[Laboratory diagnosis of protozoan and parasitic diseases. 2. Helminthiasis. c. Echinococcosis].
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Echinococcosis↗
Biomedical subjects
K Furuya
Publications and source records attributed to K Furuya.
Essential light chain exchange in smooth muscle myosin.
The exchange of 17-kDa essential light chain (LC17) in smooth muscle myosin was carried out by incubating myosin with a 10-fold molar excess of exogenously added LC17 over the corresponding endogenous light chain in the presence of trifluoperazine and 4.5 m ammonium chloride. Porcine aorta myosin contains two kinds of LC17 isoform, LC17nm and LC17gi, while chicken gizzard myosin contains only one kind of LC17 isoform. As LC17gi can be separated from LC17nm and gizzard LC17 by urea-gel electrophoresis, LC17nm in aorta myosin and LC17 in gizzard myosin were exchanged with LC17gi and LC17gi in aorta myosin was exchanged with LC17nm, and the degree of exchange was estimated by urea-gel electrophoresis. Under the optimal conditions (6 and 10 degrees C for aorta and gizzard myosin, respectively), nearly 90% of exchange, which is close to the theoretical value, was achieved for the former combinations, and a slightly lower exchange was obtained for the latter. The LC17-exchanged myosins contained stoichiometric amounts of the heavy and light chains and retained the original nature in the phosphorylation-dependent actin-activated ATPase activity, 6S-10S conformational transition, and filament assembly.
Actin Cytoskeleton↗
Non-GABAergic effects of midazolam, diazepam and flumazenil on voltage-dependent ion currents in NG108-15 cells.
Non-gamma-Aminobutyric acid (GABA)-mediated effects of benzodiazepines (BZs) have not been widely investigated. However, there is significant evidence in the literature to suggest that several experimental and clinical observations are inconsistent with the commonly accepted GABAergic mechanisms of action for these drugs. The purpose of the present study was to explore electrophysiological effects of midazolam, diazepam and a specific BZ antagonist, flumazenil, using patch-clamp techniques in NG108-15 cells which do not express the GABAA receptor. Midazolam and diazepam decreased Na+, K+ and Ca2+ currents in a dose-related manner. Ca2+ currents were reduced more significantly by diazepam than by midazolam. Flumazenil showed no effects on voltage-dependent ion currents. GABA by itself showed neither effects on the membrane potential nor these ion currents. Midazolam and diazepam, but not flumazenil, exhibited effects on voltage-dependent ion currents in cultured neurons.
Animals↗
Enhanced strength in cemented stem fixation using adhesive acrylic cement as a metal coating material.
One cause of aseptic loosening of cemented total hip arthroplasty is mechanical weakness at the interface between the metal stem and the cured bone cement. Adhesive acrylic bone cement containing 4-methacryloyloxyethyl trimellitate anhydride (4-META) was applied as a metal coating material to increase the strength of the cemented fixation. The 4-META cement has 2-3 times greater tensile bond strength to metals than does commercial acrylic bone cement. The shear strength of the coated metals fixed with bone cement was approximately 4 times greater in SUS-304 and 3 times greater in titanium (Ti) alloy than those of uncoated metals, and this strength did not decrease after 1 week's immersion in saline. The coating process using the 4-META cement can be performed at normal room temperature, so that metal stems for bone cement fixation could be coated during the course of an operation resulting in potentially improved results of total hip arthroplasty.
Biocompatible Materials↗
Temperature dependency of calcium responses in mammary tumour cells.
Changes of intracellular calcium concentration ([Ca2+]i) induced by the extracellular application of ATP and bradykinin in mouse mammary tumour cells (MMT060562) were investigated by image analysis of fluo-3 fluorescence at 24 degrees C and 35 degrees C. ATP (0.1-100 microM) and bradykinin (0.1 nM-1 microM) induced the increase of [Ca2+]i at both temperatures and Ca(2+)-depletion did not affect these [Ca2+]i responses. Both [Ca2+]i responses became more sensitive at 35 degrees C than at 24 degrees C. A clear latency of [Ca2+]i increase after the application of the agonists was observed, and it changed with the concentration of the agonist. As concentrations of ATP or bradykinin became lower, the latency and rise time became longer. At higher concentrations, the latency and rise time approached a constant value. The latency shortened remarkably at 35 degrees C. These results suggested the involvement of a regenerative or threshold process in the [Ca2+]i responses in mammary tumour cells.
Adenosine Triphosphate↗
Establishment of a primary culture of Echinococcus multilocularis germinal cells.
This study was designed to establish an in vitro primary culture of germinal cells of Echinococcus multilocularis, a parasite that causes alveolar echinococcosis of the liver (AEL). We also investigated the temperature-dependency of the cultured cells. The germinal cells, which originated from a human lesion, were cultured by an original fluid-suspension method at 25 degrees C or 37 degrees C for 4 weeks. Anchorage-dependent and -independent cells were observed by light microscopy, transmission electron microscopy, and immunocytochemistry to confirm their origin. Cell number and viability were examined by immunocytochemistry and mitochondrial exclusion test. The cultured cells were also inoculated into jirds (Meriones unguiculatus) to evaluate metacestode formation. Morphology and immunocytochemistry showed that the cultured cells were typically germinal cells. The cell number declined gradually over the 4-week culture period, but viability remained at 50% at 3 weeks. These findings were not associated with either of the two culture temperatures; moreover, host-associated cells were not noted in the cultured cells at 25 degrees C. The implanted cells formed metacestodes in the jird peritoneal cavity, and their histology demonstrated mature and typical alveolar-type echinococcal cysts. We successfully established an in vitro primary culture of germinal cells. This should contribute to future studies, and, hence, a better outcome for patients with AEL.
Animals↗
Contribution of mass screening system to resectability of hepatic lesions involving Echinococcus multilocularis.
The prognosis for patients with alveolar echinococcosis of the liver (AEL) is excellent when the lesion is completely resected. Early detection of the disease and subsequent resection of the lesion are thus indispensable; however, the usefulness of screening systems is now controversial. This study was designed to compare screened and non-screened patients according to stage classification and to re-evaluate the effect of screening. We studied a total of 82 patients (63 screened and 19 non-screened). The stage classification showed a significant intergroup difference (P < 0.002). The largest tumors ranged from 30 to 100 mm, and there was a significant intergroup difference (P < 0.0014). Ultrasonography showed even small lesions in the screened patients. The complete resection rate was 74.6% for the screened patients, and 21.1% for the non-screened patients, showing a significant difference (P < 0.0001). The rate of unresectable lesions was higher in the non-screened patients (32%) than in the screened patients (11%), showing a significant difference (P < 0.04). The present screening system contributes to early detection and subsequent resection of AEL, leading to a better outcome.
Animals↗
Intraperitoneal dissemination probably caused by needle biopsy of alveolar echinococcosis of the liver: experimental study.
Alveolar echinococcosis of the liver (AEL) is a parasitosis with a potential for malignant tumor-like behavior. The disease is diagnosed by a combination of serologic tests, diagnostic images, and the histology of needle biopsy specimens. It remains unresolved whether the biopsy induces subsequent troubles. We designed this study to investigate critical problems after needle biopsy of AEL lesions using an experimental model. Five samples were prepared from the resected lesions of AEL patients: (A) 10% suspension of trypsin digests of the minced lesion; (B) 10% suspension of mesh-filtered sediment of the minced lesion; (C) 10% sediment suspension after washing the nonminced lesion; (D) supernatant after centrifuging intracystic fluid; (E) 10% sediment suspension after centrifuging intracystic fluid. A 1-ml aliquot of each sample was injected intraperitoneally into jirds (gerbils) or cotton rats, respectively. The animals were sacrificed 12 weeks later, and intraperitoneal metacestodes were observed. All samples except D developed metacestodes, and their histologies were all lesions of typical alveolar echinococcosis. These results suggest that a needle biopsy may cause intraperitoneal dissemination or tracial implantation of the parasites along the track of the needle.
Animals↗
Involvement of P2-purinergic receptors in intracellular Ca2+ responses and the contraction of mammary myoepithelial cells.
Mammary myoepithelial cells were isolated and cultured to characterize their properties. The intracellular calcium concentration (Cai2+) was measured using the ratio of fura-2 fluorescence at 340 nm to that at 360 nm (F340/F360), and the contraction was simultaneously monitored by the change of fluorescence intensity at 360 nm (F360). Cultured myoepithelial cells retained their contractile machinery as in the intact tissue. NBD-phallacidin fluorescence, which marks F-actin, and electron microscopy showed abundant bundles of microfilaments in the cytoplasm. Oxytocin (> 0.1 nM) induced an increase in Cai2+ and contraction. The amplitude and time course of the Cai2+ increase were not markedly affected in the Ca2+-free solution. Nifedipine (10 microM) did not affect the response to oxytocin. ATP (>1 microM) induced an increase in Cai2+ and contraction. The response to ATP was not affected by Ca2+ removal, but was suppressed by suramin (100 microM), an antagonist of P2-purinergic receptors. The order of potency of nucleotides to increase Cai2+ was ATP = ADP > UTP > UDP. These findings indicate the presence of P2-purinergic receptors in mammary myoepithelial cells. The results suggest that stimulant-induced Cai2+ increases and contractions are due to release of Ca2+ from intracellular stores in these cells. In addition to the hormonal action of oxytocin, extracellular nucleotides may function as paracrine agents to contract myoepithelial cells in the mammary gland.
Actins↗
A monoclonal antibody to astrocytes, subepithelial fibroblasts of small intestinal villi and interstitial cells of the myenteric plexus layer.
A monoclonal antibody was developed, using cultured subepithelial fibroblasts of rat duodenal villi as the antigenic material, by in vitro immunization. Hybridomas were selected on cryosections of rat brain and small intestine using indirect immunofluorescence techniques. The monoclonal antibody, termed 8E1, was very useful to label GFAP-positive astrocytes in the adult rat brain, subepithelial fibroblasts of intestinal villi and Thuneberg's fibroblast-like interstitial cells associated with the myenteric plexus layer. These cells are known to be stellate in shape, forming a cellular network with extended cell processes that communicate via gap junctions. Immature astrocytes such as radial fibers were not immunolabeled with mAb-8E1. Immunoelectron microscopy revealed clustered gold labeling of 8E1 antigen on the intermediate filaments of cultured astrocytes and cultured subepithelial fibroblasts. This staining pattern was different from the continuous and linear gold labeling of GFAP and vimentin. MAb-8E1 did not immunoblot purified human brain GFAP nor bovine lens vimentin, and so was not neutralized by preabsorption with these molecules. Immunoblot analysis after SDS-PAGE showed that the antigen recognized by mAb-8E1 was a Mr 50K protein in the rat cerebra, and a Mr 56K protein in the cultured subepithelial fibroblasts. This study showed that astrocytes and some types of stellate cells share distinct antigenic properties in the components of their intermediate filaments.
Animals↗
Efficacy of amino acid infusion for improving protein metabolism after surgery: a prospective randomized study in patients undergoing subtotal gastrectomy.
BACKGROUND: Appropriate regimens of peripheral parenteral nutrition (PPN) have been proposed for the improvement of protein metabolism after surgery. When evaluating the efficacy of administered nutrients, it is vital to consider the severity of surgical stresses to avoid confounding effects of the trauma on the postoperative metabolism. This study was designed to evaluate protein-sparing regimens through PPN in patients who had undergone subtotal gastrectomy. STUDY DESIGN: Patients hospitalized at our institutes for gastric cancer were randomly divided into the following five groups and received PPN for 7 days after surgery: 1. G group (n = 9), 200 g glucose (per day); 2. AG group (n = 10), 54 g amino acids + 150 g glucose; 3. AGG group (n = 9), AG + 110 g glucose; 4. AGF group (n = 10), AG + 40 g fat; and 5. AGL group (n = 7), 58 g amino acids + 60 g glycerol. Biochemical studies were done before and after surgery. RESULTS: In comparison to G group patients, AG group patients showed less negative cumulative nitrogen balances. No significant differences in cumulative nitrogen balances were observed between AGG, AGF, and AGL groups. Restoration of the reduced serum rapid turnover protein occurred earlier in the AGL group than in either the AGG or the AGF groups. Hyperglycemia, glucosuria, and hyperinsulinemia were prominent in the AGG group, and less prominent in the AGL group. Marked ketosis together with an increase in serum-free fatty acid levels was found in the AGL group. CONCLUSIONS: These results suggest that in patients who have undergone major elective surgery, infusion of amino acid solutions is advantageous for improving protein metabolism after surgery, and nonprotein energy source and intake are not essential when combined with amino acid solutions for improving nitrogen balance after surgery.
Adult↗
Growth-associated protein GAP-43 detection in the neuronal somata following middle cerebral artery occlusion in the rat.
We first detected growth-associated protein (GAP-43) immunoreactivity in the neuronal somata following middle cerebral artery occlusion in the rat. Four days after the middle cerebral artery occlusion, GAP-43 immunoreactivity was detected in some cortical neurons of the ischemic penumbra at the level of the hippocampus.
Animals↗
Genetic factors in lung disease: atopy and bronchial asthma.
Atopy defined as high IgE responsiveness has now been subject to genetic studies at the molecular level owing to the development of a great number of DNA markers over the human genome. Either by linkage analysis or by association study strong candidate genes of atopy have been proposed to be located on chromosome 11q13 and 5q31 where high-affinity IgE Fc receptor beta subunit and allergy-associated cytokines, respectively, have been mapped. Meanwhile, we found a novel association between one of alleles of D11S97, an anonymous DNA marker on 11q13, and high total serum IgE in a large number of Japanese general population and atopic family members. However, failure to replicate linkage or association studies by different investigators suggest polygenic nature of atopy. In addition to the genes regulating IgE synthesis, the requirement of local (pulmonary) genetic factors in the development of bronchial asthma have been speculated. Linkage analysis suggested possible existence of gene(s) regulating susceptibility and/or clinical characteristics of bronchial asthma also on chromosome 5q. One of the candidate is beta 2-adrenergic receptor gene polymorphism. Mutated gene transfection studies suggested functional significance of some polymorphisms and clinical evaluations have revealed their contribution to airway responsiveness and severity of asthma.
Asthma↗
Genetic factors in lung disease. Part II: Lung cancer and angiotensin converting enzyme gene.
The recent progress in molecular biology has led to the elucidation of pathogenesis of lung cancer. The development of a lung cancer requires multiple genetic changes, consisting of the activation of oncogenes, including the K-ras and myc genes, and of inactivation of tumour suppressor genes, including the Rb, p53 and CDKN2 genes. Knowing the specific genes undergoing such changes should be useful as biomarkers for the early detection of cells destined to become malignant. Moreover, such genetic changes could be targets of newly designed drugs and gene-based therapy. Although the angiotensin I-converting enzyme was originally discovered in equine plasma, it has been recognized in various organs and cells other than vascular endothelial cells. This enzyme is also known to have wide substrate specificity to many peptides. The definite roles of angiotensin converting enzyme (ACE) in the respiratory system are largely unknown. Recent progress in molecular biology of the ACE, however, gives us a good chance to look over the significance of ACE in respiratory diseases as well as cardiovascular disorders. In this review, we show the recent advances in the basic studies of the ACE and refer to its clinical application.
Gene Expression Regulation, Neoplastic↗
Parathyroid-hormone-related protein in human milk and its relation to milk calcium.
Parathyroid-hormone-related protein (PTHrP) is found in high concentrations in human milk. One of the possible physiological roles of PTHrP may be the regulation of calcium transport through the mammary gland. The relationship between milk PTHrP and Ca concentrations was evaluated in lactating women in the present study. Milk PTHrP has been quantified by immunoradiometric assay (IRMA) and radioimmunoassay (RIA). The IRMA detects PTHrP 1-72, and the RIA carboxyterminal region of PTHrP (C-PTHrP). Milk PTHrP 1-72 and C-PTHrP concentrations did not significantly change by breast-feeding. Milk PTHrP 1-72 changed significantly by the time of day, in contrast to C-PTHrP. Thus, PTHrP 1-72 rather than C-PTHrP may reflect an acute change in mammary PTHrP secretion. In lactating women in whom milk PTHrP and calcium concentrations were serially measured at 5-day intervals up to 26-30 days after delivery, milk PTHrP 1-72 progressively increased with duration of lactation. In contrast to PTHrP 1-72, milk C-PTHrP decreased during this period. And milk PTHrP 1-72 levels inversely correlated with C-PTHrP levels. Milk C-PTHrP but not PTHrP 1-72 levels correlated significantly with milk Ca concentrations. Although it is not certain whether the correlation between milk C-PTHrP and Ca levels is causally related, it may be that fragments of PTHrP detected as C-PTHrP are involved in the regulation of Ca transport through the mammary gland in women.
Calcium↗
[Trial for transdermal administration of sulfonylureas].
Sulfonylureas are widely used as oral hypoglycemic drugs in the treatment of non insulin dependent diabetes mellitus (NIDDM). Since sulfonylureas are usually taken for a long period, the compliance of the patients is very important. Therefore, for the improvement of the compliance of the patients, the development of a transdermal dosage form of sulfonylureas was attempted in this study. Glibenclamide (GLI) or chlorpropamide (CHL) was chosen as a principal agent and ointments were prepared by mixing 5% of GLI or CHL with a FAPG ointment base. Penetration and shearing stresses of the ointments were determined as physical characteristics of the ointments. There was no obvious difference of characteristics between the GLI ointment, or the CHL ointment and the FAPG ointment base. In drug release tests, the CHL ointment showed better release of the drug than the GLI ointment. In both ointments, comparatively rapid release of drug was observed in the initial 1 h, and continuous slow release was observed thereafter. When the ointments were applied on the back of male Wistar rats and the plasma glucose level was measured, both CHL and GLI ointments gave lower blood glucose levels than the control (FAPG base). At the all measuring points, the GLI ointment brought about significantly lower blood glucose levels than the control (p < 0.01). Thus, it was demonstrated that sulfonylureas were absorbed through the skin and lowered the blood glucose levels. The results suggest the possibility of transdermal administration of sulfonylureas for the treatment of NIDDM.
Animals↗
[Social support of chronically-ill children and healthy children].
The purpose of this study was to describe characteristics of social support of chronically-ill children and to compare social support of chronically-ill children with those of healthy children. Subjects consisted of 204 chronically-ill children aged between 9 and 18 years old and 544 healthy children aged between 10 and 17 years old. Results of this study were as follows: 1. In both chronically-ill children and healthy children, parents' support score and teachers' support score were significantly higher in elementary school children than high school students. In both groups, friends' support score was significantly higher in girls than boys. 2. Chronically-ill children reported significantly higher scores of parents' support, friends' support and teachers' support than those of healthy children. 3. Parents' support score was significantly lower in children with diabetes than those with asthma and cancer. 4. The total score of social support and teachers' support score were significantly lower in children with employed mothers than those with unemployed mothers. 5. The total social support score of children who participated in non-sports extracurricular activities was significantly higher than those of children who did not participate in non-sports activities.
Adolescent↗
Role of calcium ions in the potentiation by isoproterenol of carbachol-induced ionic currents and secretion activity in rat salivary glands.
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Animals↗