WHOLE GENOME TARGETED ENRICHMENT AND SEQUENCING OF HUMAN-INFECTING CRYPTOSPORIDIUM spp.

Cryptosporidium spp. are protozoan parasites that cause severe illness in vulnerable human populations. Obtaining pure Cryptosporidium DNA from clinical and environmental samples is challenging because the oocysts shed in contaminated feces are limited in quantity, difficult to purify efficiently, may derive from multiple species, and yield limited DNA (<40 fg/oocyst). Here, we develop and validate a set of 100,000 RNA baits (CryptoCap_100k) based on six human-infecting Cryptosporidium spp. ( C. cuniculus , C. hominis , C. meleagridis , C. parvum , C. tyzzeri , and C. viatorum ) to enrich Cryptosporidium spp. DNA from a wide array of samples. We demonstrate that CryptoCap_100k increases the percentage of reads mapping to target Cryptosporidium references in a wide variety of scenarios, increasing the depth and breadth of genome coverage, facilitating increased accuracy of detecting and analyzing species within a given sample, while simultaneously decreasing costs, thereby opening new opportunities to understand the complex biology of these important pathogens.

A measurement of the local energy deposition by antiprotons coming to rest in tissue-like material.

The measurement of the energy deposited in tissue-equivalent plastic as a beam of low-energy antiprotons slows down is described. The resulting depth-dose curve is presented and compared with that of protons measured under similar conditions. These curves indicate that the ratio of the dose in the antiproton stopping peak to that in the plateau region is about twice that found for protons. Analysis of the data shows that out of the 2 GeV available in an antiproton annihilation, on average less than 30 MeV is deposited close to the site of the interaction. This leads to the conclusion that antiprotons are unlikely to offer any marked advantage over other radiations for radiotherapeutic applications of the future.

The biological effect of very high energy hadrons.

The meristems of Vicia faba bean roots were irradiated at different depths in water in a 250 GeV/c positive hadron beam from the CERN Super Proton Synchrotron (SPS). The growth during the 10 days following irradiation was determined and compared with results obtained from 60Co gamma-ray exposures. The data indicate a Relative Biological Efficiency (r.b.e.) near to one, independent of the contribution of secondaries to the dose. However, some anomalies in the radiation response of the irradiated beans were noted.

RBE and OER values of negative pion beams from growth inhibition of Vicia faba roots.

Two pion beams of different momentum width have been used to expose meristems of Vicia Faba roots under aerobic and hypoxic conditions. The measurements of the resulting 10 days growth inhibition after exposures at various locations on the pion beam axes have been made and RBE and OER values evaluated for 50% effects compared to 60Co gamma-rays. The results have been related to the fractional doses from star products defined by telescope measurements of stopped pions along the same beams. It has been found that the RBE value increases with the fractional "star dose" up to a maximum after which the RBE decreases. The OER values, however, were found to decrease with increasing "star dose" fraction rather rapidly after which it was found to be independent of the "star dose" contribution.

Effects of negative pions and neutrons on the growth of Vicia faba bean roots.

Vicia faba bean roots have been irratiated with neutrons of various energies and with negative pi-mesons, and the effect on the ten-day growth of the roots has been determened. The neutron irratiations were made in beams of 400 and 600 MeV maximum energy, as well as with neutrons from a plutonium-beryllium source (mean energy 4.4 MeV) and from a 14 MeV neutron generator. The bean roots have also been irradiated at various points along the depth-dose curve of negative pi-mesons, including the gegion where the pions annihilate on coming to rest. The results show a maximum relative biological effectiveness (RBE) of 3.7 for 50% reduction in ten days growth for stopped negative pions and values up to 3.3 for high-energy neutrons, compared to 5.5 for 14 MeV neutrons. The biological effectiveness of high-energy neutrons and stopped pions shows a more pronounced dependence on dose than does the effect with lower-energy neutrons.

Determination of some parameters for pion radiobiology studies.

An experimental investigation of the central axis depth-dose and stopping rate distribution of the SIN biomedical pion beam is reported. The pion stopping rate in a thin disc of tissue-equivalent plastic was determined using a counter telescope. The dose rate at the position of this dic 'target' was measured using a specially designed parallel-plate tissue-equivalent ionization chamber. Both dose rate and pion stopping rate are given as a function of depth for beams of two different momenta spread. The energy deposition required per pion stop to fit the measured dose rate curves was calculated and found to be between 37 and 40 MeV. From the stopping rate measurements the depth-dose distribution of pion interaction dose (star-dose) and the dose due to the pion slowing down have been evaluated. The maximum star-dose is found at a depth of about 2 g cm-2 beyound the maximum of the ionization dose.