Autologous blood transfusion for kidney transplant recipients.

Autologous blood transfusion (ABT) is rarely employed in patients with end-stage renal disease (ESRD); these patients are usually anemic. Since 1998, we have attempted ABT for ESRD patients undergoing living-related kidney transplantation. Among 20 patients enrolled in this study the preoperative hemoglobin and hematocrit levels were 10.0 +/- 1.2 mg/dL (range, 8.1-11.7) and 30.0 +/- 3.7% (range, 24.7-34.3), respectively. Blood volume collected on each occasion was 235.7 +/- 57.7 mL (range, 200-400), and the number of blood collections was 2.45 +/- 0.9 (range, 1-4). Total collected volume was 567.5 +/- 157.5 mL (range, 400-800). Symptomatic hypotension was seen in two patients, but vital signs recovered spontaneously. No other problems related to blood collection were observed. Allogeneic transfusion was need in only one patient (5%). ABT was safe and efficacious in ESRD patients scheduled for living-related kidney transplantation.

Shedding of porcine circovirus into colostrum of sows.

The purpose of the present study was to investigate porcine circovirus (PCV) shedding into the milk of sows. Colostrum was collected from 33 sows. PCV1 was isolated from four of 33 milk whey samples. PCV1 DNA was detected by polymerase chain reaction in three of these samples and in three of 10 milk cell samples. PCV2 was also isolated and detected from every single milk whey sample. These results showed that PCV1 and PCV2 were shed into the milk of sows and suggest that PCV can be transmitted to offspring by an oral route through milk.

[Auditory cortical response to monaural stimulation as detected by functional magnetic resonance imaging].

In order to confirm the crossed-innervation between auditory cortex and the ear that receives monosyllabic sound, the auditory cortical response to monaural monosyllabic stimulation as detected by functional magnetic resonance imaging (fMRI) was investigated in six normal hearing subjects. Stimulus amplitude averaged 95 dBSPL at the distal end of the audio system. A series of 440 echo planar images was acquired during the acoustic stimulation within the four OFF-ON cycle paradigm. Five image series with 10 slices were collected within each OFF or On period. Each scanning session began with four baseline images before the OFF-ON paradigm. Monosyllabic sounds were presented monaurally during the ON period at a rate of one monosyllable/sec. Functional MRI data were analyzed with SPM99b software (Statistical Parametric Mapping). The background scanner noise averaged 97dBSPL. The selicon ear plug and headphone as acoustic shields attenuated the noise as much as 17 dB. A broad and intense auditory cortical response was observed bilaterally in response to monaural monosyllable stimulation. Sound presentation to the right ear was followed by a larger response in the left auditory cortex than in the right, and left ear stimulation evoked a larger response in the right auditory cortex than in the left. This pattern was consistent in all subjects examined. The primary auditory cortex responded to monosyllabic words presented to the contralateral ear. The results confirmed the crossed-innervation between the auditory cortex and ear for listening to monosyllables. Functional MRI is a useful tool for investigating auditory cortex function, if the scanner noise is adequated controlled.

Spontaneous galactosylation of agalactoglycoproteins in colostrum.

We have found that spontaneous galactosylation of GlcNAc residues occurs in bovine colostrum, but not in dialyzed colostrum, without adding UDP-Gal as a donor substrate. UDP-Gal was shown to be present in bovine colostrum at a level ranging from 200 to 600 microM. When a tracer UDP-[(14)C]Gal was added to the dialyzed colostrum together with a Gal beta1,4-specific beta-galactosidase, remarkable incorporation of radioactivity into 24-28 kDa and 33 kDa RCA1-positive glycoproteins was demonstrated by SDS-PAGE/autoradiography. Some 100-140 kDa agalactoglycoproteins of a CHO mutant cell line were also galactosylated on a blotted membrane by the incubation in the colostrum.

[Long-term results of Feldmann's osteoplastic approach for chronic middle ear disease].

Between 1978 and 1981, Feldmann's osteopathic approach was often used to manage chronic middle ear disease. In this procedure, the superior and posterior segment of the ear canal wall was cut after complete mastoidectomy, removed temporarily and re-positioned in the previous position after handling the diseased focus in the tympanic isthmus area. Forty-one cases (24 cases of non-cholesteatomatous chronic otitis media and 17 cases of cholesteatoma) were followed and long-term results of this procedure were studied with regard to re-operative findings following this procedure. Among the 41 patients, 13 (31.7%) required revision surgery because of cholesteatoma formation, infection etc. Seven of these 13 patients (53.8%) required revision surgery because of cholesteatoma formation after this procedure. None of these 7 cases appeared to involve residual cholesteatoma. The most important problem is that 3 of the 7 patients showing cholesteatoma formation had non-cholesteatomatous chronic otitis media before this procedure. In other words, the Feldmann's osteoplastic approach may iatrogenically induce cholesteatoma formation in non-cholesteatomatous chronic otitis media. The re-operative findings indicated that the re-positioned canal wall in this procedure may have small bony defects or bony erosion, inducing pocket formation through these defects to create a new cholesteatoma. Although recent literature concerning tympanoplasty recommends posterior canal wall reconstruction using cartilage, bone, ceramic material or bone-pate rather than the canal wall down method, careful follow-up should be continued with regard to pocket formation and/or cholesteatoma formation.

Detection of reverse transcriptase activity by enzyme-linked immunosorbent assay in human immunodeficiency virus type 1.

An enzyme-linked immunosorbent assay (ELISA) using biotin-labelled oligo-dT primer and digoxigenin (Dig)-dUTP was designed to measure the reverse transcriptase (RT) activity of human immunodeficiency virus type 1 (HIV-1). The ELISA system involves the selective detection step of a newly synthesized cDNA by two specific bindings, biotin-streptavidin binding and alkaline phosphatase (AP)-conjugated anti-Dig-Dig binding, and the enzymatic amplification step to increase coloring generated by AP. This method was used to measure the activity of RT in the culture supernatants of peripheral leukocytes obtained from four anti-HIV-1-positive persons cocultivated with those from four anti-HIV-1-negative persons. RT activity was detected in all of four anti-HIV-1-positive culture supernatants but not in those cultivated with anti-HIV-1-negative supernatants alone. Thus, our improved ELISA for detection of HIV-1 appears to be sensitive enough and useful for routine laboratory work. This non-radioactive method will also be useful for detecting other retroviruses and for screening of RT inhibitors.

Peptide:N-glycosidase activity found in the early embryos of Oryzias latipes (Medaka fish). The first demonstration of the occurrence of peptide:N-glycosidase in animal cells and its implication for the presence of a de-N-glycosylation system in living organisms.

The recent discovery of free oligosaccharides typical for the complex type of glycan chains terminating with a free di-N-acetylchitobiosyl structure in certain fish eggs and early embryos (Ishii, K., Iwasaki, M., Inoue, S., Kenny, P. T. M., Komura, H., and Inoue, Y. (1989) J. Biol. Chem. 264, 1623-1630; Seko, A., Kitajima, K., Iwasaki, M., Inoue, S., and Inoue, Y. (1989) J. Biol. Chem. 264, 15922-15929; Inoue, S., Iwasaki, M., Ishii, K., Kitajima, K., and Inoue, Y. (1989) J. Biol. Chem. 264, 18520-18526) led us to find an enzyme responsible for detachment of N-linked glycan chains from glycoproteins by hydrolyzing the beta-aspartyl-glucosylamine linkage in Oryzias latipes embryos. The enzyme, peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine amidase or peptide:N-glycosidase (PNGase), was partially (2090-fold) purified, and the reaction site at which this enzyme acts was specified by analysis and identification of the reaction products. This is the first demonstration showing PNGase in animal sources, although the presence of PNGases was reported in a variety of plant extracts and bacteria. Thus, the commonality of this type of enzyme is now demonstrated, and the possible physiological role of PNGase in de-N-glycosylation as a basic biologic process is proposed.

Stimulation of human peripheral blood polymorphonuclear cell iodination by lignin-related substances.

Lignin is a heterogenous natural product composed of phenylpropane units and is usually associated with hemicellulose in its native state. Until now little attention has been paid to the potential therapeutic utility of lignified products. Natural lignified products are demonstrated in the present study to stimulate iodination significantly (incorporation of radioactive iodine into an acid-insoluble fraction) of human peripheral blood polymorphonuclear cells (PMN). This stimulation was significantly inhibited in the presence of myeloperoxidase inhibitors. These materials were almost completely deprived of their stimulation capacity by treatment with NaCIO2, but this capacity was not affected by severe treatment with H2SO4 or trifluoroacetic acid. Similar stimulating activity by chemically defined tannin-related polyphenolic compounds was observed. Degradation products or component units of lignin, and natural antitumor polysaccharides and their chemically modified derivatives (introduced with negatively or positively charged groups) and polysialoglycoproteins had little or no activity. The results indicate the importance of a polymerized phenolic structure for the stimulation of PMN iodination. Possible physiological relevance of the stimulation of iodination by lignified substances is discussed.

Mitogenic activity of pine cone extracts against cultured splenocytes from normal and tumor-bearing animals.

An acidic pine cone extract, Fr. V. of Pinus parviflora Sieb. et Zucc. significantly stimulated DNA synthesis of isolated splenocytes from both mice and rats, but only marginally affected the DNA synthesis of leukemic cell lines. The maximum stimulation level attained by Fr. V slightly exceeded that of plant lectins, whereas much weaker stimulating activity was found in natural and chemically modified antitumor polysaccharides, sialic acid-rich glycoproteins, and polyphenolic compounds such as lignin and tannic acid. In mice with subcutaneously transplanted sarcoma-180, responses of splenocytes against Con A declines in the terminal stage of tumor development, whereas responses against Fr. V remained relatively constant throughout all periods of tumor progression. The suppression of Fr. V activity by acetylation or methylation suggests the importance of the hydroxyl group in the expression of its stimulation activity.

Inhibition of herpes simplex virus infection by tannins and related compounds.

Several chemically defined plant extracts were investigated for their antiviral action on herpes simplex virus (HSV-1, HSV-2)-infected African green monkey kidney cells and human adenocarcinoma cells, using a plaque formation assay. Among them, the monomeric hydrolyzable tannins, oligomeric ellagitannins and condensed tannins, having galloyl groups or hexahydroxydiphenoyl groups, had the most potent anti-HSV activity. Their 50% effective doses (0.03-0.1 microgram/ml) were by two-three orders of magnitude lower than their 50% cytotoxic doses (greater than 10 micrograms/ml). On the other hand, gallic acid, neutral polysaccharides, chemically modified (N,N-dimethylaminoethyl-, carboxymethyl-, and sulfated-) glucans, sialic acid-rich glycoproteins, and uronic acid-rich pine cone polysaccharide showed little or no activity. Using radiolabeled virus particles, we demonstrated that the anti-HSV effect of the tannins is due to inhibition of virus adsorption to the cells.

Effect of pentazocine and related compounds on the lipid composition of Ehrlich ascites tumor cells.

The effect of pentazocine and its related compounds on the lipid composition of Ehrlich ascites tumor cells was examined. Ehrlich tumor cells were suspended in Hanks balanced salt solution (pH 7.4) supplemented with 2% bovine albumin (2 x 10(6) cells/ml) and incubated at 37 degrees C for 120 min with and without 10(-3) M of pentazocine, morphine or dihydrocodeine. After incubation, the tumor cell lipids were extracted with chloroform-methanol (2:1, v/v), and they were analyzed quantitatively by the dichromate reduction procedure of Amenta. The tumor cells treated with pentazocine contained lower levels of triglycerides and cholesterol esters as compared with the tumor cells incubated alone. The amounts of triglycerides and cholesterol esters in the tumor cells treated with and without pentazocine were about 82 and 23 mg/10(10) cells and 182 and 55 mg/10(10) cells, respectively (P less than 0.01), whereas there was only a slight difference in the contents of these neutral lipids between the tumor cells treated with and without (10(-3) M morphine or dihydrocodeine. In addition, the fatty acid pattern of triglycerides and cholesterol esters from the pentazocine-treated tumor cells differed markedly from that of the corresponding lipids from the tumor cells incubated alone.

Acetyl-coenzyme-A carboxylase in cultured hepatocytes. Effects of exogenous fatty acids on the content, synthesis and degradation of the enzyme.

Studies were made on the content, synthesis and degradation of acetyl-coenzyme-A carboxylase in JTC-25 - P3 cells, hepatocytes which can be maintained in a protein-free and lipid-free chemically defined medium. The addition of corn oil or fatty acid to the medium resulted in a decrease in the activity level of the enzyme without impairing the viability of cells. All the fatty acids tested exhibited this effect, although linoleic acid and oleic acid were more effective than palmitic acid, stearic acid and arachidonic acid. Immunochemical titration and Ouchterlony double-diffusion analysis indicated that the decrease in the activity level of the enzyme observed in cells incubated in medium supplemented with fatty acid can be ascribed to a reduction of the quantity of the enzyme. Isotopic leucine incorporation studies with the use of immunochemical techniques demonstrated that this reduction of the enzyme content is due to a decrease in the rate of synthesis of the enzyme. The rate of degradation of the enzyme was essentially unaffected, the half-life being 25 and 28 h, respectively, in cells incubated in the presence and absence of fatty acid. It was shown that most of the isotopic fatty acid added to the medium was incorporated into cellular phospholipids, while a very small portion of it was recovered in triglyceride and nonesterified fatty acid.