Breed distribution of SOD1 alleles previously associated with canine degenerative myelopathy.

BACKGROUND: Previous reports associated 2 mutant SOD1 alleles (SOD1:c.118A and SOD1:c.52T) with degenerative myelopathy in 6 canine breeds. The distribution of these alleles in other breeds has not been reported. OBJECTIVE: To describe the distribution of SOD1:c.118A and SOD1:c.52T in 222 breeds. ANIMALS: DNA from 33,747 dogs was genotyped at SOD1:c.118, SOD1:c.52, or both. Spinal cord sections from 249 of these dogs were examined. METHODS: Retrospective analysis of 35,359 previously determined genotypes at SOD1:c.118G>A or SOD1:c.52A>T and prospective survey to update the clinical status of a subset of dogs from which samples were obtained with a relatively low ascertainment bias. RESULTS: The SOD1:c.118A allele was found in cross-bred dogs and in 124 different canine breeds whereas the SOD1:c.52T allele was only found in Bernese Mountain Dogs. Most of the dogs with histopathologically confirmed degenerative myelopathy were SOD1:c.118A homozygotes, but 8 dogs with histopathologically confirmed degenerative myelopathy were SOD1:c.118A/G heterozygotes and had no other sequence variants in their SOD1 amino acid coding regions. The updated clinical conditions of dogs from which samples were obtained with a relatively low ascertainment bias suggest that SOD1:c.118A homozygotes are at a much higher risk of developing degenerative myelopathy than are SOD1:c.118A/G heterozygotes. CONCLUSIONS AND CLINICAL IMPORTANCE: We conclude that the SOD1:c.118A allele is widespread and common among privately owned dogs whereas the SOD1:c.52T allele is rare and appears to be limited to Bernese Mountain Dogs. We also conclude that breeding to avoid the production of SOD1:c.118A homozygotes is a rational strategy.

Xylan deposition on secondary wall of Fagus crenata fiber.

Delignified and/or xylanase-treated secondary walls of Fagus crenata fibers were examined by field emission scanning electron microscopy. Microfibrils with a smooth surface were visible in the innermost surface of the differentiating fiber secondary wall. There was no ultrastructural difference between control and delignified sections, indicating that lignin deposition had not started in the innermost surface of the cell wall. There was no ultrastructural difference between control and xylanase-treated sections. Microfibrils on the outer part of the differentiating secondary wall surface had globular substances in delignified sections. These globular substances disappeared following xylanase treatment, indicating that these globules are xylan. The globular substances were not visible near the inner part of the differentiating secondary wall but gradually increased toward the outer part of the secondary wall, indicating that xylan penetrated into the cell wall and continuously accumulated on the microfibrils. Mature-fiber secondary walls were also examined by field emission scanning electron microscopy. Microfibrils were not apparent in the secondary wall in control specimens. Microfibrils with many globular substances were observed in the delignified specimens. Following xylanase treatment, the microfibrils had a smooth surface without any globules, indicating that the globular substance is xylan. These results suggest that cellulose microfibrils synthesized on the plasma membrane are released into the innermost surface of the secondary wall and coated with a thin layer of xylan. Successive deposition of xylan onto the cell wall increases the microfibril diameter. The large amounts of xylan that accumulated on microfibrils appear globular but are covered with lignin after they are deposited.

Role of the putative membrane-bound endo-1,4-beta-glucanase KORRIGAN in cell elongation and cellulose synthesis in Arabidopsis thaliana.

A temperature-sensitive, elongation-deficient mutant of Arabidopsis thaliana was isolated. At the non-permissive temperature of 31 degrees C, the mutation impaired tissue elongation; otherwise, tissue development was normal. Hypocotyl cells that had established cell walls at 21 degrees C under light-dark cycles ceased elongation and swelled when the mutant was shifted to 31 degrees C and darkness, indicating that the affected gene is essential for cell elongation. Analysis of the cell walls of mutant plants grown at 31 degrees C revealed that the cellulose content was reduced to 40% and the pectin content was increased to 162% of the corresponding values for the wild type grown at the same temperature. The increased amounts of pectin in the mutant were bound tightly to cellulose microfibrils. No change in the content of hemicellulose was apparent in the 31 degrees C-adapted mutant. Field emission-scanning electron microscopy suggested that the structure of cellulose bundles was affected by the mutation; X-ray diffraction, however, revealed no change in the crystallite size of cellulose microfibrils. The regeneration of cellulose microfibrils from naked mutant protoplasts was substantially delayed at 31 degrees C. The recessive mutation was mapped to chromosome V, and map-based cloning identified it as a single G-->A transition (resulting in a Gly(429)-->Arg substitution) in KORRIGAN, which encodes a putative membrane-bound endo-1,4-beta-glucanase. These results demonstrate that the product of this gene is required for cellulose synthesis.

Identification of novel hypocholesterolemic peptides derived from bovine milk beta-lactoglobulin.

This study was designed to clarify the mechanisms of hypocholesterolemic action of beta-lactoglobuline tryptic hydrolysate (LTH) and to identify the novel hypocholesterolemic peptide derived from LTH by screening using Caco-2 cells and animal studies. Serum and liver cholesterol levels were significantly lower in rats fed LTH than in those fed casein tryptic hydrolysate (CTH). The present study suggests that the inhibition of micellar solubility of cholesterol which causes the suppression of cholesterol absorption by a direct interaction between cholesterol mixed micelles, and LTH in the jejunal epithelia is part of the mechanism underlying the hypocholesterolemic action of LTH. Though no one could trace the hypocholesterolemic peptide to any protein origin, we identified, for the first time, a novel hypocholesterolemic peptide, Ile-Ile-Ala-Glu-Lys (IIAEK). Surprisingly, the present study provides the first direct evidence that a new hypocholesterolemic peptide derived from beta-lactoglobuline can powerfully influence serum cholesterol levels and exhibit a greater hypocholesterolemic activity in comparison with that of medicine, beta-sitosterol, in animal studies.

Developmental toxicity of indium chloride by intravenous or oral administration in rats.

Pregnant rats were treated with a single intravenous or oral administration of indium chloride (InCl3) on day 9 of pregnancy and their fetuses were examined for growth and malformation on day 20 of pregnancy. By intravenous administration, fetal weight was significantly decreased and the incidences of fetal mortality and malformation were significantly increased at 0.4 mg In/kg. Fetal malformations of the tail and digits, e.g., kinked tail, brachyury, and oligodactyly, were observed at high incidences. By oral administration, similar tendencies in the fetal effects were observed, but there were no significant differences compared to the control even at 300 mg In/kg. Indium concentrations in the serum of pregnant rats showed low bioavailability of indium by oral administration. It was concluded from these results that indium showed teratogenicity in rats. Oral treatment with indium may be developmentally toxic at 300 mg In/kg, but this is difficult to state with certainty given the limited number of animals that were used in this study.

Serum cholesterol reduction and cholesterol absorption inhibition in CaCo-2 cells by a soyprotein peptic hydrolyzate.

The serum cholesterol level in rats was significantly decreased in a group fed on a soyprotein peptic hydrolyzate (SPH) when compared with a group fed on a casein tryptic hydrolyzate (CTH). The fecal excretion of total steroids was significantly greater with rats fed on the SPH diet when compared with the CTH diet. The results of CaCo-2 studies clearly suggest that the suppression of cholesterol absorption in the intestinal epithelia is part of the mechanism for the hypocholesterolemic action induced by SPH.

The influences of casein phosphopeptides on metabolism of ectopic bone induced by decalcified bone matrix implantation in rats.

The effect of casein phosphopeptides (CPP) on bone metabolism was studied in the ectopic bone induced by the implantation of decalcified bone matrix in rats. Forty-two Wistar male rats of 7 weeks old were fed low calcium diets (0.39% of calcium) with or without supplying 0.50% of CPP, or a control diet (0.91% of calcium) without CPP supplementation. After a 1-week preliminary period, each rat was subcutaneously implanted with 30 mg of demineralized bone matrix powder. Fourteen and 21 days after the implantation, the implants were harvested from 7 rats of each group. Calcium content in the graft was not significantly different among all groups on day 14. Subsequently, the content of calcium rapidly increased in the grafts irrespective of diets given. However, the graft of the CPP- group contained less calcium than the other groups and the calcium content was more in the control rats compared to the CPP+ animals on day 21. Alkaline phosphatase activity (an index of bone and cartilage calcification) was lower in the control group than in the CPP+ group on day 14. The enzyme activity subsequently decreased in the control group but the activity was not changed in the other groups. As a result, the activity of alkaline phosphatase was lower in the control animals than in the other rats on day 21. Tartrate-resistant cap phosphatase activity (an index of bone resorption) was higher in the CPP--group compared to the control on day 14. On day 21, the activity was higher in the CPP--group compared to the others.(ABSTRACT TRUNCATED AT 250 WORDS)

A comparison between ultrasonography and mammography, computed tomography and digital subtraction angiography for the detection of breast cancers.

Ultrasound (US) was compared with mammography (MMG), computed tomography (CT), and digital subtraction angiography (DSA) in its effectiveness to detect breast cancer masses and metastatic axillary nodes. Forty-seven breast cancer patients who all underwent MMG, US, CT, and DSA preoperatively in our institution between 1986 and 1990 were studied. US was able to detect tumors in all cases regardless of tumor size, whereas DSA detected T1-size tumors and MMG detected T2-size tumors in 40% and 64.7% of cases, respectively, being specifically inferior to US. It was found that MMG was least likely to detect papillotubular carcinoma, although microcalcification alone without a tumor mass on MMG improved detectability from 46.2% to 76.9%, according to the histological type. CT was found to be most sensitive to axillary node metastases (81.8%), followed by US (72.7%), but DSA was significantly unfavorable (42.9%). Thus, we concluded that US was superior to MMG, CT, and DSA for detecting breast cancer masses, but that CT was more advantageous than US, while DSA was of little value for evaluating axillary nodal status.

[Signification of liver metastases of colorectal cancer with special reference to recurrence in the residual liver after hepatic resection].

In the patients with liver metastases of colorectal cancer, pre-and post operative intra-arterial infusion chemotherapy was evaluated for prevention of recurrence in the residual liver after hepatic resection. Materials are sixty-five hepatectomized patients from May 1981 to 1992. Therapies were subdivided into five groups. I: pre-and postoperative non-therapy (n = 3); II: postoperative chemotherapy (n = 22); III: postoperative intra-arterial infusion chemotherapy (n = 12); IV: pre-operative intra-arterial infusion chemotherapy + postoperative chemotherapy (n = 15); and V: pre-and postoperative intra-arterial infusion chemotherapy (n = 13). In recurrence rate in the residual liver, I to IV groups showed as high as 50-100%. However, the disease-free survival rate was 100% in V group, revealing a significant difference between the other four groups. Accordingly, in order to prevent recurrence in the residual liver of hepatectomized patients with liver metastases of colorectal cancer and prolong the disease-free interval, we consider that pre-and post-operative intra-arterial infusion chemotherapy can be effective, compared to pre-or postoperative intra-arterial infusion chemotherapy alone.

[Complications and their management in intraarterial infusion chemotherapy].

Complications and its management were evaluated in intraarterial infusion chemotherapy for 188 patients with advanced carcinoma of the digestive organs from 1975 to Sept. 1991. Subjects were divided into four groups: Group I was 62 patients in whom the tip of the catheter without knots was established in the abdominal aorta via celiac axis, Group II consisted of 72 patients with the tip of the catheter without knots in the common hepatic artery. Group III had 35 patients with the tip of the catheter with knots (Anthron catheter) in the common hepatic artery. Group IV was 19 patients with the tip of the anthron catheter connected to the Infuse A-Port in the common hepatic artery. The most frequent complications seen among Group I, II and III were caused by catheter thrombosis (11.3%) in Group I, spontaneous dislodgement of catheter (26.4%) in Group II and extravasation (20%) in Group III. By using 16 gauge Toray Anthron catheter with Heparin coating on its inner and outer surfaces, the number of complications in Group I and II was kept smaller. Extravasation, on the other hand, has been less frequently seen in Group III by establishing the tip of the catheter at the branching site of the gastroduodenal artery from the common hepatic artery. Complications in Group IV (19 patients) were noted only in 3 patients, i.e., extravasation, subcutaneous necrosis and subcutaneous abscess, respectively. Therefore, we concluded that Group IV showed the most favorable intraarterial infusion chemotherapy with the most infrequent complications.

[Evaluation of modalities for recurrent breast cancer patients].

Of 342 breast cancer patients radically operated on in the Second Department of Surgery, School of Medicine, Chiba University during 1965-1988, treatment for 75 recurrent patients were evaluated by the initial modes of recurrence. The modes of recurrence were classified into distant metastases, local lymph node recurrence (axillary, parasternal and supraclavicular nodes) and chest wall recurrence according to the General Rules for Clinical and Pathological Recording of Breast Cancer. Of 75 recurrent patients, distant metastases were seen as common as 77.3%, followed by recurrences of local lymph nodes (14.7%) and chest wall (8.0%). The number of patients in each mode of recurrence increased in relation to increase in the size of tumor and the number of metastatic lymph nodes at the time of the first operation. Histologically, scirrhous carcinoma was most common in chest wall recurrence. 2-year disease-free survival rates of distant metastases, local lymph node recurrence and chest wall recurrence were 44.6%, 24.2% and 16.7%, respectively. 5-year survival of bone metastasis with chemo-endocrine therapy was as significantly favorable as 60%, compared to chemo- or radiotherapy alone (p less than 0.01). However, 5-year survival of lung metastasis with or without endocrine therapy revealed no significant difference. Local lymph node recurrence with the combination of resection, radio- and/or chemotherapy produced a trend toward showing more favorable survival than that without resection.

[Effects of combined therapy with irradiation, cisplatin and vindesine in lymph node recurrence of esophageal cancer].

The effects of combined therapy with irradiation, cisplatin and vindesine for lymph node recurrence of esophageal cancer was studied. The subjects were 95 patients with lymph node recurrence, who were divided into the following four treatment groups: Group I: Radiotherapy alone (R) (n = 31); Group II: cisplatin (CDDP) alone (n = 18); Group III: R + CDDP(n = 9); Group IV: R + CDDP + VDS (n = 10). The response rate (CR + PR) of Groups III and IV was 66.7% and 100%, respectively, which was significantly more favorable than 11.1% of Group II. The survival duration after recurrence was prolonged in the order of Group IV, Group III, Group II and Group I. In conclusion, combination therapy using R, CDDP and VDS will be effective for lymph node recurrence of esophageal cancer.

[Over two years survival of intra-arterial infusion chemotherapy in gastric cancer with liver metastases].

Between 1977 and April in 1989, long-term survivors (over two years) by intra-arterial infusion chemotherapy in gastric cancer patients with liver metastases were examined. The materials were 5 patients (4 synchronous, 1 metachronous metastases) among 21 P0H (+) gastric cancers. The extent of liver metastases shows 1 H1 and 4 H2. Reduction surgery was performed in 4 H2 patients (2 S2 + 3, 1 S4, 1 S6) and postoperative intra-arterial infusion chemotherapy via the catheter in the common hepatic artery was done to control the residual liver metastases. Continuous intra-arterial infusion chemotherapy with the regimen of FML (5-FU, MMC, Lentinan) revealed 100% response rate (3 CR, 1 PR). In a patient with metachronous metastases, PR was obtained with MA (MMC, ADM) + one-shot intra-arterial infusion of LAK cells. Among 5 patients, one with synchronous metastases has survived 35 months, followed by a patient who died after 32 months and two patients who died after 27 months. A patient with metachronous metastases has survived for 24 months.

[Combination of radiotherapy with cisplatin in esophageal cancer].

Combination of radiotherapy with Cisplatin was performed in order to improve the results obtained with radiotherapy alone in the treatment of esophageal cancer. The therapeutic effect of this regimen and administration of Cisplatin were analyzed in 7 primary and 8 recurrent esophageal cancer patients from April 1983 to February 1985 in N.I.R.S. Cisplatin 10 mg/m2, without hydration and diuresis, was given daily followed by radiotherapy of 2 Gy daily for five consecutive days. The course was performed for two consecutive weeks and repeated after a one-week withdrawal of Cisplatin alone. Total doses of Cisplatin were 200 mg and the total dosage of radiotherapy including fast neutrons was TDF 110 to 120. Cisplatin 30 to 50 mg/m2 was continued monthly for maintenance every four weeks after the second course. Response rates in primary and recurrent cases were 71.4% and 62.5% respectively. Administration of Cisplatin 10 mg/m2 daily was useful since nausea and vomiting rarely appeared and bone marrow toxicity and renal dysfunction as side effects were mild and reversible. In the combination of radiotherapy with Cisplatin, it seemed most effective to perform radiotherapy immediately after Cisplatin administration, considering the change in serum concentration.

Gallium induces reduction of negative charge of the cell membrane and redifferentiation of cancer cells.

In order to obtain an insight into the mechanism of experimentally induced redifferentiation of cancer cells, changes in negative charge of the cell membrane were examined by cytopherometry following in vitro exposure of the cancer cells to non-radioactive gallium. Reduction of the negative charge was observed at 24 hr and later. The gallium ions must have neutralized the negative charge of the cancer cell membrane to the normal range. This event may contribute in some way to cancer cell redifferentiation.

Semiquantitative 67Ga scanning of lung cancer by scintiscoring: its usefulness in diagnosis of localization, therapeutic effectiveness, radiation pneumo-mediastinitis and recurrence.

Attempts were made to attain a semiquantitative evaluation of therapeutic effectiveness of lung cancer by scintiscoring of the liver as well as the tumor lesion. A relative counting ratio of tumor to liver was computed, and this was found useful: in areas of active tumorous growth, higher ratios were obtained. The ratios returned to the control levels following irradiation with or without an anti-inflammatory regimen including prednine. The technic is simple, and yet invaluable especially when the tumor locates in the mediastinum escaping from radiological detection. Satisfactorily frequent repetition would be competent enough in early diagnosis of developing radiation pneumo-mediastinitis and regrowths in the mediastinum and fibrotic abnormal shadows as well. The principle would be best rewarded by use of a whole-body scanner equipped with a digital computer.