BIOLOGICAL AVAILABILITY OF IRON TO THE FRESHWATER CYANOBACTERIUM ANABAENA FLOS‐AQUAE1

Iron acquisition from various ferric chelates and colloids was studied using iron‐limited cells of Anabaena flos‐aquae (Lyng.) Brèb UTEX 1444, a cyanobacterial strain that produces high levels of siderophores under iron limitation. Various chelators of greatly varying affinity for Fe³⁺ (HEDTA, EDDHA, desferrioxamine mesylate, HBED, 8‐hydroxyquinoline) were assayed for the degree of iron acquisition by iron‐limited cyanobacterial cells. Iron uptake rates (measured by graphite furnace atomic absorption spectrometry) varied approximately inversely with calculated [Fe³⁺] (calculated as pFe) and decreased with increasing chelator‐to‐iron ratio. No iron uptake was observed when Fe³⁺ was chelated with HBED, the strongest of the tested chelators. Iron‐limited Anabaena cells were able to take up iron from 8‐hydroxyquinoline (oxine or 8HQ), a compound sometimes used to quantify aquatic iron bioavailability. Iron bound to purified humic acid was poorly available but did support some growth at high humic acid concentrations. These results suggest that for cyanobacteria, even tightly bound iron is biologically available, including to a limited extent iron bound to humic acids. However, iron bound to some extremely strong chelators (e.g. HBED) is likely to be biologically unavailable.     

FUNGUS DISEASE IN NORTHERN CALIFORNIA

A systematic search for disease caused by fungi was made in a group of patients from northern California.Superficial mycotic infection was diagnosed frequently. Systemic fungus infection was found in approximately 0.6 per cent of an unselected group observed at Stanford University clinics—coccidioidomycosis or actinomycosis in almost all cases.Clinically unsuspected fungus infection was rare. Various species of Candida were frequently recovered, particularly from debilitated patients who had received antimicrobial therapy. Only rarely could these organisms be conclusively demonstrated to have caused serious disease.     

Genome-wide association and linkage analyses localize a progressive retinal atrophy locus in Persian cats

Hereditary eye diseases of animals serve as excellent models of human ocular disorders and assist in the development of gene and drug therapies for inherited forms of blindness. Several primary hereditary eye conditions affecting various ocular tissues and having different rates of progression have been documented in domestic cats. Gene therapy for canine retinopathies has been successful, thus the cat could be a gene therapy candidate for other forms of retinal degenerations. The current study investigates a hereditary, autosomal recessive, retinal degeneration specific to Persian cats. A multi-generational pedigree segregating for this progressive retinal atrophy was genotyped using a 63 K SNP array and analyzed via genome-wide linkage and association methods. A multi-point parametric linkage analysis localized the blindness phenotype to a ~1.75 Mb region with significant LOD scores (Z ≈ 14, θ = 0.00) on cat chromosome E1. Genome-wide TDT, sib-TDT, and case–control analyses also consistently supported significant association within the same region on chromosome E1, which is homologous to human chromosome 17. Using haplotype analysis, a ~1.3 Mb region was identified as highly associated for progressive retinal atrophy in Persian cats. Several candidate genes within the region are reasonable candidates as a potential causative gene and should be considered for molecular analyses.     

Cyclophilins Facilitate Dissociation of the Human Papillomavirus Type 16 Capsid Protein L1 from the L2/DNA Complex following Virus Entry

Human papillomaviruses (HPV) are composed of the major and minor capsid proteins, L1 and L2, that encapsidate a chromatinized, circular double-stranded DNA genome. At the outset of infection, the interaction of HPV type 16 (HPV16) (pseudo)virions with heparan sulfate proteoglycans triggers a conformational change in L2 that is facilitated by the host cell chaperone cyclophilin B (CyPB). This conformational change results in exposure of the L2 N terminus, which is required for infectious internalization. Following internalization, L2 facilitates egress of the viral genome from acidified endosomes, and the L2/DNA complex accumulates at PML nuclear bodies. We recently described a mutant virus that bypasses the requirement for cell surface CyPB but remains sensitive to cyclosporine for infection, indicating an additional role for CyP following endocytic uptake of virions. We now report that the L1 protein dissociates from the L2/DNA complex following infectious internalization. Inhibition and small interfering RNA (siRNA)-mediated knockdown of CyPs blocked dissociation of L1 from the L2/DNA complex. In vitro, purified CyPs facilitated the dissociation of L1 pentamers from recombinant HPV11 L1/L2 complexes in a pH-dependent manner. Furthermore, CyPs released L1 capsomeres from partially disassembled HPV16 pseudovirions at slightly acidic pH. Taken together, these data suggest that CyPs mediate the dissociation of HPV L1 and L2 capsid proteins following acidification of endocytic vesicles.     

Pathogen factors and host factors in murine pulmonary blastomycosis

Factors related to pathogenesis were studied in a murine model of pulmonary fungal infection with strains of Blastomyces dermatitidis. Strain 26199V was 10⁵-fold more lethal in pulmonary challenge than strain GA-1. This relationship between the two strains also occurred after intraperitoneal challenge. Serial studies with lung cultures and histology of sacrificed animals after pulmonary challenge with similar doses, and challenge studies in mice of differing maturity, indicated GA-1 is impaired in initial replication in situ, although the two strains grow equally well in the presence of murine substrates in vitro. In vitro studies indicated the two strains were indistinguishable in most studies of biochemical and metabolic markers, and in studies with chemical and physical inhibitors. The exceptions were their morphologic appearance in the yeast phase, ability to hydrolyze esculin, and susceptibility to crystal violet dye and bile. The last finding and previous work suggested possible differences in lipid content; these were confirmed in studies which also included two other strains. Greater phospholipid and palmitic acid content were associated with greater virulence.     

Passive immunization in murine mucormycosis

Antibody raised in mice against mycelial homogenates of Rhizomucor pusillus was effective in passive immunization against pulmonary and disseminated mucormycosis (phycomycosis) in immunocompromised mice. Mice intranasally inoculated and infected with Rh. pusillus and treated with antisera had a statistically significant increased resistance to infection and a diminished secondary dissemination of viable fungal fragments. Histological examination of infected lung tissues showed that antibody treated animals were apparently able to degrade hyphal fragments.     

Cryptococcus Neoformans in Pigeon Feces in San Francisco

Typical Cryptococcus neoformans was isolated from one of 10 specimens of pigeon feces collected in downtown San Francisco. This isolation from a small sample suggests considerable prevalence of this important pathogen and tends to confirm that it is ubiquitous.