Arthrogryposis,renal dysfunction and cholestasis syndrome: Report of five patients from three Italian families

We report on five patients from three families with neurogenic arthrogryposis, cholestasis and tubular renal dysfunction. Despite a similar clinical picture the liver histology showed a broad pathological spectrum, ranging from pigment storage to parenchymal giant cell transformation and ductopenia. The findings are compared with those of other cases from the literature in search of a correct nosology of the syndrome characterized by arthrogryposis, renal and liver disease.Conclusion We propose to consider the picture of arthrogryposis, renal tubular dysfunction and cholestasis as a single syndrome.     

A novel strategy for the isolation of defined pyrG mutants and the development of a site-specific integration system for Aspergillus awamori

A homologous gene transfer system for Aspergillus awamori for site-specific integration is described, based on two components. First, a defined A. awamori pyrG mutant strain constructed by a selection strategy for gene-replacement in fungi. Second, a vector with a homologous pyrG selection marker containing a defined mutation at a site different from that of the mutations in the pyrG gene of the defined mutant strain. Defined mutation in the A. awamori pyrG gene, isolated from a genomic library by heterologous hybridisation with the A. niger pyrG gene as a probe, were introduced by specifically altering sequences at restriction sites in the coding region of the gene. After transformation of the A. awamori wild-type strain with vectors containing these mutated pyrG genes, and selection for 5-fluoro-orotic acid resistance (5-FOA^R), on the average 60% of the 5-FOA^R colonies originated from replacement of the wild-type pyrG gene by the mutated pyrG allele. After transformation of a mutant strain, carrying a mutation near the 5 end of the pyrG gene with vectors containing a mutation near the 3 end of the pyrG gene, 35% of the resulting transformants contained one copy of the vector at the pyrG locus.     

DNA polymerases α, δ, and ɛ of Novikoff hepatoma cells differ from those of normal rat liver in physicochemical and catalytic properties

To investigate whether DNA replication in malignant cells deviates from that of normal cells we compared DNA polymerases , , and from normal rat liver to the enzymes from fast-growing (malignant) Novikoff hepatoma cells. DNA polymerases were purified 300-fold by three chromatographic steps. Characterization included measurement of physicochemical constants (including sedimentation coefficients, diffusion coefficients, calculation of relative molecular masses), quantitation of catalytic activities using specific DNA primer templates (K _m values) and inhibitors (K _i values), and identification of polypeptides which are strongly associated with DNA polymerases. Comparison of physicochemical and catalytic properties of DNA polymerases from both sources revealed similarities but also some important differences. DNA primase associated with DNA polymerase , and 3–5 exonuclease accompanying DNA polymerases and had similar activities. In contrast, the DNA-binding domain of DNA polymerases and from hepatoma cells was altered since K _m values, determined with the specific primer templates gapped calf thymus DNA and poly(dA·dT), were higher. Furthermore, sedimentation and diffusion coefficients, Stokes' radii, and frictional coefficient ratios of DNA polymerases and from malignant cells significantly deviated. In addition, when the dNTP-binding sites were probed with specific inhibitors (aphidicolin, butylphenyl-dGTP, carbonyldiphosphonate, and dideoxy-TTP), significantly lower K _i values were obtained for the polymerases from Novikoff cells indicating lower affinity of the dNTP binding site to deoxyribonucleoside 5-triphosphates. Altered catalytic and molecular properties are possibly a consequence of malignant transformation. It is to be expected that similar changes occur in DNA polymerases of other tumors. In particular, diminished affinity to primer templates and weakened nucleotide binding leads to lowered specificity of nucleotide selection in the base-pairing process and is therefore likely to cause an enhanced mutation rate during malignant progression.Abbreviations PCNA 3 Proliferating-cell nuclear antigen This paper is dedicated to Prof. Dr. R. Neidlein on the occasion of his 65th birthday.     

当归补血汤对细胞免疫调节作用及超微结构的影响

本实验采用环磷酰胺(CY)制造免疫功能低下小鼠的动物模型,研究当归补血汤组成药物的不同比例配伍,对机体免疫功能调节作用的影响。实验过程中将当归、黄芪分别配制成1∶1(A方)、1∶5(B方)、5∶1(C方)的不同比例中药方剂,对小鼠施以灌胃,并设对照组,以WBC总数、T淋巴细胞转化率及胸腺细胞超微结构变化作为检测指标。结果表明:三组不同比例配伍的方剂,对小鼠机体免疫功能均有调节作用。三组之间比较,提高WBC总数以A方效果最明显;三个方剂均有促进淋巴细胞转化的作用,但转化率三者之间无明显差异;保护胸腺细胞作用以B方效果最好。应用:当归补血汤对机体免疫功能有明显调节作用,但不同比例配伍,对机体免疫功能调节作用各有所侧重     

Blastic transformation of mantle cell lymphoma: genetic evidence for a clonal link between the two stages of the tumour

AIMS: The blastic variant of mantle cell lymphoma (MCL-BV) may develop through histological transformation of mantle cell lymphoma (MCL). However, the clonal link between the tumour cells of MCL and transformed MCL-BV has not been established at the genetic level. To investigate this link longitudinal molecular genetic studies have been performed in two cases of MCL that showed morphological transformation to MCL-BV. METHODS AND RESULTS: Polymerase chain reaction (PCR) and nucleotide sequence analyses of the complementary determining region 3 (CDR) of the immunoglobulin (Ig) heavy chain (H) gene were performed to identify clone-specific rearrangements. In both cases, nucleotide sequence analysis revealed common clone-specific IgH gene rearrangements in MCL and subsequent MCL-BV. CONCLUSIONS: These results provide genetic evidence for the common clonal origin of MCL and subsequently developed MCL-BV.     

In vitro and in vivo uptake of nickel sulfides by rat lymphocytes

The uptake, the biological transformation and the interaction with cellular constituents of Ni₃S₂ and NiS have been studied in vitro and in vivo on rat lymphocytes. NiS crystals are phagocytized in vitro and no structural degradation is observed within the first 3 days of exposure. Energy dispersive spectrometry (EDS) reveals a slight dissolution characterized by the loss of sulfur. Ni₃S₂ is degraded in the extracellular space to minute particles (50–100 nm) covering the cell membrane. Smaller intracellular particles (10–30 nm) are found selectively bound to mitochondria, endoplasmic reticulum, Golgi vesicles, nuclear membranes, and the euchromatinic part of nuclei. EDS analyses reveal that the particles bound to cell membranes and euchromatin no longer contain sulfur but phosphorus and nickel as inorganic compounds. This observation suggests the formation of a Ni/P complex with the phosphate groups either of membranous phospholipids or of nuclear RNA or DNA. A similar uptake and transformation process of Ni₃S₂ is observed on lymphocytes after in vivo incubation. This leads us to consider lymphocytes as target cells, as compared with other cell types where the Ni₃S₂ uptake occurs only partially. The present findings show a difference of uptake and biological transformation between Ni₃S₂ and NiS. The identical results obtained after in vitro and in vivo bioassays enhance the in vitro experiments, at least for this cell type.     

Steady-state kinetics of imipramine in patients

Steady-state plasma level kinetics were studied in 76 patients given imipramine (IP) 150 to 225 mg/day for 2–5 weeks. IP was given in three divided doses at 8.00 a.m., 1.00 p.m. and 5.00 p.m. Plasma concentrations of IP and its active metabolite desipramine (DMI) were determined by quantitative in situ thin-layer chromatography. The plasma levels of IP and DMI showed pronounced flucutations throughout the day with a ratio of about 2 between highest and lowest level. Patients with steady-state levels of IP and/or DMI below 50 g/l reached this within 1 week of treatment. Patients with higher steady-state levels reached steady-state concentrations within 2–3 weeks. There were some intraindividual fluctuations in plasma levels from week to week after steady state had been reached (coefficient of variation: 10–20%). Interindividually, the steady-state levels corrected to a dose of 3.5 mg/kg per day varied considerably: IP: 6–356 g/l, DMI: 24–659 g/l and IP+DMI: 58–809 g/l. The steady-state plasma levels showed a skew distribution that became normal by logarithmic transformation. The IP/DMI ratio ranged from 0.07 to 5.5 with a median value of 0.47. Compared to data from amitriptyline treated patients the IP/DMI ratios had significantly lower median value and larger variation than the corresponding plasma level ratios of amitriptyline/nortriptyline. Several statistically significant differences in steady-state levels between age groups were found. For IP: Women aged 30–39 had lower levels than women aged 20–29, 40–49, and 50–59, and men aged 50–59 and 60–65; men aged 30–39 had lower levels than men aged 60–65. For DMI: Women aged 30–39 had lower levels than women aged 50–59.     

桔青霉对头孢呋辛酯的代谢转化

利用微生物转化的方法，选取桔青霉(Penicillium citrinum)为转化菌株，对头孢呋辛酯转化产率等方面进行了研究。采用枯草芽孢杆菌[CMCC(B)63501]作为检测菌株检测转化产物，结果表明，桔青霉能产生稳定的具抗菌活性的转化产物，且重现性好，转化产物的平均产率71．9％。高效液相色谱检测证明转化产物为头孢呋辛。     

根癌农杆菌对云南红豆杉原生质体的转化

本文介绍了根癌农杆菌T—DNA对云南红豆杉原生质体的转化，分析了转化系的紫杉醇合成能力。实验以生长20d的云南红豆杉幼茎诱导的淡黄色愈伤组织为材料，经酶解可分离出大量有活力的原生质体。在由B5无机盐、KM有机成分、3．0mg／L2，4-D、0．1mg／LKT和0．45mol／L果糖组成的培养基中培养1周后，原生质体再生细胞持续分裂形成细胞团。根癌农杆菌对原生质体的转化能力与原生质体生长状态及菌株相关。虽然刚分离的或已再生细胞壁的原生质体不能被根癌农杆菌转化，但由10个以上细胞组成的原生质体克隆和根癌农杆菌B6S3菌株共培养后可实现T—DNA转化，高压纸电泳检测转化系具有冠瘿碱的合成，转化率约为5％。HPLC证实转化系具有合成紫杉醇的能力，但不同转化系中紫杉醇含量变异很大，最高含量为0．076％，是对照愈伤组织的6倍，表明T—DNA的插入改变了培养细胞对紫杉醇的合成。获得的高产转化系细胞生长比对照低，但继代培养中其生长和紫杉醇积累基本保持稳定。尽管转化系合成紫杉醇能力远未达到商业化生产的要求，但研究通过T—DNA对原生质体的转化而实现插入诱变，可以为分离高产紫杉醇优良细胞系提供单细胞筛选系统。     

盐酸麻黄碱转化为盐酸伪麻黄碱工艺研究

盐酸伪麻黄碱 ( 1 )与盐酸麻黄碱 ( 2 )是麻黄草的主要药用成份 ,两者是差向异构体。 2的应用较广 ,因而由 1转化为 2的工艺报道较多 [1,2 ] ,而 2转化为1的方法相对较少 [3 ]。自 1 992年我国卫生部正式批准 1供临床使用后 ,国内研制开发含有 1复方制剂品种达 60多个 [4]。美国 PDR收载的含 1的非处方药物已达 1 5 0种以上[5] 。随着对 1需求量的不断增大 ,由 2转化为 1的工艺研究已越来越受到重视。文献 [3 ]先将 2游离 ,经乙醚提取、乙酸酐转化 ,再用 4倍摩尔量的亚硫酰氯进行水解 ,操作繁琐 ,三废较多。我们直接用乙酸酐转化 ,1 mol/L…