Intrahepatic Delivery of Glutathione by Conjugation to Dextran

Glutathione was covalently attached to dextran (T-40) by the CNBr activation method. The compound obtained was a water-soluble powder containing 10 (w/w%) glutathione, which was gradually released from the conjugate in aqueous media. Mice depleted of glutathione by treatment with buthionine sulfoximine, a potent inhibitor of -glutamylcysteine synthetase, exhibited a significant increase in hepatic glutathione level after intravenous injection of the conjugate. In mice given a lethal dose of acetaminophen, the survival rate increased progressively with coadministration of the conjugate, whereas little improvement was found when free glutathione was given. The conjugate maintained the serum transaminase activities at lower level after acetaminophen administration. These findings suggest that the dextran conjugate of glutathione is transported into hepatic cells and is intracellulary hydrolyzed to free form, which protects mice from hepatotoxicity due to acetaminophen.     

Characterization of factor XII Tenri, a rare CRM-negative factor XII deficiency

Factor XII Tenri (Y34C), a rare cross-reacting material (CRM)-negative factor XII deficiency, was identified in a 71-yr-old Japanese woman with angina pectoris. In the patient's plasma, factor XII activity and antigen levels were only 1.6% and 5.0%, respectively, of those seen in a normal subject. Immunoblot analysis showed that the secreted factor XII Tenri existed not only as a monomer (76 kDa), but also in complexes with apparent molecular weights of approximately 115, 140, 190, 215, and 225 kDa. After reduction with 2-mercaptoethanol, the factor XII Tenri contained in the complexes was completely converted to monomeric form on immunoblot patterns. It appeared that some of the secreted factor XII Tenri formed several types of disulfide-linked complexes, including a factor XII-alpha1-microglobulin complex, through a newly generated Cys residue. The monomeric form of factor XII Tenri, like normal factor XII, was degraded into 2 major fragments with molecular weights of approximately 45 kDa and 30 kDa following mixing with activated partial-thromboplastin-time measuring reagent (cephalin and ellagic acid), whereas the factor XII Tenri that formed the complexes was not. This indicates that the factor XII Tenri present in disulfide-linked complexes with other proteins (and itself) is not converted to active forms, suggesting that attached proteins obstruct or delay the activation of factor XII via an inhibition of its binding to a negatively charged surface in vitro.     

Malignant Cystosarcoma Phyllodes Of Prostate

A case of malignant cystosarcoma phyllodes of the prostate is reported in a 45-year-old male. This tumor was composed of benign columnar or squamous cystic folds and sarcomatous stroma including rhabdomyomatous elements. The prostatic origin of the tumor was clearly proved by the unlabeled immunoperoxidase method. ACTA PATHOL. JPN. 34: 663–668, 1984.     

Effects of poly(ethyleneglycol)-modified hemoglobin vesicles on agonist-induced platelet aggregation and RANTES release in vitro

We studied the effects of hemoglobin-vesicles modified with PEG (PEG-HbV), a type of liposome-encapsulated hemoglobin (LEH), on human platelet functions in vitro. The effect of a low concentration of PEG-HbV (Hb; 5.8 mg/dl) was assessed by examining an agonist-induced aggregation response, and that of relatively high concentrations of PEG-HbV (Hb; 0.29, 1 and 2 g/dl) by measuring the release of RANTES (Regulated upon activation, normal T-cell expressed and presumably secreted) from platelets, which is regarded as a marker of platelet activation. The preincubation of platelets with PEG-HbV at 5.8 mg/dl of Hb did not affect platelet aggregation induced by collagen, thrombin and ristocetin. The pretreatment of platelet-rich plasma (PRP) with PEG-HbV at concen trations up to 2 g/dl of Hb had no aberrant effects on the collagen-induced RANTES release. Furthermore, the collagen-induced release of RANTES from PRP was not affected by longer incubation with PEG-HbV at 2 g/dl of Hb. The basal levels of RANTES from PRP were unchanged in the presence of PEG-HbV. These results suggest that PEG-HbV, at the concentrations studied, have no aberrant effects on platelet functions in the presence of plasma.     

Wnt signaling plays an essential role in neuronal specification of the dorsal spinal cord

In the developing spinal cord, signals from the roof plate are required for the development of three classes of dorsal interneuron: D1, D2, and D3, listed from dorsal to ventral. Here, we demonstrate that absence of Wnt1 and Wnt3a, normally expressed in the roof plate, leads to diminished development of D1 and D2 neurons and a compensatory increase in D3 neuron populations. This occurs without significantly altered expression of BMP and related genes in the roof plate. Moreover, Wnt3a protein induces expression of D1 and D2 markers in the isolated medial region of the chick neural plate, and Noggin does not interfere with this induction. Thus, Wnt signaling plays a critical role in the specification of cell types for dorsal interneurons.     

Proliferation of exogenously injected primordial germcells （PGCs） into busulfan-treated chicken embryos

Aim: This study was designed to investigate the effect of busulfan treatment on the proliferation of chicken primordialgerm cells (Fgcs) in vivo, focusing on the preferential settlement of PGCs onto the germinal ridges of chicken em-bryos. Methods: Bustdfan (250 rig/egg) was injected into the egg white of freshly oviposited fertilized eggs, whichwere then incubated. Embryonic developnent and viability were examined, and exogenous PGCs collected from embry-onic blood vessels were injected into the germinal crescent region of recipient enthryos. The nttmber of PGCs residedonto germinal ridges of the right and left sides were compared. Results: Bustdfan had a slight harmful effect on theembryo viabihty and the PGCs proliferation. The number of PGCs resided onto the left side of germinal ridges wasslightly higher as compared with the right side. Conclusion: Busulfan suppressed the viability of embryos and the pro-liferation of endogenous PGCs in the recipient embryos. However, the number of exogenous PGCs proliferated washigher in embryos treated with busnlfan than those without busulfan. Data also suggest the possibihty of a preferentialresidence of PCCs toward the left side of the germinal crescent region as compared with the right, which may be due toa more advanced functional development of the left gonad than the right. (Asian JAndro11999 Dec; 1 : 187 - 190)     

Reproductive characteristics of transgenic (TG) chickens carrying an exogenous gene

An exogenous gene (lacZ/MiwZ) introduced into the germinal crescent region (GCR) of avian embryos was confirmed to be successfully transferred to the gonads via the primordial germ cells (PGCs). Following hatching, the chicks were raised until the stage of sexual maturation. The incorporation of MiwZ DNA was detected in male and female transgenic chickens, respectively. The normal male and female transgenic birds were subjected to artificial insemination according to routine methods. Fertilized eggs obtained from female transgenic chickens were incubated for 72 h and the embryos removed from the yolk were examined by X-gal staining to detect the introduction of MiwZ in the offspring. As a result, the expression of MiwZ was detected in the offspring. Furthermore, the presence of MiwZ in the extracts from embryos was also detected by polymerase chain reaction (PCR) analysis. In male transgenic chickens, the presence of injected MiwZ in the extracts from sperm was also confirmed. The exogenous gene introduced into the GCR migrated successfully to the gonad resulting in its incorporation into the offspring and spermatozoa of transgenic chickens.     

Optic-spinal form of multiple sclerosis and immune-mediated myelopathy in Japan

Optic-spinal form of multiple sclerosis (OS-MS) and HTLV-I associated myelopathy/tropical spastic paraparesis (HAM/TSP) are two immune-mediated myelopathy relatively common in Japan. (1) Transverse myelitis, once seen in 60% of MS, mostly OS-MS, 30 years ago, drastically decreased (5%) recently in Japan. In contrast, frequency of conventional form of MS (C-MS) increased during this period of time. But unlike C-MS in white patients, cerebellar hemispheric lesions are uncommon in Japanese C-MS. These findings emphasize influence of changes in exogenous factors on manifestations of MS and distinct genetic factors related to MS in Japanese and white patients. (2) To clarify the reason of high HTLV-I proviral load in HAM/TSP, we studied cellular immune surveillance against HTLV-I and found that significant cytotoxic T lymphocyte activity, and suppressed natural killer activity and antibody-dependent cell-mediated cytotoxicity in the patients. These altered immune surveillance may be associated with the spread of HTLV-I infection and the pathogenesis of HAM/TSP.