Depigmenting effect of Xanthohumol from hop extract in MNT-1 human melanoma cells and normal human melanocytes

Xanthohumol (XH) is the most abundant prenylated flavonoid found in the hop plant (Humulus lupulus L.) and has previously been shown to have depigmenting effects in B16F10 mouse melanoma cells; however, studies of its depigmenting efficacy in human melanocytes are still lacking. In this work, we explored the effects of XH on melanogenesis in MNT-1 human melanoma cells and normal human melanocytes from darkly-pigmented skin (HEM-DP). XH was screened for cytotoxicity over 48 h, and subsequently tested on melanogenesis in MNT-1 cells. XH was further tested in HEM-DP cells for melanin synthesis and melanosome export; dendricity was quantitated to assess effects on melanosome export. Melanosome degradation was studied in human keratinocytes (HaCaT). Our results showed that XH inhibited melanin synthesis in MNT-1 cells at 30 μM but increased intracellular tyrosinase activity without affecting ROS levels. In HEM-DP cells, XH robustly suppressed cellular tyrosinase activity at nontoxic concentrations (2.5–5 μM) without any effect on melanin synthesis. However, XH inhibited melanosome export by reducing dendrite number and total dendrite length. Further testing in HaCaT cells demonstrated that XH induced melanosome degradation at low micromolar concentrations without any cytotoxicity. In summary, our results demonstrate that XH at low micromolar concentrations might hold promise as a potent inhibitor of human pigmentation by primarily targeting melanin export and melanin degradation. Further studies to elucidate the signaling mechanisms of action of melanosome export inhibition by XH and in vivo efficacy are warranted.     

Dispersal and competitive impacts of a third fire ant decapitating fly (Pseudacteon obtusus) established in North Central Florida

Self-sustaining classical biological control agents offer hope for permanent wide-area control of imported Solenopsis fire ants in the United States because escape from abundant natural enemies left behind in Argentina is a likely reason for unusually high fire ant densities in the United States. The fire ant decapitating fly Pseudacteon obtusus Borgmeier (Diptera: Phoridae) was released as a biocontrol agent of the red imported fire ant (Solenopsis invicta Buren) in Gainesville, FL because it is a common parasitoid of this ant in Argentina and because it has a higher propensity of attacking fire ants along foraging trails than the two Pseudacteon species previously released. Field surveys of a rapidly expanding P. obtusus population (8–12 km/yr) proved that this fly was capable of thriving and successfully competing with the much more abundant Pseudacteon curvatus Borgmeier. However, Pseudacteon tricuspis Borgmeier, the first decapitating fly released, was effectively excluded from most sample sites when faced with competition from both P. curvatus and the similar-sized P. obtusus. Despite clear evidence for competitive exclusion, P. tricuspis abundance at sample sites was positively correlated with the abundance of its two competitors—probably because of moderate to strong covariability in the suitability of sample sites for all three congeners. The addition of P. curvatus, the second parasitoid released, increased total parasitism pressure on fire ant populations by about 10-fold. The addition of P. obtusus, the third species, did not measurably improve total guild parasitism rates on imported fire ants in North Central Florida (as assessed by roadside trap counts), but the performance of this species will likely vary with habitat, region, and climate.     

Huntington's Disease Induced Cardiac Amyloidosis Is Reversed by Modulating Protein Folding and Oxidative Stress Pathways in the Drosophila Heart

Amyloid-like inclusions have been associated with Huntington''s disease (HD), which is caused by expanded polyglutamine repeats in the Huntingtin protein. HD patients exhibit a high incidence of cardiovascular events, presumably as a result of accumulation of toxic amyloid-like inclusions. We have generated a Drosophila model of cardiac amyloidosis that exhibits accumulation of PolyQ aggregates and oxidative stress in myocardial cells, upon heart-specific expression of Huntingtin protein fragments (Htt-PolyQ) with disease-causing poly-glutamine repeats (PolyQ-46, PolyQ-72, and PolyQ-102). Cardiac expression of GFP-tagged Htt-PolyQs resulted in PolyQ length-dependent functional defects that included increased incidence of arrhythmias and extreme cardiac dilation, accompanied by a significant decrease in contractility. Structural and ultrastructural analysis of the myocardial cells revealed reduced myofibrillar content, myofibrillar disorganization, mitochondrial defects and the presence of PolyQ-GFP positive aggregates. Cardiac-specific expression of disease causing Poly-Q also shortens lifespan of flies dramatically. To further confirm the involvement of oxidative stress or protein unfolding and to understand the mechanism of PolyQ induced cardiomyopathy, we co-expressed expanded PolyQ-72 with the antioxidant superoxide dismutase (SOD) or the myosin chaperone UNC-45. Co-expression of SOD suppressed PolyQ-72 induced mitochondrial defects and partially suppressed aggregation as well as myofibrillar disorganization. However, co-expression of UNC-45 dramatically suppressed PolyQ-72 induced aggregation and partially suppressed myofibrillar disorganization. Moreover, co-expression of both UNC-45 and SOD more efficiently suppressed GFP-positive aggregates, myofibrillar disorganization and physiological cardiac defects induced by PolyQ-72 than did either treatment alone. Our results demonstrate that mutant-PolyQ induces aggregates, disrupts the sarcomeric organization of contractile proteins, leads to mitochondrial dysfunction and increases oxidative stress in cardiomyocytes leading to abnormal cardiac function. We conclude that modulation of both protein unfolding and oxidative stress pathways in the Drosophila heart model can ameliorate the detrimental PolyQ effects, thus providing unique insights into the genetic mechanisms underlying amyloid-induced cardiac failure in HD patients.     

Ferristatin II Promotes Degradation of Transferrin Receptor-1 In Vitro and In Vivo

Previous studies have shown that the small molecule iron transport inhibitor ferristatin (NSC30611) acts by down-regulating transferrin receptor-1 (TfR1) via receptor degradation. In this investigation, we show that another small molecule, ferristatin II (NSC8679), acts in a similar manner to degrade the receptor through a nystatin-sensitive lipid raft pathway. Structural domains of the receptor necessary for interactions with the clathrin pathway do not appear to be necessary for ferristatin II induced degradation of TfR1. While TfR1 constitutively traffics through clathrin-mediated endocytosis, with or without ligand, the presence of Tf blocked ferristatin II induced degradation of TfR1. This effect of Tf was lost in a ligand binding receptor mutant G647A TfR1, suggesting that Tf binding to its receptor interferes with the drug’s activity. Rats treated with ferristatin II have lower TfR1 in liver. These effects are associated with reduced intestinal ⁵⁹Fe uptake, lower serum iron and transferrin saturation, but no change in liver non-heme iron stores. The observed hypoferremia promoted by degradation of TfR1 by ferristatin II appears to be due to induced hepcidin gene expression.     

Investment risk in bioenergy crops

Perennial, cellulosic bioenergy crops represent a risky investment. The potential for adoption of these crops depends not only on mean net returns, but also on the associated probability distributions and on the risk preferences of farmers. Using 6‐year observed crop yield data from highly productive and marginally productive sites in the southern Great Lakes region and assuming risk neutrality, we calculate expected breakeven biomass yields and prices compared to corn (Zea mays L.) as a benchmark. Next we develop Monte Carlo budget simulations based on stochastic crop prices and yields. The crop yield simulations decompose yield risk into three components: crop establishment survival, time to maturity, and mature yield variability. Results reveal that corn with harvest of grain and 38% of stover (as cellulosic bioenergy feedstock) is both the most profitable and the least risky investment option. It dominates all perennial systems considered across a wide range of farmer risk preferences. Although not currently attractive for profit‐oriented farmers who are risk neutral or risk averse, perennial bioenergy crops have a higher potential to successfully compete with corn under marginal crop production conditions.     

MiR-449a Affects Epithelial Proliferation during the Pseudoglandular and Canalicular Phases of Avian and Mammal Lung Development

Congenital diaphragmatic hernia is associated with pulmonary hypoplasia and respiratory distress, which result in high mortality and morbidity. Although several transgenic mouse models of lung hypoplasia exist, the role of miRNAs in this phenotype is incompletely characterized. In this study, we assessed microRNA expression levels during the pseudoglandular to canalicular phase transition of normal human fetal lung development. At this critical time, when the distal respiratory portion of the airways begins to form, microarray analysis showed that the most significantly differentially expressed miRNA was miR-449a. Prediction algorithms determined that N-myc is a target of miR-449a and identified the likely miR-449a:N-myc binding sites, confirmed by luciferase assays and targeted mutagenesis. Functional ex vivo knock-down in organ cultures of murine embryonic lungs, as well as in ovo overexpression in avian embryonic lungs, suggested a role for miR-449a in distal epithelial proliferation. Finally, miR-449a expression was found to be abnormal in rare pulmonary specimens of human fetuses with Congenital Diaphragmatic Hernia in the pseudoglandular or canalicular phase. This study confirms the conserved role of miR-449a for proper pulmonary organogenesis, supporting the delicate balance between expansion of progenitor cells and their terminal differentiation, and proposes the potential involvement of this miRNA in human pulmonary hypoplasia.     

Prolonged Cerebral Circulation Time Is the Best Parameter for Predicting Vasospasm during Initial CT Perfusion in Subarachnoid Hemorrhagic Patients

Purpose

We sought to imitate angiographic cerebral circulation time (CCT) and create a similar index from baseline CT perfusion (CTP) to better predict vasospasm in patients with subarachnoid hemorrhage (SAH).

Methods

Forty-one SAH patients with available DSA and CTP were retrospectively included. The vasospasm group was comprised of patients with deterioration in conscious functioning and newly developed luminal narrowing; remaining cases were classified as the control group. The angiography CCT (XA-CCT) was defined as the difference in TTP (time to peak) between the selected arterial ROIs and the superior sagittal sinus (SSS). Four arterial ROIs were selected to generate four corresponding XA-CCTs: the right and left anterior cerebral arteries (XA-CCT_RA2 and XA-CCT_LA2) and right- and left-middle cerebral arteries (XA-CCT_RM2 and XA-CCT_LM2). The CCTs from CTP (CT-CCT) were defined as the differences in TTP from the corresponding arterial ROIs and the SSS. Correlations of the different CCTs were calculated and diagnostic accuracy in predicting vasospasm was evaluated.

Results

Intra-class correlations ranged from 0.96 to 0.98. The correlations of XA-CCT_RA2, XA-CCT_RM2, XA-CCT_LA2, and XA-CCT_LM2 with the corresponding CT-CCTs were 0.64, 0.65, 0.53, and 0.68, respectively. All CCTs were significantly prolonged in the vasospasm group (5.8–6.4 s) except for XA-CCT_LA2. CT-CCT_A2 of 5.62 was the optimal cut-off value for detecting vasospasm with a sensitivity of 84.2% and specificity 82.4%

Conclusion

CT-CCTs can be used to interpret cerebral flow without deconvolution algorithms, and outperform both MTT and TTP in predicting vasospasm risk. This finding may help facilitate management of patients with SAH.     

Evolution and ecology of feeding in elasmobranchs

Paleozoic chondrichthyans had a large gape, numerous spike-liketeeth, limited cranial kinesis, and a non-suspensory hyoid,suggesting a feeding mechanism dominated by bite and ram. Modernsharks are characterized by a mobile upper jaw braced by a suspensoryhyoid arch that is highly kinetic. In batoids, the upper jawis dissociated from the cranium permitting extensive protrusionof the jaws. Similar to actinopterygians, the evolution of highlymobile mandibular and hyoid elements has been correlated withextensive radiation of feeding modes in elasmobranchs, particularlythat of suction. Modern elasmobranchs possess a remarkable varietyof feeding modes for a group containing so few species. Biting,suction or filter-feeding may be used in conjunction with ramto capture prey, with most species able to use a combinationof behaviors during a strike. Suction-feeding has repeatedlyarisen within all recent major elasmobranch clades and is associatedwith a suite of morphological and behavioral specializations.Prey capture in a diverse assemblage of purported suction-feedingelasmobranchs is investigated in this study. Drop in water pressuremeasured in the mouth and at the location of the prey showsthat suction inflow drops off rapidly with distance from thepredator's mouth. Elasmobranchs specializing in suction-feedingmay be limited to bottom associated prey and because of theirsmall gape may have a diet restricted to relatively small prey.Behavior can affect performance and overcome constraints imposedby the fluid medium. Suction performance can be enhanced byproximity to a substrate or by decreasing distance from predatorto prey using various morphological and/or behavioral characteristics.Benthic suction-feeders benefit by the increased strike radiusdue to deflection of water flow when feeding close to a substrate,and perhaps require less accuracy when capturing prey. Suctionand ram-suction-feeding elasmobranchs can also use suction inflowto draw prey to them from a short distance, while ram-feedingsharks must accelerate and overtake the prey. The relationshipbetween feeding strategy and ecology may depend in part on ecological,mechanistic or evolutionary specialization. Mechanistic suction-feedingspecialist elasmobranchs are primarily benthic, while most epibenthicand pelagic elasmobranchs are generalists and use ram, suction,and biting to catch a diversity of prey in various habitats.Some shark species are considered to be ecological specialistsin choosing certain kinds of prey over others. Batoids are evolutionaryspecialists in having a flattened morphology and most are generalistfeeders. Filter-feeding elasmobranchs are ecological, mechanistic,and evolutionary specialists.