Enhancement of osteogenesis by concanavalin A in human bone marrow mesenchymal stem cell cultures.

This study investigates concanavalin A (ConA) as a novel factor that may enhance osteogenesis of mesenchymal stem cells (MSCs) in vitro. Various factors, such as cytokine bone morphogenetic protein-2 (BMP-2), have been studied for their possible promotion of MSC osteogenesis in vivo and in vitro. However, the factor that might be safer, more effective, and less expensive than these has not been determined. We therefore cultured human MSCs in osteogenic medium in the presence or absence of ConA, and used calcium assays to compare the effects of ConA and BMP-2 on MSC calcification. We also used enzyme-linked immunosorbent assay (ELISA) and quantitative polymerase chain reaction (PCR) to evaluate the expression levels of bone-specific markers. ConA and BMP-2 enhanced calcification with comparable effectiveness. The combination of ConA and BMP-2 further enhanced calcification slightly but significantly. ConA also increased osteocalcin and BMP-2 protein levels in MSC culture medium. Furthermore, ConA increased osteocalcin, RUNX2, BMP-2, BMP-4, and BMP-6 mRNA expression levels. However, the gene expression pattern of ConA-stimulated MSCs was different from that of MSCs stimulated by BMP-2. Together, these results suggest that ConA and BMP-2 enhance MSC osteogenesis via different pathways. ConA-induced bone formation in MSC cultures may be useful in regenerative medicine or tissue engineering in clinical studies, as well as in basic research on bone formation.

Older boys benefit from higher initial prednisolone therapy for nephrotic syndrome. The West Japan Cooperative Study of Kidney Disease in Children.

BACKGROUND: A long course of the initial prednisolone therapy has been shown to be more effective than standard-course therapy in reducing relapse rates in children with idiopathic nephrotic syndrome, but it is commonly accompanied by corticosteroid toxicities. There has been no study on prednisolone dosage for the effective treatment of nephrotic syndrome. METHODS: Sixty-eight children (42 boys and 26 girls) with an initial attack of nephrotic syndrome were randomly allocated into two different long-course treatment groups. Patients in Group 1 received a daily prednisolone dose of 60 mg/m2 for six weeks, followed by an alternate-day dose of 40 mg/m2 for six weeks. Patients in Group 2 had a daily dose of 40 mg/m2 instead of 60 mg/m2. RESULTS: Four children in each group did not respond within six weeks. Group 1 was associated with a significantly earlier response but more frequent corticosteroid toxicities than Group 2. Boys in Group 1 had a higher rate of sustained remission than boys in Group 2 (P = 0.0073), especially boys four years old or more (P = 0.0027), but girls did not show a significant difference (P = 0.863). Boys four years old or more in Group 1 had a course of frequent relapsing less often than those in Group 2 (2 of 13 vs. 6 of 8, P = 0.0075). CONCLUSION: These findings indicate that efficient prednisolone doses may vary between sexes and ages, and that a higher initial prednisolone therapy may be of greater benefit to older boys.

Urinary N-acetyl-beta-D-glucosaminidase excretion in term and preterm neonates.

Urinary N-acetyl-beta-D-glucosaminidase (NAG) excretion was measured in term and preterm neonates on days 1, 4, 7, 14 and 28 of life. Urinary NAG showed a peak level on day 4 or 7 in these infants. In addition, it tended to be higher with the degree of prematurity. In sick preterms who were depressed at birth and had respiratory failure, the NAG activity was further elevated during the first 2 weeks, suggesting the presence of renal tubular injury in this period. These observations thus suggest that urinary NAG may be a sensitive measure of renal maturation or damage in neonates.

Clinical course and outcome of idiopathic membranous nephropathy in Japanese children.

We retrospectively studied 12 Japanese children (8 boys, 4 girls) with idiopathic membranous nephropathy (IMN), aged 2.9-15.8 (mean 7.7) years at onset. All patients were identified through either screening or a routine urinalysis; proteinuria was present in all, haematuria, which was macroscopic in 4, in 11. Three had nephrotic syndrome (NS) at or soon after onset. Stages on electron microscopy, performed in 10 patients, were I in 3, II in 5 and III in 2. Steroids alone or with cyclophosphamide were administered to 5 patients, including the 3 patients showing NS. Complete remission of proteinuria occurred in 8 patients 0.3-1.6 (mean 0.6) years after onset, and proteinuria did not recur. After a follow-up of 1.6-11.6 (mean 5.9) years, these 8 patients were in complete remission and the remaining 4 had only mild proteinuria; none had hypertension or impaired renal function. Thus, we infer that IMN in Japanese children may have a better course and outcome than IMN in non-Japanese children. Based on a comparative study of Japanese (previously reported cases added to ours) and non-Japanese (mostly Caucasian) children with IMN, this was confirmed; it is possible that steroid therapy in Japanese patients is more effective in inducing remission of NS and preserving renal function.

Evaluation of proximal tubular function in preterm infants by urinary alpha 1-microglobulin.

alpha 1-Microglobulin is a low molecular weight protein that is relatively stable in urine of low pH. There have been few reports on urinary alpha 1-microglobulin (U-A1M) excretion in preterm infants. This study was designed to establish the ranges for U-A1M in clinically stable preterm infants and to investigate changes observed in sick preterm infants. We measured U-A1M and urinary beta 2-microglobulin (U-B2M) levels at 1, 4, 7, 14, 28 and 90 days after birth in stable preterm infants (Group 1) and sick preterm infants who were depressed at birth and required immediate resuscitation (Group 2). In Group 1 infants, both parameters were high during the first 28 days and appeared to decline thereafter. U-A1M in Group 2 infants was only significantly increased compared with Group 1 on day 1, as was U-B2M. On each day of the study, U-A1M had significant positive correlations with U-B2M for all the infants studied. The changes of the two parameters observed in Group 1 probably reflect postnatal evolution of proximal tubular function in stable preterm infants. A comparison of groups 1 and 2 shows a high prevalence of acute tubular injury at birth in sick infants and also suggests that U-A1M as well as U-B2M may be a sensitive index for detecting acute tubular damage and for following its course in preterm infants.

Clinicopathological study of patients with mesangial isolated C3d deposition in various glomerular diseases.

The clinicopathological findings of isolated mesangial C3d deposition in the absence of other complement components or immunoglobulins are summarized. 55 out of 242 individual human renal biopsies examined by immunoperoxidase microscopy had isolated C3d deposition. This group consisted of 12 patients with chronic glomerulonephritis, 8 with minimal-change nephrotic syndrome, 32 with benign recurrent hematuria, 2 with Bartter's syndrome and 1 with Raynaud's syndrome. None of these patients had a disorder of the renal function and in all the patients the disease took a benign clinical course. Light-microscopic findings indicated injuries ranging from minor glomerular abnormality to mild diffuse mesangial proliferative glomerulonephritis, and there were no other remarkable findings such as cellular crescents, global sclerosis or interstitial infiltration. By immunoperoxidase microscopy, fine granular deposits of C3d were identified only in the mesangium, and arteriolar C3 staining was seen in 31 of the 55 patients. In 38 of the 42 patients examined by electron microscopy, electron-dense deposits were identified in the mesangial matrix. These findings suggest that isolated C3d deposition is a new entity with benign features both clinically and pathologically.

Distribution of IgG subclasses in membranous nephropathy.

The distribution of human IgG subclasses among the glomerular deposits in human membranous nephropathy was examined by immunofluorescence with subclass specific monoclonal antibodies. Large amounts of granular deposits of IgG4 were identified along the capillary loops in all 12 patients, and seven patients had small amounts of IgG1 deposits. Neither IgG2 nor IgG3 deposits were detectable in any of the patients. On the contrary, all four IgG subclasses were detected in membranoproliferative glomerulonephritis and lupus nephritis with a predominance of IgG1 and IgG3. The results indicate that IgG4 is predominant in the glomerular deposits in membranous nephropathy and may play an important role in its pathogenesis.

Demonstration of C3d deposits in membranous nephropathy.

An immunoperoxidase technique for light microscopy was carried out in 16 patients with idiopathic membranous nephropathy in order to determine the role of the complement system in glomeruli. Although C3 deposits are found in 50% of the cases, C3d deposits are identified in all cases in association with IgG deposits. This suggests that C3 deposits are degraded and dissociated from immune complexes. Patients with glomerular C3 deposits showed more proteinuria than those without glomerular C3 deposits. The presence of C3 deposits indicates the importance of proteinuria in human membranous nephropathy.

Distribution volume, metabolic clearance and plasma half disappearance time of exogenous luteinizing hormone releasing hormone in normal women and women with obesity and anorexia nervosa.

Synthetic LRH was infused into normal women and women with obesity and anorexia nervosa to determine the distribution volume (DV), metabolic clearance rate (MCR) and half disappearance time (t 1/2) of plasma LRH. In normal women, the DV of LRH ws 12.1 +/- 0.9 (mean +/- SE) 1, the MCR was 1478.9 +/- 39.8 ml/min (28.5 +/- 1.2 ml/min/kg body weight) and the initial t 1/2 was 5.6 +/- 0.4 min. In obese patients the DV (20.6 +/- 1.5 l) was significantly higher than that in normal subjects (P < 0.005), but the MCR and t 1/2 were not significantly different from those in normal subjects. In patients with anorexia nervosa the DV and MCR were 6.5 +/- 1.1 l and 621.8 +/- 110.5 ml/min (17.9 +/- 2.4 ml/min/kg body weight), respectively, which were both significantly lower than those in normal subjects (P < 0.02), while the t 1/2 (7.3 +/- 0.1 min) was longer than in normal subjects (P < 0.02). These data suggest that 1) the abnormal responses of some hormones to provocation tests observed in obese patients and patients with anorexia nervosa should be evaluated in consideration of changes in the DV and metabolic clearance of hormones in these conditions, and 2) in patients with anorexia nervosa changes in MCR and t 1/2 may reflect low metabolism of LRH.

[Studies on glucagon secretion in obese rats with hypothalamic lesions (author's transl)].

Glucagon secretion was studied in rats with electrolytic lesions of the bilateral ventromedial hypothalamic area (VMH-L) under various experimental conditions. The results obtained are as follows: 1. The basal plasma level of immunoreactive glucagon (IRG) was lowered in VMH-L rats 5 and 10 weeks after the operation. Plasma IRG levels after 24-hour starvation and during the arginine load were more significantly decreased in the VMH-L rats than in the control group. 2. The basal plasma level of immunoreactive insulin (IRI) showed significant positive correlations with body weight and Lee's index in these rats. The basal plasma level of IRG showed significant negative correlations with body weight, Lee's index and basal plasma IRI level. 3. In response to the arginine load, the plasma IRI level was significantly increased in VMH-L rats immediately after the operation, and the plasma IRG level was more significantly decreased in VMH-L rats 1 week after the operation than in the control group. 4. The response of plasma IRG to the arginine load was also lowered more in VMH-L rats than in rats pair-fed for 4 weeks after the operation. 5. 15 weeks after the operation, there was no significant difference in response of plasma IRI and IRG to the subcutaneous injection of epinephrine between VMH-L and control rats. These findings indicate that hypoglucagonemia in the VMH-L rats was induced by various factors, such as disorder of the autonomic nervous system, excessive insulin release, etc. The impairment of glucagon secretion may contribute to the development of obesity observed in rats with VMH-lesions.

[Endocrine function before and after treatment in patients with anorexia nervosa (author's transl)].

In order to investigate changes in endocrine function, various tests were performed on eleven patients with anorexia nervosa. In two of them, endocrine function before and after treatment was also studied. The responses of plasma LH and FSH to LH-RH were decreased in the patients as were plasma LH-RH, and LH and FSH responses to the oral administration of clomiphene citrate. In more than half of the cases, plasma TSH showed a delayed response to TRH. The basal levels of plasma GH were elevated, and plasma GH responses to hypoglycemia or 1-arginine were lowered in half of the patients. Plasma GH showed a paradoxical rise after glucose load in three out of six cases. Plasma IRI responses to 1-arginine load were decreased in some cases, but plasma IRG responses to 1-arginine were rather excessive. Plasma IRI after glucose load showed low or delayed responses in most of the cases. After the body weight was increased to normal levels by the treatment, pituitary and pancreatic endocrine functions were normalized. These findings indicate that hypothalamic function is primarily impaired in this disease and causes nutritional disorder followed by the dysfunction of some endocrine organs. Such a possibility could be supported by the improvement of endocrine function after body weight has been recovered.