A case of bilateral middle-ear squamous cell carcinoma.

Only eight cases of bilateral middle-ear squamous cell carcinoma (SCC) have been reported to date. We present the case of a 75-year-old male with bilateral middle-ear SCC and review the previously reported cases. The patient was diagnosed as having moderately-differentiated SCC in the left middle ear in February 1995 and well-differentiated SCC in the right middle ear in September 1997. He initially received radiation therapy with (60)Co pendulum (64 Gy) in the left ear and was subsequently treated by Liniac irradiation (50 Gy) in the right ear. He has now been followed up at our ENT clinic for 29 months without vertigo or facial nerve palsy since the second radiation therapy. Although he has a residual tumour in the right middle ear invading the middle cranial fossa dura, no sign of recurrence has been detected in the left ear.

Primary low-grade MALT lymphoma of the gallbladder.

We report a case of primary low-grade B-cell lymphoma of the mucosa-associated lymphoid tissue (low-grade MALT lymphoma) in the gallbladder. A 58-year-old woman suspected of gallbladder carcinoma underwent laparoscopic cholecystectomy. Microscopic examination of the gallbladder demonstrated lymphoid cell infiltration forming lymphoid follicles with hyperplastic secondary follicles. The surrounding monocytoid B cells and centrocyte-like cells selectively infiltrated the crypt epithelium forming lympho-epithelial lesions. Plasma cells were also noted beneath the mucosal epithelium. Bile culture revealed the Gram-negative bacilli Enterococcus faecalis and Morganella morganii. Immunoglobulin heavy chain gene rearrangement was confirmed using polymerase chain reaction (PCR) and oligoclonal lymphoid proliferations was detected. Because autoimmune diseases, or chronic inflammatory disorders, seem to correlate with the occurrence of MALT lymphoma, Gram-negative bacterial infection could also be considered as a prodrome of MALT lymphoma of the gallbladder.

Intravenous injection of cycloheximide induces apoptosis and up-regulates p53 and Fas receptor expression in the rat liver in vivo.

A single administration of protein synthesis inhibitor, cycloheximide (CHX) induces apoptosis of hepatocytes in vivo. We investigated the underlying mechanisms of this phenomenon and the role of p53 and Fas receptor using terminal dUTP nick end labeling (TUNEL), quantitative reverse transcription polymerase chain reaction and immunohistochemistry. Rat liver tissue specimens were obtained at different time intervals after injection of CHX. The proportion of TUNEL-positive apoptotic hepatocytes increased with time and reached a plateau at 2.5h after the injection. The p53 and Fas receptor mRNAs and the proportion of immunoreactive p53-positive and Fas receptor-positive hepatocytes increased markedly with time from 1h after the administration. Since the time course of increased proportion of apoptotic hepatocytes does not parallel that of p53- or Fas receptor-positive hepatocytes and apoptotic hepatocytes appeared prior to up-regulation of p53 and Fas receptor expression, it is likely that the enhanced expression of p53 and Fas receptor is not involved directly in CHX-induced apoptosis of hepatocytes in vivo. Rats injected with a single intravenous dose of CHX, however, provide a simple and useful model for investigating the apoptotic machinery and the molecular mechanism of transcriptional up-regulation of p53 and Fas receptor in hepatocytes.

Apoptosis in the aging process.

Although many hypotheses have been proposed to explain the aging process, the exact mechanisms are not well defined. Recent accumulating evidence indicates that dysregulation of the apoptotic process may be involved in some aging processes; however, it is still debatable how exactly apoptosis is expressed during aging in vivo. In this review, we discuss recent findings related to apoptosis of individual organs during aging and their significance. We demonstrate that aging enhances apoptosis and susceptibility to apoptosis in several types of intact cells. In contrast, in certain genetically damaged, initiated, and preneoplastic cells, aging suppresses these age-associated apoptotic changes. In various cells, apoptosis enhances the elimination of damaged and dysfunctional cells presumably caused by oxidative stress, glycation, and DNA damage. In these cases, the incidence of apoptosis correlates with the level of accumulated injury. It is concluded that apoptosis plays an important role in the aging process and tumorigenesis in vivo probably as an inherent protective mechanism against age-associated tumorigenesis.

Dietary restriction reduces hepatocyte proliferation and enhances p53 expression but does not increase apoptosis in normal rats during development.

Dietary restriction (DR) retards physical growth, resulting in small body size, reduced liver weight and reduced number of hepatocytes in rats. We examined the effects of DR on proliferation and apoptosis of hepatocytes during development and explained these changes subcellularly using immunohistochemistry for bromodeoxyuridine (BrdU) incorporation, proliferating cell nuclear antigen (PCNA) and p53, terminal dUTP nick end labeling (TUNEL) and quantitative reverse transcription-polymerase chain reaction (RT-PCR) for TGFalpha, TGFbeta1, p53, Fas and TNF receptor (TNFr). Tissue samples included the livers of 3-month-old male Fischer 344 rats fed ad libitum (AL) or on 70% DR. DR significantly reduced the proportions of BrdU-positive and PCNA-strongly-positive hepatocytes compared with AL rats but not the proportions of PCNA-positive hepatocytes and TUNEL-positive hepatocytes. On the other hand, DR enhanced the expression of p53 and Fas mRNAs but failed to influence the expression of TGFalpha, TGFbeta1 and TNFr mRNAs. Moreover, DR significantly increased the proportion of p53-positive hepatocytes. Our findings suggest that DR suppresses the proliferation of hepatocytes, resulting in a reduced number of hepatocytes during development. This process may be mediated by overexpression of p53 suppressor gene. While DR accelerates the expression of Fas antigen, this result does not influence the rate of apoptosis of hepatocytes under physiological conditions.

Granulomatous lobular mastitis: difficulty of diagnosis.

We report a case of a rare inflammatory disease, granulomatous lobular mastitis. Two weeks prior to admission the patient, a 43 year-old woman, (gravida 1, para 1) had noticed a left breast mass associated with tenderness. Palpation, gross inspection, and clinical examination, as well as the rapid growth of the mass lesion led us to believe that it was highly suspicious of malignant neoplasm. Mammography, ultrasonography, and computed tomography did not differentiate it from a malignant neoplasm. Aspiration cytology revealed an inflammatory lesion with a few clusters of epithelial cells it was diagnosed as borderline malignancy(class III) by a prudent pathologist, and thus mastectomy was performed. However, the final histologi-cal diagnosis was granulomatous lobular mastitis with no evidence of malignancy. As the clinical manifestations of granulomatous mastitis are similar to those of mammary carcinoma and, as it is an inflammatory lesion of uncertain etiology and pathogenesis, it has often been mistaken clinically for carcinoma and treated as such. Our review of the literature indicated that granulomatous mastitis most often occurs in young patients with a history of childbirth or oral contraceptive usage. Recurrence was documented in 38% of patients, and, accordingly long-term follow-up by aspiration cytology, complete resection, and adequate drug treatment with corticosteroids are recommended.

Malignant mesothelioma of the tunica vaginalis testis: report of a case.

We describe herein the case of a 68-year-old man with malignant mesothelioma of the tunica vaginalis testis. The pathological diagnosis was based upon the clinical findings, gross and microscopic morphology, and special stains. Malignant mesothelioma is a rare tumor associated with asbestos exposure that can be effectively treated with orchidectomy via an inguinal approach.

Clinical experiences of microsurgical side-to-end epididymovasostomy for epididymal obstruction.

BACKGROUND: Some surgical treatments are performed for obstructive azoospermia in urology and good results have been reported. Of 61 azoospermic patients who visited our department of urology, nine were diagnosed as having epididymal obstruction of unknown etiology. METHODS: We describe nine consecutive side-to-end epididymovasostomy procedures performed on these patients. These procedures are microsurgical two-layer anastomosis. RESULTS: Of the nine men, five (55.6%) had sperm in the ejaculate postoperatively and, up until publication, the pregnancy rate was 33.3% (three of nine). CONCLUSIONS: These results suggest that reconstruction of the seminal tract should be considered first for obstructive azoospermia.

Life-long caloric restriction suppresses age-associated Fas expression in the Fischer 344 rat kidney.

It has been shown that the expression of Fas is substantially increased in the aging process in various organs, but its role in the aging kidney is not yet clear. In this study, the expression of Fas in the kidneys of 6- and 24-month-old male Fischer 344 rats fed ad libitum was studied by using quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. In addition, possible effects of life-long caloric restriction (30% as those of ad libitum fed group) in the expression of Fas were also studied in 6- and 24-month-old rat kidneys. Kidneys obtained from 24-month-old ad libitum fed rats showed glomerulosclerosis with marked tubulointerstitial damage including interstitial fibrosis, while in the kidneys of 24-month-old calorie-restricted rats, renal damage was remarkedly less than that noted in 24-month-old ad libitum fed rats kidneys. RT-PCR and immunohistochemical analysis showed an increased expression of Fas in both mRNA and protein level in 24-month-old rat kidneys; life-long caloric restriction significantly reduces renal expression of Fas. Our results suggest that increased expression of Fas is associated with age-related renal damage and that life-long diet-restricted alteration of its expression is associated with the modulation of age-associated renal structural damage.

Malignant peripheral nerve sheath tumors developing multifocally in the central nervous system in a patient with neurofibromatosis type 2.

We describe autopsy findings of multifocal malignant peripheral nerve sheath tumors (MPNSTs) appearing in the central nervous system in a 45-year-old Japanese female with neurofibromatosis type 2. Multiple MPNSTs were detected in both III and VIII, left IV, and V cranial nerves, and a number of nerve roots of the spinal cord. Neurofibromata were on the other hand evident on some nerve roots of the spinal cord and femoral and sciatic nerves. Our results suggest that a mutation of p53 gene may have played a role in the malignant transformation of nerve tumors in this patient since p53 protein was immunohistochemically detected in MPNST cells but not in tumor cells of the neurofibromata.

Advanced glycosylation end products in adrenal lipofuscin.

The present study examined the presence of advanced glycosylation end products (AGEs) in lipofuscin present in the brain and adrenal gland of aging rats by immunohistochemistry using antibodies raised against AGEs. Lipofuscin identified as yellow to brown granules emitting bright yellow to orange autofluorescence with ultraviolet light were detected in cortical neurons, cerebellar Purkinje cells, and adrenal cells in the inner part of the zona reticularis. However, none of the antibodies visualized lipofuscin in these areas. The outer part of the zona reticularis contained yellow granules emitting a faint orange autofluorescence. These granules were immunostained by an antibody that reacted with AGEs structures unrelated to the carboxymethyllysine moiety. Newly formed adrenal cortical cells are thought to migrate from the outer layer to the inner layer of the zona reticularis. Therefore, our results suggest that glycosylation-related processes are involved in lipofuscinogenesis, at least in its early stage, in the adrenal zona reticularis.

Allergic granulomatous angitis (Churg-Strauss syndrome) with multiple intestinal fistulas.

A 40-yr-old man who had a known diagnosis of allergic granulomatous angitis (Churg-Strauss syndrome) and had been on steroids was found to have a stone in the common bile duct. At surgery, multiple internal fistulas were found in the small bowel. Cholecystectomy, removal of the stone in the common bile duct, and resection of the small bowel because of fistulas were performed. To our knowledge, the formation of an intestinal fistula has not been reported as a clinical manifestation of allergic granulomatous angitis. This rare condition occurs in the terminal stage of this disease.

Aging accelerates but life-long dietary restriction suppresses apoptosis-related Fas expression on hepatocytes.

Aging enhances apoptosis of hepatocytes under normal physiological conditions and increases the susceptibility of hepatocytes to apoptosis whereas life-long dietary restriction suppresses the age-enhanced susceptibility to apoptosis. We examined the subcellular mechanisms of the age-associated changes and effect of dietary restriction using quantitative reverse transcription polymerase chain reaction and immunohistochemistry for Fas in the livers of 6- and 24-month-old male Fischer 344 rats fed ad libitum or 70% diet restricted. We also analyzed the level of ordinary and variant forms of Fas mRNA. The ordinary form of Fas mRNA, but not the variant form of Fas mRNA, significantly increased with age. Dietary restriction significantly suppressed the ordinary form of Fas mRNA in advanced age. Aging enhanced Fas immunoreactivity in the hyperplastic bile epithelium and hepatocytes whereas dietary restriction suppressed it. Our findings indicate that Fas protein, particularly the ordinary form of Fas, is involved in age-associated apoptosis of hepatocytes. Fas overexpression in advanced age may explain the age-enhanced susceptibility to apoptosis. Our results also suggest that dietary restriction suppresses Fas overexpression, resulting in a reduction of the age-enhanced susceptibility to apoptosis.

In vivo retrovirus-mediated herpes simplex virus thymidine kinase gene therapy approach for adult T cell leukemia in a rat model.

We have previously demonstrated that human T-lymphotropic virus type I (HTLV-I) tax-expressing human T cell lines are selectively eliminated in the presence of aciclovir, using a retroviral vector carrying the herpes simplex virus thymidine kinase (HSV TK) gene under the control of the long terminal repeat (LTR) of HTLV-I. Based on these findings in vitro, we investigated whether this system could also be effective in vivo, using a rat model. Following infection of the HTLV-I-transformed and tax-expressing rat T cell line TARS-1 with this retrovirus (LNLTK virus), high levels of HSV TK expression were observed and resulted in increased susceptibility to ganciclovir (GCV). Tumors were generated by subcutaneous injection of TARS-1 in newborn syngeneic WKA/H rats. While the tumors derived from infected TARS-1 cells with control virus, as well as uninfected cells, continued to grow in all the rats with or without administration of GCV, those derived from LNLTK-infected cells exhibited dramatic regression upon GCV treatment. These results indicate that the HTLV-I LTR-HSV TK system also causes selective elimination of HTLV-I-transformed, tax-expressing T cells in vivo. Therefore, our present study may provide a rationale for clinical gene therapy against adult T cell leukemia.

Physical activity as a factor in the action of dietary restriction on aging: effects in Fischer 344 rats.

Dietary restriction (DR) slows the rate of aging in laboratory rodents but the mechanism of action is unknown. DR is known to induce beneficial effects in a variety of tissues and organ systems. DR also maintains high levels of physical activity over the life span. We tested the hypothesis that lifelong physical activity is an important component of the anti-aging action of DR. Male specific pathogen-free Fischer 344 rats were divided into 4 groups at 6 weeks of age: A: fed old libitum; AE: fed ad libitum and in cages with running wheels; B: fed 60% ad libitum; BE: fed 60% ad libitum and in cages with running wheels. Running activity and spontaneous cage activity were measured over 24 hours and over the life span. Metabolic rate was measured indirectly by analysis of air entering and leaving cages. AE rats exhibited low levels of running activity and ran very little beyond 6 months of age. In contrast, BE rats sustained high running levels even after all A and AE rats had died. High levels of wheel running did not decrease spontaneous cage activity. Median life span (50% survival) was in the order A = AE < B < BE. Ten percent survival was in the order A = AE < B = BE. BE rats had greatest median life span and also highest specific metabolic rate. Exercise and DR altered pathology: At death BE rats had a high incidence of cardiomyopathy, whereas A and AE rats had high incidence of chronic nephropathy and pituitary tumors. The data indicate that increased physical activity is probably not an important factor in the action of DR on aging.

[A case of paraganglioma of urinary bladder in a 96-year-old female].

A 96-year-old woman was referred to our hospital with gross hematuria. Cystoscopy, computed tomography and magnetic resonance imaging revealed a submucosal bladder tumor. Incomplete transurethral resection was performed with no intraoperative complications. The histopathological diagnosis of nonmalignant paraganglioma was confirmed by immunohistochemical staining. This patient is the oldest of the 49 patients with paraganglioma of the urinary bladder reported in the Japanese literature.

VEGF and bFGF mRNA are expressed in ethylnitrosourea-induced experimental rat gliomas.

1. Which angiogenic growth factors actually mediate tumor growth in ethylnitrosourea (ENU)-induced gliomas in rats was examined. 2. In situ hybridization histochemistry with digoxigenin-labeled oligonucleotide probes was used to investigate the cellular expression and distribution of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) mRNAs in ENU-induced gliomas. 3. Both VEGF and bFGF mRNAs were not detected in normal gial cells but in ENU-induced glioma cells. 4. Our results suggest that the growth of ENU-induced glioma may be regulated by multiple angiogenic growth factors and that these gliomas may proliferate by synthesizing such growth factors.

Pleomorphic adenoma of the breast: report of a case.

We present herein the case of a 74-year-old woman found to have a pleomorphic adenoma, or so-called mixed tumor, of the breast. The patient presented with a hard mass, 3 cm in diameter, located just beneath the left areola. Physical examination suggested a diagnosis of breast carcinoma; however, the findings of both ultrasonography and mammography indicated a benign neoplasm. Aspiration biopsy cytology was evaluated as class III and the diagnosis of pleomorphic adenoma was finally confirmed by examination of an excisional biopsy specimen. As it is a rare benign tumor, pleomorphic adenoma of the breast has been mistaken for a malignant tumor clinically, mammographically, cytologically, and even by frozen section. Thus, an awareness of this disease in the breast will help to prevent misdiagnosis and overaggressive surgery.

Effects of lifelong dietary restriction on somatotropes: immunohistochemical and functional aspects.

We investigated the effects of lifelong dietary restriction (DR) on growth hormone (GH)-immunoreactivity and secretory function of somatotropes using a computerized image analysis in tissue sections and a reverse hemolytic plaque assay (RHPA) in dispersed pituitary cells of male F344 rats. DR did not prevent an aging-related reduction in GH immunoreactivity in somatotropes. However, it did augment the mean optical density of GH-immunoreactive area in somatotropes, although it did not alter the immunoreactive area in somatotropes, suggestive of a greater amount of immunoreactive GH in somatotropes of DR rats. DR also increased the percentage of aggregated GH-immunoreactive area in the anterior lobe, indicating a higher cell density of immunoreactive somatotropes. RHPA also confirmed the presence of larger proportions of GH-secreting cells in dispersed cells of DR rats in response to GH-releasing hormone (GHRH) at 24 months, although no change by DR was observed in the estimated amount of GH secreted from individual cells. Our results suggest that lifelong dietary restriction may influence the cytoplasmic GH-content at the steady state, while not modulating the responsiveness of individual somatotropes to GHRH for GH release, and that the major action of DR on somatotropes is an increment in the cell number.

Influence of dietary components on occurrence of and mortality due to neoplasms in male F344 rats.

The influence of dietary components on the occurrence of, and mortality from spontaneous neoplasms in male F344 rats was investigated. The dietary regimens studied included restriction of specific dietary components (energy, fat, protein and mineral), as well as different sources of dietary protein (casein, soy protein and lactalbumin). A statistical approach based on contributing causes of death was used to obtain the mortality due to all neoplasms, and the relative onset rate of frequently observed neoplasms, e.g., leukemia, pituitary adenoma, testicular interstitial cell tumor, etc. Only the regimen involving energy restriction reduced the mortality due to all neoplasms. Neither reduction of individual components without energy restriction, nor replacement of casein with soy protein or lactalbumin as the protein source affected mortality. Analyses of the relative onset rate of selected neoplasms also indicated that only a reduction of energy intake suppressed the occurrence of most of these neoplasms. Other dietary regimens, at most, suppressed a few types of neoplasm. It is concluded that a reduction in energy intake is a key dietary factor for the prevention of neoplastic diseases in rats.