Changes in aberrations and retinal image quality due to tear film dynamics.
A reliable and objective method to measure aberration changes due to the tear film is essential in improving clinical assessment of the tear film and in vivo retinal imaging. The tear film of 11 subjects are studied by acquiring continuous wavefront measurements in real-time with a customized Shack-Hartmann wavefront sensor. The device has a high resolution lenslet array (190 mum) and a topographer unit with an infrared pupil illuminator (940 nm). A Fourier transform reconstructor algorithm [1] was used to estimate the eyes' wavefront aberrations from slope measurements. Increasing irregularities in the tear film produced observable wavefront variations. The temporal behavior of tear induced aberrations and retinal image quality was evaluated by the root mean squared (RMS) error of the residual wavefront and the volume modulation transfer function (MTF). Similar trends were observed from both metrics. Our analysis demonstrates the applicability of the SH wavefront sensor to assessing the dynamics of the human tear film.


Vibration exposure and biodynamic responses during whole-body vibration training.
Excessive, chronic whole-body vibration (WBV) has a number of negative side effects on the human body, including disorders of the skeletal, digestive, reproductive, visual, and vestibular systems. Whole-body vibration training (WBVT) is intentional exposure to WBV to increase leg muscle strength, bone mineral density, health-related quality of life, and decrease back pain. The purpose of this study was to quantitatively evaluate vibration exposure and biodynamic responses during typical WBVT regimens. Healthy men and women (N = 16) were recruited to perform slow, unloaded squats during WBVT (30 Hz; 4 mm(p-p)), during which knee flexion angle (KA), mechanical impedance, head acceleration (Ha(rms)), and estimated vibration dose value (eVDV) were measured. WBVT was repeated using two forms of vibration: 1) vertical forces to both feet simultaneously (VV), and 2) upward forces to only one foot at a time (RV). Mechanical impedance varied inversely with KA during RV (effect size, eta(p)(2): 0.668, P < 0.01) and VV (eta(p)(2): 0.533, P < 0.05). Ha(rms) varied with KA (eta(p)(2): 0.686, P < 0.01) and is greater during VV than during RV at all KA (P < 0.01). The effect of KA on Ha(rms) is different for RV and VV (eta(p)(2): 0.567, P < 0.05). The eVDV associated with typical RV and VV training regimens (30 Hz, 4 mm(p-p), 10 min.d(-1)) exceeds the recommended daily vibration exposure as defined by ISO 2631-1 (P < 0.01). ISO standards indicate that 10 min.d(-1) WBVT is potentially harmful to the human body; the risk of adverse health effects may be lower during RV than VV and at half-squats rather than full-squats or upright stance. More research is needed to explore the long-term health hazards of WBVT.


Partial adenosine deaminase deficiency: another family from southern Africa.
Adenosine deaminase (ADA) from a partially ADA-deficient Xhosa man has been characterized. This is only the second such case described in southern Africa, the previous one being a Kalahari San ("Bushman"). Red blood cell ADA levels were found to be only 6-9% of normal whereas his white cell ADA levels were much higher at 30% of normal. The stability of the enzyme at 57 degrees C was shown to be greatly decreased indicating a mutation resulting in an enzyme with decreased stability in vivo. The Michaelis constant (Km) for adenosine was found to be normal. Deoxy-ATP levels in the red cells were elevated 2- to 3-times above normal, although this appears to be of no immunological consequence. Starch gel electrophoresis of red cell ADA from family members of the index case, in conjunction with red cell ADA activity levels, suggested that both parents carried a gene for partial ADA deficiency. Isoelectric focusing studies suggested that the two parental partial ADA-deficiency genes were not the same. Electrophoretic studies also revealed that another rare allele of ADA, possibly ADA*5, was segregating within the same family although this event appears to be unconnected with the ADA partial deficiency. A Xhosa population sample was assayed for red cell ADA activity. The results suggested a frequency of 0.015 +/- 0.010 for ADA partial-deficiency alleles, although the number of different alleles involved is not known.


Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci by PCR.
A PCR assay that allows simultaneous detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci (Enterococcus faecium, E. faecalis, E. gallinarum, and E. casseliflavus) was developed. This assay was based on specific amplification of internal fragments of genes encoding D-alanine:D-alanine ligases and related glycopeptide resistance proteins. The specificity of the assay was tested on 5 well-characterized glycopeptide-resistant strains and on 15 susceptible enterococcal type strains. Clinical isolates of enterococci that could not be identified to the species level by conventional methods were identified by the PCR test. This assay offers a specific and rapid alternative to antibiotic susceptibility tests, in particular for detection of low-level vancomycin resistance.


Structure of the Dimerization Interface in the Mature HIV-1 Capsid Protein Lattice from Solid State NMR of Tubular Assemblies.
The HIV-1 capsid protein (CA) forms the capsid shell that encloses RNA within a mature HIV-1 virion. Previous studies by electron microscopy have shown that the capsid shell is primarily a triangular lattice of CA hexamers, with variable curvature that destroys the ideal symmetry of a planar lattice. The mature CA lattice depends on CA dimerization, which occurs through interactions between helix 9 segments of the C-terminal domain (CTD) of CA. Several high-resolution structures of the CTD-CTD dimerization interface have been reported, based on X-ray crystallography and multidimensional solution nuclear magnetic resonance (NMR), with significant differences in amino acid side chain conformations and helix 9-helix 9 orientations. In a structural model for tubular CA assemblies based on cryogenic electron microscopy (cryoEM) [Zhao et al. Nature, 2013, 497, 643-646], the dimerization interface is substantially disordered. The dimerization interface structure in noncrystalline CA assemblies and the extent to which this interface is structurally ordered within a curved lattice have therefore been unclear. Here we describe solid state NMR measurements on the dimerization interface in tubular CA assemblies, which contain the curved triangular lattice of a mature virion, including quantitative measurements of intermolecular and intramolecular distances using dipolar recoupling techniques, solid state NMR chemical shifts, and long-range side chain-side chain contacts. When combined with restraints on the distance and orientation between helix 9 segments from the cryoEM study, the solid state NMR data lead to a unique high-resolution structure for the dimerization interface in the noncrystalline lattice of CA tubes. These results demonstrate that CA lattice curvature is not dependent on disorder or variability in the dimerization interface. This work also demonstrates the feasibility of local structure determination within large noncrystalline assemblies formed by high-molecular-weight proteins, using modern solid state NMR methods.


Concerted nonsyntenic allelic losses in hyperploid hepatocellular carcinoma as determined by a high-resolution allelotype.
Although the occurrence of loss of genetic material in hepatocellular carcinoma (HCC) has been documented both by cytogenetic analysis and by monitoring of allelic losses, a global overview of the extent and frequency of deletion occurring throughout the genome is not yet available. To contribute to this information, DNAs extracted from flow-sorted aneuploid nuclei from HCC and matched normal DNAs were typed for 275 microsatellite loci that were distributed along the autosomes. An average of 190 (69%) informative loci per case were generated on 48 HCC. Complete loss of heterozygozity in the tumor DNA was observed for 15.6% of the typed loci. The chromosome segments that were most frequently affected by deletion were: 8p (60%), 17p (48%), 1p (44%), 4q (42%), 16p (40%), 16q (39%), 6q (35%), 9p (30%), and 13q (29%). On average, 8 of the 39 chromosome segments studied per tumor carried at least one locus that demonstrated loss of heterozygosity (ie., the fractional allelic loss was 0.21). Groups of concerted nonsyntenic losses were observed for 16p and 1p and for 16p and 4q. The location of putative tumor suppressor genes on the most frequently deleted regions was confirmed and, in some cases, refined.


Potassium currents and their modulation by muscarine and substance P in neuronal cultures from adult guinea pig celiac ganglia.
1. Intracellular microelectrode and whole-cell patch-clamp recordings were obtained from adult guinea pig celiac ganglion neurons grown in tissue culture for 7-14 days. Over 90% of neurons showed phasic-type action-potential discharge with the use of either type of recording electrode; they stained immunohistochemically for catecholamines, tyrosine hydroxylase, and neuropeptide Y. Input resistance (140 M omega) and action-potential amplitude (103 mV) were significantly greater with whole-cell than with microelectrode recordings, but other passive electrical properties were similar. 2. Five potassium currents were characterized: an apamin-sensitive after hyperpolarizing current (IAHP), an apamin and tetraethylammonium-insensitive slow IAHP, an M-like current, a transient outward IA current, and a delayed rectifier IK current. A hyperpolarization-activated cationic Ih current was also present. The first three currents were not observed with whole-cell recordings. 3. Cadmium (200 microM), cobalt (1 mM), lanthanum (30 microM), or a low calcium/high magnesium solution blocked both IAHPS and the M-like current; barium (1 mM) also blocked these currents. 4. Kinetics of the M-like current were best described by a double exponential fit to deactivating tail currents with time constants of 50 and 390 ms at -50 mV. The apamin-sensitive and slow IAHP decayed exponentially with time constants of 145 ms and 3.5 s, respectively. There was no correlation between occurrence of M-like current (95% of neurons) and slow IAHP (40% of neurons), nor any correlation between magnitude of M-like current and IAHP in those cells exhibiting both currents. 5. Muscarine and substance P (SP) caused depolarizations or inward currents (under voltage clamp) at the resting potential (-55 mV) associated with a decreased membrane conductance. The slow IAHP and the M-like current, but not the apamin-sensitive IAHP nor the IA, were blocked by muscarine and SP (IC50 3 microM and 100 nM, respectively). Muscarine and SP also decreased a "leak" potassium current. 6. We conclude that celiac neurons express two calcium-dependent IAHP currents and a calcium-dependent M-current; these are seen by fine-tipped intracellular microelectrodes but not by whole-cell patch electrodes. These currents are not required for spike frequency accommodation. Muscarine and SP reduce these currents, as well as voltage-independent leakage potassium current.


Ultrashort laser pulse characterization using modified spectrum auto-interferometric correlation (MOSAIC).
Sensitive, real-time chirp and spectral phase diagnostics along with full field reconstruction of femtosecond laser pulses are performed using a single rapid-scan interferometric autocorrelator. Through the use of phase retrieval error maps, ambiguities in pulse retrievals based on the pulse spectrum and various forms of MOSAIC traces are discussed. We show second-order autocorrelations can introduce significantly different amounts of chirp depending on the implementation. Examples are presented that illustrate the sensitivity and fidelity of the scheme even with low signal-to-noise.


PD-1/PD-L1 Targeting in Breast Cancer: The First Clinical Evidences Are Emerging. A Literature Review.
Recently, the development of immunotherapy through the immune checkpoint blockade led to long-lasting responses in several types of cancers that are refractory to conventional treatments, such as melanoma or non-small cell lung cancer. Immunotherapy has also demonstrated significant improvements in various other types of cancers. However, breast cancer remains one of the tumors that have not experienced the explosion of immunotherapy yet. Indeed, breast cancer was traditionally considered as being weakly immunogenic with a lower mutational load compared to other tumor types. In the last few years, anti-PD1/PD-L1 (Programmed death-ligand 1) agents have been evaluated in breast cancer, particularly in the triple negative subtype, with promising results observed when delivered as monotherapy or in combination with conventional treatments. In this review, we will report the results of the most recent studies evaluating immune checkpoint inhibitors in breast cancer. In addition, we will discuss the concomitant development of possible biomarkers, which is required for improving the selection of patients with the highest probability of benefiting from these agents.


Evaluation of the antioxidant and melanogenesis inhibitory properties of pracparatum mungo (lu-do huang).
Pracparatum mungo (Lu-Do Huang) is a traditional Chinese functional medicine made from the natural fermentation of mung bean (Lǜ Dòu) mixed with other Chinese medicines. It has been recognized as having liver protecting and detoxifying effects. As mung beans have been verified to possess anti-inflammatory, antioxidant, antipyretic, and whitening actions, the present research utilized the in vitro, ex vivo, and in vivo experimental models to investigate the antioxidant and melanin inhibiting effects of P. mungo on the skin. The in vitro experiment revealed that P. mungo methanol extract (PMME) and P. mungo ethanol extract (PMEE) possess the capacity to clear α,α-diphenyl-2-picrylhydrazyl (DPPH) radicals and inhibit tyrosinase activity. The ex vivo experiment indicated that PMEE can promote the growth of MDCK cells and increase the enzymatic activities of superoxide dismutase (SOD) and catalase in MDCK cells. On the other hand, PMME and PMEE can suppress the proliferation of A375 cells, and PMEE can reduce the enzymatic activities of SOD and catalase in A375 cells. The in vivo results showed that P. mungo can enhance the enzymatic performance of SOD, Catalase, and glutathione peroxidase (GPx) in the liver. The results also showed that P. mungo has antioxidant characteristics and can inhibit tyrosinase activity, thereby promoting the growth of skin tissues and suppressing the proliferation of A375 cells, and thus enhancing the effects that the antioxidant enzymatic performance has on the liver. These results can be applied in the development of tyrosinase inhibitors or antioxidants used for the inhibition of melanin biosynthesis or for auto-oxidation in further industrial applications, particularly those relating to functional food or cosmetic compositions.


A randomized, blinded, comparative trial of one pepsin-digested and two whole IgG antivenoms for Bothrops snake bites in Uraba, Colombia. The Regional Group on Antivenom Therapy Research (REGATHER).
The therapeutic efficacy and the incidence of early antivenom reactions (EARs) were compared in a clinical trial performed in 79 patients bitten by Bothrops sp. in Urabá, Colombia. Patients were randomized into three groups according to the antivenom administered: A (n = 30, Butantan polyspecific, pepsin-digested Bothrops antivenom); B (n = 27, Butantan polyspecific, whole IgG Bothrops antivenom); and C (n = 22, Colombian commercial, monovalent, whole IgG Bothrops antivenom). The groups were comparable in all clinical and epidemiologic aspects; 33 patients had mild, 22 moderate, and 24 severe envenoming. At the doses used (two, four, and six vials [10 ml/vial] for mild, moderate, and severe envenomings, respectively) there were no differences between the antivenoms in restoring normal hemostatic parameters within 24 hr. The evolution of local envenoming was comparable in the three groups. Serum venom/antivenom kinetics determined by ELISA showed a complete clearance of venom levels 1 hr after treatment in mild/moderate envenomings. In severe cases, venom levels remained detectable up to 24 hr and recurrence of antigenemia was observed in some cases. Antivenom concentrations remained at high levels up to 24 hr of treatment. The incidence of EARs was significantly different in the groups: A (36.7%), B (11.1.%), and C (81.8%). There were no life-threatening anaphylactic reactions. We conclude that the efficacy of the three antivenoms was similar in neutralizing human Bothrops envenomings and that the production of whole IgG antivenoms by caprylic acid fractionation is a good alternative for reducing the incidence of EARs.


Interaction of plasmin with endothelial cells.
Interaction of human plasmin with a monolayer culture of mini-pig aortic endothelial cells was studied by using the 125I-labelled enzyme. The binding of plasmin was time- and concentration-dependent. Equilibrium between bound and free enzyme was obtained within 90s, and Scatchard analysis indicated a high- and a low-affinity population of binding sites of approx. 1.24 X 10(4) sites/cell having a Kd of 1.4 X 10(-9) M and 7.2 X 10(4) sites/cell with a Kd of 2 X 10(-8) M respectively. Plasmin, bound to cell, was spontaneously released within 2 min, suggesting a rapid equilibrium. Chemical modification of the enzyme with phenylmethanesulphonyl fluoride or pyridoxal 5'-phosphate revealed that neither the active centre nor the heparin-binding site of plasmin was involved in the interaction with the endothelial cell. In terms of endothelial-cell receptors, the binding sites of cells for plasmin and thrombin were different: the two enzymes did not compete with each other, and the pretreatment of cells with neuraminidase or chondroitin ABC lyase resulted in a 50% decrease of thrombin or plasmin binding respectively. Arachidonic acid incorporated into phospholipids of the cell was released by plasmin, but a change in the rate of prostacyclin formation was not measurable. The interaction of plasmin with endothelial cells seems to be specific in the fibrinolytic system, since plasminogen did not bind to these cells under similar conditions.


Comparison of fat oxidation during exercise in lean and obese pubertal boys: clinical implications.
To examine fat oxidation rates during exercise in lean and obese pubescent children. A graded leg cycle ergometry test was performed by two groups of pubescent boys (13 lean: mean (SD) age 12.0 (0.5) years, body mass index (BMI) 18.56 (1.12) kg/m(2); 17 obese: mean (SD) age 12.1 (0.1) years, BMI 26.68 (3.37) kg/m(2); p<0.001). The first step of the test was fixed at 30 W and power was gradually increased by 20 W every 3.5 min. The mean ventilatory gas measurement was obtained during the last 30 s of each step for calculation of fat oxidation rate vs exercise intensity. At low intensity (0-30% of peak oxygen consumption) when fat-free mass is considered, the fat oxidation rate was identical for the two groups. At higher intensities (40%, 50% and 60% of peak oxygen consumption) the fat oxidation rate was significantly higher in lean boys than in obese boys. These results confirm that obese pubertal boys have fat-free mass decreased capacities to use fat during moderate exercise. The findings suggest that obese boys need to practise physical activity at a lower intensity than healthy boys to enhance lipolysis and diminish adipose tissue and the consequences of obesity.


A comparison of cluster and systematic sampling methods for measuring crude mortality.
To compare the results of two different survey sampling techniques (cluster and systematic) used to measure retrospective mortality on the same population at about the same time. Immediately following a cluster survey to assess mortality retrospectively in a town in North Darfur, Sudan in 2005, we conducted a systematic survey on the same population and again measured mortality retrospectively. This was only possible because the geographical layout of the town, and the availability of a good previous estimate of the population size and distribution, were conducive to the systematic survey design. Both the cluster and the systematic survey methods gave similar results below the emergency threshold for crude mortality (0.80 versus 0.77 per 10,000/day, respectively). The results for mortality in children under 5 years old (U5MR) were different (1.16 versus 0.71 per 10,000/day), although this difference was not statistically significant. The 95% confidence intervals were wider in each case for the cluster survey, especially for the U5MR (0.15-2.18 for the cluster versus 0.09-1.33 for the systematic survey). Both methods gave similar age and sex distributions. The systematic survey, however, allowed for an estimate of the town's population size, and a smaller sample could have been used. This study was conducted in a purely operational, rather than a research context. A research study into alternative methods for measuring retrospective mortality in areas with mortality significantly above the emergency threshold is needed, and is planned for 2006.


The olfactory secretome varies according to season in female sheep and goat.
Small ungulates (sheep and goat) display a seasonal breeding, characterised by two successive periods, sexual activity (SA) and sexual rest (SR). Odours emitted by a sexually active male can reactivate the ovulatory cycle of anoestrus females. The plasticity of the olfactory system under these hormonal changes has never been explored at the peripheral level of odours reception. As it was shown in pig that the olfactory secretome (proteins secreted in the nasal mucus) could be modified under hormonal control, we monitored its composition in females of both species through several reproductive seasons, thanks to a non-invasive sampling of olfactory mucus. For this purpose, two-dimensional gel electrophoresis (2D-E), western-blot with specific antibodies, MALDI-TOF and high-resolution (nano-LC-MS/MS) mass spectrometry, RACE-PCR and molecular modelling were used. In both species the olfactory secretome is composed of isoforms of OBP-like proteins, generated by post-translational modifications, as phosphorylation, N-glycosylation and O-GlcNAcylation. Important changes were observed in the olfactory secretome between the sexual rest and the sexual activity periods, characterised in ewe by the specific expression of SAL-like proteins and the emergence of OBPs O-GlcNAcylation. In goat, the differences between SA and SR did not come from new proteins expression, but from different post-translational modifications, the main difference between the SA and SR secretome being the number of isoforms of each protein. Proteomics data are available via ProteomeXchange with identifier PXD014833. Despite common behaviour, seasonal breeding, and genetic resources, the two species seem to adapt their olfactory equipment in SA by different modalities: the variation of olfactory secretome in ewe could correspond to a specialization to detect male odours only in SA, whereas in goat the stability of the olfactory secretome could indicate a constant capacity of odours detection suggesting that the hallmark of SA in goat might be the emission of specific odours by the sexually active male. In both species, the olfactory secretome is a phenotype reflecting the physiological status of females, and could be used by breeders to monitor their receptivity to the male effect.


Efficacy of full-fat milk and diluted lemon juice in reducing infra-cardiac activity of (99m)Tc sestamibi during myocardial perfusion imaging.
When using (99m)Tc sestamibi for myocardial perfusion imaging, increased splanchnic activity creates a problem in the visual and quantitative interpretation of the inferior and infero-septal walls of the left ventricle. We sought to determine whether the administration of diluted lemon juice or full-fat milk would be effective in reducing interfering infra-cardiac activity and therefore result in an improvement in image quality. We compared the administration of full-fat milk and diluted lemon juice to a control group that had no intervention. The study was carried out prospectively. All patients referred to our institution for myocardial perfusion imaging from November 2009 to May 2012 were invited to be enrolled in the study. A total of 630 patients were randomised into three groups. Group 0 (G0), 246 patients, were given diluted lemon juice, group 1 (G1), 313 patients, were given full-fat milk, and group 2 (G2), 71 patients, had no intervention (control group). A routine two-day protocol was used and the patients were given the same intervention on both days. Raw data of both the stress and rest images were visually assessed for the presence of infra-cardiac activity, and quantitative grading of the relative intensity of myocardial activity to infra-cardiac activity was determined. The physicians were blinded to the intervention received and the data were reviewed simultaneously. The overall incidence of interfering infra-cardiac activity at stress was 84.1, 84.5 and 96.6% in G0, G1 and G2, respectively (p = 0.005). At rest it was 91.7, 90.1 and 100% in G0, G1 and G2, respectively (p = 0.0063). The visual and quantitative results favoured both milk and lemon juice in reducing the amount of interfering infra-cardiac activity versus no intervention. The administration of milk or lemon juice resulted in a significant decrease in the intensity of infra-cardiac activity compared to the control group. This reduction in intensity was even more significant in the milk group for patients assessed during rest myocardial perfusion imaging.


A molecular-modeling toolbox aimed at bridging the gap between medicinal chemistry and computational sciences.
In the current era of high-throughput drug discovery and development, molecular modeling has become an indispensable tool for identifying, optimizing and prioritizing small-molecule drug candidates. The required background in computational chemistry and the knowledge of how to handle the complex underlying protocols, however, might keep medicinal chemists from routinely using in silico technologies. Our objective is to encourage those researchers to exploit existing modeling technologies more frequently through easy-to-use graphical user interfaces. In this account, we present two innovative tools (which we are prepared to share with academic institutions) facilitating computational tasks commonly utilized in drug discovery and development: (1) the VirtualDesignLab estimates the binding affinity of small molecules by simulating and quantifying their binding to the three-dimensional structure of a target protein; and (2) the MD Client launches molecular dynamics simulations aimed at exploring the time-dependent stability of ligand-protein complexes and provides residue-based interaction energies. This allows medicinal chemists to identify sites of potential improvement in their candidate molecule. As a case study, we present the application of our tools towards the design of novel antagonists for the FimH adhesin.


The spectacle of the ball python (Python regius): a morphological description.
A detailed morphological description of the spectacle of the ball python (Python regius) is provided. The eyes of 21 snakes were examined by light microscopy and/or transmission electron microscopy. Additionally, eyes of nine live snakes were examined using optical coherence tomography (OCT) and Scheimpflug scanning (Pentacam). The spectacle consists of three layers: outer epithelium, stroma and inner epithelium. The outer epithelium is made up of flat basal cells overlaid by keratin, the stroma consists of organized layers of collagen fibrils with interweaving nerve fibers and blood vessels, and the inner epithelium holds squamous cells containing vesicles and microvilli. At the rim of the spectacle, there is a transition zone, where the spectacle merges with the epidermis and dermis of the periocular scales. This zone is characterized by a greater height of the basal cells of the outer epithelium and a less orderly organization of the stroma compared with the spectacle proper. The thickness of the spectacle was uniform throughout. It averaged 96 ± 10 µm in histological specimens and 108 ± 13 µm using OCT. The subspectacular space was extremely narrow in the live snakes; however, the space was visible at the periphery of the spectacle with OCT.


Recognition and catabolism of synthetic heterotrimeric collagen peptides by matrix metalloproteinases.
The general consensus is that interstitial collagens are digested by collagenases and denatured collagen by gelatinases, although processing of fibrillar and acetic-acid-soluble collagen by gelatinase A has also been reported. One of the main difficulties in studying the mechanism of action of these matrix metalloproteinases (MMPs) derives from the physicochemical properties of the natural triple-helical collagen, which makes it difficult to handle. Synthetic heterotrimeric collagenous peptides that contain the collagenase cleavage site of human collagen type I and differ in the thermal stability of the triple-helical fold were used to mimic natural collagen and gelatin, respectively. Results from digestion of these substrates by fibroblast and neutrophil collagenases (MMP-1 and MMP-8), as well as by gelatinase A (MMP-2), confirmed that the two classes of enzymes operate within the context of strong conformational dependency of the substrates. It was also found that gelatinases and collagenases exhibit two distinct proteolytic mechanisms: gelatinase digests the gelatin-like heterotrimer rapidly in individual steps with intermediate releases of partially processed substrate into the medium, whereas collagenases degrade the triple-helical heterotrimer by trapping it until scission through all three alpha chains is achieved. The results confirm the usefulness of synthetic heterotrimeric collagenous peptides in the folded and unfolded state as mimics of the natural substrates collagen and gelatin, respectively, to gain a better a insight into the proteolytic mechanisms of matrix metalloproteinases.


Methamphetamine Use and Its Correlates among Individuals with Opioid Use Disorder in a Midwestern U.S. City.
Background: U.S. is experiencing a surging trend of methamphetamine use among individuals who use opioids. More research is needed to characterize this emerging "twin epidemic." Objectives: The study aims to identify social and behavioral characteristics associated with methamphetamine use among individuals with opioid use disorder (OUD) in the Dayton, Ohio, area, an epicenter of the opioid crisis and an emerging frontier of methamphetamine epidemic. Methods: 357 adult individuals with current OUD were recruited using targeted and respondent-driven sampling. Structured interviews collected information on social and drug use characteristics. Multivariable Logistic Regression was used to identify characteristics associated with the past 6-month use of methamphetamine. Results: 49.7% were female, and 88.8% were non-Hispanic whites. 55.6% used methamphetamine in the past 6-months, and 84.9% reported first use of methamphetamine after initiation of illicit opioids. Methamphetamine use was associated with homelessness (aOR = 2.46, p = .0001), lifetime history of diverted pharmaceutical stimulant use (aOR = 2.97, p < .001), injection route of heroin/fentanyl use (aOR = 1.89, p = .03), preference for fentanyl over heroin (aOR = 1.82, p = .048), lifetime history of extended-release injectable naltrexone (Vivitrol)-based treatment (aOR = 2.89, p = .003), and more frequent marijuana use (aOR = 1.26, p = .04). Discussion: The findings point to the complexity of motivational and behavioral pathways associated with methamphetamine and opioid co-use, ranging from self-treatment and substitution behaviors, attempts to endure homelessness, and greater risk taking to experience euphoria. More research is needed to understand the causal relationships and the association between methamphetamine and Vivitrol use. Public health responses to the opioid crisis need to be urgently expanded to address the growing epidemic of methamphetamine use.


State aggregation and population dynamics in linear systems.
We consider complex systems that are composed of many interacting elements, evolving under some dynamics. We are interested in characterizing the ways in which these elements may be grouped into higher-level, macroscopic states in a way that is compatible with those dynamics. Such groupings may then be thought of as naturally emergent properties of the system. We formalize this idea and, in the case that the dynamics are linear, prove necessary and sufficient conditions for this to happen. In cases where there is an underlying symmetry among the components of the system, group theory may be used to provide a strong sufficient condition. These observations are illustrated with some artificial life examples.


Intracellular Ca2+ buffers disrupt muscarinic suppression of Ca2+ current and M current in rat sympathetic neurons.
The role of intracellular Ca2+ concentration ([Ca2+]i) in the muscarinic suppression of Ca2+ current and M-type K+ current has been investigated in isolated rat sympathetic neurons using the whole-cell patch-clamp technique and fura-2 fluorescence measurements. Muscarinic stimulation suppressed currents without raising [Ca2+]i. Nonetheless, intracellular bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetate (BAPTA) (11-12 mM), a Ca2+ chelator, reduced Ca2(+)-current suppression from 82 to 15%. For the latter, we explain the BAPTA action by a requirement for a certain minimum [Ca2+]i for continued operation of the pathway coupling muscarinic receptors to M-type K+ channels. The pathway coupling muscarinic receptors to Ca channels also showed some dependence on [Ca2+]i, but there may also be a blocking action of BAPTA that is independent of Ca2+ chelation.


A new definition of drowning: towards documentation and prevention of a global public health problem.
Drowning is a major global public health problem. Effective prevention of drowning requires programmes and policies that address known risk factors throughout the world. Surveillance, however, has been hampered by the lack of a uniform and internationally accepted definition that permits all relevant cases to be counted. To develop a new definition, an international consensus procedure was conducted. Experts in clinical medicine, injury epidemiology, prevention and rescue from all over the world participated in a series of "electronic" discussions and face-to-face workshops. The suitability of previous definitions and the major requirements of a new definition were intensely debated. The consensus was that the new definition should include both cases of fatal and nonfatal drowning. After considerable dialogue and debate, the following definition was adopted: "Drowning is the process of experiencing respiratory impairment from submersion/immersion in liquid." Drowning outcomes should be classified as: death, morbidity, and no morbidity. There was also consensus that the terms wet, dry, active, passive, silent, and secondary drowning should no longer be used. Thus a simple, comprehensive, and internationally accepted definition of drowning has been developed. Its use should support future activities in drowning surveillance worldwide, and lead to more reliable and comprehensive epidemiological information on this global, and frequently preventable, public health problem.


Beta-lipotropin-beta-endorphin response to low-dose ovine corticotropin releasing factor in endogenous depression. Preliminary studies.
Studies in depression using a maximal stimulatory dose of corticotropin releasing factor have concluded that elevated resting cortisol levels in depressed patients exert a negative feedback effect on the corticotroph, resulting in a decreased corticotropin response. In this preliminary report, we examine the effects of a submaximal dose of corticotropin releasing factor on the release of another corticotroph secretory product, beta-lipotropin-beta-endorphin. We observed a decreased beta-lipotropin-beta-endorphin response in depressed subjects, but a normal adrenal cortisol response. Although the total beta-lipotropin-beta-endorphin response was decreased, the initial secretory response did not differ between patients and normal controls. Rather, the patients appeared to turn off secretion faster. This rapid shutoff was seen in all patients regardless of resting cortisol levels, suggesting that resting cortisol levels alone do not explain the decreased response seen in depressed patients.


Antinociceptive action of 2-(4-bromobenzoyl)-3-methyl-4,6-dimethoxy benzofuran, a novel xanthoxyline derivative on chemical and thermal models of nociception in mice.
The antinociceptive effect of the novel xanthoxyline derivative 2-(4-bromobenzoyl)-3-methyl-4-6-dimethoxy benzofuran) (BMDB), given i.p., p.o., s.c., subplantarly, intrathecally or by i.c.v. routes was assessed in five models of chemical and thermal nociception in mice, namely acetic acid-induced abdominal constriction, formalin and capsaicin-induced licking, hot-plate and tail-flick tests. BMDB given by i.p., s.c., subplantarly or by i.c.v. routes elicited dose-related and long-lasting (4 hr) antinociception, but had no significant effect by p.o. route. At the ID50 level, this compound was about 15- to 100-fold more potent than aspirin and acetaminophen, but it was about 2- to 50-fold less potent than morphine. Its analgesic action was not influenced by naloxone, L-arginine, antagonist of alpha-1 adrenoceptor, prazosin, tau -aminobutyric acid (B) antagonist, phaclofen, after adrenalectomy, but was reversed in part by p-chlorophenylalanine methyl ester. Its analgesic action was not secondary to an anti-inflammatory effect, or was it associated with nonspecific effect such as muscle relaxant or sedative actions of animals. We conclude that BMDB produces dose-dependent spinal and supraspinal antinociception as evaluated in several algesic models in mice, including the neurogenic nociception responses induced by formalin and capsaicin. Its antinociceptive effect was insensitive to naloxone, suggesting the lack of involvement of endogenous opioid, was not modulated by adrenal glands and does not involve interaction with the L-arginine-nitric oxide pathway, activation of alpha-1 adrenoceptors or tau-aminobutyric acidB receptors, but requires, at least in part, the serotoninergic pathway.


Coordinate regulation of endothelin and adrenomedullin secretion by oxidative stress in endothelial cells.
To elucidate the significance of oxidative stress in the modulation of endothelial functions, we examined the effects of H(2)O(2) on the expression of two endothelium-derived vasoactive peptides, endothelin (ET) and adrenomedullin (Am), and their interaction. H(2)O(2) dose dependently suppressed ET secretion and ET-1 mRNA expression in bovine carotid endothelial cells (ECs). Menadion sodium bisulfate, a redox cycling drug, also decreased ET secretion in a dose-dependent manner. Catalase, a H(2)O(2) reductase, and dl-alpha-tocopherol (vitamin E) significantly inhibited H(2)O(2)-induced suppression of ET secretion. Downregulation of ET-1 mRNA under oxidative stress was regulated at the transcriptional level. In contrast, H(2)O(2) increased Am secretion (and its mRNA expression) accompanied by the augmentation of cAMP production. Am, as well as 8-bromo-cAMP and forskolin decreased ET secretion in a dose-dependent fashion. Furthermore, an anti-Am monoclonal antibody that we developed abolished H(2)O(2)-induced suppression of ET secretion at 6-24 h after the addition of H(2)O(2). H(2)O(2) increased the intracellular Ca(2+) concentration ([Ca(2+)](i)). Moreover, treatment with ionomycin, a Ca(2+) ionophore, and thapsigargin, an inhibitor of endoplasmic reticulum ATPase, decreased ET secretion dose dependently for 3 h. These results suggest that the production of ET was decreased via activation of the Am-cAMP pathway and by the elevation of [Ca(2+)](i) under oxidative stress. These findings elucidate the coordinate expression of two local vascular hormones, ET and Am, under oxidative stress, which may protect against vascular diseases.


Interaction between the cytodomains of the alpha 3 and beta 1 integrin subunits regulates remodelling of adhesion complexes on laminin.
The first step of laminin 1-induced signal transduction is initiated by the formation of alpha 6 beta 1 integrin-specific adhesion complexes. In contrast, on other laminin isoforms the adhesion complexes are alpha 3 beta 1 integrin-specific due to a transdominant regulation of the alpha 6 beta 1 integrin by the alpha 3 beta 1 integrin. To determine the mechanism of this regulation, peptides representing the cytoplasmic domain of the alpha 3 or alpha 6 integrin subunits were microinjected together with recombinant enhanced green fluorescence protein into live fibroblasts. Microinjection of the alpha 3 integrin peptide to laminin 1-adherent cells displaying alpha 6 beta 1 integrin-specific adhesion complexes resulted in the disengagement of the alpha 6 beta 1 integrin, while microinjection of green fluorescence protein alone or in combination with the alpha 6 integrin cytodomain had no effect. Further surface plasmon resonance studies revealed that the cytodomain of the beta 1 integrin subunit interacts with low affinity with the cytoplasmic tail of the alpha 3 integrin subunit, but not with that of several other alpha subunits including alpha 6. These results imply that the cytoplasmic tails of the integrin alpha subunits play a critical role in the regulation of integrin-induced signal transduction. In particular, the intracellular tail of the alpha 3 integrin subunit controls the formation of adhesion complexes in cells adhering to laminins.


[Breast cancer patient satisfaction with immediate two-stage implant-based breast reconstruction].
For most breast cancer patients in the Czech Republic, breast reconstruction is available only in a delayed manner. In the Masaryk Memorial Cancer Institute (MMCI), suitable candidates are offered immediate breast reconstruction using tissue expander with later exchange to a permanent silicone implant. The aim of this study was to assess patient satisfaction with this type of reconstruction. Sixty-two women who had undergone surgery at the MMCI from 2007 through 2013 were sent a simple questionnaire developed by our working team. Fifty-seven patients completed the questionnaire. The data were evaluated by description methods and statistical tests. Patient response was 92%. The absolute majority of patients (56/57) would opt for this method again. The vast majority of patients (48/57) are generally satisfied with their reconstruction. Most women (8/14) younger than 50 years after the unilateral surgery would prefer synchronous contralateral prophylactic mastectomy and bilateral reconstruction if they could choose again. After bilateral surgery, reconstructed breasts are more often regarded as a part of the patients body. Dressed women rate their look substantially better than when they are undressed. As for self-esteem, these women are feeling excellent or good. Their psychosocial well-being in common situations is predominantly excellent. Their sexual well-being is significantly worse, and almost half of these women indicate occasional pain in their reconstructed breasts. The patients emphasize the need for appropriate information before the surgery. Immediate two-stage implant-based breast reconstruction is a suitable option for some breast cancer patients. With regard to the less natural cosmetic result and feeling of the implant-reconstructed breast, appropriate selection of women for this type of surgery is necessary and potential candidates must be thoroughly informed in the preoperative setting.


Titanium Dioxide Nanoparticles Trigger Loss of Function and Perturbation of Mitochondrial Dynamics in Primary Hepatocytes.
Titanium dioxide (TiO2) nanoparticles are one of the most highly manufactured and employed nanomaterials in the world with applications in copious industrial and consumer products. The liver is a major accumulation site for many nanoparticles, including TiO2, directly through intentional exposure or indirectly through unintentional ingestion via water, food or animals and increased environmental contamination. Growing concerns over the current usage of TiO2 coupled with the lack of mechanistic understanding of its potential health risk is the motivation for this study. Here we determined the toxic effect of three different TiO2 nanoparticles (commercially available rutile, anatase and P25) on primary rat hepatocytes. Specifically, we evaluated events related to hepatocyte functions and mitochondrial dynamics: (1) urea and albumin synthesis using colorimetric and ELISA assays, respectively; (2) redox signaling mechanisms by measuring reactive oxygen species (ROS) production, manganese superoxide dismutase (MnSOD) activity and mitochondrial membrane potential (MMP); (3) OPA1 and Mfn-1 expression that mediates the mitochondrial dynamics by PCR; and (4) mitochondrial morphology by MitoTracker Green FM staining. All three TiO2 nanoparticles induced a significant loss (p < 0.05) in hepatocyte functions even at concentrations as low as 50 ppm with commercially used P25 causing maximum damage. TiO2 nanoparticles induced a strong oxidative stress in primary hepatocytes. TiO2 nanoparticles exposure also resulted in morphological changes in mitochondria and substantial loss in the fusion process, thus impairing the mitochondrial dynamics. Although this study demonstrated that TiO2 nanoparticles exposure resulted in substantial damage to primary hepatocytes, more in vitro and in vivo studies are required to determine the complete toxicological mechanism in primary hepatocytes and subsequently liver function.


Psychopharmacological investigations of a lead-induced long-term cognitive deficit in monkeys.
This study investigated pharmacological manipulations of the cholinergic (ACh) and dopaminergic (DA) transmitter systems in monkeys with a long-term lead-induced cognitive deficit on delayed spatial alternation (DSA). Both ACh and DA have been found to be affected by developmental lead exposure and to be involved with performance on spatial learning and memory tasks. The lead-induced deficit in performance accuracy on DSA persisted throughout the 2 years of this experiment, which ended more than 8 years after the end of the postnatal lead exposure. Acute administration of agonists and antagonists of the ACh and DA systems did not elicit differential effects from the lead-exposed and control groups in terms of DSA per cent correct performance. The ACh antagonist, scopolamine, caused a dose-related decline in performance in both groups. Significant amelioration of the lead-induced DSA deficit was achieved by chronic treatment with the DA agonist, L-dopa. After withdrawal from L-dopa, the lead-related deficit reappeared. Improvement in performance of the lead-treated group was also seen after chronic amphetamine administration, but this effect was not significant. These data implicate DA mechanisms in the long-lasting cognitive effects of developmental lead exposure. The alleviation of the deficit with chronic administration of a DA precursor points to a possible line of treatment for the cognitive effects of developmental lead exposure.


Effects of response priming and inhibition on movement planning and execution.
The authors used a precueing method to examine the effects of response priming and inhibition on goal-directed action. Participants (N = 18) completed aiming movements to 1 of 2 locations following predictive (80% cued), nonpredictive (50% cued), and antipredictive (20% cued) precues at 1 of the 2 possible target locations. Consistent with previous research, participants responded more quickly to targets at cued locations than to targets at uncued locations in the 80% condition, and more quickly to targets presented at the uncued than to those presented at cued locations in the 50% and 20% conditions. As predicted by models of action-centered selective attention, movement trajectories deviated away from the cued location in the 50% condition. Movement trajectories were also altered in the 80% and the 20% conditions. Movements directed to the uncued location deviated away from the cued location in the 80% condition, whereas movements directed to the cued location deviated away from the uncued location in the 20% condition. The authors explain the latter trajectory results as a strategy of overcompensation.


Pragmatic model of patient satisfaction in general practice: progress towards a theory.
A major problem in the measurement of patient satisfaction is the lack of an adequate theory to explain the meaning of satisfaction, and hence how it should be measured and how the findings are interpreted. Because of the lack of a fully developed theory, when developing patient satisfaction questionnaires for use in general practice, a simple model was used. This model was pragmatic in that it linked together empirical evidence about patient satisfaction without recourse to more general social or psychological theory of behaviour, other than to define satisfaction as an attitude. Several studies with the questionnaires confirm in general the components of the model. However, the importance of personal care had not been sufficiently emphasised, and therefore the model has been revised. It can now serve as a basis for future research into patient satisfaction, in particular as a stimulus for investigating the links between components of the model and underlying psychological or other behavioural theories.


A rapamycin-sensitive pathway down-regulates insulin signaling via phosphorylation and proteasomal degradation of insulin receptor substrate-1.
Insulin receptor substrate-1 (IRS-1) is a major substrate of the insulin receptor and acts as a docking protein for Src homology 2 domain containing signaling molecules that mediate many of the pleiotropic actions of insulin. Insulin stimulation elicits serine/threonine phosphorylation of IRS-1, which produces a mobility shift on SDS-PAGE, followed by degradation of IRS-1 after prolonged stimulation. We investigated the molecular mechanisms and the functional consequences of these phenomena in 3T3-L1 adipocytes. PI 3-kinase inhibitors or rapamycin, but not the MEK inhibitor, blocked both the insulin-induced electrophoretic mobility shift and degradation of IRS-1. Adenovirus-mediated expression of a membrane-targeted form of the p110 subunit of phosphatidylinositol (PI) 3-kinase (p110CAAX) induced a mobility shift and degradation of IRS-1, both of which were inhibited by rapamycin. Lactacystin, a specific proteasome inhibitor, inhibited insulin-induced degradation of IRS-1 without any effect on its electrophoretic mobility. Inhibition of the mobility shift did not significantly affect tyrosine phosphorylation of IRS-1 or downstream insulin signaling. In contrast, blockade of IRS-1 degradation resulted in sustained activation of Akt, p70 S6 kinase, and mitogen-activated protein (MAP) kinase during prolonged insulin treatment. These results indicate that insulin-induced serine/threonine phosphorylation and degradation of IRS-1 are mediated by a rapamycin-sensitive pathway, which is downstream of PI 3-kinase and independent of ras/MAP kinase. The pathway leads to degradation of IRS-1 by the proteasome, which plays a major role in down-regulation of certain insulin actions during prolonged stimulation.


TECPR2 Associated Neuroaxonal Dystrophy in Spanish Water Dogs.
Clinical, pathological and genetic examination revealed an as yet uncharacterized juvenile-onset neuroaxonal dystrophy (NAD) in Spanish water dogs. Affected dogs presented with various neurological deficits including gait abnormalities and behavioral deficits. Histopathology demonstrated spheroid formation accentuated in the grey matter of the cerebral hemispheres, the cerebellum, the brain stem and in the sensory pathways of the spinal cord. Iron accumulation was absent. Ultrastructurally spheroids contained predominantly closely packed vesicles with a double-layered membrane, which were characterized as autophagosomes using immunohistochemistry. The family history of the four affected dogs suggested an autosomal recessive inheritance. SNP genotyping showed a single genomic region of extended homozygosity of 4.5 Mb in the four cases on CFA 8. Linkage analysis revealed a maximal parametric LOD score of 2.5 at this region. By whole genome re-sequencing of one affected dog, a perfectly associated, single, non-synonymous coding variant in the canine tectonin beta-propeller repeat-containing protein 2 (TECPR2) gene affecting a highly conserved region was detected (c.4009C>T or p.R1337W). This canine NAD form displays etiologic parallels to an inherited TECPR2 associated type of human hereditary spastic paraparesis (HSP). In contrast to the canine NAD, the spinal cord lesions in most types of human HSP involve the sensory and the motor pathways. Furthermore, the canine NAD form reveals similarities to cases of human NAD defined by widespread spheroid formation without iron accumulation in the basal ganglia. Thus TECPR2 should also be considered as candidate gene for human NAD. Immunohistochemistry and the ultrastructural findings further support the assumption, that TECPR2 regulates autophagosome accumulation in the autophagic pathways. Consequently, this report provides the first genetic characterization of juvenile canine NAD, describes the histopathological features associated with the TECPR2 mutation and provides evidence to emphasize the association between failure of autophagy and neurodegeneration.


Deep Dyspareunia: Review of Pathophysiology and Proposed Future Research Priorities.
Dyspareunia has been traditionally divided into superficial (introital) dyspareunia and deep dyspareunia (pain with deep penetration). While deep dyspareunia can coexist with a variety of conditions, recent work in endometriosis has demonstrated that coexistence does not necessarily imply causation. Therefore, a reconsideration of the literature is required to clarify the pathophysiology of deep dyspareunia. To review the pathophysiology of deep dyspareunia, and to propose future research priorities. A narrative review after appraisal of published frameworks and literature search with the terms (dyspareunia AND endometriosis), (dyspareunia AND deep), (dyspareunia AND (pathophysiology OR etiology)). Deep dyspareunia (present/absent or along a pain severity scale). The narrative review demonstrates potential etiologies for deep dyspareunia, including gynecologic-, urologic-, gastrointestinal-, nervous system-, psychological-, and musculoskeletal system-related disorders. These etiologies can be classified according to anatomic mechanism, such as contact with a tender pouch of Douglas, uterus-cervix, bladder, or pelvic floor, with deep penetration. Etiologies of deep dyspareunia can also be stratified into 4 categories, as previously proposed for endometriosis specifically, to personalize management: type I (primarily gynecologic), type II (nongynecologic comorbid conditions), type III (central sensitization and genito-pelvic pain/penetration disorder), and type IV (mixed). We also identified gaps in the literature, such as lack of a validated patient-reported questionnaire or an objective measurement tool for deep dyspareunia and clinical trials not powered for sexual outcomes. We propose the following research priorities for deep dyspareunia: deep dyspareunia measurement tools, inclusion of the population avoiding intercourse due to deep dyspareunia, nongynecologic conditions in the generation of deep dyspareunia, exploration of sociocultural factors, clinical trials with adequate power for deep dyspareunia outcomes, partner variables, female sexual response, pathways between psychological factors and deep dyspareunia, and personalized approaches to deep dyspareunia. Orr N, Wahl K, Joannou A, et al. Deep Dyspareunia: Review of Pathophysiology and Proposed Future Research Priorities. Sex Med Rev 2020;8:3-17.


Tomato-based food products for prostate cancer prevention: what have we learned?
Evidence derived from a vast array of laboratory studies and epidemiological investigations have implicated diets rich in fruits and vegetables with a reduced risk of certain cancers. However, these approaches cannot demonstrate causal relationships and there is a paucity of randomized, controlled trials due to the difficulties involved with executing studies of food and behavioral change. Rather than pursuing the definitive intervention trials that are necessary, the thrust of research in recent decades has been driven by a reductionist approach focusing upon the identification of bioactive components in fruits and vegetables with the subsequent development of single agents using a pharmacologic approach. At this point in time, there are no chemopreventive strategies that are standard of care in medical practice that have resulted from this approach. This review describes an alternative approach focusing upon development of tomato-based food products for human clinical trials targeting cancer prevention and as an adjunct to therapy. Tomatoes are a source of bioactive phytochemicals and are widely consumed. The phytochemical pattern of tomato products can be manipulated to optimize anticancer activity through genetics, horticultural techniques, and food processing. The opportunity to develop a highly consistent tomato-based food product rich in anticancer phytochemicals for clinical trials targeting specific cancers, particularly the prostate, necessitates the interactive transdisciplinary research efforts of horticulturalists, food technologists, cancer biologists, and clinical translational investigators.


Naked dense bodies provoke depression.
At presynaptic active zones (AZs), the frequently observed tethering of synaptic vesicles to an electron-dense cytomatrix represents a process of largely unknown functional significance. Here, we identified a hypomorphic allele, brpnude, lacking merely the last 1% of the C-terminal amino acids (17 of 1740) of the active zone protein Bruchpilot. In brpnude, electron-dense bodies were properly shaped, though entirely bare of synaptic vesicles. While basal glutamate release was unchanged, paired-pulse and sustained stimulation provoked depression. Furthermore, rapid recovery following sustained release was slowed. Our results causally link, with intramolecular precision, the tethering of vesicles at the AZ cytomatrix to synaptic depression.


Synergistic antitumor effect between gefitinib and fractionated irradiation in anaplastic oligodendrogliomas cannot be predicted by the Egfr signaling activity.
In high-grade gliomas, the identification of patients that could benefit from EGFR inhibitors remains a challenge, hindering the use of these agents. Using xenografts models, we evaluated the antitumor effect of the combined treatment "gefitinib + radiotherapy" and aimed to identify the profile of responsive tumors. Expression of phosphorylated proteins involved in the EGFR-dependent signaling pathways was analyzed in 10 glioma models. We focused on three models of anaplastic oligodendrogliomas (TCG2, TCG3 and TCG4) harboring high levels of phospho-EGFR, phospho-AKT and phospho-MEK1. They were treated with gefitinib (GEF 75 mg/kg/day x 5 days/week, for 2 weeks) and/or fractionated radiotherapy (RT: 5x2Gy/week for 2 weeks). Our results showed that GEF and/or RT induced significant tumor growth delays. However, only the TCG3 xenografts were highly responsive to the combination GEF+RT, with ∼50% of tumor cure. Phosphoproteins analysis five days after treatment onset demonstrated in TCG3 xenografts, but not in TCG2 model, that the EGFR-dependent pathways were inhibited after GEF treatment. Moreover, TCG3-bearing mice receiving GEF monotherapy exhibited a transient beneficial therapeutic response, rapidly followed by tumor regrowth, along with a major vascular remodeling. Taken together, our data evoked an "EGFR-addictive" behavior for TCG3 tumors. This study confirms that combination of gefitinib with fractionated irradiation could be a potent therapeutic strategy for anaplastic oligodendrogliomas harboring EGFR abnormalities but this treatment seems mainly beneficial for "EGFR-addictive" tumors. Unfortunately, neither the usual molecular markers (EGFR amplification, PTEN loss) nor the basal overexpression of phosphoproteins were useful to distinguish this responsive tumor. Evaluating the impact of TKIs on the EGFR-dependent pathways during the treatment might be more relevant, and requires further validation.


Neurotensin Is Coexpressed, Coreleased, and Acts Together With GLP-1 and PYY in Enteroendocrine Control of Metabolism.
The 2 gut hormones glucagon-like peptide-1 (GLP-1) and peptide YY (PYY) are well known to be coexpressed, costored, and released together to coact in the control of key metabolic target organs. However, recently, it became clear that several other gut hormones can be coexpressed in the intestinal-specific lineage of enteroendocrine cells. Here, we focus on the anatomical and functional consequences of the coexpression of neurotensin with GLP-1 and PYY in the distal small intestine. Fluorescence-activated cell sorting analysis, laser capture, and triple staining demonstrated that GLP-1 cells in the crypts become increasingly multihormonal, ie, coexpressing PYY and neurotensin as they move up the villus. Proglucagon promoter and pertussis toxin receptor-driven cell ablation and reappearance studies indicated that although all the cells die, the GLP-1 cells reappear more quickly than PYY- and neurotensin-positive cells. High-resolution confocal fluorescence microscopy demonstrated that neurotensin is stored in secretory granules distinct from GLP-1 and PYY storing granules. Nevertheless, the 3 peptides were cosecreted from both perfused small intestines and colonic crypt cultures in response to a series of metabolite, neuropeptide, and hormonal stimuli. Importantly, neurotensin acts synergistically, ie, more than additively together with GLP-1 and PYY to decrease palatable food intake and inhibit gastric emptying, but affects glucose homeostasis in a more complex manner. Thus, neurotensin is a major gut hormone deeply integrated with GLP-1 and PYY, which should be taken into account when exploiting the enteroendocrine regulation of metabolism pharmacologically.


Epithelioid Granulomas Associate With Increased Severity and Progression of Crohn's Disease, Based on 6-Year Follow-Up.
Epithelioid granulomas are characteristics of a subset of patients with Crohn's disease (CD), but their significance, with regard to disease progression and severity, is unclear. We investigated the relationship between granulomas and CD severity over a 6-year time period in a large cohort of patients. We performed a retrospective study of patients with CD seen at the Inflammatory Bowel Disease Center at the University of Pittsburgh; data were collected from 2009 through 2014 and patients were assigned to groups with and without histologic evidence of granuloma. Demographic, clinical (including disease activity, quality of life, medication use, and healthcare utilization), and laboratory data were used in association and survival analyses. Differences between groups were evaluated using the Mann-Whitney U-test for continuous variables. Of 1466 patients with CD, granulomas were identified in 187 (12.8%). In the subset of patients who underwent surgery, 21.0% had granulomas. The presence of granuloma was associated with increased serum levels of c-reactive protein (odds ratio [OR], 2.9; 95% CI, 2.078-4.208; P < .0001), younger mean age at diagnosis (23.6 ± 11.3 years in patients with granulomas vs 27.9 ± 13.3 years in patients without; P = .0005), higher rates of stricturing or penetrating disease phenotype, higher rates of steroid and narcotic use, and higher healthcare utilization. Among patients that underwent surgery, the presence of granulomas was associated with need for repeat surgery during the 6-year observation period (OR, 2.5; 95% CI, 1.54-4.02; P = .0002). Infliximab use was associated with detection of granuloma in a significantly lower proportion of surgical specimens compared to patients who had not been treated with a biologic agent (OR, 0.22; 95 CI, 0.05-0.97; P = .03). Epithelioid granulomas develop in less than 13% of patients with CD, and are associated with a more aggressive disease phenotype. Patients who have undergone surgery for CD and have granulomas are at increased risk for repeat surgery within 6 years.


Predictive and prognostic role of activated mammalian target of rapamycin in cervical cancer treated with cisplatin-based neoadjuvant chemotherapy.
The present study was designed to clarify the expression and prognostic significance of activated Akt and mTOR in cervical cancer and their correlation with response to neoadjuvant chemotherapy (NAC). Immunohistochemical analysis for p-Akt and p-mTOR expression was performed on paraffin-embedded biopsy specimens from 25 patients with advanced cervical cancer (stage Ib2-IIb). We correlated this finding with various clinicopathological variables and prognosis by uni- and multivariate analyses. All patients received cisplatin-based NAC, and primary tumor response was evaluated by RECIST criteria and then classified as a positive or negative response. Activation of Akt was detected in the cytoplasm and nucleus of the cancer cells in 12 patients (48%), whereas p-mTOR was detected in the cytoplasm and membrane of the cancer cells in 13 patients (52%). Post NAC evaluation of the primary tumor revealed 68% (17/25) responsive tumors. The expression of p-mTOR and distant metastasis significantly correlated with the response to NAC (p = 0.0101 and p = 0.0107); however, there was no significant correlation between p-Akt and p-mTOR expression and any of the clinicopathological characteristics of the patients. In the univariate analysis, activated Akt and mTOR were found to be significant prognostic indicators (p < 0.05). In multivariate analysis, p-mTOR expression retained its significance as an independent poor prognostic marker (p = 0.0178). In summary, our present study showed that cervical cancer expressed Akt and mTOR activation. Moreover, the expression of phosphorylated mTOR may have a role as a marker to predict response to chemotherapy and survival of cervical cancer patients who are treated with cisplatin-based neoadjuvant chemotherapy. Our results suggest that the mTOR cascade may be a promising target for therapeutic intervention in cervical cancer.


Derivation of algorithms for phase-shifting interferometry using the concept of a data-sampling window.
I propose a systematic way to derive efficient, error-compensating algorithms for phase-shifting interferometry by integer approximation of well-known data-sampling windows. The theoretical basi of the approach is the observation that many of the common sources of phase-estimation error can be related to the frequency-domain characteristics of the sampling window. Improving these characteristics can therefore improve the overall performance of the algorithm. Analysis of a seven-frame example algorithm demonstrates an exceptionally good resistance to first- and second-order distortions in the phase shift and a much reduced sensitivity to low-frequency mechanical vibration.


Inter-rater reliability between nurses for a new paediatric triage system based primarily on vital parameters: the Paediatric Triage Instrument (PETI).
The major paediatric triage systems are primarily based on flow charts involving signs and symptoms for orientation and subjective estimates of the patient's condition. In contrast, the 4-level Paediatric Triage Instrument (PETI) is primarily based on vital parameters and was developed exclusively for paediatric triage in patients with medical symptoms. The aim of this study was to assess the inter-rater reliability of this triage system in children when used by nurses. A design was employed in which triage was performed simultaneously and independently by a research nurse and an emergency department (ED) nurse using the PETI. All patients aged ≤12 years who presented at the ED with a medical symptom were considered eligible for participation. The 89 participants exhibited a median age of 2 years and were triaged by 28 different nurses. The inter-rater reliability between nurses calculated with the quadratic-weighted κ was 0.78 (95% CI 0.67 to 0.89); the linear-weighted κ was 0.67 (95% CI 0.56 to 0.80) and the unweighted κ was 0.59 (95% CI 0.44 to 0.73). For the patients aged <1, 1-3 and >3 years, the quadratic-weighted κ values were 0.67 (95% CI 0.39 to 0.94), 0.86 (95% CI 0.75 to 0.97) and 0.73 (95% CI 0.49 to 0.97), respectively. The median triage duration was 6 min. The PETI exhibited substantial reliability when used in children aged ≤12 years and almost perfect reliability among children aged 1-3 years. Moreover, rapid application of the PETI was demonstrated. This study has some limitations, including sample size and generalisability, but the PETI exhibited promise regarding reliability, and the next step could be either a larger reliability study or a validation study.


Large-aperture, equal-path interferometer for precision measurements of flat transparent surfaces.
The measurement of flat optical components often presents difficulties because the presence of parallel surfaces generates multiple reflections that confuse conventional laser-based interferometers. These same parts have increasingly demanding surface finish tolerances as technologies improve over time, further complicating the metrology task. Here we describe an interferometric optical system for high-accuracy noncontact evaluation of the form and texture of precision flat surfaces based on an equal-optical-path geometry that uses extended, broadband illumination to reduce the influence of speckle noise, multiple reflections, and coherent artifacts by a factor of 10 when compared to laser-based systems. Combined with a low-distortion, fixed-focus imaging system and 4-Mpixel camera, the 100 mm aperture instrument offers surface height resolutions of 0.1 nm over lateral spatial frequencies extending from 0.01 to 10 cycles/mm. The instrument is vibration resistant for production-line testing of flat optics such as glass hard disks for the data-storage industry and flat-panel-display substrates.


Online Simulation Training of Child Sexual Abuse Interviews With Feedback Improves Interview Quality in Japanese University Students.
Although previous research has confirmed the effectiveness of simulated child sexual abuse interviews with feedback, its validation is limited to Western contexts and face-to-face settings. The present study aims to extend this research to non-Western and online/remote training conditions. Thirty-two Japanese undergraduate students were randomly assigned to a control or feedback group. The feedback group conducted a set of six online simulated child sexual abuse interviews while receiving feedback after each interview in an attempt to improve the quality of their questioning style. The feedback consisted of the outcome of the alleged cases and the quality of the questions asked in the interviews. The control group conducted the interviews without feedback. The feedback (vs. control) increased the proportion of recommended questions (first interview: 45%; last interview: 65% vs. first: 43%; last: 42%, respectively) by using fewer not-recommended questions and eliciting fewer incorrect details. Furthermore, only participants in the feedback group (7 out of 17) demonstrated a reliable change in the proportion of recommended questions. The present study explores the efficacy of simulated interview training with avatars in a different cultural setting and in the context of remote administration. The differences between the present study and previous research are discussed in light of cultural and logistical aspects.


Racial/Ethnic Disparities Among Extremely Preterm Infants in the United States From 2002 to 2016.
Racial/ethnic disparities in quality of care among extremely preterm infants are associated with adverse outcomes. To assess whether racial/ethnic disparities in major outcomes and key care practices were changing over time among extremely preterm infants. This observational cohort study used prospectively collected data from 25 US academic medical centers. Participants included 20 092 infants of 22 to 27 weeks' gestation with a birth weight of 401 to 1500 g born at centers participating in the National Institute of Child Health and Human Development Neonatal Research Network from 2002 to 2016. Of these infants, 9316 born from 2006 to 2014 were eligible for follow-up at 18 to 26 months' postmenstrual age (excluding 5871 infants born before 2006, 2594 infants born after 2014, and 2311 ineligible infants including 64 with birth weight >1000 g and 2247 infants with gestational age >26 6/7 weeks), of whom 745 (8.0%) did not have known follow-up outcomes at 18 to 26 months. Rates of mortality, major morbidities, and care practice use over time were evaluated using models adjusted for baseline characteristics, center, and birth year. Data analyses were conducted from 2018 to 2019. In total, 20 092 infants with a mean (SD) gestational age of 25.1 (1.5) weeks met the inclusion criteria and were available for the primary outcome: 8331 (41.5%) black infants, 3701 (18.4%) Hispanic infants, and 8060 (40.1%) white infants. Hospital mortality decreased over time in all groups. The rate of improvement in hospital mortality over time did not differ among black and Hispanic infants compared with white infants (black infants went from 35% to 24%, Hispanic infants went from 32% to 27%, and white infants went from 30% to 22%; P = .59 for race × year interaction). The rates of late-onset sepsis among black infants (went from 37% to 24%) and Hispanic infants (went from 45% to 23%) were initially higher than for white infants (went from 36% to 25%) but decreased more rapidly and converged during the most recent years (P = .02 for race × year interaction). Changes in rates of other major morbidities did not differ by race/ethnicity. Death before follow-up decreased over time (from 2006 to 2014: black infants, 14%; Hispanic infants, 39%, white infants, 15%), but moderate-severe neurodevelopmental impairment increased over time in all racial/ethnic groups (increase from 2006 to 2014: black infants, 70%; Hispanic infants, 123%; white infants, 130%). Rates of antenatal corticosteroid exposure (black infants went from 72% to 90%, Hispanic infants went from 73% to 83%, and white infants went from 86% to 90%; P = .01 for race × year interaction) and of cesarean delivery (black infants went from 45% to 59%, Hispanic infants went from 49% to 59%, and white infants went from 62% to 63%; P = .03 for race × year interaction) were initially lower among black and Hispanic infants compared with white infants, but these differences decreased over time. Among extremely preterm infants, improvements in adjusted rates of mortality and most major morbidities did not differ by race/ethnicity, but rates of neurodevelopmental impairment increased in all groups. There were narrowing racial/ethnic disparities in important care practices, including the use of antenatal corticosteroids and cesarean delivery.


Land plants equilibrate O2 and CO2 concentrations in the atmosphere.
The role of land plants in establishing our present day atmosphere is analysed. Before the evolution of land plants, photosynthesis by marine and fresh water organisms was not intensive enough to deplete CO(2) from the atmosphere, the concentration of which was more than the order of magnitude higher than present. With the appearance of land plants, the exudation of organic acids by roots, following respiratory and photorespiratory metabolism, led to phosphate weathering from rocks thus increasing aquatic productivity. Weathering also replaced silicates by carbonates, thus decreasing the atmospheric CO(2) concentration. As a result of both intensive photosynthesis and weathering, CO(2 )was depleted from the atmosphere down to low values approaching the compensation point of land plants. During the same time period, the atmospheric O(2) concentration increased to maximum levels about 300 million years ago (Permo-Carboniferous boundary), establishing an O(2)/CO(2) ratio above 1000. At this point, land plant productivity and weathering strongly decreased, exerting negative feedback on aquatic productivity. Increased CO(2) concentrations were triggered by asteroid impacts and volcanic activity and in the Mesozoic era could be related to the gymnosperm flora with lower metabolic and weathering rates. A high O(2)/CO(2) ratio is metabolically linked to the formation of citrate and oxalate, the main factors causing weathering, and to the production of reactive oxygen species, which triggered mutations and stimulated the evolution of land plants. The development of angiosperms resulted in a decrease in CO(2) concentration during the Cenozoic era, which finally led to the glacial-interglacial oscillations in the Pleistocene epoch. Photorespiration, the rate of which is directly related to the O(2)/CO(2) ratio, due to the dual function of Rubisco, may be an important mechanism in maintaining the limits of O(2) and CO(2) concentrations by restricting land plant productivity and weathering.


The six minute walk test accurately estimates mean peak oxygen uptake.
Both Peak Oxygen Uptake (peak VO2), from cardiopulmonary exercise testing (CPET) and the distance walked during a Six-Minute Walk Test (6 MWD) are used for following the natural history of various diseases, timing of procedures such as transplantation and for assessing the response to therapeutic interventions. However, their relationship has not been clearly defined. We determined the ability of 6 MWD to predict peak VO2 using data points from 1,083 patients with diverse cardiopulmonary disorders. The patient data came from a study we performed and 10 separate studies where we were able to electronically convert published scattergrams to bivariate points. Using Linear Mixed Model analysis (LMM), we determined what effect factors such as disease entity and different inter-site testing protocols contributed to the magnitude of the standard error of estimate (SEE). The LMM analysis found that only 0.16 ml/kg/min or about 4% of the SEE was due to all of the inter-site testing differences. The major source of error is the inherent variability related to the two tests. Therefore, we were able to create a generalized equation that can be used to predict peak VO2 among patients with different diseases, who have undergone various exercise protocols, with minimal loss of accuracy. Although 6 MWD and peak VO2 are significantly correlated, the SEE is unacceptably large for clinical usefulness in an individual patient. For the data as a whole it is 3.82 ml/kg/min or 26.7% of mean peak VO2. Conversely, the SEE for predicting the mean peak VO2 from mean 6 MWD for the 11 study groups is only 1.1 ml/kg/min. A generalized equation can be used to predict peak VO2 from 6 MWD. Unfortunately, like other prediction equations, it is of limited usefulness for individual patients. However, the generalized equation can be used to accurately estimate mean peak VO2 from mean 6 MWD, among groups of patients with diverse diseases without the need for cardiopulmonary exercise testing. The equation is:


Influence of dietary linoleic acid on leucocyte sodium transport and blood pressure.
In a randomised double blind study to determine whether an increase in the polyunsaturated fat linoleic acid might influence leucocyte membrane sodium transport 22 normotensive volunteers received an oral supplement of linoleic acid or placebo daily for four weeks. Mean total sodium efflux rose significantly during supplementation with linoleic acid compared with placebo. In addition, all components of lying and standing blood pressure fell, though only the fall in supine systolic pressure was significant. Dietary supplementation with linoleic acid may alter ion fluxes across the cell membrane, presumably through changes in its physicochemical structure. In addition, the change in fat intake may lower blood pressure, though to only a very modest extent.


Insights into gastric neuroendocrine tumors burden.
Type 1 gastric neuroendocrine tumors (gNETs) are usually small lesions, restricted to mucosal and sub-mucosal layers of corpus and fundus, with low aggressive behavior, for the majority of cases. Nevertheless, some cases present aggressive behavior. The increasing incidence of gNETs brings together a new relevant problem: how to identify potentially aggressive type 1 gNETs. The challenging problem seems to be finding out signs or features able to predict potentially aggressive cases, allowing a tailored approach, since the involved societies dedicated to provide guidelines for management of these neoplasms apparently failed in producing staging systems able to accurately predict prognosis of these tumors. Additionally, it is also important to try to find out explanations for increasing incidence, as well as to identify potential targets aiming to reach better control of this neoplasia. Here, we discuss potential pathways implicated in aggressive behavior, as well as new strategies to improve clinical management of these tumors.


The ecological genetics of introduced populations of the giant toad Bufo marinus. II. Effective population size.
The allele frequencies are described at ten polymorphic enzyme loci (of a total of 22 loci sampled) in 15 populations of the neotropical giant toad, Bufo marinus, introduced to Hawaii and Australia in the 1930s. The history of establishment of the ten populations is described and used as a framework for the analysis of allele frequency variances. The variances are used to determine the effective sizes of the populations. The estimates obtained (390 and 346) are reasonably precise, homogeneous between localities and much smaller than estimates of neighborhood size obtained previously using ecological methods. This discrepancy is discussed, and it is concluded that the estimates obtained here using genetic methods are the more reliable.


The health equity dimensions of urban food systems.
There is increasing recognition that the nutrition transition sweeping the world's cities is multifaceted. Urban food and nutrition systems are beginning to share similar features, including an increase in dietary diversity, a convergence toward "Western-style" diets rich in fat and refined carbohydrate and within-country bifurcation of food supplies and dietary conventions. Unequal access to the available dietary diversity, calories, and gastronomically satisfying eating experience leads to nutritional inequalities and diet-related health inequities in rich and poor cities alike. Understanding the determinants of inequalities in food security and nutritional quality is a precondition for developing preventive policy responses. Finding common solutions to under- and overnutrition is required, the first step of which is poverty eradication through creating livelihood strategies. In many cities, thousands of positions of paid employment could be created through the establishment of sustainable and self-sufficient local food systems, including urban agriculture and food processing initiatives, food distribution centers, healthy food market services, and urban planning that provides for multiple modes of transport to food outlets. Greater engagement with the food supply may dispel many of the food anxieties affluent consumers are experiencing.


Meniscus formation in a capillary and the role of contact line friction.
We studied spontaneous formation of an internal meniscus by dipping glass capillaries of 25 μm to 350 μm radii into low volatile hexadecane and tributyl phosphate. X-ray phase contrast and high speed optical microscopy imaging were employed. We showed that the meniscus completes its formation when the liquid column is still shorter than the capillary radius. After that, the meniscus travels about ten capillary radii at a constant velocity. We demonstrated that the experimental observations can be explained by introducing a friction force linearly proportional to the meniscus velocity with a friction coefficient depending on the air/liquid/solid triplet. It was demonstrated that the friction coefficient does not depend on the capillary radius. Numerical solution of the force balance equation revealed four different uptake regimes that can be specified in a phase portrait. This phase portrait was found to be in good agreement with the experimental results and can be used as a guide for the design of thin porous absorbers.


Rates of DSM-IV-TR Trauma Exposure and Posttraumatic Stress Disorder Among Newly Matriculated College Students.
The negotiation of the freedoms and responsibilities introduced as adolescents begin college may be particularly challenging for those with a trauma history and traumatic stress sequelae (posttraumatic stress disorder; PTSD). The present study examined the prevalence of and risk for trauma and PTSD in a large sample of college students. Matriculating students (N = 3,014; 1,763 female, 1,251 male) at two U.S. universities completed online and paper assessments. Sixty-six percent reported exposure to a Criterion A trauma. Nine percent met criteria for PTSD. Female gender was a risk factor for trauma exposure. Gender and socioeconomic status (SES) were associated with trauma severity. Although in bivariate models, gender and SES were associated with PTSD, multivariate analyses suggested this risk was a function of trauma severity. Thus, students enter college with significant trauma histories and PTSD symptoms. Findings highlight the potential for outreach to incoming students with trauma and point to research directions to enhance understanding of the psychological needs of entering college students.


Pretreatment with low-dose fimasartan ameliorates NLRP3 inflammasome-mediated neuroinflammation and brain injury after intracerebral hemorrhage.
Nucleotide-binding and oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome, which is composed of an NLRP3 domain, the adaptor molecule apoptosis-associated speck-like protein containing a CARD (ASC) domain, and procaspase-1, plays an important role in the immune pathophysiology of the secondary damage induced by intracerebral hemorrhage (ICH). This study aims to investigate whether pre-stroke treatment with fimasartan, an angiotensin II receptor blocker, has anti-inflammatory effects on ICH by inhibiting the activation of the NLRP3 inflammasome. Sprague-Dawley rats were divided into five groups: sham, vehicle, low-dose (0.5 mg/kg) and regular-doses (1.0 and 3.0 mg/kg) fimasartan. These rats were treated for 30 days before the induction of collagenase-induced ICH and continuously 3 days after surgery. The mean blood pressure (BP) in the low-dose fimasartan group was not significantly different from that of control, and BP in the regular-dose groups was decreased in a dose-dependent manner. Pretreatment with low-dose fimasartan attenuated ICH-induced edema and improved neurological functions. Activation of the NLRP3/ASC/caspase-1 and the NF-κB pathways after ICH was markedly reduced by low-dose fimasartan. The double immunofluorescence staining of brain cells showed a significant decrease in the co-localization of NLRP3 with Iba1 (microglia marker) positive cells by fimasartan treatment. Cultured microglia cells stimulated by hemolysate demonstrated significant activation of the inflammasome, which was reduced by fimasartan. Pretreatment with a low-dose fimasartan alleviated brain damage after acute ICH by inhibiting the NLRP3 inflammasome without lowering MBP. Our study suggests pre-stroke administration of fimasartan could potentially attenuate ICH-induced secondary brain injury by targeting the inflammasome.


Sex hormones in hereditary angioneurotic oedema.
The fluctuations in sex hormone levels at the beginning of adolescence, in the perimenopausal period, during pregnancy or during the use of oral contraceptives can precipitate oedematous attacks in hereditary angioneurotic oedema (HANO). Attacks usually disappear after the onset of menopause. This study was undertaken to establish any relationship between the serum levels of sex hormones and the incidence of HANO attacks. Serum levels of LH, FSH, progesterone, oestradiol, testosterone, PRL and SHBG were measured in 78 patients [mean age 30.3 years (range 4-70 years)] with HANO. A questionnaire was used to explore the medical history of adult patients to characterize the evolution and the characteristics of attacks. The number of attacks was significantly higher [odds ratio (OR) 6.36 (1.31-30.81); P = 0.022] in females with high progesterone levels (> or = 4 nmol/l), irrespective of age, menstrual cycle and danazol dose. The OR was even higher [13.4 (2.2-81.4); P = 0.005] when only subcutaneous attacks were considered. Multiple logistic regression analysis demonstrated a significantly lower attack frequency during 1-year follow-up in patients with a higher (40 nmol/l) SHBG level (OR 0.25 (0.07-0.90); P = 0.034). This difference existed independently of age and danazol dose. In view of these results, the monitoring of progesterone and SHBG levels can prove useful in the prediction of attacks in hereditary angioneurotic oedema.


Daily and seasonal mitochondrial protection: Unraveling common possible mechanisms involving vitamin D and melatonin.
From an evolutionary point of view, vitamin D and melatonin appeared very early and share functions related to defense mechanisms. In the current clinical setting, vitamin D is exclusively associated with phosphocalcic metabolism. Meanwhile, melatonin has chronobiological effects and influences the sleep-wake cycle. Scientific evidence, however, has identified new actions of both molecules in different physiological and pathological settings. The biosynthetic pathways of vitamin D and melatonin are inversely related relative to sun exposure. A deficiency of these molecules has been associated with the pathogenesis of cardiovascular diseases, including arterial hypertension, neurodegenerative diseases, sleep disorders, kidney diseases, cancer, psychiatric disorders, bone diseases, metabolic syndrome, and diabetes, among others. During aging, the intake and cutaneous synthesis of vitamin D, as well as the endogenous synthesis of melatonin are remarkably depleted, therefore, producing a state characterized by an increase of oxidative stress, inflammation, and mitochondrial dysfunction. Both molecules are involved in the homeostatic functioning of the mitochondria. Given the presence of specific receptors in the organelle, the antagonism of the renin-angiotensin-aldosterone system (RAAS), the decrease of reactive species of oxygen (ROS), in conjunction with modifications in autophagy and apoptosis, anti-inflammatory properties inter alia, mitochondria emerge as the final common target for melatonin and vitamin D. The primary purpose of this review is to elucidate the common molecular mechanisms by which vitamin D and melatonin might share a synergistic effect in the protection of proper mitochondrial functioning.


Mutant fibrinogen cleared from the endoplasmic reticulum via endoplasmic reticulum-associated protein degradation and autophagy: an explanation for liver disease.
The endoplasmic reticulum (ER) quality control processes recognize and remove aberrant proteins from the secretory pathway. Several variants of the plasma protein fibrinogen are recognized as aberrant and degraded by ER-associated protein degradation (ERAD), thus leading to hypofibrinogenemia. A subset of patients with hypofibrinogenemia exhibit hepatic ER accumulation of the variant fibrinogens and develop liver cirrhosis. One such variant named Aguadilla has a substitution of Arg375 to Trp in the gamma-chain. To understand the cellular mechanisms behind clearance of the aberrant Aguadilla gamma-chain, we expressed the mutant gammaD domain in yeast and found that it was cleared from the ER via ERAD. In addition, we discovered that when ERAD was saturated, aggregated Aguadilla gammaD accumulated within the ER while a soluble form of the polypeptide transited the secretory pathway to the trans-Golgi network where it was targeted to the vacuole for degradation. Examination of Aguadilla gammaD in an autophagy-deficient yeast strain showed stabilization of the aggregated ER form, indicating that these aggregates are normally cleared from the ER via the autophagic pathway. These findings have clinical relevance in the understanding of and treatment for ER storage diseases.


Valproic acid-mediated neuroprotection and neurogenesis after spinal cord injury: from mechanism to clinical potential.
Spinal cord injury (SCI) is difficult to treat because of secondary injury. Valproic acid (VPA) is clinically approved for mood stabilization, but also counteracts secondary damage to functionally rescue SCI in animal models by improving neuroprotection and neurogenesis via inhibition of HDAC and GSK-3. However, a comprehensive review summarizing the therapeutic benefits and mechanisms of VPA for SCI and the issues affecting clinical trials is lacking, limiting future research on VPA and impeding its translation into clinical therapy for SCI. This article presents the current status of VPA treatment for SCI, emphasizing interactions between enhanced neuroprotection and neurogenesis. Crucial issues are discussed to optimize its clinical potential as a safe and effective treatment for SCI.


Shedding light on cell signaling: interpretation of FRET biosensors.
This Perspective compares recently developed approaches for studying live-cell signaling dynamics. It is now possible not only to study the changing localizations of proteins within living cells but to correlate these dynamics with quantitation of protein activities, including ligand interactions and phosphorylation. Initial applications of an increasingly varied toolchest of techniques have revealed strengths and weaknesses in each approach, clarifying which tool is best for which job.


Post-receptor mechanisms underlying striatal long-term depression.
Extracellular and intracellular recordings were obtained from striatal neurons in a brain slice preparation in order to characterize the post-receptor mechanisms underlying striatal posttetanic long-term depression (LTD). Striatal LTD was blocked in neurons intracellularly recorded either with 1,2-bis (o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA) or with EGTA, calcium (Ca2+) chelators. Intracellular injection of QX-314, a lidocaine derivative that has been shown to block voltage-dependent sodium channels, abolished action potential discharge and blocked striatal LTD. However, under this condition, striatal LTD was restored when, immediately before the delivery of the tetanus, the cell was depolarized at a membrane potential ranging between -30 mV and -20 mV by injecting continuous positive current. Nifedipine (10 microM), a blocker of voltage-dependent L-type Ca2+ channels, blocked striatal LTD. Nifedipine by itself altered neither cortically evoked EPSPs nor input resistance and firing properties of most of the recorded cells. Striatal LTD was also reduced or blocked by incubation of the slices in the presence of the following inhibitors of Ca(2+)-dependent protein kinases: staurosporine (10-50 nM), 1-(5-isoquinolinesulfonyl)-2- methylpiperazine (H-7; 10-50 microM), and calphostin C (1 microM). Our findings suggest that generation of striatal LTD requires a Ca2+ influx through voltage-dependent nifedipine-sensitive Ca2+ channels and a sufficient intracellular free Ca2+ concentration. Furthermore, this form of synaptic plasticity seems to involve the activation of Ca(2+)-dependent protein kinases. Different drugs, acting at receptor and/or post-receptor level, may affect this form of synaptic plasticity and might alter the formation of motor memory.


The Effects of Providing Fixed Compensation and Lottery-Based Rewards on Uptake of Medical Male Circumcision in Kenya: A Randomized Trial.
Effective demand creation strategies are needed to increase uptake of medical male circumcision and reduce new HIV infections in eastern and southern Africa. Building on insights from behavioral economics, we assessed whether providing compensation for opportunity costs of time or lottery-based rewards can increase male circumcision uptake in Kenya. Uncircumcised men aged 21-39 years were randomized in 1:1:1 ratio to 2 intervention groups or a control group. One intervention group was offered compensation of US $12.50 conditional on circumcision uptake. Compensation was provided in the form of food vouchers. A second intervention group was offered the opportunity to participate in a lottery with high-value prizes on undergoing circumcision. The primary outcome was circumcision uptake within 3 months. Among 903 participants enrolled, the group that received compensation of US $12.50 had the highest circumcision uptake (8.4%, 26/308), followed by the lottery-based rewards group (3.3%, 10/302), and the control group (1.3%, 4/299). Logistic regression analysis showed that compared with the control group, the fixed compensation group had significantly higher circumcision uptake [adjusted odds ratio 7.1; 95% CI: 2.4 to 20.8]. The lottery-based rewards group did not have significantly higher circumcision uptake than the control group (adjusted odds ratio 2.5; 95% CI: 0.8 to 8.1). Providing compensation was effective in increasing circumcision uptake among men over a short period. The results are consistent with studies showing that such interventions can modify health behaviors by addressing economic barriers and behavioral biases in decision making. Contrary to findings from studies of other health behaviors, lottery-based rewards did not significantly increase circumcision uptake. Registry for International Development Impact Evaluations: RIDIE-STUDY-ID-530e60df56107.