Demonstration and diagnostic significance of pro-gastrin-releasing peptide in medullary thyroid carcinoma.

The expression of pro-gastrin-releasing peptide (proGRP) in medullary thyroid carcinoma (MTC) and other histological types of thyroid carcinoma was studied by an immunohistochemical technique, using polyclonal anti-proGRP antiserum. Immunoreactivity for proGRP was detected exclusively in MTC (n = 7); other histological types (n = 12) were all negative for proGRP. In addition, serum proGRP levels were elevated in patients with primary or recurrent MTC (n = 3), and they changed in parallel with serum calcitonin (CT), and carcinoembryonic antigen (CEA). These results show that proGRP, which is considered to be a specific marker for small-cell lung carcinoma, may be also a potential tumor marker for MTC, in addition to CT and CEA.

Chromosome rearrangement at 17q25 and xp11.2 in alveolar soft-part sarcoma: A case report and review of the literature.

BACKGROUND: Despite the characteristic histopathologic appearance of alveolar soft-part sarcoma (ASPS), its histogenesis remains unclear, and cytogenetic analysis of ASPS is limited to eight cases so far because of the extreme rarity of this disease. METHODS: The authors document a cytogenetic study of a primary case of ASPS in which a modern spectral karyotyping technique was used. RESULTS: A standard cytogenetic analysis of the primary tumor cells with G-banding revealed 46,XY, add(17)(q25) in 23 of 25 metaphases analyzed. This structural rearrangement of chromosome 17, involving band q25, was also present in 5 of 8 ASPS cases in the literature. Moreover, with the spectral karyotyping technique, the additional part of the long arm of chromosome 17 in the current case was found to originate from chromosome X, resulting in a final tumor karyotype of 46, XY, add(17)(q25).ish der(17)t(X;17) (p11.2;q25)(wcpX+). CONCLUSIONS: This case report documents a clonal chromosome abnormality of der(17)t(X;17)(p11.2;q25) in ASPS. The results of the current study indicate that further molecular analyses focused on 17q25 and Xp11.2 are of interest and could help to elucidate the pathogenesis of ASPS.

Expression of smooth muscle calponin in synovial sarcoma.

Purpose. Histogenesis of synovial sarcoma remains controversial and reliable molecular markers for diagnosis are necessary. Expression of basic calponin, a smooth muscle differentiation-specific actin-binding protein, was studied in synovial sarcoma.Subjects and Methods. The basic calponin gene and the gene product were analyzed by reverse transcription PCR analysis (RT-PCR) and immunohistochemistry in 14 synovial sarcomas and a human synovial sarcoma cell line (HS-SY-II).Results and Discussion. Immunoreactivity for basic calponin was detected in the cytoplasm of 6 synovial sarcomas (43% positive). In the basic calponin-positive tumors and the HS-SY-II cells, expression for smooth muscle-specific genes, including basic calponin and SM22alpha , was detected by RT-PCR, suggesting a lineage relationship between synovial sarcoma cells and smooth muscle-like mesenchymal cells.Conclusions. A subset of synovial sarcomas expressing the basic calponin gene and the gene product were identified. The basic calponin may have potential utility as a novel molecular marker identifying certain synovial sarcomas.

Expression of vascular endothelial growth factor in normal pituitary cells and pituitary adenomas producing adrenocorticotropic hormone.

Vascular endothelial growth factor (VEGF) induces endothelial cell proliferation and an increase in capillary permeability. Because the anterior pituitary gland and pituitary adenomas are highly vascular, expression of VEGF was examined immunohistochemically. Some normal pituitary cells stained positively for VEGF, and restaining for ACTH, prolactin, TSH, LH, FSH, and S-100 protein after VEGF staining revealed that almost all cells staining positively for ACTH also stained for VEGF. Only adenomas staining positively for ACTH stained for VEGF. These results suggest that VEGF is produced by normal pituitary cells and adenomas producing ACTH.

Enhanced efficacy of nilvadipine in hypertensives whose raised ambulatory blood pressure is sustained during sleep.

This study was designed to clarify the relationship between the antihypertensive effects of the calcium antagonist nilvadipine, and circadian changes in blood pressure. Based on measurements using an ambulatory blood pressure monitoring system (ABPM), 17 outpatients with untreated essential hypertension were divided into two groups: a sustained hypertensive group (with a fall in blood pressure during sleep < 10%, n = 7) and a waking time hypertensive group (with a fall in blood pressure during sleep > or = 10%, n = 10). During treatment with nilvadipine (8 mg/day, > or = 2 weeks), patients were reexamined by ABPM. The antihypertensive effect of nilvadipine was significantly and negatively correlated with the night time fall in blood pressure: this effect was significantly greater in the sustained hypertensive group than in the waking time hypertensive group. These data suggest that the long acting calcium antagonist nilvadipine has more potent antihypertensive effects in patients with sustained hypertension ("nondippers") than in those whose hypertension lessens during sleep ("dippers").

Antiangiogenic agent (TNP-470) inhibition of ectopic bone formation induced by bone morphogenetic protein-2.

Bone morphogenetic protein (BMP) is a potent inducer of ectopic bone formation, and TNP-470, a synthetic analog of fumagillin, is an antiangiogenic agent that strongly inhibits neovascular formation in vivo. We investigated the effects of TNP-470 on BMP-induced ectopic bone formation to clarify the role of angiogenesis in bone formation. Collagen pellets containing recombinant human BMP-2 (rhBMP-2) were implanted beneath the fasciae of dorsal muscles in mice. By daily subcutaneous administration of TNP-470, ectopic new bone formation was inhibited in a dose-dependent manner. Histological examination revealed that TNP-470 prevented proliferation of mesenchymal cells and chondrogenesis at the initial step of endochondral bone formation. Immunohistochemical staining with a specific antibody against bone morphogenetic protein type IA receptor showed that TNP-470 reduced the number of receptor-positive cells surrounding the BMP pellets. The inhibitory effect of TNP-470 on bone formation continued during the period of its administration, and discontinuation of treatment was followed by the resumption of the whole process of endochondral bone formation. This study showed that TNP-470 reversibly inhibits the biological activity of rhBMP-2 in the early stage of bone induction, suggesting that angiogenesis may play an essential role in the recruitment of BMP-receptor-positive cells that can respond to rhBMP-2 and differentiate into chondrocytes and/or osteoblasts.

Inhibition by ginsenoside Rg3 of bombesin-enhanced peritoneal metastasis of intestinal adenocarcinomas induced by azoxymethane in Wistar rats.

The effects of concomitant use of bombesin and ginsenoside Rg3 on the incidence of peritoneal metastasis of intestinal adenocarcinomas induced by azoxymethane were investigated in male inbred Wistar rats. From the start of the experiment, rats were given weekly s.c. injections of azoxymethane (7.4 mg/kg body weight) for 10 weeks and s.c. injection of bombesin (40 microg/kg body weight) every other day, and from week 20, s.c. injections of ginsenoside Rg3 (2.5 or 5.0 mg/kg body weight) every other day until the end of the experiment in week 45. Bombesin significantly increased the incidence of intestinal tumors and cancer metastasis to the peritoneum in week 45. It also significantly increased the labeling index of intestinal cancers. Although administration of a higher dose of ginsenoside Rg3 with bombesin had little or no effect on the enhancement of intestinal carcinogenesis by bombesin, the location, histologic type, depth of involvement, infiltrating growth pattern, labeling and apoptotic indices and tumor vascularity of intestinal cancers, it significantly decreased the incidence of cancer metastasis. These findings indicate that ginsenoside Rg3 inhibits cancer metastasis through activities that do not affect the growth or vascularity of intestinal cancers.

Dramatic response of postthymomectomy myasthenia gravis with multiple lung nodules to corticosteroids.

A 36-year-old woman underwent thymomectomy and left pneumonectomy with total pleurectomy for stage IVa invasive thymoma without myasthenia gravis. Six years later, a crisis of myasthenia gravis developed associated with multiple lung nodules. Steroid therapy induced remarkable regression of the lung nodules that has persisted for more than 11 months. Concurrently, the clinical severity of her myasthenia gravis markedly decreased and recovered to the extent of permitting her to return to normal life.

Prognostic significance of Ki67 (MIB1) proliferation index and p53 over-expression in chondrosarcomas.

To investigate the prognostic significance of the Ki67 (MIBI)-proliferation index and p53 over-expression in chondrosarcomas, we retrospectively analyzed a cohort of 29 patients with chondrosarcomas using immunohistochemical assays with MIBI and p53 monoclonal antibodies on formalin-fixed, paraffinembedded tissue samples with microwave preparation. We also assessed 19 patients with benign cartilaginous tumors as a control group. There was a significant positive correlation between MIBI index and tumor grade in chondrosarcomas, while there was no significant difference in the MIBI index between the grade-1 chondrosarcomas and the benign cartilaginous tumors. Patients categorized in the high-MIBI-index group had a significantly lower survival rate than those in the low-index group. Moreover, in analyzing the sub-set of the patients with grade-II chondrosarcomas, it was found that they could be prognostically sub-divided according to MIBI index. The p53 index also significantly correlated with patient survival, and there was significant correlation between the MIBI index and the p53 index. However, in multivariate analysis, only the MIBI index and tumor grade proved to be independent prognostic indicators of chondrosarcomas. These results demonstrate that the MIBI index can be a useful procedure for assessing tumor grade in chondrosarcomas, especially for determining the prognosis of patients with grade-II chondrosarcoma.

Distribution and subcellular localization of a growth inhibitory factor in hamster liver and its intracellular partner(s).

The subcellular, intralobular distributions and intracellular partner(s) of a factor which inhibits the proliferation of cell growth (Hashimoto C. et al. (1994) Biochim. Biophys. Acta 1221, 107-117) were determined in hamster livers, using a combination of immunological and biochemical techniques. The IgG fraction from an antiserum raised against the growth inhibitory factor with 37 kDa was shown to be highly specific for the antigen. The nuclear and cytosolic fractions demonstrated inhibitory effects on cell growth and Western blot analysis revealed that both fractions contained the immunoreactive 37 kDa protein with the anti-inhibitory factor IgG but microsomal and mitochondrial fractions did not. The nuclear and cytoplasmic localization of the inhibitory factor were further confirmed by immunochemical staining mediated through the immune IgG and an avidin-biotinylated horseradish peroxidase complex, the parenchymal liver cells were clearly stained, but endothelial and connective tissue cells were not. Although some staining was evident throughout the liver parenchyma, the hepatocytes with most intensively stained nuclei were located in the periportal region. In the liver from hamsters 6 days old or the regenerating hamster livers 3 days after partial hepatectomy, the staining intensity was low and the number of hepatocytes with the inhibitory factor positive nuclei was very few compared with the adult hamster livers. In primary cultures of the isolated hepatocytes from adult hamster the inhibitory factor disappeared from nuclei after incubation for 24-48 h. The extracts of hepatic nuclei from adult hamsters were immunoprecipitated with either the anti-growth inhibitory factor IgG or a monoclonal antibody to the RM protein. The growth inhibitory factor and the RB protein coprecipitated in each case, implying that the proteins were complexed with each other in the nuclei. The RB protein family is composed of two sets of species, an un- or underphosphorylated species and a hyperphosphorylated one. It was suggested that the factor bound preferentially to the un- or underphosphorylated member of the family.

Auditory threshold shift following meatal pressure change in ossicular disorders.

Auditory threshold shifts for air conduction following meatal pressure changes were examined in 30 normal ears and in 27 ears with ossicular abnormalities. Negative and positive changes in meatal pressure caused threshold elevation at frequencies lower than 1,000 Hz in all normal ears. In the ears with ossicular fixations, air pressures were less effective in attenuating hearing as compared with normal ears. Clearly different results were obtained in the patients with incudostapedial disconnection without stapes fixation. Thresholds were elevated by positive pressure, although marked threshold gains were measured by negative pressure at low frequency tones. Differences in threshold levels between the air pressures of +200 and -200 mm H2O were 26 to 40 dB at 250 Hz. This marked reverse effect in direction of the threshold shifts cannot be explained only by relative compliance changes in the tympanic membrane. Possible mechanisms underlying this phenomenon are discussed.

Elevated calcium level induces calcium-dependent proteolysis of A-CAM (N-cadherin) in heart--analysis by detergent-treated model.

Calcium overload induces cardiac muscle cell dysfunction in cardiovascular diseases. We investigated the effects of elevated calcium level on adherens-junction-specific cell adhesion molecule (A-CAM). Incubation of Triton X-100-treated canine heart homogenate in the presence of Ca2+ reduced the content of A-CAM. Reduction in A-CAM requires milli-molar Ca2+ and was inhibited by protease inhibitors, leupeptin and calpeptin. Immunohistochemical observation revealed that m-calcium-activated neutral protease (m-CANP) was colocalized with A-CAM in intercalated disks. These data suggested that m-CANP proteolyzes A-CAM in response to calcium overload in cardiac muscle.

Disruption of cell-cell adhesion in an inbred strain of hereditary cardiomyopathic hamster (Bio 14.6).

OBJECTIVE: Disarrangement of cardiomyocytes is a pathological characteristic of dilated cardiomyopathy. Hereditary cardiomyopathic hamster Bio 14.6, a model of dilated cardiomyopathy, displays disorder of cardiomyocyte arrangement. The aim of this study was to analyse the disturbance of cell alignment from the point of view of the cell-cell adhesion system in Bio 14.6. METHOD: Cardiomyopathic hamster Bio 14.6 was used as a model of dilated cardiomyopathy. Histological study was performed by light and electron microscopy. Disorder of the adherens junction-specific cell-adhesion molecule (A-CAM) was analysed by immunofluorescent microscopy and immunoblotting with anti-A-CAM antibody. RESULTS: Hematoxylin-eosin staining revealed that intercalated disks were identifiable less clearly in cardiomyopathy than in a normal cardiac muscle. It was disclosed by electron microscopy that cardiomyocytes adhered to each other with reduction in subsarcolemmal electron density at intercalated disks in Bio 14.6 compared with normal hamsters. We examined the localization of the A-CAM molecule in heart by immunofluorescent microscopy. In contrast to normal cardiac samples, fluorescence was weak in intensity and unclearly demarcated in the Bio 14.6 hamsters. We measured the content of A-CAM in the heart. In Bio 14.6 hamsters, the content of A-CAM was 60 +/- 11% of that measured in normal adult hamsters. A-CAM was reduced to a lesser extent (81 +/- 12%) in the newborn hamsters. CONCLUSIONS: In Bio 14.6 hamster, structural disturbance of the intercalated disks was found on histological examination of the heart. Biochemically, A-CAM, which plays a role in intercellular adhesion in intercalated disk areas, decreased significantly. These results suggest that cardiomyopathy may be accompanied by structural disruption of cell-cell adhesion in intercalated disk regions, which may lead to the pathological feature of disarranged cardiomyocytes.

Auditory brainstem responses in leptomeningeal metastatic spread. Presentation of 2 cases.

Auditory brainstem responses (ABRs) were recorded in 2 patients with hearing loss caused by leptomeningeal metastatic spread. These recordings showed similar characteristic findings. The absolute latencies of wave V and interwave latencies I-V were exceedingly increased. Definite effects on the wave morphology and latency of wave V were observed with the use of high repetition rates. It is surmised that the hearing loss caused by leptomeningeal metastatic spread is mainly an effect on the auditory nerve and/or the cochlear nucleus. ABR examination is of clinical value in detecting functional abnormalities resulting from leptomeningeal metastatic spread.

[Ventilation and passages in the middle ear. A study by HRCT in patients with attic cholesteatoma].

Operative records of 75 patients with acquired attic cholesteatoma were evaluated and compared with preoperative HRCT findings. The cholesteatoma extensions were classified into five groups as follows; Group 1 (cholesteatoma limited to the attic, 9 cases), Group 2 (cholesteatoma occupying both the attic and the aditus, 5 cases), Group 3 (cholesteatoma extending down to the posterior tympanum, also occupying an area as in Group 2, 6 cases), Group 4 (cholesteatoma occupying the attic, the aditus and the mastoid antrum, 14 cases), and Group 5 (cholesteatoma extending down to the posterior tympanum, also occupying an area as in Group 4, 41 cases). Ventilatory conditions, or the existence of soft tissue density, were evaluated by HRCT at such locations as the supratubal recess, mesotympanum, anterior and posterior parts of the tympanic isthmus, epitympanum, and mastoid antrum. Results are as follows. In Group 1, all these locations were aerated with the exception of a few cases. In Group 2, complete opacification was observed in almost all of the antrums. Soft tissue masses involved the anterior and/or posterior parts of the tympanic isthmus in both Groups 3 and 5. In Group 3, the posterior part of tympanic isthmus was less aerated than the anterior part of tympanic isthmus. In Group 5, all locations were filled with soft tissue density except the supratubal recess and the mesotympanum, where some degree of aeration was observed. These results indicate that blockage of the ventilatory passages is not essential for formation of an attic cholesteatoma. Soft tissue density in HRCT is not the cause, but rather, the result of extension of a cholesteatoma.

[A case of hypopharyngeal stenosis caused by late radiation injuries--reconstruction of the hypopharynx with myomucosal tongue flap].

The authors reported a 47-year-old man with hypopharyngeal stenosis caused by late radiation injuries. At the age of ten he underwent irradiation (3000rads) to the neck because of the cervical lymphadenopathy. He had keroid skin change at the age of 19, hypothyroidism since 26, right cervical and brachial plexus neuropathy since 33, and paralysis and papilloma of right vocal cord at 34. And at the age of 41 he underwent tracheostomy owing to laryngeal stenosis. In November 1984 (at age 43) he felt abnormal sensation on the throat but had no dysphagia nor misdiglutition. On November 1987 he had difficulties of swallowing, and could not take anything but fluid. At that time he was diagnosed as hypopharyngeal stenosis. With steroids and antibiotics his difficulties of swallowing were reduced. He experienced the same difficulties on April 1988. Since December 1988 his dysphagia got worse and was not recovered with medication. On May 17 1989, laryngopharyngectomy was performed. At the level of cricoid cartilage hypopharynx was resected. As for the posterior wall, pharynx and cervical esophagus were fixed to prevertebral fascia and anastomosed with end-to-end. And antero-lateral defects were reconstructed with myomucosal tongue flap. Postoperatively he could eat orally. On the basis of the experience of this case and the review of the literature the authors conclude that myomucosal tongue flap is one of alternatives for hypopharyngeal reconstruction.

[The role of high-resolution CT in evaluating disease of the posterior tympanum].

The posterior tympanum consists of several irregular eminences, ridges, and sinuses. These sinuses do not communicate with the mastoid air cell system, and cholesteatoma and/or granulation tissue prefer to fill them. This area cannot be visualized with ease by the usual surgical approach, and is thus quite important for surgeons. Recent developments in high-resolution computed tomography (HRCT) allow more than ever precise identification of subtle changes in the posterior tympanum. Axial HRCT sections provide essential information for the preoperative evaluation of the posterior tympanum. The performance of surgeons has been promoted both in evaluation and treatment planning of patient with suspected posterior tympanic lesions. HRCT findings were compared with the operative findings in 52 patients (54 ears) who had the operation for chronic otitis media. Important findings by HRCT diagnosis of posterior tympanum are as follows: (1) Bone destruction in the posterosuperior part of the tympanic anulus and in the lateral wall of the facial sinus are the important findings for the diagnosis of invasion of cholesteatoma in the posterior tympanum. (2) Similarly important finding is the bone destruction of the pyramidal eminence. This was found in 6 of 9 cases with cholesteatomas extending into the sinus tympani. (3) The soft tissue density in the posterior tympanum does not necessarily indicate pathological processes. Effusion in the posterior tympanum is imaged as soft tissue density, and can not be differentiated from cholesteatoma or granulation by present HRCT.

[The use of intraoperative auditory evoked brainstem responses in the treatment of congenital ossicular anomalies].

Intraoperative auditory brainstem responses (ABRs) were monitored in 3 patients undergoing middle ear surgery for congenital ossicular anomalies. It was investigated whether this technique was capable of providing useful information to the operating surgeon. After the induction of anaesthesia and intubation, standard disc electrodes were attached to the vertex (active), ipsilateral mastoid (reference), and forehead (ground). These electrodes were draped out of the surgical field. The general anaesthesia was maintained by inhalation of enflurane. Stimulus intensity was decreased from 80 dBnHL in 10 dB steps until no response was obtained. Thus latency-intensity functions for wave V of ABR were obtained. First (A) recording was performed before the incision. The tympanomeatal flap was then elevated and the bony annulus overloading the oval window was drilled out. This enabled to visualize and confirm the anomalies of the ossicles. Before reconstructing the ossicles, the tympanomeatal flap was returned to its normal position and second (B) recording was obtained. During the reconstruction, recordings (C1, C2,...) were repeated as indicated. Then it was confirmed whether the effective reconstruction was obtained for auditory function. The results suggest that latency of wave V was significantly decreased when the effective reconstruction of the ossicular chain was done. Latency-intensity function is shifted to the left of A and B functions. Intraoperative auditory assessment using ABR in the reconstructive surgery for the ossicular channel is promising in providing useful information for auditory function and in predicting postoperative hearing improvement.

[High-resolution computed tomography of posterior tympanum].

A complex set of sinuses, eminences and ridges lies in the posterior border of the tympanic cavity (posterior tympanum). The facial nerve canal is located between the facial sinus and the sinus tympani. The posterior limit is the posterior semicircular canal. The posterior tympanum is often the site of residual collection of granulation tissue or cholesteatoma, and is not directly visualized by the usual surgical approaches. Thus preoperative knowledge of cholesteatoma in these area is of obvious importance. It has been accepted that high-resolution computed tomography (HRCT) has the significant advantage to provide specific information of the middle ear. Contribution of the HRCT for diagnosis of posterior tympanum was examined by three temporal bone specimens and one hundred subjects with normal middle ear. In the experiments using the temporal bone specimens, each structure in the posterior tympanum was labeled by a fine needle. HRCTs were subsequently obtained to confirm the labeled structure. In one hundred subjects, it was examined whether each structure was also confirmed by the routine HRCT. Results are as follows: (1) Axial HRCT sections provided essential informations for the preoperative evaluation of the posterior tympanum. (2) Such bony structures as the pyramidal eminence and the pyramidal ridge were the most prominent structures on the posterior wall. (3) Laterally close to the pyramidal eminence lies one sinus which was marked by the needle placed in the facial sinus proper or in the lateral tympanic sinus; It is the facial sinus in a wide sense.(ABSTRACT TRUNCATED AT 250 WORDS)

An electrophysiologic study of experimental perilymphatic fistula.

This study was undertaken to elucidate the mechanism that causes sensorineural hearing loss in clinical cases with perilymphatic fistula. Perilymph was experimentally aspirated through the round window membrane in 17 guinea pigs. The extent of cochlear damage was examined electrophysiologically as well as histopathologically. Immediately after aspiration, several types of changes in summating potential (SP) were observed. Two animals without a polarity change of the SP showed only slight threshold changes in both cochlear microphonic and action potentials, and no specific histopathologic changes in the cochlea. Reversed polarity of the SP was observed in three animals, of which one showed a high-amplitude negative SP followed by rapidly progressive hearing loss. Bulging of Reissner's membrane was confirmed histopathologically in this case. The SP disappeared in the remaining 12 animals. In animals with profound electrophysiologic changes, bulging or rupture of Reissner's membrane and damaged hair cells were observed. These findings suggest that an abrupt change in perilymphatic pressure produces morphologic changes in the membranous labyrinth, causing changes in the vibration function of the cochlear partition and in the function of the organ of Corti. Abrupt pressure imbalance may be a causative factor of sensorineural hearing loss in the case of perilymphatic fistula.