A radial flow bioreactor (RFB) is used for a three-dimensional perfusion culture of hepatocellular carcinoma (HCC) cells and renal cells, to create a bioartificial liver and kidney. The cylindrical reactor is filled with porous cellulose microcarrier. RFB can be characterized as a system in which the medium flows from the periphery toward the center of the reactor, thereby delivering an adequate supply of oxygen and nutrients to cells at the center as well as at the periphery. HCC cells incubated in the RFB system at high density maintained viability for long periods of time. Proximal tubular cells (LLC-PK1) as well as HCC cells, but not human immortalized mesangial cells (HMC) were cultured in the RFB for more than 14 days. The mRNA expression of some enzymes involved in the urea cycle, cytochrome P450s in HCC cells, and the 1-alpha-hydroxylase (CYP27B1) in LLC-PK1 cells was higher than that in monolayer cultures. These results suggest that the RFB system composed of HCC cells or renal cells may be useful for a bioartificial liver and kidney.

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Comparison between nonpulsatile and pulsatile machine perfusion preservation in liver transplantation from non-heart-beating donors.

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Alanine Transaminase↗

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Construction of chimaeric processed immunoglobulin genes containing mouse variable and human constant region sequences.

The specificity of monoclonal antibodies provides a powerful diagnostic and therapeutic tool in investigating human neoplasia. Radiological scanning and immunotherapy with mouse tumour-specific monoclonal antibodies have been applied to patients with some success, but a major problem is the neutralization of the mouse antibody induced by repeated administration of heterologous antibodies. To avoid or reduce such immune reactions, chimaeric immunoglobulins consisting of mouse variable (V) and human constant (C) regions can be synthesized. We have constructed a recombinant retrovirus DNA carrying genomic heavy-chain (H) variable-diversity joining (VH-D-JH) and C gamma 1 genes from different species and show here that the chimaeric intervening sequences are spliced out precisely. This procedure provides a useful method to construct the chimaeric mouse-human immunoglobulin gene to be expressed in Escherichia coli, yeast and animal cells. Unexpectedly, a hidden splice donor site in the 5'-flanking region of a human VH gene is used in place of the donor site of the leader sequence exon, resulting in the formation of the V region without the leader sequence.

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PubMedDate not supplied

Determination of carboxyhemoglobin in heated blood--sources of error and utility of derivative spectrophotometry.

The cause for discrepancies in results from different methods of the carboxyhemoglobin (HbCO) analysis on the blood from bodies of burn victims was investigated. Blood samples with 0, 50, and 100% carbon monoxide (CO) saturation were heated at various temperatures for some time and then analyzed. Carboxyhemoglobin content was determined by the fourth-derivative spectrophotometric method and compared with results from the usual two-wavelength method. For total hemoglobin measurement, the fourth-derivative technique and cyanmethemoglobin method were used. Turbidity in blood samples, which occurred when samples were heated above 50 degrees C, affected the analysis. At about 70 degrees C, coagulation and hemoglobin degeneration occurred accelerating the errors of determined values. The fourth-derivative technique, however, proved to be independent of the turbidity and would be useful for the analysis on the blood without hemoglobin degeneration.

Carboxyhemoglobin↗

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Myenteric and submucosal plexuses of the congenital aganglionosis rat (spotting lethal) as revealed by scanning electron microscopy.

A rat with a congenital aganglionosis develops a mega-small intestine and megacolon. This rat mutant has been used as an animal model for Hirschsprung's disease. The myenteric and submucosal plexuses of the rats were exposed by the trypsin-HCl digestion method and examined by scanning electron microscopy. In the control rats the myenteric and submucosal plexuses were present throughout the gut. However, in the mutant rats the myenteric plexus of duodenum was different from those of control rats; the mesh of plexuses varied both in size and shape. There were no myenteric or submucosal plexuses in the distal ileum or colon.

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