Miracle-dz/UniFAR-FTUs huggingface dataset:
Maternal Deprivation in Rats is Associated with Corticotrophin‐Releasing Hormone (CRH) Promoter Hypomethylation and Enhances CRH Transcriptional Responses to Stress in Adulthood

Exposure to stress during early development causes long‐lasting alterations in behaviour and hypothalamic pituitary adrenal (HPA) axis activity, including increased expression of corticotrophin‐releasing hormone (CRH). To determine whether early‐life stress causes epigenetic changes in the CRH promoter leading to increased CRH transcription, 8‐week old female and male rats, subjected to maternal deprivation (MD) between days 2 and 13 post‐birth, were studied for HPA axis responses to stress and CRH promoter methylation in the hypothalamic paraventricular nucleus (PVN) and central nucleus of the amygdala (CeA). Plasma corticosterone and PVN CRH heteronuclear (hn)RNA responses to acute restraint stress were higher in MD rats of both sexes. DNA methylation analysis of the CRH promoter revealed a significantly lower percentage of methylation in two CpGs preceding (CpG1) and inside (CpG2) the cyclic AMP‐response element (CRE) at −230 bp in the CRH promoter in the PVN but not the CeA of MD rats. Gel‐shift assays, using nuclear proteins from forskolin‐treated hypothalamic 4B cells and CRH promoter CRE oligonucleotides, unmethylated or methylated at CpG1, revealed a strong band that was supershifted by phospho‐cAMP response element‐binding antibody. This band was 50% weaker using oligonucleotides methylated at CpG2 (intra‐CRE), or methylated at both CpG1 and CpG2. These findings demonstrate that HPA axis hypersensitivity caused by neonatal stress causes long‐lasting enhanced CRH transcriptional activity in the PVN of both sexes. Hypomethylation of the CRH promoter CRE, a region critical for CRH transcriptional activation, could serve as a mechanism for the increased transcriptional responses to stress observed in MD rats.
Type I Interferon Controls Propagation of Long Interspersed Element-1*

Background: Type 1 interferons (IFN1) mediate defense against viruses but their role in regulating retrotransposon activities is unknown. Results: LINE-1 retrotransposon induces IFN1, which in turn inhibits LINE-1 retrotransposition. Conclusion: IFN1 regulate activities and propagation of LINE-1. Significance: Given that retrotransposons alter the genome, IFN1 play a role in maintenance of genomic integrity. Type I interferons (IFN) including IFNα and IFNβ are critical for the cellular defense against viruses. Here we report that increased levels of IFNβ were found in testes from mice deficient in MOV10L1, a germ cell-specific RNA helicase that plays a key role in limiting the propagation of retrotransposons including Long Interspersed Element-1 (LINE-1). Additional experiments revealed that activation of LINE-1 retrotransposons increases the expression of IFNβ and of IFN-stimulated genes. Conversely, pretreatment of cells with IFN suppressed the replication of LINE-1. Furthermore, the efficacy of LINE-1 replication was increased in isogenic cell lines harboring inactivating mutations in diverse elements of the IFN signaling pathway. Knockdown of the IFN receptor chain IFNAR1 also stimulated LINE-1 propagation in vitro. Finally, a greater accumulation of LINE-1 was found in mice that lack IFNAR1 compared with wild type mice. We propose that LINE-1-induced IFN plays an important role in restricting LINE-1 propagation and discuss the putative role of IFN in preserving the genome stability.
How active are young adult women

Issues addressed: 1) Whether young adult women with and without young children are sufficiently active for health benefit. 2) Whether physical activity (PA) measures used in national surveys capture the everyday activities performed by young adult women. Methods: Fifteen mothers (M) with at least one child under 5 years and 19 women of similar age without young children (W) were recruited. Each participant completed a self report survey assessing frequency and duration of PA associated with walking to and from places, active leisure or recreation, and paid and non-paid work or household chores. After completing the survey, participants were given an activity diary and a pedometer to record their daily activities and daily number of steps during two typical weekdays and one typical weekend. Results: There were no significant differences between M and W for number of steps (9563 vs 9789), PA energy expenditure (PAEE) calculated from the diary (2047 vs 1918 METmins/day), and PAEE estimated from the survey responses (1545 vs 1500 METmins/week). Housework and laundry, shopping, and gardening accounted for about 15% of weekly PAEE (from diaries), while walking and recreation/leisure accounted for 6-9%. Weekend and weekly pedometer steps were significantly associated with PAEE estimated from the diary (r=0.50 and 0.31 respectively); however, the level of agreement between pedometer steps and the survey outcomes was poor (-0.03 to 0.20). Conclusions: The young women in this study engaged in substantial amounts of physical activity. Survey instruments that assess predominantly transport and leisure-time activity do not however adequately capture the activities performed by young women as part of daily living. So what: Better measures of physical activity that allow for the estimation of PAEE from tasks performed by women in the home and workplace are required. (author abstract)
Organizational factors that support the implementation of a nursing best practice guideline.

BACKGROUND The context of the healthcare setting may play a crucial role in influencing the implementation of best practice guidelines in nursing. Further study is required to understand these organizational factors. Two variables, organizational culture and leadership, are thought to influence the adoption of best practice guidelines. AIM A discussion of organizational factors that influence best practice guideline adoption is presented. A small pilot study is provided as an example of methods for further research. METHODS A quantitative survey of nursing staff was conducted. RESULTS Results from the pilot study reveal variability in best practice guideline implementation despite the presence of a culture of organizational learning and transformational leadership. CONCLUSIONS There is beginning evidence in the literature that culture and leadership are key elements influencing guideline implementation. In this pilot work on two inpatient units where a nursing best practice guideline was implemented, a supportive organizational culture and key people leading change were present. Implications for further studies are offered. IMPLICATIONS FOR NURSING MANAGEMENT Nursing leaders interested in promoting the use of best practice guidelines must pay attention to the organizational context in which nursing care occurs. A supportive culture where learning is valued coupled with transformational leadership may be key factors in the implementation and the sustainability of best practice guidelines.
Structure-guided Mutational Analysis of Gene Regulation by the Bacillus subtilis pbuE Adenine-responsive Riboswitch in a Cellular Context*

Background: Riboswitches regulate purine biosynthesis and transport in many bacteria. Results: Mutagenic analysis of an adenine-responsive riboswitch revealed features important for efficient co-transcriptional regulation. Conclusion: Adenine binding to the riboswitch results in a local barrier to strand exchange by the transcriptional terminator, whose formation is facilitated by a stem-loop element. Significance: This is the first study comprehensively analyzing an RNA structural switch in a cellular context. Riboswitches are a broadly distributed form of RNA-based gene regulation in Bacteria and, more rarely, Archaea and Eukarya. Most often found in the 5′-leader sequence of bacterial mRNAs, they are generally composed of two functional domains: a receptor (aptamer) domain that binds an effector molecule and a regulatory domain (or expression platform) that instructs the expression machinery. One of the most studied riboswitches is the Bacillus subtilis adenine-responsive pbuE riboswitch, which regulates gene expression at the transcriptional level, up-regulating expression in response to increased intracellular effector concentrations. In this work, we analyzed sequence and structural elements that contribute to efficient ligand-dependent regulatory activity in a co-transcriptional and cellular context. Unexpectedly, we found that the P1 helix, which acts as the antitermination element of the switch in this RNA, supported ligand-dependent activation of a reporter gene over a broad spectrum of lengths from 3 to 10 bp. This same trend was also observed using a minimal in vitro single-turnover transcription assay, revealing that this behavior is intrinsic to the RNA sequence. We also found that the sequences at the distal tip of the terminator not directly involved in alternative secondary structure formation are highly important for efficient regulation. These data strongly support a model in which the switch is highly localized to the P1 helix adjacent to the ligand-binding pocket that likely presents a local kinetic block to invasion of the aptamer by the terminator.
Long‐term effects of pancreatic transplantation on diabetic neuropathy

Restoration of a long‐lasting euglycemic state by a functioning pancreatic transplantation (PTx) is the most logical treatment for insulin‐dependent diabetes mellitus and for amelioration of secondary complications, including neuropathy. We evaluated neurological function by clinical examination, nerve conduction studies, and autonomic function tests in 115 patients with a functioning PTx and in 92 control patients treated with insulin, at baseline and 1, 2, 3.5, 5, 7, and 10 years later. In control patients, neuropathy progressively worsened during follow‐up. The clinical examination score and composite indices of abnormality of motor and sensory nerve conduction decreased significantly at all intervals tested. Autonomic function indices also decreased, but significantly only after 1 year. In patients who received a successful PTx the neuropathy improved. The motor and sensory nerve conduction indices increased significantly at all intervals after transplantation, whereas the clinical examination and autonomic tests improved only slightly. Patients who received either a PTx alone, a PTx after a kidney graft, or simultaneous pancreatic and kidney transplantations improved similarly over the follow‐up. These results indicate that a functioning PTx halts the progression and improves the signs of diabetic polyneuropathy by restoration of a normoglycemic state.
A Pilot Study Evaluating Care Staffs' Perceptions of their Experience of Psychological Consultation within a Mental Health Setting

The aim of this study was to conduct a preliminary evaluation of a mental health consultation process provided to staff working in residential care settings for young people. Using a semi-structured interview approach, qualitative information was gathered from six participants about their experiences of the consultation. A thematic analysis of the interviews generated four key themes, which appear to reflect the different stages of consultation. These included: initiating consultation; building the consultative relationship; overcoming obstacles; and seeing the value of consultation. These themes, along with prevalent sub-themes, are discussed with reference to the effectiveness of the consultation process and the implications for use of psychological consultation in clinical practice. Implications for future practice include providing information to potential consultees before the consultation process, being accessible informally to the consultee and adopting a variety of approaches based on the consultee's needs. The limitations of this study and recommendations for future research are also discussed.
Ubiquitin-conjugating Enzyme Cdc34 and Ubiquitin Ligase Skp1-Cullin-F-box Ligase (SCF) Interact through Multiple Conformations*

Background: The ubiquitin-conjugating enzyme Cdc34 rapidly associates with ubiquitin ligase SCF to achieve processive ubiquitination. Results: Protein cross-linking was used to capture native Cdc34-SCF complexes. Conclusion: The acidic tail domain of Cdc34 interacts with the basic canyon region on SCF in multiple conformations. Significance: Multiple conformations between Cdc34 and SCF enable their rapid association. In the ubiquitin-proteasome system, protein substrates are degraded via covalent modification by a polyubiquitin chain. The polyubiquitin chain must be assembled rapidly in cells, because a chain of at least four ubiquitins is required to signal for degradation, and chain-editing enzymes in the cell may cleave premature polyubiquitin chains before achieving this critical length. The ubiquitin-conjugating enzyme Cdc34 and ubiquitin ligase SCF are capable of building polyubiquitin chains onto protein substrates both rapidly and processively; this may be explained at least in part by the atypically fast rate of Cdc34 and SCF association. This rapid association has been attributed to electrostatic interactions between the acidic C-terminal tail of Cdc34 and a feature on SCF called the basic canyon. However, the structural aspects of the Cdc34-SCF interaction and how they permit rapid complex formation remain elusive. Here, we use protein cross-linking to demonstrate that the Cdc34-SCF interaction occurs in multiple conformations, where several residues from the Cdc34 acidic tail are capable of contacting a broad region of the SCF basic canyon. Similar patterns of cross-linking are also observed between Cdc34 and the Cul1 paralog Cul2, implicating the same mechanism for the Cdc34-SCF interaction in other members of the cullin-RING ubiquitin ligases. We discuss how these results can explain the rapid association of Cdc34 and SCF.
A Conserved Interaction That Is Essential for the Biogenesis of Histone Locus Bodies*

Background: Proteins involved in expression of histone genes co-localize to discrete nuclear structures called histone locus bodies (HLBs). Results: Two main components of HLBs directly interact with each other through their C-terminal regions. Conclusion: Formation of HLBs depends on a subset of critical protein-protein interactions. Significance: Our work may help to understand the mechanisms that integrate transcription of histone genes with maturation of the nascent histone transcripts. Nuclear protein, ataxia-telangiectasia locus (NPAT) and FLICE-associated huge protein (FLASH) are two major components of discrete nuclear structures called histone locus bodies (HLBs). NPAT is a key co-activator of histone gene transcription, whereas FLASH through its N-terminal region functions in 3′ end processing of histone primary transcripts. The C-terminal region of FLASH contains a highly conserved domain that is also present at the end of Yin Yang 1-associated protein-related protein (YARP) and its Drosophila homologue, Mute, previously shown to localize to HLBs in Drosophila cells. Here, we show that the C-terminal domain of human FLASH and YARP interacts with the C-terminal region of NPAT and that this interaction is essential and sufficient to drive FLASH and YARP to HLBs in HeLa cells. Strikingly, only the last 16 amino acids of NPAT are sufficient for the interaction. We also show that the C-terminal domain of Mute interacts with a short region at the end of the Drosophila NPAT orthologue, multi sex combs (Mxc). Altogether, our data indicate that the conserved C-terminal domain shared by FLASH, YARP, and Mute recognizes the C-terminal sequence of NPAT orthologues, thus acting as a signal targeting proteins to HLBs. Finally, we demonstrate that the C-terminal domain of human FLASH can be directly joined with its N-terminal region through alternative splicing. The resulting 190-amino acid MiniFLASH, despite lacking 90% of full-length FLASH, contains all regions necessary for 3′ end processing of histone pre-mRNA in vitro and accumulates in HLBs.
The MicroRNA-130/301 Family Controls Vasoconstriction in Pulmonary Hypertension*

Background: The microRNA-130/301 family regulates pulmonary hypertension (PH), but its breadth of activity remains undefined. Results: Predicted by network analysis, microRNA-130/301 members regulate vasoactive factors such as endothelin-1 for pulmonary vascular cross-talk. Conclusion: The microRNA-130/301 family promotes vasoconstriction in PH. Significance: This microRNA-based mechanism of vascular cross-talk is central to the systems-wide actions of microRNA-130/301 in PH. Pulmonary hypertension (PH) is a complex disorder, spanning several known vascular cell types. Recently, we identified the microRNA-130/301 (miR-130/301) family as a regulator of multiple pro-proliferative pathways in PH, but the true breadth of influence of the miR-130/301 family across cell types in PH may be even more extensive. Here, we employed targeted network theory to identify additional pathogenic pathways regulated by miR-130/301, including those involving vasomotor tone. Guided by these predictions, we demonstrated, via gain- and loss-of-function experimentation in vitro and in vivo, that miR-130/301-specific control of the peroxisome proliferator-activated receptor γ regulates a panel of vasoactive factors communicating between diseased pulmonary vascular endothelial and smooth muscle cells. Of these, the vasoconstrictive factor endothelin-1 serves as an integral point of communication between the miR-130/301-peroxisome proliferator-activated receptor γ axis in endothelial cells and contractile function in smooth muscle cells. Thus, resulting from an in silico analysis of the architecture of the PH disease gene network coupled with molecular experimentation in vivo, these findings clarify the expanded role of the miR-130/301 family in the global regulation of PH. They further emphasize the importance of molecular cross-talk among the diverse cellular populations involved in PH.
Adherencia al Tratamiento basado en Comportamientos en Pacientes Diabéticos Cartagena de Indias, Colombia

Objetivo Conocer los niveles de adherencia auto-reportada y control metabolico, interpretados desde un enfoque biopsicosocial, con el fin de proponer estrategias culturalmente adaptadas que permitan el logro de mejores resultados en los programas impartidos. Metodos se realizo un estudio descriptivo que incluyo 131 pacientes diabeticos usuarios de una empresa promotora de salud (EPS) de la ciudad de Cartagena de Indias, Colombia. Se aplicaron entrevistas para conocer los niveles de adherencia auto-reportada y se midio hemoglobina glucosilada A1C para evaluar control metabolico. Resultados En una escala de 0 a 7, el puntaje promedio de adherencia obtenido fue de 4,6 (IC 95 % 3,8-5,4). Los mayores puntajes fueron 6,6 para no tabaquismo (IC 95 % 5,5-7,8) y 6,2 para medicacion (IC 95 % 5,9-6,6). Los menores fueron 0,4 para automonitoreo (IC 95 % 0,4-0,5); 3,4 para ejercicio fisico (IC 95 % 2,8-4,0) y 4,4 para consumo de vegetales (IC 95 % 3,7-5,2). El control metabolico deseable se logro en el 58 % de los pacientes, pero no guardo relacion con la adherencia auto-reportada (p>0,05). Conclusiones Se recomienda evaluar la adherencia de manera independiente para cada comportamiento, fortalecer los pilares no farmacologicos del tratamiento en los programas dirigidos a pacientes diabeticos e implementar las acciones pertinentes para lograr mayores niveles de accesibilidad a estos.
Wound healing : alternatives in management

Part 1 The wound healing environment: normal healing nutrition and wound healing factoring affecting repair bacteriology and infection control growth factors, extracellular matrix and wound healing. Part 2 Evaluation: methods of wound evaluation. Part 3 Principles and techniques: wound debridement and irrigation dressings electrical stimulation for wound healing adjunctive interventions for wound healing. Part 4 Case management and documentation: management of the insensate foot management of wounds secondary to vascular disease prevention and treatment of wounds cause by pressure documentation for reimbursement.
Mechanisms of Membrane Binding of Small GTPase K-Ras4B Farnesylated Hypervariable Region*

Background: K-Ras4B modulates downstream signaling at different lipid microdomains. Results: K-Ras4B farnesyl group spontaneously inserts into the disordered lipid microdomains, but phosphorylation prohibits the farnesyl membrane insertion. Conclusion: The farnesyl may determine K-Ras4B function in different membrane microdomain environments. Significance: Figuring out K-Ras4B localization at different membrane microdomains is important for a more complete understanding Ras-effector interactions mediating signaling pathways. K-Ras4B belongs to a family of small GTPases that regulates cell growth, differentiation and survival. K-ras is frequently mutated in cancer. K-Ras4B association with the plasma membrane through its farnesylated and positively charged C-terminal hypervariable region (HVR) is critical to its oncogenic function. However, the structural mechanisms of membrane association are not fully understood. Here, using confocal microscopy, surface plasmon resonance, and molecular dynamics simulations, we observed that K-Ras4B can be distributed in rigid and loosely packed membrane domains. Its membrane binding domain interaction with phospholipids is driven by membrane fluidity. The farnesyl group spontaneously inserts into the disordered lipid microdomains, whereas the rigid microdomains restrict the farnesyl group penetration. We speculate that the resulting farnesyl protrusion toward the cell interior allows oligomerization of the K-Ras4B membrane binding domain in rigid microdomains. Unlike other Ras isoforms, K-Ras4B HVR contains a single farnesyl modification and positively charged polylysine sequence. The high positive charge not only modulates specific HVR binding to anionic phospholipids but farnesyl membrane orientation. Phosphorylation of Ser-181 prohibits spontaneous farnesyl membrane insertion. The mechanism illuminates the roles of HVR modifications in K-Ras4B targeting microdomains of the plasma membrane and suggests an additional function for HVR in regulation of Ras signaling.
Pathogenic Gene Mutations or Variants Identified by Targeted Gene Sequencing in Adults With Hemophagocytic Lymphohistiocytosis

Hemophagocytic lymphohistiocytosis (HLH) can be classified into primary HLH and secondary HLH. Primary HLH usually occurs in infants and children with an underlying genetic defect, and there are also teens and occasional adults with primary HLH. Most cases with secondary HLH are adult patients with secondary triggers including infections, malignancies, and autoimmune diseases. The distinction between primary HLH and secondary HLH seems to be less straightforward, as patients with secondary HLH may also have genetic defects while primary HLH can be triggered by secondary causes. In this study, using amplicon-based targeted gene sequencing (TGS), we sequenced eighteen HLH-related genes in 112 adult HLH cases, which were mostly secondary HLH. Mutations or rare variants were identified in 48 cases (42.9%). All the variants except one were missense variants, and biallelic gene mutations were identified in 3 cases in which only one case harbored homogenous missense mutation. Recurrent variants including UNC13D p.G863D and AP3B1 p.T359A are much more prevalent in our cohort than in normal East Asian population, and in silico analysis predicted pathogenicity of these variants. In conclusion, according to our study, genetic defects may also contribute to the development of adult HLH cases or secondary HLH cases.