Downeast anemia (dea), a new mouse model of severe nonspherocytic hemolytic anemia caused by hexokinase (HK(1)) deficiency.

A new spontaneous mutation in the A/J inbred mouse strain, downeast anemia (dea), causes severe hemolytic anemia with extensive tissue iron deposition and marked reticulocytosis. The anemia is present at birth and persists throughout life. The defect is inherited as an autosomal recessive and is transferable through bone marrow stem cells. The red cell morphology is consistent with a nonspherocytic hemolytic anemia, suggestive of a red cell enzymopathy. In linkage analysis, dea is nonrecombinant with the hexokinase-1 gene (Hk1) on mouse Chromosome 10. Expression of Hk1 is markedly decreased in dea erythroid tissues, and the transcript produced is larger than normal. Hexokinase enzyme activity is significantly decreased in dea tissues, including red cells, spleen, and kidney. Southern blot analyses revealed approximately 5.5 kb of additional sequence in the 5' portion of the dea Hk1 gene, which was identified by direct sequencing as an early transposon (ETn) insertion in intron 4. ETn insertions disrupt genes in several mouse models by a variety of mechanisms, including aberrant splicing of ETn sequences into the mRNA. We conclude that the primary gene defect in the dea mutation is in Hk1 and that dea is a model of generalized hexokinase deficiency, the first such model identified to date.

Defects in the cappuccino (cno) gene on mouse chromosome 5 and human 4p cause Hermansky-Pudlak syndrome by an AP-3-independent mechanism.

Defects in a triad of organelles (melanosomes, platelet granules, and lysosomes) result in albinism, prolonged bleeding, and lysosome abnormalities in Hermansky-Pudlak syndrome (HPS). Defects in HPS1, a protein of unknown function, and in components of the AP-3 complex cause some, but not all, cases of HPS in humans. There have been 15 inherited models of HPS described in the mouse, underscoring its marked genetic heterogeneity. Here we characterize a new spontaneous mutation in the mouse, cappuccino (cno), that maps to mouse chromosome 5 in a region conserved with human 4p15-p16. Melanosomes of cno/cno mice are immature and dramatically decreased in number in the eye and skin, resulting in severe oculocutaneous albinism. Platelet dense body contents (adenosine triphosphate, serotonin) are markedly deficient, leading to defective aggregation and prolonged bleeding. Lysosomal enzyme concentrations are significantly elevated in the kidney and liver. Genetic, immunofluorescence microscopy, and lysosomal protein trafficking studies indicate that the AP-3 complex is intact in cno/cno mice. It was concluded that the cappuccino gene encodes a product involved in an AP-3-independent mechanism critical to the biogenesis of lysosome-related organelles. (Blood. 2000;96:4227-4235)

Emergence of Streptococcus faecalis isolates with high-level resistance to multiple aminocyclitol aminoglycosides.

Isolates of Streptococcus faecalis were occasionally found to be resistant to gentamicin concentrations of 500 micrograms/ml in a microdilution tray. Additional tests were performed with 28 such isolates. They all demonstrated high-level resistance (minimum inhibitory concentration greater than 2000 micrograms/ml) to gentamicin, kanamycin, sisomicin, streptomycin, and tobramycin. This high-level aminoglycoside resistance was associated with total resistance to in vitro synergism when each aminoglycoside was combined with ampicillin. In addition to ampicillin-aminoglycoside combinations, ampicillin-vancomycin, ampicillin-rifampin, rifampin-vancomycin, and vancomycin-gentamicin combinations were also tested; all failed to exhibit an in vitro synergistic-bactericidal effect against these enterococci. The emergence of enterococci with high-level resistance to multiple aminoglycosides in this clinical population is a source of grave concern with obvious therapeutic implications in situations such as endocarditis, where an in vitro bactericidal effect is thought to be necessary for a cure. No drug or combination of drugs was found to be bactericidal for these isolates. It is suspected that similar isolates of group D streptococci with high-level resistance to gentamicin may emerge in other institutions but go unrecognized. Surveillance by others for identifying this newly emerging pathogen carrying multiple resistance and a continued search for a satisfactory chemotherapeutic agent(s) are encouraged.