As neuronal subtypes are increasingly categorized, delineating their functional role is paramount. The preBötzinger complex (preBötC) subpopulation expressing the neuropeptide somatostatin (SST) is ...classified as mostly excitatory, inspiratory‐modulated and not rhythmogenic. We further characterized their phenotypic identity: 87% were glutamatergic and the balance were glycinergic and/or GABAergic. We then used optogenetics to investigate their modulatory role in both anaesthetized and freely moving mice. In anaesthetized mice, short photostimulation (100 ms) of preBötC SST+ neurons modulated breathing‐related variables in a combinatory phase‐ and state‐dependent manner; changes in inspiratory duration, inspiratory peak amplitude (Amp), and phase were different at higher (≥2.5 Hz) vs. lower (<2.5 Hz) breathing frequency (f). Moreover, we observed a biphasic effect of photostimulation during expiration that is probabilistic, that is photostimulation given at the same phase in consecutive cycles can evoke opposite responses (lengthening vs. shortening of the phase). These unexpected probabilistic state‐ and phase‐dependent responses to photostimulation exposed properties of the preBötC that were not predicted and cannot be readily accounted for in current models of preBötC pattern generation. In freely moving mice, prolonged photostimulation decreased f in normoxia, hypoxia or hypercapnia, and increased Amp and produced a phase advance, which was similar to the results in anaesthetized mice when f ≥ 2.5 Hz. We conclude that preBötC SST+ neurons are a key mediator of the extraordinary and essential lability of breathing pattern.
Key points
PreBötzinger complex (preBötC) SST+ neurons, which modulate respiratory pattern but are not rhythmogenic, were transfected with channelrhodopsin to investigate phase‐ and state‐dependent modulation of breathing pattern in anaesthetized and freely behaving mice in normoxia, hypoxia and hypercapnia.
In anaesthetized mice, photostimulation during inspiration increased inspiratory duration and amplitude regardless of baseline f, yet the effects were more robust at higher f.
In anaesthetized mice with low f (<2.5 Hz), photostimulation during expiration evoked either phase advance or phase delay, whereas in anaesthetized mice with high f (≥2.5 Hz) and in freely behaving mice in normoxia, hypoxia or hypercapnia, photostimulation always evoked phase advance.
Phase‐ and state‐dependency is a function of overall breathing network excitability.
The f‐dependent probabilistic modulation of breathing pattern by preBötC SST+ neurons was unexpected, requiring reconsideration of current models of preBötC function, which neither predict nor can readily account for such responses.
figure legend preBötC SST+ neurons were optogenetically stimulated to investigate phase‐ and state‐dependent modulation of breathing pattern in anaesthetized mice. In the same mouse, the same stimulus evoked either phase advance (shortening of the cycle), or phase delay (prolongation of the cycle) or no change. Using cophase plots (phase φ vs. cophase θ), it was determined that the transition between a bimodal effect (phase advance or delay) and a unimodal effect (phase advance) occurred at baseline breathing f ≈ 2.5 Hz.
► Cheese whey was used as carbon source for Chlorella vulgaris growth. ► Mixotrophic microalgae grew faster than photoautotrophic cells. ► Maximum starch productivity was achieved under mixotrophic ...conditions. ► Highest pigment content (0.74%) was obtained in the photoautotrophic culture.
Growth parameters and biochemical composition of the green microalga Chlorella vulgaris cultivated under different mixotrophic conditions were determined and compared to those obtained from a photoautotrophic control culture. Mixotrophic microalgae showed higher specific growth rate, final biomass concentration and productivities of lipids, starch and proteins than microalgae cultivated under photoautotrophic conditions. Moreover, supplementation of the inorganic culture medium with hydrolyzed cheese whey powder solution led to a significant improvement in microalgal biomass production and carbohydrate utilization when compared with the culture enriched with a mixture of pure glucose and galactose, due to the presence of growth promoting nutrients in cheese whey. Mixotrophic cultivation of C. vulgaris using the main dairy industry by-product could be considered a feasible alternative to reduce the costs of microalgal biomass production, since it does not require the addition of expensive carbohydrates to the culture medium.
Human Papillomavirus (HPV) is the most common sexually transmitted virus. Worldwide, the most common high-risk (HR)-HPV are -16/18, and approximately 70% of cervical cancers (CC) are due to infection ...by these genotypes. Persistent infection by HR-HPV is a necessary but not sufficient cause of this cancer, which develops over a long period through precursor lesions, which can be detected by cytological screening. Although this screening has decreased the incidence of CC, HPV-related cervical disease, including premalignant and malignant lesions, continues to be a major burden on health-care systems. Although not completely elucidated, the HPV-driven molecular mechanisms underlying the development of cervical lesions have provided a number of potential biomarkers for both diagnostic and prognostic use in the clinical management of women with HPV-related cervical disease, and these biomarkers can also be used to increase the positive predictive value of current screening methods. In addition, they can provide insights into the biology of HPV-induced cancer and thus lead to the development of nonsurgical therapies. Considering the importance of detecting HPV and related biomarkers, a variety of methods are being developed for these purposes. This review summarizes current knowledge of detection methods for HPV, and related biomarkers that can be used to discriminate lesions with a high risk of progression to CC.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
This work reports a thorough molecular dynamics investigation on the aggregation patterns of curcumin and piperine in water, ethanol and a mixture of both solvents. The low solubility of curcumin in ...water results in a rapid formation of very stable dimers for both keto and enol tautomers. In agreement with a higher solubility, piperine molecules move closer and farther apart several times during the simulation, which indicates the formation of a less stable dimer in water. In contrast, both curcumin and piperine are soluble in ethanol and, thus, dimers can hardly be formed in this media. In comparison with a pure-water solvent, a 30 : 70 mixture of ethanol and water significantly reduces the probability of formation of most dimers of curcumin and piperine molecules. The simulations show that larger clusters may be complex structures, but the formation of stacks (in the case of piperine and enol tautomer of curcumin) and cages (when the keto tautomer of curcumin is involved) are not rare. Furthermore, it is shown that each single molecule presents a certain degree of mobility in the cluster, especially on the surface, but without leading to dissociation.
Molecular dynamics simulations show that the aggregates resulting from mixtures of curcumin and piperine in water mainly form stacked or cage structures with a great fluxional character, which is enhanced in the presence of ethanol molecules.
Increasing microalgal starch content by nutrient limitation has been regarded as an affordable approach for the production of third generation bioethanol. This work evaluated starch accumulation in
...Chlorella vulgaris P12 under different initial concentrations of nitrogen (0–2.2
g
urea
L
−1) and iron (0–0.08
g
FeNa-EDTA
L
−1) sources, using a central composite design (CCD) for two factors. The obtained model:
Starch content (%)
=
8.220
−
16.133
X
1
+
13.850
X
1
2
, relating starch accumulation in microalgae with the coded level for initial urea concentration in the growth medium (
X
1) presented a good concordance between the predicted and experimental values (
R
2
=
0.94). Since accumulation of starch occurred at nitrogen depletion conditions under which the cell growth was much slower than that observed during nitrogen supplemented cultivations, a two-stage cultivation process for high starch accumulation (>40%) and cell growth of
C. vulgaris was proposed: a first cultivation stage using nitrogen- and iron-supplemented medium (initial urea and FeNa-EDTA concentrations of 1.1 and 0.08
g
L
−1, respectively), followed by a second cultivation stage in a nitrogen- and iron-free medium. The high starch content obtained suggests
C. vulgaris P12 as a very promising feedstock for bioethanol production.
Context: Kisspeptin, encoded by the KISS1 gene, is a key stimulatory factor of GnRH secretion and puberty onset. Inactivating mutations of its receptor (KISS1R) cause isolated hypogonadotropic ...hypogonadism (IHH). A unique KISS1R-activating mutation was described in central precocious puberty (CPP).
Objective: Our objective was to investigate KISS1 mutations in patients with idiopathic CPP and normosmic IHH.
Patients: Eighty-three children with CPP (77 girls) and 61 patients with IHH (40 men) were studied. The control group consisted of 200 individuals with normal pubertal development.
Methods: The promoter region and the three exons of KISS1 were amplified and sequenced. Cells expressing KISS1R were stimulated with synthetic human wild-type or mutant kisspeptin-54 (kp54), and inositol phosphate accumulation was measured. In a second set of experiments, kp54 was preincubated in human serum before stimulation of the cells.
Results: Two novel KISS1 missense mutations, p.P74S and p.H90D, were identified in three unrelated children with idiopathic CPP. Both mutations were absent in 400 control alleles. The p.P74S mutation was identified in the heterozygous state in a boy who developed CPP at 1 yr of age. The p.H90D mutation was identified in the homozygous state in two unrelated girls with CPP. In vitro studies revealed that the capacity of the P74S and H90D mutants to stimulate IP production was similar to the wild type. After preincubation of wild-type and mutant kp54 in human serum, the capacity to stimulate signal transduction was significantly greater for P74S compared with the wild type, suggesting that the p.P74S variant is more stable. Only polymorphisms were found in the IHH group.
Conclusion: Two KISS1 mutations were identified in unrelated patients with idiopathic CPP. The p.P74S variant was associated with higher kisspeptin resistance to degradation in comparison with the wild type, suggesting a role for this mutation in the precocious puberty phenotype.
A mutation in the KISS1 gene (P74S), identified in a boy with central precocious puberty, results in higher kisspeptin resistance to degradation in vitro.
Aim
Future climate changes may affect species distribution and their genetic diversity, hampering species adaptation to a new climate or tracking the suitable conditions. Amphibians have high ...sensitivity to environmental degradation and changes in temperature and humidity. Thus, the expected climatic changes by the end‐of‐century (EOC 2100) may cause local or complete extinction of some species. Here, we address the effects of climate change on genetic and phylogeographical diversity, together with the geographical distribution of the South American treefrog Scinax squalirostris Lutz, 1925. Furthermore, we assess how protected areas will conserve its genetic variation.
Location
South America.
Methods
We combined Ecological Niche Modelling and genetic simulations to predict the effects of climate change on the geographical distribution, genetic diversity, structure and phylogeographical diversity of Scinax squalirostris, using two scenarios of CO2 emission. We also performed a spatial analysis to investigate the effectiveness of the current Protected Areas (PAs) to preserve the species’ genetic and phylogeographical diversity.
Results
Scinax squalirostris' geographical range will potentially increase in the future due to the expansion of suitable areas towards its southern distribution, despite the shrinking of suitable areas in the northern part of its current distribution. Besides the shifts in suitable areas, our findings point to a genetic homogenization across the geographical range of S. squalirostris due to the displacement and loss of genetic ancestry clusters. Although existing PAs are conserving the current genetic diversity, they conserve less phylogeographical diversity than expected by chance. Scinax squalirostris may shift its distribution into areas with lower number of PAs, compromising its future conservation.
Main conclusions
Climate change will potentially increase S. squalirostris range size, however, not towards regions where most of the current established PAs are located, hence driving to homogenization and loss of genetic diversity, and leading to a gap of conservation within PAs.
Mkrn3, the maternally imprinted gene encoding the makorin RING-finger protein-3, has recently emerged as putative pubertal repressor, as evidenced by central precocity caused by MKRN3 mutations in ...humans; yet, the molecular underpinnings of this key regulatory action remain largely unexplored. We report herein that the microRNA, miR-30, with three binding sites in a highly conserved region of its 3' UTR, operates as repressor of Mkrn3 to control pubertal onset. Hypothalamic miR-30b expression increased, while Mkrn3 mRNA and protein content decreased, during rat postnatal maturation. Neonatal estrogen exposure, causing pubertal alterations, enhanced hypothalamic Mkrn3 and suppressed miR-30b expression in female rats. Functional in vitro analyses demonstrated a strong repressive action of miR-30b on Mkrn3 3' UTR. Moreover, central infusion during the juvenile period of target site blockers, tailored to prevent miR-30 binding to Mkrn3 3' UTR, reversed the prepubertal down-regulation of hypothalamic Mkrn3 protein and delayed female puberty. Collectively, our data unveil a novel hypothalamic miRNA pathway, involving miR-30, with a prominent role in the control of puberty via Mkrn3 repression. These findings expand our current understanding of the molecular basis of puberty and its disease states.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
•Parameterizing the canopy cover curve supports accurate ET and yield estimations.•Accuracy of predictions improves when parameterization includes soil water or ET observations.•Good biomass and ...yield predictions were obtained after careful model parameterization.•When using model default parameters predictions are less accurate but acceptable.•The model is good for biomass and yield predictions but less good for soil water simulations.
Several maize field experiments, including deficit and full irrigation, were performed in Ribatejo region, Portugal and were used to assess water stress impacts on yields using the AquaCrop model. The model was assessed after its parameterization using field observations relative to leaf area index (LAI), crop evapotranspiration, soil water content, biomass and final yield data and also using default parameters. LAI data were used to calibrate the canopy cover (CC) curve. Results showed that when the CC curve is properly calibrated, with root mean square errors (RMSE) smaller than 7.4%, model simulations, namely relative to crop evapotranspiration and its partition, show an improved accuracy. The model performance relative to soil water balance simulation revealed a bias in estimation but low estimation errors, with RMSE<13% of the total available soil water. However the model tends to overestimate transpiration and underestimate soil evaporation. A good model performance was obtained relative to biomass and yield predictions, with RMSE lower than 11% and 9% of the average observed biomass and yield, respectively. Overall results show adequacy of AquaCrop for estimating maize biomass and yield under deficit irrigation conditions, mainly when an appropriate parameterization is adopted. The model showed less good performance when using the default parameters but errors are likely acceptable when field data are not available.
The Auger Engineering Radio Array (AERA) is part of the Pierre Auger Observatory and is used to detect the radio emission of cosmic-ray air showers. These observations are compared to the data of the ...surface detector stations of the Observatory, which provide well-calibrated information on the cosmic-ray energies and arrival directions. The response of the radio stations in the 30-80 MHz regime has been thoroughly calibrated to enable the reconstruction of the incoming electric field. For the latter, the energy deposit per area is determined from the radio pulses at each observer position and is interpolated using a two-dimensional function that takes into account signal asymmetries due to interference between the geomagnetic and charge-excess emission components. The spatial integral over the signal distribution gives a direct measurement of the energy transferred from the primary cosmic ray into radio emission in the AERA frequency range. We measure 15.8 MeV of radiation energy for a 1 EeV air shower arriving perpendicularly to the geomagnetic field. This radiation energy-corrected for geometrical effects-is used as a cosmic-ray energy estimator. Performing an absolute energy calibration against the surface-detector information, we observe that this radio-energy estimator scales quadratically with the cosmic-ray energy as expected for coherent emission. We find an energy resolution of the radio reconstruction of 22% for the data set and 17% for a high-quality subset containing only events with at least five radio stations with signal.
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