THE CLINICAL AND PARASITOLOGIC COURSES OF PLASMODIUM FALCIPARUM AND PLASMODIUM VIVAX INFECTIONS IN HUMANS: AN ANALYSIS OF 432 EPISODES OF INDUCED MALARIA IN THE JESSUP VOLUNTEER STUDIES, 1966-1975

The clinical records of 432 P. falciparum and P. vivax infected volunteer male inmates of the Maryland House of Corrections in Jessup, Maryland, were studied to determine (1) the clinical and parasitologic courses of infections in both parasite species, and (2) the influence of previous homologous and/or heterologous strain exposures on subsequent infections. The clinical and parasitologic courses of infection with both P. falciparum and P. vivax species indicated that: (a) there were characteristic strain related differences between P. falciparum and P. vivax. P. falciparum strains were more apt to cause severe infections than P. vivax strains. (b) Blood-induced infections produced significantly shorter prepatent and incubation periods than mosquito-induced. (c) Blacks tolerated the infections better than whites and, (d) homologous and heterologous strain immunities persisted with previous malaria history. In previously exposed cases, clinical manifestations were moderate, peak fever lowered, and peak parasitemias limited. (e) Anti-malarial drugs were effective in reducing sexual and asexual forms of the malaria parasite, and limiting peak fevers, irrespective of method of induction, race, parasite strain and species, and drug type used.^ Given these findings, and the current worldwide resurgence of malaria, this study has major implications in terms of setting malaria control and public health policies in both developed and developing countries.^

Effects of exercise with oxygen prebreathe for decompression risk reduction

Astronauts performing extravehicular activities (EVA) are at risk for occupational hazards due to a hypobaric environment, in particular Decompression Sickness (DCS). DCS results from nitrogen gas bubble formation in body tissues and venous blood. Denitrogenation achieved through lengthy staged decompression protocols has been the mainstay of prevention of DCS in space. Due to the greater number and duration of EVAs scheduled for construction and maintenance of the International Space Station, more efficient alternatives to accomplish missions without compromising astronaut safety are desirable. ^ This multi-center, multi-phase study (NASA-Prebreathe Reduction Protocol study, or PRP) was designed to identify a shorter denitrogenation protocol that can be implemented before an EVA, based on the combination of adynamia and exercise enhanced oxygen prebreathe. Human volunteers recruited at three sites (Texas, North Carolina and Canada) underwent three different combinations (“PRP phases”) of intense and light exercise prior to decompression in an altitude chamber. The outcome variables were detection of venous gas embolism (VGE) by precordial Doppler ultrasound, and clinical manifestations of DCS. Independent variables included age, gender, body mass index, oxygen consumption peak, peak heart rate, and PRP phase. Data analysis was performed both by pooling results from all study sites, and by examining each site separately. ^ Ten percent of the subjects developed DCS and 20% showed evidence of high grade VGE. No cases of DCS occurred in one particular PRP phase with use of the combination of dual-cycle ergometry (10 minutes at 75% of VO2 peak) plus 24 minutes of light EVA exercise (p = 0.04). No significant effects were found for the remaining independent variables on the occurrence of DCS. High grade VGE showed a strong correlation with subsequent development of DCS (sensitivity, 88.2%; specificity, 87.2%). In the presence of high grade VGE, the relative risk for DCS ranged from 7.52 to 35.0. ^ In summary, a good safety level can be achieved with exercise-enhanced oxygen denitrogenation that can be generalized to the astronaut population. Exercise is beneficial in preventing DCS if a specific schedule is followed, with an individualized VO2 prescription that provides a safety level that can then be applied to space operations. Furthermore, VGE Doppler detection is a useful clinical tool for prediction of altitude DCS. Because of the small number of high grade VGE episodes, the identification of a high probability DCS situation based on the presence of high grade VGE seems justified in astronauts. ^

FANCM AND FAAP24 MAINTAIN GENOMIC STABILITY THROUGH COOPERATIVE AND UNIQUE FUNCTIONS

Fanconi anemia (FA) is a rare recessive genetic disease with an array of clinical manifestations including multiple congenital abnormalities, progressive bone marrow failure and profound cancer susceptibility. A hallmark of cells derived from FA patients is hypersensitivity to DNA interstrand crosslinking agents such as mitomycin C (MMC) and cisplatin, suggesting that FA- and FA-associated proteins play important roles in protecting cells from DNA interstrand crosslink (ICL) damage. Two genes involved in the FA pathway, FANCM and FAAP24, are of particular interest because they contain DNA interacting domains. However, there are no definitive patient mutations for these two genes, and the resulting lack of human genetic model system renders their functional studies difficult. In this study, I established isogenic human FANCM- and FAAP24-null mutants through homologous replacement-mediated gene targeting in HCT-116 cells, and systematically investigated the functions of FANCM and FAAP24 inchromosome stability, FA pathway activation, DNA damage checkpoint signaling, and ICL repair. I found that the FANCM-/-/FAAP24-/- double mutant was much more sensitive to DNA crosslinking agents than FANCM-/- and FAAP24-/- single mutants, suggesting that FANCM and FAAP24 possess epistatic as well as unique functions in response to ICL damage. I demonstrated that FANCM and FAAP24 coordinately support the activation of FA pathway by promoting chromatin localization of FA core complex and FANCD2 monoubiqutination. They also cooperatively function to suppress sister chromatid exchange and radial chromosome formation, likely by limiting crossovers in recombination repair. In addition, I defined novel non-overlapping functions of FANCM and FAAP24 in response to ICL damage. FAAP24 plays a major role in activating ICL-induced ATR-dependent checkpoint, which is independent of its interaction with FANCM. On the other hand, FANCM promotes recombination-independent ICL repair independently of FAAP24. Mechanistically, FANCM facilitates recruitment of nucleotide excision repair machinery and lesion bypass factors to ICL damage sites through its translocase activity. Collectively, my studies provide mechanistic insights into how genome integrity is both coordinately and independently protected by FANCM and FAAP24.