Decreased insulin binding to adipocytes and circulating monocytes from obese subjects.
Insulin binding to isolated adipocytes from 16 normal and 14 obese patients was studied. The data indicated that, as a group, adipocytes from the obese patients bound significantly less insulin than normal. However, of the 14 obese patients, 5 were not hyperinsulinemic and 4 of these 5 subjects had normal insulin binding. These subjects were also younger, and had the onset of obesity in childhood. When these five patients were separated from the original 14 obese patients, enhanced differences in insulin binding to adipocytes were observed when normals and the remaining 9 obese subjects were compared. Similar findings were obtained with isolated circulating mononuclear cells from these same patients. Presumably the five normoinsulinemic obese patients were not insulin-resistant, and, thus, the data indicate that insulin binding to adipocytes was decreased only in insulin-resistant obese patients. This conclusion was strengthened by finding a highly significant correlation (r=-0.71, p less than 0.001) between insulin binding to adipocytes and fasting plasma insulin level, while a weaker correlation (r=-0.49,p less than 0.01) existed between insulin binding and degree of obesity. Finally, when insulin binding to adipocytes and mononuclear cells from the same individual was compared, a significant positive correlation was found (r=0.53,p less than 0.01). (a) insulin binding to adipocytes and mononuclear cells is decreased in cells from insulin-resistant obese patients; (b) a significant inverse relationship exists between fasting plasma insulin level and insulin binding to adipocytes; and (c) in obesity, events that affect insulin receptors on adipocytes similarly affect insulin receptors on mononuclear cells.


Short term effects of cigarette smoking on hospitalisation and associated lost workdays in a young healthy population.
There are relatively few published studies conducted among people of younger ages examining short term outcomes of cigarette smoking, and only a small number with outcomes important to employers. The present study was designed to assess the short term effects of smoking on hospitalisation and lost workdays. Retrospective cohort study. Military population. 87 991 men and women serving on active duty in the US Army during 1987 to 1998 who took a health risk appraisal two or more times and were followed for an average of 2.4 years. Rate ratios for hospitalisations and lost workdays, and fraction of hospitalisations and lost workdays attributable to current smoking (population attributable fraction). Compared with never smokers, men and women who were current smokers had higher short term rates of hospitalisation and lost workdays for a broad range of conditions. Population attributable fractions (PAFs) for outcomes not related to injury or pregnancy were 7.5% (men) and 5.0% (women) for hospitalisation, and 14.1% (men) and 3.0% (women) for lost workdays. Evidence suggests that current smoking may have been under reported in this cohort, in which case the true PAFs would be higher than those reported. In this young healthy population, substantial fractions of hospitalisations and lost workdays were attributable to current smoking, particularly among men.


Targeted Delivery of Proteasome Inhibitors to Somatostatin-Receptor-Expressing Cancer Cells by Octreotide Conjugation.
Clinical application of proteasome inhibitors (PIs) is so far limited to peripheral blood cancers due to the pronounced cytotoxicity towards all cell types. Targeted delivery of PIs could permit the treatment of other cancers along with decreasing side effects. Herein we describe the first small-molecule proteasome inhibitor conjugate for targeted delivery, created by fusing PIs to a synthetic ligand of somatostatin receptors, which are highly expressed in a variety of tumors. X-ray crystallographic studies and in vitro IC50 measurements demonstrated that addition of the cyclopeptide octreotide as a targeting vehicle does not affect the PI's binding mode. The cytotoxicity of the conjugate against somatostatin-receptor-expressing cells was up to 11-fold higher than that of a non-targeting surrogate. We have therefore established PIs as a new payload for drug conjugates and have shown that targeted delivery thereof could be a promising approach for the broader application of this FDA-approved class of compounds.


Global prevalence of traumatic non-fatal limb amputation.
Reliable information on both global need for prosthetic services and the current prosthetist workforce is limited. Global burden of disease estimates can provide valuable insight into amputation prevalence due to traumatic causes and global prosthetists needed to treat traumatic amputations. This study was conducted to quantify and interpret patterns in global distribution and prevalence of traumatic limb amputation by cause, region, and age within the context of prosthetic rehabilitation, prosthetist need, and prosthetist education. A secondary database descriptive study. Amputation prevalence and prevalence rate per 100,000 due to trauma were estimated using the 2017 global burden of disease results. Global burden of disease estimation utilizes a Bayesian metaregression and best available data to estimate the prevalence of diseases and injuries, such as amputation. In 2017, 57.7 million people were living with limb amputation due to traumatic causes worldwide. Leading traumatic causes of limb amputation were falls (36.2%), road injuries (15.7%), other transportation injuries (11.2%), and mechanical forces (10.4%). The highest number of prevalent traumatic amputations was in East Asia and South Asia followed by Western Europe, North Africa, and the Middle East, high-income North America and Eastern Europe. Based on these prevalence estimates, approximately 75,850 prosthetists are needed globally to treat people with traumatic amputations. Amputation prevalence estimates and patterns can inform prosthetic service provision, education and planning.


Effect of a stroke-specific follow-up care model on the quality of life of stroke patients and caregivers: A controlled trial.
To evaluate the effectiveness of a stroke-specific follow-up care model on quality of life for stroke patients, being discharged home, and their caregivers. A non-randomized, controlled trial, comparing an intervention group with a control group (usual care). Stroke patients and their caregivers. Intervention involved 5 home visits by a stroke care coordinator over a period of 18 months, using a structured assessment tool. Outcome measures were conducted at baseline (T0) and every 6 months thereafter (T6, T12 and T18) in the domains of quality of life (primary), activities of daily living, social activities, depression, anxiety and caregiver strain. The intervention group (n = 62) had significantly increased its social activities after 18 months, whereas the control group (n = 55) showed significantly decreased levels of social activities. In the first 6 months, levels of depression decreased significantly in caregivers of the intervention group. No differences were found for quality of life and the other outcome measures. The intervention was not effective in improving quality of life, but was effective in improving levels of social activities. The intervention may have focussed too much on screening for stroke-related problems and not as much on adequate follow-up care and referral.


Tunable Mixed Ionic/Electronic Conductivity and Permittivity of Graphene Oxide Paper for Electrochemical Energy Conversion.
Graphene oxide (GO) is a two-dimensional graphitic carbon material functionalized with oxygen-containing surface functional groups. The material is of interest in energy conversion, sensing, chemical processing, gas barrier, and electronics applications. Multilayer GO paper has recently been applied as a new proton conducting membrane in low temperature fuel cells. However, a detailed understanding of the electrical/dielectric properties, including separation of the ionic vs electronic contributions under relevant operating conditions, has so far been lacking. Here, the electrical conductivity and dielectric permittivity of GO paper are investigated in situ from 30 to 120 °C, and from 0 to 100% relative humidity (RH) using impedance spectroscopy. These are related to the water content, measured by thermogravimetric analysis. With the aid of electron blocking measurements, GO is demonstrated to be a mixed electronic-protonic conductor, and the ion transference number is derived for the first time. For RH > 40%, conductivity is dominated by proton transport (with a maximum of 0.5 mS/cm at 90 °C and 100% RH). For RH < 40%, electronic conductivity dominates (with a maximum of 7.4 mS/cm at ∼80 °C and 0% RH). The relative permittivity of GO paper increases with decreasing humidity, from ∼10 at 100% RH to several 1000 at 10% RH. These results underline the potential of GO for application not only as a proton conducting electrolyte but also as a mixed conducting electrode material under appropriate conditions. Such materials are highly applicable in electrochemical energy conversion and storage devices such as fuel cells and electrolyzers.


Comparing posttraumatic stress disorder's symptom structure between deployed and nondeployed veterans.
We tested two empirically validated 4-factor models of posttraumatic stress disorder (PTSD) symptoms using the PTSD Checklist: King, Leskin, King, and Weathers' (1998) model including reexperiencing, avoidance, emotional numbing, and hyperarousal factors, and Simms, Watson, and Doebbeling's (2002) model including reexperiencing, avoidance, dysphoria, and hyperarousal. Our aim was to determine which fit better in two groups of military veterans: peacekeepers previously deployed to a war zone (deployed group) and those trained for peacekeeping operations who were not deployed (nondeployed group). We compared the groups using multigroup confirmatory factor analysis. Adequate model fit was demonstrated among the nondeployed group, with no significant difference between King et al.'s (1998) model (separating avoidance and numbing) and Simms et al.'s (2002) similar model involving a dysphoria factor. A better fitting factor structure consistent with Simms et al.'s (2002) model was found in the deployed group. Comprehensive measurement invariance testing demonstrated significant differences between the deployed and nondeployed groups on all structural parameters, except observed variable intercepts (thus indicating similarities only in PTSD item severity). These findings add to researchers' understanding of PTSD's factor structure, given the revision of PTSD that will appear in the forthcoming 5th edition of the Diagnostic and Statistical Manual of Mental Disorders (American Psychiatric Association, 2010)--namely, that the factor structure may be quite different between groups with and without exposure to major traumatic events.


Energy balance feasibility study for Latinas in Texas: a qualitative assessment.
Obesity has reached epidemic levels, with nearly two-thirds of the U.S. population considered overweight or obese. Latinos have some of the highest rates of overweight, obesity, and sedentary lifestyle. Research from scientifically sound evidence-based interventions to reduce the disproportionate burden of obesity and its associated morbidity and mortality among Latinas is greatly needed. The purpose of this study was to assess knowledge, attitudes, and behaviors about nutrition and exercise among Latinas aged 40 years and older residing in a low-income community in Houston, Texas, and the applicability of an evidence-based church program to promote healthy energy balance. Qualitative assessment was conducted through 10 focus groups with 75 women recruited through three Catholic churches, community groups, and leaders. Participants identified barriers and enabling factors to healthy nutrition and physical activity. Barriers included lack of awareness about nutrition and physical activity, cultural beliefs, and socioeconomic and environmental factors. Preferred strategies were group activities with direct guidance from qualified individuals and interpersonal contact among participants, social support with positive reinforcement for behavior change or maintenance, and a friendly environment for learning and achieving suitable goals. The church was considered a powerful resource to influence Latinas to improve their health, exercise, and nutrition practices. Our findings suggest that using the church environment to reach Latina women aged 40 years and older is a feasible and culturally appropriate strategy. The church environment provides a safe, comfortable, and familiar atmosphere for women and addresses specific cultural barriers and safety concerns of family members.


What is the best way to diagnose and stage malignant pleural mesothelioma?
A best evidence topic in thoracic surgery was written according to a structured protocol. The question addressed was which diagnostic modality [computed tomography (CT), positron emission tomography (PET), combination PET/CT and magnetic resonance imaging (MRI)] provides the best diagnostic and staging information in patients with malignant pleural mesothelioma (MPM). Overall, 61 papers were found using the reported search, of which 14 represented the best evidence to answer the clinical question. The authors, journal, date and country of publication, patient group studied, study type, relevant outcomes and results are tabulated. We conclude that fluorodeoxyglucose (FDG)-PET is superior to MRI and CT but inferior to PET-CT, in terms of diagnostic specificity, sensitivity and staging of MPM. Four studies reported outcomes using FDG-PET to diagnose MPM. PET diagnosed MPM with high sensitivity (92%) and specificity (87.9%). Mean standardised uptake value (SUV) was higher in malignant than benign disease (4.91 vs. 1.41, P<0.0001). Lymph node metastases were detected with higher accuracy (80% vs. 66.7%) compared to extrathoracic disease. Three studies assessed the utility of PET-CT to diagnose MPM. Mean SUV was higher in malignant than benign disease (6.5 vs. 0.8, P<0.001). MPM was diagnosed with high sensitivity (88.2%), specificity (92.9%) and accuracy (88.9%). PET-CT had low sensitivity for stage N2 (38%) and T4 (67%) disease. CT-guided needle biopsy definitively diagnosed MPM after just one biopsy (100% vs. 9%) much more often than a 'blind' approach. CT had a lower success rate (92% vs. 100%) than thoracoscopic pleural biopsy but was equivalent to MRI in terms of detection of lymph node metastases (P=0.85) and visceral pleural tumour (P=0.64). CT had a lower specificity for stage II (77% vs. 100%, P<0.01) and stage III (75% vs. 100%, P<0.01) disease compared to PET-CT. Overall, the high specificity and sensitivity rates seen with open pleural biopsy make it a superior diagnostic modality to CT, MRI or PET for diagnosing patients with MPM.


Femtosecond-pulsed plasmonic nanotweezers.
We demonstrate for the first time plasmonic nanotweezers based on Au bowtie nanoantenna arrays (BNAs) that utilize a femtosecond-pulsed input source to enhance trapping of both Rayleigh and Mie particles. Using ultra-low input power densities, we demonstrate that the high-peak powers associated with a femtosecond source augment the trap stiffness to 2x that of nanotweezers employing a continuous-wave source, and 5x that of conventional tweezers using a femtosecond source. We show that for trapped fluorescent microparticles the two-photon response is enhanced by 2x in comparison to the response without nanoantennas. We also demonstrate tweezing of 80-nm diameter Ag nanoparticles, and observe an enhancement of the second-harmonic signal of ~3.5x for the combined nanoparticle-BNA system compared to the bare BNAs. Finally, under select illumination conditions, fusing of Ag nanoparticles to the BNAs is observed which holds potential for in situ fabrication of three-dimensional, bimetallic nanoantennas.


[The influence of cefteram pivoxil on the intestinal bacterial flora].
The influence of cefteram pivoxil (CFTM-PI), a new oral cephalosporin, on the intestinal bacterial flora was studied in tetra-contaminated mice and in pediatric patients. CFTM-PI in fine granular form was administered at a dose of 10 mg/kg once a day for 5 consecutive days to mice contaminated with 4 different species of organisms: Escherichia coli, Enterococcus faecalis, Bacteroides fragilis and Bifidobacterium breve. No remarkable change was observed in the fecal viable cell count except that slight decrease in E. coli count was observed on days 2 to 5 after starting administration. The subjects in pediatric study were 5 children, 3 boys and 2 girls at ages from 6 months to 10 years 4 months, with infections. Their body weights ranged from 3.5 to 28.0 kg. CFTM-PI in fine granular form was administered at each dose of 3.0 to 3.8 mg/kg, 3 times daily for 5 to 11 days. During the administration of CFTM-PI, there were some variations in the change of the fecal bacterial flora noticed between subjects. Although Enterobacteriaceae tended to decrease and Enterococci tended to increase, other main aerobes and anaerobes were almost unchanged in most cases. There was no case in which glucose non-fermenting Gram-negative rods and fungi became predominant. In a 10 years 4 months old boy, the fecal concentration of CFTM, the active form of CFTM-PI, was 72.20 micrograms/g and a remarkable decrease of total anaerobe count was observed. In feces, CFTM-PI and CFTM were detected in 4 and 2 cases respectively, and their concentrations were 1.50 approximately 89.65 micrograms/g and 2.25 approximately 72.20 micrograms/g, respectively. beta-Lactamase activities in feces were positive in all cases. From the above, CFTM-PI is considered to be a drug with relatively less influence on the intestinal bacterial flora. But as high concentrations of drugs were detected in feces under some circumstances, we need to pay attention to fecal drug concentrations.


A novel non-thermal non-ablative full panel LED photomodulation device for reversal of photoaging: digital microscopic and clinical results in various skin types.
Photomodulation is a process that manipulates or regulates cell activity using light sources without thermal effect. Previous studies of LED photomodulation have shown skin textural improvement accompanied by increased collagen deposition with reduced MMP-1 (collagenase) activity in the papillary dermis. The purpose of this study was to investigate a separate cohort of patients (N =93) with a wide range of Fitzpatrick skin types treated by LED photomodulation using the Gentlewaves full panel 590 nm high energy LED array with a specific sequence or code of pulsing in the millisecond domain. Results showed improvement of signs of photoaging in 90%. The majority of patients demonstrated improvement in peri-ocular wrinkles, reduction in Fitzpatrick photoaging classification, global skin texture and background erythema, and pigmentation. No side effects were noted. LED photomodulation is a safe and effective non-painful non-ablative modality for improvement of photoaging.


Effect of dietary raw and fermented sour cherry kernel (Prunus cerasus L.) on digestibility, intestinal morphology and caecal microflora in broiler chickens.
This study was conducted to investigate the effect of dietary raw sour cherry kernel (RC) or fermented sour cherry kernel (FC) on apparent digestibility, ileal morphology, and caecal microflora in broiler chickens. Raw sour cherry kernel was fermented by Aspergillus niger for 7 D. A total of 343 one-day-old Ross 308 male chicks were assigned to 7 dietary treatments consisting of 7 replicates of 7 broilers each. All birds were fed with a commercial diet or diets supplemented with 1%, 2%, or 4% RC or FC. The experimental period was 42 D. Apparent dry matter (DM), nitrogen and ash digestibilities were diminished (P < 0.05) by dietary RC inclusion, although dietary FC did not negatively affect (P > 0.05) nutrient digestibility. Dietary 1% FC increased (P < 0.01) the villus height to crypt depth ratio (VH:CD) compared with the other treatment groups, although RC4 reduced the villus height (VH, P < 0.001) and VH:CD (P < 0.01), compared with the control group. Dietary treatments had no effect (P > 0.05) on the crypt depth (CD). Birds fed 1% FC had the highest (P < 0.05) caecal Lactobacillus spp. counts among the treatment groups. Enterococcus spp. and Escherichia coli counts in cecum were not affected (P > 0.05) by dietary treatments. The results showed that the dietary inclusion of 1% FC improved ileal morphology and caecal microflora without any adverse effect on the apparent digestibility. These results indicate that FC has the potential to be a feed additive which improves intestinal health for broiler diets.


Changes in coronary heart disease risk profile of adults with intellectual disabilities following a physical activity intervention.
Regular physical activity is one of the modifiable risk factors for coronary heart disease (CHD). With an increasing age profile and similar patterns of morbidity to the general population, persons with intellectual disabilities (ID) and their caregivers would benefit from data that indicate CHD risk factors. Knowledge of the CHD risk factors and the changes a physical activity intervention may have on theses risk factors will facilitate future intervention programmes. A cohort of 100 men and women between the ages of 21 and 73 years with ID living in a community group home in the North-West Province of South Africa was recruited. A CHD risk profile was compiled by means of a questionnaire and physical assessment that included resting blood pressure, body mass index, non-fasting glucose and cholesterol and cardiorespiratory fitness. A 12-week physical activity intervention was then conducted 3 days/week after which the baseline measurements were repeated. The results indicated that 85% of the participants were inactive, while 67% were overweight and obese. Hypertension (6.1%) and smoking (6.1%) were relatively low in this population with ID. Glucose concentrations above the recommended cut-off values were observed in 28% of the participants. Total cholesterol concentrations above normal were measured in 23% of the participants. The physical activity intervention reduced inactivity to 50% and resulted in a significant increase in cardiorespiratory fitness and a decrease in percentage body fat in both men and women. Inactivity is a major risk factor in this population with ID living in a community group setting. The implementation of the physical activity intervention significantly reduced the risk factors for CHD.


In situ biomechanics of the syrinx and sound generation in pigeons.
The in situ biomechanics of the vocal organ, the syrinx, was studied in anesthetized pigeons using fiberoptic instruments. The role of syringeal muscles was determined by electrical stimulation, and phonation was induced by injecting gas into the subsyringeal air sacs. This study presents the first direct observations of the biomechanical processes that occur in an intact syrinx. Contraction of one of the syringeal muscles, the m. tracheolateralis (TL), withdraws the lateral tympaniform membranes (LTM) from the syringeal lumen, causing opening of the syringeal airways. Shortening of a second muscle, the sternotrachealis (ST), draws the syringeal cartilages closer to each other, causing the LTM to fold into the syringeal lumen. Maximal ST contraction does not lead to complete closure of the syrinx. As air-sac pressure is increased by the injection of gas, the LTM are drawn into the syringeal lumen and balloon in a rostral direction until they touch, thus forming a fold-like valve. Air-induced phonation is always associated with vibrations of the membrane folds, suggesting that pulsatile release of air into the trachea by vibratory motion of the LTM generates sound. During air-induced phonation, strong stimulation of the TL terminates sound generation by abducting the LTM, whereas weak stimulation changes the geometry of the membrane folds, which is accompanied by changes in the acoustic structure of the sound. Stimulation of the ST has little effect on air-induced sounds. The LTM appear to be the main sound generators, since disabling the medial tympaniform membranes (MTM) with tissue adhesive does not prevent phonation or change the frequency and amplitude structure of display coos in spontaneously vocalizing pigeons. Moreover, the activity of the syringeal muscles appears to have a mainly modulatory function, suggesting that the basic sound-generating mechanism is similar in both air-induced and natural phonation.


Sensitivity and predictive value of anti-GM1/galactocerebroside IgM antibodies in multifocal motor neuropathy.
Increased titres of serum IgM antibodies to GM1 ganglioside are often associated with multifocal motor neuropathy (MMN). Testing for IgM antibodies to other antigens including GM2, the mixture of GM1 and galactocerebroside (GM1/GalC) and the disulfated heparin disaccharide NS6S were reported to increase the sensitivity of antibody testing in MMN even if it is unclear whether the specificity and positive (PPV) or negative predictive value (NPV) for MMN were also affected. We measured IgM antibodies to GM1, GM2, galactocerebroside, GM1/GalC and NS6S in 40 consecutive patients with MMN and 142 controls with other neuropathies or related diseases and compared their sensitivity, specificity and PPV for MMN. With the only exception of anti-GM2 and, partially, anti-NS6S antibodies, IgM antibodies to the antigens tested were more frequent in MMN than in controls. Increased titres of anti-GM1 IgM were found in 48% of MMN patients with a specificity of 93% and PPV for MMN of 66%. Anti-GM1/GalC antibodies were present in all anti-GM1 positive MMN patients and in 11 additional patients (28%) with MMN raising the sensitivity of antibody testing to 75%. The specificity (85%) and PPV (59%) for MMN was, however, moderately reduced compared to anti-GM1 IgM, even if they rose with increasing anti-GM1/GalC titres. IgM antibodies to GM2, NS6S and galactocerebroside were found in 8%, 23% and 60% of MMN patients but had a low specificity and PPV for MMN. Testing for anti-GM1/GalC IgM significantly increased the sensitivity of antibody testing in MMN compared to anti-GM1 alone (p=0.021) and may represent a preferred option for GM1 reactivity testing in MMN.


Contribution of a Blood-Based Protein Biomarker Panel to the Classification of Indeterminate Pulmonary Nodules.
The workup and longitudinal monitoring for subjects presenting with pulmonary nodules is a pressing clinical problem. A blood-based biomarker panel potentially has utility for identifying subjects at higher risk for harboring a malignant nodule for whom additional workup would be indicated or subjects at reduced risk for whom imaging-based follow-up would be indicated. To assess whether a previously described four-protein biomarker panel, reported to improve assessment of lung cancer risk compared with a smoking-based lung cancer risk model, can provide discrimination between benign and malignant indeterminate pulmonary nodules. A previously validated multiplex enzyme-linked immunoassay was performed on matched case and control samples from each cohort. The biomarker panel was tested in two case-control cohorts of patients presenting with indeterminate pulmonary nodules at the University of Pittsburgh Medical Center and the University of Texas Southwestern. In both cohorts, the biomarker panel resulted in improved prediction of lung cancer risk over a model on the basis of nodule size alone. Of particular note, the addition of the marker panel to nodule size greatly improved sensitivity at a high specificity in both cohorts. A four-marker biomarker panel, previously validated to improve lung cancer risk prediction, was found to also have utility in distinguishing benign from malignant indeterminate pulmonary nodules. Its performance in improving sensitivity at a high specificity indicates potential utility of the marker panel in assessing likelihood of malignancy in otherwise indeterminate nodules.


Phage-induced diversification improves host evolvability.
Bacteriophage (viruses that infect bacteria) are of key importance in ecological processes at scales from biofilms to biogeochemical cycles. Close interaction can lead to antagonistic coevolution of phage and their hosts. Selection pressures imposed by phage are often frequency-dependent, such that rare phenotypes are favoured; this occurs when infection depends on some form of genetic matching. Also, resistance to phage often affects host fitness by pleiotropy (whereby mutations conferring resistance affect the function of other traits) and/or direct costs of resistance mechanisms. Here a simple model of bacteria and bacteriophage coevolving in a resource-limited chemostat is used to study the effect of coevolving phage on the evolution of bacterial hosts. Density-dependent mortality from phage predation limits the density of any single bacterial strain, preventing competitive exclusion by faster-growing strains. Thus multiple strains can coexist by partitioning resources and stable high diversity is created by negative frequency-dependent selection from phage. Standing bacterial diversity promotes adaptation in dynamic environments, since it increases the likelihood of a pre-existing genotype being suited to altered conditions. In addition, frequency-dependent selection for resistance creates transient local trade-offs between growth rate and resistance that allow bacterial strains to adapt across fitness valleys. Thus bacterial populations that (in the absence of phage) would have been trapped at sub-optimal local peaks in the adaptive landscape are able (in the presence of phage) to reach alternate higher peaks than could have been reached by mutation alone. This study shows that reasonable assumptions for coevolution of bacteria and phage create conditions in which phage increase the evolutionary potential of their hosts. Thus phage, in contrast to their deleterious effects on individual host cells, can confer an evolutionary benefit to bacterial populations. These findings have implications for the role of phage in ecosystem processes, where they have mainly been considered as a mortality factor; these results suggest that on long timescales phage may actually increase bacterial productivity by aiding the evolution of faster-growing strains. Furthermore, these results suggest that the therapeutic use of phage to treat bacterial infections (phage therapy) could have unintended negative side-effects.


Chemopreventive and therapeutic potentials of thymoquinone in HepG2 cells: mechanistic perspectives.
Liver cancer is the fifth commonest malignancy worldwide and the third leading cause of death. Identifying novel curative and preventive therapy may improve its prognosis. In this study, thymoquinone (TQ), the most active biological ingredient of Nigella sativa Linn, was investigated for its antitumor activity. Mechanistic perspectives underlying this antitumor activity were explored by testing its effect on cell cycle, apoptosis, and angiogenesis. In addition, the chemopreventive effect of TQ was carried out by measuring its effect on phase I CYP1A1 and phase II glutathione S-transferase (GST) drug-metabolizing enzymes. The results of the present study revealed the effectiveness of TQ as an antitumor agent against different types of cancer including brain, colon, cervix and liver at both a time- and concentration-dependent manner. In HepG2 cells, it induced G2/M phase cell cycle arrest and a concentration-dependent increase in the percentage of apoptotic cells with an increase in the ratio of Bax/BCL-2. Moreover, the expression of mRNA and protein level of vascular endothelial growth factor decreased as the concentration of TQ increased. Our data showed a significant inhibition of induced phase I CYP1A1 enzyme, and elevation in the content of glutathione and activity of phase II enzyme GST, in HepG2 cells. Our results provide support for the beneficial use of TQ as a therapeutic and chemopreventive agent against liver cancer.


Prediction of protein-protein interactions in dengue virus coat proteins guided by low resolution cryoEM structures.
Dengue virus along with the other members of the flaviviridae family has reemerged as deadly human pathogens. Understanding the mechanistic details of these infections can be highly rewarding in developing effective antivirals. During maturation of the virus inside the host cell, the coat proteins E and M undergo conformational changes, altering the morphology of the viral coat. However, due to low resolution nature of the available 3-D structures of viral assemblies, the atomic details of these changes are still elusive. In the present analysis, starting from Calpha positions of low resolution cryo electron microscopic structures the residue level details of protein-protein interaction interfaces of dengue virus coat proteins have been predicted. By comparing the preexisting structures of virus in different phases of life cycle, the changes taking place in these predicted protein-protein interaction interfaces were followed as a function of maturation process of the virus. Besides changing the current notion about the presence of only homodimers in the mature viral coat, the present analysis indicated presence of a proline-rich motif at the protein-protein interaction interface of the coat protein. Investigating the conservation status of these seemingly functionally crucial residues across other members of flaviviridae family enabled dissecting common mechanisms used for infections by these viruses. Thus, using computational approach the present analysis has provided better insights into the preexisting low resolution structures of virus assemblies, the findings of which can be made use of in designing effective antivirals against these deadly human pathogens.


Improved reliability for fractal dimension calculation of the vascular imprints on the cranial vault mapped with topographical correction.
We presented a novel approach to studies of the vascular grooves located on the inner surface of the cranial vault. A three-dimensional vision system that acquired the endocranial surface topography was used for this purpose. The acquired data were used to generate images showing the branching pattern of the middle meningeal artery. Fractal dimension was used to characterize and analyze branching pattern complexity. We discussed the usefulness of the latter method and indicated difficulties and potential errors connected to the fractal dimension application. The technique introduced for recording traits of the object surface appears to be helpful in anatomical study of morphological variation of dural vascularization. It may also be applicable in paleoneurological research based on analysis of the cranial remnants. Fractal dimension should be used carefully as a method sensitive to many aspects of data acquisition and processing.


Hormone therapy improves femur geometry among ethnically diverse postmenopausal participants in the Women's Health Initiative hormone intervention trials.
Loss of bone strength underlies osteoporotic fragility fractures. We hypothesized that hormone interventions significantly improve the structural geometry of proximal femur cross-sections. Study participants were from the Women's Health Initiative hormone intervention trials: either the conjugated equine estrogen (CEE) only (N(placebo) = 447, N(CEE) = 422) trial or the estrogen (E) plus progestin (P) (N(placebo) = 441, N(E+P) = 503) trial, who were 50-79 yr old at baseline and were followed up to 6 yr. BMD scans by DXA were conducted at baseline, year 1, year 3, and year 6. Femur geometry was derived from hip DXA scans using the hip structural analysis (HSA) method. Mixed effects models with the intent-to-treat analysis approach were used. There were no significant differences in treatment effects between the E-alone and the E + P trial, so the analyses were conducted with participants combined from both trials. Treatment benefits (p < 0.05) on femur geometry were observed as early as 1 yr after the intervention. From baseline to year 6, section modulus (a measure of maximum bending stress) was preserved, and buckling ratio (an index of cortical instability under compression) was reduced by hormone interventions (p < 0.05); the differences in the percent changes from baseline to year 6 between women on hormone intervention versus women on placebo were 2.3-3.6% for section modulus and -5.3% to - 4.3% for buckling ratio. Hormone interventions led to favorable changes in femur geometry, which may help explain the reduced fracture risk observed in hormone interventions.


Immature stages of Spodoptera albula (Walker) (Lepidoptera: Noctuidae): developmental parameters and host plants.
This study aimed to detail the temporal and morphological parameters of the immature stages of Spodoptera albula (Walker 1857) under controlled conditions (25 ± 1°C, 70 ± 10% RH and 14 hour photophase) and to gather information about their larval host plants. For this purpose, a new rearing method and artificial diet was employed and validated. The viability of the egg, larval, pupal and pre-pupal stages was 94.54, 97.33, 93.84 and 92.34%, respectively. The average duration of the egg, larval, pupal and pre-pupal stages was 4.14, 16.37, 1.69, and 9.34 days, respectively. During the larval stage, 80.85% of females and 93.99% of males passed through six and remaining through seven instars, with significant larval protandry. The larvae that developed through six and seven instars exhibited a mean growth rate of 1.58 and 1.48, respectively. Fifty five host plant species belonging to 29 families are listed. The female pupae were significantly larger, exhibiting protogyny. Both the rearing methods as well as the larval diet proved adequate, providing more detailled observations of the biological cycle, especially the larval stage, and resulting in an overall survival of almost 80%.


Isolates from Colonic Spirochetosis in Humans Show High Genomic Divergence and Potential Pathogenic Features but Are Not Detected Using Standard Primers for the Human Microbiota.
Colonic spirochetosis, diagnosed based on the striking appearance in histological sections, still has an obscure clinical relevance, and only a few bacterial isolates from this condition have been characterized to date. In a randomized, population-based study in Stockholm, Sweden, 745 healthy individuals underwent colonoscopy with biopsy sampling. Of these individuals, 17 (2.3%) had colonic spirochetosis, which was associated with eosinophilic infiltration and a 3-fold-increased risk for irritable bowel syndrome (IBS). We aimed to culture the bacteria and perform whole-genome sequencing of the isolates from this unique representative population sample. From 14 out of 17 individuals with spirochetosis we successfully isolated, cultured, and performed whole-genome sequencing of in total 17 isolates, including the Brachyspira aalborgi type strain, 513A. Also, 16S analysis of the mucosa-associated microbiota was performed in the cases and nonspirochetosis controls. We found one isolate to be of the species Brachyspira pilosicoli; all remaining isolates were of the species Brachyspira aalborgi Besides displaying extensive genetic heterogeneity, the isolates harbored several mucin-degrading enzymes and other virulence-associated genes that could confer a pathogenic potential in the human colon. We also showed that 16S amplicon sequencing using standard primers for human microbiota studies failed to detect Brachyspira due to primer incompatibility.IMPORTANCE This is the first report of whole-genome analysis of clinical isolates from individuals with colonic spirochetosis. This characterization provides new opportunities in understanding the physiology and potentials of these bacteria that densely colonize the gut in the individuals infected. The observation that standard 16S amplicon primers fail to detect colonic spirochetosis may have major implications for studies searching for associations between members of the microbiota and clinical conditions such as irritable bowel syndrome (IBS) and should be taken into consideration in project design and interpretation of gastrointestinal tract microbiota in population-based and clinical settings.


Adrenomedullin inhibits neurotransmission of calcitonin gene-related peptide (CGRP)-containing vasodilator nerves in rat mesenteric resistance arteries.
We have reported that the rat mesenteric resistance artery has innervation of calcitonin gene-related peptide (CGRP)-containing vasodilator nerves (CGRPergic nerves). We also demonstrated that adrenomedullin (AM) causes mesenteric vasodilation through activation of CGRP receptors. The present study was designed to examine the effect of AM on neurotransmission of CGRPergic nerves in rat mesenteric arteries. In preconstricted preparations without endothelium, periarterial nerve stimulation (PNS, 1 and 2 Hz) induced a frequency-dependent vasodilation. A bolus injection of CGRP (10 pmol) into the perfusate also caused a vasodilation. AM (0.1 to 10 nM) concentration-dependently caused 40% to 60% inhibition of the PNS-induced vasodilation, but AM did not attenuate vasodilation induced by exogenous CGRP injection. The inhibitory effect of AM (10 nM) on PNS-induced vasodilation was further potentiated by CGRP [8-37] (CGRP receptor antagonist, 50 nM), which attenuated the vasodilator response to the CGRP injection. Combined perfusion of AM [22-52] (AM receptor antagonist, 10 to 100 nM) resulted in further inhibition of PNS-induced neurogenic vasodilation without affecting the vasodilator response to the CGRP injection. CGRP [8-37] but not AM [22-52] antagonized vasodilation induced by AM perfusion. These findings suggest that AM presynaptically inhibits neurotransmission of CGRPergic nerves, probably decreasing CGRP release, via receptors different from CGRP receptors.


RNA templating the epigenome: long noncoding RNAs as molecular scaffolds.
Cellular pathways must be synergized, controlled and organized to manage homeostasis. To achieve high selectivity within the crowded cellular milieu the cell utilizes scaffolding complexes whose role is to bring molecules in proximity thereby controlling and enhancing intermolecular interactions and signaling events. To date, scaffolds have been shown to be composed of proteinaceous units; however, recent evidence has supported the idea that non-coding RNAs may also play a similar role. In this point of view article we discuss recent data on ncRNA scaffolds, with particular focus on ncRNA HOTAIR. Using our current knowledge of signaling networks we discuss the role that RNA may play in writing and regulating histone modifications and the information needed for correct gene expression. Further, we speculate on additional, yet undiscovered roles that ncRNAs may be playing as molecular scaffolds.


Electromagnetic susceptibility anisotropy and its importance for paramagnetic NMR and optical spectroscopy in lanthanide coordination chemistry.
The importance of the directional dependence of magnetic susceptibility in magnetic resonance and of electric susceptibility in the optical spectroscopy of lanthanide coordination complexes is assessed. A body of more reliable shift, relaxation and optical emission data is emerging for well-defined isostructural series of complexes, allowing detailed comparative analyses to be undertaken. Such work is highlighting the limitations of the current NMR shift and relaxation theories, as well as emphasising the absence of a compelling theoretical framework to explain optical emission phenomena.


Early trophic feeding versus enteral fasting for very preterm or very low birth weight infants.
The introduction of enteral feeds for very preterm (< 32 weeks) or very low birth weight (< 1500 grams) infants is often delayed due to concern that early introduction may not be tolerated and may increase the risk of necrotising enterocolitis. However, prolonged enteral fasting may diminish the functional adaptation of the immature gastrointestinal tract and extend the need for parenteral nutrition with its attendant infectious and metabolic risks. Trophic feeding, giving infants very small volumes of milk to promote intestinal maturation, may enhance feeding tolerance and decrease the time taken to reach full enteral feeding independently of parenteral nutrition. To determine the effect of early trophic feeding versus enteral fasting on feed tolerance, growth and development, and the incidence of neonatal morbidity (including necrotising enterocolitis and invasive infection) and mortality in very preterm or VLBW infants. We used the standard search strategy of the Cochrane Neonatal Review Group. This included electronic searches of the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2012, Issue 12), MEDLINE, EMBASE and CINAHL (1980 until December 2012), conference proceedings and previous reviews. Randomised or quasi-randomised controlled trials that assessed the effects of early trophic feeding (milk volumes up to 24 ml/kg/day introduced before 96 hours postnatal age and continued until at least one week after birth) versus a comparable period of enteral fasting in very preterm or very low birth weight infants. We extracted data using the standard methods of the Cochrane Neonatal Review Group with separate evaluation of trial quality and data extraction by two authors and synthesis of data using risk ratio, risk difference and mean difference. Nine trials in which a total of 754 very preterm or very low birth weight infants participated were eligible for inclusion. Few participants were extremely preterm (< 28 weeks) or extremely low birth weight (< 1000 grams) or growth restricted. These trials did not provide any evidence that early trophic feeding affected feed tolerance or growth rates. Meta-analysis did not detect a statistically significant effect on the incidence of necrotising enterocolitis: typical risk ratio 1.07 (95% confidence interval 0.67 to 1.70); risk difference 0.01 (-0.03 to 0.05). The available trial data do not provide evidence of important beneficial or harmful effects of early trophic feeding for very preterm or very low birth weight infants. The applicability of these findings to extremely preterm, extremely low birth weight or growth restricted infants is limited. Further randomised controlled trials would be needed to determine how trophic feeding compared with enteral fasting affects important outcomes in this population.


Varicella Viruses Inhibit Interferon-Stimulated JAK-STAT Signaling through Multiple Mechanisms.
Varicella zoster virus (VZV) causes chickenpox in humans and, subsequently, establishes latency in the sensory ganglia from where it reactivates to cause herpes zoster. Infection of rhesus macaques with simian varicella virus (SVV) recapitulates VZV pathogenesis in humans thus representing a suitable animal model for VZV infection. While the type I interferon (IFN) response has been shown to affect VZV replication, the virus employs counter mechanisms to prevent the induction of anti-viral IFN stimulated genes (ISG). Here, we demonstrate that SVV inhibits type I IFN-activated signal transduction via the JAK-STAT pathway. SVV-infected rhesus fibroblasts were refractory to IFN stimulation displaying reduced protein levels of IRF9 and lacking STAT2 phosphorylation. Since previous work implicated involvement of the VZV immediate early gene product ORF63 in preventing ISG-induction we studied the role of SVV ORF63 in generating resistance to IFN treatment. Interestingly, SVV ORF63 did not affect STAT2 phosphorylation but caused IRF9 degradation in a proteasome-dependent manner, suggesting that SVV employs multiple mechanisms to counteract the effect of IFN. Control of SVV ORF63 protein levels via fusion to a dihydrofolate reductase (DHFR)-degradation domain additionally confirmed its requirement for viral replication. Our results also show a prominent reduction of IRF9 and inhibition of STAT2 phosphorylation in VZV-infected cells. In addition, cells expressing VZV ORF63 blocked IFN-stimulation and displayed reduced levels of the IRF9 protein. Taken together, our data suggest that varicella ORF63 prevents ISG-induction both directly via IRF9 degradation and indirectly via transcriptional control of viral proteins that interfere with STAT2 phosphorylation. SVV and VZV thus encode multiple viral gene products that tightly control IFN-induced anti-viral responses.


Correspondence in pictorial space.
We have investigated psychophysically determined image correspondences between pairs of photographs of a single three-dimensional (3-D) object in various poses. These correspondences were obtained by presenting the pictures simultaneously, side by side, and letting the subject match a marker in one picture with a marker (under manual control) in the other picture. Between poses, the object was rotated about a fixed vertical axis; thus, the shifts of the veridical correspondences (with respect to the surface of the object) were very nearly horizontal. In fact, the subjects produced appreciable scatter in both horizontal and vertical directions. The scatter in repeated sessions and between data depends on the local (landmarks) and global (interpolation) structure of the pictures. Since the object was fairly smooth (white semigloss finish) and nontextured, the only way to establish the correspondence is by way of the "pictorial relief." The relief is some largely unknown function of the image structure and the observer. Apparently, more immediate entities (e.g., the shading or the contour) cannot be used as such, since they vary with the pose. We compare these data with results obtained with a surface attitude probe on a single picture. We studied various measures of consistency both within a single method and between methods. We found that subjects were confident in establishing correspondences, but results scattered appreciably in a way that depended on both global and local image structure. Correspondence results for various pose angles were mutually very consistent, but only to a minor extent with results of attitude measurements. The main finding was that subjects could establish correspondence on the basis of their 3-D interpretation (pictorial relief), even if the 2-D graytone distributions are quite different.


Pressure Optimized PowEred Respirator (PROPER): A miniaturized wearable cleanroom and biosafety system for aerially transmitted viral infections such as COVID-19.
The supply of Personal Protective Equipment (PPE) in hospitals to keep the Health Care Professionals (HCP) safe taking care of patients may be limited, especially during the outbreak of a new disease. In particular, the face and body protective equipment is critical to prevent the wearer from exposure to pathogenic biological airborne particulates. This situation has been now observed worldwide during the onset of the COVID-19 pandemic. As concern over shortages of PPE at hospitals grows, we share with the public and makers' community the Pressure Optimized PowEred Respirator (PROPER) equipment, made out of COTS components. It is functionally equivalent to a Powered Air Purifying Respirator (PAPR). PROPER, a hood-based system which uses open source and easily accessible components is low-cost, relatively passive in terms of energy consumption and mechanisms, and easy and fast to 3D print, build and assemble. We have adapted our experience on building clean room environments and qualifying the bioburden of space instruments to this solution, which is in essence a miniaturized, personal, wearable cleanroom. PROPER would be able to offer better protection than an N95 respirator mask, mainly because it is insensitive to seal fit and it shields the eyes as well. The PROPER SMS fabric is designed for single-use and not intended for reuse, as they may start to tear and fail but the rest of the parts can be disinfected and reused. We provide a set of guidelines to build a low-cost 3D printed solution for an effective PAPR system and describe the procedures to validate it to comply with the biosafety level 3 requirements. We have validated the prototype of PROPER unit for air flow, ISO class cleanliness level, oxygen and carbon-dioxide gas concentrations during exhalation, and present here these results for illustration. We demonstrate that the area inside the hood is more than 200 times cleaner than the external ambient without the operator and more than 175 times with the operator and in an aerosol exposed environment. We also include the procedure to clean and disinfect the equipment for reuse. PROPER may be a useful addition to provide protection to HCPs against the SARS-CoV-2 virus or other potential future viral diseases that are transmitted aerially.


Electric field mediated separation of water-ethanol mixtures in carbon-nanotubes integrated in nanoporous graphene membranes.
We investigate the influence of an applied electric field on the separation of a water-ethanol solution inside a carbon nanotube (CNT) using a series of molecular dynamics simulations. The electric field is applied at an angle θ with respect to the axis of the CNT. The study uncovers that with the application of a 'small-angle' electric field (e.g. smaller θ), the water molecules exhibit preferential occupancy inside the CNT, whereas the application of the same electric field at a 'wide-angle' mode (e.g. higher θ) fills the CNT with ethanol molecules in place of water. Remarkably, the direction of the electric field plays a pivotal role because the field exerts a contrasting influence on the behaviours of the water and ethanol molecules. The water dipoles are favourably aligned at small values of θ creating an ordered water structure inside the CNT. Increasing θ disrupts the water dipole orientation and leads to the preferential occupancy of the CNT by ethanol molecules. An in-depth analysis on the simulated systems unveil that, at lower values of θ, multiple layers of water molecules are physically adsorbed near the CNT walls, which is found to diminish as θ is increased. In comparison, at higher magnitudes of θ, the ethanol molecules are preferentially adsorbed inside the CNT. The average interaction energy per ethanol (water) molecule is found to increase (reduce) when θ is monotonically increased, which can be ascribed to the increase (decrease) in the intermolecular hydrogen bonding capacity of the ethanol (water) molecules at larger values of θ. Consequently, inside the CNT, the average occupancy of water molecules decreases and ethanol molecules increases, as θ is monotonically increased, leading to the separation of the ethanol-water mixture. The proposed methodology can convert an equimolar mixture (1 : 1) of ethanol-water into a concentrated one (14 : 1) when the electric field is applied orthogonal to the axis of the CNT. The separation efficiency is found to improve with an increase in the intensity of the externally applied electric field.


Surface plasmon resonance biosensors for simultaneous monitoring of amyloid-beta oligomers and fibrils and screening of select modulators.
Oligomeric amyloid-beta (Aβ) peptides are considered as the most toxic species in Alzheimer's disease (AD). Monitoring of the Aβ aggregation profiles is critical for elucidating the oligomer toxicity and may serve as a therapeutic target for AD. By immobilizing the capture antibodies of A11 and OC that are specific to the oligomers and fibrils, respectively, in separate fluidic channels, a novel surface plasmon resonance (SPR) biosensor was designed for monitoring the oligomeric and fibrillar species of Aβ(1-42) simultaneously. The influence of curcumin, Cu(2+) and methylene blue on the amount of toxic oligomers and fibrils was evaluated. The half maximal inhibitory concentration (IC50) of curcumin and methylene blue was determined. The formation of Aβ fibrils was also validated by the thioflavin T (ThT) fluorescence assay. The results demonstrate the utility of SPR as an analytical tool for rapid and comprehensive monitoring of Aβ aggregation and screening of Aβ modulators.


Cryopreservation of Cyrtopodium hatschbachii Pabst (Orchidaceae) immature seeds by encapsulation-dehydration.
The aim of the present study was to investigate the efficiency of the encapsulation-dehydration technique for cryopreservation of Cyrtopodium hastchbachii Pabst seeds. Immature seeds of this species were cryopreserved by an encapsulation-dehydration technique. Seeds of five immature pods, 120 days after pollination, were encapsulated in 3% calcium alginate matrix and pretreated in liquid medium supplemented with 0.08 M sucrose (24 h), 0.15 M sucrose (24 h), 0.25 M sucrose (48 h), 0.5 M sucrose (24 h) and 0.75 M sucrose (24 h) in shaker at 60 rpm. Alginate beads were dehydrated 5 h in silicagel and immersed in liquid nitrogen for 12 h. Cryopreserved beads were thawed at 30 degrees C for 1 min, rehydrated using the same liquid mediums [0.75 M sucrose (24 h), 0.5 M sucrose (24 h), 0.25 M sucrose (48 h) and 0.15 M sucrose (24 h)] and cultivated in half strength Murashige & Skoog medium (1962) with the addition of 2 g/L activated charcoal. Sixty four percent of seeds survived and developed into acclimatized plants after being cryopreserved. In this work, the encapsulation-dehydration technique was employed for first time in Cyrtopodium hatschbachii.


A comparison of MEmory Specificity Training (MEST) to education and support (ES) in the treatment of recurrent depression: study protocol for a cluster randomised controlled trial.
Depression is a debilitating mental health problem that tends to run a chronic, recurrent course. Even when effectively treated, relapse and recurrence rates remain high. Accordingly, interventions need to focus not only on symptom reduction, but also on reducing the risk of relapse by targeting depression-related disturbances that persist into remission. We are addressing this need by investigating the efficacy, acceptability and feasibility of a MEmory Specificity Training (MEST) programme, which directly targets an enduring cognitive marker of depression - reduced autobiographical memory specificity. Promising pilot data suggest that training memory specificity ameliorates this disturbance and reduces depressive symptoms. A larger, controlled trial is now needed to examine the efficacy of MEST. This trial compares MEST to an education and support (ES) group, with an embedded mechanism study. In a single blind, parallel cluster randomised controlled trial, 60 depressed individuals meeting diagnostic criteria for a current major depressive episode will be recruited from the community and clinical services. Using a block randomisation procedure, groups of 5 to 8 participants will receive five weekly sessions of MEST (n = 30) or education and support (n = 30). Participants will be assessed immediately post-treatment, and at 3- and 6-months post-treatment (MEST group only for 6-month follow-up). Depressive symptoms at 3-month follow-up will be the primary outcome. Secondary outcomes will be change in depressive status and memory specificity at post-treatment and 3-months. The 6-month follow-up of the MEST group will allow us to examine whether treatment gains are maintained. An explanatory question will examine variables mediating improvement in depression symptoms post-treatment and at 3-month follow-up. This trial will allow us to investigate the efficacy of MEST, whether treatment gains are maintained, and the mechanisms of change. Evidence will be gathered regarding whether this treatment is feasible and acceptable as a low-intensity intervention. If efficacy can be demonstrated, the results will support MEST as a treatment for depression and provide the foundation for a definitive trial. NCT01882452 (ClinicalTrials.gov), registered on 18 June 2013.


ULTRAVIOLET ABSORPTION IN EPIDERMOID CANCER CELLS.
The "excessive functional activity" of some cancer cells first found by Caspersson has been observed in fixed, stained smears of cervical epidermoid carcinomas from four patients. Preliminary results suggest that there may be a characteristic difference between the absorption profiles of some epidermoid cancer cells and other cells found in cytological smears. It is our belief that with an appropriate electronic scanning system such cells can be detected by measurements of their absorptions at two different wavelengths. However, the effect on the absorptions of cells with abnormalities other than cancer, and whether every epidermoid carcinoma will contain such cells, must yet be determined.


Characterization of bone metastases from rapid autopsies of prostate cancer patients.
Bone is the most common metastatic site for prostate cancer, and osseous metastases are the leading cause of morbidity from this disease. Recent autopsy studies prove that 100% of men who die of prostate cancer have bone involvement. Understanding the biology of prostate cancer and its evolution to an incurable androgen-independent phenotype requires an understanding of the genetic and cellular alterations that lead to the seeding and proliferation of tumor foci in bone, as well as the microenvironment in which these metastases arise. No intensive studies, however, have been conducted on osseous metastatic tissues from patients with metastatic prostate cancer due to lack of access to such tissues for profiling and other research. We show, for the first time, a reproducible methodology to obtain high quality clinical tumor tissues metastatic to the bone. This technique allowed the procurement of viable metastatic tumor tissue from involved bones in 13 recent autopsies conducted at the University of Michigan and analyzed the gene expression of these tissues using real-time PCR and microarrays. We present here the discovery of nonossified bone metastases from multiple patients with advanced prostate cancer and their subsequent characterization and comparison to nonosseous metastases from the same patients. This represents a versatile and practical approach that may be employed to characterize the steps in metastasis and the phenotypic characteristics of osseous metastasis of prostate cancer and to profile RNA, DNA, and cDNA from tumor samples metastatic to the bone.


Raman spectroscopy reveals LPS-induced changes of biomolecular composition in monocytic THP-1 cells in a label-free manner.
The human innate immune system is able to recognize pathogen-associated molecular patterns like lipopolysaccharides (LPS) leading to the activation of signal cascades and the release of different cytokines. Activation of the immune cells can be assessed in different ways which are either indirect (ELISA of cytokine release), require staining protocols (flow cytometry) or lysis of the cells (mRNA analysis). Here, Raman spectroscopy as a non-destructive spectroscopic method is presented to enable direct and label-free monitoring of changes in cellular metabolism, biomolecular composition as well as morphology. Exemplarily, the potential of Raman spectroscopy is presented for the characterization of LPS-stimulation of monocytic THP-1 cells over a time course of 16 h. The cell culture stimulation model is characterized using gene transcription and expression of the two cytokines TNFα and IL-1β. After 1 h, 3 h, 8 h and 16 h specific Raman spectroscopic fingerprints are generated which encode cell activation pattern after TLR4 stimulation. Most prevalent changes in the spectra occur after 8 h, but slight differences are already detectable after 1 h. Spatially highly resolved Raman scans are used to generate false-color Raman images which provide spatial information of the biochemical state of the cells and changes over time. One of the most significant observed differences is an increase in Raman signal from DNA/RNA content in LPS-stimulated cells when compared to unstimulated cells. The systematic assignment of Raman spectroscopic profiles of LPS-activated cells to cellular activation assessed by cytokine gene transcription and expression opens new ways for label-free and direct immunological studies of specific pathogen recognizing receptors and their downstream signaling pathways.


ABLs and TMKs are co-receptors for extracellular auxin.
Extracellular perception of auxin, an essential phytohormone in plants, has been debated for decades. Auxin-binding protein 1 (ABP1) physically interacts with quintessential transmembrane kinases (TMKs) and was proposed to act as an extracellular auxin receptor, but its role was disputed because abp1 knockout mutants lack obvious morphological phenotypes. Here, we identified two new auxin-binding proteins, ABL1 and ABL2, that are localized to the apoplast and directly interact with the extracellular domain of TMKs in an auxin-dependent manner. Furthermore, functionally redundant ABL1 and ABL2 genetically interact with TMKs and exhibit functions that overlap with those of ABP1 as well as being independent of ABP1. Importantly, the extracellular domain of TMK1 itself binds auxin and synergizes with either ABP1 or ABL1 in auxin binding. Thus, our findings discovered auxin receptors ABL1 and ABL2 having functions overlapping with but distinct from ABP1 and acting together with TMKs as co-receptors for extracellular auxin.


A c-di-GMP-Modulating Protein Regulates Swimming Motility of Burkholderia cenocepacia in Response to Arginine and Glutamate.
Burkholderia cenocepacia is an opportunistic bacterium that can thrive in different environments, including the amino acid-rich mucus of the cystic fibrosis (CF) lung. B. cenocepacia responds to the nutritional conditions that mimic the CF sputum by increasing flagellin expression and swimming motility. Individual amino acids also induce swimming but not flagellin expression. Here, we show that modulation of the second messenger cyclic dimeric guanosine monophosphate (c-di-GMP) levels by the PAS-containing c-di-GMP phosphodiesterase, BCAL1069 (CdpA), regulates the swimming motility of B. cenocepacia K56-2 in response to CF sputum nutritional conditions. Heterologous expression of WspR, a diguanylate cyclase, in B. cenocepacia K56-2 caused an increase in c-di-GMP levels and reduced swimming motility but did not affect flagellin expression or flagellar biosynthesis. After insertional mutagenesis of 12 putative genes encoding c-di-GMP metabolizing enzymes, one mutant of the locus BCAL1069 (cdpA), exhibited decreased swimming motility independent of flagellin expression in CF sputum nutritional conditions and an increase in intracellular c-di-GMP levels. The reduced swimming motility phenotype of the BCAL1069 mutant was observed in the presence of arginine and glutamate, but not of histidine, phenylalanine, or proline. The B. cenocepacia CdpA was also found to be involved in regulation of protease activity but not in biofilm formation. Altogether, these results highlight a role of B. cenocepacia BCAL1069 (CdpA) in sensing the nutritional conditions of the CF sputum and eliciting a pathogenic response that includes swimming motility toward amino acids and an increase in protease activity.


Epigenetics: relevance and implications for public health.
Improved understanding of the multilayer regulation of the human genome has led to a greater appreciation of environmental, nutritional, and epigenetic risk factors for human disease. Chromatin remodeling, histone tail modifications, and DNA methylation are dynamic epigenetic changes responsive to external stimuli. Careful interpretation can provide insights for actionable public health through collaboration between population and basic scientists and through integration of multiple data sources. We review key findings in environmental epigenetics both in human population studies and in animal models, and discuss the implications of these results for risk assessment and public health protection. To ultimately succeed in identifying epigenetic mechanisms leading to complex phenotypes and disease, researchers must integrate the various animal models, human clinical approaches, and human population approaches while paying attention to life-stage sensitivity, to generate effective prescriptions for human health evaluation and disease prevention.


Multiscale penalized weighted least-squares sinogram restoration for low-dose x-ray computed tomography.
In this paper, we propose a novel multiscale penalized weighted least-squares (PWLS) method for restoration of low-dose computed tomography (CT) sinogram. The method utilizes wavelet transform for the multiscale or multiresolution analysis on the sinogram. Specifically, the Mallat-Zhong's wavelet transform is applied to decompose the sinogram to different resolution levels. At each decomposed resolution level, a PWLS criterion is applied to restore the noise-contaminated wavelet coefficients, where the penalty is adaptive to each resolution scale and the weight is updated by an exponential relationship between the data variance and mean at each scale and location. The proposed PWLS method is based on the observations that 1) noise in the CT sinogram after logarithm transform and calibration can be modeled as signal-dependent variables and the sample variance depends on the sample mean by an exponential relationship; and 2) noise reduction can be more effective when it is adaptive to different resolution levels. The effectiveness of the proposed multiscale PWLS method is validated by both computer simulations and experimental studies. The gain by multiscale approach over single scale means is quantified by noise-resolution tradeoff measures.


Unraveling the mystery of the hygiene hypothesis through Helicobacter pylori infection.
Epidemiological studies have revealed an inverse association between Helicobacter pylori infection and the incidence of allergic asthma. This association is consistent with the hygiene hypothesis, which posits that exposure to microbes early in life prevents the later development of allergic diseases, and has been reproduced in mouse models of asthma. In this issue of the JCI, Oertli and colleagues report that H. pylori infection in neonates elicits tolerogenic DCs that produce IL-18, which drive the generation of Tregs that subsequently protect the mice from allergic asthma. This finding strengthens the intriguing link between pathogen exposure and allergic disease.


Control of ethylene synthesis by expression of a bacterial enzyme in transgenic tomato plants.
Synthesis of the phytohormone ethylene is believed to be essential for many plant developmental processes. The control of ripening in climacteric fruits and vegetables is among the best characterized of these processes. One approach to reduce ethylene synthesis in plants is metabolism of its immediate precursor, 1-aminocyclopropane-1-carboxylic acid (ACC). Soil bacteria containing an enzyme, ACC deaminase, were identified by their ability to grow on ACC as a sole nitrogen source. The gene encoding ACC deaminase was cloned and introduced into tomato plants. Reduction in ethylene synthesis in transgenic plants did not cause any apparent vegetative phenotypic abnormalities. However, fruits from these plants exhibited significant delays in ripening, and the mature fruits remained firm for at least 6 weeks longer than the nontransgenic control fruit. These results indicated that ACC deaminase is useful for examining the role of ethylene in many developmental and stress-related processes in plants as well as for extending the shelf life of fruits and vegetables whose ripening is mediated by ethylene.


Determination of the normative values of the masseter muscle by single-fiber electromyography in myasthenia gravis patients.
The purpose of this study is to obtain normative values of the masseter muscle of myasthenia gravis (MG) patients and healthy volunteers by single-fiber electromyography (SFEMG). Stimulation of SFEMG in the masseter muscle was studied in 15 healthy volunteers (men 8, women 7; mean age 40.2, range 21-77) and 30 patients affected by MG (men 16, women 14; mean age 42.8, range 12-75). The mean consecutive difference (MCD) of the individual fiber and the mean MCD per study were determined in the normal group. We recommend the upper normal limit for the individual fibers of jitter and the mean MCD per study in the healthy Chinese adults of 33 µs and 22 µs respectively. Furthermore, in the MG group, the percentage of jitter > upper normal limit jitter and the impulse blocking percentage were detected, which were all significantly different compared to the normal control group (P < 0.01). The overall sensitivity was 90%, with the abnormality in 6 of the 9 ocular MG patients and 100% abnormality in the generalized MG patients. The masseter muscle SFEMG has a high degree of sensitivity. The masseter should be considered for SFEMG in the diagnosis of MG, and added routinely to the tested muscles.


Differentiation of the maize subgenomes by genome dominance and both ancient and ongoing gene loss.
Ancient tetraploidies are found throughout the eukaryotes. After duplication, one copy of each duplicate gene pair tends to be lost (fractionate). For all studied tetraploidies, the loss of duplicated genes, known as homeologs, homoeologs, ohnologs, or syntenic paralogs, is uneven between duplicate regions. In maize, a species that experienced a tetraploidy 5-12 million years ago, we show that in addition to uneven ancient gene loss, the two complete genomes contained within maize are differentiated by ongoing fractionation among diverse inbreds as well as by a pattern of overexpression of genes from the genome that has experienced less gene loss. These expression differences are consistent over a range of experiments quantifying RNA abundance in different tissues. We propose that the universal bias in gene loss between the genomes of this ancient tetraploid, and perhaps all tetraploids, is the result of selection against loss of the gene responsible for the majority of total expression for a duplicate gene pair. Although the tetraploidy of maize is ancient, biased gene loss and expression continue today and explain, at least in part, the remarkable genetic diversity found among modern maize cultivars.


Effects of tryptophan depletion on brain potential correlates of episodic memory retrieval.
Neuropsychological impairments in depressive illness may be secondary to proposed serotonergic abnormalities. Acute tryptophan depletion (ATD) in healthy subjects impairs episodic memory, but the mechanism of this is unclear. To examine the effects of ATD on the neural correlates of episodic memory retrieval in healthy subjects. Fourteen healthy men were given an amino acid cocktail drink with or without tryptophan, in a double blind, crossover design. Event related potentials (ERPs) were recorded during a well-validated episodic memory task performed 5 h after drink ingestion. Subjects listened to words spoken in a male or female voice. At test, old and new words were presented visually; subjects judged whether words were old or new, and if old, the gender of the voice at study. ATD led to an 84+/-5% reduction in plasma free tryptophan concentrations, and significantly impaired episodic memory recall. ERP recordings demonstrated previously reported left parietal and right frontal "old/new" differences for ERPs to items associated with accurate episodic memory retrieval versus correctly rejected new items. ATD increased ERP voltage between 500 and 1400 ms post-stimulus particularly over posterior regions of the scalp, but there was no interaction with item type. Topographical analysis of the old/new difference revealed no significant treatment by site interaction. ATD impairs episodic memory recall with no effect on the magnitude or topography of the neural correlates of retrieval in healthy subjects. This suggests that the effects of ATD on recall may reflect an impairment of memory encoding and/or consolidation.


Implications of the plastid genome sequence of typha (typhaceae, poales) for understanding genome evolution in poaceae.
Plastid genomes of the grasses (Poaceae) are unusual in their organization and rates of sequence evolution. There has been a recent surge in the availability of grass plastid genome sequences, but a comprehensive comparative analysis of genome evolution has not been performed that includes any related families in the Poales. We report on the plastid genome of Typha latifolia, the first non-grass Poales sequenced to date, and we present comparisons of genome organization and sequence evolution within Poales. Our results confirm that grass plastid genomes exhibit acceleration in both genomic rearrangements and nucleotide substitutions. Poaceae have multiple structural rearrangements, including three inversions, three genes losses (accD, ycf1, ycf2), intron losses in two genes (clpP, rpoC1), and expansion of the inverted repeat (IR) into both large and small single-copy regions. These rearrangements are restricted to the Poaceae, and IR expansion into the small single-copy region correlates with the phylogeny of the family. Comparisons of 73 protein-coding genes for 47 angiosperms including nine Poaceae genera confirm that the branch leading to Poaceae has significantly accelerated rates of change relative to other monocots and angiosperms. Furthermore, rates of sequence evolution within grasses are lower, indicating a deceleration during diversification of the family. Overall there is a strong correlation between accelerated rates of genomic rearrangements and nucleotide substitutions in Poaceae, a phenomenon that has been noted recently throughout angiosperms. The cause of the correlation is unknown, but faulty DNA repair has been suggested in other systems including bacterial and animal mitochondrial genomes.


Learning to understand others' actions.
Despite nearly two decades of research on mirror neurons, there is still much debate about what they do. The most enduring hypothesis is that they enable 'action understanding'. However, recent critical reviews have failed to find compelling evidence in favour of this view. Instead, these authors argue that mirror neurons are produced by associative learning and therefore that they cannot contribute to action understanding. The present opinion piece suggests that this argument is flawed. We argue that mirror neurons may both develop through associative learning and contribute to inferences about the actions of others.


Non-Nucleoside Reverse Transcriptase Inhibitor-Based Antiretroviral Therapy in Perinatally HIV-Infected, Treatment-Naïve Adolescents in Asia.
About a third of untreated, perinatally HIV-infected children reach adolescence. We evaluated the durability and effectiveness of non-nucleoside reverse-transcriptase inhibitor (NNRTI)-based antiretroviral therapy (ART) in this population. Data from perinatally HIV-infected, antiretroviral-naïve patients initiated on NNRTI-based ART aged 10-19 years who had ≥6 months of follow-up were analyzed. Competing risk regression was used to assess predictors of NNRTI substitution and clinical failure (World Health Organization Stage 3/4 event or death). Viral suppression was defined as a viral load <400 copies/mL. Data from 534 adolescents met our inclusion criteria (56.2% female; median age at treatment initiation 11.8 years). After 5 years of treatment, median height-for-age z score increased from -2.3 to -1.6, and median CD4+ cell count increased from 131 to 580 cells/mm(3). The proportion of patients with viral suppression after 6 months was 87.6% and remained >80% up to 5 years of follow-up. NNRTI substitution and clinical failure occurred at rates of 4.9 and 1.4 events per 100 patient-years, respectively. Not using cotrimoxazole prophylaxis at ART initiation was associated with NNRTI substitution (hazard ratio [HR], 1.5 vs. using; 95% confidence interval [CI] = 1.0-2.2; p = .05). Baseline CD4+ count ≤200 cells/mm(3) (HR, 3.3 vs. >200; 95% CI = 1.2-8.9; p = .02) and not using cotrimoxazole prophylaxis at ART initiation (HR, 2.1 vs. using; 95% CI = 1.0-4.6; p = .05) were both associated with clinical failure. Despite late ART initiation, adolescents achieved good rates of catch-up growth, CD4+ count recovery, and virological suppression. Earlier ART initiation and routine cotrimoxazole prophylaxis in this population may help to reduce current rates of NNRTI substitution and clinical failure.


Mechanism of aminoglycoside antibiotic kinase APH(3')-IIIa: role of the nucleotide positioning loop.
The aminoglycoside antibiotic resistance kinases (APHs) and the Ser/Thr/Tyr protein kinases share structural and functional homology but very little primary sequence conservation (<5%). A region of structural, but not amino acid sequence, homology is the nucleotide positioning loop (NPL) that closes down on the enzyme active site upon binding of ATP. This loop region has been implicated in facilitating phosphoryl transfer in protein kinases; however, there is no primary sequence conservation between APHs and protein kinases in the NPL. There is an invariant Ser residue in all APH NPL regions, however. This residue in APH(3')-IIIa (Ser27), an enzyme widespread in aminoglycoside-resistant Enterococci, Streptococci, and Staphylococci, directly interacts with the beta-phosphate of ATP through the Ser hydroxymethyl group and the amide hydrogen in the 3D structure of the enzyme. Mutagenesis of this residue to Ala and Pro supported a role for the Ser amide hydrogen in nucleotide capture and phosphoryl transfer. A molecular model of the proposed dissociative transition state, which is consistent with all of the available mechanistic data, suggested a role for the amide of the adjacent Met26 in phosphoryl transfer. Mutagenesis studies confirmed the importance of the amide hydrogen and suggest a mechanism where Ser27 anchors the ATP beta-phosphate facilitating bond breakage with the gamma-phosphate during formation of the metaphosphate-like transition, which is stabilized by interaction with the amide hydrogen of Met26. The APH NPL therefore acts as a lever, promoting phosphoryl transfer to the aminoglycoside substrate, with the biological outcome of clinically relevant antibiotic resistance.


Patient-physician discordance in fibromyalgia.
Discordance between patients' and physicians' health perceptions and satisfaction with the office visit in fibromyalgia (FM) has not been examined. We investigated this phenomenon to identify demographic, clinical, and psychosocial factors associated with patient-physician discordance on physical functioning, well being, and satisfaction with the office visit. A sample of 182 women were examined by a rheumatologist to confirm the FM diagnosis. Patients and physicians independently completed the Patient-Physician Discordance Scale to assess perceptions of health and satisfaction with the office visit. Patients also completed questionnaires pertaining to sociodemographics, social support, disability, perceived stress, and psychological distress following the office visit. Separate generalized estimating equations with forward selection, controlling for the possible dependence of outcomes among patients of same physician, were modeled for each measure of discordance. The highest discordance score was on satisfaction with the office visit; physicians systematically underestimated patients' level of satisfaction. Higher levels of satisfaction with social support (p < 0.02) and more psychological distress (p < 0.03) were marginally associated with greater discordance on physical functioning. Higher levels of satisfaction with social support (p < 0.003), younger age (p < 0.02), and lower disability (p < 0.03) were associated with greater discordance on well being. More sexual abuse (p < 0.01) was significantly associated with more discordance on satisfaction with the office visit. There is a gap between what patients with FM and rheumatologists examining them experience during the office visit. Psychosocial factors contribute to our understanding of discordance on physical functioning, well being, and satisfaction.


SMAD4 Is Essential for Human Cardiac Mesodermal Precursor Cell Formation.
Understanding stage-specific molecular mechanisms of human cardiomyocyte (CM) progenitor formation and subsequent differentiation are critical to identify pathways that might lead to congenital cardiovascular defects and malformations. In particular, gene mutations in the transforming growth factor (TGF)β superfamily signaling pathways can cause human congenital heart defects, and murine loss of function studies of a central component in this pathway, Smad4, leads to early embryonic lethality. To define the role of SMAD4 at the earliest stages of human cardiogenesis, we generated SMAD4 mutant human embryonic stem cells (hESCs). Herein, we show that the loss of SMAD4 has no effect on hESC self-renewal, or neuroectoderm formation, but is essential for the formation of cardiac mesoderm, with a subsequent complete loss of CM formation during human ES cell cardiogenesis. Via transcriptional profiling, we show that SMAD4 mutant cell lines fail to generate cardiac mesodermal precursors, clarifying a role of NODAL/SMAD4 signaling in cardiac mesodermal precursor formation via enhancing the expression of primitive streak genes. Since SMAD4 relative pathways have been linked to congenital malformations, it will become of interest to determine whether these may due, in part, to defective cell fate decision during cardiac mesodermal precursor formation. Stem Cells 2018 Stem Cells 2019;37:216-225.


No association between dietary vitamin K intake and fracture risk in chinese community-dwelling older men and women: a prospective study.
Data on the association between dietary vitamin K intake and fracture risk are limited among Chinese. This study examined such an association in community-dwelling elderly in Hong Kong. We present data from 2,944 subjects (1,605 men, 1,339 women) who participated in a prospective cohort study. Baseline dietary intakes of energy, protein, calcium, vitamin D, and vitamin K were assessed using a food-frequency questionnaire. Data on incident hip fracture and nonvertebral fracture during a median of 6.9 follow-up years were collected from a hospital database. Cox regression analyses were performed with adjustments for age, education attainment, smoking status, alcohol use, body mass index, hip bone mineral density, physical activity, use of calcium supplement, and energy-adjusted nutrient intakes. There were 29 (1.8 %) men and 19 (1.4 %) women with incident hip fractures and 97 (6.0 %) men and 88 (6.6 %) women with nonvertebral fractures. The median (interquartile range) of dietary vitamin K intake was 241.8 (157.5-360.8) and 238.9 (162.4-343.6) μg/day in men and women, respectively. Similar dietary vitamin K intakes were observed between subjects with hip or nonvertebral fractures and subjects without hip or nonvertebral fractures. In both men and women, dietary vitamin K intake was not associated with fracture risks at all measured sites in either crude or adjusted models. In Chinese community-dwelling elderly, hip or nonvertebral fracture risk was not associated with dietary vitamin K intake. The high dietary vitamin K intake of the studied group may have limited the ability to detect the association between vitamin K intake and fracture risk.


Contribution of adipose triglyceride lipase and hormone-sensitive lipase to lipolysis in hMADS adipocytes.
Lipolysis is the catabolic pathway by which triglycerides are hydrolyzed into fatty acids. Adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) have the capacity to hydrolyze in vitro the first ester bond of triglycerides, but their respective contributions to whole cell lipolysis in human adipocytes is unclear. Here, we have investigated the roles of HSL, ATGL, and its coactivator CGI-58 in basal and forskolin-stimulated lipolysis in a human white adipocyte model, the hMADS cells. The hMADS adipocytes express the various components of fatty acid metabolism and show lipolytic capacity similar to primary cultured adipocytes. We show that lipolysis and fatty acid esterification are tightly coupled except in conditions of stimulated lipolysis. Immunocytochemistry experiments revealed that acute forskolin treatment promotes HSL translocation from the cytosol to small lipid droplets and redistribution of ATGL from the cytosol and large lipid droplets to small lipid droplets, resulting in enriched colocalization of the two lipases. HSL or ATGL overexpression resulted in increased triglyceride-specific hydrolase capacity, but only ATGL overexpression increased whole cell lipolysis. HSL silencing had no effect on basal lipolysis and only partially reduced forskolin-stimulated lipolysis. Conversely, silencing of ATGL or CGI-58 significantly reduced basal lipolysis and essentially abolished forskolin-stimulated lipolysis. Altogether, these results suggest that ATGL/CGI-58 acts independently of HSL and precedes its action in the sequential hydrolysis of triglycerides in human hMADS adipocytes.


Variation and transgression of aggressiveness among two Gibberella zeae crosses developed from highly aggressive parental isolates.
Gibberella zeae (anamorph: Fusarium graminearum) is the most common cause of Fusarium head blight (FHB) of wheat (Triticum aestivum) worldwide. Aggressiveness is the most important fungal trait affecting disease severity and stability of host resistance. Objectives were to analyze in two field experiments (i) segregation for aggressiveness among 120 progenies from each of two crosses of highly aggressive parents and (ii) stability of FHB resistance of seven moderately to highly resistant winter wheat cultivars against isolates varying for aggressiveness. Aggressiveness was measured as FHB severity per plot, Fusarium exoantigen absorbance, and deoxynivalenol content. In the first experiment, mean FHB ratings were 20 to 49% across environments and progeny. Significant genotypic variation was detected in both crosses (P < 0.01). Isolate-environment interaction explained approximately half of the total variance. Two transgressive segregants were found in cross B across environments. Traits were significantly (P < 0.05) intercorrelated. In the second experiment, despite significant (P < 0.05) genotypic variance for cultivar and isolate, no significant (P > 0.05) interaction was observed for any trait. In conclusion, progeny of highly aggressive parents might exhibit increased aggressiveness due to recombination and may, therefore, adapt nonspecifically to increased quantitative host resistance.


Whole-body vibration effects on bone mineral density in women with or without resistance training.
Whole-body vibration exposure may translate into improved bone mass in young adult women. The primary focus of this study was to examine the effects of graded whole-body vibration or vibration exposure plus resistance training on bone mineral density (BMD), hematological measures for bone remodeling, and exercise metabolism in young women. There were 51 healthy active women [mean (SD) age, 21.02 (3.39) yr; height, 165.66 (6.73) cm; body mass 66.54 (13.39) kg] who participated in the intervention. Subjects were randomly assigned to whole-body vibration (WBV), whole-body vibration plus resistance training (WBV+RT), or control (CONT) groups for 16 wk. A repeated-measure ANOVA found no significant (P < 0.05) group differences in BMD at the completion of 16 wk. A significant within group change was apparent for the WBV (2.7% femoral neck) and WBV+RT (femoral neck 1.9%; vertebra 0.98%). WBV and WBV+RT experienced a significant (P < 0.05) 60% and 58% increase in adiponectin, 48% and 30% in transforming growth factor-beta1, and 17% and 34% in nitric oxide with an accompanying 50% and 36% decrease in osteopontin, 19% and 34% in interleukin-1beta, and 38% and 39% in tumor necrosis factor-alpha. The results indicate graded whole-body vibration exposure may be effective in improving BMD by increasing bone deposition while also decreasing bone resorption. Whole-body vibration may also provide an efficient stratagem for young women to achieve peak bone mass and help stave off osteoporosis later in life and provide a novel form of physical training.


Boundary cap cells constrain spinal motor neuron somal migration at motor exit points by a semaphorin-plexin mechanism.
In developing neurons, somal migration and initiation of axon outgrowth often occur simultaneously and are regulated in part by similar classes of molecules. When neurons reach their final destinations, however, somal translocation and axon extension are uncoupled. Insights into the mechanisms underlying this process of disengagement came from our study of the behaviour of embryonic spinal motor neurons following ablation of boundary cap cells. These are neural crest derivatives that transiently reside at motor exit points, central nervous system (CNS):peripheral nervous system (PNS) interfaces where motor axons leave the CNS. In the absence of boundary cap cells, motor neuron cell bodies migrate along their axons into the periphery, suggesting that repellent signals from boundary cap cells regulate the selective gating of somal migration and axon outgrowth at the motor exit point. Here we used RNA interference in the chick embryo together with analysis of null mutant mice to identify possible boundary cap cell ligands, their receptors on motor neurons and cytoplasmic signalling molecules that control this process. We demonstrate that targeted knock down in motor neurons of Neuropilin-2 (Npn-2), a high affinity receptor for class 3 semaphorins, causes their somata to migrate to ectopic positions in ventral nerve roots. This finding was corroborated in Npn-2 null mice, in which we identified motor neuron cell bodies in ectopic positions in the PNS. Our RNA interference studies further revealed a role for Plexin-A2, but not Plexin-A1 or Plexin-A4. We show that chick and mouse boundary cap cells express Sema3B and 3G, secreted semaphorins, and Sema6A, a transmembrane semaphorin. However, no increased numbers of ectopic motor neurons are found in Sema3B null mouse embryos. In contrast, Sema6A null mice display an ectopic motor neuron phenotype. Finally, knockdown of MICAL3, a downstream semaphorin/Plexin-A signalling molecule, in chick motor neurons led to their ectopic positioning in the PNS. We conclude that semaphorin-mediated repellent interactions between boundary cap cells and immature spinal motor neurons regulates somal positioning by countering the drag exerted on motor neuron cell bodies by their axons as they emerge from the CNS at motor exit points. Our data support a model in which BC cell semaphorins signal through Npn-2 and/or Plexin-A2 receptors on motor neurons via a cytoplasmic effector, MICAL3, to trigger cytoskeletal reorganisation. This leads to the disengagement of somal migration from axon extension and the confinement of motor neuron cell bodies to the spinal cord.


An alternative treatment regimen of advanced seminoma with carboplatin, etoposide, and bleomycin instead of cisplatin-based therapy.
Cisplatin-based therapy is associated with acute and late toxicities. Therefore, a potentially less toxic carboplatin-based regimen was evaluated in patients with advanced seminoma. Eighteen patients with advanced seminoma were treated on outpatient basis with carboplatin (AUC5) at day 1, etoposide (100 mgm(-2)) at days 1-5, and bleomycin (30 IU) at day 2 (CEB). Treatment was 3-weekly for a total of 4 cycles. Outcome and toxicities were analyzed. Median follow-up was 4 years and 7.5 months. Five-year progression-free survival was 86.6% (95% confidence interval (CI), 70.6%-100%), 5-year overall survival 100%, and 10-year overall survival 85% (95% CI, 63.3%-100%); 39% of all patients reached complete remission. Two patients underwent adjuvant treatment. Two patients relapsed; 1 is in ongoing remission 4 years after salvage therapy, the other died almost 6 years after CEB-therapy, despite multiple lines of salvage therapy. The main acute toxicity observed was hematologic. No late cardiovascular events or secondary malignancies were noted. CEB treatment is effective in advanced seminoma, showing minor toxicity. Progression-free and overall survival rates at 5 and 10 years are comparable to those achieved with cisplatin-based therapy. This indicates that carboplatin-combination therapy might be a good alternative to cisplatin-based therapy in the treatment of advanced seminomas.