Fluorescent BODIPY-based Zn(II) complex as a molecular probe for selective detection of neurofibrillary tangles in the brains of Alzheimer's disease patients.
We have developed a new fluorescent binuclear Zn(II) complex for the detection of neurofibrillary tangles (NFTs) of hyperphosphorylated tau proteins, a representative hallmark of Alzheimer's disease (AD). The probe 1 incorporates a fluorescent BODIPY unit and two Zn(II)-2,2'-dipicolylamine (Dpa) complexes as a binding site for phosphorylated amino acid residues. Using fluorescence titration to evaluate the binding and sensing properties of 1 toward several phosphorylated peptide segments derived from hyperphosphorylated tau protein, we found that 1 binds preferentially to peptides presenting phosphorylated groups at the i and i+4 positions with dissociation constants (K(d)) in the micromolar range. Fluorescence titration with an artificially prepared aggregate of the phosphorylated tau protein (p-Tau) revealed that 1 binds strongly to p-Tau (EC(50) = 9 nM). In contrast, the interactions of 1 were weaker toward artificially prepared aggregates of the nonphosphorylated tau protein (n-Tau; EC(50) = 80 nM) and Abeta(1-42) fibrils (EC(50) = 650 nM). Histological imaging of a hippocampus tissue section obtained from an AD patient revealed that 1 fluorescently visualizes deposits of NFTs and clearly discriminates between NFTs and the amyloid plaques assembled from amyloid-beta peptides, confirming our strategy toward the rational design of a molecular probe for the selective fluorescence detection of NFTs in brain tissue sections.


A pathologic study of degeneration of the rod and cone populations of the rhodopsin Pro347Leu transgenic pigs.
Transgenic pigs with rhodopsin (Pro347Leu) mutation exhibited rod-cone degeneration. We compared the pathologic characteristics of the rod degeneration versus those of the cone cells. The posterior and peripheral retinas of these transgenic pigs of age 4, 6, 8, 12, 24 and 33 weeks and normal pigs of age 4 and 8 weeks were studied by light and EM and morphometry. The pathologic changes observed in the posterior and peripheral retinas of the transgenic pigs could be conveniently described in 3 phases: I) an initial phase of rapid and extensive degeneration of the rod cells in the first 6 weeks of age; II) an acute phase of cone cell degeneration involving approximately half of the population and lingering rod degeneration in the 6 to 12 weeks of age; and III) a partial cone recovery to be followed by a chronic degenerative phase of the remaining cones cells from 12 to 33 weeks of age. Our study showed that the degenerative changes of rod cells could be differentiated from those of the cone cells. Cone and rod populations degenerated along different time schedule with different pathologic features. Hence, treatment for retinitis pigmentosa might vary with the different stages of the disease.


New Investigator and Trainee Task Force Survey on the Recruitment and Retention of Headache Specialists.
We sought to survey the New Investigators and Trainees Section (NITS) members of the American Headache Society (AHS) to better understand their exposure to headache medicine during training and to determine their perceptions and attitudes about the field and the future of headache medicine. Despite the high prevalence of headache disorders in the general population, only about 2% of neurology residents pursue headache medicine fellowships. Furthermore, there is a paucity of United Council of Neurologic Subspecialties headache specialists in the country to meet the population demands. Thus, there needs to be a focus on how to recruit and retain more headache specialists. A survey was distributed via SurveyMonkey to the NITS listserv. It remained online for 60 days, during which reminder emails were sent to members of the listserv. In addition, the survey was available on laptops at NITS-related events at an annual AHS meeting. Descriptive analyses were then conducted using SurveyMonkey and Excel. Of the 93 members of NITS, 64 of the 96 (68.8%) clicked to initiate the survey and 52.7% successfully completed it. Attendings made up the majority of respondents (62.5%), followed by fellows (10.9%), and residents (7.8%). Key highlights of the survey included the following: just under 10% reported no exposure to a headache center during any time in their training (medical school, residency, or fellowship); less than 2% had exposure to a headache center during medical school; less than half of participants reported exposure to a headache center in residency (45.3%) and during fellowship (43.4%). Having a mentor in the field, liking the patient population, and working in a headache center, 64.7%, 52.9%, and 41.2%, respectively, were the top ways in which participants became interested in headache. The journal Headache (56.9%), attendings (56.3%), and the AHS/American Academy of Neurology guidelines for migraine management (52.0%) are the resources cited as being used all/most of the time. About 82.4% strongly agree that there needs to be improved headache education for physicians of all specialties (primary care, emergency department, psychiatry); 84.4% feel that they are appreciated by their patients; 68.6% feel that there is strong support in their departments for headache; 56.9% believe that their work schedule leaves enough time for personal and family life; and 60.8% agreed that their professional life will improve in years to come. Participants agreed/strongly agreed that they like to treat the following diseases/symptoms: migraine headache (98.0%), cluster headache (92%), chronic daily headache (84%), and post-concussive syndrome (71.4%). Participants disagreed/strongly disagreed that they like to treat the following comorbid conditions/symptoms: low back pain (66.6%), dizziness (42.9%), sleep apnea (36.7%), depression (32.0%), and anxiety (32.0%). In this detailed survey on the recruitment and retention of headache specialists, the following themes emerged: mentorship and exposure to a headache center are key foundations in the young investigator/trainee experience. Young headache specialists appear positive about their field of medicine. These specialists like to treat various headache types but not necessarily some of the related comorbidities (sleep disorders, depression, anxiety, back pain, and dizziness). Finally, there was strong agreement that there needs to be improved headache education for physicians of other medical specialties.


DQB1 locus alone explains most of the risk and protection in narcolepsy with cataplexy in Europe.
Prior research has identified five common genetic variants associated with narcolepsy with cataplexy in Caucasian patients. To replicate and/or extend these findings, we have tested HLA-DQB1, the previously identified 5 variants, and 10 other potential variants in a large European sample of narcolepsy with cataplexy subjects. Retrospective case-control study. A recent study showed that over 76% of significant genome-wide association variants lie within DNase I hypersensitive sites (DHSs). From our previous GWAS, we identified 30 single nucleotide polymorphisms (SNPs) with P < 10(-4) mapping to DHSs. Ten SNPs tagging these sites, HLADQB1, and all previously reported SNPs significantly associated with narcolepsy were tested for replication. For GWAS, 1,261 narcolepsy patients and 1,422 HLA-DQB1*06:02-matched controls were included. For HLA study, 1,218 patients and 3,541 controls were included. None of the top variants within DHSs were replicated. Out of the five previously reported SNPs, only rs2858884 within the HLA region (P < 2x10(-9)) and rs1154155 within the TRA locus (P < 2x10(-8)) replicated. DQB1 typing confirmed that DQB1*06:02 confers an extraordinary risk (odds ratio 251). Four protective alleles (DQB1*06:03, odds ratio 0.17, DQB1*05:01, odds ratio 0.56, DQB1*06:09 odds ratio 0.21, DQB1*02 odds ratio 0.76) were also identified. An overwhelming portion of genetic risk for narcolepsy with cataplexy is found at DQB1 locus. Since DQB1*06:02 positive subjects are at 251-fold increase in risk for narcolepsy, and all recent cases of narcolepsy after H1N1 vaccination are positive for this allele, DQB1 genotyping may be relevant to public health policy.


A novel extract from bovine colostrum whey supports innate immune functions. II. Rapid changes in cellular immune function in humans.
To evaluate acute effects of bovine colostrum low-molecular weight fraction (CLMWF) on selected aspects of innate immune function in healthy human subjects. A placebo-controlled, double-blinded, randomized cross-over trial involving 12 healthy subjects, age 22-72, was conducted at NIS Labs during the year 2010. Placebo or 150 mg CLMWF was given orally. Blood was drawn immediately before and at 1 and 2h after consumption. A single dose of CLMWF, when compared to placebo, resulted in rapid increase in phagocytic activity of monocytes at 1h (P<0.12) and polymorphonuclear cells at 1h (P<0.08) and 2h (P<0.03) after consumption. Observations included increased numbers of CD3(+) T cells (P<0.05), and a transient reduction in circulating CD3(-)CD56(+) natural killer (NK) cells at 1h (P<0.04), returning to normal levels at 2h after consumption (P<0.96). The relative increase of NK cells from 1 to 2h after consumption was not associated with an increase in CD69 or CD25 activation markers, suggesting that new NK cells were mobilized into circulation. The increased phagocytic activity and rapid transient changes in NK cell numbers suggest that upon consumption, interaction of CLMWF with immune cells in the gut mucosa triggers immediate events with systemic consequences.


Total fluoride intake and urinary excretion in 4-year-old Iranian children residing in low-fluoride areas.
Knowledge of levels of fluoride ingestion and excretion is important in planning optimum fluoride therapy for young children. In previous literature, it has been assumed that only about one-third of ingested fluoride is excreted in young children. The aims of the present study were (a) to measure total fluoride intake, urinary fluoride excretion and fluoride balance, and (b) to investigate the effect of air temperature on fluoride intake and urinary fluoride excretion, in young children. Children (4 years old) living in a city, a small town and rural areas of Fars province, Iran, where drinking water contained 0.30-0.39 mg F/l, were invited to participate. Selection of subjects was by random sampling of kindergartens or health centres. The children were surveyed twice, once in summer and once in winter. Diet was obtained by 3 d diaries with interview. Samples of most foods and drinks were analysed for fluoride content. Ingestion of fluoride from toothpaste was estimated for each child. Each child's urine was collected over 24 h and analysed for fluoride content. Seventy-eight of the 116 volunteers completed all aspects of the study, which was conducted in 1995-6. For all children, the mean fluoride ingestion from diet was 0.390 (SD 0.122) mg/d or 0.028 (SD 0.008) mg/kg body weight per d. Fluoride ingestion from diet was higher in summer and higher in rural areas. The mean ingestion of fluoride from all sources was 0.426 (SD 0.126) mg/d and the mean fluoride urinary excretion was 0.339 (SD 0.100) mg/d. The difference between ingestion and urinary excretion was +0.087 (SD 0.143) mg, equivalent to 80% excretion. Faecal excretion was not estimated. The results indicate fluoride retention at 4 years to be much lower than previously assumed.


Community-level impacts of sanitation coverage on maternal and neonatal health: a retrospective cohort of survey data.
Access to sanitation facilities (toilets or latrines) greatly improves human health. Low community sanitation coverage may lead to increased exposure to pathogens for households both with and without a sanitation facility. We created a retrospective cohort using Demographic and Health Surveys from 1990 through 2018. Using regression with matched women as a random intercept, we assessed the association between community-level sanitation coverage and neonatal mortality (Poisson model, n=1 254 862 live births, 187 datasets), small birth size (logit model, n=1 058 843 live births, 187 datasets) and anaemia (logit model, n=1 304 626 women, 75 datasets). Among women with household sanitation, the incidence of neonatal death (incidence rate ratio: 0.85, 95% CI 0.77 to 0.93), the odds of small birth size (OR: 0.81, 95% CI 0.76 to 0.87) and anaemia (OR: 0.82, 95% CI 0.79 to 0.85) were lower for women in communities with 100% sanitation coverage compared with 1%-30% (p≤0.001 for all). There was no difference in neonatal deaths between women in communities with 31%-99% sanitation coverage compared with 1%-30% (p≥0.05). Among women without household sanitation, there were no differences in neonatal mortality by community sanitation (p≥0.05). The odds of small birth size were decreased (OR: 0.91, 95% CI 0.87 to 0.97, p=0.003) for women in communities with 61%-99% sanitation coverage compared with 1%-30%; there was no association with the other community sanitation categories (p≥0.05). The odds of anaemia were increased (OR: 1.08, 95% CI 1.06 to 1.11, p<0.001) for women living in communities with 0% sanitation coverage compared with 1%-30%, but no association with the other community sanitation categories (p≥0.05). Community sanitation coverage is associated with improved maternal and neonatal outcomes, particularly among women with household sanitation. This suggests that the impact of sanitation coverage on maternal and neonatal health is underestimated unless the community-level effects are considered.


pp32/PHAPI determines the apoptosis response of non-small-cell lung cancer.
During malignant transformation, cancer cells have to evade cell-intrinsic tumor suppressor mechanisms including apoptosis, thus acquiring a phenotype that is relatively resistant to clinically applied anticancer therapies. Molecular characterization of apoptotic signal transduction defects may help to identify prognostic markers and to develop novel therapeutic strategies. To this end we have undertaken functional analyses of drug-induced apoptosis in human non-small cell-lung cancer (NSCLC) cells. We found that primary drug resistance correlated with defects in apoptosome-dependent caspase activation in vitro. While cytochrome c-induced apoptosome formation was maintained, the subsequent activation of caspase-9 and -3 was abolished in resistant NSCLC. The addition of recombinant pp32/putative human HLA class II-associated protein (pp32/PHAPI), described as a putative tumor suppressor in prostate cancer, successfully restored defective cytochrome c-induced caspase activation in vitro. Conditional expression of pp32/PHAPI sensitized NSCLC cells to apoptosis in vitro and in a murine tumor model in vivo. Immunohistochemical analyses of tumor samples from NSCLC patients revealed that the expression of pp32/PHAPI correlated with an improved outcome following chemotherapy. These results identify pp32/PHAPI as regulator of the apoptosis response of cancer cells in vitro and in vivo, and as a predictor of survival following chemotherapy for advanced NSCLC.


Microbial background flora in small-scale cheese production facilities does not inhibit growth and surface attachment of Listeria monocytogenes.
The background microbiota of 5 Norwegian small-scale cheese production sites was examined and the effect of the isolated strains on the growth and survival of Listeria monocytogenes was investigated. Samples were taken from the air, food contact surfaces (storage surfaces, cheese molds, and brine) and noncontact surfaces (floor, drains, and doors) and all isolates were identified by sequencing and morphology (mold). A total of 1,314 isolates were identified and found to belong to 55 bacterial genera, 1 species of yeast, and 6 species of mold. Lactococcus spp. (all of which were Lactococcus lactis), Staphylococcus spp., Microbacterium spp., and Psychrobacter sp. were isolated from all 5 sites and Rhodococcus spp. and Chryseobacterium spp. from 4 sites. Thirty-two genera were only found in 1 out of 5 facilities each. Great variations were observed in the microbial background flora both between the 5 producers, and also within the various production sites. The greatest diversity of bacteria was found in drains and on rubber seals of doors. The flora on cheese storage shelves and in salt brines was less varied. A total of 62 bacterial isolates and 1 yeast isolate were tested for antilisterial activity in an overlay assay and a spot-on-lawn assay, but none showed significant inhibitory effects. Listeria monocytogenes was also co-cultured on ceramic tiles with bacteria dominating in the cheese production plants: Lactococcus lactis, Pseudomonas putida, Staphylococcus equorum, Rhodococcus spp., or Psychrobacter spp. None of the tested isolates altered the survival of L. monocytogenes on ceramic tiles. The conclusion of the study was that no common background flora exists in cheese production environments. None of the tested isolates inhibited the growth of L. monocytogenes. Hence, this study does not support the hypothesis that the natural background flora in cheese production environments inhibits the growth or survival of L. monocytogenes.


PCOS is associated with increased CD11c expression and crown-like structures in adipose tissue and increased central abdominal fat depots independent of obesity.
Adipose tissue macrophage (ATM) infiltration is a major pathway for obesity-induced insulin resistance but has not been studied as a mechanism for insulin resistance in PCOS. We tested whether polycystic ovary syndrome (PCOS) is associated with increased ATM infiltration, especially of inflammatory subtype identified by the CD11c marker. We conducted a case-control study at an academic medical center in the United States. Fourteen PCOS and 14 control women of similar age and body mass index (BMI) underwent a gluteal fat biopsy. Markers of ATM, integrins, TNF-α, and adiponectin, were analyzed by quantitative RT-PCR using a standard curve method. Crown-like structures (CLS) were identified by immunohistochemistry. Abdominal magnetic resonance imaging and frequently sampled i.v. glucose tolerance test were performed to assess abdominal fat and insulin sensitivity (SI). Women with PCOS were compared with control women of similar age and BMI for ATM markers, CLS density, adipose tissue expression of inflammatory cytokines and adiponectin, SI, and abdominal fat depots. Women with PCOS had an increase in CD11c expression (P = 0.03), CLS density (P = 0.001), α5 expression (P = 0.009), borderline increase in TNF-α expression (P = 0.08), and a decrease in adiponectin expression (P = 0.02) in gluteal adipose tissue. Visceral (P = 0.009) and sc abdominal fat (P = 0.005) were increased in PCOS. SI was lower in PCOS (P = 0.008). PCOS is associated with an increase in CD11c expression and CLS density and a decrease in adiponectin expression in sc adipose tissue. Additionally, PCOS is associated with higher central abdominal fat depots independent of BMI. These alterations are present among mostly nonobese women and could represent mechanisms for insulin resistance.


Changes in white matter microstructure during adolescence.
Postmortem histological studies have demonstrated that myelination in human brain white matter (WM) continues throughout adolescence and well into adulthood. We used in vivo diffusion-weighted magnetic resonance imaging to test for age-related WM changes in 42 adolescents and 20 young adults. Tract-Based Spatial Statistics (TBSS) analysis of the adolescent data identified widespread age-related increases in fractional anisotropy (FA) that were most significant in clusters including the body of the corpus callosum and right superior corona radiata. These changes were driven by changes in perpendicular, rather than parallel, diffusivity. These WM clusters were used as seeds for probabilistic tractography, allowing us to identify the regions as belonging to callosal, corticospinal, and prefrontal tracts. We also performed voxel-based morphometry-style analysis of conventional T1-weighted images to test for age-related changes in grey matter (GM). We identified a cluster including right middle frontal and precentral gyri that showed an age-related decrease in GM density through adolescence and connected with the tracts showing age-related WM FA increases. The GM density decrease was highly significantly correlated with the WM FA increase in the connected cluster. Age-related changes in FA were much less prominent in the young adult group, but we did find a significant age-related increase in FA in the right superior longitudinal fascicle, suggesting that structural development of this pathway continues into adulthood. Our results suggest that significant microstructural changes in WM continue throughout adolescence and are associated with corresponding age-related changes in cortical GM regions.


Equilibrium and kinetic aspects of the uptake of poly(ethylene oxide) by copolymer microgel particles of N-isopropylacrylamide and acrylic acid.
The use of microgels for controlled uptake and release has been an area of active research for many years. In this work copolymer microgels of N-isopropylacrylamide (NIPAM) and acrylic acid (AAc), containing different concentrations of AAc and also cross-linking monomer, have been prepared and characterized. These microgels are responsive to pH and temperature. As well as monitoring the equilibrium response to changes in these variables, the rates of swelling/de-swelling of the microgel particles, on changing either the pH or the temperature, have also been investigated. It is shown that the rate of de-swelling of the microgel particles containing AAc is much faster than the rate of swelling, on changing the pH appropriately. This is explained in terms of the relative mobilities of the H(+) and Na(+) ions, in and out of the particles. It was observed that the microgels containing AAc, at pH 8, de-swelled relatively slowly on heating to 50 degrees C from 20 degrees C. This is attributed to the resistance to collapse associated with the large increase in counterion concentration inside the microgel particles. The swelling and de-swelling properties of these copolymer microgels have also been investigated in aqueous poly(ethylene oxide) (PEO) solutions, of different MW (2000-300 000). The corresponding absorbed amounts of PEO from solution onto the microgels have also been determined using a depletion method. The results, as a function of AAc content, cross-linker concentration, PEO MW, pH, and temperature, have been rationalized in terms of the ease and depth of penetration of the PEO chains into the various microgel particles and also the H-bonding associations between PEO and either the -COOH of the AAc moeities and/or the H of the amide groups (much weaker). Finally, the adsorption and desorption of the PEO molecules in to and out of the microgel particles have been shown to be extremely slow compared to normal diffusion time scales for polymer adsorption onto rigid surfaces.


Neural circuit formation in the cerebellum is controlled by cell adhesion molecules of the Contactin family.
Cell adhesion molecules of the immunoglobulin superfamily (IgSF CAMs) have been implicated in neural circuit formation in both the peripheral and the central nervous system. Several recent studies highlight a role of the Contactin group of IgSF CAMs in cerebellar development, in particular in the development of granule cells. Granule cells are the most numerous type of neurons in the nervous system and by forming a secondary proliferative zone in the cerebellum they provide an exception to the rule that neuronal precursors proliferate in the ventricular zone. Granule cells express Contactin-2, Contactin-1, and Contactin-6 in a sequential manner. Contactins are required for axon guidance, fasciculation, and synaptogenesis, and thus affect multiple steps in neural circuit formation in the developing cerebellum.


Accurate Classification of Protein Subcellular Localization from High-Throughput Microscopy Images Using Deep Learning.
High-throughput microscopy of many single cells generates high-dimensional data that are far from straightforward to analyze. One important problem is automatically detecting the cellular compartment where a fluorescently-tagged protein resides, a task relatively simple for an experienced human, but difficult to automate on a computer. Here, we train an 11-layer neural network on data from mapping thousands of yeast proteins, achieving per cell localization classification accuracy of 91%, and per protein accuracy of 99% on held-out images. We confirm that low-level network features correspond to basic image characteristics, while deeper layers separate localization classes. Using this network as a feature calculator, we train standard classifiers that assign proteins to previously unseen compartments after observing only a small number of training examples. Our results are the most accurate subcellular localization classifications to date, and demonstrate the usefulness of deep learning for high-throughput microscopy.


Analysis of the mechanism(s) of metaphase I arrest in maturing mouse oocytes.
Fully grown mouse oocytes are normally competent to progress from prophase I to metaphase II without interruption. However, growing mouse oocytes initially become only partially competent to undergo meiotic maturation. Meiotic maturation in these oocytes does not progress beyond metaphase I. In contrast to the oocytes of most strains of mice, most oocytes of strain LT/Sv mice become arrested at metaphase I even when they are fully grown. The initiation of oocyte maturation is correlated with an increase in p34cdc2 kinase activity that continues to rise until metaphase I. The transition into anaphase I is normally correlated with a decrease in p34cdc2 kinase activity. This study demonstrated that metaphase I arrest in both partially competent growing oocytes and fully grown LT/Sv oocytes is correlated with a sustained elevation of p34cdc2 kinase activity. In fact, p34cdc2 activity continued to increase during the time when activity normally decreased. In normally maturing oocytes, some, but not all, of the cyclin B, the regulatory protein associated with p34cdc2, became degraded in oocytes that entered anaphase I. In contrast, the amount of cyclin B present in the metaphase I-arrested oocytes continued to increase at the time when it was being degraded in normal oocytes progressing to metaphase II. These results suggest that the progression of meiosis is arrested at metaphase I in both groups of oocytes because of continued p34cdc2 kinase activity sustained, at least in part, by restricted degradation of cyclin B.(ABSTRACT TRUNCATED AT 250 WORDS)


Concerted regulation of retinal pigment epithelium basement membrane and barrier function by angiocrine factors.
The outer blood-retina barrier is established through the coordinated terminal maturation of the retinal pigment epithelium (RPE), fenestrated choroid endothelial cells (ECs) and Bruch's membrane, a highly organized basement membrane that lies between both cell types. Here we study the contribution of choroid ECs to this process by comparing their gene expression profile before (P5) and after (P30) the critical postnatal period when mice acquire mature visual function. Transcriptome analyses show that expression of extracellular matrix-related genes changes dramatically over this period. Co-culture experiments support the existence of a novel regulatory pathway: ECs secrete factors that remodel RPE basement membrane, and integrin receptors sense these changes triggering Rho GTPase signals that modulate RPE tight junctions and enhance RPE barrier function. We anticipate our results will spawn a search for additional roles of choroid ECs in RPE physiology and disease.


Environmental influence on the evolution of morphological complexity in machines.
Whether, when, how, and why increased complexity evolves in biological populations is a longstanding open question. In this work we combine a recently developed method for evolving virtual organisms with an information-theoretic metric of morphological complexity in order to investigate how the complexity of morphologies, which are evolved for locomotion, varies across different environments. We first demonstrate that selection for locomotion results in the evolution of organisms with morphologies that increase in complexity over evolutionary time beyond what would be expected due to random chance. This provides evidence that the increase in complexity observed is a result of a driven rather than a passive trend. In subsequent experiments we demonstrate that morphologies having greater complexity evolve in complex environments, when compared to a simple environment when a cost of complexity is imposed. This suggests that in some niches, evolution may act to complexify the body plans of organisms while in other niches selection favors simpler body plans.


Targeting the endocannabinoid system for the treatment of cancer--a practical view.
In recent years, considerable interest has been generated by findings that cannabinoids not only have useful palliative effects, but also can affect the viability and invasivity of a variety of different cancer cells. In the present review, the potential of targeting the cannabinoid system for the treatment of cancer is considered from a practical, rather than a mechanistic viewpoint, addressing questions such as whether human tumour cells express CB receptors; whether the potencies of action of cannabinoids in vitro match the potencies expected on the base of receptor theory; what is known about the in vivo effects of cannabinoids and cancer, and how relevant the experiments undertaken are to the clinical situation; and finally, what approaches can be taken to minimise unwanted effects of cannabinoid treatment. It is concluded that cannabinoids (or agents modulating the endogenous cannabinoid system) are an attractive target for drug development in the cancer area, but that more in vivo studies, particularly those investigating the potential of cannabinoids as an addition to current treatment strategies, are needed.


Baseline inflammation is not predictive of periprocedural troponin elevation after elective percutaneous coronary intervention.
High-sensitivity C-reactive protein predicts future cardiovascular events in both healthy individuals and patients with unstable and stable coronary syndromes. Few data are available about the incidence and the relation to inflammation of troponin elevation following percutaneous coronary intervention (PCI), a potential predictor of longterm outcome. We sought to confirm the impact of embolization on long-term outcome and evaluate the ability of baseline inflammation to predict troponin elevation induced by PCI. We prospectively analyzed 200 patients treated by PCI for stable or Braunwald IIA class unstable angina. The patients were recruited between January 1997 and May 1999, and the population was followed during a mean follow-up of 32 months. Major adverse cardiac events (MACEs) were defined as the occurrence of death, myocardial infarction or recurrent angina requiring repeat PCI, or coronary artery bypass grafting. During the follow-up period, 58 MACEs were observed. By multivariate analysis, independent predictors for the occurrence of MACEs were unstable angina and troponin I level after PCI (P < 0.0001 for both). No correlation was found between baseline inflammation and significant troponin I elevation post PCI and by multivariate analysis, no biological variable was a predictor of troponin I elevation post PCI. Baseline inflammation cannot predict onset of minor myonecrosis damage (expressed by troponin elevation) induced by PCI, a significant predictor of long-term outcome in this setting.


Complete Assembly of the Genome of an Acidovorax citrulli Strain Reveals a Naturally Occurring Plasmid in This Species.
Acidovorax citrulli is the causal agent of bacterial fruit blotch (BFB), a serious threat to cucurbit crop production worldwide. Based on genetic and phenotypic properties, A. citrulli strains are divided into two major groups: group I strains have been generally isolated from melon and other non-watermelon cucurbits, while group II strains are closely associated with watermelon. In a previous study, we reported the genome of the group I model strain, M6. At that time, the M6 genome was sequenced by MiSeq Illumina technology, with reads assembled into 139 contigs. Here, we report the assembly of the M6 genome following sequencing with PacBio technology. This approach not only allowed full assembly of the M6 genome, but it also revealed the occurrence of a ∼53 kb plasmid. The M6 plasmid, named pACM6, was further confirmed by plasmid extraction, Southern-blot analysis of restricted fragments and obtention of M6-derivative cured strains. pACM6 occurs at low copy numbers (average of ∼4.1 ± 1.3 chromosome equivalents) in A. citrulli M6 and contains 63 open reading frames (ORFs), most of which (55.6%) encoding hypothetical proteins. The plasmid contains several genes encoding type IV secretion components, and typical plasmid-borne genes involved in plasmid maintenance, replication and transfer. The plasmid also carries an operon encoding homologs of a Fic-VbhA toxin-antitoxin (TA) module. Transcriptome data from A. citrulli M6 revealed that, under the tested conditions, the genes encoding the components of this TA system are among the highest expressed genes in pACM6. Whether this TA module plays a role in pACM6 maintenance is still to be determined. Leaf infiltration and seed transmission assays revealed that, under tested conditions, the loss of pACM6 did not affect the virulence of A. citrulli M6. We also show that pACM6 or similar plasmids are present in several group I strains, but absent in all tested group II strains of A. citrulli.


Molecular cloning and characterisation of GPR74 a novel G-protein coupled receptor closest related to the Y-receptor family.
A novel gene product, GPR74, with homology to the seven transmembrane-domain receptor superfamily, has been cloned. GPR74 has been identified from the expressed sequence tags (EST) database. Subsequent PCR amplification of that sequence and screening of a human heart cDNA library led to the isolation of a 1.7-kb cDNA clone encoding a protein of 408 amino acids. GPR74 shows highest amino acid identity (33%) to the human neuropeptide Y-receptor subtype Y2. The human and mouse genes for GPR74 have been isolated and their exon-intron structures determined. In both species the gene consists of four exons spanning around 20 kb with the exon-intron borders being 100% conserved. Northern analysis of various human tissues reveals highest levels of mRNA expression in brain and heart. In situ hybridisation analysis of rat brain tissue confirms this result and identifies the hippocampus and amygdala nuclei as the brain areas with particular high expression of GPR74 mRNA. Fluorescence in situ hybridisation, PCR analysis on a radiation hybrid panel and interspecific mouse backcross mapping have localised the genes to human chromosome 4q21 and mouse chromosome 5. Expression of the human GPR74 cDNA as a GFP-fusion protein in various cell lines reveals the inability of the recombinant receptor protein to reach the cell surface. This is consistent with the lack of NPY specific binding in these cells and suggests that unknown factors are required for a full functional receptor complex.


Computerized tomography technique for reconstruction of obstructed phase data in shearography.
Shearography is an interferometric method that overcomes several limitations of holography by eliminating the reference beam. It greatly simplifies the optical setup and has much higher tolerance to environmental disturbances. Consequently, the technique has received considerable industrial acceptance, particularly for nondestructive testing. Shearography, however, is generally not applicable to the measurement of an obstructed area, as the area to be measured must be accessible to both illumination and imaging. We present an algorithm based on the principle of tomography that permits the reconstruction of the unavailable phase distribution in an obstructed area from the measured boundary phase distribution. In the process, a set of imaginary rays is projected from many different directions across the area. For each ray, integration of the phase directional derivative along the ray is equal to the phase difference between the boundary points intercepted by the ray. Therefore, a set of linear equations can be established by considering the multiple rays. Each equation expresses the unknown phase derivatives in the obstructed area in terms of the measured boundary phase. Solution of the set of simultaneous equations yields the unknown phase distribution in the blind area. While its applications to shearography are demonstrated, the technique is potentially applicable to all full-field optical measurement techniques such as holography, speckle interferometry, classical interferometry, thermography, moiré, photoelasticity, and speckle correlation techniques.


Do the etiological factors in artificial urinary sphincter reimplantation cases affect success and complications?
The artificial urinary sphincter (AUS) is still one of the best options for incontinence treatment. It may also have an advantage for revision or reimplantation in the management of complications. In this study we aimed to discuss the etiological factors for AUS reimplantation and effects of these etiological factors on success rates, patient satisfaction rates, time to reimplantation surgery, and complications Data from 30 patients for whom AUS reimplantation was performed were analyzed retrospectively. Incontinence due to fluid loss from the cuff or reservoir balloon, inability of the cuff to adequately compress the urethra, and devices that were thought to have completed their lifespans were defined as mechanical reasons while incontinence caused by conditions such as cuff erosion and infection were defined as nonmechanical reasons. Patients who went through reimplantation due to mechanical and nonmechanical causes were included in Group 1 and Group 2, respectively. Success rates, patient satisfaction rates, time between the implantation of the first and second AUS, and complications were compared between the groups. The mean follow-up period was 79 (3–308) months for patients who went through primary AUS implantation due to postprostatectomy incontinence. Our success rates were found as 75% and 66% in Group 1 and Group 2, respectively. The differences between the groups in terms of success and patient satisfaction rates were not statistically significant, while the time to reimplantation was longer in Group 1 and statistically significant. Reasons for AUS reimplantation may affect the success and patient satisfaction rates. Our success rates of AUS performed for nonmechanical reasons were slightly lower, but not statistically significantly so. AUS reimplantation may take a longer time if mechanical failure is detected.


Differential modulation of the baroreceptor reflex by brain and plasma vasopressin.
Plasma vasopressin sensitizes the baroreceptor reflex, whereas vasopressin given into the cerebral ventricle overrides the baroreceptor reflex by means of sympathetic stimulation. To test the hypothesis that arginine vasopressin stimulates two different receptor subtypes (V1 and V2) in the central nervous system, we measured the baroreceptor reflex (change in pulse interval vs change in blood pressure) after administering methoxamine (10-300 micrograms/kg i.v.) in conscious rats. Animals were pretreated either with a V1 vasopressin receptor antagonist administered intravenously or intracerebroventricularly, or with a V2 receptor antagonist administered intravenously. The central V1 antagonist caused sensitization of the baroreceptor reflex, whereas the intravenous V2 antagonist attenuated it. The intravenous V1 vasopressin antagonist had no effect on baroreceptor reflex sensitivity. When the experiments were repeated in rats with hereditary diabetes insipidus, neither antagonist influenced the baroreceptor reflex. Volume expansion lowered circulating vasopressin levels and also attenuated the baroreceptor reflex--effects similar to those observed with the intravenous V2 antagonist. We conclude that vasopressin sensitizes the baroreceptor reflex through V2 receptors accessible from the blood and inhibits the reflex through V1 receptors in the brain that cannot be reached from the blood. These observations suggest a direct interaction between hormonal and neuronal vasopressin in cardiovascular control.


Second-hand smoke, cotinine levels, and risk of circulatory mortality in a large cohort study of never-smokers.
Exposure to second-hand smoke has been shown to be associated with increased cardiovascular mortality in several, but not all, epidemiologic studies. Our aim was to investigate the risk of circulatory death associated with exposure to second-hand smoke in never-smokers in a very large prospective study, the European Prospective Investigation into Cancer and Nutrition. A secondary aim was to use cotinine levels for cross-validating self-reported second-hand smoke exposure. Cox proportional hazard models were used to investigate the risk of death due to circulatory causes associated with second-hand smoke exposure in 135,233 never-smokers. Exposure to second-hand smoke was assessed through a questionnaire at enrollment and then validated against plasma cotinine measurements in a subsample. Study participants who reported second-hand smoke exposure at home had higher cotinine levels (median plasma cotinine concentration in exposed = 0.82 microg/L; in those unexposed 0.02 microg/L). Second-hand smoke exposure at home was associated with an increased risk of dying from cardiovascular diseases (hazard ratio [HR] = 1.38 [95% confidence interval = 1.01-1.90]), all circulatory diseases (1.28 [0.98-1.69]), and coronary heart disease (1.31 [0.83-2.08]) after adjustment for age, sex, education, physical activity, and body mass index. Dose-response relationships were observed between exposure to second-hand smoke at home and risk of circulatory death (HR per each additional hour/d = 1.25 [1.04-1.50]). Having a partner who smokes more than 30 cigarettes per day considerably increased the risk of a circulatory death (2.94 [1.11-7.78]). Second-hand smoke exposure at home was not associated with total mortality (1.03 [0.93-1.13]). Exposure to second-hand smoke at home (as confirmed by plasma cotinine levels) increases the risk of cardiovascular mortality.


Dr. Probe: A software for high-resolution STEM image simulation.
The Dr. Probe software for multislice simulations of STEM images is introduced, and reference is given of the applied methods. Major program features available with the graphical user interface version are demonstrated by means of a few examples for bright-field and dark-field STEM imaging as well as simulations of diffraction patterns. The numerical procedure applied for the simulation of thermal-diffuse scattering by the frozen-lattice approach is described in detail. Intensity variations occurring in simulations with atomic-column resolution due to frozen-lattice variations are discussed in the context of atom counting. It is found that a significant averaging over many lattice configurations with different random atomic displacements is required to prevent atom-counting bias from simulations. A strategy is developed for the assessment of the amount of required averaging based on the estimated signal variance and the expected signal gain per atom in a column.


Isolation of plasmid deoxyribonucleic acid from Pseudomonas putida.
Conditions suitable for reproducible recovery of covalently closed circular deoxyribonucleic acid from strains of Pseudomonas putida containing degradative plasmids (CAM, SAL, OCT, etc.) have been defined. These degradative plasmids could not be isolated by the usual procedure, whereas RP1, an R factor of the P group, present in the isogenic strain of P. putida, was isolated equally well by either the usual procedure or the modified procedure. Characterization by electron microscopy of RP1 deoxyribonucleic acid confirmed the molecular weight (about 40 X 10(6)) previously determined by sucrose gradient centrifugation.


The Filament Editor: an interactive software environment for visualization, proof-editing and analysis of 3D neuron morphology.
Neuroanatomical analysis, such as classification of cell types, depends on reliable reconstruction of large numbers of complete 3D dendrite and axon morphologies. At present, the majority of neuron reconstructions are obtained from preparations in a single tissue slice in vitro, thus suffering from cut off dendrites and, more dramatically, cut off axons. In general, axons can innervate volumes of several cubic millimeters and may reach path lengths of tens of centimeters. Thus, their complete reconstruction requires in vivo labeling, histological sectioning and imaging of large fields of view. Unfortunately, anisotropic background conditions across such large tissue volumes, as well as faintly labeled thin neurites, result in incomplete or erroneous automated tracings and even lead experts to make annotation errors during manual reconstructions. Consequently, tracing reliability renders the major bottleneck for reconstructing complete 3D neuron morphologies. Here, we present a novel set of tools, integrated into a software environment named 'Filament Editor', for creating reliable neuron tracings from sparsely labeled in vivo datasets. The Filament Editor allows for simultaneous visualization of complex neuronal tracings and image data in a 3D viewer, proof-editing of neuronal tracings, alignment and interconnection across sections, and morphometric analysis in relation to 3D anatomical reference structures. We illustrate the functionality of the Filament Editor on the example of in vivo labeled axons and demonstrate that for the exemplary dataset the final tracing results after proof-editing are independent of the expertise of the human operator.


Nitrous Oxide Emissions Increase Exponentially When Optimum Nitrogen Fertilizer Rates Are Exceeded in the North China Plain.
The IPCC assume a linear relationship between nitrogen (N) application rate and nitrous oxide (N2O) emissions in inventory reporting, however, a growing number of studies show a nonlinear relationship under specific soil-climatic conditions. In the North China plain, a global hotspot of N2O emissions, covering a land as large as Germany, the correlation between N rate and N2O emissions remains unclear. We have therefore specifically investigated the N2O response to N applications by conducting field experiments with five N rates, and high-frequency measurements of N2O emissions across contrasting climatic years. Our results showed that cumulative and yield-scaled N2O emissions both increased exponentially as N applications were raised above the optimum rate in maize ( Zea mays L.). In wheat ( Triticum aestivum L.) there was a corresponding quadratic increase in N2O emissions with the magnitude of the response in 2012-2013 distinctly larger than that in 2013-2014 owing to the effects of extreme snowfall. Existing empirical models (including the IPCC approach) of the N2O response to N rate have overestimated N2O emissions in the North China plain, even at high N rates. Our study therefore provides a new and robust analysis of the effects of fertilizer rate and climatic conditions on N2O emissions.


Patient selection critical for calcineurin inhibitor withdrawal in pediatric kidney transplantation.
CNI withdrawal may be employed as a "rescue" strategy for patients with established renal allograft injury and/or declining allograft function, with the aim at eliminating CNI-associated nephrotoxic effects. This analysis reviews outcomes in a pediatric population and identifies risk factors for adverse events post-CNI withdrawal. We performed a retrospective analysis of 17 pediatric renal transplants who underwent CNI withdrawal, with conversion to sirolimus and MMF. Mean CrCl decreased from 64.3 +/- 22 to 59.38 +/- 28.6 mL/min/1.73 m(2) (p = 0.04) at six months and 57.46 +/- 31.1 mL/min/1.73 m(2) (p = 0.02) at 12 months post-withdrawal. Forty-one percent of patients experienced AR. Increased risk for AR was associated with prior AR history, lower sirolimus trough levels, and lower CNIT biopsy scores. Graft loss (24%) was associated with worse CrCl, proteinuria, and histologic chronicity. Proteinuria (spot protein/creatinine ratio) increased from 0.75 +/- 1.0 to 1.71 +/- 2.0 (p = 0.03), unrelated to de novo sirolimus use. Four patients returned to CNI-based immunosuppression due to AR (n = 3) and gastrointestinal side effects (n = 1). Careful selection of pediatric candidates for CNI withdrawal is recommended. Worsening graft function and graft loss may be minimized by selecting patients with high CNIT scores and low biopsy chronicity and excluding patients with prior AR history.


Exploring men's pregnancy-planning behaviour and fertility knowledge:a survey among fathers in Sweden.
Research about pregnancy-planning behaviour mostly focuses on women, even though pregnancy planning usually also concerns men. The purpose of this study was to investigate how men plan for family, and to measure their fertility knowledge after having become fathers. Data were collected in 2014 as part of a Swedish longitudinal pregnancy-planning study. Men were recruited through their female partner one year after childbirth. Participants were asked to fill out a questionnaire about pregnancy planning, lifestyles, and fertility. Of the 796 participants, 646 (81%) stated that the pregnancy had been very or fairly planned, and 17% (n = 128) had made a lifestyle adjustment before pregnancy to improve health and fertility. The most common adjustments were to reduce/quit the consumption of alcohol, cigarettes, or snuff, and to exercise more. First-time fathers and those who had used assisted reproductive technology to become pregnant were more likely to have made an adjustment. Fertility knowledge varied greatly. Men with university education had better fertility knowledge than men without university education. Our findings indicate that there is variation in how men plan and prepare for pregnancy. Most men did not adjust their lifestyle to improve health and fertility, while some made several changes. Both pregnancy-planning behaviour and fertility knowledge seem to be related to level of education and mode of conception. To gain deeper understanding of behaviour and underlying factors, more research is needed.


Improving the Health of Older Prisoners: Nutrition and Exercise in Correctional Institutions.
This qualitative article presents and compares the views of older prisoners and expert stakeholders on the topic of nutrition and exercise. The study highlights measures for improving the health of older prisoners. Older prisoners report the need to improve quality of meals provided in prison. They note that prison food is of poor quality and not adapted to their needs. With regard to exercise, they point out the lack of proper opportunities to engage in exercise and sports, and describe several factors that make physical activities either unsuitable or unfeasible. Expert stakeholders see prison as an opportunity to improve the health of those convicted of crimes. In light of the increasing number of older prisoners, age-appropriate nutritional and exercise interventions should take priority in prisons.


Selective inhibition of renal thromboxane biosynthesis increased sodium excretion rate in normal and saline-loaded rats.
The excretion rates of renal thromboxane B2 (TXB2) and 6-ketoPGF1 alpha, the stable chemical metabolites of thromboxane A2 (TXA2) and prostaglandin I2 (PGI2) respectively, PGE2 and sodium were determined in normal and saline-loaded rats treated with the thromboxane synthetase inhibitor imidazole. In normal rats the administration of imidazole in doses which did not affect renal 6-keto-PGF1 alpha and PGE2 excretion but selectively inhibited renal TXB2 excretion, significantly increased the sodium excretion rate. Volume expansion with saline increased renal PGE2 and 6-ketoPGF1 alpha excretion but did not alter renal TXB2 excretion. The increase in renal prostaglandin excretion was accompanied by an increased sodium excretion rate. The administration of imidazole to saline-loaded animals also decreased renal TXB2 excretion but did not alter the increased excretion of renal PGE2 and 6-ketoPGF1 alpha. This reduction in renal thromboxane biosynthesis by imidazole further increased the sodium excretion rate. We suggest that TXA2 is a potent antinatriuretic factor as well as the most potent vasoconstrictor agent known.


Refractory myasthenia gravis: Characteristics of a portuguese cohort.
Some myasthenia gravis (MG) patients are refractory to conventional treatments. To describe the clinical features of refractory MG (RMG) and explore the association with human leukocyte antigen HLA-DRB1 alleles, a cohort study of 114 consecutive MG patients was performed. Patients were classified as RMG based on predefined criteria. Twenty-two patients were found to have RMG (19.3%). There were no differences between non-RMG and RMG patients with respect to sex, age of onset, abnormal 3-Hz repetitive nerve stimulation, anti-acetylcholine receptor antibody positivity, thymectomy, thymoma or thymic hyperplasia, and polyautoimmunity. HLA-DRB1*03 was more frequent in the non-RMG vs. control population (P = 3 × 10-6 ). The HLA-DRB1*13 allele was less frequent in non-RMG patients compared with controls (P = 0.002), and less frequent in the non-RMG group compared with the RMG group (P = 0.003). HLA-DRB1*03 was more common in non-RMG, and the HLA-DRB1*13 allele appeared to have a protective role, as reported previously in other autoimmune disorders. Muscle Nerve 60: 188-191, 2019.


Scheffersomyces cryptocercus: a new xylose-fermenting yeast associated with the gut of wood roaches and new combinations in the Sugiyamaella yeast clade.
The gut of wood-feeding insects is a microhabitat for a specialized community of microbes, including bacteria and several groups of eukaryotes such as nematodes, parabasalids and fungi. The characterization of gut yeast communities from a variety of insects has shown that certain yeasts often are associated with the insects. The gut of wood-feeding insects is rich in ascomycete yeasts and in particular xylose-fermenting (X-F) and assimilating yeasts have been consistently present in the gut of lignicolous insects. The objective of this study was the characterization of the yeast flora from the gut of the wood roach Cryptocercus sp. (Blattodea: Cryptocercidae). Five wood roaches were collected along the Appalachian Trail near the border between Tennessee and North Carolina, USA. We isolated 18 yeast strains from the wood roaches identified as Sugiyamaella paludigena and Sugiyamaella lignohabitans, xylose-assimilating yeasts, and Scheffersomyces cryptocercus (NRRL Y-48824(T) = CBS 12658) a new species of X-F yeast. The presence of X-F and certain non X-F yeasts in the gut of the subsocial wood roach Cryptocercus sp. extends the previous findings of associations between certain ascomycete yeasts and lignicolous insects. New combinations were made for 13 asexual members of the Sugiyamaella clade.


The frequency of surgical maneuvers during open rhinoplasty.
The improved exposure afforded by the external rhinoplasty approach has allowed for more precise surgical maneuvers and makes more consistent results possible. This study examines the frequency and array of surgical maneuvers during primary aesthetic rhinoplasty in a single surgeon's practice. A retrospective review of 100 consecutive primary external cosmetic rhinoplasty patients with a minimum follow-up of approximately 1 year was conducted. Surgical maneuvers were reviewed and tabulated. Special attention was devoted to the number of cartilage grafts and suturing techniques used. Operative times and incidence of revision surgery were investigated. The mean duration of surgery was 1 hour 50 minutes. The majority of rhinoplasties involved concomitant septal surgery for deviation and airway occlusion or cartilage harvesting. Osteotomies were performed on over 95 percent of patients. The dorsal hump was removed on 84 percent of the patients. The most common grafts used included alar rim grafts in 88 percent, subdomal grafts in 77 percent, and spreader grafts in 74 percent. Sutures used to contour the cartilage included transdomal sutures in 62 percent, followed by footplate approximation sutures in 32 percent. Seventy-seven patients had interdomal sutures placed as a component of subdomal grafting. The senior surgeon has found that common incorporation of certain maneuvers offers more consistent, aesthetically pleasing, and superior functional outcomes. Careful review of this article may guide the reader to consider the more frequent use of some maneuvers or reduction of other maneuvers to reflect that of a streamlined and higher-volume rhinoplasty practice.


The man who mistook his neuropsychologist for a popstar: when configural processing fails in acquired prosopagnosia.
We report the case of an individual with acquired prosopagnosia who experiences extreme difficulties in recognizing familiar faces in everyday life despite excellent object recognition skills. Formal testing indicates that he is also severely impaired at remembering pre-experimentally unfamiliar faces and that he takes an extremely long time to identify famous faces and to match unfamiliar faces. Nevertheless, he performs as accurately and quickly as controls at identifying inverted familiar and unfamiliar faces and can recognize famous faces from their external features. He also performs as accurately as controls at recognizing famous faces when fracturing conceals the configural information in the face. He shows evidence of impaired global processing but normal local processing of Navon figures. This case appears to reflect the clearest example yet of an acquired prosopagnosic patient whose familiar face recognition deficit is caused by a severe configural processing deficit in the absence of any problems in featural processing. These preserved featural skills together with apparently intact visual imagery for faces allow him to identify a surprisingly large number of famous faces when unlimited time is available. The theoretical implications of this pattern of performance for understanding the nature of acquired prosopagnosia are discussed.


Role of specific quorum-sensing signals in the regulation of exopolysaccharide II production within Sinorhizobium meliloti spreading colonies.
Quorum sensing (QS) in Sinorhizobium meliloti involves at least half a dozen different N-acyl homoserine lactone (AHL) signals. These signals are produced by SinI, the sole AHL synthase in S. meliloti Rm8530. The sinI gene is regulated by two LuxR-type transcriptional regulators, SinR and ExpR. Mutations in sinI, sinR and expR abolish the production of exopolysaccharide II (EPS II). This study investigated a new type of coordinated surface spreading of Rm8530 that can be categorized as swarming. Motility assays on semi-solid surfaces revealed that both flagella and EPS II are required for this type of motility. The production of EPS II depends on AHLs produced by SinI. Of these AHLs, only C(16:1)- and 3-oxo-C(16:1)-homoserine lactones (HSLs) stimulated swarming in an ExpR-dependent manner. These two AHLs induced the strongest response in the wggR reporter fusions. WggR is a positive regulator of the EPS II biosynthesis gene expression. The levels of the wggR activation correlated with the extent of swarming. Furthermore, swarming of S. meliloti required the presence of the high molecular weight (HMW) fraction of EPS II. Within swarming colonies, a recombinase-based RIVET reporter in the wggR gene was resolved in 30% of the cells, indicating an enhanced regulation of EPS II production in the subpopulation of cells, which was sufficient to support swarming of the entire colony. Swarming behavior of S. meliloti Rm8530 on semi-solid surfaces is found to be dependent on the functional QS regulatory cascades. Even though multiple AHL signals are produced by the bacterium, only two AHLs species, C(16:1)- and 3-oxo-C(16:1)-HSLs, affected swarming by up-regulating the expression of wggR. While EPS II is produced by Rm8530 as high and low molecular weight fractions, only the HMW EPS II facilitated initial stages of swarming, thus, suggesting a function for this polymer.


Conducting research on sensitive topics with adolescents: ethical and developmental considerations.
Promotion of adolescent health requires well-designed scientific studies that determine the prevalence of the problem of interest, identify risk and resilience factors, and evaluate methods for prevention and intervention. Many adolescent-related health problems are typically considered sensitive by society (e.g., sexual and substance use behaviors), thus further complicating the research process. Using the principles of the Belmont Report as its framework, this paper draws on developmental theories to discuss ethical issues specific to the conduct of research with adolescents. Our ability to use developmentally sensitive research practices will be enhanced by further understanding of issues associated with risk and benefit assessment by the adolescent, their parents, and institutional review boards, and by delineating ways to ensure that adolescent participants are adequately protected and have a developmentally affirming experience.


The "work" of women when considering and using interventions to reduce mother-to-child transmission (MTCT) of HIV.
This paper explores HIV-infected women's experiences of considering and using recommended interventions during pregnancy and postpartum to reduce mother-to-child transmission of HIV. Data were collected from 45 HIV-infected women aged 18-44 years living in Melbourne, Australia. A semi-structured interview was used to collect qualitative information on women's reproductive experience and intentions. The 15 women who had their children after their HIV diagnosis engaged in significant work including surveillance and safety work to minimise stigma and infection, information work to inform decisions and actions, accounting work to calculate risk and benefit, hope and worry work concerning a child's infection status and impact of interventions, work to redefine an acceptable maternal identity, work to prepare an alternative story to counter the disclosure effect of the intervention and emotional work to reconcile guilt when considering these interventions. This study provides a framework to help clinicians understand the real and on-going "work" that women engage in when they are considering interventions recommended by their physicians to reduce transmission of HIV. Even in circumstances where access to and acceptance of interventions are high, women continue to engage in this work even after they have a made a decision about a particular intervention.


Online yoga in myeloproliferative neoplasm patients: results of a randomized pilot trial to inform future research.
Myeloproliferative neoplasm (MPN) patients suffer from significant symptoms, inflammation and reduced quality of life. Yoga improves these outcomes in other cancers, but this hasn't been demonstrated in MPNs. The purpose of this study was to: (1) explore the limited efficacy (does the program show promise of success) of a 12-week online yoga intervention among MPN patients on symptom burden and quality of life and (2) determine feasibility (practicality: to what extent a measure can be carried out) of remotely collecting inflammatory biomarkers. Patients were recruited nationally and randomized to online yoga (60 min/week of yoga) or wait-list control (asked to maintain normal activity). Weekly yoga minutes were collected with Clicky (online web analytics tool) and self-report. Those in online yoga completed a blood draw at baseline and week 12 to assess inflammation (interleukin-6, tumor necrosis factor-alpha [TNF-α]). All participants completed questionnaires assessing depression, anxiety, fatigue, pain, sleep disturbance, sexual function, total symptom burden, global health, and quality of life at baseline, week seven, 12, and 16. Change from baseline at each time point was computed by group and effect sizes were calculated. Pre-post intervention change in inflammation for the yoga group was compared by t-test. Sixty-two MPN patients enrolled and 48 completed the intervention (online yoga = 27; control group = 21). Yoga participation averaged 40.8 min/week via Clicky and 56.1 min/week via self-report. Small/moderate effect sizes were generated from the yoga intervention for sleep disturbance (d = - 0.26 to - 0.61), pain intensity (d = - 0.34 to - 0.51), anxiety (d = - 0.27 to - 0.37), and depression (d = - 0.53 to - 0.78). A total of 92.6 and 70.4% of online yoga participants completed the blood draw at baseline and week 12, respectively, and there was a decrease in TNF-α from baseline to week 12 (- 1.3 ± 1.5 pg/ml). Online yoga demonstrated small effects on sleep, pain, and anxiety as well as a moderate effect on depression. Remote blood draw procedures are feasible and the effect size of the intervention on TNF-α was large. Future fully powered randomized controlled trials are needed to test for efficacy. This trial was retrospectively registered with clinicaltrials.gov (ID: NCT03503838 ) on 4/19/2018.


Stage 1 treatment of pediatric overweight and obesity: a pilot and feasibility randomized controlled trial.
Staged clinical treatment of pediatric obesity is recommended, but untested. Understanding the lowest intensity stage's effectiveness is necessary for future research. This was a randomized controlled trial of children ages 4 to <9 years. Participants were recruited after routine evaluations at a primary care pediatric clinic revealed a BMI ≥85th percentile. The intervention was patterned after the "Prevention plus, Stage 1" treatment recommended by an expert committee. Groups were compared for changes, over a 3-month time period, in BMI z-score and parental reports of behavioral issues related to childhood obesity using intent-to-treat (ITT) analysis. Seventy-two (30% of eligible) children were enrolled and 64 were remeasured at 3-month follow-up. ITT analysis revealed that both groups improved mean BMI z-score [adjusted change -0.07, control, and -0.04, intervention; 95% confidence interval (CI) of difference=-0.14-0.20]. Over half of the children in each group improved their BMI z-score (adjusted proportion decreasing=55% in control vs. 72% in intervention; 95% CI of difference=-0.07-0.42). The intervention group improved comparatively to the control group on numerous behavioral indicators. Implementation of the lowest intensity stage of current recommendations is feasible and possibly of benefit toward lifestyle changes. Results of this study can be used by future clinical researchers designing protocols to test the full multi-staged approach for the treatment of pediatric overweight and obesity in primary care clinical settings.


Evaluation of small (</= 3 cm) renal masses with MDCT: benefits of thin overlapping reconstructions.
Our purpose was to determine whether thin overlapping reconstructions using MDCT improve the detection and characterization of small renal masses. Thirty-seven patients were scanned with MDCT using 2.5-mm collimation. Nephrographic phase data were reconstructed in two ways: a standard protocol (5-mm section thickness, no overlap) and an experimental protocol (3-mm section thickness, 50% overlap). Masses were detected and classified into three groups: group 1, measuring less than 20 H on both protocols (classified as cysts on both); group 2, measuring 20 H or greater on standard protocol and less than 20 H on experimental protocol (classified as cysts using experimental protocol only); and group 3, measuring 20 H or greater on both protocols (not classified as cysts using either protocol). Masses 10 mm or larger in group 3 were evaluated further for enhancement. Statistically significant differences between protocols were assessed using an analysis of counts and proportions. Of 175 detected lesions, 29 (17%) were detected only with the experimental protocol; all but one were smaller than 5 mm. Using the experimental protocol, of 45 masses between 5 and 10 mm, the number of masses that could be characterized as cysts increased from 13 (29%) to 38 (84%). The overall number of indeterminate lesions was reduced from 101 (69%) of 146 lesions detected with the standard protocol to 86 (53%) of 161 lesions detected with the experimental protocol. Using MDCT and thin overlapping reconstructions, renal cysts as small as 5 mm can be diagnosed with more confidence than is possible with standard reconstructions, and the overall number of indeterminate renal masses is reduced.


Prognosis of ventricular arrhythmias in relation to sudden cardiac death: therapeutic implications.
The hypothesis that ventricular arrhythmias represent an independent predictor of sudden cardiac death was examined by analyzing the published data. The frequency and complexity of ventricular arrhythmias increase progressively both with age and severity of heart disease, but no age- or disease-related norms have been established for clinical guidance. Simple and complex arrhythmias, including short runs of ventricular tachycardia, do not increase risk of sudden cardiac death in subjects without heart disease or with heart disease and normal myocardial function. Progression of nonsustained into sustained ventricular tachycardia in such individuals is rare. Simple and complex ventricular arrhythmias are not strong independent predictors of sudden death in survivors of myocardial infarction. In these, the overall incidence of sudden cardiac death averages 3.5 to 5% during the first year, but is about 15 to 20% per year in patients with severely impaired ventricular function. The results of this survey suggest that in patients with well preserved ventricular function, prophylactic use of antiarrhythmic drugs is not indicated, and that treatment of asymptomatic or mildly symptomatic ventricular arrhythmias is not likely to reduce the incidence of sudden cardiac death.


Watercress supplementation in diet reduces lymphocyte DNA damage and alters blood antioxidant status in healthy adults.
Cruciferous vegetable (CV) consumption is associated with a reduced risk of several cancers in epidemiologic studies. The aim of this study was to determine the effects of watercress (a CV) supplementation on biomarkers related to cancer risk in healthy adults. A single-blind, randomized, crossover study was conducted in 30 men and 30 women (30 smokers and 30 nonsmokers) with a mean age of 33 y (range: 19-55 y). The subjects were fed 85 g raw watercress daily for 8 wk in addition to their habitual diet. The effect of supplementation was measured on a range of endpoints, including DNA damage in lymphocytes (with the comet assay), activity of detoxifying enzymes (glutathione peroxidase and superoxide dismutase) in erythrocytes, plasma antioxidants (retinol, ascorbic acid, alpha-tocopherol, lutein, and beta-carotene), plasma total antioxidant status with the use of the ferric reducing ability of plasma assay, and plasma lipid profile. Watercress supplementation (active compared with control phase) was associated with reductions in basal DNA damage (by 17%; P = 0.03), in basal plus oxidative purine DNA damage (by 23.9%; P = 0.002), and in basal DNA damage in response to ex vivo hydrogen peroxide challenge (by 9.4%; P = 0.07). Beneficial changes seen after watercress intervention were greater and more significant in smokers than in nonsmokers. Plasma lutein and beta-carotene increased significantly by 100% and 33% (P < 0.001), respectively, after watercress supplementation. The results support the theory that consumption of watercress can be linked to a reduced risk of cancer via decreased damage to DNA and possible modulation of antioxidant status by increasing carotenoid concentrations.


Isolation of strawberry anthocyanin-rich fractions and their mechanisms of action against murine breast cancer cell lines.
The aim of this study was the evaluation of the effects of strawberry anthocyanin extract treatment on two in vitro models of murine breast cancer cell lines, in an attempt to detect a specific pathway (AMP-activated protein kinase or AMPK) through which strawberries exert their anticancer activity. The anticancer activity of purified anthocyanin extracts from an Alba cultivar on two murine cancer cell lines, N202/1A (with high levels of the HER2/neu oncogene) and N202/1E (with low levels of the HER2/neu oncogene), was evaluated after 48 and 72 h of treatment. The cell viability and apoptosis, intracellular ROS rates, and cell oxidative damage were assessed. Western blot assays were performed to analyze the expression of several proteins related to apoptosis, autophagy, metastasis, the oxidative status, mitochondrial functionality, and the AMPK pathway. This study demonstrated that the anthocyanin extract of Alba strawberry shows an antiproliferative effect on cancer cells, through the induction of apoptosis and oxidative stress, by stimulating different molecular pathways. This study is one of the first studies that have tried to deepen the understanding of a candidate pathway for the explanation of the effects of strawberry on cancer cells. A relationship between the AMPK pathway and the anticancer effects of strawberries was demonstrated.


HIV-infected men who have sex with men who identify themselves as belonging to subcultures are at increased risk for hepatitis C infection.
Hepatitis C virus (HCV) emerged as sexually transmitted infection among HIV-infected men who have sex with men (MSM). We studied whether HCV circulated in identifiable high-risk MSM subcultures and performed phylogenetic analysis. HIV-infected MSM were recruited at the sexually transmitted infections (STI) outpatient clinic and a university HIV clinic in Amsterdam, the Netherlands, 2008-2009. Participants completed a detailed questionnaire and were tested for HCV antibodies and RNA, with NS5B regions sequenced for analysis of clusters. Among 786 participants, the median age was 43 (IQR 37-48) years, and 93 (11.8%) were HCV-positive. Seropositivity was associated with belonging to subcultures identified as leather (aOR 2.60; 95% CI 1.56-4.33), rubber/lycra (aOR 2.15; 95% CI 1.10-4.21), or jeans (aOR 2.23; 95% CI 1.41-3.54). The two largest HCV-RNA monophyletic clusters were compared; MSM in cluster I (genotype 1a, n = 13) reported more partners (P = 0.037) than MSM in cluster II (genotype 4d, n = 14), but demographics, subculture characteristics and other risk behaviors did not differ significantly between the two clusters. HCV infection is associated with identifiable groups of leather/rubber/lycra/jeans subcultures among HIV-infected MSM. Separate epidemiological HCV transmission networks were not revealed. Active HCV screening and treatment within specific subcultures may reduce HCV spread among all MSM.


Tumor stroma-derived Wnt5a induces differentiation of basal cell carcinoma of Ptch-mutant mice via CaMKII.
Basal cell carcinoma (BCC) is the most common skin tumor in humans. Although BCCs rarely metastasize, they can cause significant morbidity due to local aggressiveness. Approximately 20% of BCCs show signs of spontaneous regression. The understanding of molecular events mediating spontaneous regression has the potential to reduce morbidity of BCC and, potentially, other tumors, if translated into tumor therapies. We show that BCCs induced in conditional Ptch(flox/flox)ERT2(+/-) knockout mice regress with time and show a more differentiated phenotype. Differentiation is accompanied by Wnt5a expression in the tumor stroma, which is first detectable at the fully developed tumor stage. Coculture experiments revealed that Wnt5a is upregulated in tumor-adjacent macrophages by soluble signals derived from BCC cells. In turn, Wnt5a induces the expression of the differentiation marker K10 in tumor cells, which is mediated by Wnt/Ca(2+) signaling in a CaMKII-dependent manner. These data support a role of stromal Wnt5a in BCC differentiation and regression, which may have important implications for development of new treatment strategies for this tumor. Taken together, our results establish BCC as an easily accessible model of tumor regression. The regression of BCC despite sustained Hedgehog signaling activity seems to be mediated by tumor-stromal interactions via Wnt5a signaling.


Effect of diabetes on the glycosaminoglycan component of the human glomerular basement membrane.
The glycosaminoglycan (heparan sulfate) component of glomerular basement membranes from human kidneys of diabetic and nondiabetic subjects has been quantitated after isolation from protease digests of the membranes on DEAE-cellulose microcolumns. A significant decrease (P less than 0.005) in the glycosaminoglycan content of diabetic membranes was observed. Heparan sulfate was identified as the predominant glycosaminoglycan in both diabetic and control subjects and the extent of its sulfation appeared to be similar. The reduced level of glycosaminoglycan in the diabetic glomerular basement membrane was accompanied by a significant elevation of hexoses, which are primarily associated with the collagen component, suggesting that a redistribution of basement membrane macromolecules occurs in the diabetic state. Since heparan sulfate has been implicated as a major component of the glomerular anionic filtration barrier, its decreased content in diabetic basement membranes may contribute to the proteinuria observed in this disease.


The time course of development of alcohol-related problems in men and women.
A predictable sequence of alcohol-related problems has been hypothesized to be applicable to the clinical course of alcoholism. However, few recent data are available on this question. The age of first occurrence of 44 alcohol-related life experiences was determined for 478 DSM-III-R defined alcohol-dependent (alcoholic) individuals (317 men and 161 women), and for 444 drinking but not alcohol-dependent subjects (183 men and 261 women). Data were gathered through personal interviews with alcohol-dependent subjects and their relatives using a structured psychiatric interview (SSAGA). A high level of similarity (Spearman's rho = .81, p = .0004) was found for the retrospective reports of the order of appearance of alcohol-related problems between the present sample and an analysis of 636 alcoholic male inpatients who participated in a prior study. Within the present group of 478 alcoholics, the order of appearance of alcohol-related problems was similar for men and women (rho = .84, p < .0001), and the time course of development of problems was similar for treated and untreated alcoholic subgroups (rho = .86, p < .001). Analyses of 19 alcohol-related life experiences in 444 drinking but not alcohol-dependent individuals indicated an overall rank order for occurrence of problems similar to those observed for alcohol-dependent individuals (rho = .76, p < .001). These data corroborate the high level of predictability regarding the order of occurrence of major alcohol-related life problems among alcohol-dependent men and women, extending the previous findings to women with alcohol dependence and to alcoholics who have never received inpatient treatment.


Online engagement with 2020 election misinformation and turnout in the 2021 Georgia runoff election.
Following the 2020 general election, Republican elected officials, including then-President Donald Trump, promoted conspiracy theories claiming that Joe Biden's close victory in Georgia was fraudulent. Such conspiratorial claims could implicate participation in the Georgia Senate runoff election in different ways-signaling that voting doesn't matter, distracting from ongoing campaigns, stoking political anger at out-partisans, or providing rationalizations for (lack of) enthusiasm for voting during a transfer of power. Here, we evaluate the possibility of any on-average relationship with turnout by combining behavioral measures of engagement with election conspiracies online and administrative data on voter turnout for 40,000 Twitter users registered to vote in Georgia. We find small, limited associations. Liking or sharing messages opposed to conspiracy theories was associated with higher turnout than expected in the runoff election, and those who liked or shared tweets promoting fraud-related conspiracy theories were slightly less likely to vote.


Identification of pathological RA endotypes using blood-based biomarkers reflecting tissue metabolism. A retrospective and explorative analysis of two phase III RA studies.
There is an increasing demand for accurate endotyping of patients according to their pathogenesis to allow more targeted treatment. We explore a combination of blood-based joint tissue metabolites (neoepitopes) to enable patient clustering through distinct disease profiles. We analysed data from two RA studies (LITHE (N = 574, follow-up 24 and 52 weeks), OSKIRA-1 (N = 131, follow-up 24 weeks)). Two osteoarthritis (OA) studies (SMC01 (N = 447), SMC02 (N = 81)) were included as non-RA comparators. Specific tissue-derived neoepitopes measured at baseline, included: C2M (cartilage degradation); CTX-I and PINP (bone turnover); C1M and C3M (interstitial matrix degradation); CRPM (CRP metabolite) and VICM (macrophage activity). Clustering was performed to identify putative endotypes. We identified five clusters (A-E). Clusters A and B were characterized by generally higher levels of biomarkers than other clusters, except VICM which was significantly higher in cluster B than in cluster A (p<0.001). Biomarker levels in Cluster C were all close to the median, whilst Cluster D was characterised by low levels of all biomarkers. Cluster E also had low levels of most biomarkers, but with significantly higher levels of CTX-I compared to cluster D. There was a significant difference in ΔSHP score observed at 52 weeks (p<0.05). We describe putative RA endotypes based on biomarkers reflecting joint tissue metabolism. These endotypes differ in their underlining pathogenesis, and may in the future have utility for patient treatment selection.


A Prospective, Randomized, Open-Label Trial of Early versus Late Favipiravir Therapy in Hospitalized Patients with COVID-19.
Favipiravir is an oral broad-spectrum inhibitor of viral RNA-dependent RNA polymerase that is approved for treatment of influenza in Japan. We conducted a prospective, randomized, open-label, multicenter trial of favipiravir for the treatment of COVID-19 at 25 hospitals across Japan. Eligible patients were adolescents and adults admitted with COVID-19 who were asymptomatic or mildly ill and had an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. Patients were randomly assigned at a 1:1 ratio to early or late favipiravir therapy (in the latter case, the same regimen starting on day 6 instead of day 1). The primary endpoint was viral clearance by day 6. The secondary endpoint was change in viral load by day 6. Exploratory endpoints included time to defervescence and resolution of symptoms. Eighty-nine patients were enrolled, of whom 69 were virologically evaluable. Viral clearance occurred within 6 days in 66.7% and 56.1% of the early and late treatment groups (adjusted hazard ratio [aHR], 1.42; 95% confidence interval [95% CI], 0.76 to 2.62). Of 30 patients who had a fever (≥37.5°C) on day 1, times to defervescence were 2.1 days and 3.2 days in the early and late treatment groups (aHR, 1.88; 95% CI, 0.81 to 4.35). During therapy, 84.1% developed transient hyperuricemia. Favipiravir did not significantly improve viral clearance as measured by reverse transcription-PCR (RT-PCR) by day 6 but was associated with numerical reduction in time to defervescence. Neither disease progression nor death occurred in any of the patients in either treatment group during the 28-day participation. (This study has been registered with the Japan Registry of Clinical Trials under number jRCTs041190120.).


Ancient origin and gene mosaicism of the progenitor of Mycobacterium tuberculosis.
The highly successful human pathogen Mycobacterium tuberculosis has an extremely low level of genetic variation, which suggests that the entire population resulted from clonal expansion following an evolutionary bottleneck around 35,000 y ago. Here, we show that this population constitutes just the visible tip of a much broader progenitor species, whose extant representatives are human isolates of tubercle bacilli from East Africa. In these isolates, we detected incongruence among gene phylogenies as well as mosaic gene sequences, whose individual elements are retrieved in classical M. tuberculosis. Therefore, despite its apparent homogeneity, the M. tuberculosis genome appears to be a composite assembly resulting from horizontal gene transfer events predating clonal expansion. The amount of synonymous nucleotide variation in housekeeping genes suggests that tubercle bacilli were contemporaneous with early hominids in East Africa, and have thus been coevolving with their human host much longer than previously thought. These results open novel perspectives for unraveling the molecular bases of M. tuberculosis evolutionary success.


The tubby family proteins.
The tubby mouse shows a tripartite syndrome characterized by maturity-onset obesity, blindness and deafness. The causative gene Tub is the founding member of a family of related proteins present throughout the animal and plant kingdoms, each characterized by a signature carboxy-terminal tubby domain. This domain consists of a β barrel enclosing a central α helix and binds selectively to specific membrane phosphoinositides. The vertebrate family of tubby-like proteins (TULPs) includes the founding member TUB and the related TULPs, TULP1 to TULP4. Tulp1 is expressed in the retina and mutations in TULP1 cause retinitis pigmentosa in humans; Tulp3 is expressed ubiquitously in the mouse embryo and is important in sonic hedgehog (Shh)-mediated dorso-ventral patterning of the spinal cord. The amino terminus of these proteins is diverse and directs distinct functions. In the best-characterized example, the TULP3 amino terminus binds to the IFT-A complex, a complex important in intraflagellar transport in the primary cilia, through a short conserved domain. Thus, the tubby family proteins seem to serve as bipartite bridges through their phosphoinositide-binding tubby and unique amino-terminal functional domains, coordinating multiple signaling pathways, including ciliary G-protein-coupled receptor trafficking and Shh signaling. Molecular studies on this functionally diverse protein family are beginning to provide us with remarkable insights into the tubby-mouse syndrome and other related diseases.


SU-E-T-04: Repair Kinetics of Sublethal Damage in Rat Cervical Spinal Cord - Application of the GLQ Model Incorporating Reciprocal Time Pattern.
The repair rate conventionally assumed to be constant during the entire radiation course. However, increasing evidences from animal studies show that the repair process may slow down with time and the experimental data does not fit an exponential pattern. To address this enigma, we presented a generalized linear-quadratic (gLQ) model incorporating reciprocal time repair pattern of sublethal damage and fit the model to published experimental data. In the gLQ model, the parameter G in the surviving fraction represents the repair process of sublethal damage with Tr as the repair half-time. When a reciprocal time pattern of repair process was adopted, a generalized form of G was derived analytically for arbitrary radiation schemes (I(t) = Ii, ti-1 < t < ti, where i=1 to N, Ii is the dose rate of the ith segment to the voxel in question, and N is the total number of segments for the entire treatment course). The published animal data was used to test the reciprocal formulas. The gLQ model incorporating a reciprocal time pattern to describe the repair process was used to fit the experimental data. The reciprocal model showed better fit to the animal data than the exponential model, particularly for the ED50 data, resulting in the following parameters: the reduced chi-square of 1.52, α/β=0.9 Gy, Tr =1.0 h for the reciprocal model and reduced chi-square of 3.39, α/β=1.6 Gy, Tr=3.5 h for the exponential model. The modeling results suggest that the gLQ model incorporating the reciprocal time pattern of sublethal damage repair facilitates the interpretation of complex experimental designs, especially for irregular dose delivery schedules. These formulas can be used to analyze experimental and clinical data, where a slowing-down of the repair process occurs during the radiation therapy course.


Relationship of serum IgE concentration to level and rate of decline of pulmonary function: the Normative Aging Study.
Previous reports on the relationship between serum immunoglobulin E (IgE) concentration and the level and rate of decline of pulmonary function in the general population have produced conflicting results. The relationship between total serum IgE concentration and pulmonary function was therefore examined in 1078 men aged 41-86 years followed in the Normative Aging Study. The serum IgE concentration determined at the start of the three year follow up period was examined in relation to both the level and longitudinal rate of decline of forced expiratory volume in one second (FEV1), forced vital capacity (FVC), and FEV1/FVC. In a cross sectional analysis restricted to subjects who had ever smoked cigarettes, multiple linear regression models indicated an inverse association between total serum IgE concentration and both FEV1 (beta = -0.090 1/log10 IU/ml; SE = 0.030; p < 0.005) and FVC (beta = -0.110 1/log10 IU/ml; SE = 0.034; p < 0.005) but not FEV1/FVC, after adjustment for age and height. This relationship persisted when individuals with diagnosed asthma or methacholine hyperresponsiveness were excluded. In subjects who had never smoked cigarettes the total serum IgE concentration was unrelated to spirometric indices. No association was observed in smokers or non-smokers between the serum IgE concentration measured at the beginning of the period of follow up and the decline in FEV1, FVC, or FEV1/FVC. Increased levels of serum IgE measured at the beginning of the follow up period are associated with lower levels of pulmonary function but are not predictive of an accelerated rate in the decline of pulmonary function among middle aged and older men.


Cannabinoid and kappa opioid receptors reduce potassium K current via activation of G(s) proteins in cultured hippocampal neurons.
The current study showed that potassium K current (I(K)), which is evoked at depolarizing potentials between -30 and +40 mV in cultured hippocampal neurons, was significantly reduced by exposure to the CB1 cannabinoid receptor agonist WIN 55,212-2 (WIN-2). WIN-2 (20-40 nM) produced an average 45% decrease in I(K) amplitude across all voltage steps, which was prevented by SR141716A, the CB1 receptor antagonist. The cannabinoid receptor has previously been shown to be G(i/o) protein-linked to several cellular processes; however, the decrease in I(K) was unaffected by modulators of G(i/o) proteins and agents that alter levels of protein kinase A. In contrast, CB1 receptor-mediated or direct activation of G(s) proteins with cholera toxin (CTX) produced the same decrease in I(K) amplitude as WIN-2, and the latter was blocked in CTX-treated cells. G(s) protein inhibition via GDPbetaS also eliminated the effects of WIN-2 on I(K). Consistent with this outcome, activation of protein kinase C (PKC) by arachidonic acid produced similar effects to WIN-2 and CTX. Kappa opioid receptor agonists, which also reduce I(K) amplitude via G(s) proteins, were compared with WIN-2 actions on I(K.) The kappa receptor agonist U50,488 reduced I(K) amplitude in the same manner as WIN-2, while the kappa receptor antagonist, nor-binaltorphimine, actually increased I(K) amplitude and significantly reduced the effect of co-administered WIN-2. The results indicate that CB1 and kappa receptor activation is additive with respect to I(K) amplitude, suggesting that CB1 and kappa receptors share a common G(s) protein signaling pathway involving PKC.


Sucrose accumulation in sweet sorghum stems occurs by apoplasmic phloem unloading and does not involve differential Sucrose transporter expression.
Sorghum (Sorghum bicolor L. Moench) cultivars store non-structural carbohydrates predominantly as either starch in seeds (grain sorghums) or sugars in stems (sweet sorghums). Previous research determined that sucrose accumulation in sweet sorghum stems was not correlated with the activities of enzymes functioning in sucrose metabolism, and that an apoplasmic transport step may be involved in stem sucrose accumulation. However, the sucrose unloading pathway from stem phloem to storage parenchyma cells remains unelucidated. Sucrose transporters (SUTs) transport sucrose across membranes, and have been proposed to function in sucrose partitioning differences between sweet and grain sorghums. The purpose of this study was to characterize the key differences in carbohydrate accumulation between a sweet and a grain sorghum, to define the path sucrose may follow for accumulation in sorghum stems, and to determine the roles played by sorghum SUTs in stem sucrose accumulation. Dye tracer studies to determine the sucrose transport route revealed that, for both the sweet sorghum cultivar Wray and grain sorghum cultivar Macia, the phloem in the stem veins was symplasmically isolated from surrounding cells, suggesting sucrose was apoplasmically unloaded. Once in the phloem apoplasm, a soluble tracer diffused from the vein to stem parenchyma cell walls, indicating the lignified mestome sheath encompassing the vein did not prevent apoplasmic flux outside of the vein. To characterize carbohydrate partitioning differences between Wray and Macia, we compared the growth, stem juice volume, solute contents, SbSUTs gene expression, and additional traits. Contrary to previous findings, we detected no significant differences in SbSUTs gene expression within stem tissues. Phloem sieve tubes within sweet and grain sorghum stems are symplasmically isolated from surrounding cells; hence, unloading from the phloem likely occurs apoplasmically, thereby defining the location of the previously postulated step for sucrose transport. Additionally, no changes in SbSUTs gene expression were detected in sweet vs. grain sorghum stems, suggesting alterations in SbSUT transcript levels do not account for the carbohydrate partitioning differences between cultivars. A model illustrating sucrose phloem unloading and movement to stem storage parenchyma, and highlighting roles for sucrose transport proteins in sorghum stems is discussed.


Transient receptor potential ankyrin 1 is expressed by inhibitory motoneurons of the mouse intestine.
Transient receptor potential ankyrin (TRPA) 1, an excitatory ion channel expressed by sensory neurons, mediates somatic and visceral pain in response to direct activation or noxious mechanical stimulation. Although the intestine is routinely exposed to irritant alimentary compounds and inflammatory mediators that activate TRPA1, there is no direct evidence for functional TRPA1 receptors on enteric neurons, and the effects of TRPA1 activation on intestinal function have not been determined. We characterized expression of TRPA1 by enteric neurons and determined its involvement in the control of intestinal contractility and transit. TRPA1 expression was characterized by reverse-transcription polymerase chain reaction and immunofluorescence analyses. TRPA1 function was examined by Ca(2+) imaging and by assays of contractile activity and transit. We detected TRPA1 messenger RNA in the mouse intestine and TRPA1 immunoreactivity in enteric neurons. The cecum and colon had immunoreactivity for neuronal TRPA1, but the duodenum did not. TRPA1 immunoreactivity was also detected in inhibitory motoneurons and descending interneurons, cholinergic neurons, and intrinsic primary afferent neurons. TRPA1 activators, including cinnamaldehyde, allyl isothiocyanate (AITC), and 4-hydroxynonenal, increased [Ca(2+)](i) in myenteric neurons. These were reduced by a TRPA1 antagonist (HC-030031) or deletion of Trpa1. TRPA1 activation inhibited contractility of the segments of colon but not stomach or small intestine of Trpa1(+/+) but not Trpa1(-/-) mice; this effect was reduced by tetrodotoxin or N(G)-nitro-l-arginine methyl ester. Administration of AITC by gavage did not alter gastric emptying or small intestinal transit, but luminal AITC inhibited colonic transit via TRPA1. Functional TRPA1 is expressed by enteric neurons, and activation of neuronal TRPA1 inhibits spontaneous neurogenic contractions and transit of the colon.


Information transduction capacity of noisy biochemical signaling networks.
Molecular noise restricts the ability of an individual cell to resolve input signals of different strengths and gather information about the external environment. Transmitting information through complex signaling networks with redundancies can overcome this limitation. We developed an integrative theoretical and experimental framework, based on the formalism of information theory, to quantitatively predict and measure the amount of information transduced by molecular and cellular networks. Analyzing tumor necrosis factor (TNF) signaling revealed that individual TNF signaling pathways transduce information sufficient for accurate binary decisions, and an upstream bottleneck limits the information gained via multiple integrated pathways. Negative feedback to this bottleneck could both alleviate and enhance its limiting effect, despite decreasing noise. Bottlenecks likewise constrain information attained by networks signaling through multiple genes or cells.