A Novel Plasmonic Sensor Based on Dual-Channel D-Shaped Photonic Crystal Fiber for Enhanced Sensitivity in Simultaneous Detection of Different Analytes.
A dual-channel D-shaped photonic crystal fiber (PCF) based plasmonic sensor is proposed in this paper for the simultaneous detection of two different analytes using the surface plasmon resonance (SPR) technique. The sensor employs a 50 nm-thick layer of chemically stable gold on both cleaved surfaces of the PCF to induce the SPR effect. This configuration offers superior sensitivity and rapid response, making it highly effective for sensing applications. Numerical investigations are conducted using the finite element method (FEM). After optimizing the structural parameters, the sensor exhibits a maximum wavelength sensitivity of 10000 nm/RIU and an amplitude sensitivity of -216 RIU-1 between the two channels. Additionally, each channel of the sensor exhibits its unique maximal wavelength and amplitude sensitivities for different refractive index (RI) ranges. Both channels demonstrate a maximal wavelength sensitivity of 6000 nm/RIU. In the RI range of 1.31-1.41, Channel 1 (Ch1) and Channel 2 (Ch2) achieved their maximum amplitude sensitivities of -85.39RIU-1 and -304.52 RIU-1, respectively, with a resolution of 5×10-5. This sensor structure is noteworthy for its ability to measure both amplitude and wavelength sensitivity, providing enhanced performance characteristics suitable for various sensing purposes in chemical, biomedical, and industrial fields.


Purkinje cells from RyR2 mutant mice are highly arrhythmogenic but responsive to targeted therapy.
The Purkinje fiber network has been proposed as the source of arrhythmogenic Ca(2+) release events in catecholaminergic polymorphic ventricular tachycardia (CPVT), yet evidence supporting this mechanism at the cellular level is lacking. We sought to determine the frequency and severity of spontaneous Ca(2+) release events and the response to the antiarrhythmic agent flecainide in Purkinje cells and ventricular myocytes from RyR2(R4496C/+) CPVT mutant mice and littermate controls. We crossed RyR2(R4496C/+) knock-in mice with the newly described Cntn2-EGFP BAC transgenic mice, which express a fluorescent reporter gene in cells of the cardiac conduction system, including the distal Purkinje fiber network. Isolated ventricular myocytes (EGFP(-)) and Purkinje cells (EGFP(+)) from wild-type hearts and mutant hearts were distinguished by epifluorescence and intracellular Ca(2+) dynamics recorded by microfluorimetry. Both wild-type and RyR2(R4496C/+) mutant Purkinje cells displayed significantly slower kinetics of activation and relaxation compared to ventricular myocytes of the same genotype, and tau(decay) in the mutant Purkinje cells was significantly slower than that observed in wild-type Purkinje cells. Of the 4 groups studied, RyR2(R4496C/+) mutant Purkinje cells were also most likely to develop spontaneous Ca(2+) release events, and the number of events per cell was also significantly greater. Furthermore, with isoproterenol treatment, although all 4 groups showed increases in the frequency of arrhythmogenic Ca(2+(i)) events, the RyR2(R4496C/+) Purkinje cells responded with the most profound abnormalities in intracellular Ca(2+) handling, including a significant increase in the frequency of unstimulated Ca(2+(i)) events and the development of alternans, as well as isolated and sustained runs of triggered beats. Both Purkinje cells and ventricular myocytes from wild-type mice showed suppression of spontaneous Ca(2+) release events with flecainide, whereas in RyR2(R4496C/+) mice, the Purkinje cells were preferentially responsive to drug. In contrast, the RyR2 blocker tetracaine was equally efficacious in mutant Purkinje cells and ventricular myocytes. Purkinje cells display a greater propensity to develop abnormalities in intracellular Ca(2+) handling than ventricular myocytes. This proarrhythmic behavior is enhanced by disease-causing mutations in the RyR2 Ca(2+) release channel and greatly exacerbated by catecholaminergic stimulation, with the development of arrhythmogenic triggered beats. These data support the concept that Purkinje cells are critical contributors to arrhythmic triggers in animal models and humans with CPVT and suggest a broader role for the Purkinje fiber network in the genesis of ventricular arrhythmias.


[Sequential activation of various types of nuclear RNA polymerases of rat liver cells during the process of DNA synthesis induction by cycloheximide].
The dynamics of changes in the activity of RNA polymerases I and II (EC 18.104.22.168) and the activities of free and bound forms of RNA polymerase II in isolated rat liver nuclei after a singular injection of sublethal dose of cycloheximide (CHI) was studied. Activation of RNA polymerases occurs in three steps. The first step (1--2 hours after CHI injection) coincides with inhibition of protein synthesis and predominant activation of both forms of RNA polymerase II. The second step (6--24 hours after CHI injection) is characterized by de novo protein synthesis and activation of RNA polymerase I and RNA polymerase II. The third step (36--48 hours after CHI injection) witnesses activation of both RNA polymerases which coincides with the beginning of stimulation of DNA replication. It is assumed that this step is coupled with the synthesis of a special form of RNA polymerase, RNA polymerase IV, which is responsible for transcription of genes synthesizing various chromatin components (histones, NHB, RNA prime). At all steps the activity of free RNA polymerase II remains at a sufficiently high level. Consequently, the activity of chromatin is limited by chromatin structure rather than by RNA polymerase II content in it. Presumably, such activation of various forms of RNA polymerase is essential for cell preparation for DNA replication.


Personality factors of critical care nurses.
Two hundred members of the American Association of Critical-Care Nurses responded to a mail-out survey done to determine the psychologic profile of critical care nurses in terms of self-esteem, gender identity, and selected personality characteristics. The instruments used were Cattell's 16 PR, the Personal Attributes Questionnaire (PAQ), and the Texas Social Behavior Inventory (TSBI). Their personality factors tended to be aggressive, assertive, competitive, persevering, moralistic, resourceful, and mechanical. The nurses who enjoyed the field most were of the androgynous or masculine type and had high levels of self-esteem. On the basis of these findings, the nurse recruiter or faculty member doing career counseling could assess the personality characteristics, gender identity, and self-esteem levels of interested nurses. The goal would be to identify nurses who would both enjoy the field and remain active in critical care nursing after orientation. The goal could also be to help nurses dissatisfied with critical care nursing to seek means of improving their self-esteem.


A precursor-approach in constructing 3D ITO electrodes for the improved performance of photosystem I-cyt c photobioelectrodes.
In recent years the use of photoelectrodes based on conductive metal oxides has become very popular in the field of photovoltaics. The application of 3D electrodes holds great promise since they can integrate large amounts of photoactive proteins. In this study photosystem I (PSI) from the thermophilic cyanobacterium Thermosynechococcus elongatus was immobilized on 3D ITO electrodes and electrically wired via the redox protein cytochrome c (cyt c). The main goal, however, was the investigation of construction parameters of such electrodes for achieving a high performance. For this, ITO electrodes were constructed from liquid precursors resulting in improved transmission compared to previous nanoparticle-based preparation protocols. First, the doping level of Sn was varied for establishing suitable conditions for a fast cyt c electrochemistry on such 3D electrodes. In a second step the pore diameter was varied in order to elucidate optimal conditions. Third, the scalability of the template-based preparation was studied from 3 to 15 layers during spin coating and the subsequent baking step. In the thickness range from 3 to 17 μm no limitation in the protein immobilization and also in the photocurrent generation was found. Consequently, a photocurrent of about 270 μA cm-2 and a turnover number (Te) of 30 e- s-1 at PSI were achieved. Because of the high current flow the withdrawal of electrons at the stromal side of PSI becomes clearly rate limiting. Here improved transport conditions and alternative electron acceptors were studied to overcome this limitation.


[Prevalence of obesity and overweight in adolescents from Canary Islands, Spain. Relationship with breakfast and physical activity].
To estimate the prevalence of obesity and overweight in Canary adolescents, evaluating its association with breakfast intake and physical activity. Cross-sectional study of a representative sample of children aged 12-14 years living in the island of Gran Canaria. They were weighed and measured and the prevalence of overweight and obesity was determined according to the 85th and 97th percentiles of the Spanish body mass index tables. Breakfast and physical activity characteristics were also studied using questionnaires. The overall prevalence of obesity and overweight was 26.1%, higher in females (29.5%) than in males (22.8%). Obesity affects 14.8% of all teenagers (17.6% of girls and 12.0% of boys). Highest overweight and obesity levels affect those aged 12 years, decreasing progressively with age. Those boys who have a more complete breakfast have lower prevalence rates. There was no association between obesity and overweight with physical activity, as measured by the number of hours devoted to watching television or playing videogames as opposed to hours devoted to sport. The prevalence of overweight and obesity is high, especially in girls. We observed an inverse relationship between breakfast and its quality and obesity.


Gene activity during the early phase of androgen-stimulated rat prostate regrowth.
Androgenic steroids regulate the proliferation rate of normal and malignant prostate cells. In order to investigate the molecular basis of this control, we utilized Northern and Western blot techniques to measure changes in the expression of individual genes during the early phase of prostate regrowth. Vestigial ventral prostate glands of 7-day castrated rats showed increased numbers of replicating cells within 12 h of continuous pharmacological testosterone replacement as demonstrated by flow cytometric procedures. The period prior to the onset of proliferative enhancement was accompanied by the sequential induction of a variety of transcripts encoding gene products often associated with cell growth. Within 1 h of treatment, mature mRNA transcripts for c-fos were induced 6-fold, returning to control levels by 2 h. Other genes showed transiently elevated transcript levels after 2 h (c-Ha-ras, c-Ki-ras) or after 8 h (c-myc,c-myb, beta-actin, and Mr 70,000 heat shock protein) of testosterone replacement. Expression of the polypeptide encoded by c-Ha-ras was coordinately enhanced (2-fold) during this period, but not to the levels of the transcript (20-fold induction). Transcripts encoding basic fibroblast growth factor were increased 16 h and later, subsequent to the earlier enhancement in the proliferation rate. By placing these genes in a defined temporal order with regard to their expression in response to testosterone, we have constructed a map of gene activity during early prostate regrowth. This map shows a number of genes, the products of which might participate in the mechanism by which androgens induce mitogenesis of prostate cells.


State-ownership of tobacco industry: a 'fundamental conflict of interest' or a 'tremendous opportunity' for tobacco control?
Despite state-owned tobacco companies (SOTCs) accounting for over 40% of global production, the significance of state-ownership for tobacco control strategies has received limited academic and policy attention. The complex interests associated with SOTCs present diverse challenges for tobacco control policy, particularly in implementing Article 5.3 of WHO's Framework Convention on Tobacco Control (FCTC). Based on a review of existing literature, this paper examines current challenges and potential opportunities presented by governmental participation in the tobacco industry, identifying three contrasting perspectives from academic and policy sources. The first two perspectives centre on recognising that economic interests inherent in an SOTC are in tension with a government's public health responsibilities. This conflict can be perceived as either fundamental and fixed ('intrinsic conflict') or as amenable to either exacerbation or amelioration via organisational mechanisms ('institutionally-mediated conflict')-as suggested by the contrasting examples of China and Thailand. A third, less prominent perspective (which we refer to as 'interest alignment') suggests that it may be possible to radically alter the objectives and behaviour of SOTCs in order to advance tobacco control. Finally, we draw on this analysis to consider policy options for advancing tobacco control in countries with SOTCs. Guidance on implementation of Article 5.3 demonstrates strategic ambiguity by including elements of all three perspectives described above. We argue that legislative separation of tobacco control from SOTC oversight provides a desirable alternative to industry privatisation, and that radically realigning the goals of SOTCs to reduce tobacco consumption could make an important contribution to endgame strategies.


Applying Genomics and Proteomics in Translational Surgical Oncology Research.
Translational surgical research can range from investigating clinically oriented basic pathophysiological research via the correlation of molecular findings with a patient's clinical outcome to conducting treatment response studies. Herein, the specialist's opinion as a 'surgeon scientist' plays a pivotal role, e.g. in planning experimental setups focusing on the clinically most relevant needs. SUMMARY AND KEY MESSAGES: This review provides a survey of genomic and proteomic technologies and gives an up-to-date overview of those studies applying these technologies exemplarily in colorectal cancer-based biomarker research. Although current results are promising, future validation studies within multicenter networks are required to promote the translation of 'omics' from bench to bedside. In this endeavor, departments of surgery and institutes of experimental surgery together should play a fundamental role. The program for 'clinician scientists' recently launched by the German Research Society (DFG) could be one tool to promote interdisciplinary networks and surgeons' impact on translational research.


Health care system reform and public health. Protecting the safety net.
The most difficult to reach underserved populations are not likely to disappear even with universal insurance coverage. Universal coverage does offer all educated and motivated individuals, including those lacking financial resources, the opportunity to seek improved health through prevention services and treatment for illness. But, it would not guarantee that uneducated and unmotivated people, substance abusers, the chronically mentally ill, people with geographic or cultural barriers to access, or other traditionally underserved populations will use or be able to use a new system more wisely. These last groups, who, for various reasons, may underuse prevention services or delay contact until disease is transmitted or illness is advanced, will continue to add costs to a reformed health care system. Care must therefore be exercised in health care system reform to provide incentives for private-sector outreach to these problem populations. Even with incentives, however, history suggests that the private sector will either avoid or have difficulty incorporating the large groups of disadvantaged patients who in the past have relied on public health services and public hospitals for their health care. The public health sector will need to maintain the capacity to provide for the health care of many disadvantaged groups with historically poor access. It becomes critical in the budgetary trade-offs inherent in health care system reform that the infrastructure of the public health system not be damaged.(ABSTRACT TRUNCATED AT 250 WORDS)


Estimates of compliance with Canada's guidelines for low and moderate risk alcohol consumption: the importance of adjustment for underreporting in self-report surveys.
Alcohol is a leading cause of premature mortality; however, awareness of this and of some specific risks is low. Survey-based estimates of drinking at risk levels suffer from substantial underreporting. We show that alcohol use reported in the 2019 Canadian Alcohol and Drug Survey (CADS) accounted for just 38.06% of recorded alcohol consumption. This contributes to alcohol's risks being minimized, by researchers, the public, and policymakers. The new Canada's Guidance on Alcohol and Health (CGAH) defines "moderate risk" as 3 to 6 drinks/week for men and women. Employing published methods to correct for underreporting in the CADS, we estimate that in 2019 the proportion of drinkers at moderate risk of long-term harm to be 50.43% (up from 23.34% without adjustment). We further estimate that, collectively, these drinkers consumed 90.17% of all drinks consumed that year. Similarly, 92.82% of drinks were consumed on days when upper limits for short-term harm (2 drinks/day) were exceeded (up from 65.02% without adjustment). We conclude that adjustments for underreporting of alcohol use in Canada need to be incorporated routinely in public health monitoring. This might help mitigate the widespread underestimation of risky alcohol use as well as the neglect of this public health issue by policymakers.


The linear assembly of a pure glycoenzyme.
A labor of love: The synthesis of an active pure enzyme (RNase) glycoform by the native chemical ligation of a (glyco)peptide and peptide thioester fragments required numerous painstaking steps: the strategy was optimized, active inteins accessed, redox conditions fine-tuned, and glycoamino acid building blocks synthesized.


Dual role of phosphofructokinase-2/fructose bisphosphatase-2 in regulating the compartmentation and expression of glucokinase in hepatocytes.
Hepatic glucokinase is regulated by a 68-kDa regulatory protein (GKRP) that is both an inhibitor and nuclear receptor for glucokinase. We tested the role of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK2) in regulating glucokinase compartmentation in hepatocytes. PFK2 catalyzes formation or degradation of the regulator of glycolysis fructose 2,6-bisphosphate (fructose 2,6-P2), depending on its phosphorylation state (ser-32), and is also a glucokinase-binding protein. Incubation of hepatocytes at 25 mmol/l glucose causes translocation of glucokinase from the nucleus to the cytoplasm and an increase in fructose 2,6-P2. Glucagon caused phosphorylation of PFK2-ser-32, lowered the fructose 2,6-P2 concentration, and inhibited glucose-induced translocation of glucokinase. These effects of glucagon were reversed by expression of a kinase-active PFK2 mutant (S32A/H258A) that overrides the suppression of fructose 2,6-P2 but not by overexpression of wild-type PFK2. Overexpression of PFK2 potentiated glucokinase expression in hepatocytes transduced with an adenoviral vector-encoding glucokinase by a mechanism that does not involve stabilization of glucokinase protein from degradation. It is concluded that PFK2 has a dual role in regulating glucokinase in hepatocytes: it potentiates glucokinase protein expression by posttranscriptional mechanisms and favors its cytoplasmic compartmentation. Thus, it acts in a complementary mechanism to GKRP, which also regulates glucokinase protein expression and compartmentation.


Novel ophthalmic timolol meleate liposomal-hydrogel and its improved local glaucomatous therapeutic effect in vivo.
To overcome the limitations of common eye drops, the study developed a novel timolol mealate (TM) liposomal-hydrogel to enhance drug permeability and prolong residence time in the precorneal region, which achieved more effective local glaucomatous therapeutic effect. Firstly, TM liposome was prepared by an ammonium sulfate gradient-pH regulation method, which its entrapment efficiency reached up to 94% and its averaged particle size is 187 nm with narrow distribution. The corneal permeability through isolated rabbit cornea was measured by modified Franz-type diffusion cells. The results of trans-corneal penetration exhibited that the apparent permeability coefficients (P(app)) and the flow rates of steady state (J(ss)) of TM liposome was 1.50-fold higher than that of the commercialized eye drop, while TM liposome with 0.02% transcutol P was 2.19 times. In order to increase the retention time and improve the stability of liposome, we further developed a TM liposomal-hydrogel formulation by adding 1.0% HPMC K4M in TM liposome. The results showed an stability during a 120 days storage period than TM liposome. Precorneal retention study in vivo indicated that the optimal liposomal-hydrogel formulation had improved bioavailability and its retention time on rabbit corneal surface were significantly longer than that of pure liposomes or eye-drops. No obvious irritations to rabbit eyes were observed by histopathology microscopy after 7 days exposure.. Comparing to the eye drops, the TM liposomal-gel displayed prolonged therapeutic effect in cornea and greatly lowered the intraocular pressure IOP on the eyes of normal and glaucomatous pigmented rabbits.


[Geography of genetic processes in the population: genetic migration in Northern Eurasia (Central Asia region)].
Based on the data of the census of 1970, a map of the rates of gene migrations in the rural population of Central Asia and Kazakhstan was drawn. The corrected average value for 35 populations of oblasts (administrative regions) and autonomous republics was m = 0.0606 +/- 0.0102. The estimate obtained from the mapped data was 0.0799 +/- 0.0014, which is 1.2-fold higher than similar values for Europe. The relationship between the rate of gene migration m, the genetically effective population size Ne, and the density of elementary populations in the studied area Pp was analyzed by means of correlation and regression analyses. The following relationship was obtained: m = A/Ne + BPp, where A > 0 and B < 0. This was accounted for by the fact that the migration flow is mainly directed to the less densely populated rural areas. Given that the region was chosen somewhat artificially, studying gene migrations in Central Asia and Kazakhstan as separate historical, geographic, and genetic subregions of northern Eurasia might be a better approach.


Evaluation of a new reagent for anti-cytomegalovirus and anti-Epstein-Barr virus immunoglobulin G.
The Enzygnost alpha method was tested against the complement fixation test and anti-VCA immunofluorescence to determine the respective titers of anti-cytomegalovirus and anti-Epstein-Barr virus immunoglobulin G antibodies. For cytomegalovirus, the Enzygnost results showed 97.99% agreement with the readings obtained by the alternative method, with 100% sensitivity and 93.7% specificity. For Epstein-Barr virus, Enzygnost showed 97.71% agreement, 100% sensitivity, and 91.11% specificity.


Age-Predicted Maximal Heart Rate Equations Are Inaccurate for Use in Youth Male Soccer Players.
The purpose of this study was to compare the differences between measured (MHRobt) and predicted (MHRpred) maximal heart rate (MHR) in youth athletes. In total, 30 male soccer players [14.6 (0.6) y] volunteered to participate in this study. MHRobt was determined via maximal-effort graded exercise test. Age-predicted MHR (MHRpred) was calculated for each participant using equations by Fox, Tanaka, Shargal, and Nikolaidis. Mean differences were compared using Friedman's 2-way analysis of variance and post hoc pairwise comparisons. Agreement between MHRobt and MHRpred values was calculated using the Bland-Altman method. There were no significant differences between MHRobt and MHRpred from the Fox (P = .777) and Nikolaidis (P = .037) equations. The Tanaka and Shargal equations significantly underestimated MHRobt (P < .001). All 4 equations produced 95% limits of agreement of ±15.0 beats per minute around the constant error. The results show that the Fox and Nikolaidis equations produced the smallest mean difference in predicting MHRobt. However, the wide limits of agreement suggests that none of the equations adequately account for individual variability in MHRobt. Practitioners should avoid applying these equations in youth athletes and utilize a lab or field testing protocol to obtain MHR.


Forty years trends in timing of pubertal growth spurt in 157,000 Danish school children.
Entering puberty is an important milestone in reproductive life and secular changes in the timing of puberty may be an important indicator of the general reproductive health in a population. Too early puberty is associated with several psychosocial and health problems. The aim of our study was to determine if the age at onset of pubertal growth spurt (OGS) and at peak height velocity (PHV) during puberty show secular trends during four decades in a large cohort of school children. Annual measurements of height were available in all children born from 1930 to 1969 who attended primary school in the Copenhagen Municipality. 135,223 girls and 21,612 boys fulfilled the criteria for determining age at OGS and age at PHV. These physiological events were used as markers of pubertal development in our computerized method in order to evaluate any secular trends in pubertal maturation during the study period (year of birth 1930 to 1969). In this period, age at OGS declined statistically significantly by 0.2 and 0.4 years in girls and boys, respectively, whereas age at PHV declined statistically significantly by 0.5 and 0.3 years in girls and boys, respectively. The decline was non-linear with a levelling off in the children born between 1940 and 1955. The duration of puberty, as defined by the difference between age at OGS and age at PHV, increased slightly in boys, whereas it decreased in girls. Our finding of declining age at OGS and at PHV indicates a secular trend towards earlier sexual maturation of Danish children born between 1930 and 1969. Only minor changes were observed in duration of puberty assessed by the difference in ages at OGS and PHV.


Sympathetic activation of splenic T-lymphocytes in hypertension of adult offspring programmed by maternal high fructose exposure.
Whereas neuroimmune crosstalk between the sympathetic nervous system (SNS) and immune cells in the pathophysiology of hypertension is recognized, the exact effect of SNS on T-lymphocyte in hypertension remains controversial. This study assessed the hypothesis that excitation of the SNS activates splenic T-lymphocytes through redox signaling, leading to the production of pro-inflammatory cytokines and the development of hypertension. Status of T-lymphocyte activation, reactive oxygen species (ROS) production and pro-inflammatory cytokines expression in the spleen were examined in a rodent model of hypertension programmed by maternal high fructose diet (HFD) exposure. Maternal HFD exposure enhanced SNS activity and activated both CD4+ and CD8+ T-lymphocytes in the spleen of young offspring, compared to age-matched offspring exposed to maternal normal diet (ND). Maternal HFD exposure also induced tissue oxidative stress and expression of pro-inflammatory cytokines in the spleen of HFD offspring. All those cellular and molecular events were ameliorated following splenic nerve denervation (SND) by thermoablation. In contrast, activation of splenic sympathetic nerve by nicotine treatment resulted in the enhancement of tissue ROS level and activation of CD4+ and CD8+ T-cells in the spleen of ND offspring; these molecular events were attenuated by treatment with a ROS scavenger, tempol. Finally, the increase in systolic blood pressure (SBP) programmed in adult offspring by maternal HFD exposure was diminished by SND, whereas activation of splenic sympathetic nerve increased basal SBP in young ND offspring. These findings suggest that excitation of the SNS may activate splenic T-lymphocytes, leading to hypertension programming in adult offspring induced by maternal HFD exposure. Moreover, tissue oxidative stress induced by the splenic sympathetic overactivation may serve as a mediator that couples the neuroimmune crosstalk to prime programmed hypertension in HFD offspring.


Differential effects of delay on time-based prospective memory in younger and older adults.
The current study measured forgetting in a time-based, naturalistic prospective memory (PM) task. In Experiment 1, younger and older participants were asked to mail a stamped postcard on a date that was delayed 1, 2, 5, 14, or 28 days in the future. In Experiment 2, a different sample of older participants completed the same task with similar delays to replicate results for the older sample in Experiment 1. Overall, older participants were more likely than younger participants to mail the postcard on time. In addition, delay affected on-time return rates more for the younger participants than the older participants. Younger participants' return rates illustrated the typical forgetting curve seen in numerous retrospective memory studies (i.e., rapid decline at shorter delays and slower decline for longer delays). However, older participants' return rates only declined at the longest delays. These results indicate that time-based PM performance declines with an increase in delay, but the form of the decline may differ across age groups.


Interactions of macromolecules with the mammalian cell surface.
The characterisation of fluoresceinphosphatidylethanolamine (FPE) as a real-time indicator of the electrostatic nature of the cell membrane surface is described. The conditions appropriate for the labelling of cell membranes and the implementation of FPE as a tool to monitor the interactions of various proteins and peptides with membranes are outlined. Some complications attributed to the erythrocyte glycocalyx are examined. In addition it is shown using neuraminidase as an example, that some types of enzyme-catalysed reactions on the cell surface may be monitored in real time. It is also shown that information concerning the binding of several proteins such as serum albumin and monoclonal antibodies are accessible with this technique. The albumin in particular is shown to exhibit a saturation of binding, the analysis of which indicates that the dissociation constant for erythrocytes was determined to be 8 microM and for lymphocytes to be almost 3 microM. On the basis of this comparison together with artificial membranes, the membrane protein components of the lymphocyte surface are implicated in the binding of albumin or the erythrocyte membrane proteins reduce the affinity of the cell surface for albumin.


Deficit or Difference? Effects of Altered Auditory Feedback on Speech Fluency and Kinematic Variability in Adults Who Stutter.
Purpose The purpose of this study was to test whether adults who stutter (AWS) display a different range of sensitivity to delayed auditory feedback (DAF). Two experiments were conducted to assess the fluency of AWS under long-latency DAF and to test the effect of short-latency DAF on speech kinematic variability in AWS. Method In Experiment 1, 15 AWS performed a conversational speaking task under nonaltered auditory feedback and 250-ms DAF. The rates of stuttering-like disfluencies, other disfluencies, and speech errors and articulation rate were compared. In Experiment 2, 13 AWS and 15 adults who do not stutter (AWNS) read three utterances under four auditory feedback conditions: nonaltered auditory feedback, amplified auditory feedback, 25-ms DAF, and 50-ms DAF. Across-utterance kinematic variability (spatiotemporal index) and within-utterance variability (percent determinism and stability) were compared between groups. Results In Experiment 1, under 250-ms DAF, the rate of stuttering-like disfluencies and speech errors increased significantly, while articulation rate decreased significantly in AWS. In Experiment 2, AWS exhibited higher kinematic variability than AWNS across the feedback conditions. Under 25-ms DAF, the spatiotemporal index of AWS decreased significantly compared to the other feedback conditions. AWS showed lower overall percent determinism than AWNS, but their percent determinism increased under 50-ms DAF to approximate that of AWNS. Conclusions Auditory feedback manipulations can alter speech fluency and kinematic variability in AWS. Longer latency auditory feedback delays induce speech disruptions, while subtle auditory feedback manipulations potentially benefit speech motor control. Both AWS and AWNS are susceptible to auditory feedback during speech production, but AWS appear to exhibit a distinct continuum of sensitivity.


Crystalline, mixed-valence manganese analogue of prussian blue: magnetic, spectroscopic, X-ray and neutron diffraction studies.
The compound of stoichiometry Mn(II)3[Mn(III)(CN)6]2.zH2O (z = 12-16) (1) forms air-stable, transparent red crystals. Low-temperature single crystal optical spectroscopy and single crystal X-ray diffraction provide compelling evidence for N-bonded high-spin manganese(II), and C-bonded low-spin manganese(III) ions arranged in a disordered, face-centered cubic lattice analogous to that of Prussian Blue. X-ray and neutron diffraction show structured diffuse scattering indicative of partially correlated (rather than random) substitutions of [Mn(III)(CN)6] ions by (H2O)6 clusters. Magnetic susceptibility measurements and elastic neutron scattering experiments indicate a ferrimagnetic structure below the critical temperature Tc = 35.5 K.


Gamification of dietary decision-making in an elementary-school cafeteria.
Despite the known health benefits of doing so, most US children do not consume enough fruits and vegetables (FV). School-based interventions can be effective in increasing FV consumption, but the most effective of these require that schools allocate their time, effort, and financial resources to implementing the program: expenditures that schools may be reluctant to provide in climates of academic accountability and economic austerity. The present demonstration project used a behaviorally based gamification approach to develop an intervention designed to increase FV consumption while minimizing material and labor costs to the school. During the intervention, the school (N = 180 students in grades K-8) played a cooperative game in which school-level goals were met by consuming higher-than-normal amounts of either fruit or vegetables (alternating-treatments experimental design). School-level consumption was quantified using a weight-based waste measure in the cafeteria. Over a period of 13 school days, fruit consumption increased by 66% and vegetable consumption by 44% above baseline levels. Use of an alternating-treatment time-series design with differential levels of FV consumption on days when fruit or vegetable was targeted for improvement supported the role of the intervention in these overall consumption increases. In post-intervention surveys, teachers rated the intervention as practical in the classroom and enjoyed by their students. Parent surveys revealed that children were more willing to try new FV at home and increased their consumption of FV following the intervention. These findings suggest that a behaviorally based gamification approach may prove practically useful in addressing concerns about poor dietary decision-making by children in schools.


Tetramethylpyrazine improves spatial cognitive impairment induced by permanent occlusion of bilateral common carotid arteries or scopolamine in rats.
Effects of tetramethylpyrazine (TMP), a major constituent of Ligusticum chuanxiong, on spatial cognitive impairment induced by permanent occlusion of bilateral common carotid arteries (2VO) and scopolamine were investigated using 8-arm radial maze performance in rats. Permanent 2VO produced a severe learning deficit in non-pretrained rats. Daily administration of TMP (3-10 mg/kg, i.p.) from the 3rd day after permanent 2VO significantly improved the learning deficit. TMP did not influence the impairment of the retention task in the pretrained permanent 2VO rats, but it tended to reduce the number of errors elevated by 3-min delay interposition in these rats. In the scopolamine model, scopolamine (0.3 mg/kg, i.p.) significantly decreased the initial correct response and increased the number of errors. Single administration of TMP (1-3 mg/kg, i.p.) dose-dependently reversed the scopolamine-induced impairment of the maze performance. These results suggest that TMP has therapeutic potential for the treatment of dementia caused by cholinergic dysfunction and/or decrease of cerebral blood flow.


Germ line stem cell competition in postnatal mouse testes.
Niche is believed to affect stem cell behavior. In self-renewing systems for which functional transplantation assays are available, it has long been assumed that stem cells are fixed in the niche and that ablative treatments to remove endogenous stem cells are required for successful donor engraftment. Our results demonstrate that enriched populations of donor stem cells can produce long-lasting spermatogenic colonies in testes of immature and mature, nonablated mice, albeit at a lower frequency than in ablated mice. Colonization of nonablated recipient testes by neonate, pup, and cryptorchid adult donor spermatogonial stem cells demonstrates that competition for niche begins soon after birth and that endogenous stem cells influence the degree and pattern of donor cell colonization. Thus, a dynamic relationship between stem cell and niche exists in the testis, as has been suggested for hematopoiesis. Therefore, similar competitive properties of donor stem cells may be characteristic of all self-renewing systems.


Hairy cell leukemia: a clinical review based on 725 cases of the Italian Cooperative Group (ICGHCL). Italian Cooperative Group for Hairy Cell Leukemia.
The Italian Registry for hairy cell leukemia (HCL) has recorded 725 patients with HCL diagnosed over 25 years. We analysed this large series of patients with the aim of providing an evaluation of changes in clinical presentation, impact of initial therapy and modifications in prognostic factors over the period of two decades. Over time, a progressive down-staging of the disease at the onset, along with a reduction of patients with severe anemia and marked splenomegaly, has been observed. A second malignancy was found in 3.7% of patients, mostly detected several years after the onset of HCL. A striking improvement of survival rates has been observed, from 58.9% survival at five years for patients diagnosed before 1985 to 87.5% at five years for patients diagnosed after 1985 (p < 0.0001). Before 1985 hemoglobin alone provided prognostic information, whereas after 1985, clinical stage and the number of leukocytes correlated better with patient outcome. Survivals at 5 and 10 years were 34.4% and 29.6% respectively for untreated patients, 58.8% and 44.1% for patients receiving chemotherapy, steroids or other drugs, 64.1% and 56.1% for splenectomized patients and 88.9% (at 5 years) for alpha interferon (IFN)-treated patients (p < 0.0001). Our findings suggest that IFN has improved the prognosis of HCL, and that it must be considered a good initial treatment for patients with HCL.


Evaluation of three different techniques used to measure chest wall movements in children.
Several techniques exist for assessing chest wall movements in children, that incorporate different measurement principles. Although the output from such devices appears very similar, the comparability of data from different devices needs to be evaluated. We have investigated the simultaneous measurement of chest wall movement using three different devices in 12 children recovering from intensive care. The devices used were inductance plethysmography (assessing thoracic cross-sectional area), magnetometers (assessing thoracic diameter), and a Hall device strain guage (assessing thoracic circumference). Measurements of respiratory timing and of phase angle between rib cage and abdomen in these patients showed a close agreement between devices in ventilated patients. However, occasional inconsistencies occurred in patients who were breathing spontaneously. We suggest that it may not always be appropriate to directly compare data on chest wall movements in children recorded using different measurement techniques. The best method of measurement varies with the clinical picture.


Function of formate dehydrogenases in Desulfovibrio vulgaris Hildenborough energy metabolism.
The genome of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough encodes three formate dehydrogenases (FDHs), two of which are soluble periplasmic enzymes (FdhAB and FdhABC3) and one that is periplasmic but membrane-associated (FdhM). FdhAB and FdhABC3 were recently shown to be the main enzymes present during growth with lactate, formate or hydrogen. To address the role of these two enzymes, ΔfdhAB and ΔfdhABC3, mutants were generated and studied. Different phenotypes were observed in the presence of either molybdenum or tungsten, since both enzymes were important for growth on formate in the presence of Mo, whereas in the presence of W only FdhAB played a role. Both ΔfdhAB and ΔfdhABC3 mutants displayed defects in growth with lactate and sulfate providing the first direct evidence for the involvement of formate cycling under these conditions. In support of this mechanism, incubation of concentrated cell suspensions of the mutant strains with lactate and limiting sulfate also gave elevated formate concentrations, as compared to the wild-type strain. In contrast, both mutants grew similarly to the wild-type with H2 and sulfate. In the absence of sulfate, the wild-type D. vulgaris cells produced formate when supplied with H2 and CO2, which resulted from CO2 reduction by the periplasmic FDHs. The conversion of H2 and CO2 to formate allows the reversible storage of reducing power in a much more soluble molecule. Furthermore, we propose this may be an expression of the ability of some sulfate-reducing bacteria to grow by hydrogen oxidation, in syntrophy with organisms that consume formate, but are less efficient in H2 utilization.


Rapid detection of Streptococcus uberis in raw milk by loop-mediated isothermal amplification.
A loop-mediated isothermal amplification (LAMP) method to detect Streptococcus uberis in raw milk was developed and evaluated. Three genes (sodA, pauA, cpn60) were assessed for their suitability as targets in LAMP. The analytical sensitivity was 120, 120, and 12 fg per assay for the sodA, pauA, and cpn60 assays, respectively, with a detectable signal within 8 min for the highest concentration (12ng/assay) and ~60 min for the lowest concentrations. The LAMP assays correctly identified 7 Strep. uberis strains among a set of 83 mastitis pathogens. To enable DNA isolation from raw milk, a new method was used in which a pretreatment with a cocktail of lysing enzymes was performed before an established procedure. This method resulted in an analytical sensitivity of 48 cfu/assay for the sodA LAMP assay using raw milk spiked with Strep. uberis, corresponding to 2.4×10(4) cfu/mL milk. For raw milk samples from cows experimentally infected with Strep. uberis, results of enumeration were largely reflected by results of LAMP. Evaluation of the sodA LAMP assay with 100 raw milk field samples, of which 50 were Strep. uberis culture-negative and 50 Strep. uberis culture-positive, showed that the assay had a diagnostic sensitivity of 96.0% and a diagnostic specificity of 96.0%. In conclusion, the described LAMP assay may offer a simple alternative for convenient and sensitive detection of S. uberis in raw milk, provided a compatible rapid DNA isolation procedure is available.


Renal cell carcinoma with TFE3 translocation and succinate dehydrogenase B mutation.
Translocation renal cell carcinoma and succinate dehydrogenase (SDH)-deficient renal cell carcinoma are now recognized as specific renal tumor types in the World Health Organization (WHO) classification. Both have limited immunohistochemical positivity for epithelial markers, and the spectrum of morphology continues to widen for both of these entities. We identified four renal cell carcinomas with positive TFE3 immunohistochemical staining and negative SDHB staining. The patients (2F, 2M) ranged in age from 19 to 65 years. All tumors were composed, at least in part, of eosinophilic cells. Cytoplasmic inclusions, prominent nucleoli, and mitotic figures were seen in three tumors. Psammoma bodies were also present in two tumors. Using immunohistochemistry, a broad spectrum of commonly used renal tumor markers yielded nonspecific, limited positivity, including uniformly positive reactions for PAX8 but negative results for cathepsin K and HMB45. Fluorescence in situ hybridization results showed the presence of TFE3 gene rearrangement in all four tumors, and molecular analysis revealed SDHB mutations in neoplastic cells of three tumors. In one case, the same SDHB mutation was confirmed in the adjacent non-neoplastic tissue. We report for the first time the presence of both TFE3 translocation and SDHB mutation in the same tumor.


Endothelial Barrier Integrity Is Disrupted In Vitro by Heme and by Serum From Sickle Cell Disease Patients.
Free extracellular heme has been shown to activate several compartments of innate immunity, acting as a danger-associated molecular pattern (DAMP) in hemolytic diseases. Although localized endothelial barrier (EB) disruption is an important part of inflammation that allows circulating leukocytes to reach inflamed tissues, non-localized/deregulated disruption of the EB can lead to widespread microvascular hyperpermeability and secondary tissue damage. In mouse models of sickle cell disease (SCD), EB disruption has been associated with the development of a form of acute lung injury that closely resembles acute chest syndrome (ACS), and that can be elicited by acute heme infusion. Here we explored the effect of heme on EB integrity using human endothelial cell monolayers, in experimental conditions that include elements that more closely resemble in vivo conditions. EB integrity was assessed by electric cell-substrate impedance sensing in the presence of varying concentrations of heme and sera from SCD patients or healthy volunteers. Heme caused a dose-dependent decrease of the electrical resistance of cell monolayers, consistent with EB disruption, which was confirmed by staining of junction protein VE-cadherin. In addition, sera from SCD patients, but not from healthy volunteers, were also capable to induce EB disruption. Interestingly, these effects were not associated with total heme levels in serum. However, when heme was added to sera from SCD patients, but not from healthy volunteers, EB disruption could be elicited, and this effect was associated with hemopexin serum levels. Together our in vitro studies provide additional support to the concept of heme as a DAMP in hemolytic conditions.


Combining magnetoencephalography with magnetic resonance imaging enhances learning of surrogate-biomarkers.
Electrophysiological methods, that is M/EEG, provide unique views into brain health. Yet, when building predictive models from brain data, it is often unclear how electrophysiology should be combined with other neuroimaging methods. Information can be redundant, useful common representations of multimodal data may not be obvious and multimodal data collection can be medically contraindicated, which reduces applicability. Here, we propose a multimodal model to robustly combine MEG, MRI and fMRI for prediction. We focus on age prediction as a surrogate biomarker in 674 subjects from the Cam-CAN dataset. Strikingly, MEG, fMRI and MRI showed additive effects supporting distinct brain-behavior associations. Moreover, the contribution of MEG was best explained by cortical power spectra between 8 and 30 Hz. Finally, we demonstrate that the model preserves benefits of stacking when some data is missing. The proposed framework, hence, enables multimodal learning for a wide range of biomarkers from diverse types of brain signals.


Assessment of the levels of nitric oxide (NO) and cytokines (IL-5, IL-6, IL-13, TNF, IFN-gamma) in giardiosis.
The current study aims to determine the involvement of cellular responses in combating Giardia intestinalis invasion. The study group consisted of 44 women and 18 men, aged 18-72 years, infected with G. intestinalis. The diagnosis was established based on laboratory investigations (examination of stool, choloscopy, GSA-65). Blood for analysis was collected before antiparasitic treatment and two weeks after treatment termination. The control group consisted of 22 women and 18 men aged 20-45 years. The serum concentrations of IL-5, IL-6, IL-13, TNF, IFN-g were assayed using a set of Quantikine human. The concentrations of NO in the serum were determined using a set of Total Nitric Oxide Assay. Patients infected with G. intestinalis showed a statistically significant increase in the levels of NO, IFN-g and IL-13. Even the antiparasitic treatment applied did not reduce the levels of these parameters and only caused a rise in IL-6. Our study showed a lack of acute inflammatory state in the course of G. intestinalis infection.


Indications for rapid evolution of trait means and thermal plasticity in range-expanding populations of a butterfly.
Currently, poleward range expansions are observed in many taxa, often in response to anthropogenic climate change. At the expanding front, populations likely face cooler and more variable temperature conditions, imposing thermal selection. This may result in changes in trait means or plasticity, the relative contribution of which is not well understood. We, here, investigate evolutionary change in range-expanding populations of the butterfly Pieris mannii, by comparing populations from the core and the newly established northern range under laboratory conditions. We observed both changes in trait means and in thermal reaction norms. Range-expanding populations showed a more rapid development, potentially indicative of counter-gradient variation and an increased cold tolerance compared with core populations. Genotype-environment interactions prevailed in all associated traits, such that the above differences were restricted to cooler environmental conditions. In range-expanding populations, plasticity was decreased in developmental traits enabling relatively rapid growth even under cooler conditions but increased in cold tolerance arguably promoting higher activity under thermally challenging conditions. Notably, these changes must have occurred within a time period of ca. 10 years only. Our results suggest, in line with contemporary theory, that the evolution of plasticity may play a hitherto underestimated role for adaptation to climatic variation. However, rather than generally increased or decreased levels of plasticity, our results indicate fine-tuned, trait-specific evolutionary responses to increase fitness in novel environments.


Experiences of living with juvenile idiopathic arthritis: a qualitative systematic review protocol.
The objective of this review is to identify, critically appraise and synthesize the available qualitative evidence to understand the experiences of children, young adults and their carers living with juvenile idiopathic arthritis in any setting. Juvenile idiopathic arthritis is the most common rheumatic disease in childhood. Despite the availability of effective treatments, the disease still has negative impacts on patients' and carers' lives. Patients' and carers' experiences of living with juvenile idiopathic arthritis have been recognized as important in the measurement of health status and treatment implementation. Addressing these needs will facilitate more effective management and treatment of the disease. This protocol describes a method for a systematic review regarding the perspectives from patients and carers in order to highlight the needs of families throughout their juvenile idiopathic arthritis journey. Studies on the experiences of patients aged <21 years who have been diagnosed with juvenile idiopathic arthritis according to the International League of Associations for Rheumatology criteria, as well as the experiences of their carers, will be considered. Papers included in this review will include, but not be limited to, designs such as phenomenology, grounded theory and ethnography. A comprehensive search using PubMed, CINAHL, Embase, PsycINFO and Web of Science was undertaken in August 2019. Available studies published in English from 2001 to 2019 will be included. The recommended JBI method for study selection, critical appraisal, data extraction and data synthesis will be used. PROSPERO (CRD42019133165).


Leukemia inhibitory factor blocks expression of Crx and Nrl transcription factors to inhibit photoreceptor differentiation.
Activating ligands of gp130, including leukemia inhibitory factor (LIF), can block differentiation and function of retinal neurons. This study focused on determining whether LIF inhibits differentiation of photoreceptors by altering cell fate or by blocking the expression of essential transcription factors in vivo. Transgenic mice were generated that had lens-specific expression of the secreted human LIF protein. Retinal differentiation was assessed by histology and by gene expression analysis, with in situ hybridization, immunohistochemistry, and real-time qRT-PCR. Electroretinograms were used to assess retinal function. LIF did not prevent or alter the timing of outer and inner nuclear layer separation, but it inhibited phototransduction gene expression in both rods and cones, thereby blocking functional maturation of photoreceptors. LIF also reduced the expression of Crx, Nrl, and Nr2e3, and upregulated the expression of transcription inhibitors Baf and Fiz1. LIF expression did not appear to alter photoreceptor cell fate specification, but it inhibited subsequent differentiation. These results suggest that gp130 ligands can inhibit photoreceptor functional differentiation by reducing Crx- and Nrl-dependent transcription.


Alterations in blood glucose and plasma glucagon concentrations during deep brain stimulation in the shell region of the nucleus accumbens in rats.
Deep brain stimulation (DBS) of the nucleus accumbens (NAc) is an effective therapy for obsessive compulsive disorder (OCD) and is currently under investigation as a treatment for eating disorders. DBS of this area is associated with altered food intake and pharmacological treatment of OCD is associated with the risk of developing type 2 diabetes. Therefore we examined if DBS of the NAc-shell (sNAc) influences glucose metabolism. Male Wistar rats were subjected to DBS, or sham stimulation, for a period of 1 h. To assess the effects of stimulation on blood glucose and glucoregulatory hormones, blood samples were drawn before, during and after stimulation. Subsequently, all animals were used for quantitative assessment of Fos immunoreactivity in the lateral hypothalamic area (LHA) using computerized image analysis. DBS of the sNAc rapidly increased plasma concentrations of glucagon and glucose while sham stimulation and DBS outside the sNAc were ineffective. In addition, the increase in glucose was dependent on DBS intensity. In contrast, the DBS-induced increase in plasma corticosterone concentrations was independent of intensity and region, indicating that the observed DBS-induced metabolic changes were not due to corticosterone release. Stimulation of the sNAc with 200 μA increased Fos immunoreactivity in the LHA compared to sham or 100 μA stimulated animals. These data show that DBS of the sNAc alters glucose metabolism in a region- and intensity- dependent manner in association with neuronal activation in the LHA. Moreover, these data illustrate the need to monitor changes in glucose metabolism during DBS-treatment of OCD patients.


Activated lactoferrin's ability to inhibit Candida growth and block yeast adhesion to the vaginal epithelial monolayer.
To study in vitro growth-inhibitory effects of activated lactoferrin (ALF) against vaginal isolates of Candida species and to measure the ability of ALF to block interactions of Candida albicans and Candida glabrata to the vaginal epithelial (VE) monolayer. In vitro effects of ALF on growth of C albicans and C glabrata in Sabouraud dextrose (SD) broth were measured as change in broth turbidity by microscale optical density assay. ALF was tested at 5 and 2.5 mg/mL concentrations against 105 yeast cell inoculum at 370 degrees C for 96 hours and compared with native lactoferrin and control (growth in broth without ALF). VE cells were isolated from human vaginal tissue biopsies to establish a functional monolayer for yeast interaction studies. ALF effects on Candida interactions with the VE monolayer were tested using 3H-thymidine-labeled yeast. Prophylactic (treatment prior to yeast inoculation onto VE) and therapeutic (treatment to detach VE-adherent yeast) potential of ALF (5 mg/mL) was evaluated against vaginal isolates of C albicans strain NTRL809A and C glabrata strain NTRL131G. Growth of Candida species indicated that a 105 yeast inoculum in SD broth proliferated to a stationary growth equilibrium (approximately 10(9) yeast cell density) in 18 hours (approximately 2 hours of generation time). ALF (5 mg/mL) elicited >96 hours of total stasis (100% growth inhibition) and was significantly effective against both Candida species (p < 0.0001). At 2.5 mg/mL dilution, ALF sustained total stasis activity to an average of 18 hours and 24 hours for C albicans (n = 5) and C glabrata (n = 5), respectively. Interaction studies indicated avid binding of C albicans (70 - 140 x 10(3) yeast) and C glabrata (50 - 75 x 10(3) yeast) per square centimeter of VE monolayer. ALF-treated VE showed significant blockade (p < 0.05) of yeast adhesion by 33% and 58% with C albicans and C glabrata, respectively. ALF treatment of yeast-VE complexes resulted in significant detachment (p < 0.05) of C albicans and C glabrata, by 58% and 51%, respectively. ALF is a natural fungistatic agent with potent yeast adhesion-blocking and detachment properties and is effective against the vaginal pathogens C albicans and C glabrata.


DNA methylation and genome rearrangement characteristics of phase change in cultured shoots of Sequoia sempervirens.
Epigenetic machinery regulates the expression of individual genes and plays a crucial role in globally shaping and maintaining developmental patterning. We studied the extent of DNA methylation in the nucleus, mitochondrion and chloroplast in cultured Sequoia sempervirens (coast redwood) adult, juvenile and rejuvenated shoots by measuring the ratio of methylcytosine to total cytosine using high-performance liquid chromatography (HPLC). We also analyzed nuclear DNA (nuDNA) polymorphisms of different shoot types by methylation-sensitive amplified fragment length polymorphism (MSAP) and Southern blot analysis. The extent of nuDNA methylation was greater in the adult vegetative than juvenile and rejuvenated shoots (8% vs 6.5-7.5%). In contrast, the proportion of methylcytosine was higher in mitochondrial DNA (mDNA) of juvenile and rejuvenated shoots than adult shoots (6.6% vs 7.8-8.2%). MSAP and Southern blot analyses identified three MSAP fragments which could be applied as phase-specific molecular markers. We also found nuclear genome and mtDNA rearrangement may be as important as DNA methylation status during the phase change. Our findings strongly suggest that DNA methylation and genome rearrangement may affect the dynamic tissue- and cell type-specific changes that determine the developmental phase of S. sempervirens shoots.


Adipose-Derived Mesenchymal Stem Cell Exosomes Suppress Hepatocellular Carcinoma Growth in a Rat Model: Apparent Diffusion Coefficient, Natural Killer T-Cell Responses, and Histopathological Features.
We sought to evaluate the effects of adipose-derived mesenchymal stem cells (ADMSCs) exosomes on hepatocellular carcinoma (HCC) in rats using apparent diffusion coefficient (ADC), natural killer T-cell (NKT-cell) responses, and histopathological features. ADMSC-derived exosomes appeared as nanoparticles (30-90 nm) on electron microscopy and were positive for CD63, tumor susceptibility gene-101, and β-catenin on western blotting. The control (n = 8) and exosome-treated (n = 8) rats with N1S1-induced HCC underwent baseline and posttreatment day 10 and day 20 magnetic resonance imaging and measurement of ADC. Magnetic resonance imaging showed rapidly enlarged HCCs with low ADCs in the controls. The exosome-treated rats showed partial but nonsignificant tumor reduction, and significant ADC and ADC ratio increases on day 10. On day 20, the exosome-treated rats harbored significantly smaller tumors and volume ratios, higher ADC and ADC ratios, more circulating and intratumoral NKT-cells, and low-grade HCC (P < 0.05 for all comparisons) compared to the controls. The ADC and volume ratios exhibited significant inverse correlations (P < 0.001, R (2) = 0.679). ADMSC-derived exosomes promoted NKT-cell antitumor responses in rats, thereby facilitating HCC suppression, early ADC increase, and low-grade tumor differentiation. ADC may be an early biomarker of treatment response.


Metabolic fate of glutamate carbon in rat renal tubules. Studies with 13C nuclear magnetic resonance and gas chromatography-mass spectrometry.
13C-n.m.r. spectroscopy and g.c.-m.s. were used to determine the metabolic fate of glutamate carbon in rat kidney. The main purpose was to characterize the effect of chronic metabolic acidosis on the utilization of glutamate carbon. Renal tubules obtained from normal and chronically acidotic rats were incubated in Krebs buffer, pH 7.4, in the presence of 2.5 mM-[3-13C]glutamate. During the course of incubation the concentrations of total glucose and NH3 were significantly (P less than 0.05) higher in tissue from acidotic rats. The levels of some tricarboxylic-acid-cycle intermediates were higher (P less than 0.05) in control tissue. In control tissue, 13C-n.m.r. spectra demonstrated a significantly higher rate of 13C appearance of aspartate, glutamine and [2,4-13C]glutamate. However, in acidosis the resonances of [13C]glucose carbon atoms were significantly higher. In the control, approx. 15% of glutamate carbon was accounted for by [13C]glucose formation as against 30% in chronic acidosis. However, in control tissue, 44% of glutamate carbon utilization was accounted for by recycling to glutamate and formation of aspartate, glutamine and GABA. In acidosis, only 11% was so recovered. Analysis of 15NH3 formation during the course of incubation with 2.5 mM-[15N]glutamate demonstrated a positive association between the appearance of [13C]glucose and 15NH3 both in the control and in acidosis. The data suggest that the control of gluconeogenesis and ammoniagenesis in acidosis is, in part, referable to a diminution in the rate of the reductive amination of alpha-oxoglutarate, that of the transamination reaction and that of glutamine synthesis.


[Cytotoxic action of active forms of oxygen and mechanisms of development of a chronic process in the liver in liver disease].
The paper deals with the damaging action of active oxygen forms (AOF) on the cell, which is associated with impairments of membranous structures and their functional properties. Based on the results of their own investigations, the authors forward a concept of the stereotypic pattern of changes in the enzyme mechanisms of AOF utilization in chronic hepatic diseases of viral etiology. It is suggested that impaired hepatocytic AOF formation and inactivation may be metabolic mechanisms mediating the status and development of a chronic process in the liver in its viral damage. Promising approaches to treating oxidative impairments are also discussed.


The Association of Perfluoroalkyl Substance Exposure and a Serum Liver Function Marker in Korean Adults.
Perfluoroalkyl substances (PFAS), widely used throughout industry and daily life, are currently one of the environmental pollutants garnering the most attention worldwide. Recently, environmental pollutants have had a high profile as one of the main causes of chronic liver disease, such as non-alcoholic fatty liver disease. Research on PFAS is actively underway. Although Korea has a remarkably high prevalence of chronic liver disease, and it continues to increase, only a few studies have revealed the relationship between PFAS and liver disease. In addition, regulations on PFAS in Korea are delayed compared to developed countries, such as Europe and the United States, and public interest is insufficient compared to others. Therefore, we would like to investigate the exposure of Koreans to PFAS in the blood and examine the relationship between these substances and markers of liver function (AST, ALT, and GGT). This study was based on the results of the Korean National Environmental Health Survey (KoNEHS) 2015-2017 (Cycle 3), and a total of 2961 subjects were selected. The concentration of PFAS in the blood of Korean adults was measured to be significantly higher based on the geometric mean compared to the results of recently investigated American adults based on the National Health and Nutrition Examination Survey (NHANES, 2017-2018). A multivariable linear regression analysis adjusted for age, sex, body mass index (BMI), smoking status, alcohol intake, and regular exercise was performed to examine changes in three liver function markers as the serum PFAS concentration increased. We found that some of the five PFAS (PFOA, PFOS, PFHxS, PFNA, and PFDeA) were significantly associated with increased liver enzymes. It is necessary to recognize the threat of PFAS to the human body and to discuss regulations and alternatives in earnest. Continuous follow-up studies are required through a well-designed cohort.


Final results of the prospective European trial of the Endurant stent graft for endovascular abdominal aortic aneurysm repair.
The Endurant Stent Graft System (Medtronic Vascular, Santa Rosa, CA) is specifically designed to treat patients with abdominal aortic aneurysm, including those with difficult anatomies. This is the 1-year report of a prospective, non-randomised, open-label trial at 10 European centres. Between November 2007 and August 2008, 80 patients were enrolled for elective endovascular aneurysm repair (EVAR) with the Endurant; 71 with moderate (≤ 60°) and nine with high (60-75°) infrarenal aortic neck angulation. Safety and stent-graft performance were assessed throughout a 1-year follow-up period. The device was successfully delivered and deployed in all cases. All-cause mortality was 5% (4/80), with one possibly device-related death. Serious adverse events were comparable between the high and moderate angulation groups. There were no device migrations, stent fractures, aortic ruptures or conversions to open repair. Maximal aneurysm diameter decreased >5 mm in 42.7% of cases. A total of 28 endoleaks were observed (26 type II, two undetermined). Three secondary endovascular procedures were performed for outflow vessel stenosis, graft limb occlusion and iliac extension, resulting in a secondary patency rate of 100%. No re-interventions were required in the high angulation group. The Endurant Stent Graft was successfully delivered and deployed in all cases and performed safely and effectively in all patients, including those with unfavourable proximal neck anatomy.


Influence of osmolarity on lipopolysaccharides and virulence of Aeromonas hydrophila serotype O:34 strains grown at 37 degrees C.
Growth of Aeromonas hydrophila serotype O:34 strains at 37 degrees C at low and high osmolarity resulted in changes in the lipopolysaccharide (LPS) and virulence of the strains tested. We previously described the effect of growth temperature on LPS and virulence of these strains (S. Merino et al., Infect. Immun. 60:4343-4349, 1992). The effect of osmolarity can be observed when the cells grow at 37 degrees C but not when they grow at 20 degrees C. Purified LPS from cells cultivated at 37 degrees C and high osmolarity was smooth, while the LPS extracted from the cells cultivated at low osmolarity was rough. Furthermore, the strains were more virulent for fish and mice when they were grown at high osmolarity than when they were grown at low osmolarity and also showed increased extracellular activities when they were grown at high osmolarity. Finally, cells grown at high osmolarity showed better adhesion to HEp-2 cells than the same cells grown at low osmolarity, and furthermore the cells grown at high osmolarity were resistant to the bactericidal activity of nonimmune serum, while the same cells grown at low osmolarity were sensitive.


LncRNA UCA1 Participates in De Novo Synthesis of Guanine Nucleotides in Bladder Cancer by Recruiting TWIST1 to Increase IMPDH1/2.
Metabolic dysregulation has been identified as one of the hallmarks of cancer biology. Based on metabolic heterogeneity between bladder cancer tissues and adjacent tissues, we discovered several potential driving factors for the bladder cancer occurrence and development. Metabolic genomics showed purine metabolism pathway was mainly accumulated in bladder cancer. Long noncoding RNA urothelial carcinoma-associated 1 (LncRNA UCA1) is a potential tumor biomarker for bladder cancer diagnosis and prognosis, and it increases bladder cancer cell proliferation, migration, and invasion via the glycolysis pathway. However, whether UCA1 plays a role in purine metabolism in bladder cancer is unknown. Our findings showed that UCA1 could increase the transcription activity of guanine nucleotide de novo synthesis rate limiting enzyme inosine monophosphate dehydrogenase 1 (IMPDH1) and inosine monophosphate dehydrogenase 2 (IMPDH2), triggering in guanine nucleotide metabolic reprogramming. This process was achieved by UCA1 recruiting the transcription factor TWIST1 which binds to the IMPDH1and IMPDH2 promoter region. Increased guanine nucleotide synthesis pathway products stimulate RNA polymerase-dependent production of pre-ribosomal RNA and GTPase activity in bladder cancer cells, hence increasing bladder cancer cell proliferation, migration, and invasion. We have demonstrated that UCA1 regulates IMPDH1/2-mediated guanine nucleotide production via TWIST1, providing additional evidence of metabolic reprogramming.


Identification of Toxoplasma gondii cAMP dependent protein kinase and its role in the tachyzoite growth.
cAMP-dependent protein kinase (PKA) has been implicated in the asexual stage of the Toxoplasma gondii life cycle through assaying the effect of a PKA-specific inhibitor on its growth rate. Since inhibition of the host cell PKA cannot be ruled out, a more precise evaluation of the role of PKA, as well as characterization of the kinase itself, is necessary. The inhibitory effects of two PKA inhibitors, H89, an ATP-competitive chemical inhibitor, and PKI, a substrate-competitive mammalian natural peptide inhibitor, were estimated. In the in vitro kinase assay, the inhibitory effect of PKI on a recombinant T. gondii PKA catalytic subunit (TgPKA-C) was weaker compared to that on mammalian PKA-C. In a tachyzoite growth assay, PKI had little effect on the growth of tachyzoites, whereas H89 strongly inhibited it. Moreover, T. gondii PKA regulatory subunit (TgPKA-R)-overexpressing tachyzoites showed a significant growth defect. Our data suggest that PKA plays an important role in the growth of tachyzoites, and the inhibitory effect of substrate-competitive inhibitor PKI on T. gondii PKA was low compared to that of the ATP competitive inhibitor H89.


[Hypoxia stimulates the expression of HSP70 in pulmonary artery smooth muscle cells].
Hypoxic pulmonary hypertension is characterized by remodeling of pulmonary artery, proliferation and migration of pulmonary artery smooth muscle cells (PASMC), the mechanism of which was studied by observing the effect of hypoxia on DNA synthesis, cell cycle phase and transcription of heat shock protein 70 (HSP70). Our results indicate that hypoxia increases DNA synthesis, enhances the mitogenic effect of ET-1 on PASMC, stimulates quiescent PASMC entering S phase from G0/G1 phase and up-regulates the mRNA level of HSP70. Heat shock proteins are conservative and important stress proteins rendering the cells more tolerable to dangerous stresses by promoting growth and survival of the cells. All these findings suggest that heat shock proteins might also be important to the proliferation of PASMC induced by hypoxia and relevant to the study of the mechanism of hypoxic vascular remodeling.


Clinical Practice Guidelines for Bladder Cancer 2019 update by the Japanese Urological Association: Summary of the revision.
Despite just a 4-year interval from the last version (2015) of the Clinical Practice Guidelines for Bladder Cancer, several dramatic paradigm shifts have occurred in the latest clinical practice regarding both the diagnosis and treatment of bladder cancer. Herein, we updated the 2019 version of the Clinical Practice Guidelines for Bladder Cancer under the instruction of the Japanese Urological Association. We previously reported in a revision working position paper for Clinical Practice Guidelines for Bladder Cancer 2019 edition and described the methods of revision detail. The major points of change in the 2019 version are presented and explanations are given as follows: (i) introduction of the new reference assessment system; (ii) modification of the risk classification for non-muscle-invasive bladder cancer; (iii) addition of clinical questions for the new tumor-visible techniques in non-muscle-invasive bladder cancer; (iv) inclusion of minimally invasive surgeries for muscle-invasive bladder cancer and immune checkpoint inhibitors for locally advanced/metastatic muscle-invasive bladder cancer; (v) overview chapter of the histological variant of urothelial cancer and rare cancers of the bladder; and (vi) recommendation of follow up in non-muscle-invasive bladder cancer and muscle-invasive bladder cancer. Guidelines should be updated based on the current evidence and updates carried out without delay. The hope is that this guidelines will be assessed by many urologists and will be the cornerstone for the next revision.


The influence of the ACE inhibitor lisinopril on the glomerular metabolism of proteolytic enzymes in diabetic rats.
Clinical studies indicate a nephro-protective effect in conjunction with the use of ACE inhibitors. This study's aim was to determine whether ACE inhibitors influence the metabolism of glomerular cells in addition to their known hemodynamic effects. Streptozotocin diabetic rats were treated with lisinopril (DLis 1.5 mg/l water), or hydralazine (Dhyd, 50 mg/l water) over 4 weeks. Untreated diabetic rats (DC) and non-diabetic rats (C) served as controls. After four weeks of treatment, urinary excretion of albumin, blood pressure and metabolic control (Glyc-Hb) were measured. After treatment glomeruli were isolated and homogenized, and beta-NAG and total proteolytic activity against azocasein were measured. Glycated hemoglobin levels were similar in all diabetic groups (DC, 12%, Dhyd, 10%; DLis 11%). Blood pressure of DLis rats (79 +/- 3 mmHg) and DHyd rats (46 +/- 2 mmHg) was lower than that of DC rats (111 +/- 3 mmHg). Urinary albumin excretion of diabetic groups was lowest in DLis. Diabetic rats showed a decrease in glomerular beta-NAG (10 vs. 60.5 U/g protein) and total proteolytic activity against azocasein (148 vs. 170 U/mg protein hour) compared to non-diabetic rats. Lisinopril increased beta-NAG (30 vs. 14 U/g protein) and total proteolytic activity (160.5 vs. 141.5 U/mg protein hour) compared with hydralazine. Our study confirms that the nephro-protective effect of ACE inhibitors is partially due to modulatory effects on the metabolism of basement membrane proteins.


Effect of Primidone on Dentate Nucleus γ-Aminobutyric Acid Concentration in Patients With Essential Tremor.
It is not known whether current use of the medication primidone affects brain γ-aminobutyric acid (GABA) concentrations. This is an important potential confound in studies of the pathophysiology of essential tremor (ET), one of the most common neurological diseases. We compared GABA concentrations in the dentate nucleus in 6 ET patients taking primidone versus 26 ET patients not taking primidone. (1)H magnetic resonance spectroscopy was performed using a 3.0-T Siemens Tim Trio scanner. The MEGA-PRESS J-editing sequence was used for GABA detection in 2 cerebellar volumes of interest (left and right) that included the dentate nucleus. The right dentate GABA concentration was similar in the 2 groups (2.21 ± 0.46 [on primidone] vs 1.93 ± 0.39 [not on primidone], P = 0.15), as was the left dentate GABA concentration (1.61 ± 0.35 [on primidone] vs 1.67 ± 0.34 [not on primidone], P = 0.72). The daily primidone dose was not associated with either right or left dentate GABA concentrations (P = 0.89 and 0.76, respectively). We did not find a difference in dentate GABA concentrations between 6 ET patients taking daily primidone and 26 ET patients not taking primidone. Furthermore, there was no association between daily primidone dose and dentate GABA concentration. These data suggest that it is not necessary to exclude ET patients on primidone from magnetic resonance spectroscopy studies of dentate GABA concentration, and if assessment of these concentrations was to be developed as a biomarker for ET, primidone usage would not confound interpretation of the results.


Unexpected substituent effects in offset pi-pi stacked interactions in water.
We have measured the rotational barriers of meta- and para-substituted N-benzyl-2-(2-fluorophenyl)pyridinium bromides in aqueous solution by dynamic NMR as a model system for offset-stacking interactions in proteins. Because the benzyl ring can stack with the 2-fluorophenyl ring in the offset conformation in the ground state, but not the transition state, the rotational barrier reflects the magnitude of the stacking interaction. Only a small (0.1 kcal/mol) change in rotational barrier was found for para substituents relative to hydrogen. A much larger energy difference was found for electronegative meta substituents (up to 0.66 kcal/mol for CF3). Evidence suggests that this is due at least in part to an electrostatic interaction between electron-poor hydrogens on one ring with the electronegative substituents on the other ring.


[Rare differential diagnosis of a radicular spine syndrome: herpes zoster radiculitis].
We report on the case of a 66-year-old patient who was hospitalized because of intractable low back pain radiating into the right leg. Leg pain was accompanied by a numbness and muscle weakness which was clearly assigned to the L5 dermatome. Concerning the patient's medical history a nucleotomy L4/5 and a osteomyelofibrosis were known. MRI of the lumbar spine revealed a multisegmental stenosis which was pronounced on the level L4/5. One day after admission of the patient to the hospital a typical zoster exanthema involving the L5 dermatome appeared. Varicella-zoster virus (VZV) was detected in the fluid of the vesicular skin lesions by polymerase chain reaction. Intravenous administration of aciclovir lead to rapid decrease of pain and exanthema. A few months later the patient died because of an acute myeloid leukemia as a complication of the known osteomyelofibrosis. This case report shows that a herpes zoster infection can imitate a radicular spine syndrome usually caused by degenerative changes. Especially in immunocompromised patients, a zoster radiculitis should be included in the differential diagnosis of radiculopathy. VZV infection might also occur without skin lesions (zoster sine herpete) so that serological assays for the early detection of virus DNA can be useful.


Sudden cardiac death: directing the scope of resuscitation towards the heart and brain.
The fundamental goal of cardiopulmonary resuscitation (CPR) is recovery of the heart and the brain. This is best achieved by (1) immediate CPR for coronary and cerebral perfusion, (2) correction of the cause of cardiac arrest, and (3) controlled cardioplegic cardiac reperfusion. Failure of such an integrated therapy may cause permanent brain damage despite cardiac resuscitation. This strategy was applied at four centers to 34 sudden cardiac death patients (a) after acute myocardial infarction (n = 20), (b) "intraoperatively" following successful discontinuation of cardiopulmonary bypass (n = 4), and (c) "postoperatively" in the surgical ICU (n = 10). In each witnessed arrest the patient failed to respond to conventional CPR with ACLS interventions, including defibrillation. The cardiac arrest interval was 72 +/- 43 min (20-150 min). Compression and drugs maintained a BP > 60 mmHg to avoid cerebral hypoperfusion. Operating room (OR) transfer was delayed until the blood pressure was monitored. In four patients femoral bypass maintained perfusion while an angiographic diagnosis was made. Management principles included no repeat defibrillation attempts after 10 min of unsuccessful CPR, catheter-monitored peak BP > 60 mmHg during diagnosis and transit to the operating room, left ventricular venting during cardiopulmonary bypass and 20 min global and graft substrate enriched blood cardioplegic reperfusion. Survival was 79.4% with two neurological complications (5.8%). Recovery without adverse neurological outcomes is possible in a large number of cardiac arrest victims following prolonged manual CPR. Therapy is directed toward maintaining a monitored peak BP above 60 mmHg, determining the nature of the cardiac cause, and correcting it with controlled reperfusion to preserve function.


Multiplex Serum Biomarker Assays Improve Prediction of Renal and Mortality Outcomes in Chronic Kidney Disease.
We investigated the predictive value of 11 serum biomarkers for renal and mortality end points in people with CKD. Adults with CKD (n=139) were enrolled from outpatient clinics between February 2014 and November 2016. Biomarker quantification was performed using two multiplex arrays on a clinical-grade analyzer. Relationships between biomarkers and renal and mortality end points were investigated by random forests and Cox proportional hazards regression. The cohort was 56% male. The mean age was 63 years and median (IQR) CKD-EPI eGFR was 33 (24-51) ml/min per BSA. A total of 56 (40%) people developed a composite end point defined as ≥40% decline in eGFR, doubling of serum creatinine, RRT, or death over median (IQR) follow-up of 5.4 (4.7-5.7) years. Prediction of the composite end point was better with random forests trained on serum biomarkers compared with clinical variables (area under the curve of 0.81 versus 0.78). The predictive performance of biomarkers was further enhanced when considered alongside clinical variables (area under the curve of 0.83 versus 0.81 for biomarkers alone). Patients (n=27, 19%) with high soluble TNF receptor-1 (≥3 ng/ml) and neutrophil gelatinase-associated lipocalin (≥156 ng/ml), coupled with low complement 3a des-arginine (<2368 ng/ml), almost universally (96%) developed the composite renal and mortality end point. C-reactive protein (adjusted hazard ratio, 1.4; 95% CI, 1.1 to 1.8), neutrophil gelatinase-associated lipocalin (adjusted hazard ratio, 2.8; 95% CI, 1.3 to 6.1) and complement 3a desarginine (adjusted hazard ratio, 0.6; 95% CI, 0.4 to 0.96) independently predicted time to the composite end point. Outpatients with the triad of high soluble TNF receptor-1 and neutrophil gelatinase-associated lipocalin coupled with low complement 3a des-arginine had high adverse event rates over 5-year follow-up. Incorporation of serum biomarkers alongside clinical variables improved prediction of CKD progression and mortality. Our findings require confirmation in larger, more diverse patient cohorts.


[Effect of gum chewing on bowel function recovery in patients after radical cystectomy with urinary diversion].
To determine whether chewing gum during the postoperative period facilitates the recovery of bowel function in patients after radical cystectomy with ileum urinary diversion. In the study, 60 patients who underwent radical cystectomy followed by ileum urinary diversions during Nov. 2014 and Nov. 2015 in Department of Urology of Peking University First Hospital were randomized into three groups: gum chewing group, placebo group treated with the abdomen physical therapy machine and control group treated with ordinary method. Time to flatus, time to bowel movement, incidence of postoperative distension of the abdomen and abdominal pain, and gut related complications (such as ileus, intestinal fistula, and volrulus) of all the patients were recorded and analysed. In gum chewing group, the median time to flatus was 57 hours (49-72 hours), and the median time to bowel movement was 95 hours (88-109 hours), which were significantly shortened compared with the other two groups of patients (82 hours, 109 hours in placebo group and 81 hours, 108 hours in control group, respectively). No significant difference of the median time to flatus and to bowel movement was observed between placebo group and control group. There were no significant differences in the incidence of postoperative distension of the abdomen and abdominal pain, and gut related complications among the three groups. Chewing gum had stimulatory effect on bowel function recovery after cystectomy followed by ileum urinary diversion. Chewing gum was safe and simple, and could be routinely used for postoperative treatment after cystectomy and ileum urinary diversion.


[Dengue in Rio de Janeiro: rethinking popular participation in health].
This paper discusses popular participation in health from the point of view of an experience in dengue epidemic control in the late 1980s and early 1990s in the city of Rio de Janeiro. It refers specifically to a social movement called "Se Liga, Leopoldina", combining professionals (from health services and public schools) and members of popular organizations from the suburban neighborhoods served by the Leopoldina railway. The paper reviews the ideas motivating actions undertaken by the movement, principally the role of popular organizations in the control of endemics. It calls attention to the danger of preconceived notions when analyzing participation by members of popular organizations in the social movement. The author also calls attention to the fact that social scientists researching social movements should focus on being part of a wider social network of conviviality rather than seeing their role as educating members of the popular organization. Such networks should seek to strengthen and expand social support for the endemic control process.


In vitro-fermented raw and roasted walnuts induce expression of CAT and GSTT2 genes, growth inhibition, and apoptosis in LT97 colon adenoma cells.
Walnuts are rich in bioactive compounds such as polyunsaturated fatty acids, polyphenols, and dietary fiber. Therefore, the consumption of walnuts can contribute to a healthy diet and may reduce the risk for colon cancer. Heat treatment like roasting may change the chemical composition of walnuts and therefore their chemopreventive properties. Therefore, the hypothesis of the present study is that different roasting conditions (RCs) alter the chemopreventive effects of walnuts. Thus, the aim of the present study was to investigate whether different RCs (RC1=139.7°C/25 min, RC2=154.5°C/20 min, and RC3=185.5°C/25 min) alter the chemopreventive effects of walnuts. Raw and roasted walnuts were subjected to in vitro digestion and fermentation. After treatment of LT97 colon adenoma cells with fermentation supernatants (FSs), expression of CAT, SOD2, GPx1, GSTP1, and GSTT2 genes as well as cell growth and apoptosis was examined. In comparison to the fermentation blank control, walnut FS particularly increased mRNA levels of CAT 1.7-fold and GSTT2 3.1-fold, whereas GPx1 levels were significantly decreased 0.6-fold. Walnut FS decreased growth of adenoma cells in a time- and dose-dependent manner. In particular, higher concentrations of walnut FS (5%) significantly increased the number of early apoptotic cells 2.0-fold and induced caspase-3 activity 6.8-fold compared with the blank control. The roasting process had no direct impact on the observed effects. In sum, our results indicate that walnuts exhibit chemopreventive effects regarding the risk for colon cancer development by inducing expression of genes involved in detoxification (CAT, GSTT2) and by inducing growth inhibition and apoptosis in colon adenoma cells unaffected by moderate roasting.


Using support vector machines with multiple indices of diffusion for automated classification of mild cognitive impairment.
Few studies have looked at the potential of using diffusion tensor imaging (DTI) in conjunction with machine learning algorithms in order to automate the classification of healthy older subjects and subjects with mild cognitive impairment (MCI). Here we apply DTI to 40 healthy older subjects and 33 MCI subjects in order to derive values for multiple indices of diffusion within the white matter voxels of each subject. DTI measures were then used together with support vector machines (SVMs) to classify control and MCI subjects. Greater than 90% sensitivity and specificity was achieved using this method, demonstrating the potential of a joint DTI and SVM pipeline for fast, objective classification of healthy older and MCI subjects. Such tools may be useful for large scale drug trials in Alzheimer's disease where the early identification of subjects with MCI is critical.


A method for modeling noise in medical images.
We have developed a method to study the statistical properties of the noise found in various medical images. The method is specifically designed for types of noise with uncorrelated fluctuations. Such signal fluctuations generally originate in the physical processes of imaging rather than in the tissue textures. Various types of noise (e.g., photon, electronics, and quantization) often contribute to degrade medical images; the overall noise is generally assumed to be additive with a zero-mean, constant-variance Gaussian distribution. However, statistical analysis suggests that the noise variance could be better modeled by a nonlinear function of the image intensity depending on external parameters related to the image acquisition protocol. We present a method to extract the relationship between an image intensity and the noise variance and to evaluate the corresponding parameters. The method was applied successfully to magnetic resonance images with different acquisition sequences and to several types of X-ray images.


Tubulation of class II MHC compartments is microtubule dependent and involves multiple endolysosomal membrane proteins in primary dendritic cells.
Immature dendritic cells (DCs) capture exogenous Ags in the periphery for eventual processing in endolysosomes. Upon maturation by TLR agonists, DCs deliver peptide-loaded class II MHC molecules from these compartments to the cell surface via long tubular structures (endolysosomal tubules). The nature and rules that govern the movement of these DC compartments are unknown. In this study, we demonstrate that the tubules contain multiple proteins including the class II MHC molecules and LAMP1, a lysosomal resident protein, as well as CD63 and CD82, members of the tetraspanin family. Endolysosomal tubules can be stained with acidotropic dyes, indicating that they are extensions of lysosomes. However, the proper trafficking of class II MHC molecules themselves is not necessary for endolysosomal tubule formation. DCs lacking MyD88 can also form endolysosomal tubules, demonstrating that MyD88-dependent TLR activation is not necessary for the formation of this compartment. Endolysosomal tubules in DCs exhibit dynamic and saltatory movement, including bidirectional travel. Measured velocities are consistent with motor-based movement along microtubules. Indeed, nocodazole causes the collapse of endolysosomal tubules. In addition to its association with microtubules, endolysosomal tubules follow the plus ends of microtubules as visualized in primary DCs expressing end binding protein 1 (EB1)-enhanced GFP.


Accurate classification of plasma cell dyscrasias is achieved by combining artificial intelligence and flow cytometry.
Monoclonal gammopathy of unknown significance (MGUS), smouldering multiple myeloma (SMM), and multiple myeloma (MM) are very common neoplasms. However, it is often difficult to distinguish between these entities. In the present study, we aimed to classify the most powerful markers that could improve diagnosis by multiparametric flow cytometry (MFC). The present study included 348 patients based on two independent cohorts. We first assessed how representative the data were in the discovery cohort (123 MM, 97 MGUS) and then analysed their respective plasma cell (PC) phenotype in order to obtain a set of correlations with a hypersphere visualisation. Cluster of differentiation (CD)27 and CD38 were differentially expressed in MGUS and MM (P < 0·001). We found by a gradient boosting machine method that the percentage of abnormal PCs and the ratio PC/CD117 positive precursors were the most influential parameters at diagnosis to distinguish MGUS and MM. Finally, we designed a decisional algorithm allowing a predictive classification ≥95% when PC dyscrasias were suspected, without any misclassification between MGUS and SMM. We validated this algorithm in an independent cohort of PC dyscrasias (n = 87 MM, n = 41 MGUS). This artificial intelligence model is freely available online as a diagnostic tool application website for all MFC centers worldwide (https://aihematology.shinyapps.io/PCdyscrasiasToolDg/).


MiR-543-5p inhibits inflammation and promotes nerve regeneration through inactivation of the NF-κB in rats after spinal cord injury.
The aim is to determine the efficacy and specific mechanism of microRNA-543-5p (miR-543-5p) on spinal cord injury (SCI). The model of spinal cord injury was established in 6-week-old rats. Firstly, quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the changes of miR-543-5p in the spinal cord of rats in each group after spinal cord injury. Next, we observed the alterations in nuclear factor-kappa B (NF-κB) RNA level after injection of miR-543-5p. In addition, the enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of the inflammatory cytokines, and we used Western blotting to detect the protein associated with nerve regeneration at the protein levels. Finally, the Basso-Beattie-Bresnahan (BBB) rating scale was utilized to measure the recovery of hindlimbs function in rats. After spinal cord injury, the RNA expression of miR-543-5p in the rat spinal cord was decreased, and the RNA level of NF-κB was found to be decreased after the artificial injection of miR-543-5p. In the inflammatory expression, we found that the expression of various inflammatory mediators was also downregulated. However, the expression of nerve regeneration related factors was significantly upregulated, and it was observed that the evaluated score of the miR-543-5p group was higher than that of the SCI group within 1 month. MiR-543-5p inhibited NF-κB pathway and reduced the inflammatory factors, and ameliorated nerve regeneration, which ultimately promoted hindlimbs locomotor function.


Effective gastric acid suppression after oral administration of enteric-coated omeprazole granules.
Omeprazole is inactivated by exposure to gastric acid and is formulated as a gelatin capsule containing enteric-coated granules that release the drug in alkaline medium. In clinical situations where patients are unable to take the capsule orally, the optimum means of administration is uncertain. Eleven normal volunteers were given omeprazole 20 mg every day for one week before breakfast in random order as either a 20-mg capsule with water or free enteric-coated granules with either 8 oz of orange juice, 8 oz of water with 2 Alka-Seltzer antacid tablets (aspirin free), or 1 teaspoon of apple sauce. On day 7 of each regimen, an 8-hr intragastric pH study was performed following omeprazole 20 mg and standard breakfast. The median percentage of time of gastric acid pH > 4 after an omeprazole capsule was 68.5 (25-100); after granules with orange juice 59 (43-100); after granules in Alka-Seltzer solution 63 (31-100), and after granules in apple sauce 65 (30-99), with no significant differences (ANOVA). The time for the gastric pH to reach <4' after having been above was also similar for all four regimens (ANOVA). Omeprazole granules administered orally in a variety of ways achieve gastric acid suppression as effectively as the intact capsule.


Palliative surgery for recurrent bowel obstruction due to advanced ovarian cancer.
Intestinal obstruction is a frequent event in patients affected by ovarian carcinoma. Little data on repeat palliative surgery for recurrent bowel obstruction are available. The aim of this study was to analyze postoperative and long term outcomes of ovarian cancer patients who underwent reoperation for recurrent intestinal obstruction. We retrospectively evaluated the records of these patients treated at our Department between 1992 and 2002. Nine women with a mean age of 56 years (range 37-72) were identified. All patients had undergone previous abdominal surgery for bowel obstruction from ovarian cancer. All patients underwent exploratory laparotomy. In 4 patients (Group A) because of advanced disease, only exploratory surgery was carried out. A surgical correction was achieved in the other 5 patients (Group B), but only 3 patients had a successful palliation, defined as the ability to tolerate an oral intake for at least 60 days postoperatively. Postoperative mortality was nil, morbidity was 44.4%; particularly 2 patients developed an enterocutaneous fistula. Mean survival of Group A and B patients were 36.7 and 96.2 days respectively. The 3 successful palliated patients died of disease after 3.5, 4 and 5 months, in 2 cases for recurrent bowel obstruction. Repeat surgery for recurrent bowel obstruction in advanced ovarian carcinoma may achieve successful palliation in few cases and is associated with high postoperative morbidity and limited survival. In these patients non surgical approaches based on medical treatment, percutaneous endoscopic gastrostomy and stent placement should be considered.