Properties of human tissue isoferritins.

1. Human liver ferritin was separated by preparative isoelectric focusing into six fractions. 2. Except for the least acidic fraction the reactivity with antibody against spleen ferritin increased with rising pI, but with antibody against heart ferritin the reactivity decreased. 3. The highest iron content was found in the most acidic isoferritins and progressively decreased with rising pI. 4. Iron uptake was studied in apoferritin prepared from heart and liver ferritin fractions separated by ion-exchange chromatography. There was good correlation between the rate of iron uptake and pI. The most acidic fractions took up iron more rapidly than did the more basic ones. 5. Ferritin was prepared from heart, liver, spleen and kidney. There was little difference on isoelectric focusing between ferritin obtained from normal tissues and the corresponding iron-loaded tissues from patients who had received multiple blood transfusions. The iron-loaked heart ferritin invariably contained relatively more of the basic isoferritins. Normal and iron-overloaded heart ferritins were separated into isoferritin fractions by ion-exchange chromatography, and in each case there was a fall in iron content as the pI increased. The iron content of ferritin from the iron-overloaded heart was higher throughout than that from normal heart. 6. There is a relationship between the rate of iron uptake by apoferritin and pI, and this probably accounts for the variation in iron content of the isoferritins found in human liver and heart.     

Unknown New Zealand angiosperm assigned toCunoniaceae using sequence of the chloroplastrbcL gene

We report the first example of identification of an unknown flowering plant to family level using comparison of DNA sequences. The newly-discovered New Zealand plant had not been identified by traditional methods, but has been placed in the Gondwanic familyCunoniaceae on the basis of the sequence of its chloroplastrbcL gene. It has not flowered, but its vegetative characters are typical of the family. However, it is clearly unusual in several morphological and anatomical features and appears to match no known genus. We consider that, although it probably represents an unnamed new genus, it could possibly be a mutant dwarf form ofWeinmannia racemosa, so it is not prudent to name it at this time.     

Decay of the water reed Phragmites communis caused by the white-rot fungus Phlebia tremellosa and the influence of some environmental factors

Applied Microbiology and Biotechnology - The strain Phlebia tremellosa SBUG 1630 isolated from a thatched roof in Northern Germany is capable of colonizing and degrading effectively the water reed...     

Treatment with tumour infiltrating lymphocytes and interleukin-2 in patients with metastatic melanoma: A pilot study

Tumour infiltrating lymphocytes (TIL) were isolated and expanded from biopsy samples of 4 patients with metastatic melanoma. The patients were treated with autologous expanded TIL and continuous or bolus infusion of Interleukin 2 (IL-2) at a dose of 18 × 10⁶ International Units/m²/day for 5 days starting 36–48 hours after administration of cyclophosphamide at a dose of 1 g/m². The number of TIL infused ranged from 10¹⁰ to 5,56 × 10¹⁰ cells. Two patients had stable disease (SD) lasting for 2 1/2 and 4 months respectively and they died 24 and 13 months after therapy. One patient died during therapy due to a pseudomonas septicaemia and another patient developed progressive disease (PD). He died 3 months after the start of therapy. The side effects were substantial but most of them were reversible upon cessation of the treatment.The majority of the expanded TIL of all patients were of the CD8+ phenotype. Cutaneous metastases from two patients, removed after treatment with IL-2 and TIL, showed moderate lymphocytic infiltration also mainly of CD8+ T cells.The treatment with IL-2 and TIL is feasible, but further investigations should continue in an attempt to improve the efficacy of the therapy, to reduce toxicity and to diminish the costs and labour of the culture methods.     

The topology,structure and PE interaction of LITAF underpin a Charcot-Marie-Tooth disease type 1C

Background

Mutations in Lipopolysaccharide-induced tumour necrosis factor-α factor (LITAF) cause the autosomal dominant inherited peripheral neuropathy, Charcot-Marie-Tooth disease type 1C (CMT1C). LITAF encodes a 17 kDa protein containing an N-terminal proline-rich region followed by an evolutionarily-conserved C-terminal ‘LITAF domain’, which contains all reported CMT1C-associated pathogenic mutations.

Results

Here, we report the first structural characterisation of LITAF using biochemical, cell biological, biophysical and NMR spectroscopic approaches. Our structural model demonstrates that LITAF is a monotopic zinc-binding membrane protein that embeds into intracellular membranes via a predicted hydrophobic, in-plane, helical anchor located within the LITAF domain. We show that specific residues within the LITAF domain interact with phosphoethanolamine (PE) head groups, and that the introduction of the V144M CMT1C-associated pathogenic mutation leads to protein aggregation in the presence of PE.

Conclusions

In addition to the structural characterisation of LITAF, these data lead us to propose that an aberrant LITAF-PE interaction on the surface of intracellular membranes contributes to the molecular pathogenesis that underlies this currently incurable disease.

     

Phylogenetic relationships of New Zealand Asteraceae inferred from ITS sequences

Forty-five sequences from members of all genera of Asteraceae indigenous to New Zealand and 50 published sequences representing the tribal diversity in the family were analyzed to assess the utility of ITS sequences to resolve phylogenetic relationships. Previous studies using chloroplast DNA sequences and morphology provided support for several clades in the Asteraceae, yet the relationships among some of these were uncertain. The results from ITS analysis were largely consistent with these earlier studies. The New Zealand species are included in at least six clades, most of these corresponding to recognized tribes. Our results have also clarified the tribal affinities of a few anomalous genera. Haastia, previously aligned with the Gnaphalieae or the Astereae, is nested in the Senecioneae. Centipeda, previously included in the Astereae or Anthemideae, emerges near the Heliantheae. The relationships of Abrotanella remain unresolved. Received August 8, 2001 Accepted November 6, 2001     

Nucleocytoplasmic shuttling of the Duchenne muscular dystrophy gene product dystrophin Dp71d is dependent on the importin α/β and CRM1 nuclear transporters and microtubule motor dynein

Even though the Duchenne muscular dystrophy (DMD) gene product Dystrophin Dp71d is involved in various key cellular processes through its role as a scaffold for structural and signalling proteins at the plasma membrane as well as the nuclear envelope, its subcellular trafficking is poorly understood. Here we map the nuclear import and export signals of Dp71d by truncation and point mutant analysis, showing for the first time that Dp71d shuttles between the nucleus and cytoplasm mediated by the conventional nuclear transporters, importin (IMP) α/β and the exportin CRM1. Binding was confirmed in cells using pull-downs, while in vitro binding assays showed direct, high affinity (apparent dissociation coefficient of c. 0.25 nM) binding of Dp71d to IMPα/β. Interestingly, treatment of cells with the microtubule depolymerizing reagent nocodazole or the dynein inhibitor EHNA both decreased Dp71d nuclear localization, implying that Dp71d nuclear import may be facilitated by microtubules and the motor protein dynein. The role of Dp71d in the nucleus appears to relate in part to interaction with the nuclear envelope protein emerin, and maintenance of the integrity of the nuclear architecture. The clear implication is that Dp71d's previously unrecognised nuclear transport properties likely contribute to various, important physiological roles.     

Novel mechanisms of biotransformation of p-tert-amylphenol by bacteria and fungi with special degradation abilities and simultaneous detoxification of the disinfectant

The compound p-tert-amylphenol (p-(1,1-dimethylpropyl)phenol) is a widely used disinfectant belonging to the group of short branched-chain alkylphenols. It is produced in or imported into the USA with more than one million pounds per year and can be found in the environment in surface water, sediments, and soil. We have investigated for the first time the biotransformation of this disinfectant and the accumulation of metabolites by five bacterial strains, three yeast strains, and three filamentous fungi, selected because of their ability to transform either aromatic or branched-chain compounds. Of the 11 microorganisms tested, one yeast strain and three bacteria could not transform the disinfectant despite of a very low concentration applied (0.005 %). None of the other seven organisms was able to degrade the short branched alkyl chain of p-tert-amylphenol. However, two yeast strains, two filamentous fungi, and two bacterial strains attacked the aromatic ring system of the disinfectant via the hydroxylated intermediate 4-(1,1-dimethyl-propyl)-benzene-1,2-diol resulting in two hitherto unknown ring fission products with pyran and furan structures, 4-(1,1-dimethyl-propyl)-6-oxo-6-H-pyran-2-carboxylic acid and 2-[3-(1,1-dimethyl-propyl)-5-oxo-2H-furan-2-yl]acetic acid. While the disinfectant was toxic to the organisms applied, one of the ring cleavage products was not. Thus, a detoxification of the disinfectant was achieved by ring cleavage. Furthermore, one filamentous fungus formed sugar conjugates with p-tert-amylphenol as another mechanism of detoxification of toxic environmental pollutants. With this work, we can also contribute to the allocation of unknown chemical compounds within environmental samples to their parent compounds.