Studies on the control of mitotic activity

Summary A tetraploid cell population was produced in the primary root meristem of Pisum sativum by one-half hour treatments with various concentrations of colchicine. The tetraploid population so produced was found to be reasonably synchronous in its passage through successive mitotic cycles with the degree of synchrony being more or less proportional to the concentration of colchicine used. The average time between mitoses appears to be of the order of 12 hours which agrees well with previous estimates. Treatments of roots containing tetraploid populations with 2.52 × 10^–5 M. actidione for 15 minutes were used to demonstrate the possibility of using the system for studies on the differential susceptibility of cells at different stages of the mitotic cycle.This work was carried out under contract number RG-4835 of the National Institutes of Health, United States Public Health Service, and an Institutional Grant from the American Cancer Society.Contribution number 59-26 of the Department of Botany and Plant Pathology, Michigan State University, East Lansing, Michigan.Predoctoral Fellow (CF-9871) of the National Cancer Institute, United States Public Health Service     

Capacity of intact proteins to bind to MHC class II molecules

Here we have demonstrated that denatured, but not native protein antigens can interact with Ia molecules. Thus, the failure of native antigens to be recognized as such by T cells appears to be at least in part due to a deficient antigen/Ia interaction. These results also support previous observations that some T cells can recognize denatured antigens without a further processing requirement. Moreover, a striking correlation was observed between the in vitro binding pattern of denatured proteins and the pattern of restriction of T cell responses elicited by immunization with the native antigen, raising the possibility that an unfolding step may actually occur early during in vivo processing and influence the final outcome of Ia-restricted T cell responses.     

An increase of acidic isoform of catalase in red blood cells from HIV(+) population

A systemic oxidative stress of HIV (+) individuals has been recognized from a low glutathione level and a high level of inflammatory cytokines such as TNF. Previously, we demonstrated that the catalase enzyme activity in HIV (+) population is significantly altered depending on the cell types; the level was significantly high in red blood cells while the enzymes in white blood cells were remarkably low (Res Commun Subs Abuse 16: 161–176, 1995). In this study, we further characterized the difference in RBC catalase molecules between HIV (+) and control population. We have found that RBC from HIV (+) population, whether they were asymptomatic or symptomatic, contained a significantly elevated catalase protein accompanied by the enzyme activities, and that the majority of the elevated protein were acidic pl of the molecules with an identical subunit mass of approximately 60 KDa. These results suggest that catalase is induced prior to and/or during erythroid differentiation lineage in HIV (+) population as a somatic defense to respond and compensate for a systemic oxidative stress and for an anemic condition. (Mol Cell Biochem 165: 77–81, 1996)     

Partial resistance to late blight (Phytophthora infestans) in hybrid progenies of four South American Solanum species crossed with diploid S. tuberosum

Resistant genotypes of the diploid tuber-bearing South American species Solarium arnezii x hondelmannii, S. berthaultii, S. leptophyes and S. microdontum were crossed with three diploid genotypes of S. tuberosum that varied in resistance and maturity type. The progenies were field tested for 2 years for resistance to a complex race of Phytophthora infestans. A wealth of genetic variation for resistance was found in most of the progenies. At least two susceptibility groups could be distinguished in some progenies of S. microdontum. This could be explained by the presence of several major resistance genes in the wild parent and, unexpectedly, in the susceptible parent SH 82-44-111. In most of the wild parents and in the susceptible parent SH 77-114-2988 there appeared to be minor resistance genes. General combining ability effects were predominant; small specific combining ability effects were detected in some crosses of S. microdontum. Gene action appeared dominant in some crosses.     

Advantages of distinguishing the active fraction in bacterioplankton assemblages: some examples

The difficulty of distinguishing between active and dormant or dead bacterial cells is an important problem for the aquatic microbiologist.Active cells can be detected under the microscope by the presence of an intact electron transport system able to reduce the colourless INT [2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride] to an optically dense intracellular deposit.An improvement of this method has been applied to Lake Geneva and to a fish pond in the Ivory Coast. The portion of INT-reducing bacterial cells ranged from 1 to 71%, depending on place, depth, season and time of the day. In all cases bacterial activity, determined by uptake of ³H Thymidine or ¹⁴C glucose, and frequency of dividing cells were better correlated with the number of INT-reducing cells than with the total number of cells. This means that counts of cells able to reduce INT have a better metabolic significance than total cell counts. Some examples are developed which show the advantages of applying this method in cases where it is useful to distinguish active cells in a bacterial assemblage.     

An index approach to assess nitrogen losses to the environment

Nitrogen (N) losses from agriculture are negatively impacting groundwater, air, and surface water quality. New tools are needed to quickly assess these losses and provide nutrient managers and conservationists with effective tools to assess the effects of current and alternative management practices on N loss pathways. A new N-Index tool was developed in spreadsheet format, allowing prompt assessments of management practices on agricultural N losses. The N-Index tool was compared with experimental field data and shown to estimate the effects of management practices on N loss pathways (probability, P < 0.001). The N-Index correctly assessed the nitrate nitrogen (NO₃-N) leaching losses when tested against measured NO₃-N leaching data and atmospheric N losses collected over multiple years (annual basis) and locations. The N-Index tool was developed with international cooperation from several countries and there is potential to use this tool at the international level.     

Influence of Leptin on Changes in Body Fat during Growth in African American and White Children

Objective: The aim of this study was to determine whether initial levels or temporal changes in fasting leptin were associated with longitudinal changes in body‐fat mass in children. Research Methods and Procedures: The study group consisted of 85 children (42 white and 43 African American) with a mean initial age of 8.1 ± 0.1 years. The children had between three and six annual visits for repeated measurements of body composition by DXA and fasting leptin level. Fat mass and fasting leptin level were not normally distributed and were log‐transformed. Data were analyzed using SAS Proc mixed growth models, with log fat as the dependent variable. Results: Initial leptin level was a significant predictor of the change in fat mass over time (p < 0.0001), with high initial leptin levels resulting in increased fat gain, independent of initial fat levels. This relationship remained significant when the data were analyzed separately by race (whites, p < 0.0001; African Americans, p = 0.008). The relationship between the initial level of leptin and the change in fat mass was not modified by race, sex, or Tanner Stage. The rate of change in leptin during the study was significantly related to the rate of change in fat mass in African Americans (p = 0.008) but not in whites (p = 0.490). Discussion: In conclusion, high fasting leptin level at the start of the study was significantly associated with increasing fat mass in this cohort, indicating that the children may be developing resistance to the effects of leptin.     

Growth of Visceral Fat,Subcutaneous Abdominal Fat,and Total Body Fat in Children

Objective: To examine the patterns of growth of visceral fat, subcutaneous abdominal fat, and total body fat over a 3‐ to 5‐year period in white and African American children. Research Methods and Procedures: Children (mean age: 8.1 ± 1.6 years at baseline) were recruited from Birmingham, Alabama, and those with three or more repeated annual measurements were included in the analysis (N = 138 children and 601 observations). Abdominal adipose tissue (visceral and subcutaneous) was measured using computed tomography. Total body fat and lean tissue mass were measured by DXA. Random growth curve modeling was performed to estimate growth rates of the different body fat compartments. Results: Visceral fat and total body fat both exhibited significant growth effects before and after adjusting for subcutaneous abdominal fat and lean tissue mass, respectively, and for gender, race, and baseline age (5.2 ± 2.2 cm²/yr and 1.9 ± 0.8 kg/yr, respectively). After adjusting for total body fat, the growth of subcutaneous abdominal fat was not significant. Whites showed a higher visceral fat growth than did African Americans (difference: 1.9 ± 0.8 cm²/yr), but there was no ethnic difference for growth of subcutaneous abdominal fat or total body fat. There were no gender differences found for any of the growth rates. Discussion: Growth of visceral fat remained significant after adjusting for growth of subcutaneous abdominal fat, implying that the acquisition of the two abdominal fat compartments may involve different physiologic mechanisms. In contrast, growth of subcutaneous abdominal fat was explained by growth in total body fat, suggesting that subcutaneous fat may not be preferentially deposited in the abdominal area during this phase of growth. Finally, significantly higher growth of visceral fat in white compared with African American children is consistent with cross‐sectional findings.