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Chronic nonbacterial osteomyelitis is a rare bone disorder that can be found in the jaw. It is often associated with systemic conditions, including autoimmune deficiencies. However, little is known about how the genetic and immunologic background of patients influences the disease. Here, we focus on human leukocyte antigen (HLA), killer cell immunoglobulin-like receptors (KIRs), and their specific combinations that have been difficult to analyze owing to their high diversity. We employed a recently developed technology of simultaneous typing of HLA alleles and KIR haplotype and investigated alleles of the 35 HLA loci and KIR haplotypes composed of centromeric and telomeric motifs in 18 cases and 18 controls for discovery and 472 independent controls for validation. We identified an amino acid substitution of threonine at position 94 of HLA-C in combination with the telomeric KIR genotype of haplotype tA01/tB01 that had significantly higher frequency (>20%) in the case population than in both control populations. Multiple logistic regression analysis based on a dominant model with adjustments for age and sex revealed and validated its statistical significance and high predictive accuracy (C-statistic ≥0.85). Structure-based analysis revealed that the combination of the amino acid change in HLA-C and the telomeric genotype tA01/tB01 could be associated with lower stability of HLA-C. This is the first case-control study of a rare disease that employed the latest sequencing technology enabling simultaneous typing and investigated amino acid polymorphisms at HLA loci in combination with KIR haplotype.

BackgroundElevated serum IgG4 levels are one of the characteristic findings in immunoglobulin G4 (IgG4)-related disease (IgG4-RD). Serum IgG4 levels have an impact to a certain extent on the diagnosis of IgG4-RD although there are some issues in their sensitivity and specificity. In the reports from Japan, China, USA, and Europe, elevated serum IgG4 levels were reported to be observed in 83-97% of patients with IgG4-RD [1-5]. In the past investigations of hospital patients, some studies reported that 10-15% of hospital patients with elevated serum IgG4 levels had IgG4-RD [6,7]. However, in general adults with no symptom, investigations of prevalence of elevated serum IgG4 levels and/or IgG4-RD have rarely been conducted.ObjectivesThis study aimed to investigate the frequency of serum IgG4 elevation in the general Japanese population and its associated factors using data from resident examinations.MethodsWe measured the serum IgG4 levels in 1,204 residents who underwent a general medical examination in Ishikawa prefecture, Japan. Logistic regression analysis was used to search for factors related to elevated serum IgG4 levels. Secondary examinations were conducted for participants in whom elevation was identified.ResultsThe mean serum IgG4 level was 44 mg/dL, and elevated serum IgG4 levels were observed in 42 patients (3.5%). Univariate logistic regression analyses showed that male sex, older age, lower estimated glomerular filtration rates based on cystatin C (eGFR-CysC), serum high-density lipoprotein cholesterol levels, and higher hemoglobin A1c (HbA1c) levels were associated with elevated serum IgG4 levels. Subgroup analyses in men showed that older age, lower eGFR-CysC levels, and higher serum HbA1c levels were associated with elevated serum IgG4 levels. In contrast, the analyses in women found no significant factors. One of the 10 residents who underwent secondary examinations was diagnosed with possible IgG4-related retroperitoneal fibrosis.ConclusionIn the general population, elevated serum IgG4 levels are more common in elderly men, which is similar to the epidemiological features of IgG4-RD.References[1]Inoue D, et al. IgG4-related disease: dataset of 235 consecutive patients. Medicine (Baltimore). 2015;94(15):e680.[2]Yamada K, et al. New clues to the nature of immunoglobulin G4-related disease: a retrospective Japanese multicenter study of baseline clinical features of 334 cases. Arthritis Res Ther. 2017;19(1):262[3]Culver EL, et al. Elevated serum IgG4 levels in diagnosis, treatment response, organ involvement, and relapse in a prospective IgG4-related disease UK cohort. Am J Gastroenterol 2016;111:733–43.[4]Lin W, et al. Clinical characteristics of immunoglobulin G4-related disease: a prospective study of 118 Chinese patients. Rheumatology (Oxford). 2015;54(11):1982–90.[5]Carruthers MN, et al. The diagnostic utility of serum IgG4 concentrations in IgG4-related disease. Ann Rheum Dis 2015;74:14-18.[6]James Yun, et al. Poor positive predictive value of serum immunoglobulin G4 concentrations in the diagnosis of immunoglobulin G4-related sclerosing disease. Asia Pac Allergy. 2014 Jul;4(3):172-176.[7]Taiwo N Ngwa, et al. Sreum immunoglobulin G4 ｌevel is a poor predictor of immunoglobulin G4–related disease. Pancreas. 2014 Jul;43(5):704-7.Disclosure of InterestsNone declared

Supplemental material, DS_10.1177_0022034520901519 for A Targeted Genetic Association Study of the Rare Type of Osteomyelitis by H. Yahara, S. Horita, S. Yanamoto, Y. Kitagawa, T. Asaka, T. Yoda, K. Morita, Y. Michi, M. Takechi, H. Shimasue, Y. Maruoka, E. Kondo, J. Kusukawa, H. Tsujiguchi, T. Sato, T. Kannon, H. Nakamura, A. Tajima, K. Hosomichi and K. Yahara in Journal of Dental Research

We have already reported a prediction method of flux losses (FLs) in bonded magnets and showed good agreement between the calculated losses and the measured ones. In this paper, we improve our previously reported method, and investigate a FL of a Nd–Fe–B-bonded magnet under external magnetic fields. To verify the validity of the prediction method for the FL, we evaluated and calculated a FL of a cylindrical Nd–Fe–B-bonded magnet under the external magnetic fields of −240, −480, and −720 kA/m. The FL of the magnet exposed at 120 °C depended on the strength of the external field, and the measured values of the loss were 8.79% for −240 kA/m, 61.2% for −480 kA/m, and 149.9% for −720 kA/m. The calculated values were 6.44%, 61.1%, and 141%, and we obtained good agreement between the measured and calculated ones. From the results, we conclude that our improved method is effective to predict the FL of the Nd–Fe–B-bonded magnets under external magnetic fields.

A porous alumina body was synthesized from anisotropic alumina particles (platelets). The uniaxial pressure in fabricating the green compact body had an influence on the relative density of the alumina body after heating. When green compacts, which had been uniaxially pressed at 1 and 3 MPa, were heated at 1400 °C for 1 h, the relative densities of the resulting alumina bodies were 25.0% and 35.5%, respectively. The compressive strength of compacts that were uniaxially pressed at 1 and 3 MPa were 0.8 and 4.3 MPa, respectively. In an attempt to increase the compressive strength of these porous alumina bodies, aluminum nitrate and magnesium nitrate solution treatments were performed, followed by reheating to 1400 °C for 1 h. When a 0.5 mol/l aluminum nitrate solution was used, the compressive strength of the porous alumina body uniaxially pressed at 1 MPa changed from 0.8 MPa (without solution treatment) to 1.5 MPa. Furthermore, when 0.1 mol/l magnesium nitrate solution was used, the compressive strength of the porous alumina increased to 1.7 MPa. Thus, solution treatment of the porous alumina body had a strong positive effect on its mechanical strength.

High resolution X-ray structure analyses and electronic structure calculation revealed the condition for the charge ordering(CO) observed in the (TMTTF) 2 X (Fabre) salts. The phase diagram of the electronic states including 2k F and 4k F CO has been proposed associated with magnitudes of molecular dimerization and tetramerization.

The aim of this study was to clarify the histopathologic significance of allograft glomerulitis in chronic allograft nephropathy (CAN). Review of our renal allograft biopsy files revealed 140 specimens with CAN among 115 selected patients. They were classified into two groups: one had CAN with glomerulitis (group G), and the other was free of this finding (group NG). We evaluated the clinicopathologic parameters as follows: levels of serum creatinine and proteinuria in the biopsy; presence of circulating anti-donor antibodies; allograft failure rate; history of biopsy-proven acute cellular rejection (ACR) and acute humoral rejection (AHR); complications of ACR and chronic rejection (CR); and results of immunofluorescence studies for C4d and HLA-DR. The glomerulitis group showed a significantly greater incidence of CR complications, the presence of circulating anti-donor antibodies, and C4d deposition in peritubular and glomerular capillaries. This group also showed higher levels of serum creatinine and proteinuria, higher graft loss rate, and increased AHR incidence, although the differences were not significant. There was also no statistical significance in the HLA-DR expression on tubular epithelial cells. The present results strongly suggest that humoral factors may play an important role in the progression of glomerulitis in CAN. Therefore, we suspect that glomerulitis in CAN is one of the main histologic markers for CR. The presence of glomerulitis may represent humoral factor-dependent inflammation. It should be considered an important diagnostic criterion for CR in addition to double-contour formation and elastica disruptions with or without subendothelial inflammation (Banff '97).

The extent of the oxidation of Hemoglobin (Hb) M Saskatoon (beta 63His--Tyr) and Hb M Boston (alpha 58His--Tyr) in the patient's blood was determined by measurement of the intensity of EPR signals at g perpendicular = 6.0 for the normal subunits, g1 = 6.7 for the mutant subunits of Hb M Saskatoon and g1 = 6.3 for those of Hb M Boston, respectively. The amounts of reduced mutant subunits were estimated from the EPR signal intensities and the amounts of Hb present as mutant Hb in the blood. About 50% and 76% of mutant subunits in Hb M Boston and Hb M Saskatoon remained reduced in the fresh blood. Gentle shaking of the blood at 37 degrees C for 15 hours in air brought about autoxidation of the normal subunits as well as the mutant subunits of the two Hbs M, indicating that the presence of the mutant subunits facilitated autoxidation of the normal subunits. Possible involvement of NADH-metHb reductase in erythrocytes in maintenance of the reduced mutant subunits of Hb M Saskatoon was discussed.

The chemical composition of sputter-Deposited BiSrCaCuO thin films was investigated as a function of system geometry and some controllable parameters. Films prepared using conventional configuration were bismuth deficient owing to the selective resputtering by energetic oxygen particles released at the target surface. Exploiting an unusual configuration, which included both an off-axial substrate position and a grounded plate in front of the target, the film stoichiometry reproduced that of the target. The fundamental properties of prepared films are also considered in the paper. In particular, the crystal structure and the superconductivity of in-situ grown stoichiometric films are discussed.

Alpha-fetoprotein (AFP) is an important tumor marker for yolk sac tumor and hepatoblastoma in childhood. We have been using the graph of the normal range of serum AFP made by Tsuchida et al, when we evaluate the serum AFP levels in early infancy. We measured the serum AFP levels by an immunoradiometric assay in 163 normal infants under 2 years of age, in order to make a more precise graph. Our normal range was a little wider than that of Tsuchida et al. According to our graph, false-positive cases would be fewer. Referring to the half-lives of serum AFP levels in normal infancy is also useful, when it is difficult to evaluate the AFP level.

To investigate the expression of the cadherin complex in human crescentic glomerulonephritis to elucidate the role of intercellular adherens junction molecules in crescent formation.Immunostaining revealed cadherin complexes localized in Bowman's epithelial cells, but not in podocytes, of normal human glomeruli. Eight adult cases with myeloperoxidase anti-neutrophil cytoplasmic autoantibodies (MPO-ANCA)-related (pauci-immune type) crescentic glomerulonephritis were examined on immunofluorescence microscopy with anti-pan cadherin, p120 catenin, and beta-catenin antibodies. The specimens provided six cellular crescents, 12 fibrocellular crescents, and four fibrotic crescents. Immunofluorescence was semiquantitatively estimated by the rate of the field of localization within the whole area of the crescent, according to the four-grade system [(-) - (++)]. All the tested molecules consisting of the cadherin complex were abundantly observed in cytokeratin-positive epithelial components in crescents, each with an equivalent area of localization. The expression of the cadherin complex was closely associated with that of cytokeratin and both diminished as the crescents developed from cellular to fibrotic.The cadherin-catenin complex is a specific marker of Bowman's epithelial cells in human glomeruli. The cellular crescents in pauci-immune-type crescentic glomerulonephritis possess adherens junction molecules, indicating a principle parietal epithelial cell phenotype.

Super-resolution direction finding algorithms such as MUSIC or ESPRIT methods inherently assume narrow-band signals, which are not applicable to ultra-wideband radars such as GPR (ground penetration radar). We developed a time-domain algorithm to determine the time and direction of arrival pulses with an order higher resolutions than the nominal resolution determined by the aperture size. The received signal waveform at each array element is first analyzed by the recursive nonorthogonal decomposition using a dictionary of basic waveforms. After the delay time of the echoes in each element of the receiving array is determined, the number of targets and their locations are determined by analyzing variation of the delay time over the array with the aid of a Hough transform. The robustness of the proposed algorithm is confirmed by numerical simulations.

A 38-yr-old man with factor H dysfunction and unknown glomerular disease received first and second renal transplantations (Tx) from living-related donors. His examination showed a low percentage activity of factor H (31%). Factor H dysfunction has been known to be associated with type II or III membranoproliferative glomerulonephritis (MPGN), haemolytic uraemic syndrome and IgA GN. The first graft from his mother showed diffuse mesangial deposit of IgA. His son has had IgA GN and his data also revealed a low percentage activity of factor H (33%). He and his son both showed a low activity of C3. Moreover, his father, who was the donor of the second Tx, had a low percentage activity of factor H (25%), and presented with mild glomerular deposit of C3 at operation, while he has been healthy through his entire 67 yr of life. Each of them had a low percentage activity of factor H. These findings through three generations suggested the inheritance of factor H dysfunction. The patient presented with proteinuria 3 months after the first Tx. At the first biopsy 30 months after the first Tx, light microscopy revealed minor glomerular abnormalities with electron dense deposits in subepithelial, intramembranous and mesangial regions, while immunofluorescence showed massive glomerular deposits of C3. In the second biopsy 51 months after the first Tx, the glomerulonephritis developed mesangial proliferation and crescent formation, accompanied by more massive C3 deposit and intramembranous, mesangial and subepithelial dense deposits. He then required redialysis. At the second and third biopsies within 2 months after the second Tx, the renal graft showed similar findings to the first biopsy after the first Tx. He perhaps presented with a recurrence of complement-associated GN, showing an atypical form of MPGN after Tx. These findings suggest that factor H dysfunction may play an important role of a certain pathogenesis of GN.

Acute vascular rejection (AVR) in kidney transplantation is the most important factor influencing graft prognosis. We focus on patients whose grafts were lost because of AVR, and assessed their clinical characteristics and histological findings of biopsied renal grafts. Biopsied specimens exhibited AVR in 43 patients who underwent kidney transplantation in the Kidney Center of Tokyo Women's Medical University from 1995 to 1999. In the follow-up from 1 to 5 yr (median: 2.5 yr) we classified these patients into three groups: favourable prognosis group (FPG), relatively poor prognosis group (RPPG) and poor prognosis group (PPG). Light microscopic study for histological grading of acute rejection according to the Banff scheme and detection of the C4d complement deposition on peritubular capillaries by the immunofluorescence method were performed. Based on the results, the donors of RPPG and PPG were significantly older than those of FPG, and all factors of acute rejection according to the Banff scheme were not statistically significantly different among the three groups. However, an acute tubular injury mimicking acute tubular necrosis (ATN) was observed in the biopsy specimens from PPG. In conclusion, an older donor is a risk factor of poor prognosis of the graft with AVR, and acute tubular injury mimicking ATN is one of the important features that enables the prediction of graft failure originating from AVR in kidney transplantation.

To elucidate the role of osteopontin (OPN) in monocyte recruitment in crescentic glomerulonephritis, we investigated immunohistochemical localization of OPN in the kidney and its correlation with clinical and histopathologic parameters in biopsy specimens of patients with myeloperoxidase antineutrophil cytoplasmic autoantibody- (MPO-ANCA) associated glomerulonephritis.Twelve patients with MPO-ANCA-associated glomerulonephritis were enrolled in this study. Clinical parameters such as creatinine clearance and urinary protein excretion of each patient were obtained at the time of biopsy. Paraffin-embedded sections were used for immunohistochemical staining using the LSAB method. Five cortical interstitial fields randomly selected at original magnification x 200 were assessed using a computer-assisted color image analyzer. Tubular OPN expression was assessed as the percentage of positive area in the tubulointerstitium. Double immunofluorescent staining using antibodies against OPN and alpha(v)beta3 was performed.In all of the cases studied, OPN was occasionally localized within the glomeruli, and expressed slightly in proximal tubular epithelium and significantly in distal tubular epithelium. Tubular OPN expression tended to be promoted in the interstitium infiltrating by numerous monocytes/macrophages. The extent of tubular OPN expression was positively correlated with serum ANCA titers and urinary OPN concentrations. Enhanced alpha(v)beta3 expression appeared in the distal tubular epithelium expressing OPN.These results suggest that inducible expression of OPN and alpha(v)beta3 in the tubular epithelium seems to be associated with interstitial moncyte infiltration and subsequent tubulointerstitial changes in human MPO-ANCA-associated glomerulonephritis.

Post-translational modifications play diverse biological functions. Hydroxylation of collagen proteins has long been a recognised post-translational modification in eukaryotes. In the case of collagen, hydroxylation of prolyl residues, by 2-oxoglutarate and iron dependent enzymes (2OG oxygenases), in collagen proteins allows for the stabilisation of the collagen triple helix structure through conformational restraint and through the addition of a hydrogen bond donor. Additionally, hydroxylation of lysine side chains of collagen is required for cross-linking collagen (and possibly other proteins) in the extra-cellular matrix. Post-translational prolyl hydroxylation also plays a pivotal role in transcriptional regulation of the hypoxic response, as catalyzed by the hypoxia inducible factor / HIF prolyl hydroxylases (PHDs or EGLN enzymes). Recently, ribosomal protein hydroxylation catalyzed by 2OG- and Fe(II)-dependent oxygenases has been found to be a highly conserved post-translational modification in eukaryotes and prokaryotes (Ge et al and Loenarz et al). We present several crystal structures of 2OG oxygenases involved in ribosomal protein hydroxylation.

Tacrolimus (TAC) is a useful immunosuppressive agent in the prevention of rejection. However, the blood level between its therapeutic and toxic levels is narrow such that its nephrotoxicity is a problem. Moreover, its bioavailability and pharmacokinetics are highly variable. We experienced a case of acute nephrotoxicity, in which the blood level rose about 10 times above the expected level. We found a peculiar vacuolization in the transplant biopsy specimen. This change showed a marked vacuolization of the tubular cells, suggestive of acute nephrotoxicity by TAC.

Two long-term renal transplant survivors, for about 20 yr, with unusual histological features in the allograft kidney are reported. In both cases, marked hyalinosis was observed in the arterioles of the transplanted kidney, despite never having been administered cyclosporine or tacrolimus. The cause remains unknown at the present time, but we think that the changes could be multifactorial in origin, including due to aging of the graft, hypertension, hyperlipidemia and chronic rejection. We conclude that histological analysis of the allograft kidney must be performed in long-term renal transplant survivors, in order to understand the histological changes in the chronic phase after kidney transplantation and to predict the prognosis of the graft.

To investigate the serum levels of vascular endothelial growth factor (VEGF) in human crescentic glomerulonephritis (CRGN).The serum VEGF levels in CRGN patients were compared with those in healthy subjects and in various types of glomerulonephritis. In addition, we investigated the relationship between serum VEGF levels and creatinine levels (Scr) and pathological parameters.The serum VEGF levels of the CRGN patients were significantly higher than in the healthy subjects and patients with MCNS, IgAN, and FGS. No correlation was found between serum VEGF levels and Scr in the RPGN patients. The serum VEGF levels in 6 CRGN patients had significantly decreased after 6 months of corticosteroid therapy. Moreover, there was a significant correlation between the serum VEGF levels and the crescent frequency or the grade of interstitial injury, and the rates of glomerular infiltration by macrophages.In the CRGN patients, severe glomerular and interstitial damages would result in increased serum VEGF levels and corticosteroid therapy may exert its efficacy through reduction of serum VEGF levels.

The present study was designed to elucidate whether glomerular epithelial cell (GEC) injury is associated with the mechanism of proteinuria in biopsy-proven IgA nephropathy (IgAN). Twenty-four IgAN patients were divided into 4 groups based on their grade of proteinuria. Light microscopic examination revealed that GEC injury appeared prior to glomerular lesions such as sclerosis, crescent and adhesion. Reduced glomerular expression of C3b receptor (CR1), an indicator of GEC injury, was detected initially at the portion of damaged GEC and around the lesions of sclerosis, crescent and adhesion. CR1 expression eventually disappeared in the group IV patients who had nephrotic range proteinuria. Moreover, the grade of proteinuria in IgAN patients was associated with GEC injury and reduced glomerular expression of CR1. Taken together, GEC injury might be an important pathological finding which appears initially in the glomeruli, suggesting that GEC injury is a predictor of proteinuria in IgAN patients.

There is a strong association between epithelial crescent formation and the outcome of patients with Henoch-Schönlein purpura nephritis (HSPN), but little is known about the mechanisms of crescent formation in this disease. In this study, we examined whether or not glomerular endocapillary proliferation, fibrinoid necrosis and infiltrating inflammatory cells are involved in the process of crescent formation. Renal biopsy specimens were obtained from 19 patients (11 males and 8 females, 7.8 +/- 3.8 years old). The biopsies were performed within the initial 3 months after the onset of urinary abnormalities in all patients. Twelve patients had crescents (% glomeruli with crescents; 27.5 +/- 20.4%), and the other 7 did not. Renal tissues biopsied from 10 patients with asymptomatic hematuria served as controls for quantitative studies of glomerular leukocyte infiltration. Prominent endocapillary proliferation as well as fibrinoid necrosis were encountered in patients with crescents in contrast to patients without crescents. A positive correlation between crescents and endocapillary proliferation or fibrinoid necrosis was also observed. Significant increases in the number of glomerular CD68+ macrophages, CD45RO+ T cells and NP57+ neutrophils were observed in HSPN patients compared with the controls. When patients with or without crescents were compared, patients with crescents had a greater number of inflammatory cells. Although all kinds of inflammatory cells appeared in the endocapillary proliferative lesion, only CD68+ macrophages and NP57+ neutrophils were present in the fibrinoid necrotizing lesion. In conclusion, the present study demonstrated that necrotizing lesions with capillary rupture may be essential in the process of crescent formation seen in HSPN. Although the mechanism by which capillary wall breaks occur is still unknown, this study indicates that infiltrating macrophages as well as neutrophils may be involved in capillary rupture leading to epithelial crescent formation.

Contraction of the pre- and postglomerular arteries play an important role in the regulation of glomerular blood flow. Mesangial cells may also be involved in the mechanism of this regulation, but it has not been clarified yet whether or not mesangial cells and vascular smooth muscles show an identical phenotype, especially in terms of their contractile proteins. In this study, in order to elucidate any difference in the cellular phenotypes between mesangial cells and renal vascular smooth muscles, we investigated the localization of myosin heavy chain isoforms using a monoclonal antibody against SM1 and SM2. Both SM1 and SM2 are specific to smooth muscles. SM1 is specifically expressed in smooth muscles from early development and SM2 appears after birth. In normal renal tissues, SM1 and SM2 were expressed only in the smooth muscle cells of the arterioles and small arteries. However, glomerular cells, including mesangial cells, were not stained with either anti-SM1 antibody or anti-SM2 antibody. Localization of SM1 and SM-2 was similar to that of alpha-smooth muscle actin (SM alpha-actin). Staining for SM-1 was not observed in the mesangial areas of renal tissues with glomerular disease. These results clearly indicate that mesangial cells have a different phenotype from that of vascular smooth muscle cells in terms of their contractile proteins.

To investigate the mechanism of glomerular endothelial response to high glucose, an in vitro model of the glomerular endothelial barrier was established in which transfer of fluorescein-labeled albumin across confluent monolayers of immortalized bovine glomerular endothelial cells (GEN) grown on polycarbonate membranes was measured. We first examined the effects of increased concentrations of D-glucose on albumin permeability across GEN monolayers and further investigated the role of protein kinase C (PKC) in the regulation of glucose-induced changes in endothelial barrier function. Incubation with 30 mM D-glucose increased albumin permeability more than those with 10 mM D-glucose. Albumin permeability incubated with 10 mM D-glucose plus 20 mM mannitol was not significantly different from those with 10 mM D-glucose, indicating that the increase in albumin permeability induced by 30 mM D-glucose was not due to high osmolar stimuli. A protein kinase C (PKC) inhibitor, H-7 (25 microM) significantly reduced the permeability-increasing effects of D-glucose. Lactate dehydrogenase release from endothelial cells was not significantly increased above baseline after incubation with 10 mM or 30 mM D-glucose. These results suggest that elevated concentrations of glucose activates PKC, resulting in an increase in albumin permeability across GEN.

Infiltration of leukocytes and mononuclear cells into the glomeruli is an early pathological finding in human and experimental glomerulonephritis. However, the cellular and molecular basis for cell infiltration into the glomeruli is not yet completely understood. In addition, there is little information on the expression of intercellular adhesion molecule-1 (ICAM-1) on glomerular cells. In the present study, we investigated the expression of ICAM-1 on cultured bovine glomerular endothelial cells (GEN) and its regulation by the pro-inflammatory cytokines, interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1) and lipopolysaccharide (LPS). Immunocytochemical staining showed that ICAM-1 molecules were constitutively expressed on the surface of GEN. In flow cytometric and ELISA analyses, ICAM-1 molecule expression on GEN was significantly upregulated by IL-1 beta, MCP-1 and LPS in a dose-dependent manner, but not by IL-6. LPS was the most potent inducer of ICAM-1 molecule expression on GEN. The effects of IL-1 beta, MCP-1 and LPS were observed as early as 4 h and reached a maximal level by 18 h. These results suggest that ICAM-1 on GEN can participate in the infiltration of mononuclear cells into glomeruli in human and experimental glomerulonephritis.

The importance of unravel hemodynamics at the stomach and esophagus in patients with portal hypertension has been described for understanding pathophysiology and treatment of varices associated with portal hypertension. Microvascular Doppler sonogrlaphy (MF20) is a pulse Doppler method with 20 MHz which is able to measure velocity and direction of blood flow in microvessel. We invented transendoscopic microvascular Doppler sonography (EMDS) by connecting a tiny probe of one mm in diameter with an endoscopy. We evaluated direction of blood flow in the esophageal and gastric varices by EMDS and angiography in 21 untreated and 26 shunt-operated patients with portal hypertension. In all of untreated patients direction of blood flow in the varices was toward the probe (forward). Results obtained with EMDS were consistent with angiographic results available in 13 patients. Direction of blood flow after shunt operation was away from the probe in 21 out of 26 patients. The comparison with angiography revealed that post-operative patients with "away" blood flow in varices had maintained satisfactory shunt selectivity without hepatofugal collateral and it proved that the direction of blood flow was toward the shunt at lower pressure. In conclusion EMDS is useful as an evaluation method for hemodynamics in varices of the esophagus and stomach.

To examine the indication of nephron sparing operations, we searched 31 nephrectomized kidneys with renal cell carcinoma for small associated lesions in the normal-appearing portion of these kidneys. The diameter of the predominant tumors was between 15 and 50 mm. The kidneys were serially sectioned at 5-mm intervals, and all sections were examined microscopically. Two of the 31 kidneys had a small renal cell carcinoma, and four had a small adenoma in the normal-appearing portion. We recommend that the nephron-sparing operation should be is performed limitedly and that radical nephrectomy is yet a standard operation in the presence of a normal contralateral kidney.

Several clinical trials have demonstrated that cisplatin-based chemotherapy for primary intracranial germ-cell tumors is effective as a neoadjuvant chemotherapy. In this report, we describe a 6-year-old boy, Down syndrome and Hirschsprung's disease with intracranial pure yolk sac tumor treated by combined chemotherapy with cisplatin, vinblastine, bleomycin and cyclophosphamide (modified VAB-6 regimen). He had been admitted to our hospital because of intractable vomiting, and left facial nerve palsy since 1 month before. An MRI revealed an enlarged mass, 4cm in diameter, in the left cerebello-pontine angle with uniformal enhancement by Gd-DTPA, and bilateral ventricular dilatation. He was found to have increased serum alpha-fetoprotein level (AFP 11, 786ng/ml), but not human chorionic gonadotropin beta-subunit. After a partial resection of the tumor, diagnosed as pure yolk sac tumor, and ventriculo-peritoneal shunt, three courses of combined chemotherapy with cisplatin, bleomycin, vinblastine and cyclophosphamide (modified VAB-6 therapy) were carried out. The serum AFP level returned to normal, and the tumor mass entirely disappeared (a complete response) on MRI after the second course of chemotherapy. However, cisplatin-induced vomiting and mild neutropenia and renal tubular injury developed after the third course of chemotherapy. Irrespective of administration of recombinant human G-CSF and broad spectrum antibiotics, he suffered from pneumonia and died of septic shock and multiple organ failure. Autopsy showed microscopic residual tumors. The combination chemotherapy with cisplatin, bleomycin, vinblastine and cyclophosphamide is effective for initial treatment of childhood intracranial yolk sac tumor. It is necessary, however, to reevaluate the cisplatin dosage and treatment schedule in order to reduce such side effects as bone marrow suppression and renal damage.

A 49-year-old woman suffering from nephrotic syndrome (NS) was admitted for renal biopsy and treatment of NS. Light microscopy demonstrated that the glomerular capillary wall was markedly thickened with diffuse accumulation of periodic acid Schiff- and periodic acid methenamine-positive materials, leading to lobular accentuation of glomerular tufts. By electron microscopy, numerous collagenous fibers were observed in the mesangium and subenodothelial area. The fibers were peculiarly curved and frayed, as reported in nail-patella syndrome. These materials were thought to be type I and type III collagen as a result of immunohistochemical studies. No laboratory data or pathological findings were found to be compatible with previously described glomerulonephritis. The primary glomerular fibrosis in the present patient seemed to be a case of collagenofibrotic glomerulonephropathy.

We conducted a study to investigate whether expression of the contractile proteins in cultured rat mesangial cells (MC) was associated with the cell cycle and protein kinase C (PKC). When growth-arrested MC were stimulated with 100 nM phorbol myristate acetate (PMA) for 24 hours, an increased expression of smooth muscle alpha-actin and vimentin was detected by immunocytochemistry. A proportion of the S- and G2/M-phase in MC was increased in accordance with the enhanced expression of contractile proteins on flow cytometry. Immunoblot analysis revealed that 100 nM PMA stimulated expression of alpha-actin and vimentin as a single band. These results indicate that expression of contractile proteins, such as alpha-actin and vimentin, is dependent on the cell cycle and PKC, suggesting a phenotypic change in which MC assume smooth muscle cell characteristics.

Helicobacter pylori has been shown to infect the gastric mucous layer of almost all patients with duodenal ulcer disease, as well as that of most patients with gastric ulcer disease. Recent studies have suggested that the eradication of H. pylori affects the natural history of duodenal ulcer disease such that the rate of relapse decreases markedly. We evaluated the relationship between H. pylori infection and peptic ulcer relapse in a Japanese population following a prospective study. Seven of 18 (38.9%) gastric ulcer patients positive for H. pylori relapsed by the end of 1 year, whereas only 1 of 9 (11.1%) gastric ulcer patients without H. pylori developed ulcer relapse (P0.05). Relapse rates of duodenal ulcer patients negative for H. pylori were significantly lower than those positive for H. pylori within 1 year (0% vs 66.7%; P0.01). The effects of anti-H. pylori drugs on the eradication of H. pylori were examined in 50 patients with peptic ulcers. Eradication rates with a proton pump inhibitor (PPI) alone (omeprazole 20 mg) showed the lowest values (4 of 13; 30.8%). The rates were: 44.4% for amoxicillin alone (4 of 9); 70% for triple therapy consisting of amoxicillin, metronidazole, and bismuth subnitrate (14 of 20); and 87.5% for concomitant therapy of the PPI plus amoxicillin (7 of 8). Reinfection rates of H. pylori within 1 year after eradication of this organism were distinctly higher in the PPI alone group (80%) than in other groups (18.2%-32.4%).(ABSTRACT TRUNCATED AT 250 WORDS)

In this study, we have investigated the clinical effectiveness of fluconazole (FLCZ) given intravenously or orally to pediatric patients with systemic fungal infections. FLCZ was administered intravenously to two patients with acute leukemia (multiple hepatosplenic candidiasis and aspergillosis) and orally to two mycosis complicated with neuroblastoma and aplastic anemia, respectively. Clinical efficacies were excellent and no side effects were observed in any patients. Pharmacokinetic analysis in 6 neonates revealed that the plasma half-life is 37-41 hours after administration of single dose of intravenous infusion of 3 mg/kg of FLCZ.

A new monoclonal antibody (MAb) was raised against human glomerular components to investigate the possible existence of glomerular antigens reflecting cellular adaptation in glomerulonephritis. The MAb recognized a podocyte antigen as well as those expressed on renal arterial endothelial cells and smooth muscle cells by indirect immunofluorescence. An additional, but weaker immunoreaction was found in epithelial cells of the Bowman's capsule. This MAb recognized a 30-kD protein on western blotting of glomerular extracts under non-reducing and reducing conditions. Immunoperoxidase electron microscopy revealed that this antigen is present within the cytoplasm, but not on the cell membrane of the podocytes. Moreover, the antigen was found to be reduced in the glomeruli of patients with minimal change glomerulonephritis. These results suggest that the 30-kD protein is a novel protein, which we hypothesize is involved in maintenance of the structural and functional integrity of the podocytes. In addition, reduced expression of the 30-kD protein in the podocytes may be related to the increasing proteinuria in minimal change glomerulonephritis.

We report a hemodialysis patient with an atypical renal neoplasm. The tumor cells were arranged in a two-cell pattern similar to that in the usual excretory duct systems. The histochemical staining pattern with some lectins and monoclonal antibody corresponded to the distal nephrons of normal kidney tissue. These findings enabled us to diagnose this patient as having so-called Bellini's duct tumor.

The chronic administration of cyclosporin A (CyA) could be associated with renal arteriolar damage. Endothelial cell injury is considered to be the first event in this pathological change. Bovine glomerular endothelial cells (GEN) were used to investigate whether CyA induces microvascular injury in vitro. The damage induced by CyA was demonstrated by cell detachment from the culture plate and cell lysis as characterized by the increase in lactate dehydrogenase (LDH) at the same time after CyA treatment. One mumol/L of CyA induced detachment of GEN within 3 h and lysis after 6 h. Cyclosporin A-induced GEN lysis appeared in a dose and time-dependent manner. These results indicate that GEN are vulnerable to CyA exposure, suggesting that GEN damage might be related to CyA-mediated glomerular injury.

In this study, we measured cMLC1 concentration in serum and urine from patients with acute myocardial infarction (AMI), chronic renal failure (CRF), and various grades of renal dysfunction (RD) in comparison with normal controls, by using enzyme immunoassay (EIA) with monoclonal antibody, and attempted to elucidate the mechanism of increased serum level of cMLC1 in patients with renal failure. The serum level of cMLC1 of CRF patients under maintenance hemodialysis (HD) was 20.3 +/- 19.6 ng/ml, markedly higher than normal controls (0.54 +/- 0.55 ng/ml). The patients with RD and CRF under conservative therapy had higher serum cMLC1 level than normal controls especially in advanced CRF, while each value not correlating with their creatine clearance (Ccr). cMLC1 in urine was detectable in only two cases with AMI accompanied with CRF or RD. In addition, immunohistological studies of renal biopsy specimens from RD patients did not show cMLC1 deposits in glomerulus. These results suggest that cMLC1 is assumably filtered through the glomerulus, and then absorbed in the renal tubule.

The effect of recombinant human interleukin 6 (rhIL-6) on the in vitro growth of human bone marrow myeloid progenitors (granulocyte-macrophage colony-forming units, CFU-GM) was investigated. Recombinant human IL-6 by itself did not induce colony formation. When rhIL-6 at various concentrations was added to the CFU-GM colony cultures containing recombinant human granulocyte colony-stimulating factor (rhG-CSF) or recombinant human granulocyte-monocyte/macrophage colony-stimulating factor (rhGM-CSF), rhIL-6 significantly suppressed the colony formation induced by rhG-CSF, but not by rhGM-CSF. This suppressive effect of rhIL-6 on rhG-CSF-induced, but not rhGM-CSF-induced colony formation was confirmed by using an MY10(+)-cell-enriched population. Neither interleukin 3 nor interleukin 1 alpha suppressed the growth of myeloid progenitors. The preincubation of bone marrow cells with rhIL-6 for a short time (30 min) resulted in a reduction of colonies induced by rhG-CSF, but not by rhGM-CSF. The suppressive effect of rhIL-6 on rhG-CSF-induced colony formation was not observed when the cells were preincubated together with rhG-CSF at a high ratio of rhG-CSF to rhIL-6. The rhIL-6-mediated suppressive effect was further confirmed by blocking the effect by the anti-IL-6 antibody. These results suggest antagonistic interaction between IL-6 and G-CSF in the later differentiation of myeloid progenitors.

Gel shift assays have been employed to examine the association of the thyroid hormone receptor with specific DNA sequences in the 5′-flanking DNA of the rat growth hormone (rGH) gene. This DNA is known to have structure(s) that mediate thyroid hormone effects on the rGH promoter. The receptors used were obtained from preparations purified 300-500-fold from rat liver nuclear extracts and contained about 1% pure receptors. Thyroid hormone receptor binding to DNA was assessed by monitoring protein-bound 32P-labeled restriction endonuclease fragments in parallel with L-tri[125I]iodothyronine-labeled protein-DNA complexes. The receptors were found to bind specifically to four different regions of the rGH 5′-flanking DNA (nucleotides -1730 to -1230, -530 to -230, -181 to -149, and -149 to +12) numbered with respect to the transcriptional start site. The specificity of the binding was documented by the finding that the receptor did not bind to other rGH 5′-flanking DNA sequences or to several other DNAs and by the fact that only the DNAs exhibiting specific binding could block the binding of radiolabeled DNA. The binding was also detected in NaCl concentrations up to 140 mM, reduced by Mg2+ concentrations up to 5 mM, and inhibited by 1 mM zinc. The DNA sequence-specific binding of the receptor was found to require occupancy of the receptor by the hormone (L-triiodothyronine) and could also be observed when the receptor was occupied by the thyroid hormone antagonist amiodarone. These results indicate that thyroid hormone receptors interact specifically with several sites on the 5′-flanking DNA of the rGH gene and that hormone occupancy is not required for the binding. Thus, thyroid hormone may act by stimulating a transcriptional activation function of the receptor rather than by stimulating DNA binding per se.

Phosphorylation of tyrosine residue (Y1204) of rat nephrin by Fyn kinase allows Nck adaptor protein binding to nephrin motifs, which include the phosphorylated tyrosine. This phosphorylation-dependent switch induces actin polymerization in a cell culture system. Here, we generated an antibody recognizing phosphorylated nephrin at the Nck binding sites pY1204 and pY1228 to determine the phosphorylation status of nephrin using a rat model of puromycin aminonucleoside-induced nephrosis. Changes in globular actin (G-actin) and filamentous actin (F-actin) contents in isolated glomeruli were measured by western blot. Before experimental nephrosis, both Y1204 and Y1228 were phosphorylated, and most of the actin was filamentous. Before the onset of overt proteinuria, however, phosphorylation of both Y1204 and Y1228 rapidly decreased and became almost undetectable. During this period, the amount of F-actin in glomeruli began to decrease, whereas G-actin increased. Phosphorylation of nephrin at Y1228 in glomeruli of patients with minimal change nephrosis was significantly decreased compared with that in normal glomeruli. Our study suggests that tyrosine phosphorylation of nephrin by regulating F-actin formation may be important for the maintenance of normal podocyte morphology and function.

Ia-like antigen-positive (Ia+) and -negative (Ia-) cell populations were separated from human cord blood cells and bone marrow mononuclear cells by a rosette technique with a combined use of staphylococcal protein-A-coated bovine red blood cells and the monoclonal OKIa 1 antibody, or by using a cell-sorting technique. Colony-forming units-granulocytes-monocytes-macrophages (GFU-GM) were assayed in a semisolid agar culture, and colony-forming cells-granulocytes (CFC-G) were differentiated from colony-forming cells-monocytes-macrophages (CFC-M) by double staining for esterase activity. The majority of CFC-G in cord blood was grown in the Ia+ fraction; Ia+ CFC-G/Ia- CFC-G = 1.62 +/- 0.34 (mean +/- SD), which was similar to the ratio in bone marrow (Ia+/Ia- = 1.80 +/- 0.37). In contrast, the majority of CFC-M in cord blood was grown in the Ia- fraction; Ia+/Ia- for CFC-M = 0.50 +/- 0.09. The predominance of CFC-G in the Ia+ fraction in contrast to predominance of CFC-M in the Ia- fraction was confirmed by using a cell-sorting technique. T-lymphocyte depletion and the culture supernatants of Ia+ and Ia- cells did not affect differentiation of CFC-G and CFC-M. These data suggest that there are potent differences in the expression of Ia-like antigens between CFC-G and CFC-M, indicating that the Ia+ progenitor cell population generates predominantly CFC-G, whereas the Ia- population generates mainly CFC-M during the maturation process in granulopoiesis.