Focal segmental glomerulosclerosis and partial deletion of chromosome 6p: a case report.

We treated a patient with 6p partial deletion syndrome diagnosed after proteinuria was detected during developmental examination 3 years after birth. External anomalies included ocular hypertelorism, saddle nose, elongated philtrum, tent-like lips, and low-set auricles. Mental retardation was evident. The karyotype was 46,XX,del(6) (p.22.1-p22.3) with an interstitial deletion. The kidneys showed no abnormality on imaging such as hydronephrosis, atrophy, or malformation. Examination of a renal biopsy specimen disclosed focal segmental glomerulosclerosis. No cardiac anomaly or Rieger anomaly, which often are present in this syndrome, were noted.

An adolescent with marked hyperimmuno-globulinemia E showing minimal change nephrotic syndrome and a STAT3 gene mutation.

We encountered a patient with marked hyperimmunoglobulinemia E who had a mutation of the signal transducer and activator of transcription 3 gene (STAT3) and developed minimal change nephrotic syndrome (MCNS). From early infancy, the patient showed repeated episodes of refractory chronic eczema accompanied by impetigo vulgaris with cicatrization, as well as otitis media. Serum IgE was markedly increased (from 4,000 to 25,000 IU/ml). The nephrotic syndrome (NS) frequently relapsed, and was alternately responsive and resistant to corticosteroids. The STAT3 mutation was heterozygous, located in exon 23 of the transactivation domain and causing A744V substitution. Presently treated with mycophenolate mofetil, the patient has less frequent MCNS recurrences. Increases in circulating Th2 cytokines and IgE combined with suppression of the Th1 cytokine interferon-γ caused by the STAT3 abnormality, presumably caused MCNS by altering the Th1/Th2 balance among T-lymphocytes. To our knowledge, this is the first report of type I hyper-IgE syndrome (HIES) showing a STAT3 gene mutation and MCNS.

A female infant with Frasier syndrome showing splice site mutation in Wilms' tumor gene (WT1) intron 9.

Wilms' tumor gene (WT1) abnormality leads to various disorders of differentiation as well as renal and urinary system abnormalities. Here we present a case of WT1 abnormality and steroid-resistant nephrotic syndrome in a female infant. The 3-year-old patient was initially diagnosed with proteinuria at an annual mass screening program for children aged three years and was referred to our hospital. She met the diagnostic criteria for nephrotic syndrome and showed normal renal function. The patient was treated with corticosteroids; however her condition showed resistance to corticosteroids. On renal biopsy, she was diagnosed with focal segmental glomerulosclerosis (FSGS). Because of the possibility of WT1 abnormality, an exon array analysis was conducted, which ruled out Denys-Drash Syndrome (DDS). The patient was then diagnosed with Frasier Syndrome (FS) on the basis of donor site mutation (IVS9+5G > A) of the splice site in the intron 9. Reports of female infants with FS are extremely rare. FS is one of the pre-mRNA splicing diseases, in which the occurrence of symptoms is associated with a decrease in the ratio of the lysine-threonine-serine (+/- KTS) isoform of the WT1 protein. A typical case exhibits 46 XY male karyotype and is characterized by male pseudohermaphroditism with cord-like gonadal structures as well as progressive nephropathy caused by FSGS. However, in female infants without such extrarenal signs, it is necessary to consider the analysis for WT1 intron 9 for conclusive diagnosis of FS, because the presence of nephropathy is the only symptom for possible detection.

A case of renal-coloboma syndrome associated with mental developmental delay exhibiting a novel PAX2 gene mutation.

A case of an adolescent male with renal-coloboma syndrome (RCS) showing developmental delay is described. Birth and perinatal histories were typical. Proteinuria was initially observed at the age of 7 years during an annual mass screening program for school children. His urine was checked periodically at a local hospital. Because of an increase in proteinuria, he was referred to our hospital for further clinical evaluation. Proteinuria was moderate, ranging from 1.0 to 1.5 g/day, and was coupled with mild renal dysfunction. At that time, he was found to have myopia associated with astigmatism. He exhibited mild developmental delay, assessed by a WISC-III test. A renal biopsy sample showed marked glomerular enlargement, collapse of glomerular capillaries, mesangial matrix expansion, and tubulointerstitial change, demonstrating typical histologic features of RCS. Approximately five years after starting follow-up, the patient had severe renal dysfunction. Furthermore, optic nerve coloboma was also evident. Genetic analysis of the patient revealed a novel heterozygous mutation in exon 3 of the PAX2 gene (P130H).

Association of INVS (NPHP2) mutation in an adolescent exhibiting nephronophthisis (NPH) and complete situs inversus.

Cystic kidney disease has been linked to mutations in the Invs gene in mice with an inversion of embryonic turning (inv/inv) and the INVS (NPHP2) gene in human infantile nephronophthisis (NPH). Infantile NPH shows marked cyst formation in contrast to other forms of NPH and rapidly progresses to end-stage renal failure (ESRD) before 5 years of age. In this report, we describe an adolescent with a mutation in INVS who had preservation of his renal function beyond infancy. The patient showed findings of NPH with mild renal insufficiency together with situs inversus. He also exhibited a series of features consistent with Jeune syndrome involving asphyxiating thoracic dystrophy, heart failure and hypertension prior to advanced renal insufficiency. Based upon these features, our patient is likely to have the combined clinical features of infantile NPH with Jeune syndrome. Genetic analysis for INVS disclosed a heterozygous mutation of TrG at position rs7024375 in the 5'UTR of INVS in the patient and his mother, while no abnormalities were found in any of the 17 exons of INVS or NPHP1, 3 and 4. To our knowledge, this is the first patient possessing a genetic alteration in INVS who had preservation of renal function past childhood. This study suggests that our patient may be a compound heterozygote for infantile NPH and Jeune syndrome, because both these disorders are transmitted mainly as an autosomal-recessive trait.

Chronic renal insufficiency in a boy with cystic renal lymphangiectasia: morphological findings and long-term follow-up.

Cystic renal lymphangiectasia (CRL) is a rare malformation of lymphatics that can present in childhood and adulthood. Symptoms and radiologic features are relatively well defined, but clinical evolution and prognosis remain unclear. We treated a boy with CRL who developed chronic renal insufficiency. The first manifestation was abdominal swelling associated with an umbilical hernia noted incidentally at 1.6 years. Computed tomography with intravenous contrast administration demonstrated perirenal cysts with fluid collection, suggesting CRL. Intractable ascites resisted pharmacologic treatments such as diuretics. After approximately 7 years, the ascites resolved spontaneously, but the perirenal cysts persisted. At 11 years, proteinuria was noted. A renal biopsy specimen showed interstitial abnormalities consistent with CRL, glomeruli showed a focal segmental mesangial increase. Proteinuria persisted despite administration of an angiotensin-converting enzyme inhibitor, increasing as obesity and hypertension worsened. Renal function gradually declined in the ensuing years. Polycythemia coexisted with a normal serum erythropoietin concentration. A follow-up renal biopsy specimen disclosed glomerular enlargement together with focal segmental mesangial expansion, suggesting obesity-related glomerulopathy. Our observation suggest that under some specific circumstances like our patient CRL may exacerbate. Management of complicating obesity and hypertension are likely to be important for maintaining normal renal function, especially in the diffuse bilateral type of CRL present in our patient.

A Japanese family with Alport syndrome associated with esophageal leiomyomatosis: genetic analysis of COL4A5 to COL4A6 and immunostaining for type IV collagen subtypes.

BACKGROUND: In some families, X-linked Alport syndrome (AS) is associated with diffuse leiomyomatosis. We describe clinical, pathologic and molecular-genetic findings in a Japanese family with this inheritance mode of AS in association with leiomyomatosis. PATIENT: AS was diagnosed in a one-year-old boy with recurrent aspiration pneumonia caused by esophageal stenosis from leiomyomatosis. Diagnosis was confirmed by electron microscopy coupled with type IV collagen chain subtype staining in a renal biopsy specimen. His mother, who exhibited esophageal leiomyomatosis and is heterozygous for AS, showed a discontinuous staining pattern for collagen alpha5(IV) chain along the epidermal basement membrane in a skin biopsy specimen. Genetic analysis in the boy revealed the deletion of the first two exons of COL4A6 together with deletion of the 5' end of COL4A5. Despite administration of cyclosporin A, massive proteinuria has persisted in the boy, although renal function otherwise remains normal. CONCLUSION: Identification of an AS patient during infancy is extremely rare. Clinical manifestations, including macroscopic hematuria, cataracts and leiomyomatosis caused by the large deletion involving COL4A5 to COL4A6, led to early presentation with AS.

A boy with Japanese Dent's disease exhibiting abnormal calcium metabolism and osseous disorder of the spine: defective megalin expression at the brushborder of renal proximal tubules.

We encountered a 16-year-old boy with Japanese Dent's disease who exhibited renal insufficiency and an osseous disorder of the spine. Proteinuria first was noted at the age of 2 years. At 13 years, the patient underwent analysis of the CLCN5 gene, which identified missense mutation (I524K) in exon 10. During follow-up, a marked increase in urinary beta2-microglobulin was associated with mild deterioration of renal function. At the age of 15 years, hypocalcemia (7.5 mg/dl) accompanied by an increased plasma concentration of alkaline phosphatase was first detected. At that time, plasma concentration of 25(OH)D3 and 1'alpha25(OH)2D3 were low accompanied by a high plasma parathyroid hormone concentration. A renal biopsy specimen revealed tubulointerstitial alterations including mononuclear cell infiltration, partial fibrosis and focal glomerular sclerosis. Immunofluorescence revealed weak, discontinuous staining of megalin along the brushborder of renal proximal tubules. Western blotting demonstrated decreased urinary excretion of megalin. Thus, clinical manifestations and prognosis may vary in Japanese Dent's disease. Reduced megalin expression may have disturbed calcium homeostasis, leading to osseous disorder in our patient.

Induction of inducible nitric oxide synthase in hepatocytes isolated from rats with ischemia-reperfusion injury.

BACKGROUND: Recent evidence indicates that nitric oxide (NO) has a crucial role in hepatic ischemia-reperfusion (I/R) injury. However, little is known about how I/R influences the gene expression of inducible nitric oxide synthase (iNOS) in hepatocytes. Under inflammatory conditions, we compared the induction of iNOS in hepatocytes isolated from normal and I/R-treated rats. METHODS: Hepatocytes were isolated using the collagenase perfusion method from rats treated with I/R (30-minute ischemia of middle and left lobes, followed by 3-hour reperfusion) or sham operation (control): Primary cultures of rat hepatocytes were incubated with an inflammatory cytokine, interleukin-1beta (IL-1beta), to compare the iNOS induction/NO production between the 2 groups. RESULTS: Both control and I/R groups had no production of nitrite (a stable metabolite of NO) in the absence of IL-1beta. In the control group, IL-1beta stimulated dose- and time-dependent production of NO. The I/R group showed more than 2-fold increased levels of NO production. Western and Northern blot analyses revealed that the I/R group also showed increased levels of iNOS protein and its messenger RNA. CONCLUSION: These results suggest that I/R directly affects the inducibility of the iNOS gene in hepatocytes by IL-1beta. Increased NO may be associated with protective or toxic effects in hepatic I/R injury.

Effects of pirfenidone on endotoxin-induced liver injury after partial hepatectomy in rats.

BACKGROUND: In living donor liver transplantation, restrictions on graft size are a serious obstacle to expand indications for adult recipients. The sequence of gram-negative infection, septicemia, and multiple-organ failure is a common cause of early mortality after liver transplantation. An effective therapy has not been established for endotoxemia following extended hepatectomy in donors or small-for-size grafts in recipients. Pirfenidone (PFD), a new experimental antifibrotic agent, was used to ameliorate on endotoxin-induced liver injury following partial hepatectomy. METHODS: Male Sprague-Dawley rats were intravenously administered lipopolysaccharide (LPS) 48 hours after 70% hepatectomy. Prior to LPS administration, PFD (300 mg/kg) or its vehicle (0.5% carboxymethylcellulose) was given orally twice. RESULTS: The survival rate of the PFD-treated group was markedly improved compared with that of the controls. PFD prevented the increases in the activities of serum enzymes (aspartate transaminase [AST], alanine transaminase [ALT], and lactate dehydrogenase [LDH]) and total bilirubin. The serum and liver tissue levels of inflammatory cytokines, such as tumor necrosis factor-alpha, interleukin-1beta, interferon-gamma, and interleukin-6, were significantly lower among the PFD than the control group. Furthermore, the degree of necrosis in the remnant liver was significantly decreased in the PFD-treated rats compared with controls. CONCLUSION: These results indicate that PFD alleviates endotoxin-induced liver injury after partial hepatectomy through the inhibition of production of inflammatory cytokines in the residual liver. PFD may be useful to prevent endotoxin-induced liver injury after hepatectomy.

Effect of hepatocyte growth factor on induction of cytokine-induced neutrophil chemoattractant in rat hepatocytes.

BACKGROUND: Restrictions on graft size are a serious obstacle to the expansion of indications for adult recipients in living donor liver transplantation. Hepatocyte growth factor (HGF) has a crucial role in regeneration following hepatic injury. Rat cytokine-induced neutrophil chemoattractant (CINC), a member of the interleukin-8 superfamily in humans, has been implicated in chronic liver diseases or development of liver ischemia-reperfusion injury. Studies were performed to examine whether HGF influences the induction of CINC in hepatocytes. METHODS: Primary cultures of rat hepatocytes were treated with or without recombinant human (rh) HGF. The release of CINC into the culture medium and levels of CINC mRNA were measured using an enzyme-linked immunosorbent assay and Northern blot analysis. Transcription of nuclear factor (NF)-kappa B was detected by electrophoretic mobility shift assays. RESULTS: rhHGF increased the release of CINC in the medium dose- and time-dependently, showing a maximal effect at 100 ng/mL. Genistein (100 mumol/L) blocked the release of CINC stimulated by rhHGF. Levels of CINC mRNA were also increased, reaching a maximum at 8 hours after addition of rhHGF. Electrophoretic mobility shift assays revealed rhHGF activated transcription factor, NF-kappa B. CONCLUSION: These results suggest that HGF stimulates the induction of CINC gene expression through activation of NF-kappa B. CINC may be involved in the function of HGF during liver regeneration.

Pirfenidone protects endotoxin-induced liver injury after hepatic ischemia in rats.

BACKGROUND: Pirfenidone (PFD), an experimental antifibrotic agent, was investigated for its effects on endotoxin-induced liver injury after hepatic ischemia-reperfusion. METHODS: Male Sprague-Dawley rats were subjected to 30 minutes of partial hepatic ischemia, followed by reperfusion for 24 hours. Lipopolysaccharide (LPS) was injected at 30 minutes of reperfusion. PFD (300 mg/kg) or its vehicle (0.5% carboxymethylcellulose) was given orally following LPS administration. RESULTS: PFD prevented the increase in activities of serum alanine transaminase, aspartate transaminase, and lactate dehydrogenase after reperfusion. PFD inhibited the increase of cytokine-induced neutrophil chemoattractant in serum and liver tissue. The number of neutrophils infiltrating the liver was significantly lower in the PFD-treated group than the control group. CONCLUSION: These results indicate that PFD prevents endotoxin-induced liver injury after hepatic ischemia-reperfusion, in part through the decrease of neutrophil infiltration to the liver.

Effect of pirfenidone on induction of chemokines in rat hepatocytes.

BACKGROUND: Hepatic ischemia-reperfusion results in a neutrophil-dependent liver injury. The process of neutrophil recruitment and activation in this injury is at least partially dependent on the induction of chemokines, such as cytokine-induced neutrophil chemoattractant (CINC) and macrophage inflammatory protein-2 (MIP-2) in rats. In the liver, parenchymal cells (hepatocytes), in addition to nonparenchymal cells such as Kupffer cells, have been reported to produce chemokines in the regulation of hepatic inflammation. Pirfenidone (PFD) is a new experimental drug used as an antifibrotic agent. Studies were performed to determine whether PFD influences the production of CINC and MIP-2 stimulated by interleukin (IL)-1beta in a primary culture model of rat hepatocytes. METHODS: Primary cultures of rat hepatocytes were treated with IL-1beta in the presence and absence of PFD. The protein and mRNA of CINC and MIP-2 were analyzed using enzyme-linked immunosorbent assays and Northern blots. RESULTS: IL-1beta increased the release of CINC and MIP-2 into culture media in a dose- and time-dependent manner. PFD inhibited both CINC and MIP-2 release in dose-dependent fashion. However, PFD had no effect on the levels of CINC mRNA induced by IL-1beta. CONCLUSION: These results suggest that PFD inhibits the production of CINC and MIP-2 by IL-1beta at a posttranscriptional step in hepatocytes.

Comparative strength of metal-ceramic and metal-composite bonds after extended thermocycling.

The relative strengths of ceramic-to-metal and composite-to-metal bonds were compared after prolonged thermocycling. A total of 104 cast discs were produced from a gold alloy (Pontor LFC). A ceramic material (Duceragold) was fused to 24 discs to assess the strength of the metal-ceramic bond. An indirect composite material (New Metacolor Infis) was bonded to the remaining discs after surface preparation by Rocatec tribochemical coating, tin plating and priming with a phosphate conditioner [10-methacryloyloxydecyl dihydrogen phosphate (MDP), Cesead II], priming with a thione conditioner (V-Primer) or no treatment (unprimed control). Shear bond strengths were determined before and after thermocycling at 20,000 and 100,000 cycles. Pre-thermocycling bond strengths were ranked in the order: metal-ceramic (40.5 MPa); Rocatec treatment (33.1 MPa) and tin plating-MDP (31.0 MPa); V-Primer (20.9 MPa); and control (11.9 MPa). The bond strengths of the first three groups were not significantly different after 20,000 thermocycles, whereas those of the V-Primer and control groups were significantly reduced. After extended thermocycling (100,000 cycles) the metal-ceramic group had the highest mean shear bond strength (28.5 MPa; P < 0.05), followed by the Rocatec (23.9 MPa) and tin plating-MDP (22.1 MPa) groups. The metal-ceramic bond was the most durable, although its strength was reduced by 29.6% after extended thermocycling. On the basis of these results, we recommend the Rocatec and tin plating-MDP systems for composite-to-metal bonding. Metal-ceramic bonding, however, is superior to metal-composite bonding within the limitation of the current experiment.

Alport syndrome and benign familial hematuria (thin basement membrane disease) in two brothers of a family with hematuria.

Alport syndrome (AS) and benign familial hematuria (BFH) are inherited disorders of the glomerular basement membrane, which are sometimes difficult to differentiate at the early stage without type IV collagen staining of the renal basement membrane. Previous studies have indicated that mutation of type IV collagen alpha4 gene may be responsible for both BFH and AS. We report here a Japanese family with consanguinity, in which autosomal-recessive AS and BFH were separately identified in two brothers on the basis of findings of electron microscopy and type IV collagen chain staining of the renal biopsy specimens. Their parents, being first cousins, paternal uncle and grandmothers were found to have hematuria. Our observations suggest that BFH patients were heterozygous carriers of autosomal-recessive AS.

Adhesive bonding of super-elastic titanium-nickel alloy castings with a phosphate metal conditioner and an acrylic adhesive.

The purpose of the current study was to evaluate the bonding characteristics of super-elastic titanium-nickel (Ti-Ni) alloy castings. Disk specimens were cast from a Ti-Ni alloy (Ti-50.85Ni mol%) using an arc centrifugal casting machine. High-purity titanium and nickel specimens were also prepared as experimental references. The specimens were air-abraded with alumina, and bonded with an adhesive resin (Super-Bond C & B). A metal conditioner containing a phosphate monomer (Cesead II Opaque Primer) was also used for priming the specimens. Post-thermocycling average bond strengths (MPa) of the primed groups were 41.5 for Ti-Ni, 30.4 for Ti and 19.5 for Ni, whereas those of the unprimed groups were 21.6 for Ti, 19.3 for Ti-Ni and 9.3 for Ni. Application of the phosphate conditioner elevated the bond strengths of all alloy/metals (P < 0.05). X-ray fluorescence analysis revealed that nickel was attached to the debonded resin surface of the resin-to-nickel bonded specimen, indicating that corrosion of high-purity nickel occurred at the resin-nickel interface. Durable bonding to super-elastic Ti-Ni alloy castings can be achieved with a combination of a phosphate metal conditioner and a tri-n-butylborane-initiated adhesive resin.

Chaotic mixing generated by acoustic streaming.

The acoustic mixer presented in this paper is a closed cylinder containing chemical solutions. The ultrasound was generated in a water bath outside of the cylinder. The mixing efficiency was measured by using a laser-photodiode system and by mixing iodine and sodium thiosulfate. Iodine solution has a light brown color which becomes transparent if the solution is mixed with sodium thiosulfate. The unmixed regions of the solution in the container remain dark. Starch was used to make the solution darker. Considering the relative position between the cylinder and the transducer, it was shown that displacements parallel to the axes of the streaming have little effect on the mixing speed. Indeed, the distance of the cylinder from the center of the streaming is more important to obtain ideal mixing performances. The frequency and size of the transducers has been investigated too and little effect was found. In contrast, the wave amplitude seems to be the biggest factor in obtaining rapid mixing.

Adhesive bonding of composite material to cast titanium with varying surface preparations.

The purpose of the present study was to evaluate the surface preparation effects of eight metal conditioners and an adhesive system on bonding between a prosthodontic composite material and cast titanium. Eight primers designed for conditioning base metal alloys (Acryl Bond, All-Bond 2 Primer B, Alloy Primer, Cesead II Opaque Primer, Eye Sight Opaque Primer, Metafast Bonding Liner, Metal Primer II, and MR Bond) as well as a surface modification technique (Siloc) were assessed. Disk specimens cast from titanium (T-Alloy H) were either primed with one of the eight primers or treated with the Siloc system, and then bonded with a light-activated composite material (Artglass). Bond durability was evaluated by thermocycling (4 and 60 degrees C, 1 min each, 20, 000 cycles). After thermocycling, two groups either primed with the Cesead II Opaque Primer material or treated with the Siloc system exhibited significantly greater bond strength (20.0 and 19.0 MPa) than the other groups (0.2-12.6 MPa, P < 0.05). These two systems are considered to be useful for improving bonding between the titanium and the composite material tested.

Shear bond strength of resin composite veneering material to gold alloy with varying metal surface preparations.

STATEMENT OF PROBLEM: Although adequate surface preparation is indispensable to achieve a consistent and durable bond between resin composite materials and the metal substructures of veneered restorations, information on the bonding performance of current metal adhesive systems is limited. PURPOSE: The purpose of this study was to evaluate the surface preparation effects of 4 metal conditioners and 1 adhesive system on bonding between a prosthetic resin composite veneering material and a gold casting alloy. MATERIAL AND METHODS: Four primers containing sulfur derivative monomer and designed for conditioning noble metal alloys (Alloy Primer, Infis Opaque Primer, Metal Primer II, and Metaltite) and a surface modification technique (Siloc) were assessed. Cast disk specimens made of gold alloy (Pontor LFC) were either primed with 1 of the 4 primers or treated with the Siloc system and bonded with a light-activated prosthetic resin composite material (New Metacolor Infis). Control specimens were also prepared without the use of a bonding agent. Shear bond strengths were determined before and after thermocycling (20,000 cycles) for evaluation of bond durability. RESULTS: All of the primed and Siloc-treated groups showed improved 24-hour shear bond strengths compared with the control group. After thermocycling, the groups either primed with the Metaltite conditioner or treated with the Siloc system exhibited the highest mean shear bond strengths. CONCLUSION: The Metaltite conditioner and Siloc system each represent a useful method for improving the bond between the gold alloy and resin composite material tested.