Tigecycline resistance has been attributed to ramA overexpression and subsequent acrA upregulation. The ramA locus, originally identified in Klebsiella pneumoniae, has homologues in Enterobacter and Salmonella spp. In this study, we identify in silico that the ramR binding site is also present in Citrobacter spp. and that Enterobacter, Citrobacter and Klebsiella spp. share key regulatory elements in the control of the romA–ramA locus. RACE (rapid amplification of cDNA ends) mapping indicated that there are two promoters from which romA–ramA expression can be regulated in K. pneumoniae. Correspondingly, electrophoretic binding studies clearly showed that purified RamA and RamR proteins bind to both of these promoters. Hence, there appear to be two RamR binding sites within the Klebsiella romA–ramA locus. Like MarA, RamA binds the promoter region, implying that it might be subject to autoregulation. We have identified changes within ramR in geographically distinct clinical isolates of K. pneumoniae. Intriguingly, levels of romA and ramA expression were not uniformly affected by changes within the ramR gene, thereby supporting the dual promoter finding. Furthermore, a subset of strains sustained no changes within the ramR gene but which still overexpressed the romA–ramA genes, strongly suggesting that a secondary regulator may control ramA expression.

In the last decade tuberculosis (TB) has reemerged as a major worldwide public health hazard with increasing incidence among adults and children. Although cases among children represent a small percentage of all TB cases, infected children are a reservoir from which many adult cases will arise. TB diagnosis in children usually follows discovery of a case in an adult, and relies on tuberculin skin testing, chest radiograph, and clinical signs and symptoms. However, clinical symptoms are nonspecific, skin testing and chest radiographs can be difficult to interpret, and routine laboratory tests are not helpful. Although more rapid and sensitive laboratory testing, which takes into account recent advances in molecular biology, immunology, and chromatography, is being developed, the results for children have been disappointing. Better techniques would especially benefit children and infants in whom early diagnosis is imperative for preventing progressive TB.

Macroprolactinomas commonly cause pressure effects on immediate parasellar structures, in particular on the optic chiasm to cause visual field defects. Pressure on more distant brain structures is rarely reported. We describe two massive prolactinomas presenting with neurological signs, including signs of hemiparesis which, to our knowledge, has not been reported previously.

Images

We present a case of adult hepatoblastoma. This young female presented with severe acute cholangitis. Preoperative diagnosis was common bile duct (CBD) obstruction with portal vein thrombosis. On exploration she had a tumor mass in the CBD. The unusual features of this case are discussed in this report.

Integration of retroviral DNA into the host chromosome requires a virus-encoded integrase (IN). IN recognizes, cuts and then joins specific viral DNA sequences (LTR ends) to essentially random sites in host DNA. We have used computer-assisted protein alignments and mutagenesis in an attempt to localize these functions within the avian retroviral IN protein. A comparison of the deduced amino acid sequences for 80 retroviral/retrotransposon IN proteins reveals strong conservation of an HHCC N-terminal 'Zn finger'-like domain, and a central D(35)E region which exhibits striking similarities with sequences deduced for bacterial IS elements. We demonstrate that the HHCC region is not required for DNA binding, but contributes to specific recognition of viral LTRs in the cutting and joining reactions. Deletions which extend into the D(35)E region destroy the ability of IN to bind DNA. Thus, we propose that the D(35)E region may specify a DNA-binding/cutting domain that is conserved throughout evolution in enzymes with similar functions.

Images