Soluble Reactive Phosphorus Transport and Retention in Tropical, Rainforest Streams Draining a Volcanic and Geothermally Active Landscape in Costa Rica. : Long-Term Concentration Patterns, Pore Water Environment and Response to ENSO Events

Soluble reactive phosphorus (SRP) transport/retention was determined at four sites in three rainforest streams draining La Selva Biological Station, Costa Rica. La Selva is located at the base of the last remaining intact rainforest transect from 30 m above sea level to 3000 m along the entire Caribbean slope of Central America. Steam SRP levels can be naturally high there due to regional, geothermal groundwater discharged at ambient temperature. Monitoring since 1988 has revealed distinctive long-term differences in background SRP and total P (TP) for three streams in close proximity, and identified the impact of ENSO (El Nino Southern Oscillation) events on SRP-enriched reaches. Mean interannual SRP concentrations (± standard deviation) were 89 ± 53μg/l in the Salto (1988–1996), 21 ± 39μg/l in the Pantano (1988–1998), and 26 ± 35μg/l in the Sabalo (1988–1996). After January, 1997 the separate upland-lowland contributions to discharge and SRP load were determined monthly in the Salto. SRP in Upper Salto was low (19 ± 8μg/l, 1997–2002) until enriched at␣the upland-lowland transition by regional groundwater. Mean SRP concentration in Lower␣Salto (108 ± 104μg/l) was typically highest February–April, the driest months, and lowest July–September, the wettest. SRP concentration was positively correlated to the inverse of discharge in Lower Salto when ENSO data were omitted (1992 and 1998–1999), but not in the Upper Salto, Pantano, or Sabalo. TP was positively correlated to the inverse of discharge in all three streams when ENSO data were omitted. High SRP springs and seeps along the Lower Salto contributed 36% of discharge but 85% of SRP export 1997–2001. Annual SRP flux from the total Salto watershed (1997–2001) averaged 2.9 kg/ha year, but only 0.6 kg/ha year from the Upper Salto. A dye tracer injection showed that pore water environments were distinctly different between Upper and Lower Salto. Upper Salto had high surface water–pore water exchange, high dissolved oxygen, low SRP, and low conductivity similar to surface water, and Lower Salto had low surface water–pore water exchange, low dissolved oxygen, high SRP, and high conductivity reflecting geothermal groundwater influence. SRP export from the Salto was controlled by regional groundwater transfer, which in similar volcanic settings could be a significant P source. However, ENSO events modified the SRP concentration in the Salto suggesting that long-term monitoring is required to understand underlying SRP dynamics and P flux to downstream communities.     

Hydrogen sulfide as an oxygen sensor in trout gill chemoreceptors

O2 chemoreceptors elicit cardiorespiratory reflexes in all vertebrates, but consensus on O2-sensing signal transduction mechanism(s) is lacking. We recently proposed that hydrogen sulfide (H2S) metabolism is involved in O2 sensing in vascular smooth muscle. Here, we examined the possibility that H2S is an O2 sensor in trout chemoreceptors where the first pair of gills is a primary site of aquatic O2 sensing and the homolog of the mammalian carotid body. Intrabuccal injection of H2S in unanesthetized trout produced a dose-dependent bradycardia and increased ventilatory frequency and amplitude similar to the hypoxic response. Removal of the first, but not second, pair of gills significantly inhibited H2S-mediated bradycardia, consistent with the loss of aquatic chemoreceptors. mRNA for H2S-synthesizing enzymes, cystathionine beta-synthase and cystathionine gamma-lyase, was present in branchial tissue. Homogenized gills produced H2S enzymatically, and H2S production was inhibited by O2, whereas mitochondrial H2S consumption was O2 dependent. Ambient hypoxia did not affect plasma H2S in unanesthetized trout, but produced a PO2-dependent increase in a sulfide moiety suggestive of increased H2S production. In isolated zebrafish neuroepithelial cells, the putative chemoreceptive cells of fish, both hypoxia and H2S, produced a similar approximately 10-mV depolarization. These studies are consistent with H2S involvement in O2 sensing/signal transduction pathway(s) in chemoreceptive cells, as previously demonstrated in vascular smooth muscle. This novel mechanism, whereby H2S concentration ([H2S]) is governed by the balance between constitutive production and oxidation, tightly couples tissue [H2S] to PO2 and may provide an exquisitely sensitive, yet simple, O2 sensor in a variety of tissues.     

Alterations in carbohydrate metabolism and its regulation in PPARalpha null mouse hearts

Although a shift from fatty acids (FAs) to carbohydrates (CHOs) is considered beneficial for the diseased heart, it is unclear why subjects with FA beta-oxidation defects are prone to cardiac decompensation under stress conditions. The present study investigated potential alterations in the myocardial utilization of CHOs for energy production and anaplerosis in 12-wk-old peroxisome proliferator-activating receptor-alpha (PPARalpha) null mice (a model of FA beta-oxidation defects). Carbon-13 methodology was used to assess substrate flux through energy-yielding pathways in hearts perfused ex vivo at two workloads with a physiological substrate mixture mimicking the fed state, and real-time RT-quantitative polymerase chain reaction was used to document the expression of selected metabolic genes. When compared with that from control C57BL/6 mice, isolated working hearts from PPARalpha null mice displayed an impaired capacity to withstand a rise in preload (mimicking an increased venous return as it occurs during exercise) as reflected by a 20% decline in the aortic flow rate. At the metabolic level, beyond the expected shift from FA (5-fold down) to CHO (1.5-fold up; P < 0.001) at both preloads, PPARalpha null hearts also displayed 1) a significantly greater contribution of exogenous lactate and glucose and/or glycogen (2-fold up) to endogenous pyruvate formation, whereas that of exogenous pyruvate remained unchanged and 2) marginal alterations in citric acid cycle-related parameters. The lactate production rate was the only measured parameter that was affected differently by preloads in control and PPARalpha null mouse hearts, suggesting a restricted reserve for the latter hearts to enhance glycolysis when the energy demand is increased. Alterations in the expression of some glycolysis-related genes suggest potential mechanisms involved in this defective CHO metabolism. Collectively, our data highlight the importance of metabolic alterations in CHO metabolism associated with FA oxidation defects as a factor that may predispose the heart to decompensation under stress conditions even in the fed state.     

Increased Dopaminergic Neuron Sensitivity to 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine (MPTP) in Transgenic Mice Expressing Mutant A53T α-Synuclein

Familial Parkinson’s disease (PD) has been linked to point mutations and duplication of the α-synuclein gene and mutant α-synuclein expression increases the vulnerability of neurons to exogenous insults. In this study, we analyzed the levels of dopamine and its metabolites in the olfactory bulb (OB), and nigrostriatal regions of transgenic mice expressing human, mutant A53T α-synuclein (α-syn tg) and their non-transgenic (ntg) littermates using a sub-toxic, moderate dose of MPTP to determine if mutant human α-synuclein sensitizes the central dopaminergic systems to oxidative stress. We observed that after a single, sub-lethal MPTP injection, dopamine levels were reduced in striatum and SN in both the α-syn tg and ntg mice. In the olfactory bulb, a region usually resistant to MPTP toxicity, levels were reduced only in the α-syn tg mice. In addition, we identified a significant increase in dopamine metabolism in the α-syn transgenic, but not ntg mice. Finally, MPTP treatment of α-syn tg mice was associated with a marked elevation in the oxidative product, 3-nitrotyrosine that co-migrated with α-synuclein. Cumulatively, the data support the hypothesis that mutant α-synuclein sensitizes dopaminergic neurons to neurotoxic insults and is associated with greater oxidative stress. The α-syn tg line is therefore useful to study the genetic and environmental inter-relationship in PD.     

Identification of an interleukin-1 beta binding protein in human plasma

A covalent cross-linking technique was used to bind iodinated interleukin-1 (IL1) alpha and beta to plasma proteins. One specific IL1 beta binding protein was observed, that when cross-linked to ¹²⁵I-ILl beta migrated to approximately 60 kDa on SDS-PAGE. The protein did not bind IL1 alpha. The 43 -kDa protein was partially purified using a wheat germ agglutinin affinity column. The isolated factor again specifically bound IL1 beta, and appeared to consist of single chain glycoprotein. The protein was heat stable and had a rapid association time with IL1 beta. This protein may be an important carrier molecule for IL1 beta in vivo.