Cannabis use in people with Parkinson's disease and Multiple Sclerosis: A web-based investigation.
Cannabis has been used for medicinal purpose for thousands of years; however the positive and negative effects of cannabis use in Parkinson's disease (PD) and Multiple Sclerosis (MS) are mostly unknown. Our aim was to assess cannabis use in PD and MS and compare results of self-reported assessments of neurological disability between current cannabis users and non-users. An anonymous web-based survey was hosted on the Michael J. Fox Foundation and the National Multiple Sclerosis Society webpages from 15 February to 15 October 2016. The survey collected demographic and cannabis use information, and used standardized questionnaires to assess neurological function, fatigue, balance, and physical activity participation. Analysis of variance and chi-square tests were used for the analysis. The survey was viewed 801 times, and 595 participants were in the final data set. Seventy-six percent and 24% of the respondents reported PD and MS respectively. Current users reported high efficacy of cannabis, 6.4 (SD 1.8) on a scale from 0 to 7 and 59% reported reducing prescription medication since beginning cannabis use. Current cannabis users were younger and less likely to be classified as obese (P < 0.035). Cannabis users reported lower levels of disability, specifically in domains of mood, memory, and fatigue (P<0.040). Cannabis may have positive impacts on mood, memory, fatigue, and obesity status in people with PD and MS. Further studies using clinically and longitudinally assessed measurements of these domains are needed to establish if these associations are causal and determine the long-term benefits and consequences of cannabis use in people with PD and MS.


Assessment of commercial enzyme immunoassay for hepatitis C virus serotyping.
To assess a commercial enzyme immunoassay (EIA) for the serotyping of hepatitis C virus (HCV) for routine use in a diagnostic laboratory setting, as well as for noting the serotype prevalence of selected specimens. Seventy six serum specimens, submitted to the laboratory for routine hepatitis studies between May 1992 and February 1996 and stored at -20 degrees C, were evaluated. These specimens were categorised into specific hepatic, renal, and paediatric clinical conditions. The specimens all tested positive for HCV antibodies on a screening EIA, with confirmation on a recombinant immunoblot assay (RIBA). Certain specimens were also HCV RNA positive by the reverse transcription polymerase chain reaction (RT-PCR). All the specimens were serotyped using the newly developed serotyping EIA. Twenty seven (35.5%) specimens were typable. Type 5 predominated (56%), followed by type 1 (33%), types 1 and 6 (7%) and type 3 (4%). The serotype 5 specimens showed 85% and 90% reactivity with recombinant antigens c100-3 and c22-3c, respectively; serotype 1 specimens showed 75% and 100% reactivity with these antigens. All serotype 5 specimens reacted with the c33-c antigen, but only 60% of serotype 1 specimens reacted with this antigen. The differences in the reactivity of the serotype 5 and serotype 1 specimens for c33-c antigen in the RIBA were significant, but no significant differences in reactivity for antigens c-1-1, c100-3, and c22-3 were noted. Serotype 3 specimens showed equal reactivity with all four antigens used in the RIBA. The serotyping EIA was easy to use, rapid, and cost effective compared with molecular assays. This assay seems to be ideal for the routine diagnostic laboratory setting, but could not be used for certain clinical specimens. The demonstration of serotypes 5, 1, and 3 was not unexpected in this cohort. The occurrence of serotype 6, although concurrent and more likely to be a false cross reaction with serotype 1 peptides, requires confirmation by molecular genotyping before it can be claimed that this type is present in South Africa.


Potential use of spectral image analysis for the quantitative evaluation of estrogen receptors in breast cancer.
Evaluation of estrogen receptor (ER) content is an important factor in the choice of therapy and prognosis of breast cancer patients. In this study, we demonstrate a new spectral image analysis technique for objective and quantitative evaluations of stained specimens. The SpectraCube system was used to analyze nuclear antigens in thirteen cases of breast cancer stained by the immunoperoxidase method with hematoxylin counterstain. Spectral imaging segregated the spectrum of diaminobenzidine (DAB) from the background color of hematoxylin and a spectral index was calculated. The spectral index essentially agreed with the pathologist's index (on a scale of 0 to 3) in seven out of the thirteen cases. A substantial number of ER positive pixels was detected in the two cases scored as 0 by the pathologist's index. In a test case scored as 1 by the pathologist's index we detected a significant number of pixels, representing 47% of the nuclei, with DAB-intensity values higher than the cut-off value of 1.2. These data suggest that spectral image analysis is a sensitive method providing intensive information with high reproducibility. Our spectral imaging method is highly flexible, enabling the user to define the spatial resolution of the analyzed specimen by choosing the number of pixels per one nucleus.


Comparison of cell survival models for mixed LET radiation.
Biophysical models for predicting survival for mixed LET radiations have been investigated by comparisons with experimental results from heavy ion irradiations. The aim was to choose a model for further theoretical studies on the effects of a variable RBE for protons. Predicted survival curves by the Katz track-structure model, the linear quadratic model, LQ model, by Kellerer and Rossi and the lesion additivity model of Lam were compared to experimental survival curves for V79 cells that were irradiated with a mixture of nitrogen ions with an LET of either 78 or 165 keV/microm and 60Co gamma-rays. Results showed that all three models could predict survival within the uncertainty of the measurements for the different mixed radiation schedules used in this study. The choice of model could be made on other grounds, such as the type of model parameters and the availability of biological data for these parameters. Also, the possibility of including dose-rate effects and repair functions should be considered. For the purpose of carrying out theoretical studies on the effects of a variable RBE for protons, the LQ model was preferred.


Factors associated with multiple hospital readmissions for individuals with spinal cord injury.
To identify patient-level and clinical factors associated with multiple hospital readmissions in individuals with spinal cord injury (SCI). Retrospective cohort analysis of the publicly available dataset from the SCI Rehabilitation (SCIRehab) study. Six rehabilitation centers in the U.S. that participated in the SCIRehab study. Participants: Individuals with traumatic SCI (N=1371) who were consecutively enrolled in the SCIRehab study. The primary outcome was all-cause hospital readmission within 1 year of discharge from a rehabilitation center. The patient-level and clinical factors include employment status, depression, caregiver support, state-funded insurance, functional status, and rehabilitation services. Of the 1170 participants included in the study, 228 were readmitted once and 120 were readmitted multiple times. In our study, 34.2% and 10.8% were readmitted more than once due to genitourinary and respiratory conditions, respectively. Lower utilization of rehabilitation services, lower functional status, unemployment, and depression were associated with higher odds of being readmitted multiple times within one year of discharge from inpatient rehabilitation facility. Psychosocial and clinical factors were associated with increased risk for multiple readmissions in individuals with SCI. In order to reduce recurrent readmissions in individuals with SCI, further research is necessary to maximize efficacy of risk factor modification and prevention strategies.


Primary afferents evoke excitatory amino acid receptor-mediated EPSPs that are modulated by presynaptic GABAB receptors in lamprey.
1. The primary afferent neurons (dorsal cells) are of two types in lamprey, which are fast (touch) and slowly adapting (pressure), respectively. Intracellular stimulation of such sensory neurons evokes mono- and polysynaptic excitatory postsynaptic potentials (EPSPs) in spinobulbar neurons (giant interneurons) and in unidentified interneurons. Paired intracellular recordings between identified sensory cells and spinobulbar neurons made it possible to study the synaptic transmission in detail. It is shown that both touch and pressure primary afferents utilize excitatory amino acid (EAA) transmission and, furthermore, that these effects are subject to a presynaptic GABAB receptor modulation. 2. The monosynaptic mixed electrical and chemical EPSPs in giant interneurons had a mean peak amplitude of 3.2 +/- 1.3 (SD) mV, a time to peak of 4.7 +/- 1.2 ms, and a duration at one-half peak amplitude of 9.4 +/- 3.2 ms. Corresponding results were obtained with dorsal root or dorsal column stimulation. Seventy percent of the fast-adapting dorsal cells of the "touch" type evoked monosynaptic mixed EPSPs in giant interneurons, whereas only 3% of the slowly adapting "pressure" dorsal cells did. 3. The chemical part of the monosynaptic EPSPs evoked in giant interneurons was, in all cases tested, blocked by application of EAA antagonists, like the nonselective antagonist kynurenic acid (KYAC; 2 mM). The selective kainate/alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 5 microM) had a similar effect, whereas the selective N-methyl-D-aspartate (NMDA) receptor antagonist 2-aminophosphono-5-valeric acid (AP-5; 200-400 microM) did not change the EPSP, even in the absence of magnesium ions. 4. The monosynaptic excitatory synaptic transmission was modulated by application of the selective GABAB receptor agonist L-baclofen (5-10 mM local droplet application or 100-1,000 microM bath applied) or by gamma-aminobutyric acid (GABA; 100-1,000 microM), also when GABAA receptor-evoked effects were blocked by bicuculline (10 microM). L-baclofen or GABA in combination with bicuculline did not evoke any effects in the postsynaptic neuron on membrane potential, input resistance, or spike threshold. Therefore the effects of the GABAB receptor activation most likely occurs at the presynaptic afferent level. 5. In conclusion, the monosynaptic excitation from skin mechanoreceptors evoked in spinobulbar neurons is mediated by EAA receptors of the kainate/AMPA type. GABAB receptor activation causes a depression of this EPSP, most likely because of a presynaptic action. GABA interneurons are known to form close appositions on sensory axons in the lamprey.


Scaling of caterpillar body properties and its biomechanical implications for the use of a hydrostatic skeleton.
Caterpillars can increase their body mass 10,000-fold in 2 weeks. It is therefore remarkable that most caterpillars appear to maintain the same locomotion kinematics throughout their entire larval stage. This study examined how the body properties of a caterpillar might change to accommodate such dramatic changes in body load. Using Manduca sexta as a model system, we measured changes in body volume, tissue density and baseline body pressure, and the dimensions of load-bearing tissues (the cuticle and muscles) over a body mass range from milligrams to several grams. All Manduca biometrics relevant to the hydrostatic skeleton scaled allometrically but close to the isometric predictions. Body density and pressure were almost constant. We next investigated the effects of scaling on the bending stiffness of the caterpillar hydrostatic skeleton. The anisotropic non-linear mechanical response of Manduca muscles and soft cuticle has previously been quantified and modeled with constitutive equations. Using biometric data and these material laws, we constructed finite element models to simulate a hydrostatic skeleton under different conditions. The results show that increasing the internal pressure leads to a non-linear increase in bending stiffness. Increasing the body size results in a decrease in the normalized bending stiffness. Muscle activation can double this stiffness in the physiological pressure range, but thickening the cuticle or increasing the muscle area reduces the structural stiffness. These non-linear effects may dictate the effectiveness of a hydrostatic skeleton at different sizes. Given the shared anatomy and size variation in Lepidoptera larvae, these mechanical scaling constraints may implicate the diverse locomotion strategies in different species.


Is there a need for a specific educational scholarship for using e-learning in medical education?
We propose the need for a specific educational scholarship when using e-learning in medical education. Effective e-learning has additional factors that require specific critical attention, including the design and delivery of e-learning. An important aspect is the recognition that e-learning is a complex intervention, with several interconnecting components that have to be aligned. This alignment requires an essential iterative development process with usability testing. Effectiveness of e-learning in one context may not be fully realized in another context unless there is further consideration of applicability and scalability. We recommend a participatory approach for an educational scholarship for using e-learning in medical education, such as by action research or design-based research.


Treatment for Word Retrieval in Semantic and Logopenic Variants of Primary Progressive Aphasia: Immediate and Long-Term Outcomes.
Purpose Recent studies confirm the utility of speech-language intervention in primary progressive aphasia (PPA); however, long-term outcomes, ideal dosage parameters, and relative benefits of intervention across clinical variants warrant additional investigation. The purpose of this study was to determine whether naming treatment affords significant, lasting, and generalized improvement for individuals with semantic and logopenic PPA and whether dosage manipulations significantly affect treatment outcomes. Method Eighteen individuals with PPA (9 semantic and 9 logopenic variant) underwent lexical retrieval treatment designed to leverage spared cognitive-linguistic domains and develop self-cueing strategies to promote naming. One group (n = 10) underwent once-weekly treatment sessions, and the other group (n = 8) received the same treatment with 2 sessions per week and an additional "booster" treatment phase at 3 months post-treatment. Performance on trained and untrained targets/tasks was measured immediately after treatment and at 3, 6, and 12 months post-treatment. Results Outcomes from the full cohort of individuals with PPA showed significantly improved naming of trained items immediately post-treatment and at all follow-up assessments through 1 year. Generalized improvement on untrained items was significant up to 6 months post-treatment. The positive response to treatment was comparable regardless of session frequency or inclusion of a booster phase. Outcomes were comparable across PPA subtypes, as was maintenance of gains over the post-treatment period. Conclusion This study documents positive naming treatment outcomes for a group of individuals with PPA, demonstrating strong direct treatment effects, maintenance of gains up to 1 year post-treatment, and generalization to untrained items. Lexical retrieval treatment, in conjunction with daily home practice, had a strong positive effect that did not require more than 1 clinician-directed treatment session per week. Findings confirm that strategic training designed to capitalize on spared cognitive-linguistic abilities results in significant and lasting improvement, despite ongoing disease progression, in PPA.


Recent Advances in the Synthesis and Biological Activity of 8-Hydroxyquinolines.
Compounds containing the 8-hydroxyquinoline (8-HQ) 1 nucleus exhibit a wide range of biological activities, including antimicrobial, anticancer, and antifungal effects. The chemistry and biology of this group have attracted the attention of chemists, medicinal chemists, and professionals in health sciences. A number of prescribed drugs incorporate this group, and numerous 8-HQ- based molecules can be used to develop potent lead compounds with good efficacy and low toxicity. This review focusses on the recent advances in the synthesis of 8-HQ derivatives with different pharmacological properties, including anticancer, antiviral, and antibacterial activities. For this purpose, recent relevant references were searched in different known databases and search engines, such as MEDLINE (PubMed), Google Scholar, Science Direct, Scopus, Cochrane, Scientific Information Database (SID), SciFinder, and Institute for Scientific Information (ISI) Web of Knowledge. This review article provides a literature overview of the various synthetic strategies and biological activities of 8-HQ derivatives and covers the recent related literature. Taken together, compounds containing the 8-HQ moiety have huge therapeutic value and can act as potential building blocks for various pharmacologically active scaffolds. In addition, several described compounds in this review could act leads for the development of drugs against numerous diseases including cancer.


Developing a Web-based Tool Using Information and Communication Technologies to Expand the Reach and Impact of Photovoice.
Information and communication technologies are opening up vast new arenas for conducting the work of health promotion. Technology-based health promotions expand reach, standardize information and its delivery, provide opportunities for tailoring, create engaging interactivity within content delivery, provide for privacy and autonomy, improve portability, and lower delivery costs. This commentary describes the ongoing exploration and development of a web-based tool for enhancing the reach and impact of photovoice as a community change intervention. Features of the tool use information and communication technologies that integrate the use of an online learning management system, tailored messaging, gaming technology, interactive features, and the application of social media's power to increase the capacity of communities to employ comprehensive strategies to improve the health of their communities. It will enable individuals and groups to use photos and captions to assess the physical environment, social norms and behaviors of communities; raise community awareness of the factors contributing to ill-health in their communities, mobilize stakeholders, and inform environmental strategies and policy changes. We believe it will enhance the delivery of educational content about conducting photovoice projects, provide features unavailable without the application of information and communication technologies, and will be substantive advancement over existing photovoice resources.


L-arginine supplementation and risk factors of cardiovascular diseases in healthy men: a double-blind randomized clinical trial.
Context: The effect of L-arginine on risk factors of cardiovascular diseases (CVD) has mostly focused on western countries. Since cardiovascular diseases is the second cause of death in Iran and, as far as we are aware, there have been no studies about the effect of L-arginine on CVD risk factors, the aim of this trial was to assess the effects of L-arginine supplementation on CVD risk factors in healthy men. Objective: The purpose of this study was to evaluate the effect of low-dose L-arginine supplementation on CVD risk factors (lipid profile, blood sugar and blood pressure) in Iranian healthy men. Design, setting, participants: We conducted a double-blind randomized controlled trial in 56 patients selected from sport clubs at the Isfahan University of Medical Science between November 2013 and December 2013. Interventions: Healthy men received L-arginine supplementation (2000 mg daily) in the intervention group or placebo (2000 mg maltodextrin daily) in the control group for 45 days. Main outcome measure: The primary outcome measures were we measured the levels of fasting blood sugar, blood pressure and lipid profile including triglyceride (TG), cholesterol, LDL and HDL in healthy subjects. It was hypothesized that these measures would be significantly improved in those receiving L-arginine supplementation. at the beginning and end of the study. Results: In this trial, we had complete data for 52 healthy participants with mean age of 20.85±4.29 years. At the end of study, fasting blood sugar (P=0.001) and lipid profile (triglycerideTG (P<0.001), cholesterol (P<0.001), LDL (P=0.04), HDL (P=0.015)) decreased in the L-arginine group but we found no significant change in the placebo group. In addition, the reduction of fasting blood sugar and lipid profile in L-arginine was significant compared with placebo group. No significant changes were found about systolic (P=0.81) and diastolic blood pressure either in L-arginine or placebo group. (P=0.532). Conclusion: The use of L-arginine significantly improved outcomes compared to placebo.


A performance comparison of flat-panel imager-based MV and kV cone-beam CT.
The use of cone-beam computed tomography (CBCT) has been proposed for guiding the delivery of radiation therapy, and investigators have examined the use of both kilovoltage (kV) and megavoltage (MV) x-ray beams in the development of such CBCT systems. In this paper, the inherent contrast and signal-to-noise ratio (SNR) performance for a variety of existing and hypothetical detectors for CBCT are investigated analytically as a function of imaging dose and object size. Theoretical predictions are compared to the results of experimental investigations employing largearea flat-panel imagers (FPIs) at kV and MV energies. Measurements were performed on two different FPI-based CBCT systems: a bench-top prototype incorporating an FPI and kV x-ray source (100 kVp x rays), and a system incorporating an FPI mounted on the gantry of a medical linear accelerator (6 MV x rays). The SNR in volume reconstructions was measured as a function of dose and found to agree reasonably with theoretical predictions. These results confirm the theoretically predicted advantages of employing kV energy x rays in imaging soft-tissue structures found in the human body. While MV CBCT may provide a valuable means of correcting 3D setup errors and may offer an advantage in terms of simplicity of mechanical integration with a linear accelerator (e.g., implementation in place of a portal imager), kV CBCT offers significant performance advantages in terms of image contrast and SNR per unit dose for visualization of soft-tissue structures. The relatively poor SNR performance at MV energies is primarily a result of the low x-ray quantum efficiencies (approximately a few percent or less) that are currently achieved with FPIs at high energies. Furthermore, kV CBCT with an FPI offers the potential of combined volumetric and radiographic/fluoroscopic imaging using the same device.


Bioactive compounds in lettuce: Highlighting the benefits to human health and impacts of preharvest and postharvest practices.
Lettuce is one of the most commonly consumed leafy vegetables worldwide and is available throughout the entire year. Lettuce is also a significant source of natural phytochemicals. These compounds, including glycosylated flavonoids, phenolic acids, carotenoids, the vitamin B groups, ascorbic acid, tocopherols, and sesquiterpene lactones, are essential nutritional bioactive compounds. This review aims to provide a comprehensive understanding of the composition of health-promoting compounds in different types of lettuce, the potential health benefits of lettuce in reducing the risks of chronic diseases, and the effect of preharvest and postharvest practices on the biosynthesis and accumulation of health-promoting compounds in lettuce.


Tanshinone IIA Promotes M2 Microglia by ERβ/IL-10 Pathway and Attenuates Neuronal Loss in Mouse TBI Model.
Traumatic brain injury (TBI) is a major cause of morbidity and mortality worldwide. Increasing evidence indicates that activated microglia play an important role in the inflammatory response in TBI. Inhibiting M1 and stimulating M2 activated microglia have protective effects in several animal models of central nervous system (CNS) disorders. In the present study, we investigated whether tanshinone IIA (TNA) protects neurons by shifting microglia polarization in a mouse TBI model and further investigated the mechanism in vitro. Forty C57BL/6 mice were used to investigate the effect of TNA on microglia polarization in TBI. BV-2 cells were used to examine the mechanism of TNA in regulating microglia polarization. Normal saline (NS), TNA and the combination of TNA with ICI 182,780 (ICI, an estrogen receptor antagonist) were used to treat the TBI mice. After TBI, mice from each group demonstrated functional improvement. The improvement rate in mice treated with TNA was faster than other groups. ICI partially reversed the benefits from TNA treatment. TNA treatment significantly reduced TBI-induced neuronal loss. The number of microglia after TBI was not significantly changed by TNA treatment. However, TNA treatment significantly decreased M1 macrophage markers (iNOS, TNFα and IL-1β) and increased M2 macrophage markers (CD206, arginase 1 and Ym1). This effect was partially abolished by ICI. TNA treatment downregulated M1 macrophage markers and upregulated M2 macrophage markers in BV-2 cells under LPS stimulation. IL-10 was significantly increased by TNA treatment without a significantly change of IL-4 and IL-13 expression. IL-10 knockdown completely abolished the effect of TNA on microglial M2 polarization. Taken together, our data demonstrated that TNA attenuates neuronal loss in mouse TBI model and promotes M2 microglia by ERβ/IL-10 pathway. Thus, TNA could be a potential drug for TBI and/or the disorders that caused by microglial over-activation in CNS.


Synthesis and characterization of a novel fast-set proline-derivative-containing glass ionomer cement with enhanced mechanical properties.
In this study, a methacryloyl derivative of l-proline was synthesized, characterized and incorporated into a conventional glass ionomer cement (GIC) with a polyacid composition. Subsequently, the effects of incorporation of synthesized N-methacryloyl-proline and terpolymer on the GIC's mechanical and working properties were studied. 1-Methacryloylpyrrolidone-2-carboxylic acid was synthesized and used in a polymerization reaction with acrylic acid and itaconic acid in order to form terpolymer which was used in Fuji II commercial GIC formulations. Chemical structural characterization of the resulting products was performed using (1)H nuclear magnetic resonance and Fourier transform infrared spectroscopy. The viscosity and molecular weight of the terpolymer were also measured. The mechanical strength properties of the modified GICs were evaluated after 24h or 1 week of immersion in distilled water at 37 degrees C. Analysis of variance was used to study the statistical significance of the mechanical strengths and working properties, and to compare them with a control group. Results showed that N-methacryloyl-proline modified GICs exhibited significantly higher compressive strength (CS; 195-210MPa), higher diametral tensile strength (DTS; 19-26MPa) and higher biaxial flexural strength (38-46MPa) in comparison to Fuji II GIC (161-166MPa in CS, 12-14MPa in DTS and 13-18MPa in biaxial flexural strength). The working properties (setting and working time) of the modified samples showed that the modified cement was a fast-set cement. It was concluded that a novel amino acid-containing GIC has been developed in this study with 27%, 94% and 170% increases in values for compressive, diametral tensile and biaxial flexural strength, respectively, in comparison to commercial Fuji II GIC.


Electro-Responsive Surface Composition and Kinetics of an Ionic Liquid in a Polar Oil.
The quartz crystal microbalance (QCM) has been used to study how the interfacial layer of an ionic liquid dissolved in a polar oil at low weight percentages responds to changes in applied potential. The changes in surface composition at the QCM gold surface depend on both the magnitude and sign of the applied potential. The time-resolved response indicates that the relaxation kinetics are limited by the diffusion of ions in the interfacial region and not in the bulk, since there is no concentration dependence. The measured mass changes cannot be explained only in terms of simple ion exchange; the relative molecular volumes of the ions and the density changes in response to ion exclusion must be considered. The relaxation behavior of the potential between the electrodes upon disconnecting the applied potential is more complex than that observed for pure ionic liquids, but a measure of the surface charge can be extracted from the exponential decay when the rapid initial potential drop is accounted for. The adsorbed film at the gold surface consists predominantly of ionic liquid despite the low concentration, which is unsurprising given the surtactant-like structures of (some of) the ionic liquid ions. Changes in response to potential correspond to changes in the relative numbers of cations and anions, rather than a change in the oil composition. No evidence for an electric field induced change in viscosity is observed. This work shows conclusively that electric potentials can be used to control the surface composition, even in an oil-based system, and paves the way for other ion solvent studies.


Impact of human herpes virus 6 in liver transplantation.
Human herpes virus 6 (HHV-6) infects > 95% of humans. Primary infection which occurs mostly during the first 2 years of life in the form of roseola infantum, non-specific febrile illness, or an asymptomatic illness, results in latency. Reactivation of latent HHV-6 is common after liver transplantation. Since the majority of human beings harbor the latent virus, HHV-6 infections after liver transplantation are most probably caused by endogenous reactivation or superinfection. In a minority of cases, primary HHV-6 infection may occur when an HHV-6-seronegative individual receives a liver allograft from an HHV-6-seropositive donor. The vast majority of HHV-6 infections after liver transplantation are asymptomatic. Only in a minority of cases, when HHV-6 causes a febrile illness associated with rash and myelosuppression, hepatitis, gastroenteritis, pneumonitis, and encephalitis after liver transplantation. In addition, HHV-6 has been implicated in a variety of indirect effects, such as allograft rejection and increased predisposition to and severity of other infections, including cytomegalovirus, hepatitis C virus, and opportunistic fungi. Because of the uncommon nature of the clinical illnesses directly attributed to HHV-6, there is currently no recommended HHV-6-specific approach prevention after liver transplantation. Asymptomatic HHV-6 infection does not require antiviral treatment, while treatment of established HHV-6 disease is treated with intravenous ganciclovir, foscarnet, or cidofovir and this should be complemented by a reduction in immunosuppression.


Prolonged tau clearance and stress vulnerability rescue by pharmacological activation of autophagy in tauopathy neurons.
Tauopathies are neurodegenerative diseases associated with accumulation of abnormal tau protein in the brain. Patient iPSC-derived neuronal cell models replicate disease-relevant phenotypes ex vivo that can be pharmacologically targeted for drug discovery. Here, we explored autophagy as a mechanism to reduce tau burden in human neurons and, from a small-molecule screen, identify the mTOR inhibitors OSI-027, AZD2014 and AZD8055. These compounds are more potent than rapamycin, and robustly downregulate phosphorylated and insoluble tau, consequently reducing tau-mediated neuronal stress vulnerability. MTORC1 inhibition and autophagy activity are directly linked to tau clearance. Notably, single-dose treatment followed by washout leads to a prolonged reduction of tau levels and toxicity for 12 days, which is mirrored by a sustained effect on mTORC1 inhibition and autophagy. This new insight into the pharmacodynamics of mTOR inhibitors in regulation of neuronal autophagy may contribute to development of therapies for tauopathies.


Outcome of nonoperative management of displaced oblique spiral fractures of the fifth metatarsal shaft.
Nonoperative management has been the preferred treatment for displaced oblique spiral fractures of the fifth metatarsal shaft; yet a paucity of literature supports this claim. The purpose of this investigation was to report the incidence and long-term outcome in the largest cohort of these fractures reported to date. From 2006 through 2010, 2990 patients sustaining closed metatarsal fractures were seen and treated. Displaced, oblique, spiral fractures of the distal shaft of the fifth metatarsal were identified and follow-up was conducted. Only patients who were initially treated with nonoperative management were included. Patients were seen at 6 and 12 weeks, and a minimum 2-year follow-up was conducted. In addition, demographic information was obtained, and the Short Form-12 (SF-12) and Foot and Ankle Ability Measure (FAAM) were administered. Average follow-up was 3.5 years. In all, 142 acute fractures were managed for an incidence of 4.8% of all metatarsal fractures. There were 117 females and 25 males, average age was 55. FAAM activities of daily living subscale scores averaged 95.5 (±5.7), while FAAM sports subscales were 92.7 (±9.1). SF-12 physical and mental scores averaged 51.4 (±4.9) and 50.3 (±4.6), respectively. There were 2 delayed unions, 1 asymptomatic nonunion treated nonoperatively, and 2 painful nonunions that required open reduction internal fixation with bone grafting. This large cohort described the relative incidence and functional outcomes of displaced oblique fracture of shaft of the fifth metatarsal bone treated nonoperatively. Nonoperative management of these fractures resulted in excellent, long-term functional outcomes. Level II, prospective cohort study.


Diffusion tractography of the cervical spinal cord by using parallel imaging.
We applied a single-shot echo-planar axial diffusion tensor sequence with parallel imaging for tractography of the cervical cord. Placing a large region of interest on a reconstructed sagittal image, the cervical cord was depicted as a tract bundle showing color-encoded cephalocaudally. By setting one region of interest in the cerebral peduncle and another in the entire cord on source images, tracts on each side were visualized. This method is promising for tractography of the cervical cord.


Ring-Opening Dynamics of the Cyclopropyl Radical and Cation: the Transition State Nature of the Cyclopropyl Cation.
We provide compelling experimental and theoretical evidence for the transition state nature of the cyclopropyl cation. Synchrotron photoionization spectroscopy employing coincidence techniques together with a novel simulation based on high-accuracy ab initio calculations reveal that the cation is unstable via its allowed disrotatory ring-opening path. The ring strains of the cation and the radical are similar, but both ring opening paths for the radical are forbidden when the full electronic symmetries are considered. These findings are discussed in light of the early predictions by Longuet-Higgins alongside Woodward and Hoffman; we also propose a simple phase space explanation for the appearance of the cyclopropyl photoionization spectrum. The results of this work allow the refinement of the cyclopropane C-H bond dissociation energy, in addition to the cyclopropyl radical and cation cyclization energies, via the Active Thermochemical Tables approach.


Activity-based profiling of retaining β-glucosidases: a comparative study.
Activity-based protein profiling (ABPP) is a versatile strategy to report on enzyme activity in vitro, in situ, and in vivo. The development and use of ABPP tools and techniques has met with considerable success in monitoring physiological processes involving esterases and proteases. Activity-based profiling of glycosidases, on the other hand, has proven more difficult, and to date no broad-spectrum glycosidase activity-based probes (ABPs) have been reported. In a comparative study, we investigated both 2-deoxy-2-fluoroglycosides and cyclitol epoxides for their utility as a starting point towards retaining β-glucosidase ABP. We also investigated the merits of direct labeling and two-step bio-orthogonal labeling in reporting on glucosidase activity under various conditions. Our results demonstrate that 1) in general cyclitol epoxides are the superior glucosidase ABPs, 2) that direct labeling is the more efficient approach but it hinges on the ability of the glucosidase to be accommodated in the active site of the reporter (BODIPY) entity, and 3) that two-step bio-orthogonal labeling can be achieved on isolated enzymes but translating this protocol to cell extracts requires more investigation.


Biological mesh reconstruction of perineal wounds following enhanced abdominoperineal excision of rectum (APER).
An abdominoperineal excision of rectum (APER) may be required for rectal tumours less than 6 cm from the anal verge. Recently, the cylindrical APER has been used to prevent the "surgical waist" and so decrease margin involvement. However, removal of the levators leaves a large defect. Myocutaneous flaps [e.g. vertical rectus abdominis (VRAM)] are often used to fill the cylindrical resection defect, but have disadvantages associated with operative time, expertise and morbidity. We report our early experience of pelvic floor reconstruction with a biological mesh following cylindrical APER. Data on consecutive patients having cylindrical APER between January 2008 and November 2010 were collected. Outcomes were compared between a VRAM reconstruction group and a mesh group. In 15 consecutive patients with low rectal cancer, five patients had VRAM pelvic floor reconstruction prior to ten patients having biosynthetic mesh repairs. The median operative time for the VRAM cohort was 405 min, compared with 259 min for the mesh (p = 0.0013). The median length of postoperative stay was 20 days for VRAM and 10 days for the mesh group (p = 0.067). There were four early complications for the VRAM group compared with seven for the mesh cohort (p = 0.37). The median cost per patient for the VRAM cohort was £11,075 compared to a median cost of £6,513 for the Mesh (p = 0.0097). The use of a biological mesh for pelvic floor reconstruction following cylindrical APER is feasible with morbidity comparable to VRAM reconstruction. There is significant cost-saving using a biosynthetic mesh, mainly due to reduced length of stay.


Omega-3 PUFAs and vitamin D co-supplementation as a safe-effective therapeutic approach for core symptoms of autism spectrum disorder: case report and literature review.
Introduction: Autism spectrum disorder (ASD) is a group of neurodevelopmental disorders characterized by abnormal development of cognitive, social, and communicative skills. Although ASD aetiology and pathophysiology are still unclear, various nutritional factors have been investigated as potential risk factors for ASD development, including omega-3 polyunsaturated fatty acids (PUFAs) and vitamin D deficiency. In fact, both omega-3 PUFAs and vitamin D are important for brain development and function. Case report: Herein, we report the case of a 23-year-old young adult male with autism who was referred to our Unit due to a 12-month history of cyclic episodes of restlessness, agitation, irritability, oppositional and self-injurious behaviours. Laboratory tests documented a markedly altered omega-6/omega-3 balance, along with a vitamin D deficiency, as assessed by serum levels of 25-hydroxyvitamin D. Omega-3 and vitamin D co-supplementation was therefore started, with remarkable improvements in ASD symptoms throughout a 24-month follow-up period. A brief review of the literature for interventional studies evaluating the efficacy of omega-3 or vitamin D supplementation for the treatment of ASD-related symptoms is also provided. Conclusion: To our knowledge, this is the first case reporting remarkable beneficial effects on ASD symptoms deriving from omega-3 and vitamin D combination therapy. This case report suggests omega-3 and vitamin D co-supplementation as a potential safe-effective therapeutic strategy to treat core symptoms of ASD. However, larger studies are needed to evaluate the real efficacy of such therapeutic approach in a broader sample of ASD patients.


Spatial mapping of the biomechanical properties of the pericellular matrix of articular cartilage measured in situ via atomic force microscopy.
In articular cartilage, chondrocytes are surrounded by a narrow region called the pericellular matrix (PCM), which is biochemically, structurally, and mechanically distinct from the bulk extracellular matrix (ECM). Although multiple techniques have been used to measure the mechanical properties of the PCM using isolated chondrons (the PCM with enclosed cells), few studies have measured the biomechanical properties of the PCM in situ. The objective of this study was to quantify the in situ mechanical properties of the PCM and ECM of human, porcine, and murine articular cartilage using atomic force microscopy (AFM). Microscale elastic moduli were quantitatively measured for a region of interest using stiffness mapping, or force-volume mapping, via AFM. This technique was first validated by means of elastomeric models (polyacrylamide or polydimethylsiloxane) of a soft inclusion surrounded by a stiff medium. The elastic properties of the PCM were evaluated for regions surrounding cell voids in the middle/deep zone of sectioned articular cartilage samples. ECM elastic properties were evaluated in regions visually devoid of PCM. Stiffness mapping successfully depicted the spatial arrangement of moduli in both model and cartilage surfaces. The modulus of the PCM was significantly lower than that of the ECM in human, porcine, and murine articular cartilage, with a ratio of PCM to ECM properties of approximately 0.35 for all species. These findings are consistent with previous studies of mechanically isolated chondrons, and suggest that stiffness mapping via AFM can provide a means of determining microscale inhomogeneities in the mechanical properties of articular cartilage in situ.


Depression, anxiety and stress among patients with hematological malignancies and the association with quality of life: a cross-sectional study.
To evaluate psychological distress and quality of life of patients with hematological malignancies, as well as to identify the prognostic factors that aggravate their condition. A cross-sectional, descriptive study including 87 patients with Hematological Malignancies was conducted. Data were collected with an anonymous questionnaire consisted by A) a sheet with socio-demographic characteristics, B) the Depression, Stress and Stress Scale (DASS21), and C) World Health Organization Life Quality Scale (WHOQOL - BREF30). The processing and statistical analysis of the empirical material of the research were done using the SPSS (Statistical Package for Social Science) 22.0. The results showed that the age of the sample averaged 71.22 years (SD = 8.71). Regarding their marital status, single mothers were 9.2%, married 62.1%. With regard to disease-related features, 28.7% of patients had Hodgkin's Lymphoma, 27.6% Chronic Lymphocytic Leukemia, 27.6% MDL and 16.1% of patients had myelodysplastic syndrome Multiple myeloma. In the investigation of the relationship between the Mental health Scale and the Quality of Life, negative correlations of Depression, Stress, Stress and the total DAS Scale with all dimensions of Patient Quality of Life were found. Higher levels of psychological distress and poor mental health are associated with a low level of quality of life.


Studies on muscarinic binding sites in human brain identified with [3H]pirenzepine.
Pirenzepine, a potent antimuscarinic agent with apparent selectivity for a subtype (M1) of muscarinic receptors, was used in tritiated form to characterize its binding to human brain tissue. Specific [3H]pirenzepine binding showed rapid association and dissociation. From kinetic and competitive binding experiments, its KD was 5.5 nM and 9 nM, respectively. Regional distribution of [3H]pirenzepine binding determined in parallel with [3H]quinuclidinyl benzilate binding, a nonselective muscarinic antagonist, indicated a significant correlation for the maximum number of binding sites for the two radioligands in 13 brain regions, with the highest amount of binding for each in the putamen and the least in the cerebellum. Binding for [3H]pirenzepine averaged 57% of that for [3H]quinuclidinyl benzilate, with a range of 20% (cerebellum) to 77% (frontal cortex). Most antidepressants and neuroleptics tested had affinities for [3H]pirenzepine binding sites that were not significantly different from their previously reported values obtained with the use of [3H]quinuclidinyl benzilate.


[Two cases of syphilitic uveitis and HIV].
Case 1: HIV+ patient presenting unilateral panuveitis. Physical examination showed secondary signs of syphilis. The condition was treated with an excellent result. Case 2: Patient with bilateral panuveitis having received different treatments in another centre for a period of months. In the anamnesis we detected risk factors for AIDS and ordered appropriate tests to be done, with the patient resulting positive for HIV and syphilis. We proceeded to treat. The incidence of syphilis is rising in developed countries and its manifestations in HIV+ patients can be atypical. An adequate clinical history is fundamental for an early diagnosis. Macrolides can be an alternative treatment in penicillin-allergic patients.


Uses of the case-control and cohort epidemiological approaches in pediatric practice and research.
Increasing reliance is being placed on the use of quantitative epidemiological methods in the conduct and evaluation of pediatric research. The basic design features of two common types of observational studies, the case-control study and the cohort study, are reviewed. Advantages and disadvantages of these two study designs are discussed with emphasis on aspects such as the selection of comparison groups, avoiding selection and recall bias, gathering exposure information, controlling for potentially confounding factors, and methods of analysis. Appreciation of the salient features of these study design approaches should aid the clinician/researcher in the conduct of research endeavors as well as in critically reviewing the medical literature.


Examination of islets in the pancreas biopsy specimens from newly diagnosed type 1 (insulin-dependent) diabetic patients.
We attempted to examine the immunopathological change of the pancreatic islets of newly diagnosed Type 1 (insulin-dependent) diabetic patients and thereby to obtain useful information for the therapy of the patients. For this purpose, pancreas biopsy under laparoscopy was performed 2-4 months after the onset of Type 1 diabetes in seven newly diagnosed patients. All biopsies were performed safely without any complications. Immunohistochemical examination of the biopsy specimens revealed a marked decrease of insulin-containing cells, preservation of glucagon-containing cells, and various degrees of expression of MHC class I and class II antigens in islet cells and in endothelial cells within and around the islets. Signs of active autoimmune phenomena, e.g. lymphocytic infiltration or immunoglobulin deposition in islets, were not detected in any of these patients by light microscopical evaluation. We conclude that pancreas biopsy under laparoscopy has shown various immunological changes in the islets of newly diagnosed Type 1 diabetic patients. Pancreas biopsy, however, may not be suitable under the present protocol for the selection of patients for immunotherapy because of problems including sampling errors.


Antiproliferative, antioxidant and anti-inflammatory effects of hydroxytyrosol on human hepatoma HepG2 and Hep3B cell lines.
Olive oil intake has been shown to induce beneficial effect on health. This study aimed to investigate the effects of olive oil polyphenol hydroxytyrosol (HT) on cell proliferation and its antioxidant and anti-inflammatory capacity in human hepatoma Hep3B and HepG2 cell lines. Cell growth after HT treatment was measured by 3-(4,5 di-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test. Lipogenic enzyme activity was evaluated by radiochemical assay. Cell total antioxidant activity and cell interleukin-6 (IL-6) levels were measured by enyme linked immunosorbent assay (ELISA) methods. HT caused an evident antiproliferative effect mediated by inhibition of lipogenic enzymes. Moreover, HT induced activation of the cell antioxidant system and reduced cellular IL-6 levels. Our findings provide insights into the mechanisms of action of HT in the context of inhibition of cancer cell proliferation and prevention of oxidative stress in human hepatoma cells. Our results also show a down-regulation of lipogenic enzymes involved in cell proliferation.


[The role of interferons in neuroblastoma. 1: Antiproliferative effects].
Antiproliferative effects of interferon alpha, beta and gamma were investigated on several human neuroblastoma cell lines using the soft agar colony forming assay and the MTT-test. Investigations were carried out in order to prove whether there is any relationship between antiproliferative effects, inhibition of N-myc expression and the 2-5A system. Growth of neuroblastoma cells was inhibited by all three kinds of interferons in a concentration-dependent manner, however, rather high concentrations were necessary in some cell lines. Expression of N-myc oncogen was not inhibited by interferon-beta and no relationship between antiproliferative effects and the 2-5A system was observed. A vector containing a small N-myc fragment in antisense direction was constructed and transferred into the interferon insensitive human neuroblastoma cell line LS. After transformation, LS cells became sensitive to interferon beta: Proliferation as well as N-myc expression were inhibited and these processes are most probably associated with activation of the 2-5A system.


Porcelain color alteration after orthodontic bonding using three different surface preparation methods.
By increasing the number of adults seeking orthodontic treatment bonding orthodontic brackets to the surfaces other than intact enamel has become necessary. The purpose of this study was to evaluate the effect of three different surface preparation methods associated with orthodontic bonding on porcelain color alteration. In this in vitro study forty-five porcelain discs (6- mm diameter, 2- mm thickness) were fabricated. The color of the specimens was evaluated by means of a Vita Easyshade. Commision Internationale de I'Eclairage (CIE) L* a* b* system was used for color measurement. Then, the specimens were randomly divided into three groups (n = 15) with respect to the surface preparation methods including a 9.6% hydrofluoric acid (HF) + silane, sandblasting, and sandblasting + 9.6% HF + silane. Metal orthodontic brackets were bonded. Samples were stored in 37° c water for 24 hours. Afterward, the brackets were debonded with a debonding plier and porcelain surfaces were polished with a tungsten-carbide bur. The color assessment was done, and ΔE values were measured. ΔE = 3.7 units were considered as an acceptability threshold. Data were analyzed with Paired t-test and one-way ANOVA. Level of significance was set at P < 0.05. Orthodontic bonding changed the color parameters significantly. Mean L*, a* and b* difference were 1.35 ± 2.41, 0.19 ± 0.80, 0.89 ± 1.27 units, respectively (P = 0.003 for L*, P < 0.001 for a* and b*). There was not any significant difference in ΔE units between the groups (P = 0.456). In all the groups the mean ΔE values were below 3.7 units and within the clinically acceptable limit. Orthodontic treatment changed the CIE color parameters of porcelain surface. However, the color alteration is below the clinically acceptable threshold. With regard to color alteration, there is no difference between different surface conditioning methods.


Inclusion body myositis. A corticosteroid-resistant idiopathic inflammatory myopathy.
In seven patients with slowly progressive muscle weakness, inclusion body myositis (IBM) was diagnosed on biopsy. None had stigmata of collagen-vascular disease or malignancy. Serum creatine kinase levels were mildly or moderately increased. The six patients treated with prednisone did not improve. Needle electromyography showed a "myopathic" pattern in all patients, but four also had diffuse neurogenic changes with normal nerve conductions. Histologic study of muscle showed a mixture of small rounded fibers varying in size, atrophic angulated fibers forming small groups, and hypertrophic fibers. Variable amounts of inflammation, necrosis, and regeneration were seen in all specimens. All showed numerous intracytoplasmic vacuoles lined with purple-blue granules. Electron microscopy showed membranous whorls and masses of abnormal filaments measuring 14 to 18 nm in diameter. Although IBM seems to be a distinct type of inflammatory myopathy, its etiology and pathogenesis are not clear.


Identification of feeding and nutrition problems in young children with neuromotor involvement: a self-assessment.
The purpose of this article is to provide information needed to identify feeding and nutrition problems in children from birth to twenty-four months. Oral-motor and self-feeding skill development should be viewed within the framework of overall development and changing nutrition needs. Neuromotor dysfunction affects feeding and nutrition through changes in muscle tone, reflexes and the response to sensory stimulation. Nutrient compromises, including delays in texture progression, decreased fluid intake, and problems associated with self-feeding and food selection must be considered when assessing the nutrition/feeding needs of children. Successful completion of the self-assessment can be used to check the reader's basic understanding of the subject.


[Vitamin B 6 metabolism in phenylketonuria].
Based on the analysis of the dependence of 4-pyridoxic acid urinary excretion from pyridoxal-5'-phosphate blood plasma level and its statistic distribution it has been shown that vitamin B-6 metabolism in children suffering from phenylketonuria and therefore the criteria of the body saturation with this vitamin differ from those for healthy people. Increased pyridoxal-5'-phosphate blood plasma level has been demonstrated for PKU children. The concentration of 11 ng/ml should be considered as a bottom border of the adequate supply with vitamin B-6. The elevated vitamin B-2 intake approximates vitamin B-6 status indexes of sick children to those usually measured in healthy children. The necessity for the reevaluation of vitamin B-2 and B-6 optimal diet content under this disease and its biochemical validation are discussed.


The serological relationship between Brucella spp., Yersinia enterocolitica serotype IX and Salmonella serotypes of Kauffmann-White group N.
The serological relationship between Brucella spp., Yersinia enterocolitica IX, and the group N salmonella serotypes S. godesberg, S. landau, S. morehead, S. neusdorf, S. soerenga and S. urbana was examined using agglutination, antiglobulin, complement fixation, immunodiffusion and fluorescent antibody methods. Antisera to the group N salmonella serotypes all reacted to significant titres in agglutination and complement fixation, but not antiglobulin or immunodiffusion tests with smooth brucella antigens. These antisera also reacted in agglutination, but not antiglobulin, tests with Y. enterocolitica IX. They did not react significantly in any tests with rough brucella antigens. Conversely, antisera to smooth Brucella spp. agglutinated group N salmonellas to low titre and Y. enterocolitica IX to titres similar to those given against the homologous strain. Antiserum to Y. enterocolitica IX on the other hand reacted with smooth brucella antigens to high titre in agglutination, complement fixation and antiglobulin tests, and with the group N salmonella antigens to substantial titres in agglutination tests. In direct fluorescent antibody tests, smooth Brucella strains and Y. enterocolitica IX reacted strongly with FITC-labelled antibody to Br. abortus whereas the group N salmonella strains reacted weakly. In tests with monospecific antisera to the A and M determinants of Br. abortus and Br. melitensis respectively, Y. enterocolitica IX reacted only with the antiserum to the A determinant whereas group N salmonellas reacted to low titre with both A and M antisera. The results of cross-absorption tests confirmed this relationship and suggested that the O30 antigens of group N salmonella serotypes contained antigenic determinants similar to, but not identical with, the antigenic structure shared by smooth Brucella spp. and Y. enterocolitica IX.


[The status of lipid metabolism and free-radical processes in workers in a 30-kilometer exclusion zone].
Patterns were studied of lipid metabolism as was the state of free radical processes in different groups of those individuals working within a 30-km zone of estrangement depending on the length of time they stayed there. Atherogenically directed changes in blood lipid spectrum and processes of lipid peroxidation identified under working conditions in the zone of estrangement appeared to be more pronounced in subjects working at the object Ukryttia [correction of "Ukrytiye"] (Shelter) than in those in the zone working outside the object Ukryttia [correction of "Ukrytiye"].


A microfluidic device for quantifying bacterial chemotaxis in stable concentration gradients.
Chemotaxis allows bacteria to approach sources of attractant chemicals or to avoid sources of repellent chemicals. Bacteria constantly monitor the concentration of specific chemoeffectors by comparing the current concentration to the concentration detected a few seconds earlier. This comparison determines the net direction of movement. Although multiple, competing gradients often coexist in nature, conventional approaches for investigating bacterial chemotaxis are suboptimal for quantifying migration in response to concentration gradients of attractants and repellents. Here, we describe the development of a microfluidic chemotaxis model for presenting precise and stable concentration gradients of chemoeffectors to bacteria and quantitatively investigating their response to the applied gradient. The device is versatile in that concentration gradients of any desired absolute concentration and gradient strength can be easily generated by diffusive mixing. The device is demonstrated using the response of Escherichia coli RP437 to gradients of amino acids and nickel ions.


Handheld Active Add-On Control Unit for a Cable-Driven Flexible Endoscope.
The instruments currently used by surgeons for in utero treatment of the twin-to-twin transfusion syndrome (TTTS) are rigid or semi-rigid. Their poor dexterity makes this surgical intervention risky and the surgeon's work very complex. This paper proposes the design, assembly and quantitative evaluation of an add-on system intended to be placed on a commercialized cable-driven flexible endoscope. The add-on system is lightweight and easily exchangeable thanks to the McKibben muscle actuators embedded in its system. The combination of the flexible endoscope and the new add-on unit results in an easy controllable flexible instrument with great potential use in TTTS treatment, and especially for regions that are hard to reach with conventional instruments. The fetoscope has a precision of 7.4% over its entire bending range and allows to decrease the maximum planar force on the body wall of 6.15% compared to the original endoscope. The add-on control system also allows a more stable and precise actuation of the endoscope flexible tip.


Acute splenic sequestration crises in adults with sickle cell disease.
Reports of acute splenic sequestration crises in adults with sickle cell hemoglobin C disease or sickle cell thalassemia are rare, although an enlarged and distensible spleen persists in half of these patients. Seven episodes of acute splenic sequestration crises in four adults, two with sickle C disease and two with sickle thalassemia, are described. The crises were life-threatening and recurrent in all, but there were no fatalities. One patient had mild steady-state thrombocytopenia suggesting hypersplenism. Technetium 99m/sulfur colloid scanning of the spleen during the acute splenic sequestration crises in three patients showed almost total lack of splenic uptake or decreased uptake with intrasplenic filling defects thought to be splenic infarcts or hematomas on follow-up computed tomographic scanning. The scanning abnormalities resolved following recovery from the crises. Acute splenic sequestration crises probably are common in adults with sickle C disease and sickle thalassemia but may be underdiagnosed or misdiagnosed as splenic infarctions. The hematologic and splenic findings during acute splenic sequestration crises resemble those following splenic vein ligation in animals.


β-Catenin and SOX2 Interaction Regulate Visual Experience-Dependent Cell Homeostasis in the Developing Xenopus Thalamus.
In the vertebrate brain, sensory experience plays a crucial role in shaping thalamocortical connections for visual processing. However, it is still not clear how visual experience influences tissue homeostasis and neurogenesis in the developing thalamus. Here, we reported that the majority of SOX2-positive cells in the thalamus are differentiated neurons that receive visual inputs as early as stage 47 Xenopus. Visual deprivation (VD) for 2 days shifts the neurogenic balance toward proliferation at the expense of differentiation, which is accompanied by a reduction in nuclear-accumulated β-catenin in SOX2-positive neurons. The knockdown of β-catenin decreases the expression of SOX2 and increases the number of progenitor cells. Coimmunoprecipitation studies reveal the evolutionary conservation of strong interactions between β-catenin and SOX2. These findings indicate that β-catenin interacts with SOX2 to maintain homeostatic neurogenesis during thalamus development.


Lung cancer organoids analyzed on microwell arrays predict drug responses of patients within a week.
While the potential of patient-derived organoids (PDOs) to predict patients' responses to anti-cancer treatments has been well recognized, the lengthy time and the low efficiency in establishing PDOs hamper the implementation of PDO-based drug sensitivity tests in clinics. We first adapt a mechanical sample processing method to generate lung cancer organoids (LCOs) from surgically resected and biopsy tumor tissues. The LCOs recapitulate the histological and genetic features of the parental tumors and have the potential to expand indefinitely. By employing an integrated superhydrophobic microwell array chip (InSMAR-chip), we demonstrate hundreds of LCOs, a number that can be generated from most of the samples at passage 0, are sufficient to produce clinically meaningful drug responses within a week. The results prove our one-week drug tests are in good agreement with patient-derived xenografts, genetic mutations of tumors, and clinical outcomes. The LCO model coupled with the microwell device provides a technically feasible means for predicting patient-specific drug responses in clinical settings.


An evaluation of pre-poured selective media for the isolation of Neisseria gonorrhoeae.
Fourteen commercial media supplied as pre-poured plates were compared with an 'in-house' selective medium for their ability to support the growth of 105 gonococcal isolates (representing a wide variety of serovars encountered in natural infection), 25 meningococcal and 20 Neisseria lactamica isolates, and to inhibit the growth of 71 isolates of non-pathogenic neisseriae and miscellaneous organisms. Only two of the pre-poured plate media and the in-house selective medium yielded growth of duplicate cultures of all 105 gonococcal isolates after incubation for 24 h: one other medium provided growth of all the isolates after incubation for 48 h. The ability of the various media to suppress the growth of the 71 isolates of non-pathogenic neisseriae and miscellaneous organisms ranged from 97.2 to 71.8% of isolates inhibited. Of the four media that enabled growth of all the gonococcal strains, inhibition was 94.4% for the in-house medium, 85.9% and 80.3% for the two media on which all gonococci grew after 24 h and 71.8% for the medium on which all of the gonococci grew after 48 h. Failure of growth of gonococci was associated with: serogroup IA isolates (p<0.001), AHU auxotype (p<0.001) and the presence of vancomycin rather than lincomycin in the selective medium (p < 0.02). The use of 10% blood and a highly nutritious medium based on the original New York City (NYC) or modified New York City (MNYC) formulation were also important in supporting growth of gonococci. One of the main problems in lack of selectivity was a failure to inhibit the growth of yeasts. As effective inhibition of yeasts was obtained with other media containing the same concentration of amphotericin, failure may be due to batch variation of supplement, media preparation, or reduced shelf life of the media. None of the commercially available pre-poured media performed as well as the in-house medium despite the fact that some of the media were prepared to a very similar formula.


Nonoperative Treatment of Patellar Instability.
The treatment of patellar instability is challenging and typically begins with nonoperative methods. Clinical decisions are made on an individual basis and may vary according to a number of factors. First-time patellar dislocations most commonly occur during sports participation. Initial evaluation, including patient history, physical examination, and appropriate imaging, determines care. Although nonoperative treatment consists primarily of regressive immobilization and physical therapy, there is little evidence to support particular protocols for either. Factors that may contraindicate nonoperative treatment include osteochondral lesions and recurrent instability. In these cases, surgery is considered. Reported recurrence rates after nonoperative treatment of acute patellar dislocation are 15% to 44%. Well-designed studies are needed to determine not only the outcomes of nonoperative versus surgical treatment of patellar instability but also the risk factors that may predict poor outcomes in either group.


Feasibility of implementing the World Health Organization case management guideline for possible serious bacterial infection among young infants in Ntcheu district, Malawi.
Neonatal sepsis is a leading cause of mortality, yet the recommended inpatient treatment options are inaccessible to most families in low-income settings. In 2015, the World Health Organization released a guideline for outpatient treatment of young infants (0-59 days of age) with possible serious bacterial infection (PSBI) with simplified antibiotic regimens when referral was not feasible. If implemented widely, this guideline could prevent many deaths. Our implementation research evaluated the feasibility and acceptability of implementing the WHO guideline through the existing health system in Malawi. A prospective cohort study was conducted in 12 first-level health facilities in Ntcheu district. Trained health workers identified and treated young infants with PSBI signs with injection gentamicin for 2 days and oral amoxicillin for 7 days, whereas those with only fast breathing were treated with oral amoxicillin for 7 days. Health Surveillance Assistants (HSAs) were trained to promote care-seeking and to conduct home visits on day 3 and 6 to assess infants under treatment, encourage treatment adherence and remind the caregiver to return for facility follow up. Infants receiving outpatient treatment were followed up at health facility on day 4 and 8. The primary outcome was proportion of outpatient cases completing treatment per protocol. A total of 358 infants received outpatient treatment (202 clinical severe infection, 156 only fast breathing) from February to September 2017. Of these, 92.7% (332/358) met criteria for treatment completion and 88.8% (318/358) completed the day 4 follow-up. Twelve (3.4%) young infants clinically failed treatment with no reported deaths in those treated at outpatient level. This treatment failure rate was lower than those reported for the simplified regimens tested in the SATT (8-10%) and AFRINEST (5-8%) equivalency trials. More than half of infants (58.1%; 208/358) received HSA follow-up visits on days 3 and 6. Study results demonstrate the feasibility of outpatient treatment for sick young infants when referral is not feasible in Malawi, which will inform scale-up in other parts of Malawi and countries with similar health system constraints.


Facile difluoromethylation of aliphatic alcohols with an S-(difluoro-methyl)sulfonium salt: reaction, scope and mechanistic study.
A facile and practical approach for the difluoromethylation of aliphatic alcohols with an S-(difluoromethyl)sulfonium salt was developed. A wide variety of alcohols with broad functional groups are compatible to furnish the corresponding alkyl difluoromethyl ethers in good to excellent yields under mild reaction conditions. Control experiments and DFT computational studies suggest that the difluoromethylation of alcohols mainly proceeds via a difluorocarbene pathway involving a five-membered transition state with the participation of water, whose crucial role in this reaction was also elucidated by control experiments.


Degradation pathways of the commercial reactive azo dye Procion Red H-E7B under solar-assisted photo-Fenton reaction.
Reactive azo dye Procion Red H-E7B solutions have been submitted to solar-assisted photo-Fenton degradation. The solution color quickly disappears, indicating a fast degradation of the azo group. Nevertheless, complete DOC removal was not accomplished, in accordance with the presence of resistant triazine rings at the end of the reaction. The intermediates generated along the reaction time have been identified and quantified. LC-(ESI)-TOF-MS analysis allowed the detection of 18 aromatic compounds of different size and complexity. Some of them shared the same accurate mass, and consequently, the same empirical formula, but appeared at different chromatographic retention times, evidencing their different molecular structures. Heteroatom oxidation products like NH4+, NO3-, Cl-, and SO4(2-) have also been quantified and explanations of their release are proposed. Short chain carboxylic acids are also detected at long reaction times, as a previous step to complete dye mineralization. A link between the disappearance of the largest intermediate products and the increase of the solutions biodegradability has been established. Finally, taking into account all the findings of the present study and previous related works, the evolution from the original dye to the final products (triazine and CO2) is proposed in a general reaction scheme.


Mycobacterium tuberculosis ESAT6 induces IFN-β gene expression in Macrophages via TLRs-mediated signaling.
Mycobacterium tuberculosis is a highly virulent bacterium that causes tuberculosis. It infects about one third of the world's population. Type I interferons (IFNs) play a detrimental role in host defense against M. tuberculosis infection. Proteins secreted by M. tuberculosis through ESX-1 secretion system contribute to type I IFNs production. However, the precise mechanism by which 6-kDa early secretory antigen target (ESAT6), one of ESX-1-mediated secretory proteins, induces type I IFNs production in host cells is currently unclear. Therefore, the objective of the present study was to determine the underlying molecular mechanism regulating ESAT6-mediated gene expression of IFN-β in macrophages. Recombinant ESAT6 produced from E. coli expression system induced IFN-β gene expression in various types of macrophages such as mouse bone marrow-derived macrophages (BMDMs), peritoneal macrophages, and MH-S cells (murine alveolar macrophage cell line). Deficiency of TLR4 and TRIF absolutely abrogated ESAT6-induced IFN-β gene expression. TLR2 and MyD88 were partially involved in IFN-β gene expression in response to low dose of ESAT6. Another recombinant ESAT6 produced from baculovirus system also upregulated IFN-β gene expression via TLR4-dependent pathway. Polymyxin B (PMB) treatment impaired LPS-induced IFN-β expression. However, IFN-β expression induced by ESAT6 was not influenced by PMB. This suggests that ESAT6-mediated IFN-β expression is not due to LPS contamination. Treatment with ESAT6 resulted in activation of TBK1 and IRF3 in macrophages. Such activation was abolished in TLR4- and TRIF-deficient cells. Moreover, inhibition of IRF3 and TBK1 suppressed IFN-β gene expression in response to ESAT6. Our results suggest that ESAT6 might contribute to virulence of M. tuberculosis by regulating type I IFNs production through TLR4-TRIF signaling pathway.


Compliance with doxycycline therapy in sexually transmitted diseases clinics.
To determine rates of compliance with doxycycline therapy for patients attending two inner city sexually transmitted diseases (STD) clinics using the MEMS (Medication Event Monitoring System) technology (Aprex Corporation, Fremont, CA). An observational study. Two STD clinics in Brooklyn, New York and Birmingham, Alabama. Patients warranting doxycycline as antichlamydial therapy by usual clinical criteria (e.g., documented chlamydial infections, gonococcal urethritis, mucopurulent cervicitis) were enrolled consecutively from both clinics into four separate categories according to gender and the presence or absence of symptoms: symptomatic men (77), asymptomatic men (30), symptomatic women (83), asymptomatic women (33). In the clinic area, patients were given their doxycycline in standard 30-dram medication bottles fitted with the MEMS cap, which is capable of recording the date and time of each bottle opening and closing. This information was then retrieved using a software program developed by the manufacturer. Patients were instructed to return the bottle and cap at the completion of therapy. Efforts were made to contact those who did not return their bottles by both telephone and mail. Bottle openings as recorded by the MEMS were considered to represent use of medication. Patients were considered strictly compliant with prescription instructions if bottle openings and closings occurred at least twice daily for 6 consecutive days. Noncompliance was defined as initially opening the medication more than 48 hours after leaving the clinic or opening the bottle less than once daily for 5 consecutive days. Usage between these extremes was classified as intermediate. Eighty percent of 223 patients enrolled completed the study by returning their bottles. The rate of strict compliance with prescription instruction was 25%. The rate of noncompliance was 24%. Fifty-one percent used some intermediate amount of medication. There was no statistical difference in compliance by gender, presence or absence of symptoms, or site of enrollment. Few patients administered doxycycline in an STD clinic can be expected to take medication precisely as prescribed. Although most probably take enough to eradicate uncomplicated chlamydial infections, a sizable portion can be expected to use an inadequate amount of medication. This may contribute to persistence of genital chlamydia infections in the community.


Which patients benefit from physical activity on prescription (PAP)? A prospective observational analysis of factors that predict increased physical activity.
There is robust evidence that regular physical activity (PA) has positive health effects. However, the best PA methods and the most important correlates for promoting PA remain unclear. Physical activity on prescription (PAP) aims to increase the patient's motivation for and level of PA. This study investigated possible predictive baseline correlates associated with changes in the PA level over a 6-month period of PAP treatment in order to identify the primary care patients most likely to benefit from a PAP intervention. The study included 444 patients with metabolic risk factors who were aged 27 to 85 years and physically inactive. The patients received PAP treatment that included individual counseling plus an individually-tailored PA recommendation with a written prescription and individualised structured follow-up for 6 months. Eight baseline correlates of PA were analysed against the PA level at the 6-month follow-up in a predictor analysis. Five baseline correlates predicted the PA level at the 6-month follow-up: self-efficacy expectations for changing PA; the patient's preparedness and confidence regarding readiness to change PA; a BMI < 30; and a positive valued physical health. The proportion of patients increasing the PA level and achieving a PA level that was in accordance with public health recommendations was higher with a positive valued baseline correlate. The odds of achieving the recommended PA level increased substantially when 2 to 4 predictive correlates were present. PA levels increased to a greater extent among patients with low PA at baseline than patients with high PA at baseline, especially in combination with 2 to 4 positively-valued correlates (87-95% vs. 62-75%). This study identified potential predictive correlates of an increased PA level after a 6-month PAP intervention. This contributes to our understanding of PAP and could help individualise PAP support. The proportion of patients with the lowest PA level at baseline increased their PA level in a higher extent (84%) and thus may benefit the most from PAP. These results have clinical implications for behavioural change in those patients having the greatest health gains by increasing their PA level. ClinicalTrials.gov ; NCT03586011 . Retrospectively registered on July 17, 2018.


Multi-channel acoustic recording and automated analysis of Drosophila courtship songs.
Drosophila melanogaster has served as a powerful model system for genetic studies of courtship songs. To accelerate research on the genetic and neural mechanisms underlying courtship song, we have developed a sensitive recording system to simultaneously capture the acoustic signals from 32 separate pairs of courting flies as well as software for automated segmentation of songs. Our novel hardware design enables recording of low amplitude sounds in most laboratory environments. We demonstrate the power of this system by collecting, segmenting and analyzing over 18 hours of courtship song from 75 males from five wild-type strains of Drosophila melanogaster. Our analysis reveals previously undetected modulation of courtship song features and extensive natural genetic variation for most components of courtship song. Despite having a large dataset with sufficient power to detect subtle modulations of song, we were unable to identify previously reported periodic rhythms in the inter-pulse interval of song. We provide detailed instructions for assembling the hardware and for using our open-source segmentation software. Analysis of a large dataset of acoustic signals from Drosophila melanogaster provides novel insight into the structure and dynamics of species-specific courtship songs. Our new system for recording and analyzing fly acoustic signals should therefore greatly accelerate future studies of the genetics, neurobiology and evolution of courtship song.


Lateralization and functional organization of the locus coeruleus projection to the trigeminal somatosensory pathway in rat.
The primary goals of this study were to (1) examine the distribution of locus coeruleus (LC) neurons, which project to cortical and subcortical sites along the trigeminal somatosensory pathway in rats, and (2) determine the extent to which different regions within this ascending sensory system receive collateral projections from the same LC neuron. Long-Evans hooded rats received unilateral pressure injections of different combinations of retrograde fluorescent tracers into whisker-related regions of primary (SI) and secondary (SII) somatosensory cortices, the ventrobasal (VB) and posterior group (POm) nuclei of the thalamus, and the principalis nucleus of the trigeminal complex (PrV). Coronal sections (40-100 microm) through the LC were examined by fluorescence microscopy, and the distribution of retrogradely labeled cells was recorded. The major finding was that whisker-related regions of the cortex receive efferent projections from neurons concentrated in the caudal portion of the ipsilateral LC, whereas subcortical trigeminal somatosensory structures receive bilateral input from both LC nuclei. Despite the bilateral nature of the LC projection to subcortical sites, the majority of LC efferents to VB and POm thalamus originate in the ipsilateral LC nucleus, whereas projections to PrV originate primarily from the contralateral LC. An additional finding was that a relatively large proportion of LC cells, which project to a single somatosensory structure, also send axon collaterals to other relay sites along the same ascending somatosensory pathway. Taken together, these results suggest that the LC-noradrenergic system maintains a more selective relationship with functionally related efferent targets than has been previously appreciated.


Acting from the virtue of caring in nursing.
The author challenges the recently argued position of Helga Kuhse that caring is merely a preparatory stage to moral action and that impartial, principled thinking is required to make action moral, by suggesting a notion of caring as virtue. If caring is a virtue then acting from that virtue will be acting well. Acting from the virtue of caring involves eight features, which include not only that of being sensitive to, and concerned about, the patient, but also that of being aware of, and sensitive to, the relevant ethical principles. In this way, caring is seen as an overarching quality that gives action its moral character. The moral character of an action does not derive only from its having been performed in the light of principles.


Differential hormonal regulation of estrogen receptors ERalpha and ERbeta and androgen receptor expression in rat efferent ductules.
Estrogen receptors, in addition to the androgen receptor (AR), are expressed at high levels in efferent ductules of the male reproductive tract and it is now well recognized that estrogen receptor (ER) alpha is required for the maintenance of normal structure and function of the ductules. However, little is known regarding the hormonal regulation of the receptors themselves in the male. In the present study, efferent ductule ligation and castration, followed by replacement with testosterone, dihydro-testosterone (DHT) or estradiol was used to investigate the relative importance of circulating and luminal sources of steroid for the modulation of ERalpha, ERbeta and AR in rat efferent ductules. Uni- or bilateral castration and ligation did not affect the expression of ERalpha and ERbeta, but bilateral castration caused down-regulation of AR. Replacement with DHT and testosterone alone or in combination with estradiol caused the recovery of AR expression to control levels. A slight recovery of AR was also observed after estrogen replacement. ERalpha expression was decreased to nearly undetectable levels after estrogen replacement. On the other hand, ERbeta did not show evident effects following any of the treatments, suggesting a constitutive expression of this receptor. This differential modulation of the steroid hormone receptors highlights the importance of maintaining a physiological androgen-estrogen balance to regulate the structure and function of efferent ductules in the male.


Exploring the flavor life cycle of beers with varying alcohol content.
Decreasing alcohol in beer and increasing the pleasure of lower alcohol beers is a potential way to limit total alcohol consumption. Consumers' willingness to drink alcohol-reduced beers is mainly limited by unfavorable flavor characteristics that arise during consumption. To investigate the temporal flavor dominance during consumption, we analyzed the five most dominant beer flavors from nine different beers among three types of beer with varying alcohol content to assess the Flavor Life Cycle. Results show that beers with different alcohol content displayed similar flavor dominance (e.g., bitterness) and displayed differences in worty-off flavor, malty flavor, and astringency. In alcohol-free beers, worty-off flavor was most pronounced in dominating between 5 and 30 s and malty flavor increased after swallowing. For bitterness and astringency, higher alcohol content resulted in higher flavor dominance, especially prior to swallowing (≤40 sec). Based on these findings, we provide some brief advice to minimize unfavorable flavor experience during consumption of beer with lower alcohol. For now, consumers who want to enjoy beers with lower alcohol should consider flavor changes and focus on the favored and defocus on the less favored flavors.


Germline variants in HEY2 functional domains lead to congenital heart defects and thoracic aortic aneurysms.
In this study we aimed to establish the genetic cause of a myriad of cardiovascular defects prevalent in individuals from a genetically isolated population, who were found to share a common ancestor in 1728. Trio genome sequencing was carried out in an index patient with critical congenital heart disease (CHD); family members had either exome or Sanger sequencing. To confirm enrichment, we performed a gene-based association test and meta-analysis in two independent validation cohorts: one with 2685 CHD cases versus 4370 . These controls were also ancestry-matched (same as FTAA controls), and the other with 326 cases with familial thoracic aortic aneurysms (FTAA) and dissections versus 570 ancestry-matched controls. Functional consequences of identified variants were evaluated using expression studies. We identified a loss-of-function variant in the Notch target transcription factor-encoding gene HEY2. The homozygous state (n = 3) causes life-threatening congenital heart defects, while 80% of heterozygous carriers (n = 20) had cardiovascular defects, mainly CHD and FTAA of the ascending aorta. We confirm enrichment of rare risk variants in HEY2 functional domains after meta-analysis (MetaSKAT p = 0.018). Furthermore, we show that several identified variants lead to dysregulation of repression by HEY2. A homozygous germline loss-of-function variant in HEY2 leads to critical CHD. The majority of heterozygotes show a myriad of cardiovascular defects.


Normative Data for the 12-Item Sniffin' Sticks Odor Identification Test in Older Adults.
Quantitative olfactory assessment has demonstrated clinical utility for the evaluation of a range of neurologic, psychiatric, and sinonasal conditions. Here, we provide age, sex, race, and education-specific normative data for the 12-item Sniffin Sticks Odor Identification Test (SSOIT-12) in older Black and White U.S. adults without preclinical or clinical dementia or sinonasal disease. A sample of 2,224 Atherosclerosis Risk in Communities study participants aged 66-89 years were included. A normative regression equation was developed using a linear model for the association of age, sex, race, and education with odor identification score. Regression-based normative mean scores and percentiles were generated by age, sex, race, and education groups. Participants (mean age = 74 years, 59% women, 20% Black, 48% > high school education) had a mean SSOIT-12 score of 9.8. Age, sex, race, and education were all associated with odor identification performance (all ps < .05). A linear regression model for the predicted SSOIT-12 score was developed for use with an individual's actual SSOIT-12 score in order to calculate the Z-score and corresponding percentile for a specific age, sex, race, and education group. Data are also reported in tabular format. Our study provides SSOIT-12 normative data obtained from a large population of White and Black older adults without preclinical or clinical dementia or sinonasal disease living in the USA. These findings can aid clinicians in assessing the degree of olfactory loss, establishing concordance with a person's perception of olfactory difficulties and quantitatively monitoring changes in olfactory performance over time.


Identification of the sequences controlling cyclic AMP regulation and cell-type-specific expression of a prestalk-specific gene in Dictyostelium discoideum.
We have cloned and analyzed a developmentally and spatially regulated prestalk cell-specific gene from Dictyostelium discoideum. The gene encodes a protein highly homologous to the lysosomal cysteine proteinases cathepsin H and cathepsin B. Amino acid comparisons between these enzymes showed that the active-site amino acids were conserved, as were amino acids known to be important for catalysis and residues which form the intramolecular cysteine bridges. We have constructed a series of internal deletions, duplications, and linker scanner mutations within the region 300 base pairs 5' to the cap site. Analysis of expression of the mutations in transformants identified a approximately 35-base pair GC-rich region containing a dAdC/dGdT palindromic repeat and a G-rich box which is homologous to the 3' GT half of the palindromic repeat. Deletion or disruption of the G box resulted in a approximately 50-fold drop in the level of expression of the gene fusion in transformants in response to cyclic AMP in single-cell culture but did not affect the temporal pattern of regulation or control by cyclic AMP. The expression of such constructs during normal multicellular differentiation paralleled that of the endogenous gene; however, the level of RNA from the constructs was only approximately 10-fold lower than that of constructs containing the G box. Deletion of the 3' half of the palindromic sequence and the G box region resulted in a dramatic decrease in the level of transcription, although the constructs still showed proper temporal expression. These results suggest that this 35-base-pair region acts as an important part of the regulatory region for cell type and cyclic AMP regulation.


Advanced glycation end product-induced activation of NF-kappaB is suppressed by alpha-lipoic acid in cultured endothelial cells.
Depletion of cellular antioxidant defense mechanisms and the generation of oxygen free radicals by advanced glycation end products (AGEs) have been proposed to play a major role in the pathogenesis of diabetic vascular complications. Here we demonstrate that incubation of cultured bovine aortic endothelial cells (BAECs) with AGE albumin (500 nmol/l) resulted in the impairment of reduced glutathione (GSH) and ascorbic acid levels. As a consequence, increased cellular oxidative stress led to the activation of the transcription factor NF-kappaB and thus promoted the upregulation of various NF-kappaB-controlled genes, including endothelial tissue factor. Supplementation of the cellular antioxidative defense with the natural occurring antioxidant alpha-lipoic acid before AGE albumin induction completely prevented the AGE albumin-dependent depletion of reduced glutathione and ascorbic acid. Electrophoretic mobility shift assays (EMSAs) revealed that AGE albumin-mediated NF-kappaB activation was also reduced in a time- and dose-dependent manner as long as alpha-lipoic acid was added at least 30 min before AGE albumin stimulation. Inhibition was not due to physical interactions with protein DNA binding, since alpha-lipoic acid, directly included into the binding reaction, did not prevent binding activity of recombinant NF-kappaB. Western blots further demonstrated that alpha-lipoic acid inhibited the release and translocation of NF-kappaB from the cytoplasm into the nucleus. As a consequence, alpha-lipoic acid reduced AGE albumin-induced NF-kappaB mediated transcription and expression of endothelial genes relevant in diabetes, such as tissue factor and endothelin-1. Thus, supplementation of cellular antioxidative defense mechanisms by extracellularly administered alpha-lipoic acid reduces AGE albumin-induced endothelial dysfunction in vitro.


Proteome analysis reveals antiangiogenic environments in chronic wounds of diabetes mellitus type 2 patients.
In contrast to normal healing wounds, chronic wounds commonly show disturbances in proteins regulating wound healing processes, particularly those involved in cell proliferation and protein degradation. Multidimensional protein identification technology MS/MS was conducted to investigate and compare the protein composition of chronic diabetic foot exudates to exudates from split-skin donor sites of burn victims otherwise healthy. Spectral counting revealed 188 proteins differentially expressed (more than twofold and p-value <0.05) in chronic wounds. Most were involved in biological processes including inflammation, angiogenesis, and cell mortality. Increased expression of the inflammatory response stimulating S100 proteins, predominantly S100A8 and S100A9 (almost tenfold), was identified. Matrix metalloproteinases (MMPs) MMP1, MMP2, and MMP8 were identified to be elevated in chronic wounds with significant impact on collagen degradation and tissue destruction. Further, proteins with antiangiogenic properties were found at higher expression levels in chronic wounds. Reduced angiogenesis leads to drastic shortage in nutrition supply and causes increased cell death, demonstrated by Annexin A5 exclusively found in chronic wound exudates. However, excessive nucleic and cytosolic material infers cell death occurring not only by apoptosis but also by necrosis. In conclusion, mass spectrometric investigation of exudates from chronic wounds demonstrated dramatic impairment in wound repair with excessive inflammation, antiangiogenic environment, and accelerated cell death.


Beneficial Effects of Green Tea Catechins on Neurodegenerative Diseases.
Tea is one of the most consumed beverages in the world. Green tea, black tea, and oolong tea are made from the same plant Camellia sinensis (L.) O. Kuntze. Among them, green tea has been the most extensively studied for beneficial effects on diseases including cancer, obesity, diabetes, and inflammatory and neurodegenerative diseases. Several human observational and intervention studies have found beneficial effects of tea consumption on neurodegenerative impairment, such as cognitive dysfunction and memory loss. These studies supported the basis of tea's preventive effects of Parkinson's disease, but few studies have revealed such effects on Alzheimer's disease. In contrast, several human studies have not reported these favorable effects with regard to tea. This discrepancy may be due to incomplete adjustment of confounding factors, including the method of quantifying consumption, beverage temperature, cigarette smoking, alcohol consumption, and differences in genetic and environmental factors, such as race, sex, age, and lifestyle. Thus, more rigorous human studies are required to understand the neuroprotective effect of tea. A number of laboratory experiments demonstrated the benefits of green tea and green tea catechins (GTCs), such as epigallocatechin gallate (EGCG), and proposed action mechanisms. The targets of GTCs include the abnormal accumulation of fibrous proteins, such as Aβ and α-synuclein, inflammation, elevated expression of pro-apoptotic proteins, and oxidative stress, which are associated with neuronal cell dysfunction and death in the cerebral cortex. Computational molecular docking analysis revealed how EGCG can prevent the accumulation of fibrous proteins. These findings suggest that GTCs have the potential to be used in the prevention and treatment of neurodegenerative diseases and could be useful for the development of new drugs.


Capacity limitations of dishonesty.
Cognitive theories of dishonesty revolve around an automatic activation of honest response tendencies, which is assumed to impair response selection for the intended dishonest response. Clear-cut evidence for the claim is still limited, however. We therefore present a novel approach to dishonest responding that takes advantage of psychological refractory period methodology. Four experiments yielded evidence supporting the assumption of prolonged response selection during dishonest responding. Moreover, they also showed differences in early response activation and they revealed additional downstream consequences of this behavior that are currently not sufficiently covered by common theoretical models. Notably, these downstream consequences included increased monitoring relative to honest behavior. Our results thus provide an extensive coverage of the cognitive architecture of dishonest responses, informing current theorizing while simultaneously grounding the assumed processes in the framework of sensorimotor stage models of information processing. (PsycINFO Database Record (c) 2019 APA, all rights reserved).