Molecular characterization of PulE, a protein required for pullulanase secretion

pulE, one of 14 genes specifically required for pullulanase secretion in Klebsiella oxytoca, codes for a putative nucleotide-binding protein. Subcellular fractionation indicated that the majority of PulE in Escherichia coli cells expressing all 14 secretion genes is mainly associated with the cytoplasmic membrane through both hydrophobic and non-hydrophobic interactions. Mutational analysis revealed that one of the two regions of PulE that are conserved in many nucleotide-binding proteins (Walker box A) is essential for pullulanase secretion. Likewise, mutations that removed aspartate residues from each of two regions immediately downstream from the Walker box A also reduced secretion. These aspartate-rich regions are highly conserved in all 16 known PulE homologues but not in any other nucleotide-binding proteins. Altogether, these results indicate that PulE might belong to a new family of nucleotide-binding proteins. The protein could not be cross-linked to the photoactivatable ATP analogue azido-ATP, however. Most pulE point or deletion mutations which prevented pullulanase secretion exhibited transdominance when expressed at high levels in cells producing wild-type PulE protein. Evidence presented suggests that PulE might be a homodimer.     

Postoperative electromyographic monitoring of small intestine function in patients with peritonitis and acute intestinal obstruction

In 49 patients with general peritonitis and acute ileus, the authors used the method of direct electromyography. On the basis of analysis of the results, the functional state of the intestine, dynamics of its motor activity, effectiveness of the treatment performed were assessed and outcome of the disease was predicted.     

Instability in the ATCT variable number tandem repeat locus VWF.VNTR I in intron 40 of von Willebrand factor gene

The von Willebrand factor gene intron 40 variable number tandem repeat VWF.VNTR I exhibits 10 alleles making it highly polymorphic and useful for parentage and forensic testing, 45 unrelated families (210 meiotic events) were tested for VWF.VNTR I alleles. One spontaneous mutation was observed in a family member. Haplotype analysis demonstrated that this mutation was due to a gain of one motif repeat by a paternal allele. Sequence analysis confirmed the difference in the number of motif repeats between the proband and the alleles expressed by the parents. This instability emphasizes the importance of demonstrating exclusion in at least two separate loci in parentage testing.     

The therapeutic efficacy of interferon-alfa-2a (Roferon-A) in chronic hepatitis C

THE AIM OF THE STUDY: Evaluation of 6-month treatment with roferon-A of patients with chronic hepatitis C (CHC) and comparison of the treatment regimens. MATERIAL AND METHODS: 79 CHC patients received roferon-A for 3 months in a dose 6,000,000 IU 3 time a week. In case of the response to treatment was continued for the next 3 months (3,000,000 IU 3 times a week). Clinical-laboratory findings, results, of EIA and liver biopsy histology were examined. RESULTS: Primary remission was achieved in 71.8% of cases. 36.9% of patients developed recurrences including 14.1% recurrence arising in the course of therapy. Stable remission was obtained in 32.4% of patients. 28.2% patients were non-responders. Side effects were mild, discontinuation of the treatment was necessary only in 7% of cases. CONCLUSION: Roferon-A administration in a dose 6,000,000 IU for 24 weeks is optimal both as related to cost and efficacy of the treatment.