Hybrid cell lines which secreted antibodies to liver-specific membrane lipoprotein (LSP) were obtained by immunizing SMA and BALB/c mice with human LSP and fusing their splenocytes with the myeloma cell line P3-NSI/1-Ag4-1. The secretion of antibody to LSP (anti-LSP) was monitored by binding to a human hepatocellular carcinoma cell line, SK-Hep-1, which possesses surface membrane LSP, and to 125I-antimouse F(ab')2 antibody in radiobinding assay, and by reacting with 125I-LSP in double-antibody radioimmunoassay. From four separate cell fusions, seven secreting hybrids were cloned by dilutional techniques. Of these, four cell lines produced antibodies reacting with a wide variety of cells. The culture supernatants of the remaining three (6D6, 6G3 and 8F10) demonstrated the strongest binding activities against SK-Hep-1 among the various kinds of cell lines tested. However, binding with other cell lines, including renal cancer cells (SK-RC-6) and myeloid cell (HL-60) also occurred. Absorption test of ascitic fluids derived from 6D6 showed that ascitic fluids lost their capacity to bind to target SK-Hep-1 cells when absorbed with SK-Hep-1. Similarly absorption by SK-RC-6 and HL-60 removed almost all of the binding activity of ascitic fluids. Moreover, the binding activities of the ascitic fluids to SK-RC-6 and HL-60 were eliminated when absorbed with SK-RC-6, HL-60 and SK-Hep-1. The present study indicates that our LSP preparation contains nonspecific organ antigens, and although LSP exists on liver cell membrane, it is also found on the cell membrane of other organs albeit in less quantity.

Animals↗

PubMedDate not supplied

Isolation of a novel analgesic pentapeptide, neo-kyotorphin, from bovine brain.

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Amino Acid Sequence↗

PubMedDate not supplied

An antiserum to atrial natriuretic factor (ANF) cross-reacts with neurophysins in the hypothalamo-neurohypophysial system of rat brain.

The distribution of atrial natriuretic factor (ANF)-like reactivity was examined in rat brain and heart by immunohistochemistry. Immunostaining in heart was confined to atrial myocytes. In the hypothalamus, ANF-absorbable immunoreactivity was observed in magnocellular perikarya of the paraventricular and supraoptic nuclei, and in their projections to the neural lobe of the pituitary gland. No staining was seen in the preoptic or arcuate hypothalamic nuclei or in brain stem nuclei as previously reported by other investigators. The patterns of reactivity for ANF reported here is similar to that observed for neurophysins (NPs). Comparison of sequence data between rat ANF-28 and bovine NPs revealed three regions of 3 amino acid homology between these hypothalamic peptides. Preabsorption of the ANF antiserum with Affigel-coupled bovine NP I also resulted in complete elimination of all "ANF-immunoreactivity" in both atrium and hypothalamus. Cross-reactivity of the ANF antiserum with bovine NP I and II was further confirmed by Western blot analysis. Our findings suggest that ANF antisera can cross-react with NPs if they are directed against the shared antigenic epitopes; complete elimination of staining by preabsorption of the antibody with the immunogen, therefore, does not guarantee authenticity of localization. These observations may have relevance to an earlier study which reported on the existence of ANF-immunoreactivity in oxytocin neurons of the hypothalamus.

Amino Acid Sequence↗

PubMedDate not supplied

Postnatal ontogeny of catecholamine and somatostatin neuron systems in the median eminence of the rat as revealed by a colocalization technique.

The ontogenetic development of catecholamine (CA) and somatostatin containing nerve terminals in the rostral, central and caudal median eminence (ME) of the rat was investigated by combining fluorescence histochemistry and immunohistochemistry in the same tissue section. Somatostatin terminals were detected earlier in development than CA terminals and had already appeared in the lateral part of the external layer of the central ME by the 1st postnatal day. CA nerve terminals were first observed in the same region of the ME on the 7th postnatal day. At about this stage both types of terminals seemed to show early signs of a correlation in their distribution which became progressively closer as maturation proceeded. Their distribution reached a stable condition in density and pattern on the 21st postnatal day when the majority were found in the lateral part of the external layer of the central ME, in the basal part of the brain just dorsal to the tuberoinfundibular sulcus and in the upper and lower labia of the tuberoinfundibular stalk. They subsequently matured to the adult pattern of stabilized distribution and anatomical relationship. It was also established that on the 28th postnatal day the somatostatin immunoreactive terminals reached a greater concentration than at any other stage of development including that of the adult.

Aging↗

PubMedDate not supplied

High resolution DSA: experimental and clinical evaluation.

The DSA system has been upgraded for capabilities of slow scan video technique and progressive T.V. read-out of 1024 X 1024 matrix with 10 bits of depth. A square wave test pattern made of lead bar demonstrated moderate improvement in spatial resolution, but imaging of a Burger-Rose phantom revealed no significant increase in contrast resolution. Clinical study of various angiograms with intraarterial injections showed that there was slight improvement in the visibility of the vessels, especially small arterial branches, while there was no increase in visibility of the veins. There were no cases in which diagnoses were altered by application of high resolution DSA. With future improvements of the image intensifiers, DSA with 1024 X 1024 matrix may reveal its potential advantage, especially when coupled to larger image intensifiers such as 12 or 14 in.

Angiography↗

PubMedDate not supplied

Biplane digital subtraction angiography.

Biplane digital subtraction angiography was performed by placing a rapid switch between the anteroposterior and lateral T.V. cameras and the A-D converter. Alternate exposures from two X-ray tubes were employed. Excellent anteroposterior and lateral DSA was obtained especially for the brain and neck. Although there was more time required for positioning, biplane imaging saved the second or the third injection in many occasions. The technique has been especially valuable for intravenous DSA of the brain and the neck.

Angiography↗

PubMedDate not supplied

Hybrid digital subtraction angiography: initial clinical experience.

Hybrid digital subtraction angiography was performed after modification of a DSA system. Alternate high voltage (110-120 kV) and low voltage (65-70 kV) were obtained within short time intervals of 60-70 ms with a high voltage switching generator. Experimental phantom studies revealed the exposure dose per single image of hybrid subtraction was approximately 30% of that of conventional DSA. The SNR of hybrid subtraction was improved by increasing the exposure dose and application of postprocessing programs. Clinical application of intraarterial and intravenous DSA revealed that soft tissue artifacts were removed surprisingly well, but image quality decreased because of lowered SNR. Frame integration and matched filtering improved image quality of hybrid subtraction.

Angiography↗

PubMedDate not supplied

In vivo coordination structural changes of a potent insulin-mimetic agent, bis(picolinato)oxovanadium(IV), studied by electron spin-echo envelope modulation spectroscopy.

Bis(picolinato)oxovanadium(IV) [VO(pic)2] is one of the most potent insulin-mimetic vanadium complexes. To probe coordination structural changes of this complex in vivo and provide insights into the origin of its high potency, an electron spin-echo envelope modulation (ESEEM) study was performed on organs (kidney, liver and bone) of VO(pic)2- and VOSO4-treated rats. Kidney and liver samples from both types of rats exhibited a 14N ESEEM signal that could be attributed to equatorially coordinating amine nitrogen. The relative intensity of the amine signal was larger for the organs of the rat treated with the less potent VOSO4, suggesting that this amine coordination inhibits the insulin-mimetic activity. The spectra of kidney and liver from the VO(pic)2-treated rat contained a weak signal due to the picolinate imine nitrogen. This suggests that some picolinato species (including both the bispicolinato and a partially decomposed monopicolinato species) still exist in the organs as a minor species, where the proportions of the picolinato species to the total amount of the EPR-detectable VIVO species are estimated as 8-16% in the kidney and 12-24% in the liver. The picolinate ligand presumably serves to prevent VO2+ from being converted into the inactive amine-coordinated species. Bone samples from both types of rats exhibited an ESEEM signal due to 31P nuclei. The VO2+ in bone is therefore most likely incorporated into the hydroxyapatite Ca10(PO4)6(OH)2 matrix, which is consistent with the hypothesis that the bone-accumulated VO2+ is gradually released and transported to other organs as is Ca2+. No 14N signals were observed, even in the bone samples of the VO(pic)2-treated rats, indicating that vanadium uptake by bone requires complete decomposition of the complex.

Animals↗

PubMedDate not supplied

Synthesis and properties of an oligonucleotide modified with an acridine derivative at the artificial abasic site.

The synthesis of an oligodeoxynucleotide (ODN) modified with 2-methoxy-6-chloro-9-aminoacridine (Acr) at an abasic site is described. A stereochemically defined aminodiol, L-threoninol, was used to serve as artificial abasic nucleoside. The molecule was modified so as to be suitable for the standard phosphoramidite method and was incorporated into the interior of an ODN. In addition, N-hydroxysuccinimidyl N-[9-(6-chloro-2-methoxy)acridinyl]-6-aminocaproate has been synthesized for postsynthetic modification of the amino substrate of the L-threoninol moiety in the ODN. By using absorption spectroscopy, it is shown that oligo(dA) conjugated with acridine binds with complementary strand in a 1:1 ratio. The melting temperature showed that the nonmodified (abasic) duplex is destabilized as a result of lacking in base at the abasic site, but the covalently linked acridine ring compensates for the destabilization effect. The fluorescence quantum yield of the acridine ring was enhanced by connection to oligo(dA) and, further, by formation of a double-strand with the complementary ODN. The quantum yield is larger than that of intermolecular intercalation. The excitation spectra of Acr-ODN in the duplex is quite similar to the absorption spectra. The results indicate that the covalently linked acridine ring is selectively intercalated into the adjacent abasic site.

Acridines↗

PubMedDate not supplied

Effects of soy protein diet on the expression of adipose genes and plasma adiponectin.

Many studies have reported the cholesterol-lowering, anti-lipogenic, anti-obesity and anti-hypertensive effects of soy protein. Adipose tissue-specific plasma protein, adiponectin, has anti-atherogenic and anti-insulin-resistance properties. Here, we investigated the effects of soy protein diet on body fat composition, plasma glucose, lipid and adiponectin levels and expression of genes involved in glucose and fatty acid metabolism in obese KK-A y mice. Body weights and adipose tissue weights of mesenteric, epididymal, and brown fat were lower in mice on calorie-restricted diet containing soy protein isolate. Plasma cholesterol, triglyceride, free fatty acid, and glucose levels were also decreased by this diet. Body fat content and plasma glucose levels in mice on a soy protein isolate diet were still lower than those treated with an isocaloric casein-protein-diet. Among the genes related to glucose and fatty acid metabolism, adiponectin mRNA levels in adipose tissue and adiponectin plasma concentrations were elevated in mice on a calorie-restricted diet, although there were no significant differences between soy protein and casein protein groups. Our results indicate that that soy protein diet decreased body fat content and plasma glucose levels more effectively than isocaloric casein-protein diet in obese mice.

Adiponectin↗

PubMedDate not supplied

Extracorporeal hybridization of proximal renal tubular cells and an artificial membrane for the purpose of beta 2 microglobulin removal.

The present study was designed to develop a bioartificial kidney hybridized with LLC-PK1 cells for the purpose of beta 2microglobulin (B2M) removal. Cells were prepared in polystyrene culture wells; the bottom of each was covered with collagen fiber and the cells grown as monolayer cultures. Each well was inoculated with 1 ml cell suspensions with 0.0, 1.0 X 10(6), 2.0 X 10(6), or 4.0 X 10(6) cells/well. B2M was added to the culture system at a concentration of 80 micrograms/dl. Results were as follows: 1) Concentration of B2M in the supernatant of 4 X 10(6) cultivated LLC-PK1 cells was decreased from 79.32 +/- 2.41 micrograms/dl to 67.02 +/- 3.00 micrograms/dl (n = 6, p less than 0.01) at 2 hr; 2) Concentration of B2M in the supernatant of 1 X 10(6), 2 X 10(6), and 4 X 10(6) cultivated LLC-PK1 cells was 75.06 +/- 2.31 micrograms/dl (n = 6, p less than 0.05); 67.27 +/- 11.41 micrograms/dl (n = 6, p less than 0.001); and 67.02 +/- 3.00 micrograms/dl (n = 6, p less than 0.001), respectively, vs. 79.32 +/- 2.41 micrograms/dl in controls at 2 hr; 3) There was a significant positive correlation between the efficacy of B2M removal and cell number; and 4) These results suggested that it is possible to develop an artificial kidney hybridized with proximal tubular cells that possesses a high affinity for B2M removal.

Animals↗

PubMedDate not supplied

Development of continuous recirculating peritoneal dialysis using a double lumen catheter.

Continuous recirculating peritoneal dialysis (CRPD) was newly introduced to improve solute removal efficiency in conventional dialysis therapies such as hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD). In CRPD, a part of the dialysate in the peritoneal cavity was drained through a double-lumen catheter and purified by an extracorporeal dialyzer. Urea removal characteristics in CRPD were examined in a canine study. In this study, a recirculation-dialysis experiment using a dog weighing 9.0 kg was carried out under 100 and 200 ml/min of flow for recirculating and delivered dialysates, respectively. An FB-50H (Nipro Medical Industries, Ltd., Osaka, Japan) composed of cellulose diacetate membrane with 0.5 m2 of surface area and Dianeal-1.5 (Baxter Limited Laboratories, Tokyo, Japan) containing urea were used as the extracorporeal dialyzer and dialysate. Urea peritoneal and dialyzer dialysances (DBP and DBD) were 3.05 and 33.3 ml/min by computer simulation using a compartment model for CRPD. This DBP value can be estimated as 20.3 ml/min for a 60 kg human. From this result, time-averaged value for BUN over an 8 hr/day CRPD, combined with three exchanges/day as CAPD is estimated to be 34.3 mg/dl, which is much lower than 45.2 mg/dl for a 12 hr/week HD, or 53.0 mg/dl for conventional CAPD.

Animals↗

PubMedDate not supplied

Fast overlapping scans in cerebral computed angiotomography.

Three to four overlapping cerebral CT scans were obtained after the injection of a single bolus of contrast medium. The rapid sequential dynamic scanning was carried out with table increments, while there was peak iodine density in the carotid artery. The demonstration of cerebral vessel lesions (aneurysms, moyamoya disease, arteriovenous malformations, arterial narrowing, etc.) is easier and safer with this method as compared with the conventional multiple injection technique.

Adult↗