Retention of bile salts in micellar electrokinetic chromatography: relation of capacity factor to octanol–water partition coefficient and critical micellar concentration

The capacity factors of 16 anionic cholates (from six bile salts, including their glyco- and tauro-conjugates) were determined in a micellar electrokinetic chromatography (MEKC) system consisting of buffer, pH 7.5 (phosphate–boric acid; 20 mmol/l) with 50 mmol/l sodium dodecyl sulfate (SDS) as micelle former and 10% acetonitrile as organic modifier. The capacity factors of the fully dissociated, negatively charged analytes (ranging between 0.2 and 60) were calculated from their mobilities, with a reference background electrolyte (BGE) without SDS representing “free” solution. For comparison, the capacity factors were derived for a second reference BGE where the SDS concentration (5 mmol/l) is close to the critical micellar concentration (CMC). The capacity factors are compared with the logarithm of the octanol–water partition coefficient, log P_OW, as measure for lipophilicity. Clear disagreement between these two parameters is found especially for epimeric cholates with the hydroxy group in position 7. In contrast, fair relation between the capacity factor of the analytes and their CMC is observed both depending strongly on the orientation of the OH groups, and tauro-conjugation as well. In this respect the retention behaviour of the bile salts in MEKC seems to reflect their role as detergents in living systems, and might serve as model parameter beyond lipophilicity.     

Alpha(v)beta(6) integrin-A marker for the malignant potential of epithelial ovarian cancer.

The mechanism(s) responsible for the progression of non-metastatic or borderline ovarian cancer to invasive Grade I/III ovarian cancer is still unknown. An epithelium-restricted integrin, alpha(v)beta(6), is present in malignant epithelia but not in normal epithelia. We studied the relative expression and distribution of alpha(v)beta(6) integrin in early and late-stage invasive (Grade I and Grade III) and non-invasive (benign and borderline) ovarian tumors of serous, mucinous, endometrioid, and clear-cell carcinoma subtypes, to assess its potential as a marker for epithelial ovarian cancer progression. Sixty-six specimens, including eight normal, 13 benign, 14 borderline, 13 Grade I, and 18 Grade III tumors were evaluated by immunohistochemistry (IHC) using a monoclonal antibody (MAb) against alpha(v)beta(6) integrin. Normal ovarian surface epithelium was negative for alpha(v)beta(6) integrin expression. All 45 carcinomas studied were positive, and the staining intensity significantly correlated with the grade of the tumor. The Grade III carcinomas of all types showed strong staining intensity. Only mucinous benign tissues were positive, and no reactivity was observed in benign serous neoplasms. On the basis of these observations, we hypothesize that the expression of alpha(v)beta(6) integrin is associated with epithelial ovarian cancer and that a gradual increase in the expression of the molecule may be a correlative index of the progression of this disease.     

THE GEOTROPIC REACTION OF RODLESS MICE IN LIGHT AND IN DARKNESS

"Black—short ears—kinky tail—rodless" mice, controlled by "pink eyed—dilute—brown" mice, were tested on an inclined plane in order to determine if they are photically sensitive, and, if so, to get a quantitative expression for their visual receptivity. Rodless and control animals were tested in the dark to obtain an expression for normal geotropic orientation. Light was then introduced to modify these reactions if possible. Under light, the controls failed to orient, whereas the rodless gave reactions almost identical with those in the dark. This test has failed in this experiment to suggest sight in the rodless mouse.     

Analysis of the Albino-Locus Region of the Mouse: IV. Characterization of 34 Deficiencies

Thirty-four independent nonviable c-locus mutations (types c^al, albino lethal and c^as, albino subvital), derived from radiation experiments, were tested for involvement of nearby markers tp, Mod-2, sh-1, and Hbb: 10, 22, and 2 involved, respectively, none of these markers, Mod-2 alone, and Mod-2 plus sh-1. When classified on this basis, as well as according to developmental stage at which homozygotes die, and by limited complementation results, the 34 independent mutations fell into 12 groups. From results of a full-scale complementation grid of all 435 possible crosses among 30 of the mutations, we were able to postulate an alignment of eight functional units by which the 12 groups fit a linear pattern. Abnormal phenotypes utilized in the complementation study were deaths at various stages of prenatal or postnatal development, body weight, and reduction or absence of various enzymes. Some of these phenotypes can be separated by complementation (e.g., there is no evidence that mitochondrial malic enzyme influences survival at any age); others cannot thus be separated (e.g., glucose-6-phosphatase deficiency and neonatal death).—We conclude that all of the nonviable albino mutations are deficiencies overlapping at c, and ranging in size from <2cM to 6-11 cM. The characterization of this array of deficiencies should provide useful tools for gene-dosage studies, recombinant-DNA fine-structure analyses, etc. Since many of the combinations of lethals produce viable albino animals that resemble the standard c/c type, we conclude (a) that the c locus contains no sites essential for survival, and (b) that viable nonalbino c-locus mutations (c^xv) are the result of mutations within the c cistron. Viable albinos (c^av, the majority of radiation-induced c-locus mutations) may be intracistronic mutations or very small deficiencies.     

The measurement of protein synthesis in biological systems

Attempts to quantitate metabolism in the lung and other tissues using radioactive precursors may be subject to significant errors arising from inappropriate assumptions regarding precursor metabolism, compartmentation and specific radioactivity. This article reviews the type and magnitude of error which may complicate such measurements by presenting specific data from experiments using radioactive amino acids to estimate the rate of protein synthesis. The applicability of these observations to other metabolic systems is discussed briefly in order to develop a more general awareness of the errors which may result from incomplete validation of experimental measurements using radioisotopes.     

The adult β-globin genes of the “single” type mouse C57BL

A set of four clones containing the two adult β-globin genes of the “single” type mouse C57BL/10 (genotype Hbb^s/Hbb^s) were isolated from a library of cloned restriction fragments. The two genes, designated βs and βt, were physically mapped onto a 32 kb segment of the chromosome carried by the four clones. βs and βt form a stable heteroduplex 1850 bp long, indicating that they are intact and conserved at this level of resolution throughout their length, including their intervening sequences. The βs gene is allelic with the βdmaj gene of the BALB/ c mouse (genotype Hbb^d/Hbb^d). These two alleles, as well as their surrounding sequences, are highly conserved. In contrast, heteroduplexes of βt with its BALB/c allele, βdmin, revealed three extensive but localized rearrangements. One region of non-homology falls within the large intervening sequence, IVS2. To the 5′ side of the βt/βdmin gene position two unequal substitutions were observed; each results in the net insertion of about 1000 bp into the Hbb^d chromosome. The β/βdmin gene position is bracketed by a 1450 bp inverted repeat. One of the 1000 bp substitutions maps within this inverted repeat.