Nucleic acid relationships among Acholeplasma species.

3H-labeled Acholeplasma DNA probes were generated in vitro by the nick-translation method and used to determine the nucleotide sequence homology among the type strains of the eight currently recognized species of Acholeplasma. Very little nucleotide sequence homology (less than or equal to 18%) was found among the eight species, with heteroduplexes showing at least 12% or more mismatching as determined by thermal elution midpoints. The small amount of nucleotide sequence homology among the eight species indicates that these species are quite distinct and are not closely related to each other genomically.     

Quantifying Biomass Changes of Single CD8+ T Cells during Antigen Specific Cytotoxicity

Existing approaches that quantify cytotoxic T cell responses rely on bulk or surrogate measurements which impede the direct identification of single activated T cells of interest. Single cell microscopy or flow cytometry methodologies typically rely on fluorescent labeling, which limits applicability to primary cells such as human derived T lymphocytes. Here, we introduce a quantitative method to track single T lymphocyte mediated cytotoxic events within a mixed population of cells using live cell interferometry (LCI), a label-free microscopy technique that maintains cell viability. LCI quantifies the mass distribution within individual cells by measuring the phase shift caused by the interaction of light with intracellular biomass. Using LCI, we imaged cytotoxic T cells killing cognate target cells. In addition to a characteristic target cell mass decrease of 20–60% over 1–4 h following attack by a T cell, there was a significant 4-fold increase in T cell mass accumulation rate at the start of the cytotoxic event and a 2–3 fold increase in T cell mass relative to the mass of unresponsive T cells. Direct, label-free measurement of CD8+ T and target cell mass changes provides a kinetic, quantitative assessment of T cell activation and a relatively rapid approach to identify specific, activated patient-derived T cells for applications in cancer immunotherapy.     

Isomeric lipid signatures reveal compartmentalized fatty acid metabolism in cancer

The cellular energy and biomass demands of cancer drive a complex dynamic between uptake of extracellular FAs and their de novo synthesis. Given that oxidation of de novo synthesized FAs for energy would result in net-energy loss, there is an implication that FAs from these two sources must have distinct metabolic fates; however, hitherto, all FAs have been considered part of a common pool. To probe potential metabolic partitioning of cellular FAs, cancer cells were supplemented with stable isotope-labeled FAs. Structural analysis of the resulting glycerophospholipids revealed that labeled FAs from uptake were largely incorporated to canonical (sn-) positions on the glycerol backbone. Surprisingly, labeled FA uptake also disrupted canonical isomer patterns of the unlabeled lipidome and induced repartitioning of n-3 and n-6 PUFAs into glycerophospholipid classes. These structural changes support the existence of differences in the metabolic fates of FAs derived from uptake or de novo sources and demonstrate unique signaling and remodeling behaviors usually hidden from conventional lipidomics.     

Identification and functional analysis of NOL7 nuclear and nucleolar localization signals

Background  

NOL7 is a candidate tumor suppressor that localizes to a chromosomal region 6p23. This locus is frequently lost in a number of malignancies, and consistent loss of NOL7 through loss of heterozygosity and decreased mRNA and protein expression has been observed in tumors and cell lines. Reintroduction of NOL7 into cells resulted in significant suppression of in vivo tumor growth and modulation of the angiogenic phenotype. Further, NOL7 was observed to localize to the nucleus and nucleolus of cells. However, the mechanisms regulating its subcellular localization have not been elucidated.     

Food sharing: a model of manipulation by harassment

Most analyses of food-sharing behavior invoke complex explanationssuch as indirect and delayed benefits for sharing via kin selectionand reciprocal altruism. However, food sharing can be a moregeneral phenomenon accounted for by more parsimonious, mutualisticexplanations. We propose a game theoretical model of a generalsharing situation in which food owners share because it is in their own self-interest—they avoid high costs associatedwith beggar harassment. When beggars harass, owners may benefitfrom sharing part of the food if their consumption rate islow relative to the rate of cost accrual. Our model predictsthat harassment can be a profitable strategy for beggars if they reap some direct benefits from harassing other than sharedfood (such as picking up scraps). Therefore, beggars may manipulatethe owner's fitness payoffs in such a way as to make sharingmutualistic.     

Ultrastructural changes in isolated rat hepatocytes exposed to different CCl4 concentrations

Ultrastructural data are presented on time-course changes in isolated rat hepatocyte suspensions exposed either to 1.2 or 1.8 mM CCl4 for up to 1 h. The subcellular changes at the lower concentration, but not the higher, are shown to closely parallel those reported to occur in rat hepatocytes following ingestion of CCl4.     

Solution Radioactivated by Hadron Radiation Can Increase Sister Chromatid Exchanges

When energetic particles irradiate matter, it becomes activated by nuclear reactions. Radioactivation induced cellular effects are not clearly understood, but it could be a part of bystander effects. This investigation is aimed at understanding the biological effects from radioactivation in solution induced by hadron radiation. Water or phosphate buffered saline was activated by being exposed to hadron radiation including protons, carbon- and iron-ions. 1 mL of radioactivated solution was transferred to flasks with Chinese hamster ovary (CHO) cells cultured in 5 mL of complete media. The induction of sister chromatid exchanges (SCE) was used to observe any increase in DNA damage responses. The energy spectrum and the half-lives of the radioactivation were analyzed by NaI scintillation detector in order to identify generated radionuclides. In the radioactivated solution, 511 keV gamma-rays were observed, and their half-lives were approximately 2 min, 10 min, and 20 min. They respectively correspond to the beta+ decay of ¹⁵O, ¹³N, and ¹¹C. The SCE frequencies in CHO cells increased depending on the amount of radioactivation in the solution. These were suppressed with a 2-hour delayed solution transfer or pretreatment with dimethyl sulfoxide (DMSO). Our results suggest that the SCE induction by radioactivated solution was mediated by free radicals produced by the annihilated gamma-rays. Since the SCE induction and DMSO modulation are also reported in radiation-induced bystander effects, our results imply that radioactivation of the solution may have some contribution to the bystander effects from hadron radiation. Further investigations are required to assess if radioactivation effects would attribute an additional level of cancer risk of the hadron radiation therapy itself.     

Natal recruitment and adult retention in a population of nine-banded armadillos

Nine-banded armadillosDasypus novemcinctus Linnaeus, 1758 are interesting in part because (a) they give birth to litters of genetically identical quadruplets, and (b) the species’ range has expanded rapidly throughout the southern United States during the last 150 years, suggesting substantial dispersal of individuals. Using data from 7 field seasons between 1992 and 1999, we examined the extent of juvenile recruitment and retention of adults in a population of armadillos from northern Florida. There were no sex differences in the likelihood of recruitment or most attributes of male and female recruits at any age. In the few cases where more than one littermate was recruited into the population, siblings were significantly more widely dispersed as adults than they were as juveniles, thus limiting opportunities for interaction among clonal siblings. There was some evidence that recruits ranged more widely than other individuals, suggesting recruits may have been searching for suitable sites to establish themselves. Recruits were heavier than non-recruits as both juveniles and yearlings, which may have aided in establishing a home range, but recruits were lighter than other animals as adults. Overall, slightly less than 50% of armadillos first captured as adults were never seen in a subsequent year, suggesting these individuals may have been transients. However, some adults remained in the population for multiple years, moving very little from the area where they were first sighted. As with recruits, there were no sex differences in the likelihood of adults being retained in the population nor in the attributes of retained males and females. Retained animals exhibited more extensive anatomical damage and moved farther between successive sightings within years than did non-residents. Adults were more likely to be retained in the population than juveniles were to be recruited, and retained adults were older, heavier, and exhibited more extensive anatomical damage than did recruits. Our data seem to indicate a population characterized by limited recruitment of juveniles (particularly of clonemates) and an adult population exhibiting considerable turnover from year to year, but with a core of individuals who are long-term residents.