Adult Inception of Ketogenic Diet Therapy Increases Sleep during the Dark Cycle in C57BL/6J Wild Type and Fragile X Mice.

Sleep problems are a significant phenotype in children with fragile X syndrome. Our prior work assessed sleep-wake cycles in Fmr1KO male mice and wild type (WT) littermate controls in response to ketogenic diet therapy where mice were treated from weaning (postnatal day 18) through study completion (5-6 months of age). A potentially confounding issue with commencing treatment during an active period of growth is the significant reduction in weight gain in response to the ketogenic diet. The aim here was to employ sleep electroencephalography (EEG) to assess sleep-wake cycles in mice in response to the Fmr1 genotype and a ketogenic diet, with treatment starting at postnatal day 95. EEG results were compared with prior sleep outcomes to determine if the later intervention was efficacious, as well as with published rest-activity patterns to determine if actigraphy is a viable surrogate for sleep EEG. The data replicated findings that Fmr1KO mice exhibit sleep-wake patterns similar to wild type littermates during the dark cycle when maintained on a control purified-ingredient diet but revealed a genotype-specific difference during hours 4-6 of the light cycle of the increased wake (decreased sleep and NREM) state in Fmr1KO mice. Treatment with a high-fat, low-carbohydrate ketogenic diet increased the percentage of NREM sleep in both wild type and Fmr1KO mice during the dark cycle. Differences in sleep microstructure (length of wake bouts) supported the altered sleep states in response to ketogenic diet. Commencing ketogenic diet treatment in adulthood resulted in a 15% (WT) and 8.6% (Fmr1KO) decrease in body weight after 28 days of treatment, but not the severe reduction in body weight associated with starting treatment at weaning. We conclude that the lack of evidence for improved sleep during the light cycle (mouse sleep time) in Fmr1KO mice in response to ketogenic diet therapy in two studies suggests that ketogenic diet may not be beneficial in treating sleep problems associated with fragile X and that actigraphy is not a reliable surrogate for sleep EEG in mice.

Effects of Soy Protein Isolate on Fragile X Phenotypes in Mice.

Obesity is a pediatric epidemic that is more prevalent in children with developmental disabilities. We hypothesize that soy protein-based diets increase weight gain and alter neurobehavioral outcomes. Our objective herein was to test matched casein- and soy protein-based purified ingredient diets in a mouse model of fragile X syndrome, Fmr1KO mice. The experimental methods included assessment of growth; 24-7 activity levels; motor coordination; learning and memory; blood-based amino acid, phytoestrogen and glucose levels; and organ weights. The primary outcome measure was body weight. We find increased body weight in male Fmr1KO from postnatal day 6 (P6) to P224, male wild type (WT) from P32-P39, female Fmr1KO from P6-P18 and P168-P224, and female Fmr1HET from P9-P18 as a function of soy. Activity at the beginning of the light and dark cycles increased in female Fmr1HET and Fmr1KO mice fed soy. We did not find significant differences in rotarod or passive avoidance behavior as a function of genotype or diet. Several blood-based amino acids and phytoestrogens were significantly altered in response to soy. Liver weight was increased in WT and adipose tissue in Fmr1KO mice fed soy. Activity levels at the beginning of the light cycle and testes weight were greater in Fmr1KO versus WT males irrespective of diet. DEXA analysis at 8-months-old indicated increased fat mass and total body area in Fmr1KO females and lean mass and bone mineral density in Fmr1KO males fed soy. Overall, dietary consumption of soy protein isolate by C57BL/6J mice caused increased growth, which could be attributed to increased lean mass in males and fat mass in females. There were sex-specific differences with more pronounced effects in Fmr1KO versus WT and in males versus females.

Fecal indicator bacteria (FIB) levels during dry weather from Southern California reference streams.

High levels of fecal indicator bacteria (FIB) in surface waters is a common problem in urban areas that often leads to impairment of beneficial uses such as swimming. Once impaired, common management and regulatory solutions include development of total maximum daily loads (TMDLs) and other water quality management plans. A critical element of these plans is establishment of a "reference" level of exceedances against which to assess management goals and TMDL compliance. The goal of this study was to provide information on indicator bacteria contributions from natural streams in undeveloped catchments throughout southern California during dry weather, non-storm conditions. To help establish a regional reference data set, bacteria levels [i.e. Escherichia coli (E. coli), enterococci and total coliforms] were measured from 15 unimpaired streams in 11 southern California watersheds weekly for one full year. Concentrations measured from reference areas were typically between one to two orders of magnitude lower than levels found in developed watersheds. Nearly 82% of the time, samples did not exceed daily and monthly bacterial indicator thresholds. E. coli had the lowest daily percent exceedance (1.5%). A total of 13.7% of enterococci exceeded daily thresholds. Indicator bacteria levels fluctuated seasonally with an average of 79% of both enterococci and total coliforms exceedances occurring during summer months (June to August). Temperature, at all sites, explained about one-half the variation in total coliforms density suggesting that stream temperatures regulated bacterial populations. Accounting for natural background levels will allow for management targets that are more reflective of the contributions from natural sources.

How effective has the Clean Water Act been at reducing pollutant mass emissions to the Southern California Bight over the past 35 years?

The Clean Water Act (CWA) has regulated discharges of contaminants since 1972. However, evaluations of the CWA's effectiveness at improving regional water quality are lacking, primarily because integration of monitoring data from multiple dischargers to assess cumulative effects is not required. A rare opportunity exists to assess CWA effectiveness by integrating mass emissions data from all major sources of contaminants to the Southern California Bight from 1971 to 2000. While the coastal population grew by 56% and total effluent volume increased 31% since 1971, mass emissions of nearly all constituents decreased since passage of the CWA, most by greater than 65%. Publicly owned treatment works were the dominant point source of many contaminants, but also accounted for the greatest reductions in pollutant discharge since 1971. As point source treatment has improved, the relative contribution of non-point sources, such as storm water runoff has increased. Despite the increased importance of storm water discharges, regional monitoring and data compilation of this source is lacking, making it difficult to accurately assess trends in non-point source discharge.