Comparison of twice-daily injections of biphasic insulin lispro and basal-bolus therapy: glycaemic control and quality-of-life of insulin-naïve type 2 diabetic patients.

OBJECTIVE: The aim of this study was to evaluate twice-daily injections of biphasic insulin lispro vs. basal-bolus (BB) therapy with regard to quality-of-life (QOL) and glycaemic control in insulin-naïve type 2 diabetic patients. METHODS: Twenty-eight patients with type 2 diabetes were randomized to receive either twice-daily 50/50 premixed insulin lispro (Mix50 group) or BB (NPH insulin at bedtime and preprandial insulin lispro) therapy (BB group) for 12 weeks. Glycated haemoglobin (HbA1C), 1,5-anhydroglucitol (1,5-AG), blood plasma glucose level, body mass index (BMI), daily total insulin dosage and insulin therapy-related QOL (ITR-QOL) were studied. RESULTS: ITR-QOL was significantly better in the Mix50 than in the BB group (103.1 +/- 9.8 vs. 90.6 +/- 19.4; p < 0.05). HbA(1c) improved in both groups (from 11.1 +/- 2.1 to 6.9 +/- 1.0% with Mix50 vs. from 11.0 +/- 2.3 to 6.6 +/- 0.8% with BB therapy). CONCLUSION: These results might suggest that twice-daily injections of premixed rapid-acting insulin analogue therapy could achieve good glycaemic control and better QOL compared with BB therapy in insulin-naïve type 2 diabetes.

A case of IgG4-related autoimmune disease with multiple organ involvement.

A 52-year-old male was admitted with autoimmune pancreatitis (AIP), showing mononuclear cell infiltration in both the pancreas and salivary glands with both normal sialography and anti-SS-A/SS-B antibodies. Although the AIP improved with glucocorticoid treatment, subsequent abdominal computed tomography (CT) revealed a nodular shadow in the bilateral kidneys, which was confirmed as interstitial nephritis by renal biopsy. The patient's serum immunoglobulin G4 (IgG4) level was 10 times higher than the upper limit of the normal range. IgG4-positive mononuclear cell infiltration was detected in the salivary gland, pancreas, and kidney. A new entity proposed as 'IgG4-related autoimmune disease' was considered.

N-acetyl-cysteine accelerates transfer of diabetes into non-obese diabetic scid mice.

AIMS/HYPOTHESIS: Type 1 diabetes mellitus is caused by autoimmune pancreatic beta cell destruction, and the destructive process involves several molecular mechanisms including oxygen-reactive species. A cysteine derivative, N-acetyl-cysteine, is widely used as an antioxidant, but the role of N-acetyl-cysteine in the protection of pancreatic beta cells in type 1 diabetes remains unclear. The aim of this study was to clarify the effect of N-acetyl-cysteine on beta cells using an adoptive transfer system in a murine model of type 1 diabetes. METHODS: Splenocytes from diabetic female non-obese diabetic mice were transferred into female non-obese diabetic scid/ scid recipients to induce diabetes. Just after transfer, N-acetyl-cysteine was administered to non-obese diabetic scid recipients. Two weeks after transfer, the pancreas of the recipients was examined histologically, and cytokine mRNA expression in the pancreas was analysed. In vitro, CD4-positive splenocytes from diabetic donor mice were stimulated with anti-CD3 and anti-CD28 antibodies with or without N-acetyl-cysteine. RESULTS: Treatment with N-acetyl-cysteine significantly accelerated the transfer of diabetes into non-obese diabetic scid recipients. Treatment with N-acetyl-cysteine accelerated the infiltration of mononuclear cells accompanied by CD8-positive cells into the intra-islet region of the recipient's pancreas, and enhanced interferon-gamma mRNA expression in the pancreas. In vitro, treatment with N-acetyl-cysteine enhanced interferon-gamma and interleukin-2 production by CD4-positive splenocytes of the diabetic donor mice. CONCLUSIONS/INTERPRETATION: N-acetyl-cysteine accelerates the transfer of diabetes into non-obese diabetic scid mice and this effect is accompanied by the promotion of local infiltration and T-helper cell type 1 responses.

KF31327, a new potent and selective inhibitor of cyclic nucleotide phosphodiesterase 5.

The effects of KF31327 (3-ethyl-8-[2-(4-hydroxymethylpiperidino)benzylamino]-2,3-dihydro-1H-imidazo[4,5-g]quinazoline-2-thione dihydrochloride) on phosphodiesterase 5 (cyclic GMP-specific phosphodiesterase) activity and platelet aggregation were investigated and compared with those of sildenafil, a well-known phosphodiesterase 5 inhibitor. KF31327 inhibited phosphodiesterase 5 from canine trachea (K(i)=0.16 nM) more potently than sildenafil (K(i)=7.2 nM). The kinetic analysis revealed that KF31327 was a non-competitive inhibitor. In the presence of nitroglycerin (nitric oxide generator), both compounds inhibited the collagen-induced aggregation of rabbit platelets at less than 0.1 microM, augmenting intracellular cyclic GMP level without affecting cyclic AMP. In contrast, in the absence of nitroglycerin, a higher concentration (10 microM) of KF31327 was required to inhibit platelet aggregation and increased both cyclic nucleotide levels. However, 10 microM sildenafil did not affect aggregation despite elevation of cyclic GMP comparable to that in the presence of nitroglycerin. These results indicate that in the presence of nitroglycerin, the inhibition of platelet aggregation by KF31327 is due to the elevation of cyclic GMP, whereas the mechanism underlying the inhibition without nitroglycerin might be related to a rise in intracellular cyclic AMP.

New insights into the transmembrane protein tyrosine phosphatase CD45.

CD45 is expressed on all nucleated haematopoietic cells and was originally identified as the first and prototypic transmembrane protein tyrosine phosphatase. In CD45 mutant cell lines, CD45-deficient mice and CD45-deficient human SCID patients, CD45 is required for signal transduction through antigen receptors. CD45 can operate as a positive as well as a negative regulator of Src-family kinases. Moreover, CD45 was identified as the elusive JAK tyrosine phosphatase that negatively regulates cytokine receptor activation involved in the differentiation, proliferation and antiviral immunity of haematopoietic cells. Modulation of CD45 splice variants provides a unique opportunity to design drugs that turn off or turn on antigen and cytokine receptor signaling in cancer, transplantation or autoimmunity

[The effects of milrinone on hemodynamics in patients undergoing cardiac surgery].

The phosphodiesterase inhibitor, milrinone is used to treat low cardiac output syndrome, especially after cardiac surgery. But there were few reports about the precise hemodynamic effects at separation from cardiopulmonary bypass (CPB). We examined the hemodynamic effects of milrinone in 24 patients undergoing elective coronary artery bypass graft (CABG). Patients were assigned to the milrinone group (n = 12) and the control group (n = 12). Before separation from CPB, milrinone was administered as a loading dose of 50 micrograms.kg-1 into the reservoir of CPB at rectal temperature 33.5 degrees C and simultaneously a continuous infusion of 0.5 microgram.kg-1.min-1 was started. In addition, dopamine and nitroglycerine were administered in both groups. Hemodynamic measurements were performed before CPB, just after the weaning from CPB, 15, 30, 60 minutes after the weaning from CPB. Cardiac index increased significantly (P < 0.01) in the milrinone group as compared with the control group. Systemic vascular resistance index and mean arterial pressure decreased significantly (P < 0.0001, P < 0.05, respectively) in the milrinone group as compared with the control group. There were no significant differences in heart rate, mean pulmonary arterial pressure, pulmonary artery occlusion pressure, mean right atrial pressure, stroke volume index, and pulmonary vascular resistance index between the two groups. These hemodynamic effects showed that milrinone supported cardiac performance after CPB for CABG.

Relationship between flow cytometric DNA ploidy and nuclear grade with endocrine dysfunction in adrenal cortical adenomas.

OBJECTIVES: To determine the relationship among the DNA ploidy, histopathologic features, and clinical syndrome in adrenal cortical adenomas, because the cells often show variability in nuclear size and configuration. METHODS: Our study included 44 adenomas associated with primary aldosteronism and 23 adenomas associated with Cushing syndrome. Normal adrenal glands from patients with renal carcinoma served as the controls. Paraffin-embedded tissues were examined for DNA content by flow cytometry. The mean percentage of G(2)/M (4C%) of the control samples was 3.8%. Tetraploid was represented by a histogram with both a 4C peak greater than 9% (mean + 2.4 SD of control samples) and a small 8C peak. RESULTS: Flow cytometric analysis revealed diploidy in 30, tetraploidy in 27, and aneuploidy in 8 of the 67 adenomas; 2 adenomas could not be classified. All 17 normal adrenal glands showed diploidy. A significant relationship was noted between DNA ploidy and the clinical syndrome (ie, a larger proportion of adenomas with primary aldosteronism had a tetraploid DNA histogram compared with adenomas with Cushing syndrome, P <0. 0001). Adenomas with primary aldosteronism had a significantly higher nuclear grade (III or IV) than did tumors with Cushing syndrome (P = 0.033). A significant relationship was also observed between DNA ploidy and nuclear grade in 57 euploid tumors, with tetraploid tumors often showing the highest nuclear grade (P = 0. 037). CONCLUSIONS: Our results have demonstrated that adrenal cortical adenomas associated with primary aldosteronism often reveal severe nuclear pleomorphism, indicating that nuclear pleomorphism might be due to a tetraploid stemline.

[X-Linked Charcot-Marie-Tooth disease with a new mutation (Thr191Ala) in the connexin32].

We report a 59-year-old man with X-linked Charcot-Marie-Tooth (CMT) disease and a new point mutation in the connexin32 gene. The patient first noticed mild gait disturbance five years previously. On admission, he exhibited muscle atrophy and weakness in the distal part of both legs, mild muscle atrophy of both hands without weakness, and a minimal reduction of touch sensation in the right dorsal foot. Nerve conduction velocity of the peripheral nerves was diffusely reduced. Electromyography exhibited high-amplitude, long-duration, polyphasic motor unit potentials in the muscles of the extremities. Fibrillation potential and positive sharp wave were present in the affected muscles. Cerebrospinal fluid protein was slightly elevated. The polyneuropathy did not respond to high-dose corticosteroid treatment, and showed very slow progression. His parents were not consanguineous. His father and two sons were healthy, but similar illness (more severe) was suspected in his younger brother. Gene analysis (Southern hybridization) did not reveal any duplication or deletion in the CMT 1 A-REP region. However, a novel mutation (Thr191Ala) was detected in the connexin32. Although more than 160 mutations in the connexin 32 gene have been identified worldwide, approximately ten mutations have so far been reported in Japan. In comparison with X-linked CMT patients with other connexin32 mutations, the present case was characterized by late onset and mild neurological manifestation. Gene analysis provides a useful tool for diagnosing cases with slowly progressive, motor dominant polyneuropathy of unknown origin.

Structural and functional abnormalities in the spleen of an mFtz-F1 gene-disrupted mouse.

The spleen has two main functions. The first is to provide a proper microenvironment to lymphoid and myeloid cells, whereas the second involves clearance of abnormal erythrocytes. Ad4BP/SF-1, a product of the mammalian FTZ-F1 gene (mFTZ-F1), was originally identified as a steroidogenic, tissue-specific transcription factor. Immunohistochemical examination of the mammalian spleens confirmed the expression of Ad4BP/SF-1 in endothelial cells of the splenic venous sinuses and pulp vein. In mFtz-F1 gene-disrupted (KO) mice, several structural abnormalities were detected in the spleen, including underdevelopment and nonuniform distribution of erythrocytes. Examination of the spleen of KO fetuses showed failure of development of certain tubular structures during embryogenesis. These structures are normally assembled by Ad4BP/SF-1 immunoreactive cells, and most likely form the vascular system during later stages of development. Other structural abnormalities in the spleen of the KO mice included defects in the tissue distribution of type-IV collagen, laminin, c-kit, and vimentin. These morphologic defects in the vascular system were associated with a decrease in the proportion of hematopoietic cells, although differentiation of these cells was not affected significantly. A high number of abnormal red blood cells containing Howell-Jolly bodies were noted in the KO mice, indicating impaired clearance by the splenic vascular system. We also detected the presence of an mRNA-encoding cholesterol side-chain cleavage P450 in the spleen, resembling the findings in steroidogenic tissues such as the gonads and adrenal cortex. The mRNA transcript was not involved in splenic structural defects as it was detected in the spleens of both normal and KO mice, indicating that the regulatory mechanism of the P450 gene in the spleen is different from that in steroidogenic tissues. Our results indicate that a lack of the mFtz-F1 gene in mice is associated with structural and functional abnormalities of the splenic vascular system.

Effect of triiodothyronine on muscle cell differentiation and blood glucose level in hyperglycemic KK mice.

KK mice are genetically diabetic animals, showing glucose intolerance and insulin resistance. We examined the effects of 3,3',5-triiodo-L-thyronine (T3) on the blood glucose level and on mRNA levels of muscle cell differentiation markers in hyperglycemic KK mice. T3 treatment (T1, 1 mg; T3, 3 mg; T10, 10 mg/kg/day) of KK mice for 4 days caused a decrease in blood glucose level by 11%, 25%, and 24%, respectively, without affecting body weight. Skeletal muscle of mice treated with T3 (T10) showed a 98% increase in the mRNA level of the glucose transporter isotype 4 (Glut4). In contrast, T3 treatment did not affect the mRNA level of the isotype 1 (Glut1) transporter. The mRNA level of a muscle cell specific differentiation marker, MyoD, showed a significant increase in the T3 treatment group with an accompanying enhancement of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA level. These results suggest that T3 stimulates muscle cell differentiation in vivo, concomitant with a stimulation of cellular glucose metabolism, thus decreasing the blood glucose level in hyperglycemic KK mice.

Immunohistochemical detection of nm23-H1/NDP kinase in childhood thyroid carcinoma.

Childhood thyroid cancer is known to be aggressive. High incidence of lymph node and distant metastasis are characteristic features of these cases. In adult, reduced expression of nm23-H1/nucleoside diphosphate (NDP) kinase has been correlated with cancer invasion and metastasis in some tumor types. Therefore, we examined the expression of nm23-H1 gene product in childhood thyroid carcinomas in Japan. 27 primary thyroid carcinomas and 8 metastatic lymph nodes were analyzed by immunohistochemistry using monoclonal antibody H1-229. 21 out of 23 cases (91%) of papillary carcinomas were positively immunostained, whereas none of the 4 follicular carcinomas showed any immunoreactivity. No correlation was found between the nm23-H1/NDP kinase antigen expression and nodal involvement or distant metastasis in primary tumors. However, only 50% (4 out of 8) of metastatic lymph nodes from papillary carcinoma were positively stained, demonstrating a significant decrease comparing to those of primary sites. These data indicate that the expression of nm23-H1/NDP kinase cannot predict tumor metastatic potential in childhood thyroid cancer.

Glycerol-3-phosphate Dehydrogenase Inhibitors, Anacardic Acids, from Ginkgo biloba.

Four inhibitors of glycerol-3-phosphate dehydrogenase were isolated from Ginkgo biloba and identified as anacardic acids (6-tridecylsalicylic acid, 6-[(8Z)-pentadecenyl]salicylic acid, 6-[(9Z,12Z)-heptadecadienyl]salicylic acid, and 6-[(8Z)-heptadecenyl]salicylic acid) by instrumental analyses. Their 50% inhibitory concentrations against the enzyme were 1-3 µg/ml under the standard assay conditions. Anacardic acid inhibited the enzyme non-competitively. The sarcotesta contained most of anacardic acids, and nuts a little.

Proliferating cell nuclear antigen in ACTH-independent bilateral macronodular adrenal hyperplasia.

ACTH-independent bilateral macronodular adrenal hyperplasia (AIMAH) has been recently described as a rare cause of Cushing's syndrome and it is an unusual process of unknown etiology. This report describes a case of a 59-year-old man with AIMAH. The proliferating cell nuclear antigen (PCNA) has been implicated in the process of cell proliferation and is detectable throughout most of the cell cycle. This report also describes the expression of PCNA in AIMAH by immunohistochemical staining. Most of the positive expression was seen in the greater part of the epithelial cells of the cortical lesion, but not in the interstitial cells. These data suggest that some effective factor, specific for adrenal cortical cell growth, might be produced in AIMAH.

Adrenocorticotropic hormone-independent bilateral macronodular adrenocortical hyperplasia associated with Cushing's syndrome.

A case of adrenocorticotropic hormone independent bilateral adrenocortical macronodular hyperplasia (AIMAH) is reported. A 59 year old male was admitted to hospital because of hypertension. Subsequently, hypercortisolism, low plasma adrenocorticotropic hormone (ACTH), loss of diurnal rhythm of ACTH, lack of suppression with high dose dexamethasone were found and bilateral adrenal enlargement was detected by abdominal computerized tomography and adrenal scintigraphy. Bilateral total adrenalectomy was performed under a diagnosis of bilateral adrenal hyperplasia associated with Cushing's syndrome. Both adrenal glands were enlarged in size and weight. Bulging nodules were found at the cut section. Microscopically, a variegated histologic pattern including trabecular, adenoid and zona glomerulosa-like (ZG-like) structures was revealed in the nodules. Immunohistochemical examination disclosed positive staining of cytochrome P-450 17 alpha, negative of 3 beta-HSD in the ZG-like structure. Ultrastructurally, the cells composing the ZG-like structure were similar to those of the ZG in normal adrenal cortex. The authors agree that AIMAH is one of the entities causing Cushing's syndrome, and advise pathologists to keep this disorder in mind when they examine the adrenals in Cushing's syndrome.

Cloning and sequencing of a Synechococcus gene encoding a protein very similar to mammalian aldehyde dehydrogenases.

We report cloning and sequencing of a gene encoding a putative aldehyde dehydrogenase in Synechococcus sp. PCC7942. It was found that this phototrophic microorganism has a protein very similar to mammalian class-3 aldehyde dehydrogenases. A mutant strain lacking this gene was hypersensitive in growth to an aromatic aldehyde.

Case report: ciliated foregut cyst of the pancreas mimicking teratomatous tumour.

A ciliated cyst of foregut origin is often found in relation to the tracheobronchial tree and the upper gastrointestinal tract, when it is called bronchogenic cyst and enterogenous cyst, respectively. Cysts of this type rarely occur in the pancreas. We report a case of ciliated foregut cyst involving the pancreas, and mimicking teratomatous tumour on radiology.

Functional-morphological effect of the synthetic atrial natriuretic factor (ANF 101-126) on the zona glomerulosa cells of adrenal cortex.

The effects of atrial natriuretic factor (ANF) on the adrenal steroidogenesis were examined in the zona glomerulosa of rats in vivo. ANF administration provoked a significant decrease in the width of zona glomerulosa (P < 0.01) and a significant enlargement of nuclei of zona glomerulosa cells (P < 0.01). Ultrastructurally conspicuous change was observed in mitochondria of the cells of the zona glomerulosa treated with ANF, representing elongated mitochondria with "parallel arrays" of tubules with pipe-like structure. In vitro autoradiography 125I-labelled ANF binding displayed a significant decrease of a specific receptor site in the zona glomerulosa treated with ANF. A significant lowering in plasma levels of aldosterone was observed in ANF-treated rats. These findings could be interpreted as an inhibitory effect of ANF on adrenal steroidogenesis of the glomerular zone by action through a specific ANF-receptor.