Transformed phenotype conferred to NIH/3T3 cells by ectopic expression of heparin-binding growth factor 1/acidic fibroblast growth factor

Summary Heparin-binding growth factor 1 (HBGF-1), also known as acidic fibroblast growth factor, is a potent mitogen and angiogenic factor found in tissues such as brain, kidney and heart. The genomic and cDNA sequences indicate that HBGF-1 does not have a typical signal peptide sequence. HBGF-1 was shown to be localized to the extracellular matrix of cardiac myocytes, but the mechanism of secretion is not presently known. We have cloned the HBGF-1 cDNA which allowed us to directly test the biological activity, mechanism of secretion and transforming potential of the recombinant protein. A previous report showed that the truncated HBGF-1 confers partial transformed phenotype to the recipient fibroblasts. However, expression of full-length HBGF-1 has not been reported. The HBGF-1 coding sequence was cloned into the retroviral expression vector, SVX, and transfected into NIH/3T3 cells. Transfectants expressing full-length HBGF-1 protein at high levels form foci and grow to a higher cell density than the parental NIH/3T3 cells. Western blotting analysis showed that the recombinant HBGF-1 is a unique band of approximately 20 kDa and can be detected in the cell homogenate but not in the conditioned medium. NIH/3T3 cells were conferred anchorage independence when HBGF-1 was provided exogenously. We showed the transformed cells are capable of growing on soft agar even in the absence of exogenously-provided HBGF-1. Transfected cells expressing HBGF-1 also induced tumor formation when injected into nude mice. Thus, NIH/3T3 cells acquired a full spectrum of transformed phenotype when full length HBGF-1 was expressed at high levels. This work was supported by grants from the National Cancer Institute (RO1 CA45611), The March of Dimes Birth Defects Foundation (No. 6-549) and The Ohio Cancer Research Associates, Inc. I.-M.C. is a recipient of The Research Career Development Award (KO4 CA01369) from the National Institutes of Health.     

A medium for the detection of Lancefield Group D cocci in skimmed milk powder by measurement of conductivity changes

A selective medium for the detection of Lancefield Group D cocci in skimmed milk powder by conductivity measurements was developed and evaluated using the Bactometer M123 and Malthus 128H systems. This medium promoted large changes in conductance and capacitance. The calibration curve of detection times vs concentration of Lancefield Group D cocci showed a linear correlation coefficient of 0.93 and the method gave comparable results in both conductivity instruments. Naturally contaminated samples containing c. 10(3) cfu/g of Lancefield Group D cocci gave detection times within 16-18 h which was sufficiently rapid for the medium to be used for the routine screening of skimmed milk powder.     

The identification of T2; the phosphate/pyrophosphate transport protein of the hepatic microsomal glucose-6-phosphatase system

The phosphate/pyrophosphate translocase protein (T2) of the hepatic microsomal glucose-6-phosphatase system was identified and then purified using antibodies raised against the rat mitochondrial phosphate/hydroxyl ion antiport protein. The T2 protein was shown to be absent in the microsomes isolated from a patient previously diagnosed as having type lc glycogen storage disease.     

Waveform discrimination in a pair of pulse-generating electric fishes

Studies of pulse-type gymnotiform electric fishes have suggested that electric organ discharge waveforms (EODw) allow individuals to discriminate between conspecific and allospecific signals, but few have approached this experimentally. Here we implement a phase-locked playback technique for a syntopic species pair, Brachyhypopomus gauderio and Gymnotus omarorum. Both species respond to changes in stimulus waveform with a transitory reduction in the interpulse interval of their self-generated discharge, providing strong evidence of discrimination. We also document sustained rate changes in response to different EODws, which may suggest recognition of natural waveforms.     

Symptoms and signs: physical disease or illness behaviour?

The amount of treatment received by 380 patients with backache was found to have been influenced more by their distress and illness behaviour than by the actual physical disease. Patients showing a large amount of inappropriate illness behaviour had received significantly more treatment (p less than 0.001). The symptoms and signs of illness behaviour need to be clearly distinguished from those of physical disease, and better assessment of illness behaviour is essential if everyday clinical practice is to fulfil the ideal of treating patients as well as diseases.     

Distribution and effect of the weak acids 5,5-dimethyl-2,4-oxazolidinedione (DMO) and 2,4-dinitrophenol (DNP) in membrane vesicles of Micrococcus denitrificans.

The effects of 5,5-dimethyl-2,4-oxazolidinedione (DMO) and 2,4-dinitrophenol (DNP) on membrane vesicles of Micrococcus denitrificans were compared. DMO did not affect the ability of these vesicles to accumulate glycine in the presence of the substrate l-lactate. Both glycine transport and l-lactate oxidation were inhibited by DNP; the concentration of DNP required for inhibition of respiration was fortyfold higher than that required for inhibition of transport. Using the technique of equilibrium dialysis with membrane residues from which the lipid had been extracted, no binding of [¹⁴C]DMO to membrane protein was detected. However, [¹⁴C]DNP did bind to membrane protein. At 100 μm DNP, 12% of the [¹⁴C]DNP was bound, equivalent to 1.56 nmol/mg protein. The pH inside vesicles respiring on l-lactate was calculated from the distribution of [¹⁴C]DMO and was found not to differ from the pH of the suspending buffer. The mechanism of action of DNP on active transport in M. denitrificans vesicles appears not to involve proton conduction.