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C Verhoef

Publications and source records attributed to C Verhoef.

31 records · Page 2Linked to original sources

Cloning and expression in Escherichia coli K-12 of the structural gene for outer membrane PhoE protein from Enterobacter cloacae.

In Escherichia coli K-12, the phoE gene encodes an outer membrane pore protein, which is induced by phosphate starvation. The corresponding gene of Enterobacter cloacae was transferred to E. coli K-12 by using RP4::mini Mu plasmid pULB113 and selecting for R-prime plasmids that carry the genes proA and proB, which are closely linked to phoE in E. coli K-12. The phoE gene was subcloned into the multicopy vector pACYC184, and the location of the gene was determined by analysis of in vitro constructed deletion plasmids and mutant plasmids generated by gamma delta insertions. The E. cloacae phoE gene is normally expressed in E. coli K-12, and the regulation of the expression is similar to that of the E. coli phoE gene. Functionally, the products of the phoE genes of E. coli K-12 and E. cloacae behave very similarly since they form pores in the outer membrane with a recognition site for negatively charged compounds and they serve as (part of) the receptor for phage TC45.

Bacterial Outer Membrane Proteins↗

Genetics and biochemistry of the peptidoglycan-associated proteins b and c of Escherichia coli K12.

To collect information on synthesis and regulation of the peptidoglycan-associated pore-forming outer membrane proteins b and c, mutants resistant to phages Me1 and TuIa were analyzed. Genetic analysis showed three linkage groups, corresponding with the genes tolF (phenotype b-c+), meoA (phenotype b+c-) and ompB (phenotypes b-c-, b-c+, b++c- and b++c+/-). It has recently been described that also a b+c- phenotype can occur in the latter linkage group [Chai, T., Foulds, J., J. Bacteriol. 130, 781-786 (1977)]. Among ompB (b-c+)/meoA (b+c-) double mutants strains were found with the b+c- phenotype, showing that ompB is not the structural gene for protein b. Studies on purified proteins b and c showed profound differences between the two proteins with respect to the electrophoretic mobility of fragments obtained by treatment with cyanogen bromide, trypsin and chymotrypsin. The amino acid in position three of the amino-termini of proteins b and c, isolated from isogenic strains, were identified as isoleucine and valine respectively. Both the genetic and biochemical results are consistent with a model recently published [Ichihara, S., Mizushima, S., J. Biochem. (Japan) 83, 1095-1100 (1978)] which predicts that tolF and meoA are the structural genes for the proteins b and c respectively and that ompB is a regulatory gene whose product regulates the levels of both proteins.

Bacterial Proteins↗

meoA is the structural gene for outer membrane protein c of Escherichia coli K12.

The isolation and characterization of two mutants of Escherichia coli K12 with an altered outer membrane protein c is described. The first mutant, strain CE1151, was isolated as a bacteriophage Me1 resistant strain which contains normal levels of protein c. Mutant cells adsorbed the phage with a strongly decreased rate. Complexes of purified nonheat modified wild type protein c and wild type lipopolysaccharide inactivated phage Me1, indicating that these components are required for receptor activity for phage Me1. When wild type protein c was replaced by protein c of strain CE1151, the receptor-complex was far less active, showing that protein c of strain CE1151 is altered. The second mutant produces a protein c with a decreased electrophoretic mobility, designated as protein c. An altered apparent molecular weight was also observed for one or more fragments obtained after fragmentation of the mutant protein with cyanogen bromide, trypsin and chymotrypsin. Alteration of protein c was not accompanied by a detectable alteration in protein b or its fragments. Both mutations are located at minute 48 of the Escherichia coli K12 linkage map. The results strongly suggest that meoA is the structural gene for protein c.

Bacterial Proteins↗

Mapping of a gene for a major outer membrane protein of Escherichia coli K12 with the aid of a newly isolated bacteriophage.

A method is described for the enrichment of phages which can adsorb to a specific determinant of bacterial cell surfaces. A phage was isolated which absorbs to E. coli cells containing the "major outer membrane= protein c but not to strains that are lacking this protein. With the aid of this phage a gene, meoA which is responsible for the lack of protein c was mapped at 48 min on the linkage map of E. coli K12.

Bacterial Proteins↗

Radiofrequency ablation in patients with primary and secondary hepatic malignancies.

The aims of this study were to assess the technical effectiveness of radiofrequency (RF) ablation in patients with primary or secondary hepatic malignancies and to determine survival and complication rates. This was a retrospective analysis of prospectively collected data of patients treated with RF ablation and controlled for recurrence every 3 months by contrast-enhanced computed tomography (CT) or magnetic resonance imaging (MRI). The outcome is compared with a comprehensive review of data published in recent literature. Forty-seven patients underwent 50 RF sessions for the ablation of 73 tumors. Local tumor progression was observed in 11 patients (23%). A tumor sized larger than 30 mm, a tumor load larger than 14 cm3, and a percutaneous approach were associated with a faster time to local tumor progression. At the end of a mean (+/- SD) follow-up period of 11.4 +/- 7.5 months, 39 patients (83%) were alive, including eight patients with recurrent disease. The overall cumulative survival rates at 12 and 24 months were 87% and 70%, respectively. In our center, RF ablation can be safely performed to achieve adequate local control and survival rates. Time to local tumor progression was significantly related to initial size of the tumor and tumor load.

Adult↗

Resection of extrahepatic hepatocellular carcinoma metastasis can result in long-term survival.

Hepatocellular carcinoma (HCC) is one of the most common primary cancers in the world and the third most common cause of cancer mortality world-wide. Surgery is the gold standard in the treatment of patients with HCC. The prognosis is mainly determined by the underlying liver disease and recurrent rates. In the Western World, up to 30% of the patients with HCC have a non-cirrhotic liver. The main prognostic factor in this special group of patients are the recurrences. Most recurrences are intrahepatic; however, 30% of the recurrences are extrahepatic. The role of resection in case of intrahepatic recurrences is widely accepted, particularly in the non-cirrhotic liver. The role of resection in extrahepatic HCC recurrences is not well established and unknown among many physicians. We present two patients with HCC in a non-cirrhotic liver with extrahepatic recurrences and long-term survival after resection. The corresponding literature support an aggressive approach in case of extrahepatic HCC recurrence in selected cases: resectable metastasis, preserved liver function, absence of intracranial metastasis and control of the primary tumour. Further research is warranted because of the limited number of reports and the absence of randomized trials.

Adult↗