Plasma metabolite concentrations and hepatic enzyme activities in pregnant Romney ewes with restricted feeding.

Plasma metabolite concentrations and activities of enzymes related to energy metabolism in plasma, peripheral leukocytes and liver of pregnant Romney ewes with restricted feeding were measured to assess those metabolites and enzymes as indicators for evaluating metabolic conditions in the ewes. The body weights and plasma lactate concentrations of the low-feeding ewes (about 1.0 times maintenance) were significantly lower than those of the high-feeding ewes (about 1.5 times maintenance). There were no significant differences in plasma protein, glucose, triglyceride, cholesterol, pyruvate and immunoreactive insulin concentrations and plasma and leukocyte enzyme activities between both groups. Hepatic malate dehydrogenase activities were significantly higher and hexokinase activities were significantly lower in the low-feeding ewes than in the high-feeding ewes. Restricted feeding could maintain pregnancy and some plasma metabolites and peripheral leukocyte enzymes may be useful indicators for evaluating metabolic changes in ewes.

Changes in peripheral leukocytes enzymes activity and plasma metabolite concentrations in growing Holstein calves.

Plasma metabolite and immunoreactive insulin concentrations and activities of enzymes related to energy metabolism in peripheral leukocytes were measured in growing Holstein calves. A ratio of girth of abdomen divided by girth of thorax (A/T ratio) of calves was significantly elevated after weaning, and the A/T ratio maybe a good indicator to evaluate rumen development. Plasma glucose and free fatty acid concentrations were changed in calves accompanying change in feeding. Activities of lactate dehydrogenase with pyruvate as substrate (LDH-P) and hexokinase (HK) in cytosolic fractions of peripheral leukocytes decreased significantly after weaning the calves reflecting the change of energy source from milk replacer with high percentages of fat and glucose and lactose as absorbable carbohydrate to pelleted feed containing starch as less absorbable carbohydrate and roughage. Some peripheral leukocyte enzymes such as LDH and HK may be good indicators to evaluate changes in energy metabolism of growing calves.

Cloning and expression of a Porphyromonas gingivalis gene for protoporphyrinogen oxidase by complementation of a hemG mutant of Escherichia coli.

Porphyromonas gingivalis, a bacterium implicated in periodontal pathogenesis, has a growth requirement for iron protoporphyrin IX. By complementation with a P. gingivalis 381 chromosomal DNA library, we were able to isolate a clone that enhanced the poor growth of a hemG mutant of Escherichia coli. The DNA sequence analysis of this clone revealed three open reading frames (ORFs). ORF3 encoded a protein of 466 amino acids with a calculated molecular weight of 51 695 Da. The deduced amino acid sequence of the ORF3 gene had significant similarity to sequences of protoporphyrinogen oxidase (PPO) from Myxococcus xanthus (30% identical residues). When the ORF3 gene was overexpressed in E. coli, the extract had much higher PPO activity than a control extract, and this activity was inhibited by acifluorfen, a specific inhibitor of PPO. Thus, ORF3 was named PgHemG. Furthermore, several porphyrin-related genes, including hemD, hemN and hemH, were identified in the data bases on the websites available on-line. We postulated that a porphyrin biosynthetic pathway to heme from preuroporphyrin may be conserved in P. gingivalis.

Relationship between tooth loss and electrocardiographic abnormalities in octogenarians.

Recent reports have suggested that periodontal disease is a risk factor for coronary heart disease (CHD). However, little is known about the relationship between periodontal disease and CHD in the very elderly. Therefore, we evaluated the association between tooth loss and abnormal ECG findings in octogenarians. Of the 1,282 octogenarians in Fukuoka Prefecture, Japan, 697 participated. Oral examinations--including a tooth count and an assessment of the Community Periodontal Index--and a 12-lead ECG were performed. Logistic regression analysis revealed that individuals with < 20 teeth had increased prevalence of ST segment depression, and T-wave abnormalities, after we adjusted for gender, smoking, serum cholesterol and glucose, blood pressure, and body mass index. Compared with individuals with > or = 20 teeth, edentulous individuals had an increased prevalence of T-wave abnormalities and arrhythmias after adjustment for coronary risk factors. We conclude that tooth loss may be an independent predictor of abnormal ECG findings in octogenarians.

Double blind study of the effect of caffeine on the mental calculation.

The double blind study of the effect of caffeine on the mental calculation has been carried out in the practical exercise course for the undergraduate students of clinical pharmacology, in the Medical School, Kyushu University. The results obtained from 1997 to 1999 were summarized and evaluated.

Histological evaluation of allograft bone after acetabular revision arthroplasty: report of two cases.

We recently had the opportunity to take histological sections from two patients who underwent acetabular reconstruction in which allograft and ME Müller acetabular roof reinforcement rings were used. In one patient (case 1), histological sections of the chipped allograft were taken on two separate occasions from the same area, at 7 months, and at 3 years and 11 months after the bone graft. The histology of the chipped allograft showed necrosis at 7 months, but almost normal morphology of trabecular bone formation at 3 years and 11 months after the bone graft. In the other patient (case 2) histological sections of the block allograft and chipped allograft were taken at 1 year and 8 months after the bone graft. The block allograft showed only a small amount of admixture of newly formed bone with the necrotic bone, while the chipped allograft showed a large amount of newly formed bone, with only a small amount of necrotic bone remaining. Therefore, we principally use chipped allograft for acetabular reconstruction, in order to achieve early and complete graft incorporation. If a block allograft is used in a weight-bearing area, it should be protected from excessive load by using an acetabular reinforcement device.

Distribution and localization of cells and collagens in the proliferated intima of arterially implanted autovein grafts.

We examined the microscopic features and distribution of collagens in the hyperplastic intima of arterially implanted autovein bypass grafts under conditions of a reduced blood flow with a poor distal outflow. Vascular anastomosis was made using 7-0 nonabsorbable polypropylene sutures (PP group), or absorbable polydioxanone sutures (PDS group). On the contralateral limb, an autovein bypass graft was performed under normal flow conditions (NF group). The thickness of the intima in the NF group was approximately 50 microm throughout the duration of the study, while in the PP and PDS groups, intimal hyperplasia progressed to 290+/-112 microm and 267+/-123 microm, respectively, at 13 months after grafting. Collagen accumulated significantly in both the PP and PDS groups; types IV and V collagen in particular increased considerably in the deep layer. Regardless of the suture materials, the progression of intimal hyperplasia was considered to be closely related to the poor distal outflow to be and caused by the proliferation of myofibroblasts and active production of collagen. The increase in types IV and V collagen, particularly in the deep layer of the hyperplastic intima, was due to development of numerous vasa vasora in this region.

Angioarchitecture and blood circulation in focal nodular hyperplasia of the liver.

BACKGROUND/AIMS: We sought to clarify important unresolved points regarding angioarchitecture and blood circulation in focal nodular hyperplasia. METHODS: Twenty-nine surgically resected focal nodular hyperplasia lesions were examined histologically, immunohistochemically, and radiologically. In three autopsy cases, red- and blue-colored gelatin was injected into the hepatic artery and the portal vein, respectively, to demonstrate the vasculature in focal nodular hyperplasia. RESULTS: Histologically, no orientation with respect to portal tracts and central veins was evident in any lesion. Within lesions, vessels were classified as: (i) anomalous arteries in the fibrous septa, (ii) capillaries in the fibrous septa, or (iii) venous vessels located mainly in the parenchyma. Vessels and sinusoids adjacent to fibrous septa were stained for CD 34 and von Willebrand factor. The anomalous arteries were connected to the capillaries. Capillaries in the fibrous septa were connected to sinusoids adjacent to fibrous septa. Venous vessels were connected to central or hepatic veins surrounding the lesions. Intranodular sinusoids were connected to the sinusoids in the surrounding normal liver. Red-colored gelatin, injected at autopsy into the hepatic artery, appeared not only in the anomalous arteries but also in capillaries and in sinusoids adjacent to the fibrous septa of the lesion. Angiography clearly depicted hepatic veins located near the lesions in nine cases. Computed tomography during arterial portography disclosed no portal blood flow in the lesions. CONCLUSIONS: In focal nodular hyperplasia, arterial blood flows from the anomalous arteries via the capillaries into sinusoids adjacent to fibrous septa. The blood in the sinusoids drained to the hepatic vein either directly or via perinodular sinusoids.

Wear of bipolar hip prostheses.

A method was developed to take radiographs showing the inner articulation of bipolar hip prostheses. By this method, wear was measured in 68 hips whose inner head diameter was 22 mm. Average annual wear rate was 0.17 mm. Osteolysis was observed in 25 hips (37%) and there was no difference between the annual wear rate of hips with and without osteolysis. Studying 19 retrieved prostheses, abrasion of the rim was deeper in hips with osteolysis than those without it. Wear rate of the inner articulation in bipolar hip prosthesis is much larger than that in Charnley's prosthesis, as linear penetration into the articulation surface reduces the motion range of the inner articulation and this increases impingement and advances rim abrasion.

Epidermoid cyst of the spleen with CA19-9 or carcinoembryonic antigen productions: report of three cases.

True splenic cyst is a relatively rare disease, and the majority of the cases are classified as epidermoid cysts. Three cases of epidermoid cysts in the spleen or accessory spleen were studied using an immunohistochemical technique and staining for mucin. In case 1, serum carcinoembryonic antigen (CEA) and CA19-9, and in cases 2 and 3, serum CA19-9, before surgery were markedly elevated, and these levels decreased postoperatively. This strongly indicates the relationship between the increase of tumor marker levels and the presence of the epidermoid cyst. In addition, stratified squamous epithelium in the resected tissues of cases 1 and 2 was positive for anti-CEA antibody and anti-CA19-9 antibody, and that of case 3 was positive for anti-CA19-9 antibody. This strongly supports CEA or CA19-9 production in the squamous epithelium.

Development of vasa vasorum in the arterially implanted autovein bypass graft and its anastomosis in the dog.

Using stereomicroscopy, light microscopy, and scanning electron microscopy, we investigated the development of vasa vasorum in the proliferated neointima of the autovein graft and its anastomoses implanted in the canine femoral artery against a background of poor distal runoff. In the stereomicroscopic examination, a microfil silicone rubber compound (MF) was injected transluminally or via perivascular vasa, and the vascular specimen was prepared for clearing by immersion in a methyl-salicylate solution. Vessel interstices filled with MF were found adjacent to the suture materials within 5 days of grafting. Fourteen days after implantation, luminally originating vasa vasorum were often visible in the neointima along the suture line and distributed into the media and adventitia connecting to the original vasa vasorum. At 6 months or more after grafting, many orifices of luminally originating vasa vasorum were seen along the suture line of both proximal end-to-end and distal end-to-side anastomoses and distributed into the thickened neointima forming a vasa network when the neointima had proliferated to over 250 microm in depth. On the other hand, some clefts filled with MF were found in mural thrombi deposited on the vascular sinus of the graft within 5 days, and these appeared to be one of the sources of luminally originating vasa vasorum on the graft distant from the suture line. Moreover, the development of numerous vasa vasorum was constantly demonstrated in the neointima when it had proliferated to over 250 microm in depth.

[A case of cat scratch disease identified by an elevated Bartonella henselae antibody level using enzyme immunoassay].

A 68-year-old male was admitted to our hospital because of fever and a 2-week history of inguinal adenomegaly. Since he owned a cat, cat scratch disease was suspected. But it was necessary to distinguish cat scratch disease from lymphoma type adult T-cell leukemia because he showed a high level of antibody against HTLV-1. An excisional biopsy of the inguinal node was performed. Histopathologic examination revealed abscess-forming granulomatous lymphadenitis compatible with cat scratch disease. A Warthin-Starry silver stain showed pleomorphic bacilli in the lymph node. So we confirmed a serological response to Bartonella henselae, the causative agent of cat scratch disease, using enzyme immunoassay (EIA). The IgG antibody level to B. henselae was positive at 42 EIA Unit before treatment. After treatment with intravenous cefepime and oral tosufloxacin, his physical symptoms improved and the antibody level decreased to less than 12 EIA Unit. EIA was very useful for diagnosis of this case. Serology to B. henselae may replace traditional diagnostic criteria for cat scratch disease.

[The significance of the medical research study: hemodynamic and pathophysiologic characteristics in peripheral arterial reconstruction].

In cases of peripheral arterial reconstructive surgery, it is important to prevent postoperative early and late occlusion of the reconstructed artery. The postoperative early outcome of the reconstructed artery were related to electromagnetically determined flow waveform of the reconstructed artery. Flow waveform was classified in 5 types, i. e. types O, I, II, III, and IV. The prognosis of the reconstructed arteries with types O and I was excellent. Type II showed no postoperative early occlusion but late occlusion did occur in 31%. Type III and IV were all failed within 48 hours postoperatively. Correlation between prognosis of the reconstructed artery and flow waveform was established by the intraluminal velocity profile of blood flow, using a specially designed flow wave simulation pump system. Postoperative late occlusion in arterial reconstruction often occurs in cases with poor distal runoff and caused by intimal hyperplasia of implanted autovein graft and its distal end-to-side anastomosis. The features of intimal hyperplasia of the implanted autovein graft differed from those at the distal end-to-side anastomosis; the former being related to active proliferation of smooth muscle cells while the latter displayed an excessive proliferation of fibroblasts and collagen fibers.

Femoral component wear in retrieved hip prostheses.

We retrieved 159 femoral heads at revision surgery to determine changes in surface configuration. Macroscopic wear of the head was observed in three bipolar hip prostheses as a result of three-body wear. There was a considerable change in surface roughness in the internal articulation of bipolar hip prostheses. Roughness in alumina heads was almost the same as that in new cobalt-chromium heads. The annual linear wear rate of polyethylene cups with alumina heads was less than that of cups with cobalt-chromium alloy heads. Polyethylene wear was increased in the prostheses which had increased roughness of the head.

Hepatocellular carcinoma with sarcomatous change.

We studied clinicopathological and immunohistochemical features of hepatocellular carcinoma (HCC) with sarcomatous change among surgically resected HCC cases. Among 308 HCC cases, 6 (1.9%) showed partial (5 cases) or entire (1 case) sarcomatous change. Only 1 case received preoperative anti-cancer therapy among the 6 cases, suggesting that sarcomatous change is not always related to anti-cancer therapy. Immunohistochemically, the sarcomatous components were positive for albumin and alpha 1-antichymotrypsin and negative for alpha-fetoprotein in all cases. Expressions of intermediate filament-related proteins (e.g., cytokeratin and vimentin) in the sarcomatous components were heterogeneous. The sarcomatous components showed stronger expressions of p53 protein and/or mouse double minute-2 protein, and higher proliferating cell nuclear antigen labeling index, suggesting their high proliferative activity and high malignant potential.

[A case report of surgical treatment of constrictive pericarditis with coronary artery disease].

We performed the concomitant operation for constrictive pericarditis and coronary artery disease in an octogenarian. A 82-year-old male was hospitalized with dyspnea, edema of the lower extremities and pleural effusion on chest X-ray film. Cardiac catheterization revealed constrictive pericarditis and 75% stenosis of left anterior descending artery. Extensive pericardiectomy was performed including posterior wall of left ventricle and left atrium under the beating heart by using femoro-femoral partial bypass. Single CABG with a saphenous vein graft was performed following pericardiectomy. Postoperative cardiac catheterization showed good recovery of hemodynamics and patency of the bypass graft. Postoperative course was uneventful. The patient was discharged on twenty fifth postoperative day.

A new human hepatocellular carcinoma cell line (HAK-2) forms various structures in collagen gel matrices.

We established a new human hepatocellular carcinoma (HCC) cell line, designated HAK-2, from a surgically resected HCC of a 57-yr-old Japanese man. The patient's tumor consisted of 5 different histological features in a single nodule: well-differentiated HCC with trabecular pattern; and moderately differentiated HCC with 4 different patterns (i.e., trabecular, pseudoglandular, solid, and scirrhous). Morphologically, HAK-2 cells on a plastic dish showed oval-shaped nuclei and large flat, polygonal eosinophilic cytoplasm and proliferated in a monolayered sheet with a population doubling time of 36.8 hours. Meanwhile, various structures, such as compact, trabecular, and tubular arrangements, were induced in HAK-2 cells cultured in type I collagen gel matrix. Also, HAK-2 cells in vitro underwent spontaneous apoptosis more frequently than other HCC cell lines examined. HAK-2 cells secreted various plasma proteins including albumin into the culture medium. Chromosome and flow cytometric analyses revealed that HAK-2 had many structural abnormalities with human karyotype and a single aneuploid cell population with a DNA index of 3.7, respectively. These findings suggest that HAK-2 is a new human HCC cell line representing two morphological characteristics; (1) formation of various structures in the presence of extracellular matrix and (2) frequent spontaneous apoptosis in vitro.