Seasonal dynamics of mobile carbon supply in Quercus aquifolioides at the upper elevational limit.
Many studies have tried to explain the physiological mechanisms of the alpine treeline phenomenon, but the debate on the alpine treeline formation remains controversial due to opposite results from different studies. The present study explored the carbon-physiology of an alpine shrub species (Quercus aquifolioides) grown at its upper elevational limit compared to lower elevations, to test whether the elevational limit of alpine shrubs (<3 m in height) are determined by carbon limitation or growth limitation. We studied the seasonal variations in non-structural carbohydrate (NSC) and its pool size in Q. aquifolioides grown at 3000 m, 3500 m, and at its elevational limit of 3950 m above sea level (a.s.l.) on Zheduo Mt., SW China. The tissue NSC concentrations along the elevational gradient varied significantly with season, reflecting the season-dependent carbon balance. The NSC levels in tissues were lowest at the beginning of the growing season, indicating that plants used the winter reserve storage for re-growth in the early spring. During the growing season, plants grown at the elevational limit did not show lower NSC concentrations compared to plants at lower elevations, but during the winter season, storage tissues, especially roots, had significantly lower NSC concentrations in plants at the elevational limit compared to lower elevations. The present results suggest the significance of winter reserve in storage tissues, which may determine the winter survival and early-spring re-growth of Q. aquifolioides shrubs at high elevation, leading to the formation of the uppermost distribution limit. This result is consistent with a recent hypothesis for the alpine treeline formation.


Predictive value of the Diverticular Inflammation and Complication Assessment (DICA) endoscopic classification on the outcome of diverticular disease of the colon: An international study.
Diverticular Inflammation and Complication Assessment (DICA) endoscopic classification has been recently developed for patients suffering from diverticulosis and diverticular disease. We assessed retrospectively the predictive value of DICA in patients for whom endoscopic data and clinical follow-up were available. For each patient, we recorded: age, severity of DICA, presence of abdominal pain, C-reactive protein and faecal calprotectin test (if available) at the time of diagnosis; months of follow-up; therapy taken during the follow-up to maintain remission (if any); occurrence/recurrence of diverticulitis; need of surgery. We enrolled 1651 patients (793 M, 858 F, mean age 66.6 ± 11.1 years): 939 (56.9%) patients were classified as DICA 1, 501 (30.3%) patients as DICA 2 and 211 (12.8%) patients as DICA 3. The median follow-up was 24 (9-38) months. Acute diverticulitis (AD) occurred/recurred in 263 (15.9%) patients; surgery was necessary in 57 (21.7%) cases. DICA was the only factor significantly associated to the occurrence/recurrence of diverticulitis and surgery either at univariate (χ(2 )= 405.029; p < 0.0001) or multivariate analysis (hazard ratio = 4.319, 95% confidence interval (CI) 3.639-5.126; p < 0.0001). Only in DICA 2 patients was therapy effective for prevention of AD occurrence/recurrence with a hazard ratio (95% CI) of 0.598 (0.391-0.914) (p = 0.006, log rank test). Mesalazine-based therapies reduced the risk of AD occurrence/recurrence and needs of surgery with a hazard ratio (95% CI) of 0.2103 (0.122-0.364) and 0.459 (0.258-0.818), respectively. DICA classification is a valid parameter to predict the risk of diverticulitis occurrence/recurrence in patients suffering from diverticular disease of the colon.


Capturing the most wanted taxa through cross-sample correlations.
The Human Microbiome Project (HMP) identified the 16S rRNA gene sequences of 'most wanted' taxa-prevalent in the healthy human microbiota but distant from previously known sequences. Since 2012, few of the corresponding genomes have been isolated and sequenced, and only through advanced isolation techniques. We demonstrate that the genomes of the most wanted taxa can be identified computationally through their correlation in abundance across multiple public metagenomic data sets. We link over 200 most wanted sequences with nearly complete genome sequences, including half of the taxa identified as high-priority targets by the HMP. The genomes we identify have strong similarity to genomes reconstructed through expensive isolation techniques, and provide a more complete functional characterization of these organisms than can be extrapolated from their 16S rRNA gene. We also provide insights into the function of organisms for which 16S rRNA gene signatures were recently reported to be associated with health and host genetic factors.


Automated registration of magnetic resonance imaging and optoacoustic tomography data for experimental studies.
Multimodal imaging combining optoacoustic tomography (OAT) with magnetic resonance imaging (MRI) enables spatiotemporal resolution complementarity, improves accurate quantification, and thus yields more insights into physiology and pathophysiology. However, only manual landmark based coregistration of OAT-MRI has been used so far. We developed a toolbox (RegOA), which frames an automated registration pipeline to align OAT with high-field MR images based on mutual information. We assessed the performance of the registration method using images acquired on one phantom with fiducial markers and in vivo/ex vivo data of mouse heads/brain. The accuracy and robustness of the registration are improved using a two-step registration method with preprocessing of OAT and MRI data. The major advantages of our approach are minimal user input and quantitative assessment of the registration error. The registration with MR and standard reference atlas enables regional information extraction, facilitating the accurate, objective, and rapid analysis of large groups of rodent OAT and MR images.


A search algorithm for identifying likely users and non-users of marijuana from the free text of the electronic medical record.
The harmful effects of marijuana on health and in particular cardiovascular health are understudied. To develop such knowledge, an efficient method of developing an informative cohort of marijuana users and non-users is needed. We identified patients with a diagnosis of coronary artery disease using ICD-9 codes who were seen in the San Francisco VA in 2015. We imported these patients' medical record notes into an informatics platform that facilitated text searches. We categorized patients into those with evidence of marijuana use in the past 12 months and patients with no such evidence, using the following text strings: "marijuana", "mjx", and "cannabis". We randomly selected 51 users and 51 non-users based on this preliminary classification, and sent a recruitment letter to 97 of these patients who had contact information available. Patients were interviewed on marijuana use and domains related to cardiovascular health. Data on marijuana use collected from the medical record was compared to data collected as part of the interview. The interview completion rate was 71%. Among the 35 patients identified by text strings as having used marijuana in the previous year, 15 had used marijuana in the past 30 days (positive predictive value = 42.9%). The probability of use in the past month increased from 8.8% to 42.9% in people who have these keywords in their medical record compared to those who did not have these terms in their medical record. Methods that combine text search strategies for participant recruitment with health interviews provide an efficient approach to developing prospective cohorts that can be used to study the health effects of marijuana.


Effects of a simulated martian UV flux on the cyanobacterium, Chroococcidiopsis sp. 029.
Dried monolayers of Chroococcidiopsis sp. 029, a desiccation-tolerant, endolithic cyanobacterium, were exposed to a simulated martian-surface UV and visible light flux, which may also approximate to the worst-case scenario for the Archean Earth. After 5 min, there was a 99% loss of cell viability, and there were no survivors after 30 min. However, this survival was approximately 10 times higher than that previously reported for Bacillus subtilis. We show that under 1 mm of rock, Chroococcidiopsis sp. could survive (and potentially grow) under the high martian UV flux if water and nutrient requirements for growth were met. In isolated cells, phycobilisomes and esterases remained intact hours after viability was lost. Esterase activity was reduced by 99% after a 1-h exposure, while 99% loss of autofluorescence required a 4-h exposure. However, cell morphology was not changed, and DNA was still detectable by 4',6-diamidino-2-phenylindole staining after an 8-h exposure (equivalent to approximately 1 day on Mars at the equator). Under 1 mm of simulant martian soil or gneiss, the effect of UV radiation could not be detected on esterase activity or autofluorescence after 4 h. These results show that under the intense martian UV flux the morphological signatures of life can persist even after viability, enzymatic activity, and pigmentation have been destroyed. Finally, the global dispersal of viable, isolated cells of even this desiccation-tolerant, ionizing-radiation-resistant microorganism on Mars is unlikely as they are killed quickly by unattenuated UV radiation when in a desiccated state. These findings have implications for the survival of diverse microbial contaminants dispersed during the course of human exploratory class missions on the surface of Mars.


Validation of the surveillance system for new cases of tuberculosis in a province of northern Italy. Varese Tuberculosis Study Group.
In Italy tuberculosis (TB) surveillance is hampered by several problems (inconsistency of data and underreporting). A tuberculosis surveillance system was established in Varese Province (VSS) in 1992. The aim of the present study was, using VSS, to estimate: 1) coverage and validity of data collected by the national Compulsory Surveillance System (CSS); 2) validity of diagnosis and risk factors for tuberculosis; 3) tuberculosis incidence. A specifically designed form, including the same items as the CSS form and 10 more questions on risk factors and diagnostic issue, was completed in all the existing health facilities in the province for each new tuberculosis case diagnosed, collected by the study co-ordinator during monthly supervision after a review of clinical records, and stored in a database at the co-ordinating centre. Routine notifications collected at national level by CSS for the province were nominally linked with VSS data. To evaluate coverage and data validity, a comparison was made between VSS and CSS data after removal of duplicates. Quality of data, risk factors and incidence were evaluated on VSS data (gold standard). CSS coverage was 63% with a significant underreporting of extrapulmonary cases. Within CSS, 20% of data were missing (particularly diagnostic information: 42-72%). According to VSS, smear was performed on 88% of cases and culture on 66%. Half of the cases had no risk factors. The overall incidence of tuberculosis in Varese Province was 15 per 100,000. We estimated the coverage and validity of the data collected by CSS, validity of diagnosis, risk factors for and incidence of tuberculosis and proposed several suggestions to improve CSS nationwide.


Assessing pearl quality using reflectance UV-Vis spectroscopy: does the same donor produce consistent pearl quality?
Two groups of commercial quality ("acceptable") pearls produced using two donors, and a group of "acceptable" pearls from other donors were analyzed using reflectance UV-Vis spectrophotometry. Three pearls with different colors produced by the same donor showed different absorption spectra. Cream and gold colored pearls showed a wide absorption from 320 to about 460 nm, while there was just slight reflectance around 400 nm by the white pearl with a pink overtone. Cream and gold pearls reached a reflectance peak at 560 to 590 nm, while the white pearl with pink overtone showed slightly wider absorption in this region. Both cream and gold pearls showed an absorption peak after the reflectance peak, at about 700 nm for the cream pearl and 750 nm for the gold pearl. Two other pearls produced by the same donor (white with cream overtone and cream with various overtones) showed similar spectra, which differed in their intensity. One of these pearls had very high lustre and its spectrum showed a much higher percentage reflectance than the second pearl with inferior lustre. This result may indicate that reflectance is a useful quantitative indicator of pearl lustre. The spectra of two white pearls resulting from different donors with the same color nacre (silver) showed a reflectance at 260 nm, followed by absorption at 280 nm and another reflectance peak at 340 nm. After this peak the spectra for these pearls remained flat until a slight absorption peak around 700 nm. Throughout the visible region, all white pearls used in this study showed similar reflectance spectra although there were differences in reflectance intensity. Unlike the spectral results from white pearls, the results from yellow and gold pearls varied according to color saturation of the pearl. The results of this study show that similarities between absorption and reflectance spectra of cultured pearls resulting from the same saibo donor are negligible and could not be detected with UV-Vis spectrophotometry. Nevertheless, this technique could have a role to play in developing less subjective methods of assessing pearl quality and in further studies of the relationships between pearl quality and that of the donor and recipient oysters.


Cancer mortality and serum levels of carotenoids, retinol, and tocopherol: a population-based follow-up study of inhabitants of a rural area of Japan.
A total of 3,182 subjects (1,239 males and 1,943 females) aged from 39y to 79y, were recruited from the inhabitants of a rural area in Japan who participated in health check-up programs from 1988 to 1995. During the 10.5 year follow-up, 287 deaths (175 males and 112 females) from all causes, 134 (81 males and 53 females) from cancer of all sites, 31 from lung cancer, 21 from colorectal cancer, 20 from stomach cancer, and 62 from other cancers, were identified among the cohort subjects. Fasting serum samples were taken at the time of the health check-ups, and serum levels of carotenoids, retinol and tocopherols were separately determined by HPLC. Statistical analyses were performed using Cox's proportional hazard model after adjusting for sex, age, and other confounding factors. High serum levels of alpha- and beta- carotenes and lycopene were found to marginally significantly or significantly reduce the risk for mortality rates of cancer of all sites and of colorectal cancers. High serum levels of beta-cryptoxanthin also showed an inversely relation with the risk of mortality from lung and stomach cancers, but this was not statistically significant. High intake of green-yellow vegetables contributing to serum levels of alpha- and beta- carotenes, as well as lycopene, may reduce the risk of cancer mortality, especially from colorectal cancer, in rural Japanese.


Interhemispheric transmission of visuomotor information in humans: fMRI evidence.
Normal human subjects underwent functional magnetic resonance imaging (fMRI) while performing a simple visual manual reaction-time (RT) task with lateralized brief stimuli, the so-called Poffenberger's paradigm. This paradigm was employed to measure interhemispheric transmission (IT) time by subtracting mean RT for the uncrossed hemifield-hand conditions, that is, those conditions not requiring an IT, from the crossed hemifield-hand conditions, that is, those conditions requiring an IT to relay visual information from the hemisphere of entry to the hemisphere subserving the response. The obtained difference is widely believed to reflect callosal conduction time, but so far there is no direct physiological evidence in humans. The aim of our experiment was twofold: first, to test the hypothesis that IT of visuomotor information requires the corpus callosum and to identify the cortical areas specifically activated during IT. Second, we sought to discover whether IT occurs mainly at premotor or perceptual stages of information processing. We found significant activations in a number of frontal, parietal, and temporal cortical areas and in the genu of the corpus callosum. These activations were present only in the crossed conditions and therefore were specifically related to IT. No selective activation was present in the uncrossed conditions. The location of the activated callosal and cortical areas suggests that IT occurs mainly, but not exclusively, at premotor level. These results provide clear cut evidence in favor of the hypothesis that the crossed-uncrossed difference in the Poffenberger paradigm depends on IT rather than on a differential hemispheric activation.


Cis-regulatory control of corticospinal system development and evolution.
The co-emergence of a six-layered cerebral neocortex and its corticospinal output system is one of the evolutionary hallmarks of mammals. However, the genetic programs that underlie their development and evolution remain poorly understood. Here we identify a conserved non-exonic element (E4) that acts as a cortex-specific enhancer for the nearby gene Fezf2 (also known as Fezl and Zfp312), which is required for the specification of corticospinal neuron identity and connectivity. We find that SOX4 and SOX11 functionally compete with the repressor SOX5 in the transactivation of E4. Cortex-specific double deletion of Sox4 and Sox11 leads to the loss of Fezf2 expression, failed specification of corticospinal neurons and, independent of Fezf2, a reeler-like inversion of layers. We show evidence supporting the emergence of functional SOX-binding sites in E4 during tetrapod evolution, and their subsequent stabilization in mammals and possibly amniotes. These findings reveal that SOX transcription factors converge onto a cis-acting element of Fezf2 and form critical components of a regulatory network controlling the identity and connectivity of corticospinal neurons.


Adolescent diet and incidence of proliferative benign breast disease.
Studies of adult diet and risk of breast cancer have yielded mainly null results, but this does not rule out a possible impact of adolescent diet. This study examined associations between components of adolescent diet and risk of proliferative benign breast disease (BBD), a marker for breast cancer. The study population consisted of 29494 women in the Nurses' Health Study II who completed a questionnaire on adolescent diet in 1998 and who were 33-53 years of age at that time. A total of 470 new cases of proliferative BBD were identified between 1991 and 1997. Incidence rate ratios (RRs) and 95% confidence intervals (CIs) were calculated for quartiles of energy-adjusted intakes, using the lowest quartile of each as the reference group. Total fat intake during adolescence was unrelated to risk of proliferative BBD, although there were positive associations for intakes of animal fat and monounsaturated fat and an inverse association for intake of vegetable fat. For vitamin E intake, the multivariate RRs were 1.13, 0.88, and 0.79 (95% CI, 0.61-1.04) for women in the second, third, and highest quartiles, respectively (P for trend = 0.05). The multivariate RRs were 0.94, 0.99, and 0.75 (95% CI, 0.57-0.98) for women in increasing quartiles of fiber intake (P for trend = 0.05). Vegetable fat, vitamin E, and fiber intakes during adolescence were inversely associated with risk of proliferative BBD in this population. Confirmation of these associations may suggest a means for prevention of breast cancer.


Patients' understanding of cellulitis and views about how best to prevent recurrent episodes: mixed-methods study in primary and secondary care.
Cellulitis is a common painful infection of the skin and underlying tissues that recurs in approximately one-third of cases. The only proven strategy to reduce the risk of recurrence is long-term, low-dose antibiotics. Given current concerns about antibiotic resistance and the pressure to reduce antibiotic prescribing, other prevention strategies are needed. To explore patients' views about cellulitis and different ways of preventing recurrent episodes. Adults aged ≥ 18 years with a history of first-episode or recurrent cellulitis were invited through primary care, hospitals and advertising to complete a survey, take part in an interview or both. Thirty interviews were conducted between August 2016 and July 2017. Two hundred and forty surveys were completed (response rate 17%). Triangulation of quantitative and qualitative data showed that people who have had cellulitis have wide-ranging beliefs about what can cause cellulitis and are often unaware of risk of recurrence or potential strategies to prevent recurrence. Enhanced foot hygiene, applying emollients daily, exercise and losing weight were more popular potential strategies than the use of compression stockings or long-term antibiotics. Participants expressed caution about long-term oral antibiotics, particularly those who had experienced only one episode of cellulitis. People who have had cellulitis are keen to know about possible ways to prevent further episodes. Enhanced foot hygiene, applying emollients daily, exercise and losing weight were generally viewed to be more acceptable, feasible strategies than compression or antibiotics, but further research is needed to explore uptake and effectiveness in practice.


Anaplasma marginale and A. phagocytophilum in cattle in Tunisia.
Tick-borne diseases caused by Anaplasma species put serious constraints on the health and production of domestic cattle in tropical and sub-tropical regions. After recovering from a primary infection, cattle typically become persistent carriers of pathogens and play a critical role in the epidemiology of the disease, acting as reservoirs of the Anaplasma spp. In this study a duplex PCR assay was used for the simultaneous detection of Anaplasma marginale and Anaplasma phagocytophilum in cattle using two primer pairs targeting msp4 and msp2 genes, respectively. We used this method to analyze DNA preparations derived from 328 blood cattle samples that were collected from 80 farms distributed among Tunisia's four bioclimatic zones. The prevalence of the A. marginale infection (24.7 %) was significantly higher and more widespread (in all bioclimatic areas) than that of A. phagocytophilum (0.6 %), which was found in a mixed infection with A. marginale. The duplex PCR assay used proved to be a rapid, specific and inexpensive mean for the simultaneous detection of Anaplasma marginale and Anaplasma phagocytophilum in cattle blood. It allowed us to report the identification of A. phagocytophilum for the first time in cattle in Tunisia and confirm the presence of A. marginale in cattle from several geographical areas of the country. Further epidemiological studies undertaken using this assay will help improve the surveillance of the associated diseases in the regions where they are endemic.


Mechanistic Investigation on the Formation of 2-Halo-3-aryl-4(3H)-quinazoliniminium Halides from Heteroenyne-allenes: A Computational Study.
Intramolecular cyclization of the heteroenyne-allene, 2-((phenylimino)methyleneamino)-benzonitrile (1) in the presence of HCl to produce 2-chloro-3-phenyl-4(3H)-quinazoliniminium chloride (Qz) involves the formation of two new bonds: a C-Cl bond and a C-N bond. We propose five pathways for this reaction. Four of these pathways involve chloride capture to form the C-Cl bond prior to the intramolecular nucleophilic attack to form the C-N bond, while one pathway involves ring closure to form the C-N bond prior to C-Cl bond formation. All calculations were carried out at B3LYP and MP2 levels of theory and employed the 6-311G* basis set. The solvent effects were considered using a Polarized Continuum Model with dichloromethane as the solvent. The calculations at both levels show that the mechanism involves initial protonation of 1, preferentially at Nα to give 2 which rapidly captures the chloride ion to form 5. This intermediate is protonated at the -CN group to form 8ROT, which then tautomerizes to its more stable isomer 9ROT. The latter undergoes intramolecular nucleophilic attack from Nβ to the protonated -CN group to form the cyclized intermediate 12, which tautomerizes to its most stable isomer 13. The coordination of Cl(-) ion present in the solution with 13 gives the final product Qz.


Survival of Enterococcus faecalis in infected dentinal tubules after root canal filling with different root canal sealers in vitro.
To investigate the ability of different endodontic sealers and calcium hydroxide to kill bacteria in experimentally infected dentinal tubules. Fifty-six human root segments were enlarged to size 2 (ISO size 090) Largo Peeso Reamer. After treatment with 17% EDTA and 5% NaOCl for 4 min each, the specimens were infected with Enterococcus faecalis for 3 weeks. The roots were divided into eight groups and filled with gutta-percha and AH Plus (AH); Grossman's sealer (GS); Ketac-Endo (KE); Apexit (AP); RoekoSeal Automix (RSA); or RoekoSeal Automix with an experimental primer (RP), or calcium hydroxide (CH) only. One group of specimens was left unfilled for control (CT). Following storage in humid conditions at 37 degrees C for 7 days, the root canals were re-established with new sterile Largo size 2. Dentine samples from each canal were then collected using a sterile size 5 (ISO size 150) Largo Peeso Reamer. The number of colony-forming units (CFU) was determined for each sample. The mean log10 CFU in all test groups was significantly lower (P < 0.05) than that in the CT group. Root filling with AH and GS killed bacteria (mean CFU = 0) in the dentinal tubules. The mean log10 CFU for the CH group (0.53) was lower than that of RSA, AP, RP and KE (1.36, 1.40, 1.46 and 1.94, respectively), but only the difference between the CH and the KE groups was statistically significant (P < 0.05). Root fillings in vitro with gutta-percha and AH or GS were effective in killing E. faecalis in dentinal tubules. Other endodontic sealers, as well as CH, were less effective.


Ginkgo biloba prevents mobile phone-induced oxidative stress in rat brain.
The widespread use of mobile phones (MP) in recent years has raised the research activities in many countries to determine the consequences of exposure to the low-intensity electromagnetic radiation (EMR) of mobile phones. Since several experimental studies suggest a role of reactive oxygen species (ROS) in EMR-induced oxidative damage in tissues, in this study, we investigated the effect of Ginkgo biloba (Gb) on MP-induced oxidative damage in brain tissue of rats. Rats (EMR+) were exposed to 900 MHz EMR from MP for 7 days (1 h/day). In the EMR+Gb groups, rats were exposed to EMR and pretreated with Gb. Control and Gb-administrated groups were produced by turning off the mobile phone while the animals were in the same exposure conditions. Subsequently, oxidative stress markers and pathological changes in brain tissue were examined for each groups. Oxidative damage was evident by the: (i) increase in malondialdehyde (MDA) and nitric oxide (NO) levels in brain tissue, (ii) decrease in brain superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and (iii) increase in brain xanthine oxidase (XO) and adenosine deaminase (ADA) activities. These alterations were prevented by Gb treatment. Furthermore, Gb prevented the MP-induced cellular injury in brain tissue histopathologically. Reactive oxygen species may play a role in the mechanism that has been proposed to explain the biological side effects of MP, and Gb prevents the MP-induced oxidative stress to preserve antioxidant enzymes activity in brain tissue.


Stress sensitivity in metastatic breast cancer: analysis of hypothalamic-pituitary-adrenal axis function.
The normal diurnal cortisol cycle has a peak in the morning, decreasing rapidly over the day, with low levels during the night, then rising rapidly again to the morning peak. A pattern of flatter daytime slopes has been associated with more rapid cancer progression in both animals and humans. We studied the relationship between the daytime slopes and other daytime cortisol responses to both pharmacological and psychosocial challenges of hypothalamic-pituitary-adrenal (HPA) axis function as well as DHEA in a sample of 99 women with metastatic breast cancer, in hopes of elucidating the dysregulatory process. We found that the different components of HPA regulation: the daytime cortisol slope, the rise in cortisol from waking to 30 min later, and cortisol response to various challenges, including dexamethasone (DEX) suppression, corticotrophin releasing factor (CRF) activation, and the Trier Social Stress Task, were at best modestly associated. Escape from suppression stimulated by 1mg of DEX administered the night before was moderately but significantly associated with flatter daytime cortisol slopes (r=0.28 to .30 at different times of the post DEX administration day, all p<.01). Daytime cortisol slopes were also moderately but significant associated with the rise in cortisol from waking to 30 min after awakening (r=.29, p=.004, N=96), but not with waking cortisol level (r=-0.13, p=.19). However, we could not detect any association between daytime cortisol slope and activation of cortisol secretion by either CRF infusion or the Trier Social Stress Task. The CRF activation test (following 1.5mg of DEX to assure that the effect was due to exogenous CRF) produced ACTH levels that were correlated (r=0.66, p<.0001, N=74) with serum cortisol levels, indicating adrenal responsiveness to ACTH stimulation. Daytime cortisol slopes were significantly correlated with the slope of DHEA (r=.21, p=.04, N=95). Our general findings suggest that flatter daytime cortisol slopes among metastatic breast cancer patients may be related to disrupted feedback inhibition rather than hypersensitivity in response to stimulation.


Incorporating a Computerized Cognitive Battery Into the Emergency Department Care of Pediatric Mild Traumatic Brain Injuries-Is It Feasible?
The use of computers to test cognitive function acutely after a concussion is becoming increasingly popular, especially after sport-related concussion. Although commonly performed in the community, it is not yet performed routinely in the emergency department (ED), where most injured children present. The challenges of performing computerized cognitive testing (CCT) in a busy ED are considerable. The aim of this study was to evaluate the feasibility of CCT in the pediatric ED after concussion. Children, aged 8 to 18 years with mild traumatic brain injury, presenting to the ED were eligible for this prospective study. Exclusion criteria included the use of drugs, alcohol, and/or physical injury, which could affect CCT performance. A 30- or 15-minute CCT battery was performed. Feasibility measures included environmental factors (space, noise, waiting time), testing factors (time, equipment reliability, personnel), and patient factors (age, injury characteristics). Forty-nine children (28 boys; mean age, 12.6; SD, ± 2.5) participated in the study. All children completed CCT. Mean testing times for the 30- and 15-minute battery were 29.7 and 15.2 minutes, respectively. Noise-cancelling headphones were well tolerated. A shorter CCT was more acceptable to families and was associated with fewer noise disturbances. There was sufficient time to perform testing after triage and before physician assessment in over 90% of children. Computerized cognitive testing is feasible in the ED. We highlight the unique challenges that should be considered before its implementation, including environmental and testing considerations, as well as personnel training.


INSIGHTS INTO AUTOSOMAL DOMINANT STARGARDT-LIKE MACULAR DYSTROPHY THROUGH MULTIMODALITY DIAGNOSTIC IMAGING.
Autosomal dominant Stargardt-like macular dystrophy is a rare juvenile macular dystrophy most commonly because of mutations in ELOVL4 and PROM1 genes. In this study, we review a series of cases of Stargardt-like macular dystrophy and use advanced imaging techniques to describe pathophysiologic manifestations. A retrospective medical record review was performed for five patients from two families with ELOVL4 mutation and one patient with PROM1 mutation including reviewing diagnostic imaging, such as fundus photography, spectral domain optical coherence tomography, fundus autofluorescence, and adaptive optics flood-illuminated photography. All patients had reduced central visual acuity with varying degree of foveal atrophy. In the ELOVL4 group, best-corrected visual acuity ranged from 20/25 to 20/200. Early pathologic changes included thickening of the external limiting membrane and outer nuclear atrophy followed by retinal pigment epithelium loss in later stages. Adaptive optics imaging revealed photoreceptor loss even in early stages with good visual acuity. The PROM1 patient also had similar central vision loss with significant outer nuclear atrophy. In contrast to ELOVL4 mutation, there was more diffuse and patchy retinal pigment epithelium loss throughout the macula. Both ELOVL4- and PROM1-related maculopathies are characterized by progressive photoreceptor atrophy and central vision loss. Using advanced diagnostic imaging, early disease changes and disease progression can be characterized.


Clinical experience with assisted venous drainage cardiopulmonary bypass in elective cardiac reoperations.
Reoperative cardiac surgery is associated with substantial morbidity and mortality due to technical problems at sternal reentry, which can result in laceration of the right ventricle, innominate vein injury, or embolization from patent grafts. To minimize the risk associated with reentry, we adopted the method of assisted venous drainage in the cardiopulmonary bypass circuit with peripheral cannulation for cardiac reoperations. From March 1999 to May 2003, a series of 52 patients (38 males; mean age 48.7 years, range 4 months to 78 years) underwent cardiac reoperations performed with centrifugal pump venous-assisted cardiopulmonary bypass. EuroSCORE was 7.34 +/- 3.9 (range, 4-19). The reoperations were coronary artery bypass graft (25 patients), valve replacement/repair (18 patients), and complex pediatric procedures (11 patients). The studied adverse events were structural damage at reentry, mortality, blood loss, stroke, and hemolysis. Complications at sternotomy were damage to the innominate vein (1 patient) and aorta (1 patient) with blood loss of 625 and 225 mL, respectively. Four patients required intraaortic balloon pump or extracorporeal membrane oxygenation (n = 1) for hemodynamic support on weaning off cardiopulmonary bypass. Three patients died in the postoperative period. Our experience with centrifugal pump-assisted venous drainage in cardiac reoperations has shown excellent results, with reduced risk of damage to vital structures on sternal reentry. In cases in which structural damage did occur, blood loss was minimal.


Regulation of macrophage phagocytosis of apoptotic cells by cAMP.
Regulation of macrophage capacity to remove apoptotic cells may control the balance of apoptotic and necrotic leukocytes at inflamed foci and the extent of leukocyte-mediated tissue damage. Although the molecules involved in the phagocytic process are beginning to be defined, little is known about the underlying regulatory and signaling mechanisms controlling this process. In this paper, we have investigated the effects of treatment of human monocyte-derived macrophages with PGs and other agents that elevate intracellular cAMP on phagocytosis. PGE2 and PGD2 specifically reduced the proportion of macrophages that phagocytosed apoptotic cells. Similar results were obtained with the membrane-permeable cAMP analogues dibutyryl-cAMP and 8-bromo-cAMP but not with the cGMP analogue dibutyryl-GMP. Consistent with the observation that phagocytosis was inhibited by cAMP elevation, treatment of monocyte-derived macrophages with PGE2 resulted in rapid, transient increase in levels of intracellular cAMP. These effects were not due to nonspecific inhibition of monocyte-derived macrophage phagocytosis given that ingestion of Ig-opsonized erythrocytes was unaffected. Elevation of cAMP induced morphologic alterations indicative of changes in the adhesive status of the macrophage, including cell rounding and disassembly of structures that represent points of contact with substrate containing actin and talin. These results strongly suggest that rapid activation of cAMP signaling pathways by inflammatory mediators regulates processes that limit tissue injury and that modulation of cAMP levels represents an additional therapeutic target in the control of resolution of inflammation.


Learning the thyroid examination--a multimodality intervention for internal medicine residents.
Many physicians have inadequate physical diagnosis skills and cannot detect thyroid abnormalities on physical examination. To evaluate a multimodality intervention to improve thyroid examination skills using a prospective controlled trial in first-year residents enrolled in an academic internal medicine program. The intervention group received a 60-minute educational session during which an endocrinologist described anatomical landmarks, thyroid abnormalities, and examination techniques using a slide show, computerized animation, videotape, and live demonstration on a volunteer with goiter. Residents examined a normal and a goitrous thyroid under the observation of a preceptor and received an evidence-based handout on the thyroid examination. The control group received no specific intervention. Examination technique and identification of thyroid abnormalities were blindly assessed in 2 stations of an objective structured clinical examination (OSCE). Of the 19 residents in the intervention group and the 20 in the control group, 6 (32%) and 3 (15%), respectively, observed the neck for thyroid abnormalities (P = 0.3), 17 (90%) and 16 (80%) used proper hand position (P = 0.7), and 13 (68%) and 15 (75%) had the patient swallow while the neck was palpated (P = 0.7). There was a significant difference in the mean scores based on thyroid physical findings during the OSCE between the intervention and control groups (100 vs. 52.5 [maximal possible score = 200], P = 0.047). A 1-hour multimodality learning session furthered the ability of first-year internal medicine residents to detect thyroid abnormalities.


The role of p75NTR in cholinergic basal forebrain structure and function.
The role of the p75 neurotrophin receptor (p75(NTR)) in adult cholinergic basal forebrain (cBF) neurons is unclear due to conflicting results from previous studies and to limitations of existing p75(NTR)-knock-out mouse models. In the present study we used a novel conditional knock-out line (ChAT-cre p75(in/in)) to assess the role of p75(NTR) in the cBF by eliminating p75(NTR) in choline acetyl-transferase-expressing cells. We show that the absence of p75(NTR) results in a lasting increase in cBF cell number, cell size, and cholinergic innervation to the cortex. Analysis of adult ChAT-cre p75(in/in) mice revealed that mutant animals show a similar loss of cBF neurons with age to that observed in wild-type animals, indicating that p75(NTR) does not play a significant role in mediating this age-related decline in cBF neuronal number. However, the increased cholinergic axonal innervation of the cortex, but not the hippocampus, corresponded to alterations in idiothetic but not allothetic navigation. These findings support a role for p75(NTR)-mediated regulation of cholinergic-dependent cognitive function, and suggest that the variability in previous reports of cBF neuron number may stem from limited spatial and temporal control of p75(NTR) expression in existing knock-out models.


Propolis-induced exclusion of colloids: Possible new mechanism of biological action.
Propolis is a natural product originating from life activity of honeybees. It exhibits wide range of biological properties applicable in medicine, the food industry, and cosmetics. Chemically, propolis is a complex and variable mixture with more than 300 identified biologically active components. Propolis's many health-promoting effects are attributed to different biochemical mechanisms, mediated by often-concerted actions of some of its many constituents. Propolis is considered safe and biocompatible. Yet due to its intrinsic complexity, standardization of propolis preparations for medical use as well as prediction of e.g. pathogen-specific interactions becomes a non-trivial task. In this work we demonstrate a new physical mechanism of propolis action, largely independent of specific nuances of propolis chemistry, which may underlie some of its biological actions. We show that propolis-bearing surfaces generate an extensive exclusion zone (EZ) water layer. EZ is an interfacial region of water capable of excluding solutes ranging from ions to microorganisms. Propolis-generated EZ may constitute an effective barrier, physically disabling the approach of various pathogens to the propolis-functionalized surfaces. We suggest possible implications of this new mechanism for propolis-based prevention of respiratory infections.


Bursting neurons signal input slope.
Brief bursts of high-frequency action potentials represent a common firing mode of pyramidal neurons, and there are indications that they represent a special neural code. It is therefore of interest to determine whether there are particular spatial and temporal features of neuronal inputs that trigger bursts. Recent work on pyramidal cells indicates that bursts can be initiated by a specific spatial arrangement of inputs in which there is coincident proximal and distal dendritic excitation (Larkum et al., 1999). Here we have used a computational model of an important class of bursting neurons to investigate whether there are special temporal features of inputs that trigger bursts. We find that when a model pyramidal neuron receives sinusoidally or randomly varying inputs, bursts occur preferentially on the positive slope of the input signal. We further find that the number of spikes per burst can signal the magnitude of the slope in a graded manner. We show how these computations can be understood in terms of the biophysical mechanism of burst generation. There are several examples in the literature suggesting that bursts indeed occur preferentially on positive slopes (Guido et al., 1992; Gabbiani et al., 1996). Our results suggest that this selectivity could be a simple consequence of the biophysics of burst generation. Our observations also raise the possibility that neurons use a burst duration code useful for rapid information transmission. This possibility could be further examined experimentally by looking for correlations between burst duration and stimulus variables.


The M-DC8-positive leukocytes are a subpopulation of the CD14+ CD16+ monocytes.
Recently, a population of M-DC8-positive leukocytes has been described as a new subpopulation of human dendritic cells (DC). In view of the expression of the CD16 antigen on these cells, as well as the finding that DC can arise from blood monocytes, we hypothesized that the expression of M-DC8 is mainly associated with the CD14+ CD16+ phenotype of blood monocytes. Immunofluorescence analysis of whole blood showed that the percentage of M-DC8+ cells is about three times lower than the percentage of CD14+ CD16+ monocytes among all leukocytes (0.32% versus 1.10%). Further, in addition to the expression of CD16, these M-DC8+ cells were also positive for CD14 at low density. Multicolor flow cytometric analysis of whole blood revealed that more than one third of the CD14+ CD16+ monocyte population expressed the M-DC8 antigen (42.3%), and almost all M-DC8+ cells were CD14-CD16-double-positive (87.5%). Finally, the M-DC8 antigen was also expressed on alveolar macrophages from healthy individuals, i.e., cells that are phenotypically and functionally related to the CD14+ CD16+ monocytes. Taken together, the data presented here clearly demonstrate that the M-DC8+ leukocytes are a subpopulation of the CD14+ CD16+ monocytes and may represent DC


Psychological Vulnerability and Problem Gambling: The Mediational Role of Cognitive Distortions.
Despite numerous studies demonstrating the influence of cognitive distortions on gambling problem severity, empirical data regarding the role of psychological vulnerability on the latter is limited. Hence, this study assesses the mediating effect of cognitive distortions between psychological vulnerability (personality and mood), and gambling problem severity. It also verifies whether the relationships between these variables differs according to the preferred gambling activity. The sample is composed of 272 male gamblers [191 poker players; 81 video lottery terminal (VLT) players] aged between 18 and 82 years (M = 35.2). Bootstrap analysis results revealed that cognitive distortions mediate the effect of narcissism on gambling problem severity for both groups. The level of depression for VLT players significantly predicted gambling problem severity, both directly and indirectly via the mediating effect of cognitive distortions. Mediation analyses also indicated that narcissism had an indirect impact on problem gambling through cognitive distortions for both groups. These findings suggest that certain vulnerabilities related to personality and mood may influence cognitive distortion intensity and gambling problem severity. In addition, psychological vulnerabilities could differ based on preferred gambling activity. These results may be useful for prevention policies, identifying high risk gamblers and planning psychological interventions.


Smoking as an Independent Risk Factor for Hepatocellular Carcinoma Due to the α7-Nachr Modulating the JAK2/STAT3 Signaling Axis.
Hepatocellular carcinoma (HCC) is a worldwide health problem. Currently, there is no effective clinical therapeutic strategy for HCC. Smoking is associated with several malignant diseases including cancers. However, the impact of smoking on HCC is still unresolved. Retrospectively reviewed HCC patients diagnosed between 1 January 2010 and 31 December 2015 at Taipei Medical University-Shuang Ho Hospital (Ministry of Health and Welfare). We found that smoking was associated with a poor prognosis, especially recurrence and patient survival after curative surgery using a clinicopathological analysis. Our univariate and multivariate analyses showed that the α7-nicotinic acetylcholine receptor (α7-nAChR) was an oncogene and risk factor for post-resection recurrence. The α7-nAChR was overexpressed in HCC tissues compared to their non-tumor counterparts. Silencing the α7-nAChR reduced the viability of HCC cells, suppressed cellular proliferation, attenuated migration and invasion, and diminished the tumor's sphere-formation ability, with concurrent downregulation of expression levels of the TGR5, p-JAK2, p-STAT3 (Tyr705/Ser727), RhoA, ROCK1, MMP2, and MMP9 proteins. Furthermore, a positive correlation was found between α7-nAChR and JAK2 expressions (p = 0.01) in HCC specimens, as well as their membranous co-localization. Together, we demonstrated that the α7-nAChR may be an independent prognosticator of the progression and prognosis of HCC patients. These findings suggest that the α7-nAChR drives the progression and recurrence of HCC through JAK2/STAT3 signaling and is a novel target for anti-HCC therapy.


Cardiovascular Safety During Treatment With Baricitinib in Rheumatoid Arthritis.
To assess the frequency of cardiovascular and venous thromboembolic events in clinical studies of baricitinib, an oral, selective JAK1 and JAK2 inhibitor approved in more than 50 countries for the treatment of moderately-to-severely active rheumatoid arthritis (RA). Data were pooled from 9 RA studies. Placebo comparison up to 24 weeks included data from 6 studies. Randomized dose comparison between baricitinib doses of 2 mg and 4 mg used data from 4 studies and from the associated long-term extension study. The data analysis set designated "All-bari-RA" included all baricitinib exposures at any dose. Overall, 3,492 RA patients received baricitinib (7,860 patient-years of exposure). No imbalance compared to the placebo group was seen in the incidence of major adverse cardiovascular events (MACE) (incidence rates [IRs] of 0.5 per 100 patient-years for placebo and 0.8 per 100 patient-years for 4 mg baricitinib), arterial thrombotic events (ATE) (IRs of 0.5 per 100 patient-years for placebo and 0.5 per 100 patient-years for 4 mg baricitinib), or congestive heart failure (CHF) broad term (IRs of 4.3 per 100 patient-years for placebo and 2.4 per 100 patient-years for 4 mg baricitinib). Deep vein thrombosis (DVT)/pulmonary embolism (PE) were reported in 0 of 1,070 patients treated with placebo and 6 of 997 patients treated with 4 mg baricitinib during the placebo-controlled period; these events were serious in 2 of 6 patients, while all 6 had risk factors and 1 patient developed DVT/PE after discontinuation of the study drug. In the 2 mg-4 mg-extended data analysis set, IRs of DVT/PE were comparable between the doses across event types (IRs of 0.5 per 100 patient-years in those receiving 2 mg baricitinib and 0.6 per 100 patient-years in those receiving 4 mg baricitinib). In the All-bari-RA data analysis set, the rates were stable over time, with an IR of DVT/PE of 0.5 per 100 patient-years. In RA clinical trials, no association was found between baricitinib treatment and the incidence of MACE, ATE, or CHF. With regard to incidence of DVT/PE, 6 events occurred in patients treated with 4 mg baricitinib, but no cases of DVT/PE were reported in the placebo group. During longer-term evaluation, the incidence of DVT/PE was similar between the baricitinib dose groups, with consistent IR values over time, and this was similar to the rates previously reported in patients with RA.


Modulation of MMP-2 and MMP-9 in Churg-Strauss syndrome respiratory mucosa: potential monitoring parameters.
Churg-Strauss (CSS) syndrome is rare and of unknown etiology. It is associated with vasculitis, blood eosinophilia and granulomatosis, and affects multiple organs and systems at various stages of the disease. Specific diagnostic and monitoring tests are not yet available. This study aims to assess the changes in MMP-2 and MMP-9 along with the histopathological alterations in two cases of CSS, as possible potential diagnostic and monitoring criteria. Two adult male patients were diagnosed with CSS in the otorhinolaryngology clinic in the University of Palermo, based on multiple clinical and histopathologic criteria. Biopsies of respiratory mucosa were taken after the consent of the patients, processed for routine histopathology and immunohistochemistry as well as quantitative polymerase chain reaction (qPCR). Similar biopsies were also taken from a non- CSS patient. The Assessment of MMP-2 and MMP-9 was performed using both immunohistochemistry and qPCR techniques. Histopathological alterations in the respiratory mucosa were consistent with vasculitis and granulomatous tissue formation, in addition to inflammatory cell infiltration with abundance of eosinophils. Immunohistochemistry assay performed on the samples derived from the two CSS patients showed a relative and remarkable increase of both MMP-2 and MMP-9 compared to controls. Such an increase was consistent with the qPCR results which depicted a significant increase between 20 and 30% for both MMP-2 and MMP-9, respectively. Since the secretion of MMPs is an essential step in angiogenesis, could these enzymatic factors be used as parameters to diagnose or monitor the evolution of CSS? The small number of samples analyzed in this study does not allow us to suggest a general statement correlating the increase in expression of MMP-2 and MMP-9 to the appearance or evolution of vasculitis; it is only speculative.


Sexual dysfunction in female cancer survivors.
Cancer survivors face a myriad of long-term effects of their disease, diagnosis, and treatment, and chief among many are problems associated with sexual dysfunction. Yet despite their frequency and the degree of distress they cause patients, sexual dysfunction is not effectively screened for or treated, and this is particularly true in female survivors. Inconsistently performed general sexual health screening at all facets of cancer care and survivorship ultimately translates into missed attempts to identify and treat dysfunction when it does arise. In this paper, we will review the current research and clinical practices addressing sexual dysfunction in female cancer survivors and propose questions in need of future research attention. This article will review the phases of sexual response and how each may be affected by the physical and emotional stress of cancer diagnosis and treatment. We will then discuss existing tools for assessment of sexual function and approaches to their treatment. Finally, we will conclude with advice to health care professionals based on current research and suggest questions for future study.


Calibration processes in desert ant navigation: vector courses and systematic search.
This study investigates the ability of desert ants to adapt their path integration system to an "open-jaw" training paradigm, in which the point of arrival (from the nest) does not coincide with the point of departure (to the nest). Upon departure the ants first run off their home vector and then start a systematic search for the nest. Even if they are subjected to this training-around-a-circuit procedure for more than 50 times in succession, they never adopt straight homeward courses towards the nest. Their path integration vector gets slightly recalibrated (pointing a bit closer to the nest), and their search pattern gets asymmetric (with its search density peak shifted towards the nest), but the bipartite structure of the inbound trajectory invariably remains. These results suggest (1). that the ants cannot learn separate inbound and outbound vectors (i.e. vectors that are not 180 degrees reversals of each other), (2). that the recalibrated vector is dominated by the ant's outbound course, (3). that the recalibration of the vector and the modification of the search geometry are fast and flexible processes occurring whenever the ant experiences a mismatch between the stored and actual states of its path integrator.


Mindfulness meditation for the treatment of chronic low back pain in older adults: a randomized controlled pilot study.
The objectives of this pilot study were to assess the feasibility of recruitment and adherence to an eight-session mindfulness meditation program for community-dwelling older adults with chronic low back pain (CLBP) and to develop initial estimates of treatment effects. It was designed as a randomized, controlled clinical trial. Participants were 37 community-dwelling older adults aged 65 years and older with CLBP of moderate intensity occurring daily or almost every day. Participants were randomized to an 8-week mindfulness-based meditation program or to a wait-list control group. Baseline, 8-week and 3-month follow-up measures of pain, physical function, and quality of life were assessed. Eighty-nine older adults were screened and 37 found to be eligible and randomized within a 6-month period. The mean age of the sample was 74.9 years, 21/37 (57%) of participants were female and 33/37 (89%) were white. At the end of the intervention 30/37 (81%) participants completed 8-week assessments. Average class attendance of the intervention arm was 6.7 out of 8. They meditated an average of 4.3 days a week and the average minutes per day was 31.6. Compared to the control group, the intervention group displayed significant improvement in the Chronic Pain Acceptance Questionnaire Total Score and Activities Engagement subscale (P=.008, P=.004) and SF-36 Physical Function (P=.03). An 8-week mindfulness-based meditation program is feasible for older adults with CLBP. The program may lead to improvement in pain acceptance and physical function.


Mechanistic insights into the cytochrome P450-mediated oxidation and rearrangement of littorine in tropane alkaloid biosynthesis.
During the biosynthesis of certain tropane alkaloids, littorine (1) is rearranged to hyoscyamine (3). Recent evidence indicates that this isomerisation is a two-step process in which the first step is an oxidation/rearrangement to give hyoscyamine aldehyde (2). This step is catalysed by CYP80F1, a cytochrome P450 enzyme, which was recently identified from the plant Hyoscyamus niger; CYP80F1 also catalyses the hydroxylation of littorine at the 3'-position. The mechanisms of the reactions catalysed by CYP80F1 were probed with synthetic deutero and arylfluoro analogues of 1. Measurement of the primary kinetic isotope effects indicates that C3' hydrogen abstraction is the rate-limiting step for the oxidation/rearrangement of natural littorine, and for the 3'-hydroxylation reaction of the unnatural S enantiomer of littorine. The character of the intermediates in the oxidation/rearrangement and hydroxylation reaction was probed with the use of arylfluorinated analogues of (R)-littorine (natural stereoisomer) and (S)-littorine (unnatural stereoisomer) as substrates for CYP80F1. The relative conversions of ortho-, meta- and para-fluorolittorine analogues were used to obtain information on the likely intermediacy of either a benzylic radical or benzylic carbocation intermediate. The data suggest that hydroxylation takes place via a benzylic carbocation intermediate, whereas the product profile arising from rearrangement is more consistent with a benzylic radical intermediate.


Evidence for in vivo interactions between neuropeptide Y-related peptides and sigma receptors in the mouse hippocampal formation.
Recently, it was proposed that neuropeptide Y (NPY) and peptide YY (PYY) could act as endogenous ligands for sigma binding sites, as both NPY and PYY competed with high affinity (nanomolar) for 3H-(+)SKF 10,047 binding sites in rat brain membrane homogenates (Roman et al., 1989). However, various laboratories failed to replicate these in vitro findings. In order to clarify this apparent discrepancy and investigate further possible interactions between NPY and sigma related sites, we evaluated the effects of NPY, PYY and homologs, as well as nonrelated peptides, on in vivo 3H-(+)SKF 10,047 binding parameters in the mouse hippocampal formation. As expected, haloperidol (2 mg/kg), a prototypical sigma receptor ligand, competed for 90% of in vivo hippocampal labeling observed following a peripheral intravenous injection of 3H-(+)SKF 10,047. Intracerebroventricular injections of 300-3000 pmol of either NPY, PYY, NPY2-36, or the Y1 agonist Leu31,Pro34-NPY inhibited significant proportions (17-35%) of haloperidol-sensitive in vivo 3H-(+)SKF 10,047 hippocampal labeling. However, a Y2 receptor agonist, NPY13-36, and nonrelated peptides such as neurotensin and vasoactive intestinal polypeptide, as well as adrenalin, failed to alter in vivo 3H-(+)SKF 10,047 hippocampal binding. It thus appears that NPY, PYY, and a selective Y1 agonist can interact in a concentration-dependent manner, with in vivo 3H-(+)SKF 10,047 labeling in the mouse hippocampal formation. This effect demonstrates selectivity as a Y2 agonist, unrelated peptides, and adrenalin failed to alter in vivo sigma labeling. This in vivo interaction may be relevant to some of the respective biological actions of NPY and sigma-related molecules.


PubMed on Tap: discovering design principles for online information delivery to handheld computers.
Online access to biomedical information from handheld computers will be a valuable adjunct to other popular medical applications if information delivery systems are designed with handheld computers in mind. The goal of this project is to discover design principles to facilitate practitioners' access to online medical information at the point-of-care. A prototype system was developed to serve as a testbed for this research. Using the testbed, an initial evaluation has yielded several user interface design principles. Continued research is expected to discover additional user interface design principles as well as guidelines for results organization and system performance


Microsatellite Alterations With Allelic Loss at 9p24.2 Signify Less-Aggressive Colorectal Cancer Metastasis.
Molecular events that lead to recurrence and/or metastasis after curative treatment of patients with colorectal cancers (CRCs) are poorly understood. Patients with stage II or III primary CRC with elevated microsatellite alterations at selected tetranucleotide repeats and low levels of microsatellite instability (E/L) are more likely to have disease recurrence after treatment. Hypoxia and/or inflammation not only promote metastasis, but also induce elevated microsatellite alterations at selected tetranucleotide repeats by causing deficiency of MSH3 in the cancer cell nucleus. We aimed to identify genetic alterations associated with metastasis of primary colorectal tumors to liver and to determine their effects on survival. We obtained 4 sets of primary colorectal tumors and matched liver metastases from hospitals in Korea and Japan. Intragenic microsatellites with large repeats at 141 loci were examined for frame-shift mutations and/or loss of heterozygosity (LOH) as possible consequences of MSH3 deficiency. Highly altered loci were examined for association with E/L in liver metastases. We analyzed data from 156 of the patients with stage II or III primary colorectal tumors to determine outcomes and whether altered loci were associated with E/L. LOH at several loci at chromosome 9p24.2 (9p24.2-LOH) was associated with E/L in liver metastases (odds ratio = 10.5; 95% confidence interval: 2.69-40.80; P = .0007). We found no significant difference in the frequency of E/L, 9p24.2-LOH, mutations in KRAS or BRAF, or the combination of E/L and 9p24.2-LOH, between primary colorectal tumors and their matched metastases. Patients with stage II or III colorectal tumors with E/L and 9p24.2-LOH had increased survival after CRC recurrence (hazard ratio = 0.25; 95% CI: 0.12-0.50; P = .0001), compared with patients without with E/L and 9p24.2-LOH. E/L with 9p24.2-LOH appeared to be an independent prognostic factor for overall survival of patients with stage III CRC (hazard ratio = 0.06; 95% CI: 0.01-0.57; P = .01). E/L with 9p24-LOH appears to be a biomarker for less aggressive metastasis from stage III primary colorectal tumors.


[Harmonization and toxicological classification of pesticides in 1992 in Brazil and the need to foresee the impacts from the forthcoming introduction of GHS].
In 1992, Brazil modified its toxicological classification criteria for pesticides. This modification aimed to follow the guidelines of the World Health Organization (WHO) hazard classification of pesticides. In 2002, the United Nations adopted the "Globally Harmonized System for the Classification and Labelling of Chemicals" (GHS). As a result, the WHO is reviewing its classification and Brazil will probably follow the example. Therefore it seems timely to estimate the impact of the changes in the criteria adopted in 1992 in the toxicological reclassification and labelling of the commercially available products registered at that time. It is estimated that 58.6% of the total of registered pesticides (74.9% liquid and 31.0% solid formulations) might have been reclassified to less hazardous classes. The hazard warnings on the labels may have caused negative consequences due to misinterpretations by farmers or rural workers. For countries already having a hazard classification system, it is therefore recommendable to estimate the possible impact of the changes before implanting the GHS.


The Healing Effect of Sesame Oil, Camphor and Honey on Second Degree Burn Wounds in Rat.
Many studies were carried out to improve sophisticated dressings to accelerate healing processes and reduce the microbial burden in burn wounds. This study evaluated the healing effect of herbal ointment containing extract of sesame oil, camphor and honey on second degree burn wounds in rats in comparison with daily dressing oil vaseline. Forty rats were randomly assigned to two equal groups. A deep second degree burn was formed on the back of each rat with using a standard burning technique. The burns were dressed daily with herbal ointment containing extract of sesame oil, camphor and honey in group 1, dressing oil vaseline in group 2. The response to treatment was evaluated by digital photography during the treatment on 0, 7, 14, 21, 28 days. Histological scoring was undertaken for scar tissue samples on 0, 7, 14, 21, 28 days. Considerable epithelization in the herbal ointment group vs. the control group over the study period was noted. Neovascularization was significantly higher in herbal ointment treated rats as well. In terms of difference of wound surface area, maximal healing was noticed in herbal ointment extract of sesame oil, camphor and honey group and the minimal repair in the control group. The greatest rate of healing was in the herbal ointment group containing sesame oil, camphor and honey, so the herbal ointment as a suitable substitute for dressing and healing of burn wound injuries is recommended.


Physical optics of the laser-schlieren shock tube technique.
The physical optics of the laser-schlieren technique for the measurement of rate processes in shock waves is examined in detail. The method utilizes the Kirchhoff-Huygens integral with the usual thin lens, paraxial, and Fresnel approximations, all of which are appropriate for the typical laser schlieren experiment. The resolution and sensitivity of the technique are defined for all detector separations, and a reliable method for locating the time origin in the schlieren signal is provided. Diffraction is found to have a significant effect on the shock front generated signal, and geometrical optics treatments of this signal are shown to be inadequate.


Hypoxia potentiates nitric oxide synthesis and transiently increases cytosolic calcium levels in pulmonary artery endothelial cells.
There is indirect, contradictory evidence both for increased and reduced synthesis of the endothelium-derived vasodilator, nitric oxide, in the pulmonary circulation during acute hypoxia. Therefore, we decided to directly measure the effect of acute hypoxia on nitric oxide production by cultured pulmonary endothelium. Because increases in the intracellular free calcium concentration are known to initiate nitric oxide synthesis, we also studied cytosolic calcium levels. We measured the accumulation of the stable nitric oxide metabolite, nitrite, in the fluid used to superfuse the cultured bovine pulmonary artery endothelial cells at an oxygen tension (PO2) of either 20.3 (normoxia) or 4.9 kPa (hypoxia) (152 or 37 mmHg). Intracellular calcium levels were measured with dual-excitation microfluorimetry after loading the cells with the fluorescent calcium indicator, fura 2. Basal NO synthesis, measured as nitrite accumulation over 10 min, was significantly higher under hypoxic than normoxic conditions (8.3 +/- 2.2 versus 4.6 +/- 0.8 nM). Hypoxia transiently increased cytosolic calcium concentration (from 113 +/- 10 to 231 +/- 45 nM). Ryanodine and thapsigargin (which deplete intracellular calcium stores), but not the removal of extracellular calcium, inhibited the hypoxic increase in cytosolic calcium, indicating that it resulted primarily from release of intracellular calcium. Bradykinin-elicited NO synthesis was potentiated by hypoxia. Bradykinin-induced increase in cytosolic calcium was not inhibited by hypoxia. We conclude that hypoxia acutely increases cytosolic calcium levels and basal and bradykinin-stimulated nitric oxide synthesis in pulmonary artery endothelium.


Rous sarcoma virus SRC gene expression on the growth of quail embryo skin fibroblasts and the establishment of permanent cell lines.
Permanent cell lines of Quail embryo fibroblasts appear in cultures of cells infected with a wild type strain of Rous sarcoma virus (SR-RSV) or with its temperature sensitive transformation mutants (ts-T) (NYts68 and PA101) following a three step process. In step one, infected cells grow twice as fast as the control. The second step consists of a crisis during which the cell population is stationary for four to five weeks. Towards the fourth week several foci of cell growth are observed in the flasks. Respreading of the content of these flasks yields permanent lines. This constitutes the third step of the population evolution. In step one the growth rate of the infected cells is the same irrespective of the incubation temperature (36 degrees C or 41 degrees C) whereas the level of the pp60v-src activity is considerably depressed at 41 degrees C for NYts68 and PA101. Foci do not appear at restrictive temperature in the ts infected population and permanent lines are not recovered under that condition. These lines grow ony at 36 degrees C. It can be shown that the virus which they produce is not modified with respect to the temperature sensitivity of the src gene expression since newly infected fibroblasts grow equally well in step one at both 36 degrees C and 41 degrees C, and stop after the same number of generations. This finding suggests that the events which, during the crisis period, lead to the establishment of permanent lines, take place at the cellular level but depend on the activity of the pp60v-src protein for their occurrence or their expression.


Influence of reduced glutathione infusion on glucose metabolism in patients with non-insulin-dependent diabetes mellitus.
To evaluate the relationship between oxidative stress and glucose metabolism, insulin sensitivity and intraerythrocytic reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio were measured in 10 non-insulin-dependent diabetes mellitus (NIDDM) patients and 10 healthy subjects before and after the intravenous administration of GSH. In particular, after baseline insulin sensitivity was assessed by a 2-hour euglycemic hyperinsulinemic clamp, either glutathione (1.35 g x m2 x min(-1)) or placebo (saline) were infused over a period of 1 hour. The same protocol was repeated at a 1-week interval, in cross-over, according to a randomized, single-blind design. In healthy subjects, baseline intraerythrocytic GSH/GSSG ratio (P < .0005) and total glucose uptake (P < .005) were significantly higher than in NIDDM patients. In the same subjects, GSH infusion significantly increased total glucose uptake (from 37.1 +/- 6.7 micromol kg(-1) x min(-1) to 39.5 +/- 7.7 micromol x kg(-1) x min(-1), P < .05), whereas saline infusion was completely ineffective. In addition, the mean intraerythrocytic GSH/GSSG ratio significantly increased after GSH infusion (from 21.0 +/- 0.9 to 24.7 +/- 1.3, P < .05). Similar findings were found in diabetic patients, in whom GSH infusion significantly increased both total glucose uptake (from 25.3 +/- 9.0 micromol x kg(-1) x min(-1) to 31.4 +/- 10.0 micromol x kg(-1) x min(-1), P < .001) and intraerythrocytic GSH/GSSG ratio (from 14.8 +/- 4.1 to 21.7 +/- 6.7, P < .01). Pooling diabetic patients and controls, significant correlations were found between intraerythrocytic GSH/GSSG ratio and total glucose uptake (r = .425, P < .05), as well as between increments of the same variables after GSH infusion (r = .518, P < .05). In conclusion, our data support the hypothesis that abnormal intracellular GSH redox status plays an important role in reducing insulin sensitivity in NIDDM patients. Accordingly, intravenous GSH infusion significantly increased both intraerythrocytic GSH/GSSG ratio and total glucose uptake in the same patients.


Nano-Infrared Imaging of Primary Neurons.
Alzheimer's disease (AD) accounts for about 70% of neurodegenerative diseases and is a cause of cognitive decline and death for one-third of seniors. AD is currently underdiagnosed, and it cannot be effectively prevented. Aggregation of amyloid-β (Aβ) proteins has been linked to the development of AD, and it has been established that, under pathological conditions, Aβ proteins undergo structural changes to form β-sheet structures that are considered neurotoxic. Numerous intensive in vitro studies have provided detailed information about amyloid polymorphs; however, little is known on how amyloid β-sheet-enriched aggregates can cause neurotoxicity in relevant settings. We used scattering-type scanning near-field optical microscopy (s-SNOM) to study amyloid structures at the nanoscale, in individual neurons. Specifically, we show that in well-validated systems, s-SNOM can detect amyloid β-sheet structures with nanometer spatial resolution in individual neurons. This is a proof-of-concept study to demonstrate that s-SNOM can be used to detect Aβ-sheet structures on cell surfaces at the nanoscale. Furthermore, this study is intended to raise neurobiologists' awareness of the potential of s-SNOM as a tool for analyzing amyloid β-sheet structures at the nanoscale in neurons without the need for immunolabeling.


Impact of operating conditions on the removal of endocrine disrupting chemicals by membrane photocatalytic reactor.
This study focuses on the performance of a submerged membrane photocatalytic reactor for the removal of 17beta-oestradiol (E2) in the presence of humic acid (HA). In addition to the impact of operating parameters, such as membrane pore size, ultraviolet (UV) intensity and hydraulic retention time (HRT), the influence of long-term operation was also assessed by advanced characterization of the fouling layer formed on the membrane. The tighter (0.04 microm) hollow fibre polyvinylydene fluoride (PVDF) membrane was found to exhibit not only higher HA removal than the (0.2 microm) module (85% and 75%, respectively), but also greater transmembrane pressure (TMP) values and higher irreversible fouling. Long-term operation conditions have been simulated by conducting an ageing catalyst process and demonstrated a decrease in performance obtained with time. The artificially aged TiO2 resulted in higher TMP values and lower HA removals (about 10-20% decrease) compared with the non-aged catalyst. For E2 removal in the presence of HA, the passive adsorption of the oestrogen onto the organic matter was found to be significant (40% of the E2 adsorbed after I h), demonstrating the importance of the nature of the water matrix for this type of treatment process. An increase in the UV light intensity was observed to favour the E2 elimination, leading to more than 90% removal when using 64 W combined with PVDF membrane and an HRT of 3 h.


Bioaerosol concentrations generated from toilet flushing in a hospital-based patient care setting.
In the United States, 1.7 million immunocompromised patients contract a healthcare-associated infection, annually. These infections increase morbidity, mortality and costs of care. A relatively unexplored route of transmission is the generation of bioaerosols during patient care. Transmission of pathogenic microorganisms may result from inhalation or surface contamination of bioaerosols. The toilet flushing of patient fecal waste may be a source of bioaerosols. To date, no study has investigated bioaerosol concentrations from flushing fecal wastes during patient care. Particle and bioaerosol concentrations were measured in hospital bathrooms across three sampling conditions; no waste no flush, no waste with flush, and fecal waste with flush. Particle and bioaerosol concentrations were measured with a particle counter bioaerosol sampler both before after a toilet flushing event at distances of 0.15, 0.5, and 1 m from the toilet for 5, 10, 15 min. Particle concentrations measured before and after the flush were found to be significantly different (0.3-10 μm). Bioaerosol concentrations when flushing fecal waste were found to be significantly greater than background concentrations (p-value = 0.005). However, the bioaerosol concentrations were not different across time (p-value = 0.977) or distance (p-value = 0.911) from the toilet, suggesting that aerosols generated may remain for longer than 30 min post flush. Toilets produce aerosol particles when flushed, with the majority of the particles being 0.3 μm in diameter. The particles aerosolized include microorganisms remaining from previous use or from fecal wastes. Differences in bioaerosol concentrations across conditions also suggest that toilet flushing is a source of bioaerosols that may result in transmission of pathogenic microorganisms. This study is the first to quantify particles and bioaerosols produced from flushing a hospital toilet during routine patient care. Future studies are needed targeting pathogens associated with gastrointestinal illness and evaluating aerosol exposure reduction interventions.


CO Methanation for Synthetic Natural Gas Production.
Energy from woody biomass could supplement renewable energy production towards the replacement of fossil fuels. A multi-stage process involving gasification of wood and then catalytic transformation of the producer gas to synthetic natural gas (SNG) represents progress in this direction. SNG can be transported and distributed through the existing pipeline grid, which is advantageous from an economical point of view. Therefore, CO methanation is attracting a great deal of attention and much research effort is focusing on the understanding of the process steps and its further development. This short review summarizes recent efforts at Paul Scherrer Institute on the understanding of the reaction mechanism, the catalyst deactivation, and the development of catalytic materials with benign properties for CO methanation.


Feedback control for microring weight banks.
Microring weight banks present novel opportunities for reconfigurable, high-performance analog signal processing in photonics. Controlling microring filter response is a challenge due to fabrication variations and thermal sensitivity. Prior work showed continuous weight control of multiple wavelength-division multiplexed signals in a bank of microrings based on calibration and feedforward control. Other prior work has shown resonance locking based on feedback control by monitoring photoabsorption-induced changes in resistance across in-ring photoconductive heaters. In this work, we demonstrate continuous, multi-channel control of a microring weight bank with an effective 5.1 bits of accuracy on 2Gbps signals. Unlike resonance locking, the approach relies on an estimate of filter transmission versus photo-induced resistance changes. We introduce an estimate still capable of providing 4.2 bits of accuracy without any direct transmission measurements. Furthermore, we present a detailed characterization of this response for different values of carrier wavelength offset and power. Feedback weight control renders tractable the weight control problem in reconfigurable analog photonic networks.


Digoxin in atrial fibrillation: report from the Stockholm Cohort study of Atrial Fibrillation (SCAF).
Recent studies of patients with heart failure and of patients receiving intensive care indicate that digoxin may increase mortality if the patient has atrial fibrillation (AF). Objective To study which patients receive digoxin treatment for AF and what the prognosis is for patients given this treatment. 2824 patients with AF were studied prospectively for a mean of 4.6 years. Information about medication was obtained from the local hospital registry. Information about diagnoses, hospitalisations and deaths was obtained from national registries. Propensity score matching and Cox regression was used to account for confounding. Factors associated with digoxin use were permanent AF (hazard ratio (HR) = 3.2, confidence interval (CI) 2.7 to 3.9), absence of pacemaker (HR = 2.3, CI 1.6 to 3.2), history of heart failure (HR = 2.0, CI 1.7 to 2.5), treatment in an internal medicine ward rather than a cardiology ward (HR = 1.6, CI 1.3 to 2.0), female sex (HR = 1.6, CI 1.3 to 1.9) and age >or=80 years (HR = 1.4, CI 1.1 to 1.7). More patients with than without digoxin died (51% vs 31%, p<0.001). After adjustment for covariates, however, no disadvantages related to digoxin use could be found for all-cause mortality, myocardial infarction, ischaemic stroke, time to readmission to hospital or days at hospital/year at risk. The only end point significantly associated with digoxin use was pacemaker implantations, which were more common in digoxin-treated patients (HR = 2.0, CI 1.2 to 3.4). Digoxin is mainly given to an elderly and frailer subset of patients with AF and is thus associated with an increased mortality. When differences in patient characteristics are accounted for digoxin use seems to have a neutral effect on mortality and major cardiovascular events in patients with AF.


Stinging nettle and neem enhance antibody response to local killed and imported live infectious bursal disease vaccines in indigenous chicken in Kenya.
Immune responses are critical for protection of chickens from infectious bursal disease (IBD). In this study, the antibody response-enhancing effect of drinking water supplementation of 1% stinging nettle and neem on different IBD vaccines and vaccination regimes was evaluated, using 36 (n = 36) specific antibody negative indigenous chicks. The birds were allocated into 3 groups as follows: 1A-C, 2A-C, and 3A-B, while group 3C acted as the unvaccinated non-supplemented control. A local inactivated K1 and imported live attenuated D78 IBD vaccines were given to groups 1A-C and 3A-B at 14 and 28 d of age, respectively. A combination of K1 and D78 vaccines was given 30 d apart to groups 2A and 2B (D78 at 14 and 21 d and K1 at 44 d of age) and on the same d to group 2C at 14 and 28 d of age. Stinging nettle was given in water to groups 1B, 2B, and 2C, and neem to groups 1C, 2A, and 3B. Birds were bled weekly and immune responses monitored using indirect ELISA. Both neem and stinging nettle had antibody response-enhancing effects in groups 1B and 1C, receiving the local inactivated K1 vaccine. There were significant differences (P < 0.05) in antibody titers between groups 1A and 2C. Stinging nettle induced earlier onset of high antibody responses in group 2C and persistent titers (>3.8 log10) from the third week in group 2B. Imported live D78 vaccine induced higher antibody titers compared to the local inactivated K1 vaccine. Groups 2B and 2C receiving a combination of the local K1 and imported live attenuated D78 vaccines had the highest antibody titers. Adoption of stinging nettle supplementation and a prime-boost program involving use of a local virus isolates-derived vaccine is recommended.


Frataxin deficiency alters heme pathway transcripts and decreases mitochondrial heme metabolites in mammalian cells.
Deficiency of the frataxin mRNA alters the transcriptome, triggering neuro- and cardiodegeneration in Friedreich's ataxia. We microarrayed murine frataxin-deficient heart tissue, liver tissue and cardiocytes and observed a transcript down-regulation to up-regulation ratio of nearly 2:1 with a mitochondrial localization of transcriptional changes. Combining all mouse and human microarray data for frataxin-deficient cells and tissues, the most consistently decreased transcripts were mitochondrial coproporphyrinogen oxidase (CPOX) of the heme pathway and mature T-cell proliferation 1, a homolog of yeast COX23, which is thought to function as a mitochondrial metallochaperone. Quantitative RT-PCR studies confirmed the significant down-regulation of Isu1, CPOX and ferrochelatase at 10 weeks in mouse hearts. We observed that mutant cells were resistant to aminolevulinate-dependent toxicity, as expected if the heme pathway was inhibited. Consistent with this, we observed increased cellular protoporphyrin IX levels, reduced mitochondrial heme a and heme c levels and reduced activity of cytochrome oxidase, suggesting a defect between protoporphyrin IX and heme a. Fe-chelatase activities were similar in mutants and controls, whereas Zn-chelatase activities were slightly elevated in mutants, supporting the idea of an altered metal-specificity of ferrochelatase. These results suggest that frataxin deficiency causes defects late in the heme pathway. As ataxic symptoms occur in other diseases of heme deficiency, the heme defect we observe in frataxin-deficient cells could be primary to the pathophysiological process.


Good Early Results Obtained with a Guided-Motion Implant for Total Knee Arthroplasty: A Consecutive Case Series.
Previous studies have shown a high incidence of complications with a bi-cruciate stabilized (BCS) guided-motion total knee arthroplasty (TKA) design, which led to recent modifications of the design by the manufacturer. The current study was undertaken to assess whether the use of this TKA system with an extension-first surgical technique is associated with a similar rate of short-term adverse outcome as reported in literature. This retrospective study enrolled 257 consecutive patients (257 knees) undergoing TKA for osteoarthritis of the knee, with the first 153 receiving cemented Journey BCS I implants and the remaining 104 receiving cemented Journey BCS II implants when these became available. Mean follow-up time for the cohort was 24.5 ± 7.8 months (range, 12 - 36 months). There were no cases of stiffness. Incidence of iliotibial friction syndrome was considered low: three (2.0%) knees in the BCS I group and two (1.9%) in the BCS II group (p = 0.676). Five (2.5%) knees presented with mild instability in midflexion, three (2.0%) in the BCS I group and two (1.9%) in the BCS II group (p = 0.676). One patient with a BCS I implant required reoperation for aseptic loosening 23 months postoperatively. At one-year follow-up, there were no clinically relevant differences in any of the clinical outcomes. When used in combination with an extension-first surgical technique, good early functional results with an acceptable rate of complications were obtained with both the original and the updated Journey BCS knee implant.


Helical membrane protein conformations and their environment.
Evidence that membrane proteins respond conformationally and functionally to their environment is growing. Structural models, by necessity, have been characterized in preparations where the protein has been removed from its native environment. Different structural methods have used various membrane mimetics that have recently included lipid bilayers as a more native-like environment. Structural tools applied to lipid bilayer-embedded integral proteins are informing us about important generic characteristics of how membrane proteins respond to the lipid environment as compared with their response to other nonlipid environments. Here, we review the current status of the field, with specific reference to observations of some well-studied α-helical membrane proteins, as a starting point to aid the development of possible generic principles for model refinement.


The chromosome-scale reference genome of safflower (Carthamus tinctorius) provides insights into linoleic acid and flavonoid biosynthesis.
Safflower (Carthamus tinctorius L.), a member of the Asteraceae, is a popular crop due to its high linoleic acid (LA) and flavonoid (such as hydroxysafflor yellow A) contents. Here, we report the first high-quality genome assembly (contig N50 of 21.23 Mb) for the 12 pseudochromosomes of safflower using single-molecule real-time sequencing, Hi-C mapping technologies and a genetic linkage map. Phyloge nomic analysis showed that safflower diverged from artichoke (Cynara cardunculus) and sunflower (Helianthus annuus) approximately 30.7 and 60.5 million years ago, respectively. Comparative genomic analyses revealed that uniquely expanded gene families in safflower were enriched for those predicted to be involved in lipid metabolism and transport and abscisic acid signalling. Notably, the fatty acid desaturase 2 (FAD2) and chalcone synthase (CHS) families, which function in the LA and flavonoid biosynthesis pathways, respectively, were expanded via tandem duplications in safflower. CarFAD2-12 was specifically expressed in seeds and was vital for high-LA content in seeds, while tandemly duplicated CarFAD2 genes were up-regulated in ovaries compared to CarFAD2-12, which indicates regulatory divergence of FAD2 in seeds and ovaries. CarCHS1, CarCHS4 and tandem-duplicated CarCHS5˜CarCHS6, which were up-regulated compared to other CarCHS members at early stages, contribute to the accumulation of major flavonoids in flowers. In addition, our data reveal multiple alternative splicing events in gene families related to fatty acid and flavonoid biosynthesis. Together, these results provide a high-quality reference genome and evolutionary insights into the molecular basis of fatty acid and flavonoid biosynthesis in safflower.


Role of the long slender to short stumpy transition in the life cycle of the african trypanosomes.
It is shown using mouse models that the African trypanosomes exert a significant drain upon their host's carbohydrate (energy) resources; and that the higher the parasitemia, the greater the energy demand. It is, therefore, hypothesized that the long slender (LS) to short stumpy (SS) transition evolved, in part, to help control the parasitemia and to increase host survival time. It is also suggested that the SS population is heterogeneous. One part of the population is tsetse infective, while a second older SS population is undergoing apoptotic-like events, which leads to their cell death and their stimulation of the host's immune response. This immune stimulation by the old dying SS forms would eliminate the major LS and SS variant antigen population, and produce the chronic relapsing infection. It is concluded that the SS stages during the apoptosis-like process are acting altruistically. They give their lives to insure the long-term survival of the host, and to insure renewed growth of the minor LS variants and new infective SS forms. This process is predicted to increase the probability for the successful transmission of the trypanosomes to a new host.


Expression of a calmodulin-dependent phosphodiesterase isoform (PDE1B1) correlates with brain regions having extensive dopaminergic innervation.
Cyclic nucleotide-dependent protein phosphorylation plays a central role in neuronal signal transduction. Neurotransmitter-elicited increases in cAMP/cGMP brought about by activation of adenylyl and guanylyl cyclases are downregulated by multiple phosphodiesterase (PDE) enzymes. In brain, the calmodulin (CaM)-dependent isozymes are the major degradative activities and represent a unique point of intersection between the cyclic nucleotide- and calcium (Ca2+)-mediated second messenger systems. Here we describe the distribution of the PDE1B1 (63 kDa) CaM-dependent PDE in mouse brain. An anti-peptide antiserum to this isoform immunoprecipitated approximately 30-40% of cytosolic PDE activity, whereas antiserum to PDE1A2 (61 kDa isoform) removed 60-70%, demonstrating that these isoforms are the major CaM-dependent PDEs in brain. Quantification of PDE1B1 immunoreactivity on immunoblots indicated that striatum contains 3-17-fold higher levels of PDE1B1 than other brain regions, with lowest immunoreactivity in cerebellum. In situ hybridization demonstrated high levels of PDE1B1 mRNA in the caudate-putamen, nucleus accumbens, and olfactory tubercle. Moderate mRNA levels were observed in dentate gyrus, cerebral cortex, medial thalamic nuclei, and brainstem, whereas negligible mRNA was detectable in the globus pallidus, islands of Calleja, substantia nigra, and ventral tegmental area. Immunocytochemistry confirmed that the majority of PDE1B1 protein was localized to the caudate-putamen, nucleus accumbens, and olfactory tubercle. Within the caudate-putamen, PDE1B1 immunoreactivity was ubiquitous, while PDE1A2 immunostaining was restricted to a minor subset of striatal neurons. The expression of PDE1B1 protein and mRNA correlate strongly with areas of the brain that are richest in dopaminergic innervation; indeed, there are strikingly similar distributions for PDE1B1 and D1 dopamine receptor mRNAs. Since D1 receptor binding activates adenylyl cyclase, and striatal neurons lack CaM-sensitive forms of cyclase, the high amount of this PDE implies an important physiological role for Ca(2+)-regulated attenuation of cAMP-dependent signaling pathways following dopaminergic stimulation.


Functional divergence of a global regulatory complex governing fungal filamentation.
Morphogenetic transitions are prevalent in the fungal kingdom. For a leading human fungal pathogen, Candida albicans, the capacity to transition between yeast and filaments is key for virulence. For the model yeast Saccharomyces cerevisiae, filamentation enables nutrient acquisition. A recent functional genomic screen in S. cerevisiae identified Mfg1 as a regulator of morphogenesis that acts in complex with Flo8 and Mss11 to mediate transcriptional responses crucial for filamentation. In C. albicans, Mfg1 also interacts physically with Flo8 and Mss11 and is critical for filamentation in response to diverse cues, but the mechanisms through which it regulates morphogenesis remained elusive. Here, we explored the consequences of perturbation of Mfg1, Flo8, and Mss11 on C. albicans morphogenesis, and identified functional divergence of complex members. We observed that C. albicans Mss11 was dispensable for filamentation, and that overexpression of FLO8 caused constitutive filamentation even in the absence of Mfg1. Harnessing transcriptional profiling and chromatin immunoprecipitation coupled to microarray analysis, we identified divergence between transcriptional targets of Flo8 and Mfg1 in C. albicans. We also established that Flo8 and Mfg1 cooperatively bind to promoters of key regulators of filamentation, including TEC1, for which overexpression was sufficient to restore filamentation in the absence of Flo8 or Mfg1. To further explore the circuitry through which Mfg1 regulates morphogenesis, we employed a novel strategy to select for mutations that restore filamentation in the absence of Mfg1. Whole genome sequencing of filamentation-competent mutants revealed chromosome 6 amplification as a conserved adaptive mechanism. A key determinant of the chromosome 6 amplification is FLO8, as deletion of one allele blocked morphogenesis, and chromosome 6 was not amplified in evolved lineages for which FLO8 was re-located to a different chromosome. Thus, this work highlights rewiring of key morphogenetic regulators over evolutionary time and aneuploidy as an adaptive mechanism driving fungal morphogenesis.


Mortality among patients with psychiatric illness. A ten-year case register study in an area with a community-based system of care.
Most studies which showed an excess mortality in psychiatric patients have been conducted on hospitalised samples. This was a case register study. All South Verona patients with an ICD diagnosis who had psychiatric contacts with specialist services in 1982-1991 were included. Mortality was studied in relation to sex, age, diagnosis, pattern of care and interval from registration. Standardised Mortality Rates (SMRs) and Poisson regression analysis were calculated. The overall SMR was 1.63 (95% Cl = 1.5-1.8), which is the lowest value reported so far. Mortality was higher among men (SMR = 2.24; 95% Cl = 1.9-2.6), among patients who were admitted to hospital (SMR = 2.23; 95% Cl = 1.9-2.6), among younger age groups (SMR = 8.82; 95% Cl = 4.9-14.6) and in the first year after registration (SMR = 2.32; 95% CL = 1.8-2.9). Higher mortality was found in patients with a diagnosis of alcohol and drug dependence (SMR = 3.87; 95% Cl = 3.0-4.9). The SMR for suicide was 17.41. Using a Poisson regression model, diagnosis, pattern of care and interval from registration were all found to be significantly associated with mortality. When all these variables were entered together in the model, each maintained its predictive role. The overall mortality of psychiatric patients treated in a community-based system of care was higher than expected, but lower than the mortality reported in other psychiatric settings. The highest mortality risk was found in the first year after registration.


Natural processing sites for human cathepsin E and cathepsin D in tetanus toxin: implications for T cell epitope generation.
Cathepsin E is an aspartic proteinase that has been implicated frequently in Ag processing for presentation on class II MHC molecules, but no information exists on its cleavage specificity within Ags in relation to known T cell epitopes. We have analyzed the processing by cathepsin E of a large C-terminal domain of tetanus toxin (residues 872-1315), and we have compared the processing products with those liberated by cathepsin D, a related aspartic proteinase also thought to be involved in class II MHC-restricted Ag processing. Processing products were analyzed by N-terminal Edman degradation and mass spectrometry following reverse-phase HPLC separation of peptides. A total of 28 cleavage sites was identified, 11 of which were recognized by both cathepsins E and D. Most, although not all, sites were between pairs of hydrophobic residues and were located within the 200-amino-acid C terminal region known to contain several human T cell epitopes. Previously described T cell epitopes, for example, between residues 1273 and 1284, were flanked by cathepsin E and D cleavage sites. These data are consistent with an important role for cathepsins E and/or D in Ag processing in the human immune system.


Conceptualizing and communicating management effects on forest water quality.
We present a framework for evaluating and communicating effects of human activity on water quality in managed forests. The framework is based on the following processes: atmospheric deposition, weathering, accumulation, recirculation and flux. Impairments to water quality are characterized in terms of their extent, longevity and frequency. Impacts are communicated using a "traffic lights" metaphor for characterizing severity of water quality impairments arising from forestry and other anthropogenic pressures. The most serious impairments to water quality in managed boreal forests include (i) forestry activities causing excessive sediment mobilization and extirpation of aquatic species and (ii) other anthropogenic pressures caused by long-range transport of mercury and acidifying pollutants. The framework and tool presented here can help evaluate, summarize and communicate the most important issues in circumstances where land management and other anthropogenic pressures combine to impair water quality and may also assist in implementing the "polluter pays" principle.