Production and partial purification of proteases from Aspergillus oryzae grown in a medium based on whey protein as an exclusive nitrogen source.

Several attempts have been made to incorporate whey proteins into curd to increase cheese yield. For some types of cheese, degradation of whey proteins that have been incorporated into the curd would be required to obtain acceptable flavor and texture. On the basis of the high potential for protease synthesis in Aspergillus oryzae, sodium nitrate as a nitrogen source in a minimal medium for fungi, known as Czapek-Dox medium, was replaced with whey protein isolate to induce the protease to hydrolyze whey protein using A. oryzae AHU7146. A solid-phase medium adjusted to pH 6 was suitable for this purpose when incubation was carried out at 25°C for 2 wk. The application of column chromatography enabled the resolution of 3 proteolytic components (1, 2, and 3). With respect to optimal temperature and zymographic analysis, component 1 was similar to component 3. In contrast, component 2 was less abundant than the other components and exhibited activity in the alkaline pH region. The degradation of ß-lactoglobulin and α-lactalbumin in whey protein isolate solution by the crude enzyme was primarily attributed to the action of components 1 and 3, based on HPLC analysis and the N-terminal amino acid sequences; however, zymography demonstrated evident proteolysis due to component 2. Because heat-denatured whey protein aggregates were digestible by the crude enzyme, the proteolytic system from A. oryzae has the potential as an additive to stimulate the ripening of cheese enriched with whey protein.

Augmentation ileocystoplasty in patients with neurogenic bladder due to spinal cord injury or spina bifida.

STUDY DESIGN: Retrospective analysis of augmentation ileocystoplasty for neurogenic bladder. OBJECTIVES: To analyze the effects and complications of augmentation ileocystoplasty in patients with neurogenic bladder due to spinal cord injury (SCI) or spina bifida retrospectively. SETTING: Department of Urology, Kanagawa Rehabilitation Hospital, Japan. METHODS: We have treated 11 patients with neurogenic bladder due to SCI and 10 patients with spinal bifida with augmentation ileocystoplasty since 1989. The purpose of the treatment was to stop vesicoureteral reflux (VUR) and/or amelioration of urinary incontinence. In 17 of 21 cases, the antireflux operation was received simultaneously. All cases performed clean intermittent self catheterization postoperatively. RESULTS: Urinary incontinence improved in all cases and only transient recurrence of VUR was observed in the follow-up term. Complications occurred in patients with SCI, but they could be treated conservatively. CONCLUSION: Augmentation ileocystoplasty is a good treatment option for contracted bladder or VUR, which occurs in patients with neurogenic bladder.

[Development of new hydrophilic catheter kit for self-catheterization: study in pediatric cases].

PURPOSE: To increase the management of self-catheterization in children of school age, a catheter kit consisting of hydrophilic catheter and a packet containing sterilized water was developed. We evaluated the lubricating characteristic and clinical efficacy of this new catheter kit. MATERIALS AND METHODS: The catheter kit used in the study was a pocket-size plastic container in which a polyurethane catheter coated with hydrophilic polymer and a packet containing sterilized water were packed in combination. The lubricating characteristic of catheter was assessed by the measurement of friction value. For clinical assessment, male children aged over 6 years old who were doing self-catheterization at 17 medical institutions nationwide were selected as the subjects. The 32 children who had given informed consent (mean age: 11.6 years old) were asked to use the catheter kit continuously for 1 week. The results were investigated by a questionnaire survey in which the assessment before and after the use was expressed in scores. At the same time, urinalysis and urine culture were examined. RESULTS: The friction value of hydrophilic catheter was equivalent to or less than that observed by applying a lubricant to the conventional catheter. The comparison of conventional catheter with the kit indicated significantly higher scores (assessment in 5 grades expressed in scores) for the portability and operability of the kit. Though there was no significant difference in the ease of insertion between the two catheters, there were several comments that the kit got stuck in the urethra when it was withdrawn. The global assessment gave a significantly higher score to the kit and 30 (94%) of the 32 children wanted to use the kit continuously. No increase in hematuria which caused a clinical problem or no new apparent urinary tract infection occurred after the use of the kit. CONCLUSIONS: Compared with the conventional catheter, the hydrophilic catheter kit highly satisfied a large number of children at the time of self-catheterization. Depending on the condition of children, the kit is considered useful for continued self-catheterization for a long term.

Single nucleotide polymorphisms in an STS region linked to the Ncc-tmp1A locus are informative for characterizing the differentiation of chromosome 1A in wheat.

Homoeoalleles of Ncc confer nucleus-cytoplasm (NC) compatibility on NC hybrids of wheat with the D plasmon of Aegilops squarrosa. To dissect the chromosomal region containing Ncc, a RAPD marker linked to the Ncc-tmplA locus, which is located on chromosome 1A of T timopheevi, was sequenced and converted to a PCR-based sequence-tagged-site (STS) marker. Five single nucleotide polymorphisms (SNPs) between T timopheevi and T turgidum. were detected in a 509-bp genomic DNA fragment. Based on the SNPs, the STS alleles in 164 accessions from emmer wheat, timopheevi wheat and two einkorn wheats, T. urartu and T. boeoticum were surveyed by PCR-RFLP analysis. The sequence comparisons and PCR-RFLP analyses revealed nine alleles based on six SNPs. These SNPs were highly conserved within each group of wheat, and all groups could be distinguished by particular combinations of the SNPs. All accessions of T. urartu had one unique STS allele as compared with the others. Our results indicate that the SNPs in the STS marker linked to the Ncc-tmplA locus would be informative for studies of the differentiation of chromosome 1A in wheat.

Long-term analysis of suprapubic cystostomy drainage in patients with neurogenic bladder.

OBJECTIVE: We assessed the roles of suprapubic cystostomy in patients with neurogenic bladder and analyzed the complications and their courses. PATIENTS AND METHODS: We reviewed 118 patients with neurogenic bladder managed with suprapubic cystostomy. The original diseases were spinal cord injury in 90, degenerative disease of the central nervous system in 15, spina bifida in 6, cerebral palsy in 3, pontine bleeding in 1, Parkinson's disease in 1, brain tumor in 1, and dysgenesis of the external sphincter in 1. Fifty-six (62.2%) of spinal cord-injured patients demonstrated cervical damage. Renal function, urinary pH and white blood cell values were measured and evaluated after insertion. The stone-free rate after insertion was estimated by the Kaplan-Meier method. RESULTS: Indications for cystostomy were failure of clean intermittent catheterization in 62 (52.5%) and Credé's maneuver in 2, severe urethral damage in 30 (25.4%), replacement of urethral catheter in 3, worsening of the original disease in 15 (12.7%), deterioration of the general condition in 2, mental retardation in 2, and traumatic vesical rupture in 1. Frequent complications were formation of the bladder calculi in 30 (25%) and urinary leakage through the urethra in 11 (10%). No fatal complications occurred. The stone-free rates 5 and 10 years after insertion were 77 and 64%, respectively. The urinary pH of the stone-forming group was significantly higher than that of the stone-free group. The high urinary pH group (>7.24) had a higher risk of stone formation. CONCLUSIONS: Although continuous cystostomy drainage is not considered to be ideal management for bladder emptying, some patients with neurogenic bladder may benefit from this procedure.

Primary gastric Hodgkin's disease. Morphologic, immunohistochemical, and immunogenetic analyses.

Two cases of primary gastric Hodgkin's disease were examined morphologically and immunohistochemically. Immunogenetic studies were also made in one of the cases. Case 1 was diagnosed as nodular sclerosis and case 2 as mixed cellularity Hodgkin's disease. Large atypical mononuclear or multinucleated giant cells were immunohistochemically positive for BerH2, whereas they were negative for LCA, L26, DAKO CD3, MT1, UCHL-1, S100 protein, and MAC387. In case 1, large atypical cells examined immunohistochemically on frozen tissue specimens were positive for Ki-1 and negative for T- and B-cell markers. Southern blot hybridization studies performed on material from case 1 revealed germline configuration of immunoglobulin JH and T-cell-receptor beta-chain genes. To our knowledge, this is the first report in which primary gastric Hodgkin's disease has been studied immunogenetically. Although rare in incidence, the existence of Hodgkin's disease in the stomach must be noted.

Morphometric analysis of colorectal dysplasia with image processing.

Nuclear and structural abnormalities on the histological specimens of colorectal dysplasias were quantified by using 10 parameters. From the measured values, the following histological findings were obtained: progression from mild to severe dysplasia was associated with an increase in nuclear size, and nuclear arrangement became more irregular. There was, however, no difference between the nuclear size and arrangement in severe dysplasia and those in carcinoma. The nuclei were spindle shaped in mild and moderate dysplasia. On the other hand, they were relatively round in severe dysplasia and carcinoma. Structural abnormalities tended to increase with the histological abnormalities. These results correspond well with the histological abnormalities by which the pathologists grade dysplasia and diagnose carcinoma subjectively. Therefore, these 10 parameters might be useful for quantifying histological characteristics of colorectal dysplasias.

Primary gastric Ki-1 positive anaplastic large cell lymphoma: a report of two cases.

Two cases with primary gastric Ki-1 positive anaplastic large cell lymphoma are presented. Morphologic features of both cases involved pleomorphism of the neoplastic cells, fibrosis and lymphatic infiltration. The neoplastic cells in both cases were positive for BerH2 (CD30), LCA(CD45), lysozyme and alpha-1-antitrypsin (alpha 1-AT). In additional case, the neoplastic cells were additionally positive for MAC387 and alpha 1-antichymotrypsin (alpha 1-ACT). The neoplastic cells in these cases were negative for L26(CD20), UCHL-1(CD45RO), DAKO CD3 and epithelial membrane antigen (EMA). According to the results of the phenotypic studies, the authors consider that the neoplastic cells have some of the features of histiocytes. Both patients at 2 and 8 years after surgery without chemotherapy are disease free. This lymphoma is well known to be frequently misdiagnosed as undifferentiated carcinoma. Although rare in occurrence, recognition of this primary lymphoma in the stomach has a significant clinical implication, as the authors consider that its prognosis might be better than undifferentiated carcinoma of the stomach.

Primary gastric plasmacytoma: a morphological and immunohistochemical study of five cases.

Five cases of primary gastric plasmacytoma were studied histopathologically and immunohistochemically. Plasmacytoid cells proliferated diffusely in the propria mucosa, almost preserving the structure of gastric glands. Occasionally, intranuclear inclusions, giant cells, and needle-shaped crystalline inclusions were observed. The neoplastic nature could be suspected on the basis of these histological findings. Immunohistochemically, three cases were positive for IgM and two for IgA. IgM positivity was more commonly observed in the gastric plasmacytoma than in multiple myeloma. Another immunohistochemical study demonstrated that LN-1 negativity and anti-EMA antibody positivity might be an indicator to differentiate gastric plasmacytoma from other types of gastric lymphoma. Four cases of early-stage gastric plasmacytoma have been followed for 5-12 yr. No recurrence has been observed so far. These cases suggest that gastric plasmacytoma has a relatively good prognosis.

Morphometrical analysis of nuclear abnormality of tubular tumors of the stomach with image processing.

Nuclear size and shape on histological specimens were used to quantify nuclear abnormality in gastric tubular tumors. A valid technique for this morphometrical analysis by image processing was developed. Furthermore, the present study examined the nuclear size and shape to determine if they were reliable criteria to be used for differential diagnosis. Nuclear area, maximum diameter and maximum width on the histological specimens were defined as factors to represent nuclear size. Nuclear aspect ratio represented nuclear shape. Histological features revealed varied degrees of condensation of nuclei; this was due to different protocols for preparing the histological specimens. This finding indicated that the size of intermediate lymphocytes on the same specimens should be measured as a control in order to estimate the degree of nuclear condensation. The present study also demonstrated that at least 200 nuclei should be measured on the histological specimens to obtain accurate nuclear size and shape. Histologically, the nuclei in benign tubular adenoma and borderline lesions tended to be spindle-shaped, while those in well-differentiated type tubular adenocarcinoma tended to be round and swollen. The nuclei in borderline lesions were larger than those in benign tubular adenoma and smaller than those in well-differentiated type tubular adenocarcinoma. Our results for the above 4 factors corresponded well with these histological findings. The nuclear size and shape on histological specimens were concluded to be reliable criteria for assessing nuclear abnormality in gastric tubular tumors and useful ones for differential diagnosis.

Colliding malignant lymphoma and adenocarcinoma of the stomach: case report.

We report a case of malignant lymphoma presenting as the superior vena cava syndrome. This case is very interesting since both adenocarcinoma mucocellulare scirrhosum, Borrman type IV, and lymphoma were present in the stomach, forming colliding neoplasms. Rather than systemically disseminated, the lymphoma cells were localized in the stomach, the intestine, a few perigastric lymph nodes and in a mediastinal mass, which may have constituted a neoplastic thymus invaded by lymphoma cells from the stomach. The unique occurrence of the histological combination as gastric colliding neoplasms is discussed.

Assessment of new macroscopical classification of gastric malignant lymphoma.

We macroscopically classified gastric malignant lymphoma into ST type (submucosal tumor type), non-ST type (non-submucosal tumor type) and Combined type (combination of ST and non-ST type) and assessed this new classification by examining 60 surgically resected cases. Of these, 18 cases (21 lesions) were classified into ST type, 38 cases (54 lesions) into non-ST type and 4 cases (4 lesions) into Combined type. All the large cell, immunoblastic type cases were of ST type and most of the diffuse, small cleaved cell type cases and all the plasmacytoma cases were of non-ST type. Diffuse, large cell type and diffuse, mixed type cases did not seem to be biased toward any of these three macroscopical types. The non-ST type cases frequently showed an infiltrative growth pattern and reactive lymphoid cell hyperplasia adjoining the lesions, while the ST type cases frequently showed an expansive growth pattern without reactive lymphoid cell hyperplasia. The Combined type cases showed both infiltrative and expansive growth patterns in the same lesion. The macroscopical classification defined in this study can be considered useful in determining a range for surgical resection of gastric malignant lymphoma.

Morphometrical analysis of structural abnormality of tubular tumors of the stomach with image processing.

Four indices, index of tubular density, index of dispersion of tubular size, degree of complexity of tubular shape and frequency of complex-shaped tubuli, were defined to quantify the structural abnormalities of gastric tumors in morphometrical analysis by image processing. The values of each index corresponded well with the degree of each structural abnormality found pathologically. These indices were considered to be valid for representing their respective structural factors. There were significant differences among the mean values of the scores calculated by a formula using all 4 indices of benign tubular adenoma, borderline lesion and well-differentiated type tubular adenocarcinoma (P less than 0.05). Therefore, the discriminant formula was considered to be valid for integrating these indices and for representing structural abnormality of gastric tubular tumors.

Primary gastric mantle zone lymphoma. A report of two cases.

We report two cases of primary gastric mantle zone lymphoma. Histologic examination revealed numerous lymphoid vague nodules in the mucosa and submucosa of the resected stomach. The neoplastic cells in these nodules were slightly larger than small lymphocytes and had more or less cleaved nuclei. Immunostaining on paraffin-embedded sections showed that the neoplastic cells in the nodules of these cases were LN-1- and LN-2+ and had monotypic immunoglobulin (IgM-lambda and IgM-kappa, respectively). Immunostaining on frozen tissue specimens showed that the neoplastic cells or nodules were positive for surface IgM, surface IgD, alkaline phosphatase, and DRC-1. One third to two thirds of the cells were Leu-1+ (CD5+). These results indicated that these cases were primary gastric mantle zone lymphoma. More attention should be paid to primary gastric mantle zone lymphoma because this disease might be erroneously diagnosed as either pseudolymphoma or reactive lymphoid hyperplasia of the stomach.

Objective quantification of grade of atypia in epithelial tumors of the stomach by image processing.

An image processing technique to extract tubular and nuclear areas from histological specimens stained with hematoxylin was established. Two indices representing tubular density and nucleus-to-cytoplasm ratio of glands were examined in intestinal-type tumor of the stomach. In specimens clearly stained with hematoxylin, significant differences of brightness among nuclei, cytoplasm and stroma can be obtained. Image processing by computer enabled us to identify nuclei, cytoplasm and stroma by utilizing the differences of brightness among them. Both indices tended to increase with severity of histological atypia and their values in the tubular adenoma group were significantly different from those in the tubular adenocarcinoma group (P less than 0.05). Therefore, these two indices could be regarded as valid and objective measures of the grade of histological atypia. Eighty-four percent of tubular adenoma and tubular adenocarcinoma cases were classified correctly with the discriminant formula and critical value calculated from the two indices on the basis of measurement of the images magnified by 40 times. There is, however, still a non-negligible overlap of discriminant scores between the tubular adenoma group and tubular adenocarcinoma group. More indices representing histological atypia will be required to allow objective differential diagnosis between malignant and benign lesions.

Primary gastric malignant lymphoma. A morphological and immunohistochemical study of 38 cases.

Thirty-eight cases of primary gastric malignant lymphoma were studied morphologically and immunohistochemically. The Working Formulation was used for classification of non-Hodgkin's lymphoma (1). The results indicated that 37 cases were non-Hodgkin's lymphoma while the remaining case was Hodgkin's disease. Thirty-three cases (89%) of non-Hodgkin's lymphoma were considered to be of B cell origin and 2 cases (5%) of histiocytic origin. No case was considered to be of T cell origin. We suggest that the majority of primary gastric malignant lymphomas are derived from follicular center cells, and that gastric plasmacytoma is not as rare as previously described.

Detection of anti-cerebral autoantibodies in schizophrenia and Alzheimer's disease.

Autoantibodies to cerebrum were investigated in the sera from schizophrenia, Alzheimer's disease and healthy subjects by indirect immunofluorescence method. Four patterns of staining were observed: type A (neuronal cell), type B (blood vessel), type C (nucleus) and type D (glia and miscellaneous cell). Among rat, guinea pig and rabbit cerebrum, rabbit cerebrum proved best (low non-specific binding and high sensitivity) for the detection of circulating autoantibodies to cerebrum. With schizophrenia, 13 out of 46 cases (28.3%) were positive for autoantibodies to neuronal cells and 6/13 of those were not absorbed by liver acetone powder. With Alzheimer's disease, 25/63 (39.7%) were positive for autoantibodies to neuronal cells and 6/25 were not absorbed by liver acetone powder.

[An autopsy case of esophageal cancer with brain metastasis].

A rare case of esophageal cancer with brain metastasis is reported. A 54-year-old man complaining of dysphagia was admitted to our hospital. Endoscopic and radiographic examinations revealed an advanced esophageal cancer with supraclavicular lymph node metastasis. Five months later, muscle weakness of right upper extremities developed. A brain CT scan revealed a low density area with a ring enhancement in the left front parietal region. The patient developed right hemiplegia and delirium, and died seven months later. Autopsy findings showed a hematogenous metastasis of the left precentral gyrus, measuring 3.0 x 2.5 x 2.5 cm.

[Clinical evaluation of long-term administration with terodiline hydrochloride for the treatment of patients with neurogenic bladder].

Terodiline HCl was administered in a long-term study to 20 patients with neurogenic bladder and pollakisuria. Its efficacy on urinary frequency and urinary incontinence was studied together with its safety and changes in blood concentration. The dosing period extended from 2 through 53 weeks (21 weeks on the average). The drug was found effective in 62% of diurnal urinary frequency patients, 71% of nocturnal urinary frequency, 73% of urinary incontinence, and 69% of nocturnal enuresis. Side effects of dizziness and nasal obstruction were seen in only one case. The drug was judged to be useful in 75% of the patients studied. Terodiline HCl showed no further increase in plasma concentration due to the long-term administration, and it disappeared from plasma within one to two months after the last dosing.