18F-EF5 PET Is Predictive of Response to Fractionated Radiotherapy in Preclinical Tumor Models

We evaluated the relationship between pre-treatment positron emission tomography (PET) using the hypoxic tracer ¹⁸F-[2-(2-nitro-1-H-imidazol-1-yl)-N-(2,2,3,3,3- pentafluoropropyl) acetamide] (¹⁸F-EF5) and the response of preclinical tumor models to a range of fractionated radiotherapies. Subcutaneous HT29, A549 and RKO tumors grown in nude mice were imaged using ¹⁸F-EF5 positron emission tomography (PET) in order to characterize the extent and heterogeneity of hypoxia in these systems. Based on these results, 80 A549 tumors were subsequently grown and imaged using ¹⁸F-EF5 PET, and then treated with one, two, or four fraction radiation treatments to a total dose of 10–40 Gy. Response was monitored by serial caliper measurements of tumor volume. Longitudinal post-treatment ¹⁸F-EF5 PET imaging was performed on a subset of tumors. Terminal histologic analysis was performed to validate ¹⁸F-EF5 PET measures of hypoxia. EF5-positive tumors responded more poorly to low dose single fraction irradiation relative to EF5-negative tumors, however both groups responded similarly to larger single fraction doses. Irradiated tumors exhibited reduced ¹⁸F-EF5 uptake one month after treatment compared to control tumors. These findings indicate that pre- treatment ¹⁸F-EF5 PET can predict the response of tumors to single fraction radiation treatment. However, increasing the number of fractions delivered abrogates the difference in response between tumors with high and low EF5 uptake pre-treatment, in agreement with traditional radiobiology.     

Dynamic remodeling of the actin cytoskeleton: Lessons learned from Listeria locomotion

The bacterial pathogen Listeria monocytogenes displays the remarkable ability to reorganize the actin cytoskeleton within host cells as a means for promoting cell-to-cell transfer of the pathogen, in a manner that evades humoral immunity. In a series of events commencing with the biosynthesis of the bacterial surface protein ActA, host cell actin and many actin-associated protein self-assemble to from rocket-tail structures that continually grow at sites proximal to the bacterium and depolymerize distally. Widespread interest in the underlying molecular mechanism of Listeria locomotion stems from the likelihood that the dynamic remodeling of the host cell actin cytoskeleton at the cell's leading edge involves mechanistically analogous interactions. Recent advances in our understanding of these fundamental cytoskeletal rearrangements have been achieved through a clearer recognition of the central role of oligo-proline sequence repeats present in ActA, and these findings provide a basis for inferring the role of analogous host cell proteins in the force-producing and position-securing steps in pseudopod and lamellipod formation at the peripheral membrane.     

Nutrients Limiting Soybean (glycine max l) Growth in Acrisols and Ferralsols of Western Kenya

Low soybean yields in western Kenya have been attributed to low soil fertility despite much work done on nitrogen (N) and phosphorus (P) nutrition leading to suspicion of other nutrient limitations. To investigate this, a nutrient omission trial was set up in the greenhouse at the University of Eldoret-Kenya to diagnose the nutrients limiting soybean production in Acrisols from Masaba central and Butere sub-Counties, and Ferralsols from Kakamega (Shikhulu and Khwisero sub-locations) and Butula sub-Counties and to assess the effect of liming on soil pH and soybean growth. The experiment was laid out in a completely randomized design with ten treatments viz; positive control (complete), negative control (distilled water), complete with lime, complete with N, minus macronutrients P, potassium (K), calcium (Ca), magnesium (Mg) and sulphur (S) and with, micro-nutrients boron (B), molybdenum (Mo), manganese (Mn), copper (Cu) and zinc (Zn) omitted. Visual deficiency symptoms observed included interveinal leaf yellowing in Mg omission and N addition and dark green leaves in P omission. Nutrients omission resulted in their significantly low concentration in plant tissues than the complete treatment. Significantly (P≤ 0.05) lower shoot dry weights (SDWs) than the complete treatment were obtained in different treatments; omission of K and Mg in Masaba and Shikhulu, Mg in Khwisero, K in Butere and, P, Mg and K in Butula. Nitrogen significantly improved SDWs in soils from Kakamega and Butula. Liming significantly raised soil pH by 9, 13 and 11% from 4.65, 4.91 and 4.99 in soils from Masaba, Butere and Butula respectively and soybean SDWs in soils from Butere. The results show that, poor soybean growth was due to K, Mg and P limitation and low pH in some soils. The results also signify necessity of application of small quantities of N for initial soybean use.     

Lymphocytic Choriomeningitis Virus Infection in FVB Mouse Produces Hemorrhagic Disease

The viral family Arenaviridae includes a number of viruses that can cause hemorrhagic fever in humans. Arenavirus infection often involves multiple organs and can lead to capillary instability, impaired hemostasis, and death. Preclinical testing for development of antiviral or therapeutics is in part hampered due to a lack of an immunologically well-defined rodent model that exhibits similar acute hemorrhagic illness or sequelae compared to the human disease. We have identified the FVB mouse strain, which succumbs to a hemorrhagic fever-like illness when infected with lymphocytic choriomeningitis virus (LCMV). FVB mice infected with LCMV demonstrate high mortality associated with thrombocytopenia, hepatocellular and splenic necrosis, and cutaneous hemorrhage. Investigation of inflammatory mediators revealed increased IFN-γ, IL-6 and IL-17, along with increased chemokine production, at early times after LCMV infection, which suggests that a viral-induced host immune response is the cause of the pathology. Depletion of T cells at time of infection prevented mortality in all treated animals. Antisense-targeted reduction of IL-17 cytokine responsiveness provided significant protection from hemorrhagic pathology. F1 mice derived from FVB×C57BL/6 mating exhibit disease signs and mortality concomitant with the FVB challenged mice, extending this model to more widely available immunological tools. This report offers a novel animal model for arenavirus research and pre-clinical therapeutic testing.     

CLAN, a Novel Human CED-4-like Gene

Proteins governing cell death form the basis of many normal processes and contribute to the pathogenesis of many diseases when dysregulated. Here we report the cloning of a novel human CED-4-like gene, CLAN, and several of its alternatively spliced isoforms. These caspase-associated recruitment domain (CARD)-containing proteins are expressed at varying degrees in normal human tissues and may contribute to a number of intracellular processes including apoptosis, cytokine processing, and NF-κB activation. The CARD of the CLAN proteins binds a number of other CARD-containing proteins including caspase-1, BCL10, NOD2, and NAC. Once their physiologic functions are uncovered, CLAN proteins may prove to be valuable therapeutic targets.     

Skin banking methodology: An evaluation of package format,cooling and warming rates,and storage efficiency

The two major skin packaging formats for transplantable human skin, flat — folded and rolled — cylindrical, were evaluated with respect to the control of cooling rate, warming rate, and storage efficiency. Experiments were performed with six amounts of skin ranging from 7.6 × 20 cm (0.17 ft²) up to 7.6 × 120 cm (1.00 ft²).Contrary to previously published statements, when skin packaged in either of the two formats is cooled at an uncontrolled rate in a low temperature (?70 °C) mechanical refrigerator or dry-ice chest, the smaller skin dimensions cool too rapidly (up to ?24 °C min^?1), while the packets containing larger skin dimensions exhibit prolonged exothermic temperature plateaus (8–44 min), allowing the possibility of significant crystallization damage to the cells. On the other hand, controlled-rate cooling of ?1 °C min^?1 can be obtained using a temperature-feedback controlled-rate freezer along with a flat skin packet geometry. Much less control is obtained if a cylindrical skin packet geometry is used with a controlled-rate freezer.Skin processed in the flat format is capable of being warmed by water immersion about 10 times more quickly than equivalent amounts of skin processed in the rolled format. The longer warming times associated with the cylindrical package format (3.5–25 min, depending upon the amount of skin per packet) result from extended endothermic temperature plateaus in the subzero region, which have been shown to damage skin cells and reduce their subsequent viability. The short warming times (0.25–3.5 min) associated with the flat skin package format are devoid of such complications, since they are within the needed warming rate of 50 °C–70 °C min^?1.Package geometry affects the storage requirements of transplantable skin. The flat format possesses a two- to threefold advantage in storage efficiency. Capital equipment and liquid nitrogen usage for storage is drastically decreased if a flat package format is chosen.