bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lung tissue bioenergetics and caspase activity in rodents
Authors: Alsuwaidi Ahmed R , Alsamri Mohammed T , Alfazari Ali S , Almarzooqi Saeeda , Albawardi Alia , Othman Aws R , Pramathan Thachillath , Hartwig Stacey M , Varga Steven M , Souid Abdul-Kader 
Published: January 2013

Abstract

Background
This study aimed to establish a suitable in vitro system for investigating effects of respiratory pathogens and toxins on lung tissue bioenergetics (cellular respiration and ATP content) and caspase activity. Wistar rats and C57Bl/6 mice were anesthetized by sevoflurane inhalation. Lung fragments were then collected and incubated at 37°C in a continuously gassed (with 95% O2:5% CO2) Minimal Essential Medium (MEM) or Krebs-Henseleit buffer. Phosphorescence O2 analyzer that measured dissolved O2 concentration as a function of time was used to monitor the rate of cellular mitochondrial O2 consumption. Cellular ATP content was measured using the luciferin/luciferase system. The caspase-3 substrate N-acetyl-asp-glu-val-asp-7-amino-4-methylcoumarin (Ac-DEVD-AMC) was used to monitor intracellular caspase activity; cleaved AMC moieties (reflecting caspase activity) were separated on HPLC and detected by fluorescence. Lung histology and immunostaining with anti-cleaved caspase-3 antibody were also performed.

Results
For Wistar rats, the values of k

c
and ATP for 0 < t ≤ 7 h (mean ± SD) were 0.15 ± 0.02 μM O2 min-1 mg-1 (n = 18, coefficient of variation, Cv = 13%) and 131 ± 69 pmol mg-1 (n = 16, Cv = 53%), respectively. The AMC peak areas remained relatively small despite a ~5-fold rise over 6 h. Good tissue preservation was evident despite time-dependent increases in apoptotic cells. Lung tissue bioenergetics, caspase activity and structure were deleterious in unoxygenated or intermittently oxygenated solutions. Incubating lung tissue in O2 depleted MEM for 30 min or anesthesia by urethane had no effect on lung bioenergetics, but produced higher caspase activity.

Conclusions
Lung tissue bioenergetics and structure could be maintained in vitro in oxygenated buffer for several hours and, thus, used as biomarkers for investigating respiratory pathogens or toxins.
Treatment of penetrating trauma of the extremities: ten years’ experience at a dutch level 1 trauma center
Authors: Van Waes Oscar JF , Van Lieshout Esther MM , Hogendoorn Wouter , Halm Jens A , Vermeulen Jefrey 
Published: January 2013

Abstract

Background
A selective non-operative management (SNOM) has found to be an adequate and safe strategy to assess and treat patients suffering from penetrating trauma of the extremities (PTE). With this SNOM comes a strategy in which adjuvant investigations or interventions are not routinely performed, but based on physical examination only.

Methods
All subsequent patients presented with PTE at a Dutch level I trauma center from October 2000 to June 2011 were included in this study. In-hospital and long-term outcome was analysed in the light of assessment of these patients according to the SNOM protocol.

Results
A total of 668 patients (88.2% male; 33.8% gunshot wounds) with PTE presented at the Emergency Department of a level 1 traumacenter, of whom 156 were admitted for surgical treatment or observation. Overall, 22 (14%) patients that were admitted underwent exploration of the extremity for vascular injury. After conservative observation, two (1.5%) patients needed an intervention to treat (late onset) vascular complications. Other long-term extremity related complications were loss of function or other deformity (n = 9) due to missed nerve injury, including 2 patients with peroneal nerve injury caused by delayed compartment syndrome treatment.

Conclusion
A SNOM protocol for initial assessment and treatment of PTE is feasible and safe. Clinical examination of the injured extremity is a reliable diagnostic 'tool' for excluding vascular injury. Repeated assessments for nerve injuries are important as these are the ones that are frequently missed and result in long-term disability. Level of evidence: II / III, retrospective prognostic observational cohort study Key words Penetrating trauma, extremity, vascular injury, complications.
Vascular disrupting agent for neovascular age related macular degeneration: a pilot study of the safety and efficacy of intravenous combretastatin a-4 phosphate
Authors: Ibrahim Mohamed A , Do Diana V , Sepah Yasir J , Shah Syed M , Van Anden Elizabeth , Hafiz Gulnar , Donahue J Kevin , Rivers Richard , Balkissoon Jai , Handa James T , Campochiaro Peter A , Nguyen Quan Dong 
Published: January 2013

AbstractBackgroundThis study was designed to assess the safety, tolerability, and efficacy of intravenous infusion of CA4P in patients with neovascular age-related macular degeneration (AMD).MethodsProspective, interventional, dose-escalation clinical trial. Eight patients with neovascular AMD refractory to at least 2 sessions of photodynamic therapy received CA4P at a dose of 27 or 36 mg/m2as weekly intravenous infusion for 4 consecutive weeks. Safety was monitored by vital signs, ocular and physical examinations, electrocardiogram, routine laboratory tests, and collection of adverse events. Efficacy was assessed using retinal fluorescein angiography, optical coherence tomography, and best corrected visual acuity (BCVA).ResultsThe most common adverse events were elevated blood pressure (46.7%), QTc prolongation (23.3%), elevated temperature (13.3%), and headache (10%), followed by nausea and eye injection (6.7%). There were no adverse events that were considered severe in intensity and none resulted in discontinuation of treatment. There was reduction of the excess foveal thickness by 24.15% at end of treatment period and by 43.75% at end of the two-month follow-up (p = 0.674 and 0.161, respectively). BCVA remained stable throughout the treatment and follow-up periods.ConclusionsThe safety profile of intravenous CA4P was consistent with that reported in oncology trials of CA4P and with the class effects of vascular disruptive agents; however, the frequency of adverse events was different. There are evidences to suggest potential efficacy of CA4P in neovascular AMD. However, the level of systemic safety and efficacy indicates that systemic CA4P may not be suitable as an alternative monotherapy to current standard-of-care therapy.Trial registrationClinicalTrials.gov NCT01570790.
Resuscitative strategies in traumatic hemorrhagic shock
Authors: Bouglé Adrien , Harrois Anatole , Duranteau Jacques 
Published: January 2013

Abstract
Managing trauma patients with hemorrhagic shock is complex and difficult. Despite our knowledge of the pathophysiology of hemorrhagic shock in trauma patients that we have accumulated during recent decades, the mortality rate of these patients remains high. In the acute phase of hemorrhage, the therapeutic priority is to stop the bleeding as quickly as possible. As long as this bleeding is uncontrolled, the physician must maintain oxygen delivery to limit tissue hypoxia, inflammation, and organ dysfunction. This process involves fluid resuscitation, the use of vasopressors, and blood transfusion to prevent or correct acute coagulopathy of trauma. The optimal resuscitative strategy is controversial. To move forward, we need to establish optimal therapeutic approaches with clear objectives for fluid resuscitation, blood pressure, and hemoglobin levels to guide resuscitation and limit the risk of fluid overload and transfusion.
Reversible impairment of the ku80 gene by a recyclable marker in Aspergillus aculeatus
Authors: Tani Shuji , Tsuji Atsushi , Kunitake Emi , Sumitani Jun-ichi , Kawaguchi Takashi 
Published: January 2013

Abstract
Auxotrophic mutants of Aspergillus can be isolated in the presence of counter-selective compounds, but the process is laborious. We developed a method to enable reversible impairment of the ku80 gene (Aaku80) in the imperfect fungus Aspergillus aculeatus. Aaku80 was replaced with a selection marker, orotidine 5’-phosphate decarboxylase (pyrG), followed by excision of pyrG between direct repeats (DR) to yield the Aaku80 deletion mutant (MR12). The gene-targeting efficiency at the ornithine carbamoyltransferase (argB) locus was drastically elevated from 3% to 96% in MR12. The frequency of marker recycling depended on DR length. One uridine auxotroph was obtained from 3.3 × 105, 1.4 × 105, and 9.2 × 103 conidia from strains harboring 20-, 98-, and 495-bp DRs, respectively. Because these strains maintained the short DRs after 5 d of cultivation, we investigated whether Aaku80 function was disrupted by pyrG insertion with the 20-bp DR and restored after excision of pyrG. The Aaku80 disruption mutant (coku80) was bred by inserting pyrG sandwiched between 20-bp DRs into the second intron of Aaku80, followed by excision of pyrG between the DRs to yield the coku80rec strain. Analyses of homologous recombination frequency and methyl methanesulfonate sensitivity demonstrated that Aaku80 function was disrupted in coku80 but restored in coku80rec. Furthermore, pyrG was maintained in coku80 at least for ten generations. These data indicated that reversible impairment of ku80 in A. aculeatus is useful for functional genomics in cases where genetic segregation is not feasible.
Expression, purification and characterization of a recombinant fusion protein based on the human papillomavirus-16 E7 antigen
Authors: Granadillo Milaid , Batte Aileen , Lugo Victoria M , Musacchio Alexis , Bequet-Romero Mónica , Betancourt Lázaro , Besada Vladimir , Javier Luis , Molina Raychel , Falcón Viviana , Torrens Isis 
Published: January 2013

AbstractA fusion protein comprising a cell penetrating and immunostimulatory peptide corresponding to residues 32 to 51 of theLimulus polyphemusprotein linked to human papillomavirus (HPV)-16 E7 antigen (LALF32-51-E7) was expressed inE. coliBL21 (DE3) cells. The recombinant protein inE. coliaccounted for approximately 18% of the total cellular protein and purified with a single affinity chromatographic step. Yields of approximately 38 mg purified LALF32-51-E7 per liter of induced culture was obtained with an overall 52% recovery and constitutes a promising setting for the future production and scaling-up. Purified protein was characterized as soluble aggregates with molecular weight larger than 670 kDa, which is considered an important property to increase the immunogenicity of an antigen preparation. The recombinant fusion protein LALF32-51-E7 will be a promising vaccine candidate for the treatment of HPV-16 related malignancies.
Effect of humidity on egg hatchability and reproductive biology of the bamboo borer (Dinoderus minutus Fabricius)
Authors: Norhisham Ahmad R , Abood Faizah , Rita Muhamad , Hakeem Khalid Rehman 
Published: January 2013

Abstract
Wood products are highly exposed to infestation by powder post beetles. Dinoderus minutus (bamboo borer) is a wood boring beetle that seriously damage dried bamboo and finished bamboo products. Management of D. minutus using pesticides showed negative effects on environment despite being very costly. By understanding influence of natural climatic conditions on their reproductive behaviour, could help us to develop a cost effective and environmental friendly strategy to cope up with this problem. In the present study, reproductive parameters and egg development of the bamboo borer were determined at 20%, 40%, 56%, 75% and 85% r.h. levels at constant temperature of 30° ± 2°C with 8 L-16D photoregime. From the results, eclosion to first instar larva was recorded at all relative humidities tested. The lowest shortest percentage of hatchability was recorded at 20% and 85% relative humidity with a mean incubation period of 4.63 ± 0.25 and 10.43 ± 0.32 days, respectively. It was noted that pre-ovipositional period decreased from 14.20 ± 0.49 to 7.20 ± 0.31 days as relative humidity increased from 20% to 75% and slightly increased to 8.00 ± 0.37 days at 85% relative humidity. We conclude that female beetles may have a particular hygropreference in oviposition as total egg production increased with increasing relative humidity.
Biogenic amines activate blood leukocytes via trace amine-associated receptors TAAR1 and TAAR2
Authors: Babusyte Agne , Kotthoff Matthias , Fiedler Julia , Krautwurst Dietmar 
Published: January 2013

Abstract
TAAR1 and TAAR2 are necessary for biogenic amine-induced in vitro activation of PMN migration, Th-cell cytokine secretion, and B-cell immunoglobulin E production.
Certain biogenic amines, such as 2-PEA, TYR, or T1AM, modulate blood pressure, cardiac function, brain monoaminergic systems, and olfaction-guided behavior by specifically interacting with members of a group of rhodopsin-like receptors, TAAR. A receptor that is absent from olfactory epithelia but had long been identified in the brain and a variety of peripheral tissues, TAAR1 has been found recently in blood B cells, suggesting a functional role of TAAR1 in these cells. With the present study, we have set out to clarify the expression and functional roles of TAAR in different isolated human blood leukocyte types. Here, we report the functional expression of TAAR1 and its closest relative TAAR2 in blood PMN and T and B cells. Both receptors are coexpressed in a subpopulation of PMN, where they are necessary for the chemosensory migration toward the TAAR1 ligands 2-PEA, TYR, and T1AM, with EC50 values of 0.43 ± 0.05 nM, 0.52 ± 0.05 nM, and 0.25 ± 0.04 nM, respectively. The same amines, with similar potencies, triggered cytokine or Ig secretion, in purified blood T or B cells, respectively. Notably, 2-PEA regulated mRNA expression of 28 T cell function-related genes, above all of the CCL5. In siRNA-guided experiments, TAAR1 and TAAR2 proved to be necessary for amine-induced blood leukocyte functions. In summary, our results demonstrate that biogenic amines potently regulate blood cell functions via TAAR1 and TAAR2 and open the perspective of their specific pharmacological modulation.
Research Resource: Tissue- and Pathway-Specific Metabolomic Profiles of the Steroid Receptor Coactivator (SRC) Family
Authors: York Brian , Sagen Jørn V. , Tsimelzon Anna , Louet Jean-Francios , Chopra Atul R. , Reineke Erin L. , Zhou Suoling , Stevens Robert D. , Wenner Brett R. , Ilkayeva Olga , Bain James R. , Xu Jianming , Hilsenbeck Susan G. , Newgard Christopher B. , O'Malley Bert W. 
Published: January 2013

AbstractThe rapidly growing family of transcriptional coregulators includes coactivators that promote transcription and corepressors that harbor the opposing function. In recent years, coregulators have emerged as important regulators of metabolic homeostasis, including the p160 steroid receptor coactivator (SRC) family. Members of the SRC family have been ascribed important roles in control of gluconeogenesis, fat absorption and storage in the liver, and fatty acid oxidation in skeletal muscle. To provide a deeper and more granular understanding of the metabolic impact of the SRC family members, we performed targeted metabolomic analyses of key metabolic byproducts of glucose, fatty acid, and amino acid metabolism in mice with global knockouts (KOs) of SRC-1, SRC-2, or SRC-3. We measured amino acids, acyl carnitines, and organic acids in five tissues with key metabolic functions (liver, heart, skeletal muscle, brain, plasma) isolated from SRC-1, -2, or -3 KO mice and their wild-type littermates under fed and fasted conditions, thereby unveiling unique metabolic functions of each SRC. Specifically, SRC-1 ablation revealed the most significant impact on hepatic metabolism, whereas SRC-2 appeared to impact cardiac metabolism. Conversely, ablation of SRC-3 primarily affected brain and skeletal muscle metabolism. Surprisingly, we identified very few metabolites that changed universally across the three SRC KO models. The findings of this Research Resource demonstrate that coactivator function has very limited metabolic redundancy even within the homologous SRC family. Furthermore, this work also demonstrates the use of metabolomics as a means for identifying novel metabolic regulatory functions of transcriptional coregulators.
InDrosophila, RhoGEF2 cooperates with activated Ras in tumorigenesis through a pathway involving Rho1–Rok–Myosin-II and JNK signalling
Authors: Khoo Peytee , Allan Kirsten , Willoughby Lee , Brumby Anthony M. , Richardson Helena E. 
Published: January 2013

SummaryThe Ras oncogene contributes to ~30% of human cancers, but alone is not sufficient for tumorigenesis. In a Drosophila screen for oncogenes that cooperate with an activated allele of Ras (RasACT) to promote tissue overgrowth and invasion, we identified the GTP exchange factor RhoGEF2, an activator of Rho-family signalling. Here, we show that RhoGEF2 also cooperates with an activated allele of a downstream effector of Ras, Raf (RafGOF). We dissect the downstream pathways through which RhoGEF2 cooperates with RasACT (and RafGOF), and show that RhoGEF2 requires Rho1, but not Rac, for tumorigenesis. Furthermore, of the Rho1 effectors, we show that RhoGEF2 + Ras (Raf)-mediated tumorigenesis requires the Rho kinase (Rok)–Myosin-II pathway, but not Diaphanous, Lim kinase or protein kinase N. The Rho1–Rok–Myosin-II pathway leads to the activation of Jun kinase (JNK), in cooperation with RasACT. Moreover, we show that activation of Rok or Myosin II, using constitutively active transgenes, is sufficient for cooperative tumorigenesis with RasACT, and together with RasACT leads to strong activation of JNK. Our results show that Rok–Myosin-II activity is necessary and sufficient for Ras-mediated tumorigenesis. Our observation that activation of Myosin II, which regulates Filamentous actin (F-actin) contractility without affecting F-actin levels, cooperates with RasACT to promote JNK activation and tumorigenesis, suggests that increased cell contractility is a key factor in tumorigenesis. Furthermore, we show that signalling via the Tumour necrosis factor (TNF; also known as Egr)-ligand–JNK pathway is most likely the predominant pathway that activates JNK upon Rok activation. Overall, our analysis highlights the need for further analysis of the Rok–Myosin-II pathway in cooperation with Ras in human cancers.
Use of zinc-finger nucleases to knock out theWASgene in K562 cells: a human cellular model for Wiskott-Aldrich syndrome
Authors: Toscano Miguel G. , Anderson Per , Muñoz Pilar , Lucena Gema , Cobo Marién , Benabdellah Karim , Gregory Philip D. , Holmes Michael C. , Martin Francisco 
Published: January 2013

SummaryMutations in the WAS gene cause Wiskott-Aldrich syndrome (WAS), which is characterized by eczema, immunodeficiency and microthrombocytopenia. Although the role of WASP in lymphocytes and myeloid cells is well characterized, its role on megakaryocyte (MK) development is poorly understood. In order to develop a human cellular model that mimics the megakaryocytic-derived defects observed in WAS patients we used K562 cells, a well-known model for study of megakaryocytic development. We knocked out the WAS gene in K562 cells using a zinc-finger nuclease (ZFN) pair targeting the WAS intron 1 and a homologous donor DNA that disrupted WASP expression. Knockout of WASP on K562 cells (K562WASKO cells) resulted in several megakaryocytic-related defects such as morphological alterations, lower expression of CD41α, lower increments in F-actin polymerization upon stimulation, reduced CD43 expression and increased phosphatidylserine exposure. All these defects have been previously described either in WAS-knockout mice or in WAS patients, validating K562WASKO as a cell model for WAS. However, K562WASPKO cells showed also increased basal F-actin and adhesion, increased expression of CD61 and reduced expression of TGFβ and Factor VIII, defects that have never been described before for WAS-deficient cells. Interestingly, these phenotypic alterations correlate with different roles for WASP in megakaryocytic differentiation. All phenotypic alterations observed in K562WASKO cells were alleviated upon expression of WAS following lentiviral transduction, confirming the role of WASP in these phenotypes. In summary, in this work we have validated a human cellular model, K562WASPKO, that mimics the megakaryocytic-related defects found in WAS-knockout mice and have found evidences for a role of WASP as regulator of megakaryocytic differentiation. We propose the use of K562WASPKO cells as a tool to study the molecular mechanisms involved in the megakaryocytic-related defects observed in WAS patients and as a cellular model to study new therapeutic strategies.
Effect of an environmental health educational programme for paediatricians in an Egyptian University Hospital: before and after study
Authors: Abbas Reem A , Alghobashy Ashgan A 
Published: January 2013

Objectives To study the effect of an educational intervention on paediatricians’ knowledge, attitudes and practices regarding children's environmental health and to identify the sources of information and common environmental history taking constrains. Design Before and after study. Setting Zagazig University Paediatric Hospital. Participants Practising paediatricians from all specialty units. Main outcome measures The outcome of a specifically designed educational programme about paediatric environmental health was assessed using structured pre- and post-test questionnaires. Results Nearly half the participants were aware about most of the paediatric environmental health-related topics. Textbooks/guidelines (85.7%) and the Internet (64.3%) were the main sources of information. The participants demonstrated relatively strong positive attitudes towards the importance of children's environmental health. However, less than half of them (44.6%) reported environmental history taking as a routine practice; where lack of time (94.6%), wide range of hazardous exposures (91.1%) and lack of expertise and training (91.1%) were the main constrains. Significant improvement in participants’ knowledge, attitudes and practices was revealed after the educational programme. Conclusions There is a demand for continuous medical education about environmental health in paediatric practice, particularly environmental and occupational history taking.