Mysm1 is required for interferon regulatory factor expression in maintaining HSC quiescence and thymocyte development.

Mysm1(-/-) mice have severely decreased cellularity in hematopoietic organs. We previously revealed that Mysm1 knockout impairs self-renewal and lineage reconstitution of HSCs by abolishing the recruitment of key transcriptional factors to the Gfi-1 locus, an intrinsic regulator of HSC function. The present study further defines a large LSKs in >8-week-old Mysm1(-/-) mice that exhibit increased proliferation and reduced cell lineage differentiation compared with those of WT LSKs. We found that IRF2 and IRF8, which are important for HSC homeostasis and commitment as transcription repressors, were expressed at lower levels in Mysm1(-/-) HSCs, and Mysm1 enhanced function of the IRF2 and IRF8 promoters, suggesting that Mysm1 governs the IRFs for HSC homeostasis. We further found that the lower expressions of IRF2 and IRF8 led to an enhanced transcription of p53 in Mysm1(-/-) HSCs, which was recently defined to have an important role in mediating Mysm1(-/-)-associated defects. The study also revealed that Mysm1(-/-) thymocytes exhibited lower IRF2 expression, but had higher Sca1 expression, which has a role in mediating thymocyte death. Furthermore, we found that the thymocytes from B16 melanoma-bearing mice, which display severe thymus atrophy at late tumor stages, exhibited reduced Mysm1 and IRF2 expression but enhanced Sca1 expression, suggesting that tumors may downregulate Mysm1 and IRF2 for thymic T-cell elimination.

Free fatty acids impair hepatic insulin extraction in vivo.

Hyperinsulinemia is a common finding in obesity and results from insulin hypersecretion and impaired hepatic insulin extraction. In vitro studies have shown that free fatty acids (FFAs), which are often elevated in obesity, can impair insulin binding and degradation in isolated rat hepatocytes. To investigate whether FFAs impair hepatic insulin extraction (E(H)) in vivo, either saline (SAL) or 10% Intralipid (0.03 ml x kg(-1) x min(-1)) plus heparin (0.44 U x kg(-1) x min(-1)) (IH) was infused into normal dogs to elevate FFA levels. Insulin was infused intraportally at 18 pmol x kg(-1) x min(-1) for 150 min (period A, high insulin dose), and then at 2.4 pmol x kg(-1) x min(-1) for another 150 min (period B, low insulin dose). After the low portal insulin dose, additional insulin was infused peripherally at 8.4 pmol x kg(-1) x min(-1) for 120 min (period C) to assess the clearance of insulin from the peripheral plasma. In 16 paired experiments, FFA levels were 1,085 +/- 167, 1,491 +/- 240, 1,159 +/- 221 micromol/l (IH) and 221 +/- 44, 329 +/- 72, 176 +/- 44 micromol/l (SAL) in periods A, B, and C, respectively. Peripheral insulin levels were greater with IH (P < 0.001) than with SAL in all periods (1,620 +/- 114, 126 +/- 12, 1,050 +/- 72 pmol/l for IH vs. 1,344 +/- 168, 96 +/- 4.2, 882 +/- 60 pmol/l for SAL). Glucose clearance was impaired by IH in all periods (P < 0.05), whereas glucose production was slightly increased by IH during period B. Peripheral insulin clearance (Cl) and E(H) were calculated from the insulin infusion rate and insulin concentration data in each period by taking into account the nonlinearity of insulin kinetics. Cl was lower (P < 0.01) with IH (9.6 +/- 0.6, 12.0 +/- 0.9, 10.2 +/- 0.6 ml x kg(-1) x min(-1)) than with SAL (11.2 +/- 1, 13.6 +/- 0.7, 11.9 +/- 0.9 ml x kg(-1) x min(-1)) in periods A, B, and C. E(H) was also lower (P < 0.05) with IH (25 +/- 4, 40 +/- 5, 32 +/- 5%) than with SAL (30 +/- 2.8, 47 +/- 3, 38 +/- 3%). We conclude that FFAs can impair hepatic insulin extraction in vivo at high and low insulin levels, an effect that may contribute to the peripheral hyperinsulinemia of obesity.

Complement assembly of two fragments of the streptococcal protein G B1 domain in aqueous solution.

We examined the complementation of various pairs of fragments derived from the streptococcal protein G B1 domain by NMR and CD. Most were not associated; however, one pair of fragments (1-40) and (41-56) interacted sufficiently enough to regenerated a stable 1:1 complex, Kd = 9 x 10(-6) M. A 2D-NMR analysis showed that the structure of the complex resembled that of native domain. Here we discuss the complementation from the viewpoint of the folding pathway of the protein.

Congenital heart defects in Aarskog syndrome.

We report on 10 Japanese individuals from 3 families affected with Aarskog syndrome. Pulmonary stenosis and ventricular septal defect with spontaneous closure were detected respectively, in 2 of them as an uncommon finding. A review documented 169 non-Japanese cases (2 with congenital heart defects), while of 30 Japanese individuals reported till now, 4 (including ours) had cardiac anomalies. We propose that this combination is not coincidental and that in all cases of Aarskog syndrome a cardiac evaluation is indicated.

Desquamative interstitial pneumonia in sibs.

We report on 2 sibs with desquamative interstitial pneumonia. The female died at age 1 7/12 years despite use of prednisolone and methylprednisolone, while the male, now age 3 years. is alive with oxygen support. The occurrence of desquamative interstitial pneumonia in sibs born to normal parents suggests that in some cases the disease is an autosomal-recessive trait.

Giant hepatic granuloma caused by Bartonella henselae.

We report a 10-year-old girl with a 3.0- by 3.5-cm giant hepatic granuloma caused by Bartonella henselae. Such a solitary and large granuloma associated with B. henselae infection has not been previously reported. We believe that B. henselae infection is a consideration in the differential diagnosis of a large hepatic mass.

Minimum number of water molecules required for inclusion of d-limonene in the cyclodextrin cavity.

The ratios of inclusion of d-limonene in alpha-, beta-, and gamma-cyclodextrin (CD) cavities were measured under low moisture conditions by a micro-aqueous method, and the minimum numbers of water molecules required for inclusion of d-limonene in alpha-, beta-, and gamma-CD cavities were determined by the auto-catalytic inclusion model. The minimum number of water molecules required for alpha-, beta-, and gamma-CD inclusion calculated from the proposed model were comparable with that in the CD cavity in the crystal state. The effect of ethanol upon the inclusion process was investigated, particularly with a low water content. This is the first study involving measurement of the minimum number of water molecules required for the inclusion of organic molecules (d-limonene) in the CD cavity.

Function and distribution of a steroidogenic cell-specific transcription factor, Ad4BP.

Ad4BP was identified as an essential transcription factor regulating steroidogenic cell-specific and cAMP-dependent transcription of the genes of steroidogenic P450s. The Ad4BP transcript was detected in steroidogenic tissues such as adrenal gland, testis, ovary, placenta and brain by RT-PCR, and showed good correlation with the expression of steroidogenic P450s. The genes of steroidogenic P450s, which are transcribed only in steroidogenic cells, were transcribed in non-steroidogenic cells when an Ad4BP expression vector was introduced into the cells. To study the function of Ad4BP in the differentiation of the steroidogenic tissues, immunochemical and immunohistochemical studies were performed with the tissues prepared from various developmental stages of rats. Adrenal cortex expressed Ad4BP since the tissue was detected in the dorsal wall of the fetus. Gonadal tissues expressed Ad4BP in a sex-dependent manner. High levels of Ad4BP expression were detected in fetal and prepubertal testes and in prepubertal and adult ovaries, whereas low level expressions were observed in the adult testes and in the fetal ovaries. The expression of Ad4BP in the gonads correlates well with the expression of the Müllerian inhibiting substance gene as well as the steroidogenic P450 gene for both sexes. These observations indicate that Ad4BP plays an important role in the development and differentiation of the steroidogenic tissues including sexual differentiation of the gonadal tissues through activation of the transcription of its target genes.

Protection against impairment of memory and immunoreactivity in senescence-accelerated mice by acidic fibroblast growth factor.

Subcutaneous injection of aFGF once a week into senescence-accelerated mice (SAM)P8 was begun at 3 weeks after birth and continued for 10 months. Saline was injected as a control. Learning and memory and cellular immunological functions in the aFGF group were enhanced significantly, while those of the saline group deteriorated. 1. The number of cholinergic neurons was decreased by less than 20% and choline acetyltransferase activity in individual neurons in the medical septum which send monosynaptic terminals to the hippocampus was significantly decreased in the saline group, but not so much in the aFGF group. 2. The respective densities of muscarinic and NMDA receptors and the aFGF receptor, i.e., FGFR-1 on the hippocampal neurons were also significantly higher in the aFGF group than in the saline group. 3. The long-term potentiation in the hippocampal slice preparations after a brief tetanic stimulation at the Schaffer collateral/commissural afferents was significantly facilitated in the aFGF group, but not in the saline group. 4. These data indicate the normalization caused by aFGF of the medial septohippocampal circuit, which is necessary for learning and memory. 5. The delayed type hypersensitivity reactions (DTH) in the footpad caused by challenge with trinitrophenyl or sheep red blood cells as measured at the end of the 2nd and 7th months, indicated the T cell immune response. Both types of DTHs were reduced in the 7th month as compared with the 2nd month in the saline group, but the aFGF group was protected against this reduction in accordance with age. 6. These results show that aFGF provides protection against impairment of not only learning and memory but also DTH immunoreactivity in SAMP8. They also indicate a close relationship between learning and memory and T cell immune function.

Hepatic resection for hepatocellular carcinoma.

Between July 1973 and September 1988, 119 patients with hepatocellular carcinoma underwent hepatic resection at Keio University Hospital, Tokyo. Hepatic resection was performed not only for patients with liver cirrhosis and obstructive jaundice but also for patients with advanced disease. Eighty (67.2%) of the 119 patients had liver cirrhosis and four patients had obstructive jaundice. Two or more segments of the liver were resected in 56 (47.0%) patients, 29 of whom had liver cirrhosis. Eleven patients died within 30 days after surgery, an operative mortality rate of 9.2%. Seven additional patients could not be discharged from the hospital, resulting in a hospital death rate of 5.9%. Seventeen of these 18 patients had cirrhosis. Selection of patients with sufficient reserve function of the remaining liver portion, caused a great reduction of the incidence of postoperative death. The 5-year actuarial survival rate for the 101 patients who were discharged from the hospital was 39%, and 13 patients lived longer than 5 years, the longest survival period being 13 years 10 months. Hepatocellular carcinoma is amenable to hepatic resection if patients with sufficient reserve function of the liver are selected.

Carcinoma of the bifurcation of the hepatic ducts.

Thirty-one patients with carcinoma of the bifurcation of the hepatic ducts were treated during a period of ten years. Sixteen of the 31 patients underwent resection. Left, right, and extended right lobectomy with the resection of the bilateral hepatic ducts, common hepatic and bile ducts were performed on 11, 1, and 3 patients, respectively. In a cirrhotic patient, resection of the left medial segment and bile ducts was carried out. The biliary tract was reconstructed using hepaticojejunostomy or hepatojejunostomy. Four of the 16 patients are still alive for six months, 1 1/2 years, one year and 11 months, and two years and ten months, respectively. The remaining 12 patients died of recurrence at periods ranging from six months to three years and eight months. The treatment is now focused on increasing long-term survival.

Effect of metformin on advanced glycation endproduct formation and peripheral nerve function in streptozotocin-induced diabetic rats.

The effects of metformin treatment on advanced glycation endproduct formation and peripheral nerve function in streptozotocin-induced diabetic rats were examined. Streptozotocin-induced diabetic rats were treated with low dose metformin (50-65 mg kg(-1) daily) or high dose metformin (500-650 mg kg(-1) daily) for 10 weeks. While the metformin-untreated diabetic group showed a significant increase of advanced glycation endproducts (6.1-fold in the lens, 1.6-fold in the sciatic nerve, 2.3-fold in the renal cortex, and 1.9-fold in plasma; all P < 0.01) compared with the healthy control group, both metformin-treated groups had significantly less advanced glycation endproduct deposition. The % decrease in the diabetes-induced increase in advanced glycation endproduct formation by low and high dose metformin treatment was 25% and 72% in the lens (both P < 0.01), 31% and 42% in the sciatic nerve (both P < 0.05), and 16% and 33% in the renal cortex (P < 0.05 and P < 0.01), respectively. However, the plasma advanced glycation endproduct level showed no significant difference from that in the untreated diabetic group, in spite of slight decrease in plasma glucose and glycated hemoglobin levels in the metformin-treated groups. The diabetes-induced sciatic nerve conduction velocity deficits were improved by 46% and 42% by low and high dose metformin treatment, respectively (both P < 0.01). These data suggest that metformin may have a direct antiglycative action, which in turn contributes to amelioration of peripheral nerve function. Thus, metformin treatment may be effective in the prevention of diabetic complications through not only lowering plasma glucose, but also directly inhibiting advanced glycation endproduct formation.

Dynamics and modeling on fermentative production of poly (beta-hydroxybutyric acid) from sugars via lactate by a mixed culture of Lactobacillus delbrueckii and Alcaligenes eutrophus.

The mixed culture system was considered in the present research where sugars such as glucose were converted to lactate by Lactobacillus delbrueckii and the lactate was converted to poly beta-hydroxybutyrate (PHB) by Alcaligenes eutrophus in one fermentor. For the modeling of the effect of NH3 concentration on the cell growth of A. eutrophus and PHB production rates, metabolic flux distributions were computed at two culture phases of cell growth and PHB production periods. It was found that the NADPH, generated through isocitrate dehydrogenate in TCA cycle, was predominantly utilized for the reaction from alpha-ketoglutalate to glutamate when NH3 was abundant, while it tended to be utilized for the PHB production through acetoacetyl CoA reductase as NH3 concentration decreased. This phenomenon was reflected in the development of mathematical model. In the mixed culture experiments, the two phases were observed, namely the lactate production phase due to L. delbrueckii and the lactate consumption phase due to A. eutrophus. The lactate concentration could be estimated on-line by the amount of NaOH solution and HCl solution supplied to keep the culture pH at constant level. Several mixed culture experiments were conducted to see the dynamics of the system. Finally, a mathematical model which can describe the dynamic behavior of the present mixed culture was developed and the model parameters were tuned for fitting the experimental data. The model may be used for several purposes such as control, optimization, and understanding process dynamics etc.

Impairment of insulin-induced vasodilation is associated with muscle insulin resistance in type 2 diabetes.

To clarify the association between the actions of insulin on the vascular wall and on the muscles in diabetes, we evaluated insulin-mediated vasodilation and muscle glucose uptake simultaneously using the euglycemic hyperinsulinemic glucose clamp technique and the calculation of total peripheral vascular resistance (TPR) from arterial pulse wave analysis in 19 Japanese patients with type 2 diabetes who had no signs of atherosclerosis. During the clamp study, the plasma norepinephrine (NE) level and plasma renin activity (PRA) increased without showing any significant correlation to the glucose infusion rate (GIR); a marker of muscle insulin sensitivity, and no changes of other plasma vasoactive hormone levels were observed. TPR decreased over time during the clamp study. The decrease of TPR from baseline was 0.88 +/- 0.02 at 1 h (mean +/- S.E.M., P < 0.01) and 0.79 +/- 0.03 at 2 h (P < 0.01), and the relative change in TPR from baseline was negatively correlated with GIR (r = -0.48 at 1 and 2 h; both P < 0.05). Our results suggest that there is also insulin resistance in the vascular wall, and this phenomenon may be associated with muscle insulin resistance in type 2 diabetes.

Clinical usefulness of measuring urinary polyol excretion by gas-chromatography/mass-spectrometry in type 2 diabetes to assess polyol pathway activity.

INTRODUCTION: Decreased myo-inositol levels and increased activity of the polyol pathway have been proposed to play a role in causing diabetic microvascular complications. There are few clinical methods for examining the activity of the polyol pathway in diabetic patients. We assessed the effect of changes in glycemic control on polyol pathway activity by measuring urinary polyol excretion. MATERIALS AND METHODS: Gas-chromatography/mass-spectrometry (GC/MS) was used to assess the urinary excretion of glucose and polyols (myo-inositol, sorbitol, and fructose) in 50 patients who had type 2 diabetes without nephropathy and 20 healthy subjects. RESULTS: In the diabetic patients with poor glycemic control, urinary sorbitol levels were significantly increased and urinary myo-inositol excretion was approximately 6.5-fold higher than in healthy controls (33.0+/-6.5 vs 221.7+/-45.9 mg/day, mean+/-SE, P<0.01). During strict glycemic control, some patients (Group A) showed simultaneous disappearance of glucosuria and normalization of the urinary excretion of myo-inositol (<50 mg/day) and, while others (Group B) showed delayed normalization of urinary myo-inositol excretion. Group B showed significantly higher urinary myo-inositol, sorbitol, and fructose excretion than Group A at the time of disappearance of glucosuria. These findings suggest that patients in Group B may have increased polyol pathway activity. CONCLUSION: Even though short-term strict glycemic regulations were established in long-standing hyperglycemic diabetic patients, to normalize the once-exaggerated polyol pathway activities, it was essential to maintain glucosuria-free conditions for some period. Quantitation of urinary polyols using GC/MS appears to be a clinically useful method for assessing polyol pathway activity.

aFGF, endogenous satiety substance, facilitates learning, memory and immune function in aging.

aFGF injection s.c. once a week into SAMP8 was begun at 3 weeks after birth and continued for 10 months. Saline was injected as a control. learning and memory and cellular immunological functions in the aFGF group were enhanced significantly, while those of the saline group deteriorated. 1. The number of cholinergic neurons was decreased by less than 20% and ChAT activity in individual neurons in the medial septum which send monosyonaptic terminals to the hippocampus was significantly decreased in the saline group, but not so much in the aFGF group. 2. The respective densities of muscarinic and aFGF receptors, on the hippocampal neurons were significantly higher in the aFGF group than in the saline group. 3. The LTP in hippocampal slice preparations was significantly facilitated in the aFGF group, but not in the saline group. 4. The DTH, (T cell immune response) measured at the end of the 2nd and 7th months were reduced in the 7th month as compared with the 2nd month in the saline group, but aFGF group protected against this reduction. 5. These results show that aFGF provides protection against impairment of not only learning and memory but also the DTH immunoreactivity in SAMP8.

[Three children with systemic cat scratch disease].

Three girls with systemic cat scratch disease, aged 10, 13 and 9 years, were reported. They presented a prolonged fever and back pain in the early stage of the disease, and had no regional lymphadenopathy. Two of them had hepatosplenic granulomas, one with multiple 5 mm hypoechoic lesions in the liver and spleen, and the other with a single 2.5 cm hypodense lesion in the left hepatic lobe. The latter patient underwent a partial left hepatic lobectomy. All patients had elevated titers of antibodies to Bartonella henselae. Polymerase chain reaction detected B. henselae DNA in tissue specimens of the patient who underwent a hepatic lobectomy. Cat scratch disease should be recognized as a cause of fever of unknown origin because the prevalence of B henselae infection might be higher in Japan.

[Inhibitory effect of histamine-added mouse gamma-globulin on eosinophil accumulation induced by allergen in BALB/c mice].

Histamine-added human gamma-globulin (HG) has been clinically used as an anti-allergic drug for asthma, allergic rhinitis and atopic dermatitis. Retrospective analysis of clinical data have indicated that s.c. administration with HG not only improves clinical symptoms but also suppresses the number of eosinophils of nasal secretion or peripheral blood in allergic patients. Thus, the possibility was explored that HG may actively suppress eosinophil accumulation in allergic inflammation. The eosinophil accumulation in peritoneal cavity was induced by i.p. injection with ragweed pollen extract in BALB/c mice which had been repeatedly sensitized with the allergen for three weeks. Histamine-added mouse gamma-globulin (Mouse HG) at 150 mg/kg/day markedly inhibited the allergen-induced eosinophil accumulation when administered s.c. two times a week for three weeks. The inhibitory effect was almost the same as that of cyclosporin A at 100 mg/kg/day. Interestingly, equivalent dose of histamine or mouse gamma-globulin alone had no inhibitory effect in the same system. These results suggest that HG suppresses chronic allergic inflammation through the inhibition of eosinophil accumulation.

[Measurement of HbA1c as a method to diagnose diabetes mellitus in nursing home residents].

Measurement of HbA1c is widely used to control diabetes mellitus (DM) and is one of the methods to diagnose DM. We adopted the measurement of HBA1c in the annual medical examination of residents at a nursing home for the elderly in order to obtain the normal serum level of HbA1c in the elderly and analyze the influence of aging on HbA1c. HbA1c and the body mass index (BMI) were measured in 968 elderly. HbA1c was distributed from 3.7 to 12.5%. Mean and SD, were 5.03 +/- 0.61% and there was no difference of the serum level of HbA1c between males and females. HbA1c was below 6.0% in 95% of all subjects. There was no correlation between age and HbA1c. To determine data trends, we divided the elderly into 4 age groups of 60 to 69 years, 70 to 79 years, 80 to 89 years and 90 or more. With advancing age, HbA1c gradually increased but there was no significant difference of HbA1c among the 3 groups which were younger than the group aged 90 or more. HbA1c was 5.51 +/- 1.06% in the group aged more than 90 years and there was a significant difference in the value of HbA1c between the group aged 90 or more and other groups. Inversely, BMI gradually decreased with age and there was also a significant difference of BMI between the group aged 90 or more and the other groups. These results suggest that there is a difference of metabolic condition between the group aged 90 or more and the other groups.(ABSTRACT TRUNCATED AT 250 WORDS)

[Hemodynamic changes in acute pancreatitis].

Hemodynamics studies were carried out from days 1-5 following the onset of illness in 7 patients with severe pancreatitis (group A) and in 7 patients with moderate pancreatitis (group B). Patients in both groups had a higher cardiac index (CI) and a lower systemic vascular resistance (SVR) than normal patients during 5 days of illness, and patients in group A had a higher CI of 5.38 +/- 0.841 x min/m2 and lower SVR of 889 +/- 234dyn.sec/cm5 than those in group B on day 4. Patients in group A had a lower pulmonary vascular resistance than normal patients on days 1, 3, and 4, but those in group B did not show this hemodynamic change. Group A patients had a higher pulmonary wedge pressure of 11.9 +/- 8.4mmHg and depressed left ventricular stroke work index of 59.8 +/- 17.8g.m/m2 as compared with group B (5.6 +/- 3.4mmHg, 77.7 +/- 23.6g./m2, respectively). These findings indicate that a hyperdynamic hemodynamic state may exist in the early stages of moderate and severe pancreatitis and myocardial depression may be evident in severe pancreatitis.