Lack of IL-15 results in the suboptimal priming of CD4+ T cell response against an intracellular parasite.
IFN-gamma-producing CD4+ T cells, although important for protection against acute Toxoplasma gondii infection, can cause gut pathology, which may prove to be detrimental for host survival. Here we show that mice lacking IL-15 gene develop a down-regulated IFN-gamma-producing CD4+ T cell response against the parasite, which leads to a reduction in gut necrosis and increased level of survival against infection. Moreover, transfer of immune CD4+ T cells from WT to IL-15-/- mice reversed inhibition of gut pathology and caused mortality equivalent to levels of parental WT mice. Down-regulated CD4+ T cell response in the absence of IL-15, manifested as reduced antigen-specific proliferation, was due to defective priming of the T cell subset by dendritic cells (DCs) of these animals. When stimulated with antigen-pulsed DCs from WT mice, CD4+ T cells from IL-15-/- mice were primed optimally, and robust proliferation of these cells was observed. A defect in the DCs of knockout mice was further confirmed by their reduced ability to produce IL-12 upon stimulation with Toxoplasma lysate antigen. Addition of exogenous IL-15 to DC cultures from knockout mice led to increased IL-12 production by these cells and restored their ability to prime an optimal parasite-specific CD4+ T cell response. To our knowledge, this is the first demonstration of the role of IL-15 in the development of CD4+ T cell immunity against an intracellular pathogen. Furthermore, based on these observations, targeting of IL-15 should have a beneficial effect on individuals suffering from CD4+ T cell-mediated autoimmune diseases.


Chronic hypoxic pulmonary hypertension. Is thrombin involved?
Thrombin contracts vascular smooth muscle and stimulates its proliferation. Using a specific thrombin inhibitor, hirudin, we studied whether thrombin contributes to the pulmonary vasoconstriction and vascular proliferation that occurs in pulmonary hypertension. Hirudin was infused intravenously (0.2 mg/h/kg) by minipumps in nine rats during a 3-wk exposure to hypobaric hypoxia (HH). Vehicle (normal saline) was infused in eight hypoxic control (HC) and seven normoxic control (NC) rats. Sufficient hirudin delivery was confirmed by a failure of undiluted plasma from HH, but not from NC and HC, to clot in response to thrombin. When the plasma samples were diluted 1:10, the thrombin time was significantly prolonged in HH when compared with that in both NC and HC. Although hirudin slightly reduced mean pulmonary arterial pressure in open-chest rats, there was no significant difference between the hypoxic groups in total pulmonary resistance, right ventricle weight, morphologic remodeling of lung vessels, or the perfusion pressure-flow relationship in isolated lungs. Vasoconstrictor responses of isolated lungs to angiotensin II and acute hypoxic challenges were not affected by hirudin treatment. We conclude that hirudin, in a dose sufficient to reduce thrombin's catalytic effect on fibrinogen, does not significantly prevent the development of chronic hypoxic pulmonary hypertension.


Navigating tuberculosis drug discovery with target-based screening.
Mycobacterium tuberculosis kills more people than any other bacterial pathogen. New drugs are required to shorten the treatment time and provide a viable therapy for drug-resistant and latent forms of tuberculosis. The tuberculosis field has advanced considerably since the publication of the M. tuberculosis genome sequence. Today, researchers can build a high definition map of the pathogen's traits and behavior and select individual targets for chemical disruption. This review examines the discovery of current clinical and candidate tuberculosis drugs. It outlines recent developments in the selection of molecular targets for the discovery of new anti-mycobacterial agents. It appraises techniques that incorporate target knowledge into the screening protocol. These techniques include in silico, in vitro enzyme-based, differential antisense sensitivity and gene expression screening systems. The review also looks ahead to further techniques that may be applied in tuberculosis drug discovery. The adoption of an 'either/or' approach to targeted or random tuberculosis drug screening is not expected. The historical success of random screening in providing the tuberculosis drugs currently in clinical use is likely to ensure that non-targeted protocols retain an important role in drug screening. However, a number of M. tuberculosis inhibitors in lead optimization and preclinical development have been discovered using targeted methods. Realization of the first clinically-approved tuberculosis drugs derived from targeted screening and continued refinements in targeted screening technologies are likely to increase the adoption of targeted approaches in the future.


Strain engineering of phonon thermal transport properties in monolayer 2H-MoTe2.
The effect of strain on the phonon properties such as phonon group velocity, phonon anharmonicity, phonon lifetime, and lattice thermal conductivity of monolayer 2H-MoTe2 is studied by solving the Boltzmann transport equation based on first principles calculations. The phonon thermal transport properties of the unstrained monolayer 2H-MoTe2 are compared to those of the strained case under different biaxial tensile strains. One of the common features of two-dimensional materials is the quadratic nature near the Γ point of the out-of-plane phonon flexural mode that disappears by applying tensile strain. We find that the lattice thermal conductivity of the monolayer 2H-MoTe2 is very sensitive to strain, and the lattice thermal conductivity is reduced by approximately 2.5 times by applying 8% biaxial tensile strain due to the reduction in phonon group velocities and phonon lifetime. We also analyze how the contribution of each mode to lattice thermal conductivity changes with tensile strain. These results highlight that tensile strain is a key parameter in engineering phonon thermal transport properties in monolayer 2H-MoTe2.


Reduction of ring artifacts in CBCT: detection and correction of pixel gain variations in flat panel detectors.
In using flat panel detectors (FPD) for cone beam computed tomography (CBCT), pixel gain variations may lead to structured nonuniformities in projections and ring artifacts in CBCT images. Such gain variations can be caused by change in detector entrance exposure levels or beam hardening, and they are not accounted by conventional flat field correction methods. In this work, the authors presented a method to identify isolated pixel clusters that exhibit gain variations and proposed a pixel gain correction (PGC) method to suppress both beam hardening and exposure level dependent gain variations. To modulate both beam spectrum and entrance exposure, flood field FPD projections were acquired using beam filters with varying thicknesses. "Ideal" pixel values were estimated by performing polynomial fits in both raw and flat field corrected projections. Residuals were calculated by taking the difference between measured and ideal pixel values to identify clustered image and FPD artifacts in flat field corrected and raw images, respectively. To correct clustered image artifacts, the ratio of ideal to measured pixel values in filtered images were utilized as pixel-specific gain correction factors, referred as PGC method, and they were tabulated as a function of pixel value in a look-up table. 0.035% of detector pixels lead to clustered image artifacts in flat field corrected projections, where 80% of these pixels were traced back and linked to artifacts in the FPD. The performance of PGC method was tested in variety of imaging conditions and phantoms. The PGC method reduced clustered image artifacts and fixed pattern noise in projections, and ring artifacts in CBCT images. Clustered projection image artifacts that lead to ring artifacts in CBCT can be better identified with our artifact detection approach. When compared to the conventional flat field correction method, the proposed PGC method enables characterization of nonlinear pixel gain variations as a function of change in x-ray spectrum and intensity. Hence, it can better suppress image artifacts due to beam hardening as well as artifacts that arise from detector entrance exposure variation.


Is epilepsy surgery on both hemispheres effective?
Patients with bilateral independent seizure foci are poor candidates for conventional resective epilepsy surgery. The authors have therefore used minimally invasive procedures to treat such patients. In this paper, the result of a large series of patients treated by this approach is examined to determine the effectiveness of this approach. The series had 61 patients. The range of follow-up was 15-90 months. The mean follow-up was 41.5 months with a median of 37 months. Patients' ages at the time of surgery ranged from 3 to 54 years, with a mean of 11.3 and median of 6. Male to female ratio was 39/22. Fifty patients had complex partial seizures, 3 had Lennox-Gastaut syndrome, 4 had myotonic seizures, 2 had infantile spasm, and 2 had myoclonic seizures. Preoperative evaluation included: video electroencephalogram (EEG) monitoring using scalp electrodes, neuropsychological evaluation, magnetic resonance scans, positron emission topography and/or single photon emission computed tomography scans, magnetoencephalogram, Wada test and video EEG recording using subdural electrodes. Multiple subpial transection (MST) was the principle procedure. This procedure was supplemented (17 patients) with minimal cortical resection when intraoperative electroencephalogram indicated that an area failed to respond to MST. When an additional epileptogenic focus was present in the amygdala-hippocampus complex (5 patients), it was treated with stereotactic amygdala-hippocampotomy. The eloquent cortex was treated in 51 patients. The number of lobes treated was 2 in 5 patients, 3 in 5 patients, 4 in 10 patients, 5 in 2 patients, 6 in 38 patients and 8 in 1. Seizure outcome based on Engel's modified classification was as follows: 32 patients (52.45%) were class I, 5 (8.2%) were class II, 15 (24.59%) were class III, 3 (4.9%) were class IV and 6 (9.83%) were class V. There was no statistical difference between those who were operated on of the first half of the series and those who were operated on of the second half of the series (p = 0.1636). Similarly, there was no statistical difference between this series and two large series in which MST had been performed on one hemisphere (p values of 0.6863 and 0.7337). There was no statistical difference between those who had MST alone and those who had MST plus minimal cortical resection (p = 0.1698). There was no permanent neurological complication in this series. Patients with intractable epilepsy with independent seizure foci in both hemispheres can be safely treated with the approach described in this paper, and seizure control achieved by this approach is fairly satisfactory and similar to that reported in patients with surgery on one hemisphere.


Solitary pulmonary metastasis presenting 20 years after primary resection of Wilms tumor.
Wilms tumor is the most common renal malignancy of childhood. Relapse occurs most often within 4 years of initial diagnosis, and the most common site of metastasis is the lung. We describe a 22-year-old man who presented with hemoptysis and a solitary pulmonary lesion 20 years after primary resection of Wilms tumor. Computed tomography of the chest showed an indeterminate pulmonary mass of heterogeneous attenuation with no other intrathoracic abnormalities. Surgical resection revealed a solitary pulmonary metastasis from Wilms tumor. Further evaluation yielded no evidence of extrathoracic metastases. This case shows that late relapse in the form of a solitary pulmonary mass can occur in patients with Wilms tumor.


Impact of Thermal Processing on the Selected Biological Activities of Ginger Rhizome-A Review.
Ginger (Zingiber officinale Rosc.) is a spice, medicinal and cosmetic plant that has been known for centuries. It can be used in dried, fresh, marinated or candied form, and is also an essential ingredient in well-known curry blends. Ginger rhizomes are often freeze-dried as the first step in the preparation of the raw material. Many studies have proved that the composition and biological activity of ginger changes due to thermal processing. Therefore, the aim of the review was to summarize the scientific results on the impact of traditional and unconventional methods of the heat treatment of ginger rhizomes and their influence on the antioxidant and other selected biological activities of the plant. The review of the available scientific data is inconclusive, and it is hard to state unequivocally whether the thermal treatment of the raw material increases or decreases biological activity. Based on the presented literature review, it can be concluded that traditional cooking and microwave processing in general decrease the antioxidant activity of the ginger rhizome, whereas frying, autoclaving, blanching or traditional drying in the sun mostly lead to a significant increase in ginger activity. Interesting data were presented in the works describing the freeze-drying process during which the antioxidant potential of ginger increased.


Amniotic membrane wrapping: an alternative method to the splenorrhaphy in the injured spleen.
Human amnion, which has been used experimentally and clinically as an ideal bioprosthesis, was used in the present study to wrap the experimentally injured spleens and the technique was compared to splenorrhaphy. Standard splenic injuries were made on 30 male albino rats. 15 of the spleens were repaired by splenorrhaphy and the other 15 were wrapped with sterile human amniotic membrane. Isotope uptakes of the spleens calculated at the 30th postoperative day were similar in both groups (p less than 0.05). But the histopathological examination revealed a better healing in the amnion group with better preservation of the splenic architecture. It was concluded that in heavily injured spleens, amnion wrapping may be an effective alternative to splenorrhaphy.


Spontaneous recovery of underlying bony erosion following surgical removal of craniofacial dermoid cysts in periatric patients: A prospective study.
This study aimed to evaluate the spontaneous recovery of bone deformity after surgical excision of craniofacial dermoid cysts in pediatrics. Pediatric patients who underwent excision of a dermoid cyst were included in the study. A prospective analysis was conducted to evaluate the amount of bone recovery by comparing the depth of bony concavity in the preoperative and postoperative (6 months) ultrasonography. In 145 of 187 patients with preoperative imaging available, the mean size of dermoid cysts was 1.4 cm3 (range, 0.1 to 9.5), and 41.4% (60/145 cases) showed cranial bone depression. In the comparison of preoperative and postoperative ultrasonography of 30 patients, the mean depth of bony cavity decreased significantly from 4.0 to 0.9 mm (p<0.001) after a mean of 6.7 months postoperatively. There was 13.3% (4/30) of mild (≤2.0 mm), 40.0% (12/30) of moderate (>2.0 to ≤4.0 mm), and 46.7% (14/30) of severe (>4.0 mm) depression, and the concavity depth significantly decreased in all groups (p = 0.028, mild; p<0.001, moderate; p<0.001 severe). Within the limitations of the study it seems that significant recovery of cranial bone depression does take place within 6 months after excision of craniofacial dermoid cysts in pediatric patients, saving the need for immediate reconstruction.


Amorphous High-Surface-Area Aluminum Hydroxide-Bicarbonates for Highly Efficient Methyl Orange Removal from Water.
Amorphous high-surface-area aluminum hydroxide-bicarbonates were synthesized starting from AlCl3, base, and bicarbonate in water. Composites with a chemical formulas of [Al13O4(μ-OH)24(H2O)6.5(OH)5.5](HCO3)1.5 (I-NaOH) and [Al13O4(μ-OH)24(H2O)6(OH)6](HCO3) (I-NH3) were obtained by the use of NaOH/NaHCO3 and NH3/NH4HCO3 as base/bicarbonate, respectively. The surface area of the composites was highly dependent on the pH level of the synthetic solution, and composites with high surface areas (ca. 200 m2 g-1) were obtained around pH 7-8. Pore-size distributions determined from the N2 adsorption isotherms showed that I-NH3 and I-NaOH possess mainly large (pore radius rp > 3 nm) and small (rp < 3 nm) pores, respectively, despite similar surface areas. While SEM images showed that both I-NH3 and I-NaOH were aggregates of nanoparticles, the particles were more fused in I-NaOH, which is in line with the existence of small pores and the use of a stronger base (NaOH), which would facilitate the dehydration condensation reaction. The composites were applied as adsorbents to remove methyl orange (MO) from water. The time course of MO adsorption was readily fitted with a pseudo-second-order model, and over 90% MO removal was attained within 10 min with I-NH3, while I-NaOH showed much less MO removal (26%). The MO adsorption isotherm of I-NH3 was reproduced with a Langmuir model with an adsorption capacity of 154 mg g-1. Notably, the aluminum hydroxide-bicarbonates could not absorb methylene blue, which is a cationic dye, while anions (MO and PO43-) were readily absorbed. Solid-state 27Al MAS NMR spectra showed that the concentration of 5-coordinated aluminum species, which may serve as guest binding sites, was higher for I-NH3. These results show that electrostatic interaction between anionic MO and coordinatively unsaturated 5-coordinated cationic aluminum species and the large external surface area of I-NH3 contribute to the highly efficient MO adsorption.


Haemoglobinopathies and glucose-6-phosphate dehydrogenase deficiency in a Scandinavian perspective.
Haemoglobinopathies (mainly thalassaemia and sickle-cell anaemia syndromes) and glucose-6-phosphate dehydrogenase deficiency (G6PD) are globally among the most prevalent single-genomic diseases. About 3% of the world's population are heterozygotic for beta-thalassaemia and about 1-2% for sickle-cell anaemia, and it is estimated that more than 400 million people are affected by G6PD deficiency worldwide. The disorders are most prevalent in the Mediterranean area, in Asia and Africa. The Scandinavian countries, among others, have seen a boom in immigration during the past 20 years, and therefore migration makes haemoglobinopathies as well as G6PD deficiency increasingly more important from a differential diagnostic perspective in most countries. The purpose of the present special issue of the Journal is to summarize current epidemiological data and elucidate trends and practices in the laboratory diagnosis of these disorders.


Halogenated peptides as internal standards (H-PINS): introduction of an MS-based internal standard set for liquid chromatography-mass spectrometry.
As the application for quantitative proteomics in the life sciences has grown in recent years, so has the need for more robust and generally applicable methods for quality control and calibration. The reliability of quantitative proteomics is tightly linked to the reproducibility and stability of the analytical platforms, which are typically multicomponent (e.g. sample preparation, multistep separations, and mass spectrometry) with individual components contributing unequally to the overall system reproducibility. Variations in quantitative accuracy are thus inevitable, and quality control and calibration become essential for the assessment of the quality of the analyses themselves. Toward this end, the use of internal standards cannot only assist in the detection and removal of outlier data acquired by an irreproducible system (quality control) but can also be used for detection of changes in instruments for their subsequent performance and calibration. Here we introduce a set of halogenated peptides as internal standards. The peptides are custom designed to have properties suitable for various quality control assessments, data calibration, and normalization processes. The unique isotope distribution of halogenated peptides makes their mass spectral detection easy and unambiguous when spiked into complex peptide mixtures. In addition, they were designed to elute sequentially over an entire aqueous to organic LC gradient and to have m/z values within the commonly scanned mass range (300-1800 Da). In a series of experiments in which these peptides were spiked into an enriched N-glycosite peptide fraction (i.e. from formerly N-glycosylated intact proteins in their deglycosylated form) isolated from human plasma, we show the utility and performance of these halogenated peptides for sample preparation and LC injection quality control as well as for retention time and mass calibration. Further use of the peptides for signal intensity normalization and retention time synchronization for selected reaction monitoring experiments is also demonstrated.


Anthelminthic efficacy of aqueous extract of Acanthus montanus leaf against strongylid nematodes of small ruminants.
The anthelminthic efficacy of the crude aqueous extract of Acanthus montanus (Nees) T. Anders (Acanthaceae) against strongylid nematodes of small ruminants was investigated using the in-vitro egg hatch and larval growth inhibition assays. Faecal samples collected per rectum from sheep and goats were subjected to parasitological examination using the McMaster counting technique with a yield of 700 eggs per gram (E.P.G.) of faeces. Crude aqueous leaf extract of Acanthus montanus was extracted using cold water extraction with a yield of 13.01% w/w. Egg hatch assay revealed a 91.75% reduction in egg hatch at concentration of 25 mg/ml of the extract. The extract had a 100% inhibition, at 200mg/ml concentration which was equivalent to the activity of 3.125 mg/ml albendazole. The distilled water control however, showed a 0% inhibition. Larval growth inhibition assay results showed the extract on Day 1 at 25 mg/ml and 200mg/ml concentrations yielded 67.02% and 85.26% larval inhibition. On Day 2, 100% inhibition was obtained on all concentrations of the extract except for 25 mg/ml that yielded 88.30%. Albendazole however, had a 100% larval inhibition for all concentrations on Day 2. A 100% larval inhibition was recorded on Day 3 from the 25 mg/ml concentrations. The mean percentage larval inhibition of the extract at 200mg/ml (92.63%) was comparable to the standard anthelminthic (albendazole) at 12.50mg/ml (92.28%). These findings showed that there is a pharmacological basis for the folkloric claim of the anthelminthic effect of Acanthus montanus.


Broadening the functionality of a J-protein/Hsp70 molecular chaperone system.
By binding to a multitude of polypeptide substrates, Hsp70-based molecular chaperone systems perform a range of cellular functions. All J-protein co-chaperones play the essential role, via action of their J-domains, of stimulating the ATPase activity of Hsp70, thereby stabilizing its interaction with substrate. In addition, J-proteins drive the functional diversity of Hsp70 chaperone systems through action of regions outside their J-domains. Targeting to specific locations within a cellular compartment and binding of specific substrates for delivery to Hsp70 have been identified as modes of J-protein specialization. To better understand J-protein specialization, we concentrated on Saccharomyces cerevisiae SIS1, which encodes an essential J-protein of the cytosol/nucleus. We selected suppressors that allowed cells lacking SIS1 to form colonies. Substitutions changing single residues in Ydj1, a J-protein, which, like Sis1, partners with Hsp70 Ssa1, were isolated. These gain-of-function substitutions were located at the end of the J-domain, suggesting that suppression was connected to interaction with its partner Hsp70, rather than substrate binding or subcellular localization. Reasoning that, if YDJ1 suppressors affect Ssa1 function, substitutions in Hsp70 itself might also be able to overcome the cellular requirement for Sis1, we carried out a selection for SSA1 suppressor mutations. Suppressing substitutions were isolated that altered sites in Ssa1 affecting the cycle of substrate interaction. Together, our results point to a third, additional means by which J-proteins can drive Hsp70's ability to function in a wide range of cellular processes-modulating the Hsp70-substrate interaction cycle.


[Changes in oxygen extraction rate after craniocerebral injury].
Changes of oxygen metabolism were observed in 8 experimental dogs and in 9 patients with head injury. Each dog harbored a bag in the epidural space, and was injected normal saline. The ICP was raised to 13.3 kPa and to 20.0 kPa respectively. Arterial oxygenation declined and mixed venous oxygenation went up when the ICP was raised to the given level and maintained for 30 minutes. The oxygen extraction rate decreased from 25.00 +/- 5.40% to 21.50 +/- 4.60% (P less than 0.01) at the ICP level of 20 kPa. Tissue hypoxia was observed in all patients. Although PaO was higher than inspiring air when patients inhaled 50% oxygen. Tissue hypoxia was not improved. The results suggest that the patients head injury is complicated by not only hypoxemia but decrease of oxygen extraction in tissue and organs. We consider that the alteration is responsible for multiple organs failure injury.


Comparative real-time effects on platelet adhesion and aggregation under flowing conditions of in vivo aspirin, heparin, and monoclonal antibody fragment against glycoprotein IIb-IIIa.
A real-time in vitro system of human platelet thrombosis under arterylike flowing conditions similar to those produced in vivo by angioplasty would be useful for the evaluation of potential antiarterial thrombotic agents in association with in vivo trials. Aspirin, heparin, and the chimeric monoclonal antibody antigen-binding fragment 7E3 (c7E3 Fab) directed against platelet glycoprotein (GP) IIb-IIIa have been used in attempts to delay or prevent thrombotic reocclusion of coronary arteries after angioplasty. We compared the effects of these agents administered in vivo on GPIb-mediated platelet adhesion to von Willebrand factor (vWF)/collagen type I (as in atherosclerotic subendothelium) and on subsequent GPIIb-IIIa-fibrinogen/vWF-mediated platelet aggregation under flowing conditions analogous to those in constricted coronary arteries. Citrated whole blood containing mepacrine-labeled platelets from patients and healthy donors was perfused for 1 minute at an abnormally elevated shear rate of 1500 seconds-1 (arterial wall shear stress of 50 to 60 dynes/cm2) at 37 degrees C over collagen I/vWF. The number of adherent fluorescent platelets was quantified every 15 seconds with a low-light-level video camera and epifluorescent microscopy. After 5 healthy donors had ingested 975 mg aspirin, platelet adhesion was unaffected in the aspirin-treated blood compared with the control blood in all experiments (10 of 10), and subsequent aggregation was unchanged in most runs (8 of 10). The blood of 3 aspirin-treated patients undergoing angioplasty was analyzed before and after a 12,000-U heparin injection and 2 minutes, 2 hours, and 24 hours after infusion of 0.25 mg/kg of c7E3 Fab. In these patients, the bolus of heparin did not inhibit either platelet adhesion to collagen I/vWF or subsequent aggregation. In contrast, there was > 50% inhibition of platelet aggregation 2 minutes after the infusion of c7E3 Fab in all 3 patients, and inhibition persisted in 2 of the 3 patients at 2 hours and 24 hours after c7E3 Fab. In contrast to aspirin or heparin, the in vivo injection of c7E3 Fab considerably reduces platelet aggregate formation mediated by the binding of fibrinogen, vWF, or some other ligand to platelet GPIIb-IIIa under conditions of abnormally increased shear stress analogous to those in narrowed coronary arteries. Platelet adherence to collagen I/vWF is not affected. This study describes an in vitro model of arterial injury (similar to angioplasty) that uses human blood to compare directly, in real time, the precise relative effects of aspirin, heparin, and c7E3 Fab on platelet adhesion and subsequent aggregation.


Observer Performance for Detection of Pulmonary Nodules at Chest CT over a Large Range of Radiation Dose Levels.
Background There is a wide variation in radiation dose levels that can be used with chest CT in order to detect indeterminate pulmonary nodules. Purpose To compare the performance of lower-radiation-dose chest CT with that of routine dose in the detection of indeterminate pulmonary nodules 5 mm or greater. Materials and Methods In this retrospective study, CT projection data from 83 routine-dose chest CT examinations performed in 83 patients (120 kV, 70 quality reference mAs [QRM]) were collected between November 2013 and April 2014. Reference indeterminate pulmonary nodules were identified by two nonreader thoracic radiologists. By using validated noise insertion, five lower-dose data sets were reconstructed with filtered back projection (FBP) or iterative reconstruction (IR; 30 QRM with FBP, 10 QRM with IR, 5 QRM with FBP, 5 QRM with IR, and 2.5 QRM with IR). Three thoracic radiologists circled pulmonary nodules, rating confidence that the nodule was a 5-mm-or-greater indeterminate pulmonary nodule, and graded image quality. Analysis was performed on a per-nodule basis by using jackknife alternative free-response receiver operating characteristic figure of merit (FOM) and noninferiority limit of -0.10. Results There were 66 indeterminate pulmonary nodules (mean size, 8.6 mm ± 3.4 [standard deviation]; 21 part-solid nodules) in 42 patients (mean age, 51 years ± 17; 21 men and 21 women). Compared with the FOM for routine-dose CT (size-specific dose estimate, 6.5 mGy ± 1.8; FOM, 0.86 [95% confidence interval: 0.80, 0.91]), FOM was noninferior for all lower-dose configurations except for 2.5 QRM with IR. The sensitivity for subsolid nodules at 70 QRM was 60% (range, 48%-72%) and was significantly worse at a dose of 5 QRM and lower, whether or not IR was used (P < .05). Diagnostic image quality decreased with decreasing dose (P < .001) and was better with IR at 5 QRM (P < .05). Conclusion CT images reconstructed at dose levels down to 10 quality reference mAs (size-specific dose estimate, 0.9 mGy) had noninferior performance compared with routine dose in depicting pulmonary nodules. Iterative reconstruction improved subjective image quality but not performance at low dose levels. © RSNA, 2020 Online supplemental material is available for this article. See also the editorial by White and Kazerooni in this issue.


Adaptive response in embryogenesis: IV. Protective and detrimental bystander effects induced by X radiation in cultured limb bud cells of fetal mice.
The radioadaptive response and the bystander effect represent important phenomena in radiobiology that have an impact on novel biological response mechanisms and risk estimates. Micromass cultures of limb bud cells provide an in vitro cellular maturation system in which the progression of cell proliferation and differentiation parallels that in vivo. This paper presents for the first time evidence for the correlation and interaction in a micromass culture system between the radioadaptive response and the bystander effect. A radioadaptive response was induced in limb bud cells of embryonic day 11 ICR mice. Conditioning irradiation of the embryonic day 11 cells with 0.3 Gy resulted in a significant protective effect against the occurrence of apoptosis, inhibition of cell proliferation, and differentiation induced by a challenging dose of 5 Gy given the next day. Both protective and detrimental bystander effects were observed; namely, irradiating 50% of the embryonic day 11 cells with 0.3 Gy led to a successful induction of the protective effect, and irradiating 70% of the embryonic day 12 cells with 5 Gy produced a detrimental effect comparable to that seen when all the cells were irradiated. Further, the bystander effect was markedly decreased by pretreatment of the cells with an inhibitor to block the gap junction-mediated intercellular communication. These results indicate that the bystander effect plays an important role in both the induction of a protective effect by the conditioning dose and the detrimental effect of the challenge irradiation. Gap junction-mediated intercellular communication was suggested to be involved in the induction of the bystander effect.


[Reasons for using emergency departments of major hospitals in Greater Tunis].
Tunisia, similar to many countries, has a problem of overcrowding of the emergency departments (ED). This study aimed to analyse the reasons for using EDs, and to describe the seriousness of the attendees' condition and their itinerary before their arrival at ED. This cross-sectional study in 2009 was conducted in ED of 4 hospitals in GreaterTunis and targeted 1058 patients of both sexes, aged 18 years, with stratification according to time of day of presentation to ED (morning, afternoon and evening). Information was recorded on the sociodemographic characteristics of the study participants, reasons for choosing ED, time of and reason for consultation, diagnosis and severity of illness. Over half the patients (52.5%) were male and the mean age was 46.0 (SD 18.1) years. The main reasons for choosing the ED were: speed (54.0%) and ease of access (47.7%) of ER and occurrence of an acute episode (26.4%). Patients with serious illness accounted for only 6.3% of those interviewed. Implementation of good practices and better coordination between public and private services and the ED are needed to reduce unnecessary visits to ED.


The role of radical prostatectomy and lymph node dissection in lymph node-positive prostate cancer: a systematic review of the literature.
Because pelvic lymph node (LN)-positive prostate cancer (PCa) is generally considered a regionally metastatic disease, surgery needs to be better defined. To review the impact of radical prostatectomy (RP) and pelvic lymph node dissection (PLND), possibly in conjunction with a multimodal approach using local radiotherapy and/or androgen-deprivation therapy (ADT), in LN-positive PCa. A systematic Medline search for studies reporting on treatment regimens and outcomes in patients with LN-positive PCa undergoing RP between 1993 and 2012 was performed. RP can improve progression-free and overall survival in LN-positive PCa, although there is a lack of high-level evidence. Therefore, the former practice of aborting surgery in the presence of positive nodes might no longer be supported by current evidence, especially in those patients with a limited LN tumor burden. Current data demonstrate that the lymphatic spread takes an ascending pathway from the pelvis to the retroperitoneum, in which the internal and the common iliac nodes represent critical landmarks in the metastatic distribution. Sophisticated imaging technologies are still under investigation to improve the prediction of LN-positive PCa. Nonetheless, extended PLND including the common iliac arteries should be offered to intermediate- and high-risk patients to improve nodal staging with a possible benefit in prostate-specific antigen progression-free survival by removing significant metastatic load. Adjuvant ADT has the potential to improve overall survival after RP; the therapeutic role of a trimodal approach with adjuvant local radiotherapy awaits further elucidation. Age is a critical parameter for survival because cancer-specific mortality exceeds overall mortality in younger patients (<60 yr) with high-risk PCa and should be an impetus to treat as thoroughly as possible. Increasing evidence suggests that RP and extended PLND improve survival in LN-positive PCa. Our understanding of surgery of the primary tumor in LN-positive PCa needs a conceptual change from a palliative option to the first step in a multimodal approach with a significant improvement of long-term survival and cure in selected patients.


3,4-Dichlorobenzyloxyacetic acid is extensively metabolized to a taurine conjugate in rats.
The metabolism of the antisickling agent 3,4-dichlorobenzyloxyacetic acid (3,4-DCBAA) was examined after ip administration to rats. Within 5 days after administration of radiolabeled 3,4-DCBAA, 77.4 +/- 4.6% of the dose was recovered in the urine and only 3.2 +/- 0.5% was recovered in the feces. Metabolites in the urine were isolated and characterized by HPLC, electron impact MS, and LC/MS, and their identities were confirmed by comparing their spectra with those of synthetic standards. Quantitation of these urinary metabolites revealed that the majority of the radioactive dose was excreted as a taurine conjugate (60.1 +/- 4.4%), while lesser amounts were excreted as 3,4-dichlorohippurate, unchanged 3,4-DCBAA, the glycine conjugate of 3,4-DCBAA, and a polar unknown which is believed to be glycolic acid. A pathway involving an initial O-dealkylation at the benzyl carbon of 3,4-DCBAA and proceeding through the glycine conjugation of 3,4-dichlorobenzoic acid has been proposed to explain the formation of 3,4-dichlorohippurate and the polar unknown. The extensive conjugation of 3,4-DCBAA with taurine is an unprecedented observation in rats, which usually utilize glycine for amino acid conjugation reactions. Further studies with 3,4-DCBAA may provide insight into the enzymatic mechanisms of taurine conjugation, which are not well defined at this time.


Molecular recognition and docking algorithms.
Molecular docking is an invaluable tool in modern drug discovery. This review focuses on methodological developments relevant to the field of molecular docking. The forces important in molecular recognition are reviewed and followed by a discussion of how different scoring functions account for these forces. More recent applications of computational chemistry tools involve library design and database screening. Last, we summarize several critical methodological issues that must be addressed in future developments.


Prognostic value and therapeutic potential of TREM-1 in Streptococcus pyogenes- induced sepsis.
TREM-1 (triggering receptor expressed on myeloid cells) is a surface molecule expressed on neutrophils and macrophages which has been implicated in the amplification of inflammatory responses triggered during infection. In the present study, we have investigated the clinical significance of TREM-1 in Streptococcus pyogenes-induced severe sepsis in both experimentally infected mice as well as in patients with streptococcal toxic shock. We found that S. pyogenes induced a dose-dependent upregulation of TREM-1 in in vitro cultured phagocytic cells and in the organs of S. pyogenes-infected mice. Furthermore, we reported a positive correlation between serum levels of soluble TREM-1 (sTREM-1) and disease severity in infected patients as well as in experimentally infected mice. Hence, sTREM-1 may represent a useful surrogate marker for streptococcal sepsis. We found that modulation of TREM-1 by administration of the TREM-1 decoy receptor rTREM-1/Fc substantially attenuated the synthesis of inflammatory cytokines. More importantly, treatment of S. pyogenes-infected septic mice with rTREM-1/Fc or the synthetically produced conserved extracellular domain LP17 significantly improved disease outcome. In summary, our data suggest that TREM-1 may not only represent a valuable marker for S. pyogenes infection severity but it may also be an attractive target for the treatment of streptococcal sepsis.


The role of PIM1 in the ibrutinib-resistant ABC subtype of diffuse large B-cell lymphoma.
Diffuse large B cell lymphoma (DLBCL) is a heterogeneous lymphoma and the most common subtype of non-Hodgkin lymphoma, accounting for roughly 30% of newly diagnosed cases in the United States. DLBCL can be separated into the activated B cell-like (ABC) and germinal center B cell-like (GCB) subtypes, with distinct gene expression profiles, oncogenic aberrations, and clinical outcomes. ABC-DLBCL is characterized by chronically active B-cell receptor (BCR) signaling that can be modulated by Bruton's tyrosine kinase (BTK) activity. Thus, BTK serves as an attractive therapeutic target in this type of B-cell malignancy. Ibrutinib, a first-in-class, orally available covalent BTK inhibitor, has demonstrated clinical activity in several B-cell leukemias and lymphomas. A phase 1/2 clinical trial of single-agent ibrutinib in relapsed and refractory DLBCL patients revealed an overall response rate of 37% in ABC-DLBCL patients. However, responses to kinase-directed therapies are often limited by emerging resistance mechanisms that bypass the therapeutic target. Here we report the discovery of point mutations within the kinase PIM1 that reduce sensitivity to ibrutinib in ABC-DLBCL. These mutations stabilize PIM1 and affect upstream regulators and downstream targets of NF-κB signaling. The introduction of mutant PIM1 into an ABC-DLBCL cell line, TMD8, increased colony formation and decreased sensitivity to ibrutinib. In addition, ibrutinib-resistant cell lines generated by prolonged ibrutinib exposure in vitro upregulated PIM1 expression, consistent with a role for PIM1 in antagonizing ibrutinib activity. The combination of a pan-PIM inhibitor with ibrutinib synergistically inhibited proliferation in vitro and tumor growth in vivo. Together, these data provide a rationale for combining BTK and PIM1 inhibition in the treatment of ABC-DLBCL.


Performance and safety of holmium: YAG laser optical fibers.
Lower-pole ureteronephroscopy requires transmission of holmium:YAG energy along a deflected fiber. Current ureteroscopes are capable of high degrees of deflection, which may stress laser fibers beyond safe limits during lower-pole use. We hypothesized that optical fiber and safety measures differ among manufacturers. Small (200-273-microm) and medium-diameter (300-400-microm) Ho:YAG fibers were tested in a straight and 180 degrees bent configuration. Energy transmission was measured by an energy detector. Fiber durability was assessed by firing the laser in sequentially tighter bending diameters. The fibers were bent to 180 degrees with a diameter of 6 cm and run at 200- to 4000-mJ pulse energy to determine the minimum energy required to fracture the fiber. The bending diameter was decreased by 1-cm increments and testing repeated until a bending diameter of 1 cm was reached. The maximum deflection of the ACMI DUR-8E ureteroscope with each fiber in the working channel was recorded. The flow rate through the working channel of the DUR-8E was measured for each fiber. The mean energy transmission differed among fibers (P < 0.001). The Lumenis SL 200 and the InnovaQuartz 400 were the best small and medium-diameter fibers, respectively, in resisting thermal breakdown (P < 0.01). The Dornier Lightguide Super 200 fractured repeatedly at a bend diameter of 2 cm and with the lowest energy (200 mJ). The other small fibers fractured only at a bend diameter of 1 cm. The Sharplan 200 and InnovaQuartz Sureflex 273T were the most flexible fibers, the Lumenis SL 365 the least. The flow rate was inversely proportional to four times the power of the diameter of the fiber. Optical performance and safety differ among fibers. Fibers transmit various amounts of energy to their cladding when bent. During lower-pole nephroscopy with the fiber deflected, there is a risk of fiber fracture from thermal breakdown and laser-energy transmission to the endoscope. Some available laser fibers carry a risk of ureteroscope damage.


Uroperitoneum in the foal.
The case records of 22 foals with uroperitoneum were reviewed. The most common cause was a defect in the urinary bladder. A sex predilection for males was apparent. Clinical signs were tachycardia, tachypnea, and abdominal distention. The history pertaining to micturition was variable. Hyponatremia, hypochloremia, and hyperkalemia were marked. A reliable means of diagnosis was considered to be the demonstration of a wide disparity in serum and peritoneal fluid creatinine values. Surgical repair was successful in 11 of 18 cases, although surgical and anesthetic complications were common. Some of the defects were believed to be congenital.


Lipid metabolism-related gene prognostic index (LMRGPI) reveals distinct prognosis and treatment patterns for patients with early-stage pulmonary adenocarcinoma.
Background: Lipid metabolism plays a pivotal role in cancer progression and metastasis. This study aimed to investigate the prognostic value of lipid metabolism-related genes (LMRGs) in early-stage lung adenocarcinoma (LUAD) and develop a lipid metabolism-related gene prognostic index (LMRGPI) to predict their overall survival (OS) and treatment response. Methods: A total of 774 early-stage LUAD patients were identified from The Cancer Genome Atlas (TCGA, 403 patients) database and Gene Expression Omnibus (GEO, 371 patients) database. The non-negative Matrix Factorization (NMF) algorithm was used to identify different population subtypes based on LMRGs. The Least Absolute Shrinkage and Selection Operator (LASSO) and multivariate Cox regression analyses were used to develop the LMRGPI, with receiver operating characteristic (ROC) curves and concordance index being used to evaluate its performance. The characteristics of mutation landscape, enriched pathways, tumor microenvironment (TME), and treatment response between different LMRGPI groups were also investigated. Results: We identified two population subtypes based on LMRGs in the TCGA-LUAD cohort, with distinct prognosis, TME, and immune status being observed. LMRGPI was developed based on the expression levels of six LMRGs, including ANGPTL4, NPAS2, SLCO1B3, ACOXL, ALOX15, and B3GALNT1. Higher LMRGPI was correlated with poor OS both in TCGA and GSE68465 cohorts. Two nomograms were established to predict the survival probability of early-stage LUAD, with higher consistencies being observed between the predicted and actual OS. Higher LMRGPI was significantly correlated with more frequent TP53 mutation, higher tumor mutation burden (TMB), and up-regulation of CD274. Besides, patients with higher LMRGPI presented unremarkable responses for gefitinib, erlotinib, cisplatin, and vinorelbine, while they tend to have a favorable response for immune checkpoint inhibitors (ICIs). The opposite results were observed in the low-LMRGPI group. Conclusions: We comprehensively investigated the prognostic value of LMRGs in early-stage LUAD. Given its good prognostic ability, LMRGPI could serve as a promising biomarker to predict the OS and treatment response of these patients.


Prognostic Genetic Markers for Thrombosis in COVID-19 Patients: A Focused Analysis on D-Dimer, Homocysteine and Thromboembolism.
COVID-19 is caused by Severe Acute Respiratory Syndrome Coronavirus-2, which has infected over thirty eight million individuals worldwide. Emerging evidence indicates that COVID-19 patients are at a high risk of developing coagulopathy and thrombosis, conditions that elevate levels of D-dimer. It is believed that homocysteine, an amino acid that plays a crucial role in coagulation, may also contribute to these conditions. At present, multiple genes are implicated in the development of these disorders. For example, single-nucleotide polymorphisms (SNPs) in FGG, FGA, and F5 mediate increases in D-dimer and SNPs in ABO, CBS, CPS1 and MTHFR mediate differences in homocysteine levels, and SNPs in TDAG8 associate with Heparin-induced Thrombocytopenia. In this study, we aimed to uncover the genetic basis of the above conditions by examining genome-wide associations and tissue-specific gene expression to build a molecular network. Based on gene ontology, we annotated various SNPs with five ancestral terms: pulmonary embolism, venous thromboembolism, vascular diseases, cerebrovascular disorders, and stroke. The gene-gene interaction network revealed three clusters that each contained hallmark genes for D-dimer/fibrinogen levels, homocysteine levels, and arterial/venous thromboembolism with F2 and F5 acting as connecting nodes. We propose that genotyping COVID-19 patients for SNPs examined in this study will help identify those at greatest risk of complications linked to thrombosis.


Synthesis and biology of the conformationally restricted ACPD analogue, 2-aminobicyclo[2.1.1]hexane-2,5-dicarboxylic acid-I, a potent mGluR agonist.
To better characterize the roles of metabotropic glutamate receptors (mGluRs) in physiological and pathophysiological processes, there is an important need to learn more about the structural features relevant to the design of novel, high-affinity ligands that are family and subtype specific. To date, many of the biological studies that have been conducted in the area of mGluR research have made use of the agonist (1S,3R)-ACPD. This compound has been shown to act as an agonist at both the group I and group II receptors while showing little selectivity among the four subtypes belonging to these two groups. Moreover, (1S,3S)-ACPD, the cis isomer, shows negligible activity at group I receptors and is a good agonist of mGluR2. Since ACPD is itself somewhat flexible, with four distinctive conformations being identified from molecular modeling studies for the trans isomer and five conformations for the cis isomer, we believed that it would be of interest to examine the activity of an ACPD analogue that has been constrained through the introduction of a single carbon atom bridge. Accordingly, we have prepared an aminobicyclo[2.1.1]hexanedicarboxylic acid (ABHxD-I) analogue of ACPD. The synthesis of this compound was accomplished by use of an intramolecular [2 + 2] photocycloaddition reaction, in which four distinct isomers were isolated. Of these four compounds, only a single isomer, ABHxD-I (6a), was found to be a potent agonist of the mGluRs. This compound, which expresses the fully extended glutamate conformation, was found to be more potent than ACPD at all six of the eight mGluR subtypes that were investigated and to be comparable to or more potent than the endogenous ligand, glutamate, for these receptors. Interestingly, despite its fixed conformation, ABHxD-I, like glutamate, shows little subtype selectivity. Through modeling studies of ABHxD-I (6a), ABHD-VI, LY354740, (1S,3R)-ACPD, (1S, 3S)-ACPD, and l-glutamate, we conclude that the aa conformation of l-glutamate is the active conformation for both group I and group II mGluRs. Moreover, the modeling-based comparisons of these ligands suggest that the selectivity exhibited by LY354740 between the group I and group II mGluRs is not a consequence of different conformations of L-glutamate being required for recognition at these mGluRs but rather is related to certain structural elements within certain regions having a very different impact on the group I and group II mGluR activity. The enhanced potency of ABHxD-I relative to trans-ACPD commends it as a useful starting point in the design of subtype selective mGluR ligands.


Restrictions on rotational and translational diffusion of pigment in the membranes of a rhabdomeric photoreceptor.
Individual, isolated rhabdoms from dark-adapted crayfish (Orconectes, Procambarus) were studied with a laterally incident microbeam that could be placed in single stacks of microvilli. Concentration gradients of metarhodopsin along the lengths of microvilli were produced by local bleaches, accomplished by irradiation with small spots of orange light at pH 9 in the presence of glutaraldehyde or formaldehyde. No subsequent redistribution of pigment was observed in the dark, indicating an absence of translational diffusion. On the basis of comparison with other systems, glutaraldehyde, but not formaldehyde (0.75%), would be expected to prevent diffusion of protein in the membrane. Under the same conditions photodichroism is observed, indicating an absence of free Brownian rotation. Photodichroism is larger in glutaraldehyde than in formaldehyde, suggesting that the bifunctional reagent quiets some molecular motion that is present after treatment with formaldehyde. Quantitative comparison of photodichroism with mathematical models indicates that the pigment absorption vectors are aligned within +/- 50 degrees of the microvillar axes and are tilted into the surface of the membrane at an average value of about 20 degrees. The photoconversion of rhodopsin to metarhodopsin is accompanied by an increase in molar extinction of about 20% at the lambda maxand a reorientation of the absorption vector by several degrees. The transition moment either tilts further into the membrane or loses some of its axial orientation, or both. The change in orientation is 3.5 time larger in formaldehyde than in glutaraldehyde.


Monitoring microbiological changes in drinking water systems using a fast and reproducible flow cytometric method.
Flow cytometry (FCM) is a rapid, cultivation-independent tool to assess and evaluate bacteriological quality and biological stability of water. Here we demonstrate that a stringent, reproducible staining protocol combined with fixed FCM operational and gating settings is essential for reliable quantification of bacteria and detection of changes in aquatic bacterial communities. Triplicate measurements of diverse water samples with this protocol typically showed relative standard deviation values and 95% confidence interval values below 2.5% on all the main FCM parameters. We propose a straightforward and instrument-independent method for the characterization of water samples based on the combination of bacterial cell concentration and fluorescence distribution. Analysis of the fluorescence distribution (or so-called fluorescence fingerprint) was accomplished firstly through a direct comparison of the raw FCM data and subsequently simplified by quantifying the percentage of large and brightly fluorescent high nucleic acid (HNA) content bacteria in each sample. Our approach enables fast differentiation of dissimilar bacterial communities (less than 15 min from sampling to final result), and allows accurate detection of even small changes in aquatic environments (detection above 3% change). Demonstrative studies on (a) indigenous bacterial growth in water, (b) contamination of drinking water with wastewater, (c) household drinking water stagnation and (d) mixing of two drinking water types, univocally showed that this FCM approach enables detection and quantification of relevant bacterial water quality changes with high sensitivity. This approach has the potential to be used as a new tool for application in the drinking water field, e.g. for rapid screening of the microbial water quality and stability during water treatment and distribution in networks and premise plumbing.


Highly pathogenic avian influenza subtype H5Nx clade 126.96.36.199 outbreaks in Dutch poultry farms, 2014-2018: Clinical signs and mortality.
In recent years, different subtypes of highly pathogenic avian influenza (HPAI) viruses caused outbreaks in several poultry types worldwide. Early detection of HPAI virus infection is crucial to reduce virus spread. Previously, the use of a mortality ratio threshold to expedite notification of suspicion in layer farms was proposed. The purpose of this study was to describe the clinical signs reported in the early stages of HPAI H5N8 and H5N6 outbreaks on chicken and Pekin duck farms between 2014 and 2018 in the Netherlands and compare them with the onset of an increased mortality ratio (MR). Data on daily mortality and clinical signs from nine egg-producing chicken farms and seven Pekin duck farms infected with HPAI H5N8 (2014 and 2016) and H5N6 (2017-2018) in the Netherlands were analysed. In 12 out of 15 outbreaks for which a MR was available, MR increase preceded or coincided with the first observation of clinical signs by the farmer. In one chicken and two Pekin duck outbreaks, clinical signs were observed prior to MR increase. On all farms, veterinarians observed clinical signs of general disease. Nervous or locomotor signs were reported in all Pekin duck outbreaks, but only in two chicken outbreaks. Other clinical signs were observed less frequently in both chickens and Pekin ducks. Compared to veterinarians, farmers observed and reported clinical signs, especially respiratory and gastrointestinal signs, less frequently. This case series suggests that a MR with a set threshold could be an objective parameter to detect HPAI infection on chicken and Pekin duck farms at an early stage. Observation of clinical signs may provide additional indication for farmers and veterinarians for notifying a clinical suspicion of HPAI infection. Further assessment and validation of a MR threshold in Pekin ducks are important as it could serve as an important tool in HPAI surveillance programs.


Intraruminal administration of milk in the calf as a model for ruminal drinking: morphological and enzymatical changes in the jejunal mucosa.
In order to develop a calf model for studying the syndrome of ruminal drinking (RD) in veal calves, three dual-fistulated calves were used to test the effect of intraruminal administration of milk replacer on the jejunal mucosa. Biopsies of the proximal jejunal mucosa were taken through a jejunal fistula and the mucosal morphology and the activities of two brush border enzymes, lactase and alkaline phosphatase, were determined. Means of villus length and brush border enzyme activities decreased during the period of intraruminal administration of milk. The hyperplastic villus atrophy in this model was similar to that found in chronic RD patients in previous studies. This could not be associated with isolation of pathogenic micro-organisms from the faeces and is probably the consequence of the intraruminal milk feeding procedure itself. Clinical recovery from the signs of RD occurred rapidly after intraruminal administration of milk ceased and was followed by restoration of villus length and brush border enzyme activities 3-4 weeks later.


Quantitative vertebral morphometry using neighbor-conditional shape models.
A novel method for vertebral fracture quantification from X-ray images is presented. Using pairwise conditional shape models trained on a set of healthy spines, the most likely normal vertebra shapes are estimated conditional on the shapes of all other vertebrae in the image. The difference between the true shape and the reconstructed normal shape is subsequently used as a measure of abnormality. In contrast with the current (semi-)quantitative grading strategies this method takes the full shape into account, it develops a patient-specific reference by combining population-based information on biological variation in vertebral shape and vertebra interrelations, and it provides a continuous measure of deformity. The method is demonstrated on 282 lateral spine radiographs with in total 93 fractures. Vertebral fracture detection is shown to be in good agreement with semi-quantitative scoring by experienced radiologists and is superior to the performance of shape models alone.


Advances in hip arthroplasty surgery: what is justified?
Total hip arthroplasty (THA) surgery has shown dramatic changes in terms of increased number of procedures and of technical development in recent years. It has been described as "the operation of the 20th century" for the excellent results, the high satisfaction of the patients and the improvement of the quality of life.A lot of variations have been introduced over the last few decades in THA especially in terms of indications (both in younger and older patients), techniques and devices (approaches, tissue preservation, biomaterials and industrial finishing), per-operative management (blood loss and pain control) and post-operative protocols (the so called "fast track" surgery). Looking at all these advances the emerging question is: have all of them been justified both in terms of improvement of the results for the patients and of the cost/benefit ratio from an economical point of view?The purpose of this paper is to critically analyse the advantages and the disadvantages of the theoretically proposed "advances in hip arthroplasty" and attempt to understand which are justified of such "advances" nowadays, based on the international and the European perspective with a focus on the author's personal clinical experience. Cite this article: EFORT Open Rev 2017;2. DOI: 10.1302/2058-5241.2.170008. Originally published online at www.efortopenreviews.org.


Steady-state kinetic analysis of the quinoprotein methylamine dehydrogenase from Paracoccus denitrificans.
A steady-state kinetic analysis was performed of the reaction of methylamine and phenazine ethosulphate (PES) with the quinoprotein methylamine dehydrogenase from Paracoccus denitrificans. Experiments with methylamine and PES as varied-concentration substrates produced a series of parallel reciprocal plots, and when the concentrations of these substrates were varied in a constant ratio a linear reciprocal plot of initial velocity against PES concentration was obtained. Nearly identical values of V/Km of PES were obtained with four different n-alkylamines. These data suggest that this reaction proceeds by a ping-pong type of mechanism. The enzyme reacted with a variety of n-alkylamines but not with secondary, tertiary or aromatic amines or amino acids. The substrate specificity was dictated primarily by the Km value exhibited by the particular amine. A deuterium kinetic isotope effect was observed with deuterated methylamine as a substrate. The enzyme exhibited a pH optimum for V at pH 7.5. The absorbance spectrum of the pyrroloquinoline quinone prosthetic group of this enzyme was also effected by pH at values greater than 7.5. The enzyme was relatively insensitive to changes in ionic strength, and exhibited a linear Arrhenius plot over a range of temperatures from 10 degrees C to 50 degrees C with an energy of activation 46 kJ/mol (11 kcal/mol).


Biologic agents for severe asthma patients: clinical perspectives and implications.
Asthma is a chronic inflammatory multifactorial disorder of the airways characterized by the involvement of immune cells and mediators in its onset and maintenance. Traditional therapeutic strategies have been unsatisfactory in controlling the underlying pathology, especially in the more severe states. Hence in the last couple of decades, new biological approaches targeting molecular mediators have been developed. In this narrative review we examine biological agents currently available for the management of severe asthma, focusing our attention on their clinical application, pros and cons, and in particular on gaps regarding the use of these agents. The most well-known and used biologic agent in clinical practice is omalizumab, though there is emerging evidence for mepolizumab too. The future of these biological therapies is to broaden our knowledge of their practical use and ascertain predictive biomarkers, or define an algorithm, useful in the optimal application of these 'biological weapons'.


Lymph node topology dictates T cell migration behavior.
Adaptive immunity is initiated by T cell recognition of foreign peptides presented on dendritic cells (DCs) by major histocompatibility molecules. These interactions take place in secondary lymphoid tissues, such as lymph nodes (LNs) and spleen, and hence the anatomical structure of these tissues plays a crucial role in the development of immune responses. Two-photon microscopy (2PM) imaging in LNs suggests that T cells walk in a consistent direction for several minutes, pause briefly with a regular period, and then take off in a new, random direction. Here, we construct a spatially explicit model of T cell and DC migration in LNs and show that all dynamical properties of T cells could be a consequence of the densely packed LN environment. By means of 2PM experiments, we confirm that the large velocity fluctuations of T cells are indeed environmentally determined rather than resulting from an intrinsic motility program. Our simulations further predict that T cells self-organize into microscopically small, highly dynamic streams. We present experimental evidence for the presence of such turbulent streams in LNs. Finally, the model allows us to estimate the scanning rates of DCs (2,000 different T cells per hour) and T cells (100 different DCs per hour).


Positive youth development programmes to reduce substance use in young people: Systematic review.
Substance use has detrimental short-term and long-term consequences for young people. Positive youth development (PYD) interventions, which favour promotion of positive assets over traditional risk reduction, have received attention recently as a possible intervention to prevent adolescent substance use. We aimed to synthesise the evidence on PYD interventions for reduction in substance use in young people. We searched 21 databases, including MEDLINE, PsycINFO, CINAHL and CENTRAL, and hand-searched key journals and websites. We included studies with more than half of participants aged 11-18 years where interventions meeting a pre-specified definition of PYD were delivered in community settings outside of normal school hours and did not target parents or young people with pre-defined conditions. Two reviewers screened records, assessed full-text studies for inclusion, and extracted data. A modified Cochrane risk of bias tool was used for quality assessment. Ten studies reported in 13 reports were included in our synthesis. PYD interventions did not have an effect of statistical or public health significance on any substance use, illicit drug use or alcohol outcomes in young people. Interventions were diverse in content and delivery. Our review suggests that existing PYD interventions subject to evaluation do not appear to have produced reductions in substance use of public health significance. However, these interventions may not be the best exemplars of a PYD approach. Therefore, our findings should not be taken as evidence for the ineffectiveness of PYD as a theory of change for reducing substance use among young people. Additional rigorous evaluation of PYD interventions is key before further investment. Evaluations were of highly variable quality. Though searches were extensive, we were unable to test for publication bias.


Antimicrobial drug resistance of Salmonella and Escherichia coli isolates from cattle feces, hides, and carcasses.
To determine patterns of cross-contamination and antibiotic susceptibility of microorganisms commonly associated with cattle, 60 cattle shipped to a commercial abattoir (20 in each of three separate trial periods) were followed through processing. Samples for bacterial isolation were collected from the feces and hides immediately before shipping, from the hides at the abattoir after exsanguination, and from the carcasses before evisceration and in the cooler. Samples were cultured for Salmonella and non-type-specific Escherichia coli. Salmonella was identified in 33.9% (n = 20) of the fecal samples and on 37.3% (n = 22) of the hides before shipment. At the abattoir, the proportion of hides from which Salmonella was isolated increased (P < 0.001) to 84.2% (48 hides). Nonspecific E. coli and Salmonella were recovered from 40.4 and 8.3% of preevisceration carcass samples, respectively. No Salmonella or nonspecific E. coli were recovered from hotbox carcass samples. Isolates were tested for antimicrobial drug susceptibility. For nonspecific E. coli, 80.3% (n = 270) of the isolates were resistant to at least one antimicrobial drug. For Salmonella, 97% (n = 101) of the isolates were resistant to at least one antimicrobial drug; however, only 4.0% were resistant to two or more. The most common resistance was to sulfamethoxazole. These results indicate that the presence of microorganisms resistant to antimicrobial drugs is common in cattle and beef. Further studies are needed to identify the sources and causes of this drug resistance.


Does the spinal cord generate functionally significant sympathetic activity in the awake rat?
Previous studies reach conflicting conclusions regarding the presence of physiologically significant sympathetic nerve activity (SNA) in conscious rats with transection of the cervical spinal cord (CST). The objective of the current study was to determine whether either spinally generated SNA or nonspecific effects of antagonists are responsible for the natriuresis and decreased heart rate that accompany adrenergic blockade in CST rats. To test the first possibility, adrenergic antagonists (phentolamine and propranolol) were administered to three groups of CST rats: 1) renal denervated, 2) adrenalectomized or sham-adrenalectomized, and 3) rats who received the ganglionic blocker hexamethonium. Neither renal denervation, adrenalectomy, nor ganglionic blockade prevented either the three- to fivefold increase in sodium excretion or the 25-50 beats/min decrease in heart rate previously reported. We then administered combinations of different adrenergic antagonists to CST rats to test for nonspecific effects of the drugs. Whereas idazoxan+propranolol reproduced the natriuresis seen with phentolamine+propranolol, prazosin+yohimbine+propranolol did not. We conclude that there is no evidence for functionally significant spinally generated SNA in conscious CST rats and that the natriuresis observed with phentolamine administration is due to imidazoline binding.


Localized intradermal microinjection of tranexamic acid for treatment of melasma in Asian patients: a preliminary clinical trial.
Melasma is a common cosmetic problem among Asians. While various treatments are currently being used, there is no entirely satisfactory treatment. It was recently reported that the topical plasmin inhibitor is an effective treatment for ultraviolet-induced hyperpigmentation. Because there are no studies assessing the efficacy and safety of localized microinjection of tranexamic acid (TA) for the treatment of melasma, we conducted a pilot study to evaluate the efficacy and side effects of this potentially new method for the treatment of melasma in Korean women. A total of 100 women with melasma, after written consent, were enrolled for a prospective open pilot study of 12 weeks. After applying topical anesthesia, 0.05 mL TA (4 mg/mL) was injected intradermally into the melasma lesion at 1 cm intervals by using a 0.5 mL insulin syringe with a 30-gauge needle. This was repeated weekly for 12 weeks. A clinical investigator evaluated the results by using the Melasma Area and Severity Index (MASI) at baseline and at 4, 8, and 12 weeks. The patient satisfaction questionnaire was documented at 12 weeks. Safety evaluations were performed at each follow-up visit. Eighty-five patients completed the trial. A significant decrease in the MASI from baseline to 8 and 12 weeks was observed (13.22+/-3.02 vs 9.02+/-2.62 at week 8 and vs. 7.57+/-2.54 at week 12; p<.05 for both). The patients' self-assessment of melasma improvement was as follows: 8 of 85 patients (9.4%) rated as good (51-75% lightening), 65 patients (76.5%) as fair (26-50% lightening), and 12 patients (14.1%) as poor (0-25% lightening). Side effects were minimal and all the patients tolerated the treatment well. Based on these results, we suggest that the intralesional localized microinjection of TA acid can be used as a potentially new, effective, and safe therapeutic modality for the treatment of melasma.


Histidine-Tryptophan-Ketoglutarate (HTK) is associated with reduced graft survival in deceased donor livers, especially those donated after cardiac death.
Single-center studies have reported that liver allograft survival is not affected by preservation in histidine-tryptophan-ketoglutarate (HTK) versus University of Wisconsin (UW) solution. We analyzed the UNOS database of liver transplants performed from July, 2004, through February, 2008, to determine if preservation with HTK (n = 4755) versus UW (n = 12 673) impacted graft survival. HTK preservation of allografts increased from 16.8% in 2004 to 26.9% in 2008; this was particularly striking among donor after cardiac death (DCD) allografts, rising from 20.7% in 2004 to 40.9% in 2008. After adjusting for donor, recipient and graft factors that affect graft survival, HTK preservation was associated with an increased risk of graft loss (HR 1.14, p = 0.002), especially with DCD allografts (HR 1.44, P = 0.025) and those with cold ischemia time over 8 h (HR 1.16, P = 0.009). Furthermore, HTK preservation was associated with a 1.2-fold higher odds of early (< 30 days) graft loss as compared to UW preservation (OR 1.20, p = 0.012), with a more pronounced effect on allografts with cold ischemia time over 8 h (OR 1.31, p = 0.007), DCD allografts (OR 1.63, p = 0.09) and donors over 70 years (OR 1.67, p = 0.081). These results suggest that the increasing use of HTK for abdominal organ preservation should be reexamined.


Trace element status in type 2 diabetes: A meta-analysis.
Type 2 diabetes is a chronic metabolic disorder that has been associated with alterations in the status of trace elements, including zinc, copper, iron and manganese. However, clinical studies reporting statuses of these trace elements in type 2 diabetes patients compared to controls have shown conflicting results. This meta-analysis aimed to summarize the existing literature on the statuses of zinc, copper, iron, and manganese in Type 2 diabetes mellitus patients. A literature search of Embase, PubMed, EBSCOHost, ScienceDirect, Scopus, Cochrane library and Web of Science electronic databases was conducted to find studies published from 1970 to November 2016 that compared the trace elements of interest between type 2 diabetic patients and healthy controls. Keywords used were type 2 diabetes, diabetes, hyperglycemia, insulin, glucose, HbA1c, trace elements, micronutrients, zinc, manganese, copper, ceruloplasmin, iron and ferritin. The bias corrected Hedges' g, was utilized as the effect sizes. Due to the biological interaction between trace elements, it is important to collectively evaluate the statuses of these minerals in type 2 diabetes. Thus, the robust variance estimation method was chosen to handle dependency between multiple outcomes. A total of 52 studies met the inclusion criteria, amounting to 98 effect sizes. Diabetic patients (n=20183) had significantly lower zinc status when compared to controls (effect size = -1.73, p<0.01); whereas copper (effect size = 1.10, p<0.05) and ferritin levels (effect size = 1.05, p<0.01) were significantly higher. Although not significant, ceruloplasmin (effect size = 1.85, p=0.06) and iron (effect size = 1.42, p=0.06) levels were higher, and manganese (effect size = 0.27, p=0.34) was lower in patients. Results from this meta-analysis indicate lower zinc status accompanied by increased copper and ferritin levels in patients with type 2 diabetes when compared to controls.


Cigarette smoking and esophageal cancer risk: an evaluation based on a systematic review of epidemiologic evidence among the Japanese population.
Although cigarette smoking is considered as an important risk factor for esophageal cancer, the magnitude of the association might be varied among geographic areas. Therefore, we reviewed epidemiologic studies on the association between cigarette smoking and esophageal cancer among the Japanese population. Original articles were obtained from MEDLINE searched using PubMed or from searches of the Ichushi database, complemented by manual searches. Evaluation of associations was based on the strength of evidence ('convincing', 'probable', 'possible' or 'insufficient') and the magnitude of association ('strong', 'moderate', 'weak' or 'no association'), together with biological plausibility as previously evaluated by the International Agency for Research on Cancer. We identified four cohort studies and 11 case-control studies. All cohort studies and eight case-control studies showed strong positive associations between esophageal cancer and cigarette smoking. All cohort studies and five case-control studies showed that cigarette smoking had dose-response relationships with esophageal cancer. Meta-analysis of 12 studies indicated that the summary estimate for ever smokers relative to never smokers was 3.01 (95% confidence interval: 2.30-3.94). Summary relative risk for current and former smokers relative to never smokers was 3.73 (2.16-6.43) and 2.21 (1.60-3.06), respectively. We conclude that there is convincing evidence that cigarette smoking strongly increases the risk of esophageal cancer in the Japanese population.


Apremilast interferes with the TGFβ1-induced transition of human skin fibroblasts into profibrotic myofibroblasts: in vitro study.
Fibroblast-to-myofibroblast transition and extracellular matrix overproduction represent progressive events in chronic inflammatory and fibrotic diseases, in which TGFβ1 is one of the key mediators. Phosphodiesterase 4 (PDE4) acts as a proinflammatory enzyme through the degradation of cyclic adenosine monophosphate and it is overexpressed in skin fibroblasts. The study investigated how apremilast (a PDE4 inhibitor) interferes with the intracellular signalling pathways responsible for the TGFβ1-induced fibroblast-to-myofibroblast transition and profibrotic extracellular matrix protein synthesis. Cultured human skin fibroblasts were stimulated with TGFβ1 (10 ng/ml) alone or combined with apremilast (1 and 10 μM) for 4, 16 and 24 h. Other aliquots of the same cells were previously stimulated with TGFβ1 and then treated with apremilast (1 and 10 μM) for 4, 16 and 24 h, always under stimulation with TGFβ1. Gene and protein expression of αSMA, type I collagen (COL1) and fibronectin were evaluated, together with the activation of small mothers against decapentaplegic 2 and 3 (Smad2/3) and extracellular signal-regulated kinase (Erk1/2) proteins. Apremilast reduced the TGFβ1-induced increase in αSMA, COL1 and fibronectin gene expression at 4 and 16 h, and protein synthesis at 24 h of treatment in cultured fibroblasts, even for cells already differentiated into myofibroblasts by way of a previous stimulation with TGFβ1. Apremilast inhibited the TGFβ1-induced Smad2/3 and Erk1/2 phosphorylation at 15 and 30 min. Apremilast seems to inhibit in vitro the fibroblast-to-myofibroblast transition and the profibrotic activity induced by TGFβ1 in cultured human skin fibroblasts by downregulating Smad2/3 and Erk1/2 intracellular signalling pathways.


[Social pediatric screening of developmental status for school entry (SOPESS): validity in the domain of speech and language].
In addition to general cognitive and motor skills, the social-pediatric screening of developmental status for school entry (SOPESS) provides subtests for assessing speech and language in a differentiated way. In a special validation study, these domains are correlated to coextensive scales of SETK 3-5. The SOPESS features high specificity and results in reliable true negative findings. In addition, a preliminary evaluation of language skills considering migrant background is given. Children with an unsatisfactory status of language competence are treated separately in the SOPESS.


OCT-1 is over-expressed in intestinal metaplasia and intestinal gastric carcinomas and binds to, but does not transactivate, CDX2 in gastric cells.
Intestinal metaplasia (IM) is a preneoplastic lesion of the stomach in which there is transdifferentiation of the gastric mucosa to an intestinal phenotype. The caudal-related homeobox gene CDX2 encodes an intestine-specific transcription factor crucial for the regulation of proliferation and differentiation of intestinal cells. In addition, CDX2 is involved in the induction of IM in the stomach. The aim of this study was to access the putative involvement of OCT-1 in the induction of CDX2 expression de novo in gastric mucosa leading to the onset of IM. OCT-1 protein expression was evaluated by immunohistochemistry in 31 biopsies with chronic gastritis, 15 biopsies with foci of IM and adjacent gastric mucosa and 42 gastric carcinomas. Furthermore, we evaluated OCT-1 binding by electrophoretic mobility shift assay and activation of the CDX2 promoter by co-transfecting a CDX2 promoter/reporter construct with an OCT-1 expression vector in two gastric carcinoma cell lines, GP220 and MKN45. Our results show that OCT-1 is expressed in chronic gastritis, particularly when it is adjacent to IM and is expressed in 87% of IM foci. Furthermore, 74% of the gastric carcinomas were positive for OCT-1 and a strong association was observed between OCT-1 expression and intestinal-type carcinoma. We identified that OCT-1 binds to the CDX2 promoter, although we could not see a transactivation effect in gastric carcinoma cell lines. In conclusion, we observed increased OCT-1 expression in IM and in intestinal gastric carcinomas and identified the capacity of OCT-1 to bind to the CDX2 promoter, although we could not demonstrate a direct effect of OCT-1 in the transactivation of CDX2.


Staging and recurrence of disease in squamous cell carcinoma of the vulva.
In order to determine the prognostic significance of applying the revised FIGO staging system and identify factors contributing to survival after documentation of recurrent disease, a retrospective chart review of our vulvar cancer population was performed. Over a 17-year interval 135 patients were uniformly treated with primary surgical treatment consisting of radical vulvectomy and bilateral groin dissection. Factors contributing to disease-free survival were analyzed using a Cox proportional hazards model. Covariates of survival after recurrence of disease were analyzed using the log-rank method. Neither the clinical assessment of the groin nodes, nor the presence or absence of perineal involvement were related to outcome. Only lesion size and surgical status of the inguinal nodes were significant predictors of disease-free survival (P = 0.02 and P = 0.03, respectively). In addition, there was a statistically significant relationship between the extent of groin involvement (negative, unilateral positive, and bilateral positive nodes) and associated decrement in disease-free survival (P = 0.01). Thirty patients developed recurrence of disease from 2.0 to 47.3 months following surgery. The location of the recurrence, interval from primary therapy to recurrence, and status of the groin nodes at initial surgery were significant prognostic factors in subsequent survival. The revised staging system demonstrated an improvement in patient stratification compared to the criteria of the prior classification. The data are also consistent with the distinction made between Stage III and IV disease in the new classification. The status of the groin nodes at original surgery remained an important prognostic factor even in those patients who later demonstrated recurrence of disease.


Two sequences flanking the major autophosphorylation site of the insulin receptor are essential for tyrosine kinase activation.
The tyrosine kinase domain of the human insulin receptor (IR) contains several short amino acid motifs which are strictly conserved in all protein kinases and two sequence motifs which are specific to the tyrosine kinases (AAR or RAA and P(I)/VK/RWT/M). In the serine/threonine kinases these motifs are replaced by the sequences KPE and GT/SXXY/PX respectively. In the present work, the tyrosine kinase-specific sequences of the IR (1134AAR1136 and 1172PVRWM1176) were replaced using site-directed mutagenesis by sequences which confer a serine kinase specificity on the receptor. Five different IR mutants were expressed in Chinese hamster ovary (CHO) or COS cells and their structural and functional properties compared with those of the wild-type recombinant human IR. These mutants are processed normally and bind insulin with normal affinities. None of the mutants containing a putative serine kinase-specific sequence display detectable autophosphorylation or tyrosine kinase activity in response to insulin, either in vitro or in vivo. These mutants were also unable to phosphorylate serine/threonine kinase substrates after insulin stimulation. Unexpectedly, they showed impaired ATP binding, as studied by an original technique consisting of cross-linking adenosine 5'-([35S]thio)triphosphate to partially purified receptors. Finally, none of the studied mutants transmit the insulin signal necessary to stimulate either DNA or glycogen synthesis. These data provide evidence for the importance of these conserved sequences in the kinase domain for both receptor activation and kinase activity. Furthermore, they demonstrate that the exchange of sequences specific to the catalytic domain of tyrosine kinases for those specific to the serine/threonine kinases is not sufficient to confer serine/threonine specificity on the insulin receptor.


17p12 Influences Hematoma Volume and Outcome in Spontaneous Intracerebral Hemorrhage.
Hematoma volume is an important determinant of clinical outcome in spontaneous intracerebral hemorrhage (ICH). We performed a genome-wide association study (GWAS) of hematoma volume with the aim of identifying novel biological pathways involved in the pathophysiology of primary brain injury in ICH. We conducted a 2-stage (discovery and replication) case-only genome-wide association study in patients with ICH of European ancestry. We utilized the admission head computed tomography to calculate hematoma volume via semiautomated computer-assisted technique. After quality control and imputation, 7 million genetic variants were available for association testing with ICH volume, which was performed separately in lobar and nonlobar ICH cases using linear regression. Signals with P<5×10-8 were pursued in replication and tested for association with admission Glasgow coma scale and 3-month post-ICH dichotomized (0-2 versus 3-6) modified Rankin Scale using ordinal and logistic regression, respectively. The discovery phase included 394 ICH cases (228 lobar and 166 nonlobar) and identified 2 susceptibility loci: a genomic region on 22q13 encompassing PARVB (top single-nucleotide polymorphism rs9614326: β, 1.84; SE, 0.32; P=4.4×10-8) for lobar ICH volume and an intergenic region overlying numerous copy number variants on 17p12 (top single-nucleotide polymorphism rs11655160: β, 0.95; SE, 0.17; P=4.3×10-8) for nonlobar ICH volume. The replication included 240 ICH cases (71 lobar and 169 nonlobar) and corroborated the association for 17p12 (P=0.04; meta-analysis P=2.5×10-9; heterogeneity, P=0.16) but not for 22q13 (P=0.49). In multivariable analysis, rs11655160 was also associated with lower admission Glasgow coma scale (odds ratio, 0.17; P=0.004) and increased risk of poor 3-month modified Rankin Scale (odds ratio, 1.94; P=0.045). We identified 17p12 as a novel susceptibility risk locus for hematoma volume, clinical severity, and functional outcome in nonlobar ICH. Replication in other ethnicities and follow-up translational studies are needed to elucidate the mechanism mediating the observed association.


Localisation and characteristics of natriuretic peptide receptors in the gills of the Atlantic hagfish Myxine glutinosa (Agnatha).
Specific binding of iodinated natriuretic peptides 125I-ANP and 125I-CNP was examined in the gill of the Atlantic hagfish Myxine glutinosa by tissue section autoradiography, saturation and competition analysis of binding to membrane preparations, affinity cross-linking, followed by SDS-PAGE and guanylate cyclase assays. Autoradiographs showed specific, saturable binding on the respiratory lamellar epithelium. In vitro analysis of the binding sites demonstrated that 125I-ANP bound to two receptor sites with the same affinity (Kd = 15.4 +/- 1.6 pmol l-1; Bmax = 45.9 +/- 3.0 fmol mg-1 protein). 125I-CNP bound to high- and low-affinity receptor sites; variables for the high-affinity site (Kd = 12.9 +/- 4.7 pmol l-1; Bmax = 23.4 +/- 6.5 fmol mg-1 protein) did not differ from those for the 125I-ANP sites. The low-affinity site had an apparent Kd and Bmax of 380 +/- 80 pmol l-1 and 120 +/- 21 fmol mg-1 protein, respectively. All receptors had an apparent molecular mass of approximately 150 kDa, with no indication of a mammalian type NPR-C at a lower apparent molecular mass. 1 nmol l-1 unlabelled rANP and 20 and 30 nmol l-1 unlabelled pCNP and C-ANF, respectively, competed for 50% of 125I-ANP sites. 0.1 nmol l-1 rANP and pCNP and 8 nmol l-1 C-ANF competitively inhibited 50% of 125I-CNP binding. Both rANP and pCNP stimulated cyclic GMP production, although rANP was a more potent stimulator than was pCNP. C-ANF did not stimulate cyclic GMP production. These data suggest the existence of an ANP guanylate-cyclase-linked receptor similar to the mammalian NPR-A and an ANP/CNP receptor that may be similar to, although not structurally homologous with, the mammalian NPR-C clearance receptor.


Aortic Calcification Affects Noninvasive Estimates of Central Blood Pressure in Patients with Severe Chronic Kidney Disease.
Central blood pressure (BP) assessed noninvasively considerably underestimates true invasively measured aortic BP in chronic kidney disease (CKD) patients. The difference between the estimated and the true aortic BP increases with decreasing estimated glomerular filtration rates (eGFR). The present study investigated whether aortic calcification affects noninvasive estimates of central BP. Twenty-four patients with CKD stage 4-5 undergoing coronary angiography and an aortic computed tomography scan were included (63% males, age [mean ± SD ] 53 ± 11 years, and eGFR 9 ± 5 mL/min/1.73 m2). Invasive aortic BP was measured through the angiography catheter, while non-invasive central BP was obtained using radial artery tonometry with a SphygmoCor® device. The Agatston calcium score (CS) in the aorta was quantified on CT scans using the CS on CT scans. The invasive aortic systolic BP (SBP) was 152 ± 23 mm Hg, while the estimated central SBP was 133 ± 20 mm Hg. Ten patients had a CS of 0 in the aorta, while 14 patients had a CS >0 in the aorta. The estimated central SBP was lower than the invasive aortic SBP in patients with aortic calcification compared to patients without (mean difference 8 mm Hg, 95% CI 0.3-16; p = 0.04). The brachial SBP was lower than the aortic SBP in patients with aortic calcification compared to patients without (mean difference 10 mm Hg, 95% CI 2-19; p = 0.02). In patients with advanced CKD the presence of aortic calcification is associated with a higher difference between invasively measured central aortic BP and non-invasive estimates of central BP as compared to patients without calcifications.


Good oral health and diet.
An unhealthy diet has been implicated as risk factors for several chronic diseases that are known to be associated with oral diseases. Studies investigating the relationship between oral diseases and diet are limited. Therefore, this study was conducted to describe the relationship between healthy eating habits and oral health status. The dentistry has an important role in the diagnosis of oral diseases correlated with diet. Consistent nutrition guidelines are essential to improve health. A poor diet was significantly associated with increased odds of oral disease. Dietary advice for the prevention of oral diseases has to be a part of routine patient education practices. Inconsistencies in dietary advice may be linked to inadequate training of professionals. Literature suggests that the nutrition training of dentists and oral health training of dietitians and nutritionists is limited.


The XVP/ NAC003 protein associates with the plasma membrane through KR rich regions and translocates to the nucleus by changing phosphorylation status.
Membrane localized transcription factors play essential roles in various plant developmental processes. The XVP/NAC003 protein is a NAC domain transcription factor associated with the plasma membrane and involved in the TDIF-PXY signaling during vascular development. We report here the mechanisms of XVP membrane localization and its nuclear translocation. Using a transient transformation approach, we found that XVP is associated with the plasma membrane through positively charged KR-rich regions. Mutagenesis studies found that the threonine amino acid at position 354 (T354) is critical for XVP translocation to the nucleus. In particular, the threonine to alanine mutation (T354A) resulted in a partial nucleus localization, while threonine to aspartic acid (T354D) mutation showed no effect on protein localization, indicating that dephosphorylation at T354 may serve as a nucleus translocation signal. This research sheds new light on the nucleus partitioning of plasma membrane-associated transcription factors.


Annual climate variation modifies nitrogen induced carbon accumulation of Pinus sylvestris forests.
We report results from long-term simulated external nitrogen (N) input experiments in three northern Pinus sylvestris forests, two of moderately high and one of moderately low productivity, assessing effects on annual net primary production (NPP) of woody mass and its interannual variation in response to variability in weather conditions. A sigmoidal response of wood NPP to external N inputs was observed in the both higher and lower productivity stands, reaching a maximum of ~65% enhancement regardless of the native site productivity, saturating at an external N input of 4-5 g N·m-2 ·yr-1 . The rate of increase in wood NPP and the N response efficiency (REN , increase in wood NPP per external N input) were maximized at an external N input of ~3 g N·m-2 ·yr-1 , regardless of site productivity. The maximum REN was greater in the higher productivity than the lower productivity stand (~20 vs. ~14 g C/g N). The N-induced enhancement of wood NPP and its REN were, however, markedly contingent on climatic variables. In both of the higher and lower productivity stands, wood NPP increased with growing season precipitation (P), but only up to ~400 mm. The sensitivity of the response to P increased with increasing external N inputs. Increasing growing season temperature (T) somewhat increased the N-induced drought effect, whereas decreasing T reduced the drought effect. These responses of wood NPP infused a large temporal variation to REN , making the use of a fixed value unadvisable. Based on these results, we suggest that regional climate conditions and future climate scenarios should be considered when modeling carbon sequestration in response to N deposition in boreal P. sylvestris, and possibly other forests.


Integrating binary traits with quantitative phenotypes for association mapping of multivariate phenotypes.
Clinical binary end-point traits are often governed by quantitative precursors. Hence it may be a prudent strategy to analyze a clinical end-point trait by considering a multivariate phenotype vector, possibly including both quantitative and qualitative phenotypes. A major statistical challenge lies in integrating the constituent phenotypes into a reduced univariate phenotype for association analyses. We assess the performances of certain reduced phenotypes using analysis of variance and a model-free quantile-based approach. We find that analysis of variance is more powerful than the quantile-based approach in detecting association, particularly for rare variants. We also find that using a principal component of the quantitative phenotypes and the residual of a logistic regression of the binary phenotype on the quantitative phenotypes may be an optimal method for integrating a binary phenotype with quantitative phenotypes to define a reduced univariate phenotype.


An amino-terminal tetrapeptide specifies cotranslational degradation of beta-tubulin but not alpha-tubulin mRNAs.
The steady-state level of alpha- and beta-tubulin synthesis is autoregulated by a posttranscriptional mechanism that selectively alters alpha- and beta-tubulin mRNA levels in response to changes in the unassembled tubulin subunit concentration. For beta-tubulin mRNAs, previous efforts have shown that this is the result of a selective mRNA degradation mechanism which involves cotranslational recognition of the nascent amino-terminal beta-tubulin tetrapeptide as it emerges from the ribosome. Site-directed mutagenesis is now used to determine that the minimal sequence requirement for conferring the full range of beta-tubulin autoregulation is the amino-terminal tetrapeptide MR(E/D)I. Although tubulin-dependent changes in alpha-tubulin mRNA levels are shown to result from changes in cytoplasmic mRNA stability, transfection of wild-type and mutated alpha-tubulin genes reveals that alpha- and beta-tubulin mRNA degradation is not mediated through a common pathway. Not only does the amino-terminal alpha-tubulin tetrapeptide MREC fail to confer regulated mRNA degradation, neither wild-type alpha-tubulin transgenes nor an alpha-tubulin gene mutated to encode an amino-terminal MREI yields mRNAs that are autoregulated. Further, although slowing ribosome transit accelerates the autoregulated degradation of endogenous alpha- and beta-tubulin mRNAs, degradation of alpha-tubulin transgene mRNAs is not enhanced, and in one case, the mRNA is actually stabilized. We conclude that, despite similarities, alpha- and beta-tubulin mRNA destabilization pathways utilize divergent determinants to link RNA instability to tubulin subunit concentrations.


N-formyl-L-methionine deformylase activity in human leucocytes and platelets.
Extracts of human neutrophils, lymphocytes and platelets enzymically deformylate N-formyl-L-methionine. Enzyme activity is stimulated by Co2+, inhibited by bivalent-cation chelators and unaffected by inhibitors of serine, thiol and carboxyl proteinases. Leucocyte or platelet N-formylmethionine deformylase may be important in modulation of neutrophil responses to chemoattractant formylmethionyl peptides or similar compounds.