Miracle-dz/UniFAR-FTUs huggingface dataset:
Phosphorylation of Human Cytochrome P450c17 by p38α Selectively Increases 17,20 Lyase Activity and Androgen Biosynthesis*

Background: Ser/Thr phosphorylation of P450c17 increases 17,20 lyase activity and androgenic capacity. Results: Drug inhibition and siRNA knockdowns in adrenal cells implicate p38α, which phosphorylated bacterially expressed P450c17, doubling 17,20 lyase activity. Conclusion: Phosphorylation of P450c17 by p38α provides a post-translational mechanism distinguishing glucocorticoid from sex steroid synthesis. Significance: p38α pathways may participate in hyperandrogenic states and provide targets for glucocorticoid-sparing inhibition of androgen synthesis. Cytochrome P450c17, a steroidogenic enzyme encoded by the CYP17A1 gene, catalyzes the steroid 17α-hydroxylation needed for glucocorticoid synthesis, which may or may not be followed by 17,20 lyase activity needed for sex steroid synthesis. Whether or not P450c17 catalyzes 17,20 lyase activity is determined by three post-translational mechanisms influencing availability of reducing equivalents donated by P450 oxidoreductase (POR). These are increased amounts of POR, the allosteric action of cytochrome b5 to promote POR-P450c17 interaction, and Ser/Thr phosphorylation of P450c17, which also appears to promote POR-P450c17 interaction. The kinase(s) that phosphorylates P450c17 is unknown. In a series of kinase inhibition experiments, the pyridinyl imidazole drugs SB202190 and SB203580 inhibited 17,20 lyase but not 17α-hydroxylase activity in human adrenocortical HCI-H295A cells, suggesting an action on p38α or p38β. Co-transfection of non-steroidogenic COS-1 cells with P450c17 and p38 expression vectors showed that p38α, but not p38β, conferred 17,20 lyase activity on P450c17. Antiserum to P450c17 co-immunoprecipitated P450c17 and both p38 isoforms; however, knockdown of p38α, but not knockdown of p38β, inhibited 17,20 lyase activity in NCI-H295A cells. Bacterially expressed human P450c17 was phosphorylated by p38α in vitro at a non-canonical site, conferring increased 17,20 lyase activity. This phosphorylation increased the maximum velocity, but not the Michaelis constant, of the 17,20 lyase reaction. p38α phosphorylates P450c17 in a fashion that confers increased 17,20 lyase activity, implying that the production of adrenal androgens (adrenarche) is a regulated event.
Noncanonical Role of the PDZ4 Domain of the Adaptor Protein PDZK1 in the Regulation of the Hepatic High Density Lipoprotein Receptor Scavenger Receptor Class B, Type I (SR-BI)*

Background: PDZK1 (four PDZ domains) regulates the hepatic HDL receptor SR-BI. Results: PDZK1's PDZ2 and PDZ3 domains are not required, whereas PDZ4 is, possibly because PDZ4 mediates membrane binding. Conclusion: Regulation of SR-BI via PDZK1's PDZ domains is complex. Significance: Combined canonical (target peptide binding) and noncanonical (peptide binding-independent) PDZ domain functions can result in optimal activity of a PDZ domain-containing adaptor protein. The four PDZ (PDZ1 to PDZ4) domain-containing adaptor protein PDZK1 controls the expression, localization, and function of the HDL receptor scavenger receptor class B, type I (SR-BI), in hepatocytes in vivo. This control depends on both the PDZ4 domain and the binding of SR-BI's cytoplasmic C terminus to the canonical peptide-binding sites of either the PDZ1 or PDZ3 domain (no binding to PDZ2 or PDZ4). Using transgenic mice expressing in the liver domain deletion (ΔPDZ2 or ΔPDZ3), domain replacement (PDZ2→1), or target peptide binding-negative (PDZ4(G389P)) mutants of PDZK1, we found that neither PDZ2 nor PDZ3 nor the canonical target peptide binding activity of PDZ4 were necessary for hepatic SR-BI regulatory activity. Immunohistochemical studies established that the localization of PDZK1 on hepatocyte cell surface membranes in vivo is dependent on its PDZ4 domain and the presence of SR-BI. Analytical ultracentrifugation and hydrogen deuterium exchange mass spectrometry suggested that the requirement of PDZ4 for localization and SR-BI regulation is not due to PDZ4-mediated oligomerization or induction of conformational changes in the PDZ123 portion of PDZK1. However, surface plasmon resonance analysis showed that PDZ4, but not the other PDZ domains, can bind vesicles that mimic the plasma membrane. Thus, PDZ4 may potentiate PDZK1's regulation of SR-BI by promoting its lipid-mediated attachment to the cytoplasmic membrane. Our results show that not all of the PDZ domains of a multi-PDZ domain-containing adaptor protein are required for its biological activities and that both canonical target peptide binding and noncanonical (peptide binding-independent) capacities of PDZ domains may be employed by a single such adaptor for optimal in vivo activity.
Androgens and innate immunity in rehabilitated semi‐captive orangutans (Pongo pygmaeus morio) from Malaysian Borneo

Despite the implications for the development of life‐history traits, endocrine–immune trade‐offs in apes are not well studied. This is due, in part, to difficulty in sampling wild primates, and lack of methods available for immune measures using samples collected noninvasively. Evidence for androgen‐mediated immune trade‐offs in orangutans is virtually absent, and very little is known regarding their pattern of adrenal development and production of adrenal androgens. To remedy both of these deficiencies, sera were collected from orangutans (Pongo pygmaeus morio) (N = 38) at the Sepilok Orangutan Rehabilitation Centre, Sabah, Malaysia, during routine health screenings. Testosterone, dehydroepiandrosterone (DHEA), and dehydroepiandrosterone‐sulfate (DHEA‐S) were assayed, along with two measures of functional innate immunity. DHEA‐S concentrations, but not DHEA, increased with age in this sample of 1–18 year old animals. DHEA concentrations were higher in animals with higher levels of serum bacteria killing ability, while DHEA‐S and testosterone concentrations were higher in animals with reduced complement protein activity. Patterns of DHEA‐S concentration in this sample are consistent with patterns of adrenarche observed in other apes. Results from this study suggest that in addition to testosterone, DHEA and DHEA‐S may have potent effects on immunological activity in this species. Am. J. Primatol. 77:642–650, 2015. © 2015 Wiley Periodicals, Inc.
Exercise testing in the equine athlete

Summary In the last decade there have been significant technological advances and increases in the availability of portable diagnostic equipment for use in the assessment of athletic performance. The design and application of appropriate exercise tests are critical factors for the diagnosis of conditions affecting equine performance and it is therefore necessary for veterinary surgeons using such equipment to develop an understanding of appropriate exercise testing methods. This is the first in a series of three papers covering exercise testing in the equine athlete, interpretation of heart rate and rhythm during exercise and interpretation of dynamic upper respiratory tract function. In this first paper, both performance and clinical exercise testing in the equine athlete are reviewed.
Identifying potentially unhealthy housing conditions in Rosengård – a cross-sectional study in immigrant households with small children in Sweden

The social determinants of health form a complex and dynamic web, shaping both population and individual health and disease. The built environment and especially human dwellings influence health in many ways. Associations between factors such as dampness, mould growth, heating/insulation, and crowding on the one hand and respiratory health and infectious diseases on the other hand have been studied in several populations. In Sweden, social and health inequalities have slowly been increasing. The linkages between them have however not been characterized in detail. In this cross-sectional study, the apartments of families of children with respiratory symptoms living in a socially disadvantaged immigrant neighbourhood in Malmo, in which the incidence of bad housing was known to be high, were assessed for potentially harmful housing-related exposures. This information was correlated with subjective questionnaire-based assessments of dampness and mould in the apartments. While agreement between objective and subjective assessments was generally only fair, the questions performed better at excluding relevant exposures. The utility of screening questions related to housing is related to the prevalence of the exposure in the population and to the default plan of action. A case is made for interdisciplinary home visits to be the default option in families of children with respiratory symptoms. The overall prevalence of other contributory factors that are potentially harmful to health was high in this population; potentially protective factors were distributed unevenly as well. Factors related to the built environment cannot and should not be seen in isolation, as they often co-vary with other social determinants of health. Housing is however an important pathway along which social inequalities translate into health inequalities.
Why do orangutans leave the trees? Terrestrial behavior among wild Bornean orangutans (Pongo pygmaeus wurmbii) at Tuanan, Central Kalimantan

Orangutans (genus Pongo) are the largest arboreal mammals, but Bornean orangutans (P. pygmaeus spp.) also spend time on the ground. Here, we investigate ground use among orangutans using 32,000 hr of direct focal animal observations from a well‐habituated wild population of Bornean orangutans (P. p. wurmbii) living in a closed‐canopy swamp forest at Tuanan, Central Kalimantan, Indonesia. Ground use did not change with increasing observation time of well‐habituated individuals, suggesting it was not an artifact of observer presence. Flanged males spent the most time on the ground (ca. 5% of active time), weaned immatures the least (around 1%). Females and immatures descended mainly to feed, especially on termites, whereas flanged males traveled more while on the ground. Flanged males may travel more inconspicuously, and perhaps also faster, when moving on the ground. In addition, orangutans engaged in ground‐specific behavior, including drinking from and bathing in swamp pools. Supplementary records from 20 ground‐level camera traps, totaling 3986 trap days, confirmed the observed age‐sex biases in ground use at Tuanan. We conclude that ground use is a natural part of the Bornean orangutan behavioral repertoire, however it remains unclear to what extent food scarcity and canopy structure explain population differences in ground use. Am. J. Primatol. 77:1216–1229, 2015. © 2015 Wiley Periodicals, Inc.
Family caregiving in chronic illness: Alzheimer's disease, cancer, heart disease, mental illness, and stroke.

Overview of Family Caregiving Theoretical Perspectives on Caregiving Caregiving in Cancer Caregiving in Heart Disease Caregiving in Stroke Caregiving in Alzheimer's Disease Caregiving in Chronic Mental Illness Common Factors Affecting Family Caregivers of the Chronically Ill A Research Synthesis Caregiver Intervention Outcome Studies Future Directions Research and Policy
The Structure of the Ternary Complex of Krev Interaction Trapped 1 (KRIT1) Bound to Both the Rap1 GTPase and the Heart of Glass (HEG1) Cytoplasmic Tail*

Background: KRIT1 FERM domain can bind Rap1 GTPase and HEG1 cytoplasmic tail. Results: We solved the crystal structure of the KRIT1-Rap1-HEG1 ternary complex and show how KRIT1 F2 subdomain contributes to Rap1 binding specificity. Conclusion: The structure rationalizes the capacity of KRIT1 to form this ternary complex. Significance: We reveal the structure of Rap1 in complex with HEG1-bound KRIT1 and identify a novel determinant of Rap1 specificity. Loss of function mutation in Krev interaction trapped 1 (KRIT1) causes autosomal dominant familial cerebral cavernous malformations and disrupts cardiovascular development. The biological function of KRIT1 requires that its FERM (band 4.1, ezrin, radixin, moesin) domain physically interact with both the small GTPase Rap1 and the cytoplasmic tail of the Heart of glass (HEG1) membrane anchor. In this study, we show that the KRIT1 FERM domain can bind both Rap1 and HEG1 simultaneously, and we solved the crystal structure of the KRIT1-Rap1-HEG1 ternary complex. Rap1 binds on the surface of the F1 and F2 subdomains, in an interaction that leaves its Switch II region accessible to other potential effectors. HEG1 binds in a hydrophobic pocket at the KRIT1 F1 and F3 interface, and there is no overlap with the Rap1-binding site. Indeed, the affinity of KRIT1 or the KRIT1-Rap1 complex for HEG1 is comparable (Kd = 1.2 and 0.96 μm, respectively) showing that there is no competition between the two sites. Furthermore, analysis of this structure revealed a specific ionic interaction between the F2 lobe of KRIT1 and Rap1 that could explain the remarkable Rap1 specificity of KRIT1. This structural insight enabled design of KRIT1(K570I), a mutant that binds Rap1 with 8-fold lower affinity and exhibits increased binding to HRas. These data show that HEG1 can recruit the Rap1-KRIT complex to the plasma membrane where Rap1's Switch II region remains accessible and reveals an important determinant of KRIT1's specificity for Rap1.
A Secondary Structural Transition in the C-helix Promotes Gating of Cyclic Nucleotide-regulated Ion Channels*

Background: Cyclic nucleotide-regulated channels are involved in sensory transduction and repetitive electrical activity. Results: Transition metal ion FRET and electrophysiology demonstrated a coil-to-helix transition within the ligand binding domain. Conclusion: Agonist binding stabilized the structure of the C-helix, triggering channel gating. Significance: This transition is important for ion channel activation and may be necessary for the activation of other cyclic nucleotide-regulated proteins. Cyclic nucleotide-regulated ion channels bind second messengers like cAMP to a C-terminal domain, consisting of a β-roll, followed by two α-helices (B- and C-helices). We monitored the cAMP-dependent changes in the structure of the C-helix of a C-terminal fragment of HCN2 channels using transition metal ion FRET between fluorophores on the C-helix and metal ions bound between histidine pairs on the same helix. cAMP induced a change in the dimensions of the C-helix and an increase in the metal binding affinity of the histidine pair. cAMP also caused an increase in the distance between a fluorophore on the C-helix and metal ions bound to the B-helix. Stabilizing the C-helix of intact CNGA1 channels by metal binding to a pair of histidines promoted channel opening. These data suggest that ordering of the C-helix is part of the gating conformational change in cyclic nucleotide-regulated channels.
A Chrysin Derivative Suppresses Skin Cancer Growth by Inhibiting Cyclin-dependent Kinases*

Background: Binding to the ATP site results in poor selectivity; therefore, development of ATP-noncompetitive inhibitors is needed. Results: A modified chrysin with anticancer activity targets Cdks and binds to a Cdk2 allosteric site, not the ATP pocket. Conclusion: Modified chrysin is a novel ATP-noncompetitive inhibitor. Significance: This pharmacophore model might provide insights for the development of new ATP-noncompetitive agents. Chrysin (5,7-dihydroxyflavone), a natural flavonoid widely distributed in plants, reportedly has chemopreventive properties against various cancers. However, the anticancer activity of chrysin observed in in vivo studies has been disappointing. Here, we report that a chrysin derivative, referred to as compound 69407, more strongly inhibited EGF-induced neoplastic transformation of JB6 P+ cells compared with chrysin. It attenuated cell cycle progression of EGF-stimulated cells at the G1 phase and inhibited the G1/S transition. It caused loss of retinoblastoma phosphorylation at both Ser-795 and Ser-807/811, the preferred sites phosphorylated by Cdk4/6 and Cdk2, respectively. It also suppressed anchorage-dependent and -independent growth of A431 human epidermoid carcinoma cells. Compound 69407 reduced tumor growth in the A431 mouse xenograft model and retinoblastoma phosphorylation at Ser-795 and Ser-807/811. Immunoprecipitation kinase assay results showed that compound 69407 attenuated endogenous Cdk4 and Cdk2 kinase activities in EGF-stimulated JB6 P+ cells. Pulldown and in vitro kinase assay results indicated that compound 69407 directly binds with Cdk2 and Cdk4 in an ATP-independent manner and inhibited their kinase activities. A binding model between compound 69407 and a crystal structure of Cdk2 predicted that compound 69407 was located inside the Cdk2 allosteric binding site. The binding was further verified by a point mutation binding assay. Overall results indicated that compound 69407 is an ATP-noncompetitive cyclin-dependent kinase inhibitor with anti-tumor effects, which acts by binding inside the Cdk2 allosteric pocket. This study provides new insights for creating a general pharmacophore model to design and develop novel ATP-noncompetitive agents with chemopreventive or chemotherapeutic potency.
Bioactive factors in milk across lactation: Maternal effects and influence on infant growth in rhesus macaques (Macaca mulatta)

Among mammals, numerous bioactive factors in milk vary across mothers and influence offspring outcomes. This emerging area of research has primarily investigated such dynamics within rodent biomedical models, domesticated dairy breeds, and among humans in clinical contexts. Less understood are signaling factors in the milk of non‐human primates. Here, we report on multiple bioactive components in rhesus macaque (Macaca mulatta) milk and their associations with maternal and infant characteristics. Milk samples were collected from 59 macaques at multiple time points across lactation in conjunction with maternal and infant morphometrics and life‐history animal records. Milk was assayed for adiponectin (APN), epidermal growth factor (EGF) and its receptor (EGF‐R), and transforming growth factor beta 2 (TGF‐β2). Regression models were constructed to assess the contributions of maternal factors on variation in milk bioactives, and on the relationship of this variation to infant body mass and growth. Maternal body mass, parity, social rank, and infant sex were all predictive of concentrations of milk bioactives. Primiparous mothers produced milk with higher adiponectin, but lower EGF, than multiparous mothers. Heavier mothers produced milk with lower EGF and EGF‐R, but higher TGF‐β2. Mothers of daughters produced milk with higher TGF‐β2. Mid‐ranking mothers produced milk with higher mean EGF and adiponectin concentrations than low‐ranking mothers. Milk EGF and EGF‐R were positively associated with infant body mass and growth rate. Importantly, these signaling bioactives (APN, EGF, EGF‐R, and TGF‐β2) were significantly correlated with nutritional values of milk. The effects of milk signals remained after controlling for the available energy in milk revealing the added physiological role of non‐nutritive milk bioactives in the developing infant. Integrating analyses of energetic and other bioactive components of milk yields an important perspective for interpreting the magnitude, sources, and consequences of inter‐individual variation in milk synthesis. Am. J. Primatol. 78:838–850, 2016. © 2016 Wiley Periodicals, Inc.
Online peer support for patients with somatic diseases

With the availability of the Internet, so rose the opportunity to share concerns and experiences with peers online. In this thesis the meaning of online support groups for patients with rheumatoid arthritis, fibromyalgia and breast cancer was examined from different perspectives. To this aim, six studies were performed. In study 1 a content analysis was conducted on 1500 messages derived from various discussion groups for the aforementioned conditions. The main aim was to examine if the disadvantages that are mentioned in literature, actually occur. In study 2 a group of participants of various online discussion groups were (qualitatively and quantitatively) questioned about their use of these groups, the perceived advantages and disadvantages and if and how their participation empowers them. In the third study a representative sample of patients, derived from 2 hospitals, were asked to fill out written questionnaires to examine their use of Internet for health-related reasons and their participation in face-to-face and online support groups. The factors that are associated to this were also examined. In study 4 a group of patients with rheumatoid arthritis was qualitatively questioned about reasons for (not) engaging in (online) peer-to-peer support. In the fifth study a survey was sent to all Dutch rheumatologists and oncologists to explore their experiences and attitudes with regard to their patients’ health-related Internet use. Finally, in study 6, 23 webmasters of online support groups were interviewed to explore their reasons for initiating online support groups and the factors that determine the success of these groups. The main finding of our studies was that patients who use online support groups benefit in various ways, however, only a small proportion of Dutch patients participate in these groups.
Mutational and Structural Analysis of the Tandem Zinc Finger Domain of Tristetraprolin*

Background: Structural aspects of tristetraprolin (TTP) binding to mRNA are poorly understood. Results: RNA binding can be differentially affected by sequence-equivalent residues within the tandem zinc fingers of TTP. Conclusion: Residues at sequence-equivalent positions within each zinc finger may not be structurally or functionally equivalent. Significance: Different TTP family members may have structural features that dictate unique modes of interacting with RNA. Tristetraprolin (TTP), the best known member of a class of tandem (R/K)YKTELCX8CX5CX3H zinc finger proteins, can destabilize target mRNAs by first binding to AU-rich elements (AREs) in their 3′-untranslated regions (UTRs) and subsequently promoting deadenylation and ultimate destruction of those mRNAs. This study sought to determine the roles of selected amino acids in the RNA binding domain, known as the tandem zinc finger (TZF) domain, in the ability of the full-length protein to bind to AREs within the tumor necrosis factor α (TNF) mRNA 3′-UTR. Within the CX8C region of the TZF domain, mutation of some of the residues specific to TTP, not found in other members of the TTP protein family, resulted in decreased binding to RNA as well as inhibited mRNA deadenylation and decay. Evaluation of simulation solution models revealed a distinct structure in the second zinc finger of TTP that was induced by the presence of these TTP-specific residues. In addition, mutations within the lead-in sequences preceding the first C of highly conserved residues within the CX5C or CX3H regions or within the linker region between the two fingers also perturbed both RNA binding and the simulation model of the TZF domain in complex with RNA. We conclude that, although the majority of conserved residues within the TZF domain of TTP are required for productive binding, not all residues at sequence-equivalent positions in the two zinc fingers of the TZF domain of TTP are functionally equivalent.