Genetic deciphering of the antagonistic activities of the melanin-concentrating hormone and melanocortin pathways in skin pigmentation

The genetic origin of human skin pigmentation remains an open question in biology. Several skin disorders and diseases originate from mutations in conserved pigmentation genes, including albinism, vitiligo, and melanoma. Teleosts possess the capacity to modify their pigmentation to adapt to their environmental background to avoid predators. This background adaptation occurs through melanosome aggregation (white background) or dispersion (black background) in melanocytes. These mechanisms are largely regulated by melanin-concentrating hormone (MCH) and α-melanocyte–stimulating hormone (α-MSH), two hypothalamic neuropeptides also involved in mammalian skin pigmentation. Despite evidence that the exogenous application of MCH peptides induces melanosome aggregation, it is not known if the MCH system is physiologically responsible for background adaptation. In zebrafish, we identify that MCH neurons target the pituitary gland-blood vessel portal and that endogenous MCH peptide expression regulates melanin concentration for background adaptation. We demonstrate that this effect is mediated by MCH receptor 2 (Mchr2) but not Mchr1a/b. mchr2 knock-out fish cannot adapt to a white background, providing the first genetic demonstration that MCH signaling is physiologically required to control skin pigmentation. mchr2 phenotype can be rescued in adult fish by knocking-out pomc, the gene coding for the precursor of α-MSH, demonstrating the relevance of the antagonistic activity between MCH and α-MSH in the control of melanosome organization. Interestingly, MCH receptor is also expressed in human melanocytes, thus a similar antagonistic activity regulating skin pigmentation may be conserved during evolution, and the dysregulation of these pathways is significant to our understanding of human skin disorders and cancers.     

Vitamin C prevents hyperoxia-mediated vasoconstriction and impairment of endothelium-dependent vasodilation

High arterial blood oxygen tension increases vascular resistance, possibly related to an interaction between reactive oxygen species and endothelium-derived vasoactive factors. Vitamin C is a potent antioxidant capable of reversing endothelial dysfunction due to increased oxidant stress. We tested the hypotheses that hyperoxic vasoconstriction would be prevented by vitamin C, and that acetylcholine-mediated vasodilation would be blunted by hyperoxia and restored by vitamin C. Venous occlusion strain gauge plethysmography was used to measure forearm blood flow (FBF) in 11 healthy subjects and 15 congestive heart failure (CHF) patients, a population characterized by endothelial dysfunction and oxidative stress. The effect of hyperoxia on FBF and derived forearm vascular resistance (FVR) at rest and in response to intra-arterial acetylcholine was recorded. In both healthy subjects and CHF patients, hyperoxia-mediated increases in basal FVR were prevented by the coinfusion of vitamin C. In healthy subjects, hyperoxia impaired the acetylcholine-mediated increase in FBF, an effect also prevented by vitamin C. In contrast, hyperoxia had no effect on verapamil-mediated increases in FBF. In CHF patients, hyperoxia did not affect FBF responses to acetylcholine or verapamil. The addition of vitamin C during hyperoxia augmented FBF responses to acetylcholine. These results suggest that hyperoxic vasoconstriction is mediated by oxidative stress. Moreover, hyperoxia impairs acetylcholine-mediated vasodilation in the setting of intact endothelial function. These effects of hyperoxia are prevented by vitamin C, providing evidence that hyperoxia-derived free radicals impair the activity of endothelium-derived vasoactive factors.     

Towards the co‐ordination of terrestrial ecosystem protocols across European research infrastructures

The study of ecosystem processes over multiple scales of space and time is often best achieved using comparable data from multiple sites. Yet, long‐term ecological observatories have often developed their own data collection protocols. Here, we address this problem by proposing a set of ecological protocols suitable for widespread adoption by the ecological community. Scientists from the European ecological research community prioritized terrestrial ecosystem parameters that could benefit from a more consistent approach to data collection within the resources available at most long‐term ecological observatories. Parameters for which standard methods are in widespread use, or for which methods are evolving rapidly, were not selected. Protocols were developed by domain experts, building on existing methods where possible, and refined through a process of field testing and training. They address above‐ground plant biomass; decomposition; land use and management; leaf area index; soil mesofaunal diversity; soil C and N stocks, and greenhouse gas emissions from soils. These complement existing methods to provide a complete assessment of ecological integrity. These protocols offer integrated approaches to ecological data collection that are low cost and are starting to be used across the European Long Term Ecological Research community.     

Circadian and homeostatic regulation of structural synaptic plasticity in hypocretin neurons

Neurons exhibit rhythmic activity that ultimately affects behavior such as sleep. In living zebrafish larvae, we used time-lapse two-photon imaging of the presynaptic marker synaptophysin in hypocretin/orexin (HCRT) neurons to determine the dynamics of synaptic modifications during the day and night. We observed circadian rhythmicity in synapse number in HCRT axons. This rhythm is regulated primarily by the circadian clock but is also affected by sleep deprivation. Furthermore, NPTX2, a protein implicated in AMPA receptor clustering, modulates circadian synaptic changes. In zebrafish, nptx2b is a rhythmic gene that is mostly expressed in hypothalamic and pineal gland cells. Arrhythmic transgenic nptx2b overexpression (hcrt:NPTX2b) increases synapse number and abolishes rhythmicity in HCRT axons. Finally, hcrt:NPTX2b fish are resistant to the sleep-promoting effects of melatonin. This behavioral effect is consistent with NPTX2b-mediated increased activity of HCRT circuitry. These data provide real-time in vivo evidence of circadian and homeostatic regulation of structural synaptic plasticity.     

Box C/D RNA-guided 2'-O methylations and the intron of tRNATrp are not essential for the viability of Haloferax volcanii

Deleting the box C/D RNA-containing intron in the Haloferax volcanii tRNA^Trp gene abolishes RNA-guided 2′-O methylations of C34 and U39 residues of tRNA^Trp. However, this deletion does not affect growth under standard conditions.     

Biodegradation and detoxification of phenolic compounds by pure and mixed indigenous cultures in aerobic reactors

Degradation and detoxification of a mixture of persistent compounds (2-chlorophenol, phenol and m-cresol) were studied by using pure and mixed indigenous cultures in aerobic reactors. Biodegradation assays were performed in batch and continuous flow reactors. Biodegradation was evaluated by determining total phenols, ultraviolet spectrophotometry and chemical oxygen demand (COD). Microbial growth was measured by the plate count method. Scanning electronic microscopy was employed to observe the microbial community in the reactor. Detoxification was evaluated by using Daphnia magna toxicity tests. Individual compounds were degraded by pure bacteria cultures within 27 h. The mixture of 2-clorophenol (100 mgl⁻¹), phenol (50 mgl⁻¹) and m-cresol (50 mgl⁻¹) was degraded by mixed bacteria cultures under batch conditions within 36 h: 99.8% of total phenols and 92.5% of COD were removed; under continuous flow conditions 99.8% of total phenols and 94.9% of COD were removed. Mineralization of phenolic compounds was assessed by gas chromatography performed at the end of the batch assays and in the effluent of the continuous-flow reactor. Toxicity was not detected in the effluent of the continuous-flow reactor.     

Microbial degradation and detoxification of 2,4-dinitrophenol in aerobic and anoxic processes

A bacterial strain was isolated from a river sediment in Buenos Aires, Argentina, owing to its ability to utilize 2,4-dinitrophenol (2,4-DNP) as the sole carbon, nitrogen and energy source. The strain was identified as Rhodococcus opacus on the basis of its 16S rRNA gene sequence. R. opacus degrades aerobically 0.27 and 0.54 mM within 22 and 28 h, respectively, and releases the nitro groups from 2,4-DNP as nitrites. Aerobic biodegradation processes were performed using a 2-l volume microfermentor at with agitation (200 rpm), and were evaluated by spectrophotometry, high performance liquid chromatography (HPLC) and microbial growth. The absence of 2,4-DNP transformation products was also confirmed by gas chromatography mass spectrometry (GC–MS). As the nitrite released during 2,4-DNP degradation is in addition an environmental toxic agent it was removed by denitrification in an anoxic process. Detoxification was assessed by using luminescent bacteria, algae and seeds toxicity tests. Toxicity was not detected after combining both the aerobic and anoxic processes.     

Aerobic degradation of 3-chlorobenzoic acid by an indigenous strain isolated from a polluted river

An indigenous strain of Pseudomonas putida capable of degrading 3-chlorobenzoic acid as the sole carbon source was isolated from the Riachuelo, a polluted river in Buenos Aires. Aerobic biodegradation assays were performed using a 2-l microfermentor. Biodegradation was evaluated by spectrophotometry, chloride release, gas chromatography and microbial growth. Detoxification was evaluated by using Vibrio fischeri, Pseudokirchneriella subcapitata and Lactuca sativa as test organisms. The indigenous bacterial strain degrades 100 mg l⁻¹ 3-chlorobenzoic acid in 14 h with a removal efficiency of 92.0 and 86.1% expressed as compound and chemical oxygen demand removal, respectively. The strain was capable of degrading up to 1,000 mg of the compound l⁻¹. Toxicity was not detected at the end of the biodegradation process. Besides initial concentration, the effect of different factors, such as initial pH, initial inoculum, adaptation to the compound and presence of other substrates and toxic related compounds, was studied.