Sequence and functional characterization of hypoxia inducible factors, HIF1α, HIF2αa, and HIF3α, from the estuarine fish, Fundulus heteroclitus

Author Posting. © The Author(s), 2016. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in American Journal of Physiology-Regulatory Integrative and Comparative Physiology, 312 (2017): R412-R425, doi:10.1152/ajpregu.00402.2016.The hypoxia inducible factor (HIF) family of transcription factors plays central roles in the development, physiology, pathology, and environmental adaptation of animals. Because many aquatic habitats are characterized by episodes of low dissolved oxygen, fish represent ideal models to study the roles of HIF in the response to aquatic hypoxia. The estuarine fish Fundulus heteroclitus occurs in habitats prone to hypoxia, it responds to low oxygen via behavioral, physiological, and molecular changes, and one member of the HIF family, HIF2α, has been previously described. Herein, cDNA sequencing, phylogenetic analyses, and genomic approaches were used to determine other members of the HIFα family from F. heteroclitus and their relationships to HIFα subunits from other vertebrates. In vitro and cellular approaches demonstrated that full-length forms of HIF1α, 2α, and 3α independently formed complexes with the β subunit (ARNT) to bind to hypoxia response elements and activate reporter gene expression. Quantitative PCR showed that HIFα mRNA abundance varied among organs of normoxic fish in an isoform-specific fashion. Analysis of the F. heteroclitus genome revealed a locus encoding a second HIF2α, HIF2αb, a predicted protein lacking oxygen sensing and transactivation domains. Finally, sequence analyses demonstrated polymorphism in the coding sequence of each F. heteroclitus HIFα subunit, suggesting that genetic variation in these transcription factors may play a role in the variation in hypoxia responses among individuals or populations.This research was supported in part by the National Science Foundation (IBN-0236494 and DEB-1120263) and by National Institute of Environmental Health Sciences (NIEHS) grant P42ES007381 (Superfund Basic Research Program at Boston University). Data interpretation was aided by reference to a preliminary draft of the F. heteroclitus genome sequence, which was supported by funding from the National Science Foundation (collaborative research grants DEB-1120512, DEB-1265282, DEB-1120013, DEB-1120263, DEB-1120333, DEB-1120398)

Age-related peculiarities of changes in functional state of a person in the recreation areas

The article is devoted to the study of physiological effects of staying in the park areas in people of different ages. The study involved 20 students aged 18-20 years; 20 schoolchildren aged 14-15 years; 11 elderly people aged 69-76 years. We recorded tidal volume and respiratory rate, took blood pressure and counted heart rate, calculated hemodynamic parameters. We analyzed the parameters of attention, determined the capacity of short-term auditory and visual memory. All studied parameters were recorded before and after a walk in the park. While people spend time in the open air in recreation areas, the positive dynamics in cardiorespiratory system is observed: a decrease in blood pressure and heart rate, but maximal pulmonary ventilation increases. Also, we noticed that the level of state anxiety has reduced. We have discovered that the influence of recreation area environment on the functional state of test subjects depends on their age The magnitude of the changes depends on the deviation of the initial parameters from the normal values. The most positive values are observed in people with maximum deviation from the norm. The practical significance of the study is to expand the range of drug-free methods to enhance functions of cardiovascular, respiratory systems and the brain in elderly people with the help of sensory environment of recreation areas

Human resource development in the implementation of the concept of “smart cities”

The improvement of urban systems based on the active use of information technologies, which is necessary to achieve these goals, requires specialists with appropriate qualifications. This article uses a methodological approach to the study of human resource development from the perspective of two-way communication between producers and consumers of an innovative product. The educational, structural-industry and information technology aspects of development are considered. The results of the analysis are presented, and the methods of partnership cooperation between higher educational institutions and representatives of the business community in the construction and housing and utilities sectors are identified. The experience in the field of training and retraining specialists in construction specialties with elements of student research has been synthesized. The prospects for improving the system of personnel training in the field of housing and communal services are identified. A SWOT analysis of the strengths and weaknesses of urban modernization has been carried out. Among the weaknesses, the failure of highly qualified specialists was revealed. Among the threats is the absence of a legislative and methodological framework in many areas of housing and communal services reforming. In the annex to the concept of “smart cities”, Rohmer’s model is considered, confirming the primary role of human resources in reforming and further operating the urban infrastructure. It is shown that the success of the initiative of a smart city depends on personnel, processes and technologies

Population proteomics: quantitative variation within and among populations in cardiac protein expression

Population analysis of gene expression is typically achieved by quantifying levels of mRNA; however, gene expression is also a function of protein translation and turnover. Therefore, a complete understanding of population variation in gene expression requires quantitative knowledge of protein expression within and among natural populations. We used two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) to quantitatively compare expression of heart ventricle proteins among 18 individuals in three populations of the teleost fish Fundulus. Among populations, expressions between orthologous proteins and mRNAs were generally positively correlated. Additionally, similar to the pattern of cardiac mRNA expression for the same populations, we found considerable variation in protein expression both within and among populations: Of 408 protein features in 2D gels, 34% are significantly different (P < 0.01) among individuals within a population, 9% differ between populations, and 12% have a pattern of expression that suggests they have evolved by natural selection. Although similar to mRNA expression, the frequency of significant differences among populations is larger for proteins. Similar to mRNA expressions, expressions of most proteins are correlated to the expressions of many other proteins. However, the correlations among proteins are more extensive than the correlation for similar RNAs. These correlations suggest a greater coordinate regulation of protein than mRNA expression. The larger frequency of significant differences among populations and the greater frequency of correlated expression among proteins versus among RNAs suggest that the molecular mechanisms affecting protein expression enhance the differences among populations, and these regulatory steps could be a source of variation for adaptation

Nutritional Contingency Reduces Alcohol Drinking by Altering Central Neurotransmitter Receptor Gene Expression in Rats

We have previously shown that 6 weeks of intermittent high-fat diet (Int-HFD) pre-exposure significantly reduced alcohol drinking in rats, providing preliminary evidence of the effectiveness of a dietary intervention in reducing alcohol intake. However, the functional framework and underlying neurobiological mechanisms of such dietary intervention are unknown. Here, we examined the impact of Int-HFD pre-exposure duration on alcohol drinking, plasma feeding peptides, and central neurotransmitter receptors gene expression. Male Long Evans rats (n = 6&ndash;7/group) received no pre-exposure, 1 or 2 weeks pre-exposure to Int-HFD and alcohol drinking (two-bottle choice) was evaluated. We observed HFD pre-exposure-dependent decrease in alcohol drinking, with a significant decrease observed following 2 weeks of Int-HFD pre-exposure. No significant between-group differences in plasma feeding peptides (i.e., ghrelin, leptin, insulin) were detected. A PCR array revealed that the expression of several neurotransmitter receptors was significantly (p &lt; 0.05 and &ge;2-fold) altered in the striatum and ventral tegmental area compared to controls. These data suggest that pre-exposure to a palatable diet is critical to reduce alcohol drinking in rats, possibly through genetic alterations in the brain reward circuitry. Importantly, the present study is a step forward in identifying the critical framework needed to evaluate the therapeutic potential of nutritional contingency in the management of alcoholism

Endogenous expression of inactive lysine deacetylases reveals deacetylation-dependent cellular mechanisms.

Acetylation of lysine residues is an important and common post-translational regulatory mechanism occurring on thousands of non-histone proteins. Lysine deacetylases (KDACs or HDACs) are a family of enzymes responsible for removing acetylation. To identify the biological mechanisms regulated by individual KDACs, we created HT1080 cell lines containing chromosomal point mutations, which endogenously express either KDAC6 or KDAC8 having single inactivated catalytic domain. Engineered HT1080 cells expressing inactive KDA6 or KDAC8 domains remained viable and exhibited enhanced acetylation on known substrate proteins. RNA-seq analysis revealed that many changes in gene expression were observed when KDACs were inactivated, and that these gene sets differed significantly from knockdown and knockout cell lines. Using GO ontology, we identified several critical biological processes associated specifically with catalytic activity and others attributable to non-catalytic interactions. Treatment of wild-type cells with KDAC-specific inhibitors Tubastatin A and PCI-34051 resulted in gene expression changes distinct from those of the engineered cell lines, validating this approach as a tool for evaluating in-cell inhibitor specificity and identifying off-target effects of KDAC inhibitors. Probing the functions of specific KDAC domains using these cell lines is not equivalent to doing so using previously existing methods and provides novel insight into the catalytic functions of individual KDACs by investigating the molecular and cellular changes upon genetic inactivation

Glyceollin Effects on MRP2 and BCRP in Caco-2 Cells, and Implications for Metabolic and Transport Interactions

Glyceollins are phytoalexins produced in soybeans under stressful growth conditions. On the basis of prior evaluations, they show potential to treat multiple diseases, including certain cancers, Type 2 diabetes, and cardiovascular conditions. The aim of the present study was to expand on recent studies designed to initially characterize the intestinal disposition of glyceollins. Specifically, studies were undertaken in Caco-2 cells to evaluate glyceollins\u27 effects on apical efflux transporters, namely, MRP2 and BCRP, which are the locus of several intestinal drug-drug and drug-food interactions. 5- (and 6)-carboxy-2\u27,7\u27-dichloroflourescein (CDF) was used to provide a readout on MRP2 activity, whereas BODIPY-prazosin provided an indication of BCRP alteration. Glyceollins were shown to reverse MRP2-mediated CDF transport asymmetry in a concentration-dependent manner, with activity similar to the MRP2 inhibitor, MK-571. Likewise, they demonstrated concentration-dependent inhibition of BCRP-mediated efflux of BODIPY-prazosin with a potency similar to that of Ko143. Glyceollin did not appreciably alter MRP2 or BCRP expression following 24 h of continuous exposure. The possibility that glyceollin mediated inhibition of genistein metabolite efflux by either transporter was evaluated. However, results demonstrated an interaction at the level of glyceollin inhibition of genistein metabolism rather than inhibition of metabolite transport

Glyceollin Effects on MRP2 and BCRP in Caco-2 Cells, and Implications for Metabolic and Transport Interactions

The study examined the impact of visual predictability on dual-task performance in driving and tracking tasks. Participants (N = 27) performed a simulated driving task and a pursuit tracking task. In either task, visual predictability was manipulated by systematically varying the amount of advance visual information: in the driving task, participants drove at night with low beam, at night with high beam, or in daylight; in the tracking task, participants saw a white line that specified the future target trajectory for 200, 400 or 800 ms. Concurrently with driving or tracking, participants performed an auditory task. They had to discriminate between two sounds and press a pedal upon hearing the higher sound. Results show that in general, visual predictability benefited driving and tracking; however, dual-task driving performance was best with highest visual predictability (daylight), dual-task tracking performance was best with medium visual predictability (400 ms). Braking/reaction times were higher in dual tasks compared to single tasks, but were unaffected by visual predictability, showing that its beneficial effects did not transfer to the auditory task. In both tasks, manual accuracy decreased around the moment the foot pressed the pedal, indicating interference between tasks. We, therefore, conclude that despite a general beneficial impact of predictability, the integration of visual information seems to be rather task specific, and that interference between driving and audiomotor tasks, and tracking and audiomotor tasks, seems comparable