Nonsense mutations of the von Willebrand factor gene in patients with von Willebrand disease type III and type I.

von Willebrand disease (vWD) is the most common inherited bleeding disorder in humans. The disease is caused by qualitative and quantitative abnormalities of the von Willebrand factor (vWF). Genomic DNA from 25 patients with vWD type III, the most severe form of the disease, was studied using PCR followed by restriction-enzyme analysis and direct sequencing of the products. Nonsense mutations (CGA----TGA) were detected in exons 28, 32, and 45 by screening of all the 11 CGA arginine codons of the vWF gene. Two patients were found to be homozygous and five heterozygous for the mutation. Both parents and some of the relatives of the homozygous patients carry the mutation. These are the first reported examples of homozygous point mutations associated with the severe form of vWD. In the three heterozygous probands, one of the parents carried the mutation and had vWD type I. Family studies including parents and family members with or without vWD type I indicated that these three heterozygous patients are likely to be compound heterozygous. Twenty-one individuals from these seven families with vWD type I were found to be heterozygous for the mutation.     

Generation of hydroxyl radical by crocidolite asbestos is proportional to surface [Fe3+].

Differences among fibrous silicates to effect injury in biological systems have been postulated to reflect oxidant generation by structural iron within the crystal lattice of amphiboles. Iron is also coordinated to the surface of all silicates in concentrations which depend on the density of acidic functional groups. We tested the hypothesis that oxidant generation by crocidolite is proportional to surface-complexed iron rather than variance in the lattice concentrations of this transition metal. Surface iron was quantified after its reduction to Fe2+ and chelation by citrate. Thiobarbituric acid (TBA) reactive products and dihydroxybenzoic acid products of salicylate were employed as indices of nonspecific oxidant and hydroxyl radical generation, respectively. Surface iron, TBA reactive products, and dihydroxybenzoic acid products all diminished after pretreatment of crocidolite with the metal chelator deferoxamine in concentrations varying from 0 to 250 mM. Inclusion of deferoxamine in the reaction mixture provided similar results of diminishing both TBA reactive products and dihydroxybenzoic acid generation. We conclude that oxidant generation by crocidolite is proportional to surface concentrations of iron which can be chelated using deferoxamine. The design of synthetic fibers without health effects after exposure will likely necessitate decreasing the number of surface acidic functional groups to diminish the capacity to complex iron (i.e., minimize the percentage SiO2).     

Uptake and metabolism of N-(4'-pyridoxyl)amines by isolated rat liver cells.

Uptake and metabolism of [3H]pyridoxine and 3H-labeled N-(4'-pyridoxyl)amines by isolated rat liver cells were studied at physiological concentration (0.5 microM) of vitamin B6 by using both membrane filtration and centrifugation methods for removal of radiolabeled solutes after incubations with cells. It was found that the characteristics of import of N-(4'-pyridoxyl)amines into liver cells is similar to those of import of natural vitamin B6. Upon entry each 4'(N)-substituted pyridoxamine was converted to its 5'-phosphate and then oxidized to release pyridoxal 5'-phosphate and the original amine. Considerable size of the amine substituent is tolerated for transport and metabolism, but a charged function impedes entry. The amount of released pyridoxal 5'-phosphate (and therefore the amount of released original amine) is controlled partially by the size of the amine affixed to B6 and partially by the enzymatic steps involved. This system illustrates how biologically active amines can be piggybacked onto a vitamin that gains facilitated entry to cells that have the enzymatic means to release the free amine for subsequent effects within the cell.     

Hepatitis B virus DNA integration and expression of an erb B-like gene in human hepatocellular carcinoma.

Southern blot studies on Hepatitis B Virus (HBV) DNA integration in 13 human hepatocellular carcinomas (HCCs) patients revealed the presence of several distinct HBV integration sites in different human liver disease patients. In one HCC patient the DNA fragment containing the HBV integration also hybridized to an erb B probe. The erb B/HBV co-migrating DNA fragment was cloned and sequenced, and showed that HBV DNA is integrated next to a cellular DNA fragment which is homologous to the tyrosine protein kinase domain of the human epidermal growth factor receptor gene and other cell surface receptor genes. The virus-integrated cellular DNA sequence is expressed in this HCC patient, suggesting a possible role for this gene in hepatocarcinogenesis.     

Parametric study of diethyl phthalate biodegradation

Summary The effects of temperature, dissolved oxygen, and other environmental parameters under both aerobic and anaerobic conditions were investigated using one aerobic and one facultative strain isolated from wastewater treatment plant sludge. Among other results, we found that low dissolved oxygen levels and low temperatures decreased the rate of DEP degradation and the growth rate, and that the facultative strain was much less affected by the lower DO concentrations than the aerobic strain.     

Mitochondrial gene mutation: the ageing process and degenerative diseases

Polymerase chain reaction (PCR) amplification was carried out on total DNA from a range of autopsy tissues from deceased human subjects with no known mitochondrial disease, aged from birth (80 minutes) to 87 years. We report the finding of an age-related 5 kb deletion in the mitochondrial genomes of these subjects. The deletion occurs between nucleotide positions 8470 and 13459 of the mitochondrial genome, and is flanked by a 13 bp direct repeat. All tissues from adult subjects showed the presence of mitochondrial DNA molecules with the deletion after a 30 cycle PCR amplification; by contrast the deletion was not similarly detected in any of the infant tissues analysed. However, the occurrence of the deletion was detected in the infant tissues after 60 PCR cycles of MtDNA amplification. It is concluded that such deletions are not necessarily associated with particular mitochondrial diseases but occur naturally, and with increasing frequency with age. A consequence of the accumulation of this deletion could be a progressive decrease with age of bioenergetic capacity which in turn could influence the rate of ageing and predispose to age-associated degenerative diseases.     

L7811鼠腹水肿瘤细胞^31P核磁共振的研究

用~(31)P核磁共振技术(~(31)P-NMR)研究了L_(7811)鼠腹水肿瘤细胞和615系鼠胸腺细胞(正常对照细胞)。结果发现在肿瘤晚期阶段,L_(7811)腹水肿瘤细胞的含磷化合物未进入完全不活跃状态。此外,腹水肿瘤细胞的磷脂组成与含量亦有明显改变。因此,~(31)P-NMR谱可做为观察肿瘤细胞内能量生成和某些磷脂合成宏观动态过程的一项参考指标。     

急性缺氧下听觉运动反应的研究

受试者为9名男性青年。在低压舱模拟海拔3000、4000、5000和6000m,分别进行1h实验,共36人次。各高度上停留时作听觉检测,并记录了脑电和主诉。结果表明,在3000m,人的听觉运动反应基本正常;而在5000m、6000m,听觉运动反应的正确率、延时率、遗漏率和反应时等多项指标发生显著变化,工效降低。本研究为急性缺氧防护生理标准和缺氧耐力提供了工效的依据。