic acid and mucopolysaccharides. The sialic acid content, relative to the creatinine content, was 4--5 times higher in these patients' urine than in normal urine. Most of the sialic acid was found in unidentified glycoproteins of high molecular weight, but the levels of sialyllactose and free sialic acid were also elevated about 2 fold. A most remarkable finding was the excretion of undersulfated chrondroitin sulfate A without other mucopolysaccharides normally occurring in urine. It is suggested that a disorder in sulfation of mucopolysaccharides is etiologically implicated in the Lowe syndrome.

Prolonged hemodynamic maintenance by the combined administration of vasopressin and epinephrine in brain death: a clinical study.

The present study attempted long term hemodynamic maintenance in 16 adult brain-dead patients, 14 with head injury and 2 with cerebrovascular accidents. In addition to respiratory and fluid management, 10 were treated with continuous infusion of epinephrine to maintain systolic blood pressure above 90 mm Hg. The remaining 6 patients each received a continuous infusion of synthetic arginine vasopressin (ADH) at a rate of 1 or 2 units/hour (285 +/- 45 microunits/kg/minute) simultaneously with epinephrine. The 10 patients treated with epinephrine alone all succumbed to cardiac arrest within 48 hours of brain death, with a mean survival time of 24.1 +/- 17.2 hours. In the patients who received simultaneous ADH infusion, a minimal dose of epinephrine of no more than 0.5 mg/hr in most instances sufficed to maintain blood pressure. Their mean survival time after brain death was remarkably prolonged to 23.1 +/- 19.1 days. In brain death, ADH plays a critical role in hemodynamic maintenance, and ADH administration permits long term hemodynamic stabilization of brain-death patients, offering increasing opportunities for organ transplantation.

Experimental atrophic rhinitis in 2 and 4 month old pigs infected sequentially with Bordetella bronchiseptica and toxigenic type D Pasteurella multocida.

Experimental infections with Bordetella bronchiseptica and/or toxigenic type D Pasteurella multocida were studied in 2- and 4-month-old primary specific-pathogen-free pigs. None of the 2-month-old pigs inoculated with B. bronchiseptica or P. multocida alone developed turbinate atrophy. All the pigs inoculated with B. bronchiseptica (10(7) CFU/head) and P. multocida (10(9) CFU/head for 5 consecutive days) together, however, developed clinical and post-mortem signs of atrophic rhinitis (AR) similar to the naturally occurring disease. Slight to severe turbinate atrophy was observed in the 4-month-old pigs inoculated with B. bronchiseptica and P. multocida (at the same concentration as above) at necropsy.

Oligonucleotide probes for Bordetella bronchiseptica based on 16S ribosomal RNA sequences.

Bordetella bronchiseptica 16S ribosomal RNA (rRNA) gene was cloned and identified. On the basis of information from computer-assisted sequence comparison of the B. bronchiseptica 16S RRNA sequences with that of other bacterial species, we constructed B. bronchiseptica-specific oligonucleotide probes complementary to variable regions in the 16S rRNA molecule. Specificity of these 32P-labeled oligo-nucleotide probes was tested in a RNA/DNA hybridization with B. bronchiseptica strains and other bacterial strains. Probe BB4 was more specific than three other oligonucleotide probes. This probe BB4 was sensitive enough to be able to detect 10(4) bacterial cells.

Exercise-induced acute renal failure with renal hypouricemia: a case report and a review of the literature.

A previously healthy 16-year-old boy developed acute renal failure following a track race at a local athletic meeting. Several hours after the run, he expressed pain in the loins with nausea and vomiting. After 3 sessions of hemodialysis, he was referred to our hospital. On admission, serum creatinine was elevated to 2.3 mg/dl without an increase in serum uric acid level. After recovery from acute renal failure (ARF), hypouricemia (0.7 mg/dl) became evident in the patient. One year later, he suffered from ARF after a track race with the highest creatinine levels of 1.1 mg/dl. In order to clarify the cause and prognosis of ARF with renal hypouricemia, we summarized the clinical features in 18 patients previously described and our patient. Serum uric acid levels after recovery from ARF were below 1.0 mg/dl in all patients. Renal biopsy in 9 patients showed acute tubular necrosis in 8 patients and uric acid nephropathy in 1. The short-term prognosis of these patients seemed good, although 5 patients needed to undergo hemodialysis in their ARF courses. However, the recurrence of ARF episodes occurred in 6 patients (31.6%) including our patient, indicating that prevention of ARF might be necessary in these patients. More information is required to establish guidance for prevention of ARF.

Segmental arterial mediolysis studied by repeated angiography.

Segmental arterial mediolysis (SAM) is a rare disease of unknown aetiology. We report the fourteenth case of SAM, but the first to demonstrate serial changes on arteriography. A 65-year-old woman with abdominal pain underwent laparotomy with resection of an abnormally beaded and narrowed segment of the right branch of the middle colic artery. Characteristic pathological findings of lysis of the arterial media with dissecting haematomas were present. Other than some post-prandial pain, the patient's post-operative course was uneventful. Serial arteriography showed various abnormalities in the trunk and branches of the superior mesenteric artery. Changes in the vessels consisted of three phases, i.e. dilatation, beading with narrowing and restoration of the smooth wall, with various modifications such as aneurysmal enlargement and occlusion.

Septicaemia and arthritis in pigs experimentally infected with Pasteurella multocida capsular serotype A.

Twenty-five caesarean-derived, colostrum-deprived (CDCD) pigs and 18 specific pathogen-free pigs, aged 8 to 14 weeks, were inoculated intranasally or intratracheally with Pasteurella multocida capsular serotype A, isolated from a severe pneumonic lesion in a growing pig. The pigs were killed for necropsy on day 6 or 14 post-inoculation (PI) or, in the case of the only fatally infected animal, examined at the time of death. One CDCD pig, inoculated intratracheally with 5 ml of a bacterial suspension containing 1.7x10(9) colony-forming-units/ml, died of septicaemia on day 1 PI. Histological lesions such as severe pleuropneumonia, thrombi in glomerular capillaries, haemorrhage of the spleen, and abscesses in the tonsillar crypts were observed. The organism was recovered from a number of sites and its antigens were detected immunohistochemically in the pneumonic lesions, blood vessels of the tissues, and tonsillar crypts in the dead pig. Pneumonia, pleural adhesions and suppurative arthritis in the extremital joints were observed grossly in 3/29, 8/29 and 7/29 intratracheally inoculated pigs, respectively. In intranasally inoculated pigs, no macroscopical abnormalities were seen; histologically, however, exudative bronchopneumonia and fibrinous pleurisy were observed in 9/14 and 4/14 pigs, respectively. No significant changes were seen in the tissues of uninfected control pigs. The organism was recovered from the lesions and P. multocida type A antigen was demonstrated immunohistochemically. The organism was rarely recovered from the liver, spleen or lymph nodes (bronchopulmonary or mesenteric). The results suggest that P. multocida capsular serotype A alone can cause not only pneumonia in pigs but also septicaemia or arthritis.

Pulmonary lymphangiomyomatosis.

Two cases of pulmonary lymphangiomyomatosis are reported and findings of high resolution computed tomography (CT) are described. CT reveals that most lesions appearing reticular or emphysematous on radiographs are actually cysts, and accurately displays the extent and distribution of cystic change of the lung. On high resolution CT, individual cystic walls are much better displayed than on routine 10 mm section CT. Further, it is possible to detect even trivial pleural effusion and mediastinal lymph node swelling by CT.

Effect of Bordetella bronchiseptica and serotype D Pasteurella multocida bacterin-toxoid on the occurrence of atrophic rhinitis after experimental infection with B. bronchiseptica and toxigenic type AP. multocida.

In efficacy tests, 7 primary specific-pathogen-free piglets vaccinated with the Bordetella bronchiseptica and type D Pasteurella multocida bacterin-toxoid were challenged with B. bronchiseptica and type A P. multocida. Severe or moderate nasal turbinate atrophy was produced in the non-vaccinated pigs, whereas, only one of the 4 pigs in the vaccinated group had slight turbinate atrophy. Other immune sera against crude toxin of P. multocida type A or D were cross neutralized. The results of the present study show that the P. multocida serotype D bacterin-toxoid is effective against atrophic rhinitis caused by toxigenic P. multocida serotype A as well as toxigenic P. multocida serotype D.

Experimental pneumonia of pigs infected with Aujeszky's disease virus and Actinobacillus pleuropneumoniae.

Experimental infections were induced out to examine whether Aujeszky's disease virus (ADV) infection in pigs results in a severe pneumonia by Actinobacillus pleuropneumoniae. Intranasal inoculation of ADV (10(6.9) median tissue culture infective dose/head) in 4-month-old primary specific-pathogen-free pigs was followed by the inoculation of A. pleuropneumoniae type 1 (10(3.1) or 10(5.1) colony-forming-units/head). The pigs inoculated with ADV alone developed clinical signs of Aujeszky's disease but not pneumonia, and those inoculated with A. pleuropneumoniae (10(3.1) CFU/head) alone did not develop clinical symptoms and lung lesions. Whereas all the pigs inoculated dually with ADV and A. pleuropneumoniae (10(3.1) CFU/head) showed severe or very severe clinical symptoms and moderate or severe pneumonia and one of them died. The pigs inoculated with A. pleuropneumoniae (10(5.1) CFU/head) alone had severe clinical symptoms and one of the 2 pigs died acutely. Furthermore, all of the 3 pigs inoculated with ADV and A. pleuropneumoniae (10(5.1) CFU/head) showed clinical symptoms and moderate or severe pneumonic lesions and one pig died of disease. It was concluded that the clinical symptoms of A. pleuropneumoniae became severer by concomitant infection with ADV.