Can EGFR mutation status be reliably determined in pre‐operative needle biopsies from adenocarcinomas of the lung?

The identification of EGFR mutations in non‐small‐cell lung cancer is important for selecting patients, who may benefit from treatment with EGFR tyrosine kinase inhibitors. The analysis is usually performed on cytological aspirates and/or histological needle biopsies, representing a small fraction of the tumour volume. The aim of the present investigation was to evaluate the diagnostic performance of this molecular test. We retrospectively included 201 patients with primary adenocarcinoma of the lung. EGFR mutation status (exon 19 deletions and exon 21 L858R point mutation) was evaluated on both pre‐operative biopsies (131 histological and 70 cytological) and on the surgical specimens, using PCR. Samples with low tumour cell fraction were assigned to laser micro‐dissection (LMD). We found nine (4.5%) patients with EGFR mutation in the lung tumour resections, but failed to identify mutation in one of the corresponding pre‐operative, cytological specimens. Several (18.4%) analyses of the pre‐operative biopsies were inconclusive, especially in case of biopsies undergoing LMD and regarding exon 21 analysis. Discrepancy of mutation status in one patient may reflect intra‐tumoural heterogeneity or technical issues. Moreover, several inconclusive results in the diagnostic biopsies reveal that attention must be paid on the suitability of pre‐operative biopsies for EGFR mutation analysis.     

Increase of nucleoid size with increasing age of hyphal region in vegetative mycelia of Streptomyces granaticolor

In mycelia of Streptomyces granaticolor grown in liquid cultures the distribution of DNA was surveyed by staining and autoradiographic methods. The frequency of stained nucleoids was rather similar in young and old hyphal regions. However, the size of the nucleoids increased with the age of a hyphal region. The degree of heterogeneity in nucleoid size was particularly high during slow growth. In the autoradiograms silver grains (³H-thymidine label) were also heterogeneously distributed along the hyphae. Calculation of the average number of silver grains per nucleoid revealed that the amount of DNA per nucleoid was 1.6 times higher in the more distant subapical regions than within the 35 μm long apical region. From a chase experiment it became obvious that the DNA segregated in a normal manner only within the apical region. When the chase lasted for one doubling time, the frequency of silver grains was reduced to 50% only within the about 30 μm long apical region. The label incorporated subapieally remained almost constant, therefore, indicating a lack of normal segregation. Only about 10% of the label was lost by branching. The lack of segregation of sister nucleoids after repeated rounds of replication was found to be responsible for the increase of nucleoid size within subapieal hyphal regions.     

Constitutive and 3-methylcholanthrene-induced rat ALDH3A1 expression is mediated by multiple xenobiotic response elements.

The rat class 3 aldehyde dehydrogenase gene (ALDH3A1) is expressed constitutively or by xenobiotic induction depending on the tissue in which it occurs. Although the mechanism that mediates inducible expression has been well characterized, relatively little is known about constitutive regulatory mechanisms. Previous ALDH3A1 promoter analyses have indicated that primary regulatory regions within the ALDH3A1 5' flanking region exert similar effects on both constitutive and inducible ALDH3A1 expression. However, promoter gene analyses that served as the basis of early work were limited by the lack of sufficient 5' flanking region sequence. To gain a more complete picture of how the 5' flanking region regulates both modes of expression, we have subcloned an 8.0-kilobase (kb) fragment from the 5' flanking region of the ALDH3A1 gene and subjected it to reporter gene analyses. We found a region located between 4.8 and 7.8 kb upstream of the noncoding first exon that drives strong ALDH3A1 reporter activity. This region contains xenobiotic response element consensus sequences that mediate constitutive and inducible ALDH3A1 reporter gene expression. Using the new generation of ALDH3A1 reporter constructs, we were unable to confirm the presence of a negative regulatory region that was apparent in previous studies using a shorter fragment of the 5' flanking region. We also demonstrate that 3-methylcholanthrene induces ALDH3A1 expression above high constitutive background in corneal epithelial cells.     

Die Entwickelung der Vorderen Augenkammer

Ohne Zusammenfassung     

Metabolism of aniline and 3-chloroaniline in Pseudomonas acidovorans CA28: Evidence of isofunctional muconate cycloisomerases

Two distinct muconate cycloisomerases are involved in the degradation of aniline and 3-chloroaniline (3-CA) proceeding via the ortho-cleavage pathways in Pseudomonas acidovorans CA28. After partial purification of these two enzymes kinetic investigations resulted in a clear differentiation of aniline-derived muconate cycloisomerase (MC) and 3-CA-derived chloromuconate cycloisomerase (CMC). A further result of this study revealed the simultaneous coexistence of MC and CMC in strain CA28 when grown on a mixture of aniline and 3-CA.     

Effect of whole saliva on the rheologic behavior of extracellular water-soluble glucan produced by Streptococcus mutans.

The viscosity of mixtures of Streptococcus mutans water-soluble glucan and stimulated whole saliva or buffer was measured at pH 5, 6, 7, and 8. The viscosity was measured as a function of shear rate in the range 15 s-1-230 s-1. Though the centrifuged saliva had a viscosity close to that of water it increased the viscosity of the glucan up to 65% at pH 6 and 55% at pH 7 and at a shear rate of 20 s-1, indicating an interaction between saliva components and glucan that could be an important part of the cohesive forces of plaque matrix. The interaction between saliva and glucan was less pronounced at pH 5 and 8, which indicates a charge-dependent interaction. The viscosity increase at pH 6 and 7 was higher at low than at high shear rates, suggesting a higher contribution to plaque stability when weak as opposed to high mechanical forces are exerted on the plaque.