Real-Time Killing Assays to Assess the Potency of a New Anti-Simian Immunodeficiency Virus Chimeric Antigen Receptor T Cell.
The success of chimeric antigen receptor (CAR) T cell therapies for treating leukemia has resulted in a booming interest for the technology. Expression of a CAR in T cells allows redirection of their natural cytolytic activity toward cells presenting a specific designated surface antigen. Although CAR T cell therapies have thus far shown promising results mostly in B cell malignancy trials, interest in their potential to treat other diseases is on the rise, including using CAR T cells to control human immunodeficiency virus infection. The assessment of CAR T cell potency toward specific targets in vitro is a critical preclinical step. In this study, we describe novel assays that monitor the cytotoxicity of candidate CAR T cells toward simian immunodeficiency virus (SIV) infected CD4 T cells. The assays involve live cell imaging using a fluorescence microscopy system that records in real time the disappearance or appearance of targets infected with SIV carrying a fluorescent protein gene. The assays are highly reproducible, and their rapid turn around and reduced cost present a significant advance regarding the efficient preclinical evaluation of CAR T cell constructs and are broadly applicable to potential human diseases that could benefit from CAR T cell therapy.


Nonmotor symptoms in genetic Parkinson disease.
To review current knowledge on nonmotor symptoms (NMS), particularly psychiatric features, in genetic Parkinson disease (PD) and to provide original data for genetic and idiopathic PD. A MEDLINE search using Parkinson and known PD genes focused on the presence of depression, anxiety, hallucinations, and dementia was performed. Original data from 82 outpatients with idiopathic (n = 55) and genetic (n = 27) PD were obtained. All studies including information on NMS and patients with genetic PD. Study methods and clinical and genetic information were summarized. The literature search yielded 1855 citations; 305 included genetic information on PD patients, of which 119 also contained information on any type of NMS (990 cases). Availability of information varied by gene and type of NMS; studies differed by recruitment and examination method. Literature search and original data showed high frequencies of the following NMS: depression, 8% to 37% (literature) and 33% to 40% (our data); anxiety, 7% to 37% (literature) and 10% to 22% (our data); hallucinations, 3% to 23% (literature) and 23% to 29% (our data); and dementia, 5% to 26% (literature), absent in our own data. Data on NMS in genetic PD are limited. Specific data needs include a systematic approach to NMS assessment reporting permitting comparability of studies. Overall, the frequency of NMS in genetic PD does not appear to be higher and may even be lower than in idiopathic PD. Nonmotor symptoms have a high impact on the patients' quality of life and caregiver burden and should be considered important and often treatable concomitant features of genetic PD.


LayNii: A software suite for layer-fMRI.
High-resolution fMRI in the sub-millimeter regime allows researchers to resolve brain activity across cortical layers and columns non-invasively. While these high-resolution data make it possible to address novel questions of directional information flow within and across brain circuits, the corresponding data analyses are challenged by MRI artifacts, including image blurring, image distortions, low SNR, and restricted coverage. These challenges often result in insufficient spatial accuracy of conventional analysis pipelines. Here we introduce a new software suite that is specifically designed for layer-specific functional MRI: LayNii. This toolbox is a collection of command-line executable programs written in C/C++ and is distributed opensource and as pre-compiled binaries for Linux, Windows, and macOS. LayNii is designed for layer-fMRI data that suffer from SNR and coverage constraints and thus cannot be straightforwardly analyzed in alternative software packages. Some of the most popular programs of LayNii contain 'layerification' and columnarization in the native voxel space of functional data as well as many other layer-fMRI specific analysis tasks: layer-specific smoothing, model-based vein mitigation of GE-BOLD data, quality assessment of artifact dominated sub-millimeter fMRI, as well as analyses of VASO data.


Comprehensive Analysis of the Soybean (Glycine max) GmLAX Auxin Transporter Gene Family.
The phytohormone auxin plays a critical role in regulation of plant growth and development as well as plant responses to abiotic stresses. This is mainly achieved through its uneven distribution in plant via a polar auxin transport process. Auxin transporters are major players in polar auxin transport. The AUXIN RESISTENT 1/LIKE AUX1 (AUX/LAX) auxin influx carriers belong to the amino acid permease family of proton-driven transporters and function in the uptake of indole-3-acetic acid (IAA). In this study, genome-wide comprehensive analysis of the soybean AUX/LAX (GmLAX) gene family, including phylogenic relationships, chromosome localization, and gene structure, was carried out. A total of 15 GmLAX genes, including seven duplicated gene pairs, were identified in the soybean genome. They were distributed on 10 chromosomes. Despite their higher percentage identities at the protein level, GmLAXs exhibited versatile tissue-specific expression patterns, indicating coordinated functioning during plant growth and development. Most GmLAXs were responsive to drought and dehydration stresses and auxin and abscisic acid (ABA) stimuli, in a tissue- and/or time point- sensitive mode. Several GmLAX members were involved in responding to salt stress. Sequence analysis revealed that promoters of GmLAXs contained different combinations of stress-related cis-regulatory elements. These studies suggest that the soybean GmLAXs were under control of a very complex regulatory network, responding to various internal and external signals. This study helps to identity candidate GmLAXs for further analysis of their roles in soybean development and adaption to adverse environments.


[Pilot study of The National Child and Adolescent Health Survey: sampling design].
The concept of a nation-wide Health Survey for Children and Adolescents has been tested over one year in a pilot study with 1,630 participating children and adolescents. A major methodological aspect of this pilot phase was to compare different variants of study sample recruitment regarding their feasibility. There was a choice between a sample originating from the population registry and a cluster sample of schools and school classes for the age group from 11 to 15 years. As a result, after weighing the advantages and disadvantages of both sampling designs, a continuous population registry sample has been recommended. The acquired knowledge and experience will be translated into reality in 2003 when the main course of the survey, which will include about 18,000 children and adolescents, will start.


[History of processing of Sojae Semen Praeparatum].
Sojae Semen Praeparatum (SSP) is commonly used as a type of dietetic Chinese herb. By collecting and analyzing ancient and recent literatures, a textual criticism was conducted on the historical evolution of the processing of SSP. Fermented soybean was recorded in Shijing, and relevant rational processing was described in Qimin Yaoshu. In the early time, fermented soybean included the type of "salty" and "light". After the Ming Dynasty, "light" fermented soybean or SSP was recognized as a better medicinal matter than salty fermented soybean, and the fermentation processing was recorded more clearly. In modern time, many characteristic methods for processing SSP have been developed. Today, the processing of SSP is mainly based on the Chinese Pharmacopoeia, which records soybean as a main ingredient and Artemisiae Annuae Herba, Mori Folium as excipients.


Complete Sequences of Six IncA/C Plasmids of Multidrug-Resistant Salmonella enterica subsp. enterica Serotype Newport.
Multidrug-resistant (MDR) Salmonella enterica subsp. enterica serotype Newport has been a long-standing public health concern in the United States. We present the complete sequences of six IncA/C plasmids from animal-derived MDR S. Newport ranging from 80.1 to 158.5 kb. They shared a genetic backbone with S. Newport IncA/C plasmids pSN254 and pAM04528.


Anaerobic treatment of army ammunition production wastewater containing perchlorate and RDX.
Perchlorate is an oxidizer that has been routinely used in solid rocket motors by the Department of Defense and National Aeronautics and Space Administration. Royal Demolition Explosive (RDX) is a major component of military high explosives and is used in a wide variety of munitions. Perchlorate bearing wastewater typically results from production of solid rocket motors, while RDX is transferred to Army industrial wastewaters during load, assemble and pack operations for new munitions, and hot water or steam washout for disposal and deactivation of old munitions (commonly referred to as demilitarization, or simply demil). Biological degradation in Anaerobic Fluidized Bed Reactors (AFBR), has been shown to be an effective method for the removal of both perchlorate and RDX in contaminated wastewater. The focus of this study was to determine the effectiveness of removal of perchlorate and RDX, individually and when co-mingled, using ethanol as an electron donor under steady state conditions. Three AFBRs were used to assess the effectiveness of this process in treating the wastewater. The performance of the bioreactors was monitored relative to perchlorate, RDX, and chemical oxygen demand removal effectiveness. The experimental results demonstrated that the biodegradation of perchlorate and RDX was more effective in bioreactors receiving the single contaminant than in the bioreactor where both contaminants were fed.


In vitro evaluation of aerosol delivery of aztreonam lysine (AZLI): an adult mechanical ventilation model.
The delivery profile of Aztreonam lysine (AZLI) during mechanical ventilation (MV) is unknown. We evaluated the amount of AZLI drug delivered using an in vitro model of adult MV. An adult lung model designed to mimic current clinical practice was used. Both nebulizers were placed before a Y-piece and 4 settings were tested: A) Aeroneb solo® [AS] with a t-piece; B) AS with the spacer; C) M-Neb® [MN] with a t-piece and D) MN with the spacer. Performance was evaluated in terms of: 1) Mass median aerodynamic diameter (MMAD); 2) Geometric standard deviation (GSD), 3) Fine particle dose (FPD), 4) Fine particle fraction (FPF), 5) Inhalable mass (IM), and 6) Recovery rate (RR). Both devices showed an adequate delivery of AZLI during MV, with MMAD between 2.4-2.5 µm and 87% of FPF. The FPD (38.8 and 31.7), IM (44.8 and 36.1) and RR (30 and 24) were similar for AS and MN respectively. Nebulizer aerosol delivery increased (50 and 70% respectively) for both nebulizers when using the spacer. Both AS and MN showed a good aerosol delivery profile for AZLI during in vitro mechanical ventilation. Better aerosol delivery performance was obtained using the spacer.


Pharmacological stress testing.
Pharmacological stress in conjunction with radionuclide myocardial perfusion imaging has become a widely used noninvasive method of assessing patients with known or suspected coronary artery disease. In the United States, over one third of perfusion imaging studies are performed with pharmacological stress. Pharmacological stress agents fall into two categories: coronary vasodilating agents such as dipyridamole and adenosine, and cardiac positive inotropic agents such as dobutamine and arbutamine. For both, in the presence of coronary artery disease (CAD), perfusion image abnormalities result from heterogeneity of coronary blood flow reserve. Vasodilating agents work directly on the coronary vessels to increase blood flow, whereas inotropic agents work indirectly by increasing myocardial work load, which then leads to an increase in coronary blood flow. Both classes of agents have high accuracies for diagnosing coronary artery disease, and they have excellent safety records with acceptably low occurrences of side effects. For dipyridamole planar thallium imaging, pooled analysis yields a sensitivity of 85% and a specificity of 87% for diagnosis of coronary disease, but there is a large variation in reported values depending on various factors, such as the extent of postcatheterization referral bias, the type of imaging (planar versus single photon emission computed tomography [SPECT]), the types of patients being studied (single versus multivessel disease, men versus women), and the imaging agent used (thallium versus one of the technetium-based agents). Diagnostic accuracies for adenosine are similar to those of dipyridamole, with reported overall sensitivities ranging from 83% to 97%, and specificities ranging from 38% to 94%. For dobutamine, pooled analyses yield a sensitivity of 82% and a specificity of 75%. There is some concern that dobutamine may interfere with uptake of technetium-99m sestamibi, lowering the sensitivity for detection of disease, and thus the vasdodilating agents are generally preferred. Pharmacological stress testing has high clinical use for risk stratifying patients with known or suspected CAD, in patients after myocardial infarction, and in patients needing noncardiac surgery. Vasodilating agents are particularly advantageous in assessing post-myocardial infarction patients, allowing testing as soon as 2 days after the event. Like patients undergoing exercise stress testing, patients with normal perfusion images by pharmacological stress have a <1% annual incidence of cardiac events. The likelihood of an event increases with the extent and severity of perfusion abnormalities. However, it is important to consider clinical variables when using perfusion imaging for risk stratification, particularly in the presurgery patients. As with exercise testing, adjunct markers such as ST segment depression during testing, lung uptake of radiotracer (if thallium is used), and ventricular cavity dilatation add additional prognostic information to that available from the perfusion images alone. The aim of current research is to find better agents that are easier to use and that have fewer side effects. MRE-0470 is an experimental vasodilating agent that is more receptor selective than adenosine and promises a lower incidence of hypotension. Arbutamine more closely simulates exercise than dobutamine, and it can be administered by a closed-loop computerized delivery device. Work is also underway to look at novel uses of pharmacological stress agents, such as acquiring gated SPECT images during dobutamine infusion to enhance detection of myocardial viability. With increasing use of noninvasive testing in elderly patients and in patients with comorbidities that preclude adequate exercise, pharmacological stress testing has become an indispensable tool for radionuclide myocardial perfusion imaging studies. A good understanding of pharmacological stress testing is essential for performing high-quality nuclear cardiology


Assessing the structural heterogeneity of supercooled liquids through community inference.
We present an information-theoretic approach inspired by distributional clustering to assess the structural heterogeneity of particulate systems. Our method identifies communities of particles that share a similar local structure by harvesting the information hidden in the spatial variation of two- or three-body static correlations. This corresponds to an unsupervised machine learning approach that infers communities solely from the particle positions and their species. We apply this method to three models of supercooled liquids and find that it detects subtle forms of local order, as demonstrated by a comparison with the statistics of Voronoi cells. Finally, we analyze the time-dependent correlation between structural communities and particle mobility and show that our method captures relevant information about glassy dynamics.


Ten common estate planning mistakes to avoid.
While most Americans are aware of the importance of estate planning, many fail to make the proper arrangements for their heirs. Problems can arise from a number of mistake, such as underestimating assets, relying solely on a will, or not planning for estate tax payments.


Role of mast cells in chronic stress induced colonic epithelial barrier dysfunction in the rat.
Stress may be an important factor in exacerbating inflammatory bowel disease but the underlying mechanism is unclear. Defective epithelial barrier function may allow uptake of luminal antigens that stimulate an immune/inflammatory response. Here, we examined the effect of chronic stress on colonic permeability and the participation of mast cells in this response. Mast cell deficient Ws/Ws rats and +/+ littermate controls were submitted to water avoidance stress or sham stress (one hour/day) for five days. Colonic epithelial permeability to a model macromolecular antigen, horseradish peroxidase, was measured in Ussing chambers. Epithelial and mast cell morphology was studied by light and electron microscopy. Chronic stress significantly increased macromolecular flux and caused epithelial mitochondrial swelling in +/+ rats, but not in Ws/Ws rats, compared with non-stressed controls. Stress increased the number of mucosal mast cells and the proportion of cells showing signs of activation in +/+ rats. No mast cells or ultrastructural abnormalities of the epithelium were present in Ws/Ws rats. Increased permeability in +/+ rats persisted for 72 hours after stress cessation. Chronic stress causes an epithelial barrier defect and epithelial mitochondrial damage, in parallel with mucosal mast cell hyperplasia and activation. The study provides further support for an important role for mast cells in stress induced colonic mucosal pathophysiology.


The fibroblast growth factor receptor genetic status as a potential predictor of the sensitivity to CH5183284/Debio 1347, a novel selective FGFR inhibitor.
The FGF receptors (FGFR) are tyrosine kinases that are constitutively activated in a subset of tumors by genetic alterations such as gene amplifications, point mutations, or chromosomal translocations/rearrangements. Recently, small-molecule inhibitors that can inhibit the FGFR family as well as the VEGF receptor (VEGFR) or platelet-derived growth factor receptor (PDGFR) family displayed clinical benefits in cohorts of patients with FGFR genetic alterations. However, to achieve more potent and prolonged activity in such populations, a selective FGFR inhibitor is still needed. Here, we report the identification of CH5183284/Debio 1347, a selective and orally available FGFR1, FGFR2, and FGFR3 inhibitor that has a unique chemical scaffold. By interacting with unique residues in the ATP-binding site of FGFR1, FGFR2, or FGFR3, CH5183284/Debio 1347 selectively inhibits FGFR1, FGFR2, and FGFR3 but does not inhibit kinase insert domain receptor (KDR) or other kinases. Consistent with its high selectivity for FGFR enzymes, CH5183284/Debio 1347 displayed preferential antitumor activity against cancer cells with various FGFR genetic alterations in a panel of 327 cancer cell lines and in xenograft models. Because of its unique binding mode, CH5183284/Debio 1347 can inhibit FGFR2 harboring one type of the gatekeeper mutation that causes resistance to other FGFR inhibitors and block FGFR2 V564F-driven tumor growth. CH5183284/Debio 1347 is under clinical investigation for the treatment of patients harboring FGFR genetic alterations.


[Dietetic treatment of hyperlipoproteinemias].
The authors mention the previous conditions to the prescription of a diet for primary hyperlipoproteinemia : definition of the metabolic disease and of its nutritional dependence, precise knowledge of earlier nutritional uses, demonstration of vascular risk factor linked to the hyperlipoproteinemia, i.e. obesity which always requires a hypocaloric diet. A low cholesterol and saturated fatty acid diet reduces by 10% the cholesterolemia, and sometimes exempts from use of medical drugs in moderate hypercholesterolemia. The exceptional hyperchylomicronemia are reduced by drastic reduction of the lipid fraction of the diet, which is compensated by use of MCT. The dietetic treatment of endogenous hypertriglyceridemia depends on their nutritional dependence : an alcohol dependence implies a complete suppression of alcoholic drinks. A glucid dependence implies the suppression of simple carbohydrates and a reduction of the glucidic fraction of the diet.


An E. coli gene product required for lambda site-specific recombination.
We report characteristics of himA mutations of E. coli, selected for their inability to support the site-specific recombination reaction involved in the formation of lysogens by bacteriophage lambda. The himA allele lies at minute 38 on the chromosome. Three noncomplementing and closely linked mutations define the himA locus; one is a nonsense mutation which shows that the gene product is a protein. HimA mutations reduce both lambda integrative and excisive site-specific recombination. Since dominance tests demonstrate that himA mutations are recessive, it is probable that the himA protein is either a necessary component for site-specific recombination or, alternatively, regulates the expression of such a function. HimA mutations exhibit pleiotropic effects. They reduce integration of phages that have different attachment specificities from lambda and inhibit the growth of phage mu. In addition, himA mutations reduce precise excision of integrated phage mu as well as Tn elements. This pleiotropy suggests that the role of himA protein is nonspecific. Since all of the processes affected by himA mutations ultimately rely on protein-DNA interactions, we suggest that himA protein may act in an auxillary manner to facilitate these interactions.


Spontaneous neurotransmission: an independent pathway for neuronal signaling?
Recent findings suggest that spontaneous neurotransmission is a bona fide pathway for interneuronal signaling that operates independent of evoked transmission via distinct presynaptic as well as postsynaptic substrates. This article will examine the role of spontaneous release events in neuronal signaling by focusing on aspects that distinguish this process from evoked neurotransmission, and evaluate the mechanisms that may underlie this segregation.


Dual-Energy CT in Musculoskeletal Imaging: What Is the Role Beyond Gout?
OBJECTIVE. Dual-energy CT (DECT) involves the acquisition of CT attenuation data at two different energy levels. In musculoskeletal imaging, gout detection is the most validated clinical indication for DECT. However, there is emerging evidence for several other clinical musculoskeletal applications, including metal artifact reduction and bone marrow imaging for traumatic, neoplastic, and inflammatory processes. CONCLUSION. In this article, we review the current role of DECT in musculoskeletal imaging, primarily focusing on nongout entities.


Peak experiences and the afterglow phenomenon: when and how do therapeutic effects of hallucinogens depend on psychedelic experiences?
Interest in the therapeutic potential of psychedelic substances has recently resumed. During an early phase of human psychedelic research, their therapeutic application in different pathologies had been suggested, and the first evidence for efficacy was provided. The range of recent clinical applications of psychedelics spans from cluster headaches and obsessive-compulsive disorder to addiction and the treatment of fear and anxiety in patients suffering from terminal illness, indicating potentially different therapeutic mechanisms. A variety of approaches in psychotherapy emphasize subjective experiences, such as so-called peak experiences or afterglow phenomena, as differentially mediating therapeutic action. This review aims to re-evaluate earlier and recent concepts of how psychedelic substances may exert beneficial effects. After a short outline of neurophenomenological aspects, we discuss different approaches to how psychedelics are used in psychotherapy. Finally, we summarize evidence for the relationship between subjective experiences and therapeutic success. While the distinction between pharmacological and psychological action obviously cannot be clear-cut, they do appear to contribute differently from each other when their effects are compared with regard to pathologies.


Agreement between experiment and hybrid DFT calculations for O--H bond dissociation enthalpies in manganese complexes.
Information on the accuracy of DFT functionals for redox reactions in transition metal systems is rather limited. To analyze the performance of some popular functionals for redox reactions in manganese systems, calculated O--H bond dissociation enthalpies for Mn-ligands in six different complexes are compared to experimental results. In this benchmark, B3LYP performs well with a mean absolute error of 3.0 kcal/mol. B98 gives similar results to B3LYP (error of 3.8 kcal/mol). B3LYP* gives lower O--H bond strengths than B3LYP and has a mean error of 5.0 kcal/mol. Compared to B98 and B3LYP, B3LYP* has an error trend for the manganese ligands that is more similar to the error for a free water molecule. The nonhybrid functional BLYP consistently and significantly underestimates the O--H bond strengths by approximately 20 kcal/mol. HCTH407 has a rather large mean error of 9.4 kcal/mol and shows no consistent trend. The results support the use of hybrid functionals and the present computational method for large model systems containing manganese. An example is the oxygen evolving complex in photosystem II where hybrid functionals predict the appearance of a Mn(IV)-oxyl radical before the O--O bond formation step.


An effective strategy of magnetic stem cell delivery for spinal cord injury therapy.
Spinal cord injury (SCI) is a condition that results in significant mortality and morbidity. Treatment of SCI utilizing stem cell transplantation represents a promising therapy. However, current conventional treatments are limited by inefficient delivery strategies of cells into the injured tissue. In this study, we designed a magnetic system and used it to accumulate stem cells labelled with superparamagnetic iron oxide nanoparticles (SPION) at a specific site of a SCI lesion. The loading of stem cells with engineered SPIONs that guarantees sufficient attractive magnetic forces was achieved. Further, the magnetic system allowed rapid guidance of the SPION-labelled cells precisely to the lesion location. Histological analysis of cell distribution throughout the cerebrospinal channel showed a good correlation with the calculated distribution of magnetic forces exerted onto the transplanted cells. The results suggest that focused targeting and fast delivery of stem cells can be achieved using the proposed non-invasive magnetic system. With future implementation the proposed targeting and delivery strategy bears advantages for the treatment of disease requiring fast stem cell transplantation.


Oral lichenoid drug reaction associated with antihypertensive and hypoglycemic drugs.
Oral lichen planus and oral lichenoid drug reactions have similar clinical and histologic findings. The onset of oral lichenoid drug reactions appears to correspond to the administration of medications, especially antihypertensive drugs, oral hypoglycemic drugs, antimalarial drugs, gold salts, penicillamine and others. The author reports the case of 58-year-old male patient with oral lichenoid drug reaction, hypertension and diabetes mellitus. The oral manifestation showed radiated white lines with erythematous and erosive areas. The patient experienced pain and a burning sensation when eating spicy food. A tissue biopsy was carried out and revealed the characteristics of lichen planus. The patient was treated with 0.1% fluocinolone acetonide in an orabase as well as the replacement of the oral hypoglycemic and antihypertensive agents. The lesions improved and the burning sensation disappeared in two weeks after treatment. No recurrence was observed in the follow-up after three months.


Ozone, but not nitrogen dioxide, exposure decreases glutathione peroxidases in epithelial lining fluid of human lung.
Antioxidants, such as glutathione peroxidases (GPxs), in epithelial lining fluid (ELF) protect against health effects of oxidant pollutants, which includes O(3) or NO(2). We hypothesized that GPxs concentration in ELF is responsive to O(3) or NO(2) exposure. Subjects underwent two 4-h exposures to O(3) (0.22 ppm) and one to air. In another experiment, subjects underwent 3-h exposures to air and NO(2) (0.6 and 1.5 ppm). Bronchoalveolar lavage (BAL) was performed immediately or 18 h after O(3) exposure and 3.5 h after each NO(2) exposure. GPx activity and extracellular GPx (eGPx) protein concentrations were determined in ELF, and their relationships to markers of lung function, inflammation, and epithelial permeability were examined. Although the total amounts were not changed, basal (air) GPx activity (223.6 +/- 24.4 mU/ml), basal eGPx protein concentration (2.62 +/- 0.25 microg/ml), and basal ELF dilution factor (152.3 +/- 8.4) decreased 40% immediately after O(3) exposure and remained 30% decreased 18 h after exposure (p = 0.0001). No effect of NO(2) exposure on GPxs concentration was detected. There was an inverse correlation between baseline ELF eGPx protein concentration and the change in PMN 18 h after O(3) exposure (p = 0.04). Thus, O(3), a strong oxidant, decreases both GPx activity and eGPx protein in ELF, whereas NO(2), a weaker oxidant, does not. eGPx in ELF may protect against O(3)-induced airway inflammation.


Accurate vertical ionization energy and work function determinations of liquid water and aqueous solutions.
The absolute-scale electronic energetics of liquid water and aqueous solutions, both in the bulk and at associated interfaces, are the central determiners of water-based chemistry. However, such information is generally experimentally inaccessible. Here we demonstrate that a refined implementation of the liquid microjet photoelectron spectroscopy (PES) technique can be adopted to address this. Implementing concepts from condensed matter physics, we establish novel all-liquid-phase vacuum and equilibrated solution-metal-electrode Fermi level referencing procedures. This enables the precise and accurate determination of previously elusive water solvent and solute vertical ionization energies, VIEs. Notably, this includes quantification of solute-induced perturbations of water's electronic energetics and VIE definition on an absolute and universal chemical potential scale. Defining and applying these procedures over a broad range of ionization energies, we accurately and respectively determine the VIE and oxidative stability of liquid water as 11.33 ± 0.03 eV and 6.60 ± 0.08 eV with respect to its liquid-vacuum-interface potential and Fermi level. Combining our referencing schemes, we accurately determine the work function of liquid water as 4.73 ± 0.09 eV. Further, applying our novel approach to a pair of exemplary aqueous solutions, we extract absolute VIEs of aqueous iodide anions, reaffirm the robustness of liquid water's electronic structure to high bulk salt concentrations (2 M sodium iodide), and quantify reference-level dependent reductions of water's VIE and a 0.48 ± 0.13 eV contraction of the solution's work function upon partial hydration of a known surfactant (25 mM tetrabutylammonium iodide). Our combined experimental accomplishments mark a major advance in our ability to quantify electronic-structure interactions and chemical reactivity in liquid water, which now explicitly extends to the measurement of absolute-scale bulk and interfacial solution energetics, including those of relevance to aqueous electrochemical processes.


Paravertebral block anesthesia for inguinal hernia repair.
Choice of anesthesia for inguinal hernia repair remains a controversial topic. Local anesthesia has been described in the literature as the optimal technique, however general and spinal anesthesia are commonly used in practice despite well-known complications and side effects. The regional technique of paravertebral block has been successfully used at our institution for the operative treatment of breast cancer. Its attributes are prolonged sensory block with minimization of postoperative pain, reduction of nausea and vomiting, shortened hospital stay, patient satisfaction, and rapid return to normal activities. These features are desirable in the practice of ambulatory hernia surgery; hence we initiated the use of thoracic/lumbar paravertebral block for that surgical procedure. Paravertebral block anesthesia was performed on 30 consecutive patients. Block placement took an average of 12.3 minutes, in six cases repeat injection at one or two spinal levels was required due to incomplete blockade. Paravertebral block achieved effective anesthesia in 28 of 30 cases; conversion to general anesthesia was performed for two failed blocks. We are reporting postoperative data on the 28 completed blocks. Supplementation of intravenous sedation or injection of local anesthesia successfully treated transient intraoperative pain in 10 cases. Epidural extension of anesthesia resulting in lower limb numbness and motor weakness delayed the discharge of two patients. There were no other complications of anesthesia and no cases of urinary retention. TIme to onset of pain averaged 15 hours, while duration of sensory block was 13 hours. Patients were prescribed a standing order of naproxen 500 mg B.I.D. for 4 days regardless of pain; supplemental oral narcotic use during the 48 hours following surgery averaged 3.5 tablets, with 6 patients not requiring any narcotic. ninety-six percent of patients scheduled for ambulatory surgery were discharged from the postanesthesia care unit, with an average stay of 2.5 hours. Employed patients returned to work on day 5.5 (range 3-10 days); patients who were not employed returned to regular activities in 5.8 days (range 1-14 days). Eighty-two percent of patients reported being "very satisfied" with the anesthetic technique.


Divergent regulation of acetylcholinesterase and butyrylcholinesterase in tissues of the rat.
Investigating the possibility that acetylcholinesterase (AchE) and butyrylcholinesterase (BuChE) are regulated in a coordinated manner, we have examined the natural variation in activity of these two enzymes in several tissues of adult male Sprague-Dawley, Fischer-344, and Wistar-Furth rats. Both enzymes varied greatly in mean activity among brain, diaphragm, atria, serum, superior cervical ganglia, and liver. In Sprague-Dawley rats there were also large individual variations with up to a fivefold range of AChE activities and up to a 100-fold range of BuChE activities in a given tissue. Individual variations in cholinesterase activities appeared to be smaller in the inbred Fischer-344 or Wistar-Furth rats. Experiments with internal standards of partially purified AChE and BuChE indicated that the individual variations probably reflected differences in the intrinsic content or specific activity of the tissue enzymes. Comparison of the AChE activities in different tissues of a given group of rats failed to reveal statistically significant correlations in any strain (i.e., the relative activity of any one tissue was no guide to the relative activity of any other tissue in the same rat). This result indicates that the regulation of AChE is tissue-specific. By contrast, BuChE activity showed highly significant correlations among the majority of the tissues examined in the Sprague-Dawley rats, implying that widely dispersed factors can affect the regulation of this enzyme. Body-wide regulation is not necessarily the rule, however, since only a single tissue pair in the inbred Fischer rats and none of the pairs in the Wistar-Furth rats showed significant correlations of BuChE activity. In general, AChE and BuChE activities were not correlated with each other to a statistically significant degree. We conclude that the control of these enzymes normally involves different mechanisms and is strongly affected by the genetic background of the sample population.


On the structural relations of malachite. I. The rosasite and ludwigite structure families.
The crystal structures of malachite Cu(2)(OH)(2)CO(3) and rosasite (Cu,Zn)(2)(OH)(2)CO(3), though not isotypic, are closely related. A previously proposed approach explaining this relation via a common hypothetical parent structure is elaborated upon on the basis of group-subgroup considerations, leading to the conclusion that the aristotype of malachite and rosasite should crystallize in the space group Pbam (No. 55). An ICSD database search for actual representatives of this aristotype leads to the interesting observation that the structure type of ludwigite (Mg,Fe)(2)FeO(2)BO(3), which is adopted by several natural and synthetic oxide orthoborates M(3)O(2)BO(3), is closely related to the proposed malachite-rosasite aristotype and thus to the malachite-rosasite family of hydroxide carbonates M(2)(OH)(2)CO(3) in general. Relations within both structure families and their analogies are summarized in a joint simplified Bärnighausen tree.


Thermosensitivity of the reverse transcription process as an inactivation mechanism of lentiviral vectors.
Lentiviral vectors are an important tool for gene transfer research and gene therapy purposes. However, the low stability of these vectors affects their production, storage, and efficacy in preclinical and clinical settings. In the present work the mechanism underlying the thermosensitivity of lentiviral vectors was evaluated. For lentiviral vectors pseudotyped with amphotropic and RDpro envelopes, the capacity to perform reverse transcription was lost rapidly at 37 degrees C, in high correlation with the loss of infectivity. The vector with RDpro envelope presented a higher level of stability than that with amphotropic envelope for both the reverse transcription process and viral infectivity. Reverse transcriptase enzyme inactivation and viral template RNA degradation were not implicated in the loss of the viral capacity to perform reverse transcription. Furthermore, early entry steps in the infection process do not determine the rate of viral inactivation, as the amount of viral RNA and p24 protein entering the cells decreased slowly for both vectors. Taken together, it can be concluded that the reverse transcription process is thermolabile and thus determines the rate of lentiviral inactivation. Strategies to stabilize the reverse transcription process should be pursued to improve the applicability of lentiviral vectors in gene therapy.


Lessons from an International Initiative to Set and Share Good Practice on Human Health in Environmental Impact Assessment.
Environmental Impact Assessment (EIA) is applied to infrastructure and other large projects. The European Union EIA Directive (2011/92/EU as amended by 2014/52/EU) requires EIAs to consider the effects that a project might have on human health. The International Association for Impact Assessment and the European Public Health Association prepared a reference paper on public health in EIA to enable the health sector to contribute to this international requirement. We present lessons from this joint action. We review literature on policy analysis, impact assessment and Health Impact Assessment (HIA). We use findings from this review and from the consultation on the reference paper to consider how population and human health should be defined; how the health sector can participate in the EIA process; the relationship between EIA and HIA; what counts as evidence; when an effect should be considered 'likely' and 'significant'; how changes in health should be reported; the risks from a business-as-usual coverage of human health in EIA; and finally competencies for conducting an assessment of human health. This article is relevant for health authorities seeking to ensure that infrastructure, and other aspects of development, are not deleterious to, but indeed improve, human health.


Culture in reduced levels of oxygen promotes clonogenic sympathoadrenal differentiation by isolated neural crest stem cells.
Isolated neural crest stem cells (NCSCs) differentiate to autonomic neurons in response to bone morphogenetic protein 2 (BMP2) in clonal cultures, but these neurons do not express sympathoadrenal (SA) lineage markers. Whether this reflects a developmental restriction in NCSCs or simply inappropriate culture conditions was not clear. We tested the growth and differentiation potential of NCSCs at approximately 5% O(2), which more closely approximates physiological oxygen levels. Eighty-three percent of p75(+)P(0-) cells isolated from embryonic day 14.5 sciatic nerve behaved as stem cells under these conditions, suggesting that this is a nearly pure population. Furthermore, addition of BMP2 plus forskolin in decreased oxygen cultures elicited differentiation of thousands of cells expressing tyrosine hydroxylase, dopamine-beta-hydroxylase, and the SA lineage marker SA-1 in nearly all colonies. Such cells also synthesized and released dopamine and norepinephrine. These data demonstrate that isolated mammalian NCSCs uniformly possess SA lineage capacity and further suggest that oxygen levels can influence cell fate. Parallel results indicating that reduced oxygen levels can also promote the survival, proliferation, and catecholaminergic differentiation of CNS stem cells (Studer et al., 2000) suggests that neural stem cells may exhibit a conserved response to reduced oxygen levels.


Molecular Insights on Post-chemotherapy Retinoblastoma by Microarray Gene Expression Analysis.
Management of Retinoblastoma (RB), a pediatric ocular cancer is limited by drug-resistance and drug-dosage related side effects during chemotherapy. Molecular de-regulation in post-chemotherapy RB tumors was investigated. cDNA microarray analysis of two post-chemotherapy and one pre-chemotherapy RB tumor tissues was performed, followed by Principle Component Analysis, Gene ontology, Pathway Enrichment analysis and Biological Analysis Network (BAN) modeling. The drug modulation role of two significantly up-regulated genes (p≤0.05) - Ect2 (Epithelial-cell-transforming-sequence-2), and PRAME (preferentially-expressed-Antigen-in-Melanoma) was assessed by qRT-PCR, immunohistochemistry and cell viability assays. Differential up-regulation of 1672 genes and down-regulation of 2538 genes was observed in RB tissues (relative to normal adult retina), while 1419 genes were commonly de-regulated between pre-chemotherapy and post- chemotherapy RB. Twenty one key gene ontology categories, pathways, biomarkers and phenotype groups harboring 250 differentially expressed genes were dys-regulated (EZH2, NCoR1, MYBL2, RB1, STAMN1, SYK, JAK1/2, STAT1/2, PLK2/4, BIRC5, LAMN1, Ect2, PRAME and ABCC4). Differential molecular expressions of PRAME and Ect2 in RB tumors with and without chemotherapy were analyzed. There was neither up- regulation of MRP1, nor any significant shift in chemotherapeutic IC50, in PRAME over-expressed versus non-transfected RB cells. Cell cycle regulatory genes were dys-regulated post-chemotherapy. Ect2 gene was expressed in response to chemotherapy-induced stress. PRAME does not contribute to drug resistance in RB, yet its nuclear localization and BAN information, points to its possible regulatory role in RB.


beta-Glucuronidase and hyperbilirubinaemia in breast-fed and formula-fed babies.
Breast milk or formula milk and serum samples from 34 breast-fed babies, 15 formula-fed babies, and their mothers were examined at 3 and 21 days of neonatal age. Infant faecal samples were obtained on day 21 only. At both ages serum total bilirubin concentrations were significantly higher in breast-fed than in formula-fed infants. Beta-glucuronidase activity in formula milk was negligible while that in human milk was considerable. Day 21 faecal beta-glucuronidase was higher in the breast-fed babies. In the breast-fed babies, serum bilirubin levels were related to concentrations of beta-glucuronidase in breast milk on both days 3 and 21 and to levels of faecal beta-glucuronidase on day 21. In 4 additional babies whose feeds were temporarily changed from breast milk to formula milk because of hyperbilirubinaemia there was a striking decrease in faecal beta-glucuronidase activity coincident with a fall in serum bilirubin. Breast milk beta-glucuronidase seems to be an important factor in the neonatal hyperbilirubinaemia of breast-fed babies.


Malabsorption in thyroid dysfunctions.
Studies of intestinal absorptive functions in 11 hyperthyroid and 14 hypothyroid patients showed that steatorrhoea was common in hyperthyroid patients and rare in hypothyroid patients. D-xylose malabsorption was present in both the groups of thyroid disorders being more common in hypothyroid one. Occasionally vitamin B12 malabsorption was found in 2 hypothyroid patients. Jejunal mucosal changes were almost unremarkable. Both fat and carbohydrate malabsorption in both groups of patients appeared to be casually related to the thyroid dysfunction.


ADAM10 is the physiologically relevant, constitutive alpha-secretase of the amyloid precursor protein in primary neurons.
The amyloid precursor protein (APP) undergoes constitutive shedding by a protease activity called alpha-secretase. This is considered an important mechanism preventing the generation of the Alzheimer's disease amyloid-beta peptide (Abeta). alpha-Secretase appears to be a metalloprotease of the ADAM family, but its identity remains to be established. Using a novel alpha-secretase-cleavage site-specific antibody, we found that RNAi-mediated knockdown of ADAM10, but surprisingly not of ADAM9 or 17, completely suppressed APP alpha-secretase cleavage in different cell lines and in primary murine neurons. Other proteases were not able to compensate for this loss of alpha-cleavage. This finding was further confirmed by mass-spectrometric detection of APP-cleavage fragments. Surprisingly, in different cell lines, the reduction of alpha-secretase cleavage was not paralleled by a corresponding increase in the Abeta-generating beta-secretase cleavage, revealing that both proteases do not always compete for APP as a substrate. Instead, our data suggest a novel pathway for APP processing, in which ADAM10 can partially compete with gamma-secretase for the cleavage of a C-terminal APP fragment generated by beta-secretase. We conclude that ADAM10 is the physiologically relevant, constitutive alpha-secretase of APP.


Teachers' emotional exhaustion during the COVID-19 pandemic: Levels, changes, and relations to pandemic-specific demands.
We aimed to identify the levels of and changes in emotional exhaustion experienced by teachers and principals during the COVID-19 pandemic and pandemic-related stressors and resources. In a German sample of 2157 teachers and 374 principals, we found high levels and an increase of emotional exhaustion. Results from multi-group structural equation modeling analyses indicated that health concerns and workload were positively and social support negatively related to emotional exhaustion. Additional analyses of an open response question confirmed that teachers and principals experienced their work during the COVID-19 pandemic as predominantly stressful. These results indicate the importance of supporting both teachers and principals in reducing their exhaustion to help students overcome the consequences of the COVID-19 pandemic.


Mechanical ventilator as a shared resource for the COVID-19 pandemic.
COVID-19, the causative agent of which is a new type of coronavirus called SARS-CoV-2, has caused the most severe pandemic in the last 100 years. The condition is mainly respiratory, and up to 5% of patients develop critical illness, a situation that has put enormous pressure on the health systems of affected countries. A high demand for care has mainly been observed in intensive care units and critical care resources, which is why the need to redistribute resources in critical medicine emerged, with an emphasis on distributive justice, which establishes the provision of care to the largest number of people and saving the largest number of lives. One principle lies in allocating resources to patients with higher life expectancy. Mechanical ventilator has been assumed to be an indivisible asset; however, simultaneous mechanical ventilation to more than one patient with COVID-19 is technically possible. Ventilator sharing is not without risks, but the principles of beneficence, non-maleficence and justice prevail. According to distributive justice, being a divisible resource, mechanical ventilator can be shared; however, we should ask ourselves if this action is ethically correct.


Haemophagocytosis in bone marrow aspirate--a review of the clinical course of 10 cases.
The clinical course of 10 cases where marrow aspirate showed features of haemophagocytosis was reviewed. Eight of these had a fulminant clinical course characterized by high fever, constitutional symptoms, wasting, hepatosplenomegaly with liver dysfunction, sometimes lymphadenopathy, progressive pancytopenia and coagulopathy, like that described as 'malignant histiocytosis' in the past. The remaining 2 cases did not have this classical clinical syndrome. Among the former 8 cases, 4 of them had high-grade lymphoma, 3 of whom were confirmed to be peripheral T cell lymphoma. Three of the remaining 4 had suspicious lymphomatous infiltrate on marrow trephine. In every case an extensive search for viral etiology by serology was negative. The 2 cases which did not have fulminant clinical feature were found to have lymphoma of the diffuse large cell and Ki-1 anaplastic type, respectively. A review of the literature reveal that most cases with haemophagocytic syndrome have a fulminant clinical course and are peripheral T cell lymphoma, which generally has a poor prognosis. In our study, the 8 cases with the classical haemophagocytic syndrome had a median survival of 24 days and a long-term survival of 37.5% at 28 months. Prompt initiation of chemotherapy is a life-saving measure and the only chance of achieving a long-term survival in patients with haemophagocytic syndrome if the underlying lymphoma can be diagnosed early.


Increasing reciprocal social interactions using collaborative art activities: An intervention for children with complex communication needs and their peers.
Purpose: Children with complex communication needs who require augmentative and alternative communication (AAC) strategies to communicate have difficulty participating in interactions with their peers. Integrating a collaborative learning framework with art activities may facilitate social interactions between children with complex communication needs and their peers.Method: A single-subject withdrawal design was utilised to examine the impact of collaborative art activities on reciprocal social interactions between 2 dyads of participants with complex communication needs and their same-age peers. Additional dependent variables included the roles of participants with complex communication needs in interactions (initiator or responder), AAC modalities (unaided, aided, natural speech), and engagement.Result:Tau-U calculations for Dyad 1 demonstrated 64% improvement from baseline, indicating moderate effects with the collaborative art activity (Intervention One) and 100% improvement, indicating strong effects following the activity enhanced with photography (Intervention Two). Dyad 2 demonstrated 100% improvement from baseline following Intervention One and 96% improvement following Intervention Two, indicating strong effects.Conclusion: Both collaborative art activities demonstrated potential for facilitating social interactions for children with complex communication needs and their peers.


Efficacious immune therapy in chronic myelogenous leukemia (CML) recognizes antigens that are expressed on CML progenitor cells.
Curative effects of graft-versus-leukemia-based therapies such as donor lymphocyte infusion (DLI) for chronic myelogenous leukemia (CML) may result from immunologic ablation of self-renewing CML progenitor cells. Patients who achieved durable remissions after DLI developed a significant B-cell lymphocytosis after treatment, which did not occur in patients who were unresponsive to DLI. In this study, we identified antigen targets of this B-cell response by probing two immunoproteomic platforms with plasma immunoglobulins from seven CML patients with clinically apparent graft-versus-leukemia responses after DLI. In total, 62 antigens elicited greater reactivity from post-DLI versus pre-DLI plasma. Microarray analysis revealed that >70% of the antigens were expressed in CML CD34(+) cells, suggesting that expression in malignant progenitor cells is a feature common to antibody targets of DLI. We confirmed elevated expression of three target antigens (RAB38, TBCE, and DUSP12) in CML that together consistently elicited antibody responses in 18 of 21 of an additional cohort of CML patients with therapeutic responses, but not in normal donors and rarely in non-CML patients. In summary, immunologic targets of curative DLI responses include multiple antigens on CML progenitor cells, identifying them as potential immunogens for vaccination and/or monitoring of immunotherapeutics designed to eliminate myeloid leukemia stem cells.


[A case of medial medullary infarction with persistent primitive hypoglossal artery].
A 66-year-old woman was admitted to our hospital because of vomiting, dizziness and vertigo. Neurological examination on admission revealed only upbeat nystagmus without cranial nerve symptoms, paresis, cerebellar signs or sensory disturbances. Magnetic resonance(MR) images demonstrated a new T 2 high intensity and T 1 iso-intensity signal lesion in the right upper medial medulla. This medial medullary infarction caused central vestibular dysfunction. MR angiography and digital subtraction angiography demonstrated a persistent primitive hypoglossal artery (PPHA) originating from the right internal carotid artery to the vertebrobasilar artery associated with the stenosis of the right internal carotid artery at the level of the cervical bifurcation. This is the first report of medullary infarction with persistent carotid-basilar anastomosis. We suspected this medullary infarction was caused by artery to artery embolism in the branch of the right vertebral artery through the PPHA distal originated from the stenosis of the right internal carotid artery.


Association of Induction Infliximab Levels With Clinical Response in Perianal Crohn's Disease.
The association of infliximab [IFX] trough levels with clinical and endoscopic outcomes in inflammatory bowel disease is well established. However, there is scarce data regarding the association of perianal fistula response with IFX. The aim of this study was to establish whether early induction infliximab levels and anti-infliximab antibodies [ATIs] are associated with perianal fistula response. Consecutive CD patients with perianal fistulae that were treated with IFX between 2008 and 2016 were included in the study. Response was defined as cessation or significant improvement of fistula drainage. Patients with unavailable IFX level or ATI measurements and/or missing clinical follow-up at Week 14 were excluded. A total of 36 patients with perianal fistulae were included; 25/36 [69.4%] responded to treatment by Week 14. The median induction IFX levels at Weeks 2, 6 and 14 in the responders group at Week 14 were higher compared with those of the non-responders group [20/5.6 µg/mL, P = 0.0001; 13.3/2.55 µg/mL P = 0.0001; 4.1/0.14 µg/mL, P = 0.01]. On multivariate analysis, IFX leve at Weeks 2 and 6 were significantly associated with fistula response at Weeks 14 and 30. IFX drug levels of 9.25 µg/mL at Week 2 and 7.25 µg/mL at Week 6 were the best predictors of fistula response. High IFX trough levels during induction are associated with favorable fistula response to anti-TNF treatment. If validated in a larger prospective study, our findings may help guide anti-TNF treatment in patients with perianal CD, and suggest serum level-guided treatment escalation in non-responders or prompt changing of biologic treatment in non-responders.


Enhanced prognosis of HCC patients undergoing radical treatments with tenofovir versus entecavir: A meta-analysis based on propensity score matching studies.
The selection of postoperative antiviral therapy for hepatocellular carcinoma (HCC) patients with HBV infection undergoing radical treatments (HPHR) is a topic of ongoing debate and controversy. The primary aim of this study was to compare the prognostic impact of selecting entecavir (ETV) or tenofovir disoproxil fumarate (TDF) as antiviral therapy options in HPHR. All the studies included in this analysis were implemented propensity score matching (PSM) methodology. Meta-analysis was performed using R statistical software (version 4.3.0). The primary outcome measures, overall survival (OS) and recurrence-free survival (RFS), were quantified using hazard ratios (HR) and 95% confidence intervals (CI). This study analyzed 13 studies involving 6961 patients (2394 in the TDF group and 4567 in the ETV group). We conducted a meta-analysis of 8 studies that included a total of 5289 patients using the PSM analysis method. In comparison to the ETV group, the TDF group demonstrated significantly better RFS (HR = 0.81; 95% CI, 0.70-0.93; p = 0.0034) and OS (HR = 0.61; 95% CI, 0.42-0.88; p = 0.0085). Furthermore, the disparity between the two drugs was particularly evident in the prognosis of patients undergoing hepatectomy. Regional disparities were observed, with mainland China studies favoring RFS benefits and Taiwan or Korea studies favoring OS benefits. In conclusion, TDF has demonstrated significant superiority over ETV in terms of RFS and OS outcomes for HPHR. The findings hold significant implications for informing clinical decision-making and guiding the selection of postoperative antiviral therapy drugs in HCC patients.


A multiplexed serum biomarker immunoassay panel discriminates clinical lung cancer patients from high-risk individuals found to be cancer-free by CT screening.
Clinical decision making in the setting of computed tomography (CT) screening could benefit from accessible biomarkers that help predict the level of lung cancer risk in high-risk individuals with indeterminate pulmonary nodules. To identify candidate serum biomarkers, we measured 70 cancer-related proteins by Luminex xMAP (Luminex Corporation) multiplexed immunoassays in a training set of sera from 56 patients with biopsy-proven primary non-small-cell lung cancer and 56 age-, sex-, and smoking-matched CT-screened controls. We identified a panel of 10 serum biomarkers-prolactin, transthyretin, thrombospondin-1, E-selectin, C-C motif chemokine 5, macrophage migration inhibitory factor, plasminogen activator inhibitor, receptor tyrosine-protein kinase, erbb-2, cytokeratin fragment 21.1, and serum amyloid A-that distinguished lung cancer patients from controls with an estimated balanced accuracy (average of sensitivity and specificity) of 76.0 ± 3.8% from 20-fold internal cross-validation. We then iteratively evaluated this model in an independent test and verification case/control studies confirming the initial classification performance of the panel. The classification performance of the 10-biomarker panel was also analytically validated using enzyme-linked immunosorbent assays in a second independent case/control population, further validating the robustness of the panel. The performance of this 10-biomarker panel-based model was 77.1% sensitivity/76.2% specificity in cross-validation in the expanded training set, 73.3% sensitivity/93.3% specificity (balanced accuracy 83.3%) in the blinded verification set with the best discriminative performance in stage I/II cases: 85% sensitivity (balanced accuracy 89.2%). Importantly, the rate of misclassification of CT-screened controls was not different in most control subgroups with or without airflow obstruction or emphysema or pulmonary nodules. These biomarkers have potential to aid in the early detection of lung cancer and more accurate interpretation of indeterminate pulmonary nodules detected by CT screening.


Serodiagnosis of multiple cancers using an extracellular protein kinase A autoantibody-based lateral flow platform.
Extracellular protein kinase A autoantibody (ECPKA-AutoAb) has been suggested as a universal cancer biomarker due to its higher amounts in serum of several types of cancer patients than that of normal individuals. Herein, we first developed a lateral flow immunoassay (LFIA) tool, using a sandwich format, toward ECPKA-AutoAb in human serum. For this format, 3G2 as a capture antibody was identified using hybridoma technique and a series of screenings where it showed superior capacity to recognize Enzo PKA catalytic subunit alpha (Cα), compared to other PKA antibodies and antigens. Using these components, we performed sandwich ELISA toward a mimic and real sample of ECPKA-AutoAb. As per the results, limit of detection (LOD) was found to be 135 ng/mL and ECPKA-AutoAb levels were higher in various cancer patients than in normal individuals like previous studies. Based on these results, we applied this sandwich format into LFIA tool and found that the LOD of the fabricated LFIA tool showed about 3.8 ng/mL using spiked PKA-Ab, which is significantly improved compared to the LOD of sandwich ELISA. Also, the developed LFIA tool demonstrated a remarkable ability to detect significant differences in ECPKA-AutoAb levels between normal and cancer patients within 15 min, showing a potential for point-of-care (PoC) detection. One interesting point is that our LFIA strip contains an additional conjugation pad II, named because of its position behind the conjugation pad, in which PKA Cα is dried, enabling a sandwich format.


The CREST Simulation Development Process: Training the Next Generation.
The challenges of training and assessing endourologic skill have driven the development of new training systems. The Center for Research in Education and Simulation Technologies (CREST) has developed a team and a methodology to facilitate this development process. Backwards design principles were applied. A panel of experts first defined desired clinical and educational outcomes. Outcomes were subsequently linked to learning objectives. Gross task deconstruction was performed, and the primary domain was classified as primarily involving decision-making, psychomotor skill, or communication. A more detailed cognitive task analysis was performed to elicit and prioritize relevant anatomy/tissues, metrics, and errors. Reference anatomy was created using a digital anatomist and clinician working off of a clinical data set. Three dimensional printing can facilitate this process. When possible, synthetic or virtual tissue behavior and textures were recreated using data derived from human tissue. Embedded sensors/markers and/or computer-based systems were used to facilitate the collection of objective metrics. A learning Verification and validation occurred throughout the engineering development process. Nine endourology-relevant training systems were created by CREST with this approach. Systems include basic laparoscopic skills (BLUS), vesicourethral anastomosis, pyeloplasty, cystoscopic procedures, stent placement, rigid and flexible ureteroscopy, GreenLight PVP (GL Sim), Percutaneous access with C-arm (CAT), Nephrolithotomy (NLM), and a vascular injury model. Mixed modalities have been used, including "smart" physical models, virtual reality, augmented reality, and video. Substantial validity evidence for training and assessment has been collected on systems. An open source manikin-based modular platform is under development by CREST with the Department of Defense that will unify these and other commercial task trainers through the common physiology engine, learning management system, standard data connectors, and standards. Using the CREST process has and will ensure that the systems we create meet the needs of training and assessing endourologic skills.


Consulting with the anxious patient, Part I: Complexities of the consultation process and problems of referral.
At least 20% of medical patients have anxiety disorders and are in need of treatment, but work with anxious patients is complicated by their distortions and often negative attitudes toward physicians and hospitals. Consultation is complicated by inconsistent referral patterns. Most anxious medical patients accept psychiatric consultation: only 15% to 20% have negative attitudes toward psychiatry. Guidelines for referral are presented.


[Autologous stem cell therapy with surgical myocardial revascularization - The Rostock University experience].
Stem cell therapy has gained great attraction in the treatment of acute and chronic myocardial diseases in recent years. The aim was to evaluate our experience in light of the middle term results of intramyocardial stem cell treatment with concomitant coronary artery bypass surgery (CABG) since 2001. After encouraging initial results of the Phase I (safety) trial with the first 15 patients, a prospective, controlled Phase II (efficacy) study was begun with 40 patients aiming to determine functional benefit of stem cell treatment using bone marrow derived CD 133+stem cell therapy with concomitant CABG in comparison to CABG only since 2003. Medium-term results of intervention were evaluated using patient's clinical findings, Holter monitoring, echocardiography, magnetic resonance imaging, computed tomography and myocardial scintigraphy. Statistical analyses were performed using unpaired t, Mann-Whitney U, ANOVA for repeated measurements and Chi-square tests. Left ventricular ejection fraction (LVEF) has increased significantly at 6th and 18th months follow-up in the first 15 patients who received therapy since 2001 (preoperative: 39.0+/- 8.7%; 6th month: 50.2+/- 8.5% and 18th month: 47.9+/- 6.0%; p=0.012). In the late group of patients, LVEF increased from 37.4+/- 8.4% to 47.1+/- 8.3% (p<0.001) whilst although an increase in LVEF has been observed in the CABG alone group (from 37.9+/- 10.3% to 41.3+/- 9.1%) the increase has not been statistically significant. Mortality occurred in 2 patients [1 patient from the early and 1 patient from the medium term follow-up] due to non-cardiac reasons. Myocardial calcification, lethal ventricular arrhythmia, and tumor formation have not been observed in any of the patients in the long-term follow up. Direct injection of bone marrow derived CD 133+ stem cells into the myocardium with concomitant CABG is safe. However, this treatment modality may only be applied as standard treatment after completion of the long- term detailed results of prospective, randomized multicenter trials.


[Analysis of thermodenaturation of Na,K-ATPase in the rat myocardial sarcolemma and possible role of damage of this enzyme in the pathogenesis of arrhythmias].
Activity of Na+, K+-ATPase and kinetics of the enzyme thermodenaturation were estimated in preparations of heavy sarcolemma isolated from rat myocardium. Emotional-painful stress (EPS) decreased the enzymatic activity by 20%, whereas the rate of the enzyme thermodenaturation was increased 2-3-fold. A thermodynamic analysis enabled to find a decrease in the energy of activation of the process after the stressory effects as well as alterations in entalpy and entropy under conditions of practically constant changes in free energy; this suggests the possible conformational transformations in the Na+, K+-ATPase molecules in EPS, resembling the state of denaturation. Lipid peroxidation was essential for the decrease in the enzyme thermostability. Activation of lipid peroxidation in the sarcolemma native membranes was accompanied by impairments typical for EPS. Preadministration of an antioxidant ionol before stress protected both the activity and thermostability of Na+, K+-ATPase. Adaptation to EPS by means of multiple short-term stressory actions prevented also the enzyme impairment. Mechanisms of the stress arrhythmogenic effect are discussed.


Prevention of drowning in home pools--lessons from Australia.
As an affluent and warm weather country, Australia faced an epidemic of drowning in children under five as the popularity of home pools increased in the late 1960s and early 1970s. In one state child drowning rose by 6.62 per 100,000 in the five years from 1968. Variation across jurisdictions in actions to regulate domestic pools has resulted in Australia representing an unfortunate natural experiment in interventions to prevent child drowning in pools. Mechanisms adopted by various authorities were analysed in detail to identify factors that contributed to success and failure in regulating domestic pools. Despite early identification of the problem and development of what were later demonstrated to be effective interventions, best practice solutions were not established in any jurisdiction until 2002, 25 years after identification of the problem. Key failures in intervention were found to be: failure to act in a timely way, flawed regulation, blockage of timely action and repeal of effective state regulation under political pressure, and blockage of effective standards. Lessons are identified as: inherent weaknesses in the standard setting process, failure of interventions to address major risk categories, failure to act before the hazard grew, failure to consider lesser interventions for tactical reasons, failure to pay attention to advocates, and the need for continuing effort. The paper argues that analysis of the identification of the problem, development of interventions and implementation of mechanisms of prevention illustrate where systemic changes are required and indicate lessons that should be applied in developing and implementing future interventions.


Biotic signals associated with benthic impacts of salmon farms from eDNA metabarcoding of sediments.
Environmental DNA (eDNA) metabarcoding can rapidly characterize the composition and diversity of benthic communities, thus it has high potential utility for routine assessments of benthic impacts of marine finfish farming. In this study, 126 sediment grab samples from 42 stations were collected at six salmon farms in British Columbia, Canada. Benthic community changes were assessed by both eDNA metabarcoding of metazoans and macrofaunal polychaete surveys. The latter was done by analysing 11,466 individuals using a combination of morphology-based taxonomy and DNA barcoding. Study objectives were to: (i) compare biotic signals associated with benthic impacts of salmon farming in the two data sources, and (ii) identify potential eDNA indicators to facilitate monitoring in Canada. Alpha diversity parameters were consistently reduced near fish cage edge and negatively correlated with pore-water sulphide concentration, with coefficients ranging from -0.62 to -0.48. Although Polychaeta are a common indicator group, the negative correlation with pore-water sulphide concentration was much stronger for Nematoda OTU richness (correlation coefficient: -0.86) than for Polychaeta (correlation coefficient: -0.38). Presence/absence of Capitella generally agreed well between the two methods despite that they differed in the volume of sediments sampled and the molecular marker used. Multiple approaches were used to identify OTUs related to organic enrichment statuses. We demonstrate that eDNA metabarcoding generates biotic signals that could be leveraged for environmental assessment of benthic impacts of fish farms in multiple ways: both alpha diversity and Nematoda OTU richness could be used to assess the spatial extent of impact, and OTUs related to organic enrichment could be used to develop local biotic indices.


Characterisation of a subtype of colorectal cancer combining features of the suppressor and mild mutator pathways.
10% of sporadic colorectal cancers are characterised by a low level of microsatellite instability (MSI-L). These are not thought to differ substantially from microsatelite-stable (MSS) cancers, but MSI-L and MSS cancers are distinguished clinicopathologically and in their spectrum of genetic alterations from cancers showing high level microsatellite instability (MSI-H). To study the distribution of molecular alterations in a series of colorectal cancers stratified by DNA microsatellite instability. A subset of an unselected series of colorectal cancers was grouped by the finding of DNA MSI at 0 loci (MSS) (n = 51), 1-2 loci (MSI-L) (n = 38) and 3-6 loci (MSI-H) (n = 25). The frequency of K-ras mutation, loss of heterozygosity (LOH) at 5q, 17p and 18q, and patterns of p53 and beta catenin immunohistochemistry was determined in the three groups. MSI-H cancers had a low frequency of K-ras mutation (7%), LOH on chromosomes 5q (0%), 17p (0%) and 18q (12.5%), and a normal pattern of immunostaining for p53 and beta catenin. MSI-L cancers differed from MSS cancers in terms of a higher frequency of K-ras mutation (54% v 27%) (p = 0.01) and lower frequency of 5q LOH (23% v 48%) (p = 0.047). Whereas aberrant beta catenin expression and 5q LOH were concordant (both present or both absent) in 57% of MSS cancers, concordance was observed in only 20% of MSI-L cancers (p = 0.01). MSI-L colorectal cancers are distinct from both MSI-H and MSS cancers. This subset combines features of the suppressor and mutator pathways, may be more dependent on K-ras than on the APC gene in the early stages of neoplastic evolution, and a proportion may be related histogenetically to the serrated (hyperplastic) polyp.


Psychometric development of the hypersexual behavior consequences scale.
Background and aims The past decade has seen an increased interest in understanding hypersexual behavior and its associated features. Beyond the obvious risks for sexually transmitted infections, there is a paucity of literature examining specific challenges encountered by hypersexual individuals. This study investigated and developed a new scale, the Hypersexual Behavior Consequences Scale (HBCS), to assess the various consequences reported among hypersexual patients. Methods Participants were drawn from a sample of patients recruited in a DSM-5 Field Trial for Hypersexual Disorder (HD). Participants completed the Hypersexual Behavior Inventory, a structured diagnostic interview to assess for psychopathology and HD, and self-report measures of personality, life satisfaction, and the initial item pool for the HBCS. Results Factor analysis reduced the HBCS items to a single factor solution which showed high internal consistency and stability over time. Higher HBCS scores were positively correlated with higher levels of emotional dysregulation, impulsivity, and stress proneness and lower levels of satisfaction with life and happiness. HBCS scores among the hypersexual patients were significantly higher than non-hypersexual patients. Conclusions The HBCS possesses good psychometric properties and appears to capture various consequences associated with the DSM-5 proposed criteria for HD. The HBCS can be used to aid clinicians and researchers in identifying consequences associated with hypersexual behavior. The HBCS may also prove a useful tool to guide treatment interventions aimed at reducing the negative impact of hypersexuality in patient populations.


Prospects for gene therapy in sports medicine.
For the orthopedic sports medicine physician soft tissue injuries often present the greatest clinical problems. Not only do many of the most frequently injured tissues, such as the cruciate ligaments and articular cartilage, have very limited capabilities for spontaneous repair, but they also respond poorly to surgical or nonsurgical intervention. In this article we try to define the role of growth factors in these conditions and to outline concepts for future treatment based upon modulation of the native repair response. We suggest that gene transfer could improve the management of such injuries, particularly when used as vehicles for the targeted delivery of growth factors. The concept of gene therapy in orthopedic sports medicine can be extended to include disorders that present as laxity or mechanical weakness of ligaments. We speculate that subtle genetic differences between individuals may account for those who appear to be injury prone. In these cases it is likely that genes encoding the structural macromolecules of the matrix are defective. Local gene supplementation in such cases could be useful in the future.


Absence of Rac1 and Rac3 GTPases in the nervous system hinders thymic, splenic and immune-competence development.
The nervous system influences organ development by direct innervation and the action of hormones. We recently showed that the specific absence of Rac1 in neurons (Rac1(N) ) in a Rac3-deficient (Rac3(KO) ) background causes motor behavioural defects, epilepsy, and premature mouse death around postnatal day 13. We report here that Rac1(N) /Rac3(KO) mice display a progressive loss of immune-competence. Comparative longitudinal analysis of lymphoid organs from control, single Rac1(N) or Rac3(KO) , and double Rac1(N) /Rac3(KO) mutant animals showed that thymus development is preserved up to postnatal day 9 in all animals, but is impaired in Rac1(N) /Rac3(KO) mice at later times. This is evidenced by a drastic reduction in thymic cell numbers. Cell numbers were also reduced in the spleen, leading to splenic tissue disarray. Organ involution occurs in spite of unaltered thymocyte and lymphocyte subset composition, and proper mature T-cell responses to polyclonal stimuli in vitro. Suboptimal thymus innervation by tau-positive neuronal terminals possibly explains the suboptimal thymic output and arrested thymic development, which is accompanied by higher apoptotic rates. Our results support a role for neuronal Rac1 and Rac3 in dictating proper lymphoid organ development, and suggest the existence of lymphoid-extrinsic mechanisms linking neural defects to the loss of immune-competence.


Melatonin inhibits expression of the inducible isoform of nitric oxide synthase in murine macrophages: role of inhibition of NFkappaB activation.
The role of melatonin as an immunomodulator is well established. Recent reports showed that melatonin exerts protective effects in septic and hemorrhagic shock and in inflammation. The expression of the inducible isoform of nitric oxide synthase (iNOS) makes an important contribution to the pathophysiology of shock and inflammation. We studied, in cultured murine macrophages, the role of melatonin in the regulation of the expression of iNOS and defined the mode of melatonin's action. Our results show that melatonin, at 1 microM-1 mM, decreased the production of nitrite/nitrate (the breakdown products of NO) as well as the production of 6-keto-prostaglandin F1alpha (the major stable breakdown product of prostacyclin) in macrophages stimulated with bacterial lipopolysaccharide (10 microg/ml). We observed that melatonin reduces iNOS steady-state mRNA levels and iNOS protein expression in the same concentration range (1 microM-1 mM). Melatonin, up to 10 mM, exerted only a slight direct inhibitory effect on iNOS activity. Using iNOS promoter-luciferase constructs, we found that melatonin inhibits iNOS promoter activation. Inhibition of iNOS expression was associated with inhibition of activation of the transcription factor nuclear factor kappa B (NFkappaB). We conclude that melatonin inhibits NO production in immunostimulated macrophages mainly by inhibiting the expression of iNOS. This is due to inhibition of iNOS transcription, in part through inhibition of NFkappaB activation. Inhibition of iNOS-derived NO production by melatonin may contribute to the anti-inflammatory effects of this pineal secretory product.


Probing the functional impact of sub-retinal prosthesis.
Retinal prostheses are promising tools for recovering visual functions in blind patients but, unfortunately, with still poor gains in visual acuity. Improving their resolution is thus a key challenge that warrants understanding its origin through appropriate animal models. Here, we provide a systematic comparison between visual and prosthetic activations of the rat primary visual cortex (V1). We established a precise V1 mapping as a functional benchmark to demonstrate that sub-retinal implants activate V1 at the appropriate position, scalable to a wide range of visual luminance, but with an aspect-ratio and an extent much larger than expected. Such distorted activation profile can be accounted for by the existence of two sources of diffusion, passive diffusion and activation of ganglion cells' axons en passant. Reverse-engineered electrical pulses based on impedance spectroscopy is the only solution we tested that decreases the extent and aspect-ratio, providing a promising solution for clinical applications.


Effects of a resistance training intervention on the strength-deficit of elite young soccer players.
The aim of this study was to examine the effects of a traditional resistance training scheme on the relative strength (RS), relative peak-force (RPF), strength deficit (SDef), and vertical jump and sprint abilities in elite young soccer players. Thirty-five under-20 soccer players from two professional clubs were assessed before and after a 4-week competitive period. One team performed 12 sessions of a structured resistance training program and the other maintained their regular soccer-specific training and competitive routines. Resistance training sessions consisted of half-squat and jump-squat exercises at intensities of 30-80% of the one-repetition maximum. Both teams performed pre- and post-measurements in the following order: (1) countermovement jump (CMJ), (2) 20-m sprint, and (3) half-squat one-repetition maximum to determine the RS, RPF, and SDef. A two-way analysis of variance was used to test for group x time interaction among variables. Effect sizes (ES) and 95% confidence intervals (CI) were also calculated. Group x time interactions were demonstrated for RS ([ES [95%CI] = 1.21 [0.57; 1.85], P = 0.001), RPF (ES [95%CI] = 1.18 [0.52; 1.80], P = 0.001), SDef (ES [95%CI] = 0.86 [0.01; 1.71], P = 0.04), and CMJ (ES [95%CI] = 0.64 [0.28; 0.99], P = 0.001); whereas a non-significant interaction was observed for 20-m sprint performance (ES [95%CI] = 0.02 [-0.32; 0.36], P = 0.85). Traditional strength-power oriented training resulted in improved maximum strength performance and CMJ ability but, paradoxically, increased the SDef. As a consequence, stronger athletes are not necessarily able to use greater percentages of their peak-force against relatively lighter loads.


Changed prognosis of breech-presenting low birthweight infants.
We have compared the mortality and morbidity of low birthweight breech-presenting singleton infants born in the Coventry Maternity Hospital during the years 1973 and 1974 with those born during 1979 and 1980. During this period changes in management have included an increase in the caesarean section rate and the continuing development of methods of intensive care for the newborn infants. The neonatal mortality rate has fallen but the increased survival rate is accounted for largely by the survival of handicapped infants. The value of routine caesarean section in the absence of other obstetric pathology for low birthweight breeches is unproven and may be deleterious to mother and baby.


An overview of patient satisfaction with psychiatric treatment.
Patient satisfaction with psychiatric treatment can strongly influence pursuit and use of mental health services as well as treatment compliance and treatment outcome. Although assessment of patient satisfaction is crucial to designing effective mental health programs, no standard methodology exists to measure satisfaction; thus it is difficult to compare findings from different satisfaction studies. The author examines the studies in four areas of satisfaction research: patient satisfaction with treatment, with participation in research, with participation as subjects in psychiatric teaching, and with involuntary commitment. He notes the variance between mental health professionals' expectations of patient satisfaction and the higher satisfaction that patients themselves report. He also discusses the need for more study of the subjective experience of patients who participate in research projects, teaching conferences, and observed psychotherapy and who undergo involuntary commitment.


Long-term Risk of Pancreatic Malignancy in Patients With Branch Duct Intraductal Papillary Mucinous Neoplasm in a Referral Center.
Little is known about the development of branch duct intraductal papillary mucinous neoplasms (BD-IPMNs). We evaluated long-term outcomes of a large cohort of patients with BD-IPMNs to determine risk of malignancy and define a subset of low-risk BD-IPMNs. We performed a retrospective analysis of data from 577 patients with suspected or presumed BD-IPMN under surveillance at the Massachusetts General Hospital. Patients underwent cross-sectional imaging analysis at 3 months or later after their initial diagnosis. The diagnosis of BD-IPMN was based on the presence of unilocular or multilocular cysts of the pancreas and a non-dilated main pancreatic duct (<5 mm). We collected demographic, clinical, and pathology data. Cysts were characterized at the time of diagnosis and during the follow-up period. Follow-up duration was time between initial cyst diagnosis and date of last visit or death for patients without development of pancreatic cancer, date of surgery for patients with histologically confirmed malignancy, or date of first discovery of malignancy by imaging analysis for patients with unresectable tumors or who underwent neoadjuvant treatment before surgery. The primary outcome was risk of malignancy, with a focus on patients followed for 5 years or more, compared with that of the US population, based on standardized incidence ratio. Of the 577 patients studied, 479 (83%) were asymptomatic at diagnosis and 363 (63%) underwent endoscopic ultrasound at least once. The median follow-up time was 82 months (range, 6-329 months) for the entire study cohort; 363 patients (63%) underwent surveillance for more than 5 years, and 121 (21%) for more than 10 years. Malignancies (high-grade dysplasia or invasive neoplasm) developed after 5 years in 20 of 363 patients (5.5%), and invasive cancer developed in 16 of 363 patients (4.4%). The standardized incidence ratio for patients with BD-IPMNs without worrisome features of malignancy at 5 years was 18.8 (95% confidence interval, 9.7-32.8; P < .001). One hundred and eight patients had cysts ≤1.5 cm for more than 5 years of follow-up; only 1 of these patients (0.9%) developed a distinct ductal adenocarcinoma. By contrast, among the 255 patients with cysts >1.5 cm, 19 (7.5%) developed malignancy (P = .01). In a retrospective analysis of patients with BD-IPMNs under surveillance, their overall risk of malignancy, almost 8%, lasted for 10 years or more, supporting continued surveillance after 5 years. Cysts that remain ≤1.5 cm for more than 5 years might be considered low-risk for progression to malignancy.


[The foundation of research ethics in community law].
The creation of the European Community by the Treaty of Rome in 1957 marked the beginning of the efforts to coordinate and harmonize national policies in many strategic sectors with high economic value, among them several aspects of scientific research. The European Union Law, formerly known as European Community law, now includes a range of ethical principles that apply to research projects developed with the financial support of the European Union. Which were the steps in the integration of the ethics of sciences in the context of the Union? This article aims to study first, what were, and what are the legal bases of the integration of the ethical dimension of researches in life sciences by the European Union and, secondly, the institutional organisation that has been set up in order to discuss the development of common ethical norms, especially bioethics one, and in order to apply these rules which respect national particularities. In this regard, we analyse the relevant legal texts providing a foundation for the creation of a European bio-law and we give an overview of the European institutions' activity in the field of bioethics by looking particularly the health research field.


Ghrelin, growth and obesity.
The objective of this paper is to review the current evidence of a novel gastric hormone ghrelin. Ghrelin is an endogenous ligand for the growth hormone secretagogue receptor (GHS-R) that potently stimulates growth hormone (GH) release. The discovery of ghrelin opens up a new regulatory system for the GH secretion. Surprisingly, recent studies in rodents suggest that peripherally or centrally administrated ghrelin, independent of GH, decreases fat oxidation and increases food intake and adiposity. In addition, plasma ghrelin levels are lower in obese human subjects. Ghrelin might participate in meal initiation and may signal to the hypothalamus when an increase in the metabolic efficiency is needed. However, the role of ghrelin in the regulation of body weight in humans is unknown. Whether ghrelin's somatotrophic effect surpasses its adipogenic effect in the prolonged administration determines its effect on energy balance. New interesting implications for ghrelin have recently been suggested. For instance, ghrelin's cardiovascular effects might have clinical relevance. Furthermore, ghrelin might be involved in the growth of some neoplasms. In conclusion, ghrelin, a new somatotrophic, orexigenic and adipogenic peptide hormone, links the regulatory systems for growth and energy balance.


MicroRNAs: more than a role in plant development?
SUMMARY Eukaryotic small RNA comprises several classes of 21-25 nucleotide non-coding RNA, of which microRNA (miRNA) has gained a great deal of attention because it is directly involved in controlling growth and development in plants. miRNAs are processed by the RNase III-like Dicer, although recent studies have implicated additional gene products in the step-wise maturation of miRNAs from their primary nuclear transcripts. They function as sequence-specific guides to trigger cleavage or translational repression of target mRNAs that have complementary sequences. Natural miRNA targets encode members of large families of transcription factors, which are collectively required for a number of developmental processes. In addition to developmental regulation, some miRNAs might be involved in specific physiological responses to several types of stresses, such as those induced by pathogen infections. Strikingly, the potyviruses, the largest group of plant RNA viruses, are able to interfere with miRNA-guided cleavage of multiple regulatory targets in plants, thus modulating gene expression of the host cell.


Does maternal smoking during pregnancy cause childhood overweight?
The objective of this study was to examine a possible association between maternal smoking in pregnancy and childhood overweight. From a population-based cohort of 5722 women from Trondheim, Bergen (Norway) and Uppsala (Sweden) enrolled in early pregnancy during 1986-92, a random sample of 482 women was selected for participation. They were followed up throughout pregnancy, and their children from birth until 5 years of age. Data on maternal smoking and diet, socio-economic determinants and breast feeding were recorded prospectively. During pregnancy and childhood, anthropometric measures were also recorded. Maternal smoking status was based on reported number of cigarettes smoked in week 17 of pregnancy. Child overweight was defined by body mass index (BMI) and sum of skinfold thickness (SFT) >or= 85th percentile at 5 years of age. Children of mothers who smoked in pregnancy had increased risk of overweight at 5 years of age (RR 2.5, 95% CI 1.5, 4.2 for BMI; and RR 1.8, 95% CI 1.1, 3.0 for SFT). Adjusting for maternal diet, breast feeding, maternal obesity and socio-economic status did not suggest confounding. However, adjustment for birthweight increased the observed risk. A linear increase in BMI and SFT was observed with increasing number of cigarettes smoked. In conclusion, smoking during pregnancy may be a risk factor for development of childhood overweight. This study may support the hypothesis of 'fetal origin of adult disease', but the risk of overweight associated with smoking during pregnancy was independent of intrauterine growth retardation, and may thus be attributed to specific effects of cigarette smoke.


Transition-state analysis of 2-O-acetyl-ADP-ribose hydrolysis by human macrodomain 1.
Macrodomains, including the human macrodomain 1 (MacroD1), are erasers of the post-translational modification of monoadenosinediphospho-ribosylation and hydrolytically deacetylate the sirtuin product O-acetyl-ADP-ribose (OAADPr). OAADPr has been reported to play a role in cell signaling based on oocyte microinjection studies, and macrodomains affect an array of cell processes including transcription and response to DNA damage. Here, we investigate human MacroD1 by transition-state (TS) analysis based on kinetic isotope effects (KIEs) from isotopically labeled OAADPr substrates. Competitive radiolabeled-isotope effects and mass spectrometry were used to obtain KIE data to yield intrinsic KIE values. Intrinsic KIEs were matched to a quantum chemical structure of the TS that includes the active site residues Asp184 and Asn174 and a structural water molecule. Transition-state analysis supports a concerted mechanism with an early TS involving simultaneous nucleophilic water attack and leaving group bond cleavage where the breaking C-O ester bond=1.60 Å and the C-O bond to the attacking water nucleophile=2.30 Å. The MacroD1 TS provides mechanistic understanding of the OAADPr esterase chemistry.


Fast and Efficient Generation of Isogenic Induced Pluripotent Stem Cell Lines Using Adenine Base Editing.
Isogenic induced pluripotent stem cell (iPSC) lines are currently mostly created by homology directed repair evoked by a double-strand break (DSB) generated by CRISPR-Cas9. However, this process is in general lengthy and inefficient. This problem can be overcome, specifically for correction or insertion of transition mutations, by using base editing (BE). BE does not require DSB formation, hence avoiding creation of genomic off-target breaks and insertions and deletions, and as it is highly efficient, it also does not require integration of selection cassettes in the genome to enrich for edited cells. BE has been successfully used in many cell types as well as in some in vivo settings to correct or insert mutations, but very few studies have reported generation of isogenic iPSC lines using BE. Here, we describe a simple and fast workflow to generate isogenic iPSCs efficiently with a compound heterozygous or a homozygous Wolfram syndrome 1 (WFS1) mutation using adenine BE, without the need to include a genomic selection cassette and without off-target modifications. We demonstrated that correctly base-edited clones can be generated by screening only five cell clones in less than a month, provided that the mutation is positioned in a correct place with regards to the protospacer adjacent motif sequence and no putative bystander bases exist.


The influence of cortisone on experimental viral infection. VII. Kinetics of interferon formation and its inhibition with hydrocortisone in relation to viral strain and virulence.
A comparative study was undertaken of the pathogenesis of infection of the allantoic sac of the chick embryo with three influenza viruses of differing virulence, and of the influence of hydrocortisone on the course of infection. Judged on the basis of earlier onset and greater degree of inflammatory response and diminished survival time of infected embryos, Mel. and Lee viruses were markedly more virulent than PR8, despite the earlier appearance of virus in PR8-infected embryos. Interferon appeared first and in greater quantity in the allantoic fluid of Lee-infected embryos and latest with PR8 infection. Thus, there was no correlation of avirulence and better interferon production with the viruses under study in the present system. Furthermore, evidence obtained suggested that Lee virus ("virulent") was most susceptible to interferon action, and also that viral synthesis in the chorioallantoic membrane with PR8 ("avirulent") persisted after the appearance of interferon. The injection of hydrocortisone within 2 hr of the initiation of infection delayed the synthesis of all three viruses; had no significant effect upon the inflammatory response; and transiently inhibited the synthesis of interferon, while prolonging the survival of Lee- and Mel.-infected embryos. Late administration of hydrocortisone suppresses both the inflammatory response and the production of interferon. Only in the case of Lee virus infection did hydrocortisone administration lead to augmentation of final yields of virus with the low infection multiplicity employed in the present experiments. It is postulated that Lee virus is a better inducer of interferon because its infectivity in vivo is more rapidly inactivated. As a consequence synthesis of Lee virus is more under the control of endogenous interferon than is the case with PR8 or Mel. virus. Therefore, inhibition of interferon synthesis with hydrocortisone has a greater influence on final yields of Lee virus.