The human ovarian teratocarcinoma cell line PA-1 demonstrates a single translocation: analysis with fluorescence in situ hybridization, spectral karyotyping, and bacterial artificial chromosome microarray

Cell lines derived from tumors contain numerous chromosomal aberrations and are the focus of study in tumor evolution. The ovarian teratocarcinoma cell line PA-1 demonstrates a single chromosomal aberration: a reciprocal t(15;20)(p11.2;q11.2). A complete molecular genetic analysis was undertaken to characterize this cell line. The PA-1 cell line was studied with fluorescence in situ hybridization (FISH), spectral karyotyping (SKY), bacterial artificial chromosome (BAC) microarray, and Western blotting. Amplification of 20q is frequently implicated in both breast and ovarian cancer; this region contains a number of oncogenes including MDM2, ZNF217, and the ovarian tumor marker WFDC2 (alias HE4). FISH revealed gene amplification of AIB1 (now known as NCOA3) but not STK15 (now known as AURKA). Immunoblot analysis demonstrated 3.6-fold overexpression of the AIB1 protein product, but no elevation of the STK15. BAC cancer gene microarray analysis showed gene amplification of > or =1.20 for five oncogenes. The presence of a consistent single change in PA-1, the t(15;20)(p11.2;q11.2), suggests that the aberration is significant with respect to the transformation status of the cell line. This translocation appears to cause overexpression of AIB1 (and perhaps other proteins), which may provide an immortalizing effect on this cell line.     

Enhanced molecular volume of conservatively pegylated Hb: (SP-PEG5K)6-HbA is non-hypertensive

Recent studies have suggested that the "pressor effect" of acellular Hb is a consequence of perturbation of the macro-and microcirculatory system in multiple ways, and that PEGylation is an effective approach for controlling the same. In an attempt to confirm this concept, a new and simple thiolation mediated, maleimide chemistry-based conservative PEGylation protocol has been developed to conjugate multiple copies of PEG-chains to Hb. This approach combines the high reactivity of maleimides towards thiols with the propensity of iminothiolane to derivatize the epsilon-amino groups of proteins into reactive thiol groups, with conservation of their positive charge. One of the PEGylated products, namely (SP-PEG5K)6-HbA, that carries on an average six copies of PEG5000 chains per Hb, is non-hypertensive in hamster top load and in rat 50% exchange transfusion models. This hexa-PEGylated-Hb has (i) a hydrodynamic volume corresponding to that of an oligomerized Hb of 256kDa, (ii) a molecular radius of approximately 6.8 nm, (iii) high oxygen affinity, (iv) lowered Bohr effect, and (v) increased viscosity and colloidal osmotic pressure. These properties of (SP-PEG5K)6-HbA are consistent with the emerging new paradigms for the design of Hb based oxygen carriers and confirm the concept that the "pressor effect" of Hb is a multifactorial event. The thiolation mediated maleimide chemistry-based PEGylation protocol described here for the generation of (SP-PEG5K)6-Hb is simple, highly efficient, and is carried out under oxy conditions. The results demonstrate that a non-hypertensive PEG-Hb can be generated by conjugation of a lower number of PEG chains than previously reported.     

Author index for volume 106, number 1

Treatment of mouse L929 cells with immune T (type II) interferon resulted in the induction of two double-stranded (ds) RNA-dependent enzymes which have been described previously in viral (type I) interferon-treated cells: pppA(2′p5′A)n synthetase and protein kinase(s). The induction of these enzymes by T interferon was blocked by anti-T but not by antiviral interferon serum. The kinetics of the development of the antiviral state along with a comparison of the levels of pppA(2′p5′A)n synthetase and protein kinase in T and viral interferon-treated cells were investigated. In addition, [³⁵S]methionine-labeled extracts from control, T, and viral inteferon-treated cells were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and two-dimensional gel electrophoresis. Extracts from cells treated with either type of interferon were found to contain several identical proteins, of estimated molecular weights 60,000, 88,000, and 120,000, which were absent or found at much lower levels in preparations from control cells. The newly synthesized proteins can be partially purified by chromatography on columns of poly(I)·poly(C)-Sepharose, thus providing a convenient method for their further characterization. These results show that treatment of cells with T interferon results in the induction of intracellular events identical to those observed in viral interferon-treated cells.     

Operant Conditioning of Heart Rate: Somatic Correlates

The relationships between heart rate (HR) and several parameters of somatic activity were evaluated in human subjects when shuck avoidance was made contingent on either increases or decreases in HR. In order to depict any influence of the contingency specific 10 HR, somatic activity was controlled to varying degrees by instructions and the use of non-contingent control groups. When increases in HR were reinforced, the contingency we found to influence somatic activity but an effect specific to HR was also observed. When decreases in HR were reinforced, there was no evidence that HR were influenced independently of somatic activity. The result are discussed with respect to several current issues.     

Stress,coping, family conflict,and adolescent alcohol use

This study examined alcohol use among seventh graders in relation to life events, daily hassles, the supportive quality of the family environment, coping, and anxiety. Four hundred twenty-five students participated, 228 girls and 197 boys. Stepwise regression and discriminant function analyses indicated that the students reported more alcohol use if they also reported more life events, more daily hassles, and more conflict in the family. A stress-buffering effect of low family conflict on life events could not be substantiated for extent of alcohol use. The results are discussed in the context of the developmental transitions of adolescence.     

Increased yield of full GBA sequencing in Ashkenazi Jews with Parkinson's disease

Background

Variants in GBA are the most common genetic risk factor for Parkinson's disease (PD), and are especially prevalent in the Ashkenazi Jewish (AJ) population. However, most studies on GBA in AJ genotype only seven selected Gaucher-associated pathogenic variants rather than sequencing the whole gene, which may leave carriers of PD-associated GBA variants undiscovered.

Change of tubular reabsorption of sodium and water after denervation

1. Renal function was compared in dogs before and after denervation, with normal or reduced glomerular filtration rate (GFR). GFR was reduced by one of two means, aortic clamping or injection of plastic microspheres into one renal artery.

2. The data showed that the tubular rejection fraction of denervated kidneys increased at a time when the same value for the control kidney decreased, indicating that denervation diuresis is not simply a result of increased filtered load of Na and water.

3. Changes in excretion after aortic clamping are not due solely to reduced filtered load. A reduction in renal blood pressure itself appears to have a direct effect on Na transport since Na excretion was significantly decreased before there were any changes in GFR, renal plasma flow and urine volume.

4. When GFR in a denervated kidney was reduced by microsphere injection it was demonstrated that a significant natriuresis (U_Na V) and diuresis (V) occurred when. GFR was reduced by as much as 40%.

5. These experiments support the hypothesis that denervation diuresis does not result solely from increased filtered load, but entails altered Na transport.