Human lymphocytes synthesize C-reactive protein

We obtained evidence for the synthesis and secretion of C-reactive protein (CRP) by peripheral mononuclear cells in culture. Human mononuclear cells isolated from peripheral blood, after depletion of platelets, were cultured in giutamine-depleted RPMI 1640 supplemented with [³H]glutamine in the presence of 10-O-tetradecanoyl-phorbol-13-acetate (TPA). Anti-CRP antiserum was added to the culture medium, and the resultant immunoprecipitate was analyzed in SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The immunoprecipitate consisted of CRP, heavy and light chains of IgG, and only the CRP protein band had radioactivity, indicating that CRP was synthesized by mononuclear cells. In the populations of mononuciear cells, T-cell preparations mainly synthesized CRP, under stimulation of a factor derived from activated monocytes. Studies using the inhibitors of phospholipid metabolism suggested that generation of the monocyte factor was relevant to metabolites of an arachidonate cascade.     

A new method of vascular anastomosis by CO2 laser: experimental and clinical study

It is difficult to maintain the long-term patency after conventional anastomosis especially for the small caliber vessels. Since 15 years we have performed aortocoronary bypass with suture materials for the patients with ischemic heart disease. There are some problems in maintaining the long-term patency of the bypass grafts. Low energy CO2 laser was utilized to make vascular anastomosis with a few stay sutures. Vascular anastomoses (side-to-side, end-to-end, end-to-side) were carefully made by CO2 laser in the regions of the femoral arteries and veins, the carotid arteries and jugular veins in dog. A-C bypass was also successfully carried out between the internal mammary artery and the left anterior descending artery under the beating heart in experiment. Outputs of 20-40 mW and irradiation times of 6-12 sec/mm were optimal conditions for anastomosis of the small caliber vessels. There were no problems in the intensity and the healing of the anastomotic sites in comparison with the conventional suture method. On the basis of these excellent experimental results a low energy CO2 laser was employed clinically for vascular anastomosis of the peripheral vessels in 28 patients with angina pectoris or chronic renal failure and cardiac failure. There were no complications such as bleeding and suture line aneurysm after surgery. In conclusion, vascular anastomosis by laser might be recommended in performing with safety and rapidity for small caliber vessels.     

Antigen-Driven Clonal Accumulation of Peritoneal γδ T Cells in vivo

How the clonality of γδ T cells changes in response to exogenous antigens is uncertain. Here we analyzed kinetics of Vγ1.1 and Vγ2 T cell clonality after intraperitoneal injection of purified protein derivatives (PPD) by the heterogeneity of the third complementarity determining region (CDR3) length in Vγ1.1-Jγ4-Cγ4 and Vγ2-Jγ1-Cγ1 junctions. The V-J junctions were analyzed in intrahepatic lymphocytes (IHL), spleen cells, and peritoneal exudate cells (PEC) by polyacrylamide gel electrophoresis. γδ T cells expressing Vγ1.1 and Vγ2 genes were heterogeneous in normal mice. Accumulation of specific Vγ1.1 T cell clones was transiently detected 7 days after the injection in PEC, but no accumulation was observed in IHL and spleen cells. The accumulated clones disappeared by 4 weeks. Transient accumulation of Vγ2 T cell clones was also observed in PEC at the early phase after the injection. These results suggest that γδ T cells with specific TCR respond to PPD and temporary accumulate in the peritoneal cavity, but not in liver and spleen.     

Stability of messenger RNA in postmortem human brains and construction of human brain cDNA libraries

We studied stabilities of poly(A)⁺-RNA in postmortem mouse and human brains for up to 12 hours. The yields of total RNA were not changed significantly during postmortem periods either in mouse brains or human brains. Cell-specific cDNA probes were used to evaluate postmortem stability of poly(A)⁺-RNA in each cell type in the central nervous system. We used neuronspecific enolase (NSE), S-100β (S-100), and myelin-associated glycoprotein (MAG) for molecular markers of neuron, astrocyte, and oligodendrocyte, respectively. There was no detectable degradation of mRNAs coding for NSE, S-100, and MAG during the postmortem periods on Northern blot hybridization analyses. These results indicate that intact mRNAs expressed in neuron, astrocyte, or oligodendrocyte can be isolated from postmortem brains for up to 12 hours after death. Using poly(A)⁺-RNA thus isolated from two postmortem human brains, we constructed directional cDNA libraries and demonstrated the presence of full-length cDNAs for NSE, S-100, and MAG on Southern blot hybridization analysis. The present data should encourage studies on altered gene expressions in human brain in various neurologic diseases.     

An alternative method of vascular anastomosis by laser: experimental and clinical study

In vascular surgery, it is now very difficult to maintain the long-term patency after a conventional vascular anastomosis, especially for small-caliber vessels. A low-energy CO2 laser was experimentally employed to make a vascular anastomosis with only a few sutures. Subsequently, it could be confirmed that optimal conditions for vascular anastomosis by laser were 20-40 mW in output and 6-12 sec/mm in irradiation time. On the other hand, pressure tolerance test as well as tensile strength test and microscopic examinations at the sites of anastomoses by laser were compared with the conventional suture method. There were no significant differences between laser and suture methods. On the basis of the excellent results of this study, the laser was clinically applied for anastomoses of the peripheral vessels in 35 patients. The first clinical laser application in the world was successful in a 44-year-old female patient with chronic renal failure in 1985. All patients are doing well without any complications from vascular anastomosis by laser. From these experimental and clinical studies, it can be concluded that anastomosis by laser should be recommended for small-caliber vessels such as aortocoronary bypass surgery.     

The neuropathogenesis of Borna disease virus infection

Borna disease virus(BDV) is a noncytolytic, neurotropic RNA virus that causes a disease of the central nervous system(CNS) in several vertebrate species, including horses, sheep, cats and ostriches. Epidemiological studies using peripheral blood or brain samples revealed that BDV can infect humans and that it may be related with certain neuropsychiatric disorders. The unique genetic and biological properties of BDV indicate that BDV develops a persistent infection in the CNS. Furthermore, a line of recent evidences suggests that BDV infection causes direct effects on brain functions in the absence of immunopathology-related brain damage. In this review, we discuss about recent data regarding neuropathogenesis of BDV infections in animals and humans.     

The V–J Recombination of T Cell Receptor-γ Genes Is Blocked in Interleukin-7 Receptor–deficient Mice

IL-7R-deficient mice have severely impaired expansion of early lymphocytes and lack γδ T cells. To elucidate the role of IL-7R on γδ T cell development, we analyzed the rearrangements of TCR-α, β, γ, and δ genes in the thymus of the IL-7R-deficient mice. Southern blot analysis with a Jγ1 probe revealed that more than 70% of Jγ1 and Jγ2 alleles are recombined to form distinct Vγ1.2–Jγ2 and Vγ2–Jγ1 fragments in control mice. On the contrary, no such recombination was detected in the mutant mice. The rearrangements in the TCR-α, β, and δ loci were comparably observed in control and mutant mice. PCR analysis indicated that the V–J recombination of all the Vγ genes is severely hampered in the mutant mice. The mRNA of RAG-1, RAG-2, Ku-80, and terminal deoxynucleotidyl transferase (TdT) genes was equally detected between control and mutant thymi, suggesting that the expression of common recombination machinery is not affected. These data demonstrated that the V–J recombination of the TCR γ genes is specifically blocked in the IL-7R-deficient mice and suggested the presence of highly specific regulation for TCR γ gene rearrangement.