Hiroshi Mori

Renal cell carcinoma in dialysis patients: a single center experience.

Aim: Renal cell carcinoma (RCC) is a life‐threatening complication of end‐stage renal disease with an unclear pathogenesis. We evaluated RCC developing in patients undergoing dialysis.

An immunohistochemical study of hepatic atypical adenomatous hyperplasia, hepatocellular carcinoma, and cholangiocarcinoma with α‐fetoprotein, carcinoembryonic antigen, CA19‐9, epithelial membrane antigen, and cytokeratins 18 and 19

Eight hepatic atypical adenomatous hyperplasias (AH), 30 hepatocellular carcinomas (HCC) consisting of 11 well‐, 13 moderately and six poorly differentiated HCC, and 10 intrahepatic cholangiocarcinomas (CC) were investigated immunohistochemically with anti‐α‐fetoprotein (AFP), carcinoembryonic antigen (CEA), CA19‐9, epithelial membrane antigen (EMA), and cytokeratins (CK) 18 and 19 antibodies. Immunostaining was regarded as positive when more than 5% of cells were stained. α‐Fetoprotein was positive, although focally, in five (17%) of 30 HCC but negative in all AH and CC. Carcinoembryonic antigen (polyclonal antibody) did not stain the cytoplasm of all AH and HCC, but stained two (25%) of eight AH and 10 (33%) of 30 HCC in a bile canalicular staining manner. Carcinoembryonic antigen showed intracytoplasmic or luminal border staining in six (60%) of 10 CC. CA19‐9 was negative in all AH and HCC, while six (60%) of 10 CC were positive for CA19‐9. Epithelial membrane antigen was positive in one (13%) of eight AH, seven (23%) of 30 HCC and in all 10 cases of CC. Cytokeratin 18 was positive in all AH, HCC and CC. Cytokeratin 19 was negative in both AH and HCC, whereas it stained the cytoplasm of tumor cells in all CC diffusely and intensely. These results suggest that immunostaining of AFP, CEA, CA19‐9, EMA, CK18 and CK19 are not useful in the differential diagnosis between AH and well‐differentiated HCC, and that CK19 is the most suitable reagent for the differential diagnosis between HCC and CC.

ATP regulation of ciliary beat frequency in rat tracheal and distal airway epithelium

Ciliary beat frequency (CBF) was measured by video‐optical microscopy in rat tracheal and distal airway ciliary cells using a slice preparation. In tracheal ciliary cells (tracheal slice), ATP or 2‐methylthio ATP (MeSATP) increased CBF, which was inhibited by suramin (100 μm, an inhibitor of purinergic receptor). Ionomycin (5 μm) or thapsigargin (2 μm) increased CBF similarly. Ca2+‐free solution or addition of Ni2+ (1 mm) decreased CBF gradually by approximately 25% and subsequent stimulation with ATP (10 μm) increased CBF transiently. The purinergic agonist experiments demonstrated that ATP increases CBF in tracheal ciliary cells via both P2X and P2Y receptors. ATP increased the intracellular calcium concentration ([Ca2+]i) in tracheal ciliary cells. However, in distal airway ciliary cells (lung slice), ATP did not increase CBF and [Ca2+]i, although a Ca2+‐free solution decreased CBF, and ionomycin (5 μm) or thapsigargin (2 μm) increased it. Moreover, acetylcholine (100 μm) did not increase CBF in distal airway ciliary cells, although it increased CBF in tracheal ciliary cells. Terbutaline (10 μm), a selective β2‐adrenergic agonist, increased CBF in both tracheal and distal airway ciliary cells. These observations suggest that the Ca2+‐mobilization mechanisms via purinergic or muscarinic receptors of the distal airway ciliary cell may be different from those of the tracheal ciliary cell. In conclusion, the CBF increase is differently regulated in the tracheal and distal airway epithelia of the rat.

ULTRASTRUCTURAL STUDY ON CYTOTOXIC EFFECTS OF CYCLOSPORINE A IN SPERMIOGENESIS IN RATS

Cyclosporine A (CsA) is known to have testicular toxicity, leading to male infertility. The occurrence of numerous anomalous spermatids and residual bodies in the epididymal ducts of rats treated with CsA was observed in our previous studies. To examine the fine structural changes of impaired spermiogenesis induced by CsA, rats were treated s.c. with 40u2009mg/kg/day CsA for 2 weeks. Desquamation of round spermatids still surrounded by a slender Sertoli cell cytoplasm was occasionally observed. A prominent change in the seminiferous tubules was an occurrence of numerous residual bodies containing one or more flagella. They were not phagocytosed by the Sertoli cell, and accumulated in the epididymal ducts. Cellular components in the epididymal lumen included degenerating round spermatids and abnormal spermatozoa and residual bodies. Although separation of the head from the tail of the spermatozoa was rarely seen, various kinds of abnormalities of flagella were frequently encountered; compact aggregation of numerous flagella in a single spermatozoon or residual body, disarrangement of mitochondrial or fibrous sheath with outer dense fibers, and fusion of flagella with a single intervening mitochondrial layer. These findings indicate that CsA gives rise to toxic effects on the spermiogenesis by impairing directly the spermiogenic cell development and by impeding Sertoli cell function including a reduction of its phagocytic activity.

A case of malignant lymphoma of the ovary manifesting like an advanced ovarian cancer

BACKGROUNDnIn recent years, true primary ovarian lymphoma has been considered to carry a favorable prognosis, although most studies of supposedly primary ovarian lymphoma have reported a poor outcome.nnnCASEnA 47-year-old woman presented with signs and symptoms suggestive of an advanced ovarian cancer. Ultrasonography and magnetic resonance imaging revealed bilateral abdominal tumors, each measuring 10 cm in diameter, thickened omentum, and a large amount of ascitic fluid, but no enlarged lymph nodes. The diagnosis of malignant lymphoma was established from the biopsy specimen after exploratory laparotomy. Six years following chemotherapy, the patient is alive and disease free without additional surgery.nnnCONCLUSIONnThe prognosis of ovarian lymphoma was evaluated according to clinical stage, modality of onset, histologic type, and phenotype. It remains controversial whether this case can be considered truly primary ovarian lymphoma and not merely a localized initial manifestation of a generalized disease. But if this case of advanced ovarian lymphoma were not primary, it could still be managed successfully with chemotherapy appropriate for the specific histology.

Expression of matrix metalloproteinases, tissue inhibitors of metalloproteinase, collagens, and Ki67 antigen in pleural malignant mesothelioma : an immunohistochemical and electron microscopic study

Because matrix metalloproteinases (MMPs) degrade extracellular matrix, including basement membrane, and because tissue inhibitors of MMP (TIMPs) suppress MMP activities, MMPs and TIMPs are considered to play important roles in invasion and metastasis in many malignancies. We examined immunohistochemically the expression of MMPs (MMP-1, -2, -3, -7, and -9), TIMPs (TIMP-1 and -2), and collagens (types I, III, and IV) in 16 patients with pleural malignant mesothelioma (PMM; 8 with the epithelial, 4 with the sarcomatous, and 4 with the biphasic type). Electron microscopy revealed that the tumor cells in all types possessed the characteristics of malignant mesotheliomas, including numerous microvilli and moderate amounts of intermediate filaments. Basement lamina was present only focally. The proliferative Ki67 index was at a high level, compared with values reported in various other malignancies. Positive staining for MMP-1 was observed in most tumor cells in all 16 patients (100%). MMP-2 was expressed in most tumor cells in 2 patients (13%). In contrast, MMP-3, -7, and -9 were not detected in any PMM. TIMP-1 and TIMP-2 were expressed in 3 patients (19%) and 2 patients (13%), respectively. The stromal cells were simultaneously positive for MMPs or TIMPs in the patients whose tumor parenchymal cells were positive for each enzyme. These results indicate that the expression of MMP-1 and MMP-2 may be related to PMM invasion and spread. In particular, as MMP-1 was overexpressed in contrast to the lower expression of TIMP-1, MMP-1 is strongly suggested to play an important role in PMM invasion by degrading the tumor stroma. In spite of general agreement that epithelial-type PMM has a better prognosis than other types, there was no significant difference in the Ki67 index among the histological types of PMM.

Endoscopic stent placement for treatment of secondary bilateral occlusion of the Monro foramina following endoscopic third ventriculostomy in a patient with aqueductal stenosis

Nontumoral bilateral occlusion of the Monro foramina is a rare clinical condition. Treatment includes shunt placement, endoscopic procedures, or both. The authors describe the case of a 22-year-old woman who had previously undergone placement of a ventriculoperitoneal shunt via a right frontal approach for management of triventricular dilation due to aqueductal stenosis. Six years postoperatively she presented with right-sided slit-ventricle syndrome and stenosis of the right Monro foramen, which was treated with an endoscopic third ventriculostomy and fenestration of the septum pellucidum. Two years later she presented with bilateral lateral ventricular dilation. Inspection of the right lateral ventricle with a fiberscope revealed occlusion of the septum pellucidum fenestration; on observation, the right Monro foramen was covered by thick, tough granulation tissue and the left was occluded by thin membranous tissue. Repeated fenestration of the septum pellucidum and left Monro foraminoplasty were therefore performed by perforating this thin tissue. A stent was then introduced into the third ventricle via the right lateral ventricle, the fenestration in the septum pellucidum, and the left Monro foramen. The authors note that fiberscopes are in general more maneuverable than rigid endoscopes and conclude that they are particularly useful for the treatment of this type of hydrocephalus.

β2‐adrenergic regulation of ciliary beat frequency in rat bronchiolar epithelium: potentiation by isosmotic cell shrinkage

Single bronchiolar ciliary cells were isolated from rat lungs. The β2‐adrenergic regulation of ciliary beat frequency (CBF) was studied using video‐optical microscopy. Terbutaline (a β2‐adrenergic agonist) increased CBF in a dose‐dependent manner, and it also decreased the volume of the ciliary cells. These terbutaline actions were inhibited by a PKA inhibitor (H‐89) and mimicked by forskolin, IBMX and DBcAMP. Ion transport inhibitors were used to isosmotically manipulate the volume of the terbutaline‐stimulated bronchiolar ciliary cells. Amiloride (1 μm) and bumetanide (20 μm) potentiated cell shrinkage and the CBF increase, and they shifted the terbutaline dose–response curve to the lower‐concentration side. Quinidine (500 μm), in contrast, increased cell volume and suppressed the CBF increase. Moreover, a KCl solution containing amiloride (1 μm) and strophanthidin (100 μm) increased cell volume and suppressed the CBF increase, and then the subsequent removal of either amiloride or strophanthidin decreased cell volume and further increased CBF. NPPB (10 μm) or glybenclamide (200 μm) had no effect on the action of terbutaline. Thus, in terbutaline‐stimulated ciliary cells, cell shrinkage enhances the CBF increase; in contrast, cell swelling suppresses it. However, the results of direct manupulation of cell volume by applying osmotic stresses (hyperosmotic shrinkage or hyposmotic swelling) were the opposite of the findings of the isosmotic experiments: hyposmotic cell swelling enhanced the CBF increase, while isosmotic swelling suppressed it. These results suggest that isosmotic and non‐isosmotic volume changes in terbutaline‐stimulated bronchiolar ciliary cells may trigger different signalling pathways. In conclusion, terbutaline increases CBF and decreases the volume of rat bronchiolar ciliary cells via cAMP accumulation under isosmotic conditions, and the isosmotic cell shrinkage enhances the CBF increase by increasing cAMP sensitivity.

Quantification of immunohistochemistry using an image analyser: correlation with hormone concentrations in pituitary adenomas

SummaryThe usefulness of immunohistochemistry is usually confined to qualitative analysis. Quantitative evaluation is not performed. At best, the number of immunopositive cells and the immunointensities are recorded as several grades, to which a strict categorization may be applied by individual examiners, but these categorizations are not standardized. We have attempted to quantify immunohistochemical observations using an image analyser. Sections from rat pituitary adenomas secreting prolactin and growth hormone were immunostained for these hormones with either immunogold silver or avidin-biotinylated peroxidase complex (ABC) methods. The number of immunopositive cells were counted by eye on specimens stained with the ABC method. In sections stained by an immunogold-silver technique, an immunopositive area was measured at several immunointensity ranges, to which certain points were allotted. Immunohistochemical values obtained by summing the products of the immunopositive area and intensity points at each range were correlated with concentrations of hormones in adenoma tissues measured by radioimmunoassay. A high correlation between the immunohistochemical values and hormone concentrations were shown for both prolactin and growth hormone, in contrast to a low correlation between the number of immunopositive cells counted by eye and the hormone concentrations. These findings indicate that the immunohistochemical observations can be quantified using the image analyser to the extent that they can be substituted, albeit roughly, for the hormone concentrations measured biochemically.

Ciliary beat frequency controlled by oestradiol and progesterone during ovarian cycle in guinea‐pig Fallopian tube

The ciliary beat frequency (CBF) of guinea‐pig fimbria during the ovarian cycle was measured by video microscopy using a high‐speed camera (500 Hz). In the follicular phase, with increasing concentrations of β‐oestradiol ([βE2]) and a low concentration of progesterone ([PRG]), CBF increased from 13.5 to 16 Hz. In the ovulatory phase, with further increase of [βE2], CBF decreased gradually from 16 to 13.5 Hz. In the early luteal phase, with low [PRG] and [βE2], CBF increased to ∼17 Hz; however, in the middle luteal phase, with increasing [PRG], CBF decreased (∼12 Hz), and in the late luteal phase, with decreasing [PRG], CBF increased to ∼15 Hz. Then, in the resting phase, with low [βE2] and [PRG], CBF decreased immediately to ∼14 Hz. The CBF of the fimbria was measured in guinea‐pigs treated with β‐oestradiol benzoate (βE2B) or medroxyprogesterone (mPRG). A low dose of βE2B increased CBF to ∼14.5 Hz, whereas a high dose decreased it to ∼11 Hz. A βE2 receptor blocker, ICI‐182,780, abolished the βE2B‐induced CBF changes and maintained CBF at ∼12.0 Hz. Medroxyprogesterone decreased CBF to ∼12.5 Hz, and mifepristone (a PRG receptor blocker) abolished the mPRG‐induced CBF decrease and maintained CBF at ∼15 Hz. The addition of both blockers increased CBF to ∼18 Hz, suggesting that activation of βE2 or PRG receptors decreases the CBF of the fimbria. In conclusion, a moderate [βE2] increase maintains a high CBF (15.5 Hz) in the follicular phase, and then further [βE2] increase decreases CBF to 13.5 Hz in the ovulatory phase. In the early and late luteal phase, low [βE2] and [PRG] increase CBF to 17 and 15 Hz, respectively, and in the middle luteal phase a high [PRG] decreases CBF (to ∼12 Hz). Thus, the CBF of the fimbria was controlled by signals via βE2 and PRG receptors in guinea‐pigs.