Detection of point mutation for human growth hormone in patients with anti-pituitary antibody positive type 1 diabetes mellitus.

We investigated genetic mutations in the coding region of the human growth hormone (hGH) gene in anti-pituitary antibody (APA)-positive patients with type 1 diabetes mellitus (n = 6) or autoimmune thyroid diseases (n = 10) and in APA-negative, healthy controls (n = 10). A point mutation in the hGH gene was discovered in two patients with type 1 diabetes mellitus. No mutations were found in the hGH gene in control subjects, patients with autoimmune thyroid diseases (Hashimoto's thyroiditits, Graves' disease) or in the remaining four patients with type 1 diabetes mellitus. The mutation was located in the coding region for the second amino acid in the N-terminal region of hGH. This point mutation was identified in codon 2 in exon 2 of the hGH gene. We successfully developed an allele-specific amplification method for detecting this mutation using the polymerase chain reaction.

Anti-pituitary antibody-induced multiple endocrine disorders in mice.

Circulating anti-pituitary antibodies (APAs) have been detected in patients with autoimmune diseases, although the role of autoantibodies in the pathogenesis of autoimmune diseases is still unclear. With the aim of elucidating the autoimmune mechanisms involved in patients with multiple endocrine disorders, we evaluated the pathological changes in the pituitary gland, thyroid, pancreas and adrenal gland of mice, both wild-type and using a murine model of autoimmune thyroid disease [MRL-lpr/lpr] that had been immunized with murine, rat, porcine or human pituitary glands. In four of seven mice, a 22 kD band corresponding to APA was detected by Western blotting in the serum from mice that had been immunized with human pituitary tissues but not in the serum from mice immunized with rat or pig tissue. Inflammatory changes were detected in all groups of mice, occurring in the hypophysis, pancreas and adrenal glands but not in the thyroid. In conclusion, APA-induced autoimmune endocrine disorders are likely to be important for studying the mechanisms involved in autoimmune syndromes.

Reduction of viral myocarditis in mice lacking perforin.

Cellular immunity plays an important role and contributes to morbidity and mortality in the development of cardiomyopathy secondary to acute myocarditis. Lymphocytotoxicity has been proposed as due to infiltrated cytotoxic T lymphocytes (CTLs). Perforin is thought to be a major factor responsible for the cytolytic properties of the CTLs. To investigate whether mice lacking perforin have enhanced severity of viral myocarditis and acceleration of myocardial apoptosis, perforin knockout (KO) mice were infected with encephalomyocarditis virus. The average cardiac viral titer in perforin KO mice was not significantly different from that in the wild type mice. The heart weight/body weight ratio in perforin KO mice on days 4 and 12 were the same in the wild type mice (n=4 of each). Hearts in perforin KO mice were smaller than that in perforin + mice and grades of myocardial necrosis and lymphocyte infiltration were significantly reduced than in perforin + mice on day 12 after viral inoculation. In the perforin KO mice, the number of apoptotic cells was significantly higher than in wild type on day 4. Perforin blockade induced the early induction of myocardial apoptosis and suppressed the late onset of myocardial injury by cellular autoimmunity. Blockade of perforin gene expression at an early stage may limit viral myocarditis for therapeutic intervention.

Significant regional heterogeneity of coronary flow reserve in paediatric hypertrophic cardiomyopathy.

Previous studies have indicated that cardiac events in young patients with hypertrophic cardiomyopathy (HCM) are related to ischaemia rather than to arrhythmia. We measured coronary flow reserve in paediatric HCM and compared the values with those in adult HCM. We studied 12 patients with HCM including six paediatric (<20 years old; mean 13 years) and six adult patients (>20 years old: mean 62 years), and six healthy young adults (mean 29 years) as controls. Every patient underwent magnetic resonance imaging (MRI) for anatomical assessment. Myocardial blood flow at rest and after dipyridamole infusion was measured with dynamic nitrogen-13 ammonia positron emission tomography (PET). Partial volume effect was corrected for using the anatomical data obtained with MRI. In adult patients with HCM, coronary flow reserve in the hypertrophied septal region was not significantly different from that in the non-hypertrophied lateral wall (1.38+/-0.29 vs 1.77+/-0.39, respectively). In the paediatric patients, coronary flow reserve in the hypertrophied septal region was significantly lower than in the non-hypertrophied lateral wall (0.84+/-0.33 vs 2.74+/-0.90, respectively, P<0.01). In addition, coronary flow reserve in adult patients was lower than in control subjects both in the septal wall (1.38+/-0.29 vs 2.94+/-0.35, respectively, P<0.0001) and in the lateral wall (1.77+/-0.39 vs 2.85+/-0.69, respectively, P<0.05). In contrast, coronary flow reserve in paediatric patients was not significantly different from that in control subjects in the lateral wall (2.74+/-0.90 vs 2.85+/-0.69, respectively), while absolute reduction of myocardial blood flow was noted after pharmacological vasodilatation in the hypertrophied septal region. In conclusion, significant regional differences of coronary flow reserve were present in the paediatric patients with HCM. These results suggest that paediatric patients with HCM intrinsically have the potential to experience significant regional ischaemia even in the absence of coronary stenosis.

Non-invasive diagnosis of a Schwannoma by ultrasonography: a case study.

A 52-year-old female came to this institution complaining of a right leg mass lesion. Ultrasonography showed a 16 x 12 x 20mm hypoechoic solid mass lesion in the right leg. The patient underwent surgery during the diagnosis of Schwannoma. Preoperative diagnosis of Schwannoma is difficult by examination. Computerized tomography and magnetic resonance imaging, showed the origin of Schwannoma. However, the lesion was noted to be in a direct continuity with the cord-like echogenic structure consistent with a nerve by echography. We prefer to conduct preoperative examination by ultrasonography in patients with Schwannoma. In conclusion, we have reported a case of Schwannoma diagnosed by non-invasive ultrasonography.

Effects of PAMP on mRNAs coding for catecholamine-synthesizing enzymes in PC12 cells.

Proadrenomedullin N-terminal 20 peptide (PAMP) is a novel hypotensive peptide found in the N-terminal portion of the precursor of adrenomedullin (AM). Although PAMP and AM originate from the same precursor and exert both a potent hypotensive action, they seem to control blood pressure through different mechanisms. To gain new insight into the anticholinergic actions of PAMP, we determined the effects of PAMP on the tyrosine hydroxylase (TH)- and dopamine beta-hydroxylase (DBH) mRNA expression in the rat pheochromocytoma cell line PC12 stimulated by nicotine. PAMP (> or =1 microM) significantly inhibited the nicotine-induced increases of TH- and DBH mRNA expression in a concentration-dependent manner. Also, PAMP at the concentrations (> or =1 microM) significantly inhibited nicotine-induced cyclic adenosine monophosphate (cAMP) production. These results indicate that the anticholinergic hypotensive actions of PAMP can be explained, at least in part, by its inhibition of the expression of mRNAs coding for catecholamine-synthesizing enzymes, and that the inhibitory effect is mediated by the cAMP/protein kinase A pathway.

Use of technetium-99m sestamibi ECG-gated single-photon emission tomography for the evaluation of left ventricular function following coronary artery bypass graft: comparison with three-dimensional magnetic resonance imaging.

In patients who had undergone cardiac surgery (coronary artery bypass graft) and whose hearts showed abnormal movement during the cardiac cycle, we studied the accuracy of functional assessment using ECG-gated single-photon emission tomography (SPET) and the automated software developed by Germano et al. by comparing the findings with magnetic resonance (MR) images acquired three-dimensionally. Sixteen patients who had undergone cardiac surgery underwent 99mTc-sestamibi gated SPET (MIBI-g-SPET) and MRI on the same day. Left ventricular end-diastolic and end-systolic volumes (EDV, ESV) and ejection fraction (LVEF) were measured using MIBI-g-SPET and the aforementioned algorithm. Regional wall thickening was assessed using a four-point scale on MIBI-g-SPET and cine MRI. There was a good correlation between MIBI-g-SPET and MRI in respect of EDV (r=0.89), ESV (r=0.93) and LVEF (r=0.89). A high degree of agreement was found between the wall thickening scores obtained by MIBI-g-SPET and MRI in total segments (kappa=0.62) and in septal segments (kappa=0.67). It is concluded that ECG-gated perfusion SPET can provide regional and global functional information, including absolute volumes, in patients following cardiac surgery.

Leptin directly stimulates catecholamine secretion and synthesis in cultured porcine adrenal medullary chromaffin cells.

Leptin, a protein encoded by the ob gene, is an adipose tissue-derived signaling factor involved in body weight homeostasis. The hypothalamus is a major site of central action for leptin. However, mounting evidence indicates expression of leptin receptor mRNA in various peripheral organs including the adrenal medulla. Therefore, we investigated the effects of leptin on catecholamine secretion and synthesis in cultured porcine adrenal medullary chromaffin cells. We initially confirmed the expression of leptin receptor (Ob-Rb) mRNA in cultured porcine adrenal medullary cells. Murine recombinant leptin (>==50 nM) strongly induced the release of both epinephrine (E) and norepinephrine (NE) from chromaffin cells. Removal of external Ca(2+) significantly suppressed these effects. Also, leptin (>==1 nM) enhanced nicotine-induced increases in E- and NE. Leptin (1, 10, 100 nM) significantly increased tyrosine hydroxylase (TH) (a rate-limiting enzyme in the biosynthesis of catecholamine) mRNA levels in a concentration-dependent manner. Furthermore, leptin (1, 10, 100 nM) significantly induced increases in cAMP levels, suggesting that the stimulatory effects on TH mRNA are mediated, at least in part, by the cAMP/protein kinase A pathway. These results indicate that leptin directly stimulates catecholamine release and synthesis, which in turn may potentiate the anti-obesity effects of leptin.

Single breath-hold left ventricular volume measurement by 0.3-sec turbo fast low-angle shot MR imaging.

OBJECTIVE: In this study, we measured global cardiac parameters during a single breath-hold using T1-weighted single-shot turbo fast low-angle shot (turboFLASH) MR imaging. SUBJECTS AND METHODS: Ten cardiac patients who had undergone cardiac catheterization were examined with MR imaging to evaluate cardiac function. On average, 10 slices, which covered enough of the heart to measure the left ventricular volume, were acquired in a single breath-hold. During the acquisition of each slice, T1-weighted turboFLASH MR imaging with inversion recovery of both end diastole and end systole was sequentially accomplished twice in the same R-R interval. The end-diastolic volume, end-systolic volume, and ejection fraction calculated from MR images were compared with data obtained from left ventriculography. RESULTS: In these 10 cardiac patients, the ejection fraction calculated from turboFLASH MR images showed a good correlation with the measurements from radiographic left ventriculography (r = .87). The inter- and intraobserver variabilities in calculating the ejection fraction were 7.8% and 4.7%, respectively. CONCLUSION: Single-shot turboFLASH MR imaging with inversion recovery can provide data for the quantification of the ejection fraction during a single breath-hold.

Assessment of regional and global left ventricular function by reinjection T1-201 and rest Tc-99m sestamibi ECG-gated SPECT: comparison with three-dimensional magnetic resonance imaging.

OBJECTIVES: The purpose of this study was to test the ability of reinjection thallium-201 and rest technetium-99m sestamibi ECG (electrocardiographic)-gated SPECT (i.e., reinjection-g-SPECT [single-photon emission computed tomography] and MIBI-g-SPECT) to determine regional and global functional parameters. BACKGROUND: The ECG-gated perfusion SPECT was reported to provide accurate left ventricular ejection fraction (LVEF) using an automated algorithm. We hypothesized that other various functional data may be obtained using reinjection-g-SPECT and MIBI-g-SPECT. METHODS: Reinjection-g-SPECT, MIBI-g-SPECT, and three-dimensional magnetic resonance imaging (3DMRI) were conducted in 20 patients with coronary artery disease. Regional wall motion (RWM) and wall thickening (RWT) were analyzed using semiquantitative visual scoring by each g-SPECT and 3DMRI. The left ventricular end-systolic and end-diastolic volumes (EDV, ESV) and LVEF estimated by reinjection- and MIBI-g-SPECT were compared with the results of 3DMRI. RESULTS: A high degree of agreement in RWM and RWT assessment was observed between each g-SPECT and 3DMRI (kappa >.70, p < .001). The LVEF values by reinjection- and MIBI-g-SPECT correlated and agreed well with those by 3DMRI (reinjection: r = .92, SEE = 5.9%, SD of differences = 5.7%; sestamibi: r = .94, SEE = 4.4%, SD of differences = 5.1%). The same also pertained to EDV (reinjection: r = .85, SEE = 18.7 ml, SD of differences = 18.4 ml; sestamibi: r = .92, SEE = 13.1 ml, SD of differences = 13.0 ml) and ESV (reinjection: r = .94, SEE = 10.3 ml, SD of differences = 10.3 ml; sestamibi: r = .97, SEE = 6.7 ml [p < .05 vs. reinjection by F test], SD of differences = 6.6 ml [p < .05 vs. reinjection by F test]). CONCLUSIONS: Reinjection- and MIBI-g-SPECT provide clinically satisfactory various functional data. These functional data in combination with the perfusion information will improve diagnostic and prognostic accuracy without an increase in cost or the radiation dose to the patients.

Natural history of colorectal carcinoma: can the tumor volume doubling time be predicted by radiologic findings or immunohistochemical variables?

BACKGROUND AND OBJECTIVES: The factors influencing the growth rate of colorectal carcinoma have not been determined. The aim of this study was to clarify the relationship between the doubling time (DT), morphology, and proliferating cell nuclear antigen, Ki-67 and p53 immunohistochemistry in colorectal carcinoma. METHODS: Thirty-three patients (37 lesions) were studied retrospectively. The DT was calculated and correlated with the initial and final tumor size, morphologic shape, and immunohistochemical results. RESULTS: The DT ranged from 2.4 to 48.0 months (mean: 12.0 months). The mean DT of the early-stage carcinomas was significantly longer than that of the advanced carcinomas. In the latter group, both slowly growing and rapidly growing tumors were observed. The DT showed no correlation with the initial or final size and shape of the tumors on radiographs, or with the immunohistochemical results. CONCLUSIONS: Our data revealed that it is not possible to evaluate the growth rate of colorectal carcinomas based on their morphological shape, cellular proliferative activity, or tumor suppressor gene activity.

Identification and initial characterization of stathmin by the differential display method in nerve growth factor-treated PC12 cells.

The differential display of mRNA is a new strategy to identify genes that are differentially expressed under altered conditions. We applied this method to determine differential gene expression in the rat pheochromocytoma cell line during differentiation induced by nerve growth factor (NGF). Three different mRNA species were isolated, and their differential expression was confirmed by RT-PCR. One of the mRNA species was identified as stathmin, a 19 kDa cytosolic protein attracting increasing interest for its role in signal transduction. In the NGF-treated PC12 cells, the expression of stathmin mRNA increased in a time-dependent manner, as assessed by northern blot analysis and RT-PCR. We also assessed by northern blot analysis how the expression of stathmin mRNA was altered in human pheochromocytomas (n = 5) compared with that in normal adrenal medulla tissue (n = 5). The mRNA concentrations were found to be significantly greater in the pheochromocytomas than in the normal tissues. It has been shown that stathmin mRNA concentrations are increased in various tumor cells. As pheochromocytomas are well-differentiated tumors of neural origin, it is not unexpected that stathmin mRNA is overexpressed in these tumors. Stathmin was isolated and identified as a differentially expressed gene by the differential display method in PC12 cells during differentiation induced by NGF. In addition, stathmin mRNA was found to be overexpressed in human pheochromocytomas. The mechanisms responsible for the up-regulation of stathmin mRNA during differentiation of PC12 cells and the significance of its overexpression in human pheochromocytomas remain to be determined.

Gastrointestinal submucosal tumors: evaluation with endoscopic US.

PURPOSE: To describe the endoscopic ultrasound (US) features of benign versus malignant submucosal tumors throughout the gastrointestinal tract. MATERIALS AND METHODS: One hundred nine patients aged 24-81 years suspected to have submucosal tumors (11 esophageal, 41 stomach, 24 duodenal, and 33 colorectal tumors) at barium studies or endoscopy underwent endoscopic US. The layer of origin, internal echo pattern, and lesion margin were analyzed by means of consensus and independent interpretation by three radiologists. RESULTS: Endoscopic US findings revealed several distinct patterns among various submucosal tumors. Sixteen (94%) of the 17 homogeneous lesions with histopathologic findings of malignancy were hypoechoic, although 29 (43%) of the 68 homogeneous lesions with histopathologic findings of benignity were similarly hypoechoic. Homogeneous lesions that were anechoic, of intermediate echogenicity, or hyperechoic were almost exclusively benign (39 [98%] of 40). In contrast, 23 (96%) of the 24 malignant lesions were heterogeneous (n = 7) or homogeneously hypoechoic (n = 16). The sizes of benign and malignant lesions were significantly different (P < .05). There was no significant difference in the echo pattern (i.e., homogeneous versus heterogeneous), but there was a significant difference in the proportion of hypoechoic versus nonhypoechoic lesions (anechoic, hyperechoic, or of intermediate echogenicity; P < .001). CONCLUSION: The differential diagnosis of gastrointestinal submucosal tumors is assisted with endoscopic US.

Isolated Crohn's disease of the gastroduodenum: a case report.

We report a case of isolated Crohn's disease in the gastroduodenum. A 33-year-old man was hospitalized due to epigastralgia. Endoscopic and double-contrast radiographic studies revealed edematous gastroduodenal mucosa with a deep, undermining ulceration in the gastric antrum, which changed after a short interval into a typical "cobblestoned" appearance. A gastrectomy was performed due to antral narrowing, and Crohn's disease was histologically confirmed. Our objective was to reveal the change from a deep, undermining ulceration to a typical "cobblestoned" appearance using a double-contrast barium study.

Colonic submucosal tumors: a new classification based on radiologic characteristics.

OBJECTIVE: Despite advances in gastrointestinal radiologic techniques, findings from barium enema studies cannot be used reliably to predict the histologic nature of a colonic submucosal tumor. Therefore, we generated a new and comprehensive classification system for all colonic submucosal tumors based on a correlation between their radiologic characteristics and their histopathologic features. MATERIALS AND METHODS: Between 1978 and 1990, 89 patients had 95 colonic submucosal tumors diagnosed on the basis of barium enema studies and confirmed by biopsy or surgery. Of these, 39 patients (44%) were symptomatic and 50 patients (56%) were asymptomatic. Single lesions (82 cases) and multiple lesions (12 cases of lymphoma and one case of blue rubber bleb nevus) were observed. The 95 lesions included 21 carcinoids, 27 malignant lymphomas, 17 lipomas, 10 lymphangiomas, six leiomyosarcomas, five leiomyomas, and nine others. We analyzed the radiologic findings and classified these lesions according to their appearance. RESULTS: All tumors were classified on the basis of their morphologic appearance into five types: (1) wide-based sessile lesion with gradually sloping margin and smooth surface (17 cases); (2) wide-based sessile lesion, more polypoid than the first type, with a smooth surface, with or without a central depression (49 cases); (3) wide-based sessile lesion with lobulated surface (six cases); (4) pedunculated lesion with smooth or granular surface (17 cases); (5) unclassified, not any of the previously mentioned types, may be diffusely stenotic or aneurysmal (six cases). The most common finding was the wide-based sessile lesion with a smooth surface and a possible central depression; next most common was the wide-based sessile lesion with gradually sloping margin and smooth surface; next was the pedunculated lesion with a smooth or granular surface. Eighty-six percent of the lesions were single; 14% were multiple. Histologic confirmation revealed mainly lymphoma (29%) and carcinoid (22%), of which the most common finding was a wide-based sessile lesion with a smooth surface, with or without a central depression. CONCLUSION: This new classification based on radiologic characteristics covers all varieties of colonic submucosal tumors. It can serve as a guideline for predicting a specific diagnosis of a colonic submucosal tumor on the basis of radiologic findings.

Peripheral expansion of gamma delta T cell receptor-positive cells in a patient with Crohn's disease.

A 13 year old boy with Crohn's disease had a prominent expansion (over 40%) of gamma delta T cell receptor-positive cells (gamma delta T cells) in the peripheral blood. The gamma delta T cells were gradually decreased from 48 to 22% in 18 months. Anti-gamma delta T cell receptor-triggered cytotoxicity was evident at onset and after 1 year. The significance of peripheral expansion of gamma delta T cells in a patient with Crohn's disease is discussed.

Superficial flat-type early carcinoma of the stomach.

A clinicopathologic study was done on 21 cases of superficial flat-type early gastric carcinoma (IIb type EGC). In one case there was the two IIb type EGC. Nine patients had no symptoms, whereas the other 12 had either epigastralgia, hematemesis, or anorexia. The preoperative diagnosis was accurate in 15 patients; eight were demonstrated by barium study, and 13 by endoscopy. The suspicious finding of IIb type EGC was either the disappearance or irregularity of the areae gastricae by barium study and a mucosal color change by endoscopy. Well-differentiated or moderately differentiated adenocarcinomas showed a slight redness of the affected mucosa whereas the poorly differentiated adenocarcinomas were pale in color. Histologically, many well-differentiated or moderately differentiated adenocarcinomas occupied the entire thickness of the mucosal layer whereas most of the poorly differentiated adenocarcinomas spread horizontally with preservation of non-cancerous glands and foveolae. The growth pattern was super type in ten lesions and small mucosal type in 12 and no pen-type growth was seen. Concerning the cell nuclear DNA ploidy pattern, 21 showed a low ploidy pattern and only one had a high ploidy pattern. The IIb type EGC seemed to have a less malignant potential from the viewpoint of growth pattern and DNA ploidy pattern. Care must be taken at the proximal line of excision of the tumor so as not to leave behind residual carcinoma cells.

Colorectal carcinomas incidentally detected in autopsied cases and inpatients.

During the past 20 yr, 17 colorectal carcinomas (0.47%) were incidentally detected among 3,638 autopsied patients without clinically evident colorectal carcinoma, including 2,232 males and 1,406 females, more than 40 yr old. Among the 15 male and two female index subjects, six (0.33%) were detected in the first and 11 (0.60%) in the second decade. During their survival periods, fecal occult blood studies were performed in 14 cases and positive in 12 (86%); however, two of them had gastric ulcers which were responsible for the occult blood. During the recent 11 yr, six cases (0.48%) of colorectal carcinoma (four of them males; two, females) also were detected among 1,249 inpatients who were examined by barium enema and/or colonoscopy, including 816 males and 433 females, 40 yr old, or more, in the Department of Radiology. Fecal occult blood was detected in four cases (67%) before colonic investigation. Compared with 708 surgically resected carcinomas, the incidental lesions from both sources were smaller, consisted of higher percentages of Dukes' A type, and arose predominantly from the sigmoid colon and, rarely, from the rectum. These results indicate that the prevalence of colorectal carcinoma and its predominance in the sigmoid colon have not only apparently but actually increased in Japan, apart from improved diagnostic capabilities, and that false-negative rates with occult blood tests were surprisingly low in these autopsied cases and inpatients.