Rules to limit snacking in front of the TV and childhood BMI status: Findings from the "Fun Families Study"

Childhood obesity is increasing at epidemic rates, and thus there is a need to target appropriate childhood behaviors that contribute to obesity. Many factors contribute to childhood weight status. The aim of this study was to look at relationships between parental rules to limit snacking while watching television and childhood weight status. The study looked at the presence of the behavior of snacking while watching television yesterday, congruence between child- and parent-reported perception of the presence of rules to limit snacking while watching television, and parent-reported frequency of children following rules to limit snacking while watching television. The outcomes were examined in a multi-ethnic population of children ages 6 to 9 years in Southeast Texas.^ This study was a cross-sectional secondary data analysis of the pilot program, Fun Families. This study examined baseline data from 202 parent-child dyads, which included both the control ( N= 101) and intervention groups (N= 101). Data were gathered using validated questions that were administered to 6-9 year old children and their primary caregiver (referred to as parent in the rest of the discussion) in Southeast Texas, between 2006 and 2008. The main study outcome was childhood weight status based on CDC BMI-for-age categories. The independent variables are (1) the presence of parental rules to limit snacking while watching television, (2) the congruence between child and parent about the presence of rules to limit snacking while watching television, and (3) the parent-reported frequency of the child following the rules to limit snacking while watching television. Chi-Square analyses were used to determine if weight status was different for (1) children who reported rules to limit snacking yesterday, (2) children who reported snacking, (3) children whose parents reported rules were present, and (4) those who had rule congruence with the parents not. Chi-Square analyses also examined if there was a difference in the presence of snacking behavior for children who reported rules, for children whose parents reported rules, and for those children who had congruence about rules. Linear regressions were used to determine if any of the studied variables predicted increased weight status or reported snacking while watching television yesterday.^ This study found that child-reported snacking yesterday was significantly different for children who reported rules (4.12, p= 0.04). Child-reported rules was significantly associated with (p= -0.14, α= 0.04) and predicted child-reported snacking yesterday (R 2 0.021, p= 0.04, t= -2.04, 95% CI -0.31, -0.01). There was statistical significant incongruence between child and parent perception about the presence of rules to limit snacking yesterday (15.06, p= 0.00). For this population, parent education level was significantly associated with child-reported rules (r= -0.16, p= 0.02), child-reported snacking yesterday (r= -0.15, p= 0.04), and parent-reported frequency of child following rules to limit snacking (r= 0.29, p= -0.01). Parent-reported speaking another language besides English at home was significantly associated with parent-reported rules (r= 0.17, p= 0.02).^ Although the studied variables did not show any significant associations or predictors for childhood weight status, the significant discord between parent and child perception about the presence of rules provides valuable information to future interventions that aim to reduce childhood weight status. Including the creation and enforcement of parental rules in interventions to reduce childhood weight status will be beneficial for future studies.^

2-methylthio-N$\sp6$-dimethylallyl modification of adenosine 37 of tRNAs in {\it Escherichia coli\/} K-12

The hypermodified, hydrophobic 2-methylthio-N$\sp6$-(dimethylallyl)-adenosine (ms${2{\cdot}6}\atop1$A) residue occurs $3\sp\prime$ to the anticodon in tRNA species that read codons beginning with U. The first step (i$\sp6$A37 formation) of this modification is catalyzed by dimethylallyl diphosphate:tRNA dimethyallyltransferase (EC 2.5.1.8), which is the product of the miaA gene. Subsequent steps were proposed to be catalyzed by MiaB and MiaC enzymes to complete the ms${2{\cdot}6}\atop1$A37 modification. The study of functions of the ms${2{\cdot}6}\atop1$A37 is very important because this modified base is one of the best candidates for a role in global control in response to environmental stress. This dissertation describes the further delineation of functions of the ms${2{\cdot}6}\atop1$A37 modification in E. coli K-12 cells. This work provides significant information on functions of tRNA modifications in E. coli cells to adapt to stressful environmental conditions. Three hypotheses were tested in this work.^ The first hypothesis tested was that non-optimal translation processes cause increased spontaneous mutagenesis by the induction of SOS response in starving cells. To test this hypothesis, I measured spontaneous mutation rates of wild type cells and various mutant strains which are defective in tRNA modification, SOS response, or oxidative damage repair. I found that the miaA mutation acts as a mutator that increased Lac$\sp+$ reversion rates and Trp$\sp+$ reversion frequencies of the wild-type cells in starving conditions. However, the lexA3(Ind)(which abolishes the induction of SOS response) mutation abolished the mutator phenotype of the miaA mutant. The recA430 mutation, not other identified SOS genes, decreased the Lac$\sp+$ reversion to a less extent than that of the lexA3(Ind) mutation. These results suggest that RecA together with another unidentified SOS gene product are responsible for the process.^ The second hypothesis tested was that MiaA protein binds to full-length tRNA$\sp{\rm Phe}$ molecules in form of a protein dimer. To test this hypothesis, three versions of the MiaA protein and seven species of tRNA substrates were purified. Binding studies by gel mobility shift assays, filter binding assays and gel filtration shift assays support the hypothesis that MiaA protein binds to full-length tRNA$\sp{\rm Phe}$ as a protein dimer but as a monomer to the anticodon stem-and-loop. These results were further supported by using steady state enzyme kinetic studies.^ The third hypothesis tested in this work was that the miaB gene in E. coli exists and is clonable. The miaB::Tn10dCm insertion mutation of Salmonella typhimurium was transduced to E. coli K-12 cells by using P$\sb1$ and P$\sb{22}$ bacteriophages. The insertion was confirmed by HPLC analyses of nucleotide profiles of miaB mutants of E. coli. The insertion mutation was cloned and DNA sequences adjacent to the transposon were sequenced. These DNA sequences were 86% identical to the f474 gene at 14.97 min chromosome of E. coli. The f474 gene was then cloned by PCR from the wild-type chromosome of E. coli. The recombinant plasmid complemented the mutant phenotype of the miaB mutant of E. coli. These results support the hypothesis that the miaB gene of E. coli exists and is clonable. In summary, functions of the ms${2{\cdot}6}\atop1$A37 modification in E. coli cells are further delineated in this work in perspectives of adaptation to stressful environmental conditions and protein:tRNA interaction. (Abstract shortened by UMI.) ^

DNA-DAMAGING REACTIONS OF NEOCARZINOSTATIN (ANTIBIOTICS, DNA REPAIR, CYTOTOXICITY)

Sensitive assays utilizing a cell-free and an intracellular system were employed to study the molecular bases of the DNA-damaging reactions of neocarzinostatin (NCS). In the cell-free DNA system, super-helical form I DNA from the bacteriophage PM2 was used as the substrate. The three forms of DNA present after treatment with NCS were separated by agarose gel electrophoresis. When NCS-damaged DNA was assayed under neutral conditions, there was a progressive decrease in the amount of surviving form I DNA and a corresponding increase in form II (nicked, relaxed circular) DNA, but very little increase in form III (linear duplex) DNA. This indicates that NCS introduces primarily single-strand breaks. However later studies showed that there were some site-specific double-strand breaks mediated by NCS on PM2 DNA. Seven such specific sites were mapped on the PM2 genome. When the damage was assayed under nondenaturing alkaline conditions or with the apurinic/apyrimidinic endonuclease IV, there was a slightly greater decrease in the amount of surviving form I DNA compared with neutral conditions indicating the presence of some alkali-labile sites.^ NCS-mediated DNA damage and repair were examined with cultured Chinese hamster ovary (CHO) cells using either alkaline elution for analysis of single-strand breaks or neutral elution for analysis of double-strand breaks. Most of the strand breaks introduced by NCS were capable of being rejoined. However, there was a small amount of residual DNA damage remaining unrejoined at 24-hr after removal of the drug. The amount of residual DNA damage was higher in a CHO mutant cell line (EM9) having a higher sensitivity to killing by NCS than its parental strain (AA8). Other lesions, DNA-protein complexes and alkali-labile sites, were detected after NCS treatment but they constituted only a small fraction of the DNA damage.^ Based on the above information, it can be postulated that NCS introduces some very lethal DNA damage. It is likely that the lethal lesions are a subset of the total DNA lesions representing the residual DNA damage. This DNA damage may be composed of site-specific, unrejoinable double-strand breaks and are thus the primary lesion leading to NCS-mediated lethality.^