Inhibition of L-type amino acid transport with non-physiological amino acids in the Pahenu2 mouse model of phenylketonuria

Phenylketonuria, an autosomal recessive Mendelian disorder, is one of the most common inborn errors of metabolism. Although currently treated by diet, many suboptimal outcomes occur for patients. Neuropathological outcomes include cognitive loss, white matter abnormalities, and hypo- or demyelination, resulting from high concentrations and/or fluctuating levels of phenylalanine. High phenylalanine can also result in competitive exclusion of other large neutral amino acids from the brain, including tyrosine and tryptophan (essential precursors of dopamine and serotonin). This competition occurs at the blood brain barrier, where the L-type amino acid transporter, LAT1, selectively facilitates entry of large neutral amino acids. The hypothesis of these studies is that certain non-physiological amino acids (NPAA; DL-norleucine (NL), 2-aminonorbornane (NB; 2-aminobicyclo-(2,1,1)-heptane-2-carboxylic acid), α-aminoisobutyrate (AIB), and α-methyl-aminoisobutyrate (MAIB)) would competitively inhibit LAT1 transport of phenylalanine (Phe) at the blood-brain barrier interface. To test this hypothesis, Pah-/- mice (n=5, mixed gender; Pah+/-(n=5) as controls) were fed either 5% NL, 0.5% NB, 5% AIB or 3% MAIB (w/w 18% protein mouse chow) for 3 weeks. Outcome measurements included food intake, body weight, brain LNAAs, and brain monoamines measured via LCMS/MS or HPLC. Brain Phe values at sacrifice were significantly reduced for NL, NB, and MAIB, verifying the hypothesis that these NPAAs could inhibit Phe trafficking into the brain. However, concomitant reductions in tyrosine and methionine occurred at the concentrations employed. Blood Phe levels were not altered indicating no effect of NPAA competitors in the gut. Brain NL and NB levels, measured with HPLC, verified both uptake and transport of NPAAs. Although believed predominantly unmetabolized, NL feeding significantly increased blood urea nitrogen. Pah-/-disturbances of monoamine metabolism were exacerbated by NPAA intervention, primarily with NB (the prototypical LAT inhibitor). To achieve the overarching goal of using NPAAs to stabilize Phe transport levels into the brain, a specific Phe-reducing combination and concentration of NPAAs must be found. Our studies represent the first in vivo use of NL, NB and MAIB in Pah-/- mice, and provide proof-of-principle for further characterization of these LAT inhibitors. Our data is the first to document an effect of MAIB, a specific system A transport inhibitor, on large neutral amino acid transport.

Triggering of volcanic activity by large earthquakes

Statistical analyses of temporal relationships between large earthquakes and volcanic eruptions suggest seismic waves may trigger eruptions even over great (>1000 km) distances, although the causative mechanism is not well constrained. In this study the relationship between large earthquakes and subtle changes in volcanic activity was investigated in order to gain greater insight into the relationship between dynamic stresses propagated by surface waves and volcanic response. Daily measurements from the Ozone Monitoring Instrument (OMI), onboard the Aura satellite, provide constraints on volcanic sulfur-dioxide (SO2) emission rates as a measure of subtle changes in activity. Time series of SO2 emission rates were produced from OMI data for thirteen persistently active volcanoes from 1 October 2004 to 30 September 2010. In order to quantify the affect of earthquakes at teleseismic distances, we modeled surface-wave amplitudes from the source mechanisms of moment magnitude (Mw) ≥7 earthquakes, and calculated the Peak Dynamic Stress (PDS). We assessed the influence of earthquakes on volcanic activity in two ways: 1) by identifying increases in the SO2 time series data and looking for causative earthquakes and 2) by examining the average emission rate before and after each earthquake. In the first, the SO2 time series for each volcano was used to calculate a baseline threshold for comparison with post-earthquake emission. Next, we generated a catalog of responses based on sustained SO2 emission increases above this baseline. Delay times between each SO2 response and each prior earthquake were analyzed using both the actual earthquake catalog, and a randomly generated catalog of earthquakes. This process was repeated for each volcano. Despite varying multiple parameters, this analysis did not demonstrate a clear relationship between earthquake-generated PDS and SO2 emission. However, the second analysis, which was based on the occurrence of large earthquakes indicated a response at most volcanoes. Using the PDS calculations as a filtering criterion for the earthquake catalog, the SO2 mass for each volcano was analyzed in 28-day windows centered on the earthquake origin time. If the average SO2 mass after the earthquake was greater than an arbitrary percentage of pre-earthquake mass, we identified the volcano as having a response to the event. This window analysis provided insight on what type of volcanic activity is more susceptible to triggering by dynamic stress. The volcanoes with very open systems included in this study, Ambrym, Gaua, Villarrica, Erta Ale and, Turrialba, showed a clear response to dynamic stress while the volcanoes with more closed systems, Merapi, Semeru, Fuego, Pacaya, and Bagana, showed no response.