Review of renal tumors associated with Birt-Hogg-Dubé syndrome with focus on clinical and pathobiological aspects.

Birt-Hogg-Dubé syndrome (BHDS) is an autosomal dominant inherited disorder characterized by clinical features of skin lesions, pulmonary lesions and renal tumor. The gene responsible for this syndrome is located on chromosome 17p11.2 and designated as FLCN. In this article, we review renal tumors associated with BHDS with a focus on clinical and pathobiological aspects. Renal tumors often occur multifocally or bilaterally in the imaging analyses or gross examination. Histological examination of renal tumors includes a variety of subtypes such as hybrid oncocytic tumor, chromophobe renal cell carcinoma (RCC), oncocytoma, clear cell RCC and papillary RCC. The histologic discordance in multiple tumors seems to be characteristic of this syndrome. Oncocytosis is observed histologically in about half of the cases. Several investigations have elucidated that folliculin may be involved in the mammalian target of rapamycin (mTOR) pathway recently. Renal tumors composed of clear cells may behave in an aggressive fashion. However, renal tumors including hybrid oncocytic tumor, chromophobe RCC and oncocytoma behave mostly in an indolent fashion.

Correlation of autoantibodies and CD5+ B cells in ocular adnexal marginal zone B cell lymphomas.

AIM: To determine the clinicopathological properties of ocular adnexal marginal zone B cell lymphomas (MZBLs) with CD5+ B cells. METHODS: This study determined the clinicopathological properties of MZBL samples from 97 patients with ocular adnexal MZBLs and searched for hallmarks of systemic autoimmunity in these patients. RESULTS: Two elderly female patients were found to have ocular adnexal MZBLs with CD5+ B cells; flow cytometry analysis suggested that one of these MZBLs had CD5+ B cell clonal proliferation. The levels of anti-single stranded (SS)-DNA and anti-SS-A/Ro antibodies in these two patients were significantly higher than those in controls that were matched for age, gender and disease (2/2 versus 0/14; p = 0.008) and controls without MZBL (2/2 versus 0/30; p = 0.002). The genes from the immunoglobulin heavy-chain variable region for one of the patients showed a V3-21 segment. In addition, another patient with ocular adnexal reactive lymphoid hyperplasia with CD5+ B cells also had anti-SS-DNA antibodies. CONCLUSION: Patients with ocular adnexal MZBLs with CD5+ B cells may have a background of systemic conditions with CD5+ B-cell-related autoantibodies.

The effect of an associated ulnar styloid fracture on the outcome after fixation of a fracture of the distal radius.

A total of 118 consecutive patients with a fracture of the distal radius were treated with a volar locking plate; 50 patients had no ulnar styloid fracture, 41 had a basal ulnar styloid fracture, and 27 had a fracture of the tip of the ulnar styloid. There were no significant differences in radiological and clinical results among the three groups. The outcome was good and was independent of the presence of a fracture of the ulnar styloid. A total of five patients (4.2%) had persistent ulnar-sided wrist pain at final follow-up. Nonunion of the ulnar styloid fracture did not necessarily lead to ulnar wrist pain. Patients with persistent ulnar pain had a higher mean initial ulnar variance and increased post-operative loss of ulnar variance. The presence of an associated ulnar styloid fracture of the ulnar styloid does not adversely affect the outcome in patients with a fracture of the distal radius treated by volar plating.

Ovarian carcinoma recurring as carcinosarcoma.

Malignant mixed mesodermal tumor is a rare tumor of the ovary and its histogenesis is controversial. We report the case of an ovarian tumor that seemed to be a pure carcinoma and recurred as a carcinosarcoma, and suggest a possible histogenesis for this kind of tumor. The patient was a 62-year-old Japanese woman. The primary tumor was confined to the right ovary and was a histologically poorly differentiated endometrioid adenocarcinoma with focal squamous differentiation. The tumor recurred as peritoneal dissemination 9 months later showing a histological appearance of carcinosarcoma of heterologous type. The recurrent tumor also contained intermingled foci of similar histology as the primary tumor. The carcinomatous component of the recurrent tumor showed more obvious differentiation to adenocarcinoma with increased expression of epithelial markers compared to the primary tumor. Epithelial membrane antigen was positive also in a few cells of the sarcomatous component, which implies that this tumor had features of metaplastic carcinoma. The DNA ploidy pattern of the primary ovarian tumor was diploid, while an additional aneuploid subpopulation appeared in the recurrent tumor. These findings suggest the possible histogenesis of carcinosarcoma of the ovary as progression and clonal evolution of endometrioid adenocarcinoma.

P-glycoprotein-dependent disposition kinetics of tacrolimus: studies in mdr1a knockout mice.

PURPOSE: This study was performed to evaluate the involvement of P-glycoprotein in disposition kinetics of tacrolimus (FK506), a substrate of P-glycoprotein, in the body. METHODS: The blood and tissue concentrations of FK506 after i.v. or p.o. administration (2 mg/kg) to normal and mdr1a knockout mice were measured by competitive enzyme immunoassay. RESULTS: The blood concentrations in knockout mice were significantly higher than those in normal mice. The value of the total clearance (CLtot) for knockout mice (19.3 mL/min/kg) was about 1/3 of that for normal mice (55.8 mL/min/kg)(P < 0.001), although there was no significant difference in the distribution volume at the steady-state (Vd(ss)) (about 4.6 L/kg) between both types of mice. FK506 rapidly penetrated the blood-brain barrier and the brain concentration reached a maximum, which was about 10 times higher in knockout mice than in normal mice, 1 hr after administration. The brain concentration in normal mice thereafter decreased slowly, whereas in knockout mice, an extremely high concentration was maintained for 24 hr. CONCLUSIONS: The pharmacokinetic behavior of FK506 in the tissue distribution is related with the function of P-glycoprotein encoded by the mdrla gene. The brain distribution of FK506 is dominated by the P-glycoprotein-mediated drug efflux and presumably also by the binding to FK-binding proteins (immunophilins) in the brain.

[Adenocarcinoma of the bladder producing carcinoembryonic antigen and carbohydrate antigen 19-9: report of a case].

We report a case of primary adenocarcinoma of the bladder producing carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9). An 85-year old woman presented with gross hematuria. Computerized tomography and ultrasonography demonstrated a mass on the urinary bladder. After clinical examination, she was diagnosed with locally invasive bladder cancer. Serum CEA and CA19-9 levels were increased to 8.9 ng/ml (normal < 2.5) and 870 U/ml (normal < 37), respectively. Simple cystectomy and bilateral cutaneous ureterostomy were performed. The histopathological diagnosis was well-differentiated adenocarcinoma of the bladder with invasion through the entire muscle layer. Postoperatively, the serum CEA and CA19-9 levels decreased to 5.4 ng/ml and 53 U/ml, respectively, and both CEA and CA19-9 were detected by immunohistochemical examination of the tumor.

Inhibition by protein kinase C inhibitor of expression of leukocyte function-associated antigen-1 molecules in rat hepatoma AH66F cells.

To examine the mechanism of inhibition by protein kinase C (PKC) inhibitors of the adhesion of highly malignant hepatoma AH66F cells to the mesentery-derived mesothelial cell (M-cell) layer through leukocyte function-associated antigen-1 (LFA-1)/intercellular adhesion molecule-1, the effects of a PKC inhibitor, NA-382, on the expression of LFA-1 molecules in AH66F cells were examined and compared with those in thymocytes from normal rats. NA-382 inhibited the adhesion of AH66F cells to the M-cell layer and the expression of LFA-1 on the membrane of the hepatoma cells after treatment for more than 24 h. It was confirmed that AH66F cells express similar mRNAs for LFA-1 subunits to those of thymocytes, and their levels were also decreased after treatment with NA-382. On the other hand, the LFA-1 mediated adhesion and the expression of both protein and mRNA for LFA-1 subunits in thymocytes were not changed by the PKC inhibitor. These results suggest that the expression of LFA-1 molecules in AH66F cells may be regulated by PKC via quite different mechanisms from those in normal lymphocytes.

Dual regulation of alkaline phosphatase activity by calcitonin in porcine kidney cells.

Intracellular signal transduction for regulation of alkaline phosphatase (ALP) activity in renal epithelial cells treated with calcitonin is not yet completely understood, although it is known that calcitonin receptors couple to cyclic AMP-dependent protein kinase (PKA) and calcium/phospholipid-dependent protein kinase (PKC). Salmon calcitonin increased the cyclic AMP content in LLC-PK1 porcine kidney cells in a concentration-dependent manner. When the confluent cells were incubated for 47 h after a 1 h-pulse exposure or continuously exposed to calcitonin and forskolin for 48 h, ALP activity in the cells was increased by calcitonin about 2-fold compared with the basal activity at the maximum level but was not dependent on the exposure time; it was markedly increased by forskolin in parallel with the exposure time. The increase in activity produced by calcitonin was abolished by a PKA inhibitor H-89, and, in contrast, potentiated by a PKC inhibitor, NA-382 to near the forskolin-induced level. These results indicate that calcitonin exerts a dual-regulation of ALP activity in LLC-PK1 cells, positively through the PKA pathway and negatively through PKC.

Cytotoxic components of bardanae fructus (goboshi).

Bardanae Furctus (Goboshi) extract showed potent in vitro cytotoxicity and in vivo antitumor activity against human hepatoma HepG2 cells and mouse sarcoma 180 cells, respectively. In this study, the cytotoxicities of four fractions and three major components (arctiin, arctigenin, and chlorogenic acid) isolated from Goboshi extract were examined. Arctiin and arctigenin, which were contained in the ethylacetate fraction and n-butanol fraction, respectively, showed strong cytotoxicity against HepG2 cells, but little toxicity against Chang liver cells. Chlorogenic acid isolated from the water fraction did not affect the viability of these cells. The cytotoxicity of arctigenin against Chang liver cells was markedly potentiated by treatment with glutathione (GSH) synthesis inhibitor, L-buthionine-(S,R)-sulfoximine (BSO). On the other hand, in HepG2 cells, the cytotoxicity of arctigenin was hardly changed by BSO. The cytotoxicity of arctigenin against HepG2 cells increased in an exposure-time dependent manner. These characteristics of arctigenin were similar to those of Goboshi extract, as previously observed. We therefore conclude that the principal cytotoxic components of Goboshi extract are arctiin and its aglycone arctigenin.

In vivo cisplatin resistance of rat ascites hepatoma AH66.

The rat ascites hepatoma AH66 cell line had higher resistance to cisplatin than a variant AH66F cell line in in vivo experiments, though in vitro sensitivities to cisplatin of both cell lines were similar in a medium containing 5% fetal calf serum (FCS). When AH cells were cultured in the medium containing 5% ascites fluid (ASF), the sensitivity of AH66 cells to cisplatin was significantly lower than that in the medium containing 5% FCS, but the sensitivity of AH66F cells was almost same in the case of 5% FCS. The metallothionein (MT) contents in AH66 cells, but not in AH66F cells, increased with the incubation time in the medium containing ASF. MT in AH66 cells was also induced by treatment with zinc ion, but the induction in AH66F cells was very low. These results indicate that AH66 cells have a high ability to induce MT and thereby may acquire in vivo resistance to cisplatin.

Phenotypic characteristics of Epstein-Barr-virus-associated gastric carcinomas.

We identified Epstein-Barr-virus(EBV)-associated gastric carcinomas by in situ hybridization that targets EBV-encoded small RNA, and investigated their phenotypic characteristics by mucin histochemistry. Out of 132 gastric carcinomas, 15 were EBV-positive, and they were exclusively located at the proximal part of the stomach; 8 were early and 7 were advanced carninomas. Out of the 15 EBV-positive carcinomas, 10 were so-called gastric carcinoma with lymphoid stroma (GCLS); 4 of the other 5 tumours were moderately differentiated adenocarcinoma (tub 2) showing a "lace pattern". This pattern was also seen in the intramucosal parts of 6 (3 early and 3 advanced) GCLS. As for the mucin phenotype of the cancer cells, the gastric type was predominant in the EBV-positive tumours, while the EBV-negative tumours, even with the lace pattern or GCLS, showed phenotypes other than the gastric type (intestinal or mixture of intestinal and gastric). It was inferred that EBV-positive gastric carcinomas show tub 2 with the lace pattern and gastric phenotype in earlier stages, and become GCLS during the tumour progression, but the gastric phenotype tends to remain throughout the history.

Absorptive-mediated endocytosis of an adrenocorticotropic hormone (ACTH) analogue, ebiratide, into the blood-brain barrier: studies with monolayers of primary cultured bovine brain capillary endothelial cells.

The internalization of a neuromodulatory adrenocorticotropic hormone (ACTH) analogue, [125I]ebiratide (H-Met(O2)-Glu[125I]His-Phe-D-Lys-Phe-NH(CH2)2NH2), was examined in cultured monolayers of bovine brain capillary endothelial cells (BCEC). HPLC analysis of the incubation solution showed that [125I]ebiratide was not metabolized during the incubation with BCEC. The acid-resistant binding of [125I]ebiratide to BCEC increased with time for 120 min and showed a significant dependence on temperature and medium osmolarity. Pretreatment of BCEC with dansylcadaverine or phenylarsine oxide, endocytosis inhibitors, and 2,4-dinitrophenol, a metabolic inhibitor, decreased significantly the acid-resistant binding of [125I]ebiratide. The acid-resistant binding of [125I]ebiratide was saturable in the presence of unlabeled ebiratide (100 nM-1 mM). The maximal internalization capacity (Bmax) at 30 min was 7.96 +/- 3.27 pmol/mg of protein with a half-saturation constant (Kd) of 15.9 +/- 6.4 microM. The acid-resistant binding was inhibited by basic peptides such as poly-L-lysine, protamine, histone, and ACTH but was not inhibited by poly-L-glutamic acid, insulin, or transferrin. These results confirmed that ebiratide is transported through the blood-brain barrier via an absorptive-mediated endocytosis.

P-glycoprotein as the drug efflux pump in primary cultured bovine brain capillary endothelial cells.

The expression of a functional P-glycoprotein (P-gp) which pumps drugs out of brain capillary endothelial cells (BCEC) into blood was studied by evaluating the steady-state uptake and efflux of vincristine (VCR) by primary cultured bovine BCEC. The steady-state uptake of VCR was increased in the presence of metabolic inhibitors, and an anti-P-gp monoclonal antibody, MRK16, as well as verapamil and steroid hormones which are known to reverse multidrug resistance in tumor cells. Furthermore, efflux of VCR from BCEC was inhibited by verapamil. By immunohistochemistry, P-gp was localized at the luminal side of the capillary endothelial cells in both gray matter of bovine brain and primary cultured BCEC. These data suggest that P-gp functions as a drug efflux pump at the luminal side of BCEC and regulates the transfer of certain lipophilic drugs from the blood into the brain.

Transport of monocarboxylic acids at the blood-brain barrier: studies with monolayers of primary cultured bovine brain capillary endothelial cells.

The kinetics and mechanism of the transport of monocarboxylic acids (MCAs) were studied by using primary cultured bovine brain capillary endothelial cells. Concentration-dependent uptake of acetic acid was observed, and the kinetic parameters were estimated as follows: the Michaelis constant, Kt, was 3.41 +/- 1.87 mM, the maximum uptake rate, Jmax, was 144.7 +/- 55.7 nmol/mg of protein/min and the nonsaturable first-order rate constant, Kd, was 6.66 +/- 1.98 microliters/mg of protein/min. At medium pH below 7.0, the uptake rate of [3H]acetic acid increased markedly with decreasing medium pH, whereas pH-independent uptake was observed in the presence of 10 mM acetic acid. An energy requirement for [3H]acetic acid uptake was also demonstrated, because metabolic inhibitors (2,4-dinitrophenol and rotenone) reduced significantly the uptake rate (P less than .05). Carbonylcyanide-p-trifluoro-methoxyphenylhydrazone, a protonophore, inhibited significantly the uptake of [3H]acetic acid at medium pH of 5.0 and 6.0, whereas 4,4'-diisothiocyanostilben-2,2'-disulfonic acid did not. Several MCAs inhibited significantly the uptake rate of [3H]acetic acid, whereas di- and tricarboxylic acids did not. The uptake of [3H]acetic acid was competitively inhibited by salicylic acid, with an inhibition constant, Ki, of 3.60 mM, suggesting a common transport system between acetic acid and salicylic acid. Moreover, at the medium pH of 7.4, salicylic acid and valproic acid inhibited significantly the uptake of [3H]acetic acid, demonstrating that the transport of MCA drugs could also be ascribed to the MCA transport system at the physiologic pH.(ABSTRACT TRUNCATED AT 250 WORDS)