Probe technologies for clean sampling and measurement of subglacial lakes.
It is 4 years since the subglacial lake community published its plans for accessing, sampling, measuring and studying the pristine, and hitherto enigmatic and very different, Antarctic subglacial lakes, Vostok, Whillans and Ellsworth. This paper summarizes the contrasting probe technologies designed for each of these subglacial environments and briefly updates how these designs changed or were used differently when compared to previously published plans. A detailed update on the final engineering design and technical aspects of the probe for Subglacial Lake Ellsworth is presented. This probe is designed for clean access, is negatively buoyant (350 kg), 5.2 m long, 200 mm in diameter, approximately cylindrical and consists of five major units: (i) an upper power and communications unit attached to an optical and electrical conducting tether, (ii)-(iv) three water and particle samplers, and (v) a sensors, imaging and instrumentation pack tipped with a miniature sediment corer. To date, only in Subglacial Lake Whillans have instruments been successfully deployed. Probe technologies for Subglacial Lake Vostok (2014/15) and Lake Ellsworth (2012/13) were not deployed for technical reasons, in the case of Lake Ellsworth because hot-water drilling was unable to access the lake during the field season window. Lessons learned and opportunities for probe technologies in future subglacial access missions are discussed.


Convergent evolution of vocal cooperation without convergent evolution of brain size.
One pragmatic underlying successful vocal communication is the ability to take turns. Taking turns - a form of cooperation - facilitates the transmission of signals by reducing the amount of their overlap. This allows vocalizations to be better heard. Until recently, non-human primates were not thought of as particularly cooperative, especially in the vocal domain. We recently demonstrated that common marmosets (Callithrix jacchus), a small New World primate species, take turns when they exchange vocalizations with both related and unrelated conspecifics. As the common marmoset is distantly related to humans (and there is no documented evidence that Old World primates exhibit vocal turn taking), we argue that this ability arose as an instance of convergent evolution, and is part of a suite of prosocial behavioral tendencies. Such behaviors seem to be, at least in part, the outcome of the cooperative breeding strategy adopted by both humans and marmosets. Importantly, this suite of shared behaviors occurs without correspondence in encephalization. Marmoset vocal turn taking demonstrates that a large brain size and complex cognitive machinery is not needed for vocal cooperation to occur. Consistent with this idea, the temporal structure of marmoset vocal exchanges can be described in terms of coupled oscillator dynamics, similar to quantitative descriptions of human conversations. We propose a simple neural circuit mechanism that may account for these dynamics and, at its core, involves vocalization-induced reductions of arousal. Such a mechanism may underlie the evolution of vocal turn taking in both marmoset monkeys and humans.


Cross-linking mass spectrometry identifies new interfaces of Augmin required to localise the γ-tubulin ring complex to the mitotic spindle.
The hetero-octameric protein complex, Augmin, recruits γ-Tubulin ring complex (γ-TuRC) to pre-existing microtubules (MTs) to generate branched MTs during mitosis, facilitating robust spindle assembly. However, despite a recent partial reconstitution of the human Augmin complex in vitro, the molecular basis of this recruitment remains unclear. Here, we used immuno-affinity purification of in vivo Augmin from Drosophila and cross-linking/mass spectrometry to identify distance restraints between residues within the eight Augmin subunits in the absence of any other structural information. The results allowed us to predict potential interfaces between Augmin and γ-TuRC. We tested these predictions biochemically and in the Drosophila embryo, demonstrating that specific regions of the Augmin subunits, Dgt3, Dgt5 and Dgt6 all directly bind the γ-TuRC protein, Dgp71WD, and are required for the accumulation of γ-TuRC, but not Augmin, to the mitotic spindle. This study therefore substantially increases our understanding of the molecular mechanisms underpinning MT-dependent MT nucleation.


Amphetamine promotes task-dependent recovery following focal cortical ischaemic lesions in the rat.
This study investigated the effect of amphetamine (AMP) on skilled forelimb use following focal cortical ischaemic lesions in the rat. Unilateral lesions were produced by a novel method of intracortical microinjection of endothelin-1 (ET-1), intended to principally target the forelimb representation zone in primary motor-primary somatosensory cortex. Lesions were placed in the hemisphere contralateral to the preferred limb and produced deficits in skilled forelimb use on two tasks: the paw reach (PR) test and the foot fault (FF) test. Beginning on post-lesion day (D) 2, animals received injections of 2 mg/kg AMP and were injected every third day until D26. Animals were tested both during, and 24 h after, AMP administration. AMP facilitated recovery of skilled forelimb use on the PR test when assessed during drug-free test sessions. No such effect was seen on the FF test. These results demonstrate that sub-acute administration of AMP following a unilateral focal ischaemic lesion of FL can facilitate task-dependent recovery of skilled forelimb use in the rat. They also demonstrate that different behavioural tasks measuring superficially similar behavioural outputs may show different sensitivities to such drug effects.


Is the Rotavirus Vaccine Really Associated with a Decreased Risk of Developing Celiac and Other Autoimmune Diseases?
This review examines the risk of developing celiac disease (CD) and other autoimmune diseases in individuals receiving the rotavirus (RV) vaccine compared to the normal population. Celiac disease is a malabsorptive, chronic, immune-mediated enteropathy involving the small intestine. The pathogenesis of CD is multifactorial, and mucosal immunity plays an important role in its development. Low mucosal IgA levels significantly increase the risk of developing the disease. Rotavirus is an infectious agent that causes diarrhea, particularly in children aged 0-24 months, and is frequently involved in diarrhea-related deaths in these children. An oral vaccine against RV has been developed. While it is effective on RV infection, it also contributes to increasing mucosal immunity. Studies have indicated that individuals immunized with the RV vaccine are at lower risk of developing CD than unvaccinated individuals. In addition, the mean age for developing CD autoimmunity may be higher in the vaccinated group than in controls receiving placebo. Additional studies that include children immunized with different RV vaccines and unvaccinated children would provide more meaningful results. Although current data suggest a possible association of RV vaccination with a reduced risk of developing CD and other autoimmune diseases, this remains an unanswered question that merits greater international investigation.


"Who's been a good dog?" - Owner perceptions and motivations for treat giving.
Complex relationships commonly exist between owners and their companion animals, particularly around feeding behaviour with an owner's affection or love for their animal most pronounced through the provision of food. It is notable that the pet food market is experiencing strong year-on-year growth in sales of dog and cat treats. Recognising the impact of treat giving in pet nutrition, the objective of the study was to investigate owner attitudes and motivations towards feeding treats (shop bought and other) to their dogs. A researcher-mediated questionnaire consisting of both quantitative and qualitative questions was used to interview dog owners (n=280) at two locations: an out-of-town retail park and a country park in the East Midlands. Owners almost unanimously viewed the word 'treat' within a nutritional context, as opposed to a new toy or other pleasure. The majority (96%) of owners interviewed reported feeding treats to their dog, with 69% feeding shop-bought treats on a daily basis. A wide range of treats was reportedly given by owners and the majority of owners interviewed fed multiple treat types. No association was found between owner age and frequency of shop-bought treats fed (P=0.659) nor between owner age and frequency of food given to the dog from the owner's plate (P=0.083). A wide range of foods which would not be considered balanced for the animal's nutritional requirements was viewed as a treat by some dog owners. A range of positive and negative views around the feeding of treats were expressed by dog owners, with some citing beneficial effects while others were clearly aware of the association between treat feeding and potential weight gain/obesity. Owner views included themes around positive reinforcement and responsibility but also reflected relational aspects of the human-animal bond. The results of the study show that treat giving is commonplace in feeding regimes and that treats are embedded in the feeding behaviour of many dog owners. However, the different views expressed around the motivations for, and feeding of, dog treats, reinforce the need to better understand owner psychology linked to this area, and the role this may play in the growing pet obesity epidemic.


Presentation and Outcomes of Localized Immunoglobulin Light Chain Amyloidosis: The Mayo Clinic Experience.
To describe treatment types, outcomes, and relapse patterns in patients with localized immunoglobulin light chain amyloidosis (ALL). We included all patients with ALL seen at Mayo Clinic in Rochester, Minnesota, from January 1, 1968, through June 30, 2014. The diagnosis of ALL was predicated on the presence of a Congo red-positive biopsy specimen and negative serum and urine immunofixation. Treatment response categories were response, stability, and progression. Localized and systemic progressions were defined as progression of disease at the site of origin or appearance of clonal plasma cells in a bone marrow biopsy sample, respectively. Of 5551 patients with AL, 413 (7%) had ALL. The most common site involved was urothelial tissue (n=85, 21%), followed by the larynx (n=57, 14%). Coexisting autoimmune diseases were reported in 7% of patients (n=28). The most common first-line treatment was excision of the amyloid deposits (61%), followed by observation or supportive care (28%). When considering symptomatic patients only (n=284), 205 (72%) improved, 23 (8%) had stable disease, and 55 (19%) could not be evaluated for response. Ten-year survival was 78% and was not different from that of the general population. There were no systemic progressions, but 17% of patients (n=72) had localized progression. Localized AL is associated with a relatively distinct pattern of organ involvement. The initial laboratory evaluation to exclude systemic disease could be limited to serum and urine immunofixation in most patients. Recurrence after first-line therapy is common, but long-term outcomes are excellent.


Kafka, paranoic doubles and the brain: hypnagogic vs. hyper-reflexive models of disrupted self in neuropsychiatric disorders and anomalous conscious states.
Kafka's writings are frequently interpreted as representing the historical period of modernism in which he was writing. Little attention has been paid, however, to the possibility that his writings may reflect neural mechanisms in the processing of self during hypnagogic (i.e., between waking and sleep) states. Kafka suffered from dream-like, hypnagogic hallucinations during a sleep-deprived state while writing. This paper discusses reasons (phenomenological and neurobiological) why the self projects an imaginary double (autoscopy) in its spontaneous hallucinations and how Kafka's writings help to elucidate the underlying cognitive and neural mechanisms. I further discuss how the proposed mechanisms may be relevant to understanding paranoid delusions in schizophrenia. Literature documents and records cognitive and neural processes of self with an intimacy that may be otherwise unavailable to neuroscience. To elucidate this approach, I contrast it with the apparently popularizing view that the symptoms of schizophrenia result from what has been called an operative (i.e., pre-reflective) hyper-reflexivity. The latter approach claims that pre-reflective self-awareness (diminished in schizophrenia) pervades all conscious experience (however, in a manner that remains unverifiable for both phenomenological and experimental methods). This contribution argues the opposite: the "self" informs our hypnagogic imagery precisely to the extent that we are not self-aware.


Cannabinoids enhance subsecond dopamine release in the nucleus accumbens of awake rats.
Dopaminergic neurotransmission has been highly implicated in the reinforcing properties of many substances of abuse, including marijuana. Cannabinoids activate ventral tegmental area dopaminergic neurons, the main ascending projections of the mesocorticolimbic dopamine system, and change their spiking pattern by increasing the number of impulses in a burst and elevating the frequency of bursts. Although they also increase time-averaged striatal dopamine levels for extended periods of time, little is known about the temporal structure of this change. To elucidate this, fast-scan cyclic voltammetry was used to monitor extracellular dopamine in the nucleus accumbens of freely moving rats with subsecond timescale resolution. Intravenous administration of the central cannabinoid (CB1) receptor agonist, R(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-(1-naphthalenyl) methanone mesylate, dose-dependently produced catalepsy, decreased locomotion, and reduced the amplitude of electrically evoked dopamine release while markedly increasing the frequency of detected (nonstimulated) dopamine concentration transients. The CB1 receptor antagonist [N-piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methylpyrazole-3-carboxamide] reversed and prevented all agonist-induced effects but did not show effects on dopamine release when injected alone. These data demonstrate that doses of a cannabinoid agonist known to increase burst firing produce ongoing fluctuations in extracellular dopamine on a previously unrecognized temporal scale in the nucleus accumbens.


The protective effect of diosmin on hepatic ischemia reperfusion injury: an experimental study.
Liver ischemia reperfusion injury (IRI) is an important pathologic process leading to bodily systemic effects and liver injury. Our study aimed to investigate the protective effects of diosmin, a phlebotrophic drug with antioxidant and anti-inflammatory effects, in a liver IRI model. Forty rats were divided into 4 groups. Sham group, control group (ischemia-reperfusion), intraoperative treatment group, and preoperative treatment group. Ischemia reperfusion model was formed by clamping hepatic pedicle for a 60 minute of ischemia followed by liver reperfusion for another 90 minutes. Superoxide dismutase (SOD) and catalase (CAT) were measured as antioaxidant enzymes in the liver tissues, and malondialdehyde (MDA) as oxidative stress marker, xanthine oxidase (XO) as an oxidant enzyme and glutathione peroxidase (GSH-Px) as antioaxidant enzyme were measured in the liver tissues and the plasma samples. Hepatic function tests were lower in treatment groups than control group (p<0.001 for ALT and AST). Plasma XO and MDA levels were lower in treatment groups than control group, but plasma GSH-Px levels were higher (p<0.05 for all). Tissue MDA levels were lower in treatment groups than control group, but tissue GSH-Px, SOD, CAT and XO levels were higher (p<0.05 for MDA and p<0.001 for others). Samples in control group histopathologically showed morphologic abnormalities specific to ischemia reperfusion. It has been found that both preoperative and intraoperative diosmin treatment decreases cellular damage and protects cells from toxic effects in liver IRI. As a conclusion, diosmin may be used as a protective agent against IRI in elective and emergent liver surgical operations.


A comparison of subadult skeletal and dental development based on living and deceased samples.
A fundamental assumption in biological anthropology is that living individuals will present with different growth than non-survivors of the same population. The aim is to address the question of whether growth and development data of non-survivors are reflective of the biological consequences of selective mortality and/or stress. The study compares dental development and skeletal growth collected from radiographic images of contemporary samples of living and deceased individuals from the United States (birth to 20 years) and South Africa (birth to 12 years). Further evaluation of deceased individuals is used to explore differential patterns among manners of death (MOD). Results do not show any significant differences in skeletal growth or dental development between living and deceased individuals. However, in the South African deceased sample the youngest individuals exhibited substantially smaller diaphyseal lengths than the living sample, but by 2 years of age the differences were negligible. In the US sample, neither significant nor substantial differences were found in dental development or diaphyseal length according to MOD and age (>2 years of age), though some long bones in individuals <2 years of age did show significant differences. No significant differences were noted in diaphyseal length according to MOD and age in the SA sample. The current findings refute the idea that contemporary deceased and living individuals would present with differential growth and development patterns through all of ontogeny as well as the assumptions linking short stature, poor environments, and MOD.


A peroxidase biomimetic system based on Fe3O4 nanoparticles in non-enzymatic sensors.
Magnetic nanoparticles have been applied in many areas of nanomedicine. Sensing platforms based on this type of nanoparticles have received attention due to the relative low cost and biocompatibility. Biosensor is the most widely investigated type of analytical device. This type of sensor combines the physicochemical transduction with the incorporation of biological sensing components. Among the biological components, enzymes are the most commonly used as sensitive elements. However, natural enzymes may exhibit serious disadvantages such as lack of stability and loss of catalytic activity after immobilization. The study of enzymatic biomimetic systems are of great interest. This study reports the development of a new sensor composed of Fe3O4@CTAB and poly(sodium 4-styrenesulfonate) (PSS) films assembled via Layer-by-Layer (LbL) technique and used as peroxidase mimetic systems. Magnetic nanoparticles (MNps) were synthesized using thermal decomposition method and further dispersed to aqueous medium by ligand modification reaction using cetyltrimethylammonium bromide (CTAB). The amperometric detection limit of H2O2 was found to be ca. 103 µmol L(-1). By chronoamperometry, the peroxidase biomimetic sensor exhibited a linear response for H2O2 in the range from 100 µmol L(-1) to 1.8 mmol L(-1) (R(2) = 0.994) with sensitivity of 16 nA mol(-1) L. The apparent Michaelis-Menten constant was 5.3 mmol L(-1), comparable with some biosensors based on peroxidase enzyme. Moreover, the sensor presented a reproducibility of ca. 7.7% (n = 4) and their response (response time: 90 s) is not significantly affected in the presence of some interferents including K(+), Na(+), Cl(-), Mg(2+), Ca(2+), and Uric Acid.


Exciton states of dynamic DNA double helices: alternating dCdG sequences.
The present communication deals with the excited states of the alternating DNA oligomer (dCdG)5.(dCdG)5 which correspond to the UV absorption band around 260 nm. Their properties are studied in the frame of the exciton theory, combining molecular dynamics simulations and quantum chemistry data. It is shown that the dipolar coupling undergoes important variations with the site and the helix geometry. In contrast, the energy of the monomer transitions within the double helix is not sensitive to the local environment. It is thus considered to be distributed over Gaussian curves whose maximum and width are derived from the experimental absorption spectra of nucleosides in aqueous solution. The influence of the spectral width on the excited state delocalization and the absorption spectra is much stronger than that of the oligomer plasticity. About half of the excited states are delocalized over at least two bases. Many of them result from the mixing of different monomer states and extend on both strands. The trends found in the simulated spectra, when going from non-interacting monomers to the duplex, are in agreement with experimental observations. Conformational changes enhance the diversity of the states which can be populated upon excitation at a given energy. The states with larger spatial extent are located close to the maximum of the absorption spectrum.


Effects of second-generation antipsychotic medication on smooth pursuit performance in antipsychotic-naive schizophrenia.
Analyses of smooth pursuit eye movement parameters in patients with schizophrenia provide information about the integrity of neural networks mediating motion perception, sensorimotor transformation, and cognitive processes such as prediction. Although pursuit eye tracking deficits have been widely reported in schizophrenia, the integrity of discrete components of pursuit responses and the effect of second-generation antipsychotic medication on them are not well established. To examine different components of smooth pursuit performance in antipsychotic-naive patients with schizophrenia before and after treatment with second-generation antipsychotic medication. Thirty-three antipsychotic-naive patients with schizophrenia performed 3 different smooth pursuit paradigms designed to evaluate specific components of the pursuit response. All of the patients were retested after 6 weeks of treatment with risperidone or olanzapine. Testing was also performed with 39 matched healthy individuals. Thirteen patients and 21 healthy participants were retested after 26 and 52 weeks. Pursuit initiation, maintenance gain (ratio of eye velocity over target velocity), and frequency of catch-up saccades during pursuit maintenance. Prior to treatment, pursuit gain when tracking less predictable ramp targets tended to be reduced, latency of pursuit initiation was speeded, and catch-up saccade frequency was increased during predictive pursuit. After antipsychotic treatment initiation, pursuit gain decreased with ramp targets, indicating treatment-emergent impairments in sensorimotor processing. No changes were observed for predictive pursuit. Exploratory analyses in the subgroup with follow-up to 1 year revealed that these effects continued through long-term follow-up with some partial normalization at 1 year. Deficits were unrelated to drug dosage and clinical ratings. Impaired sensorimotor function was observed after initiation of second-generation antipsychotic medications, which may be explained by their serotonergic antagonism of brainstem sensorimotor systems. Predictive mechanisms supported by frontostriatal-cerebellar circuitry were not affected by treatment initiation and appear able to compensate for treatment-emergent sensorimotor impairments during predictive tracking.


High health care costs in minority groups of older US Medicare beneficiaries with epilepsy.
To examine health care costs in diverse older Medicare beneficiaries with epilepsy. Using 2008-2010 claims data, we conducted a longitudinal cohort study of a random sample of Medicare beneficiaries augmented for minority representation. Epilepsy cases (n = 36 912) had ≥1 International Classification of Diseases, Ninth Edition (ICD-9) 345.x or ≥2 ICD-9 780.3x claims, and ≥1 antiepileptic drug (AED) in 2009; new cases (n = 3706) had no seizure/epilepsy claims nor AEDs in the previous 365 days. Costs were measured by reimbursements for all care received. High cost was defined as follow-up 1-year cost ≥ 75th percentile. Logistic regressions examined association of high cost with race/ethnicity, adjusting for demographic, clinical, economic, and treatment quality factors. In cases with continuous 2-year data, we obtained costs in two 6-month periods before and two after the index event. Cohort was ~62% African Americans (AAs), 11% Hispanics, 5% Asians, and 2% American Indian/Alaska Natives. Mean costs in the follow-up were ~$30 000 (median = $11 547; new cases, mean = $44 642; median = $25 008). About 19% white compared to 27% AA cases had high cost. AA had higher odds of high cost in adjusted analyses (odds ratio [OR] = 1.20, 95% confidence interval [CI] = 1.11-1.29), although this was only marginally significant when adjusting for AED adherence (OR = 1.09, 95% CI = 1.01-1.18, P = 0.03). Factors associated with high cost included ≥1 comorbidity, neurological care, and low AED adherence. Costs were highest at ~$17 000 in the 6 months immediately before and after the index event (>$29 000 for new cases). The financial sequelae of epilepsy among older Americans disproportionally affect minorities. Studies should examine contributors to high costs.


Development of a higher power cooling system for lithium targets.
The accelerator based Boron Neutron Capture Therapy beam at the University of Birmingham is based around a solid thick lithium target cooled by heavy water. Significant upgrades to Birmingham's Dynamitron accelerator are planned prior to commencing a clinical trial. These upgrades will result in an increase in maximum achievable beam current to at least 3 mA. Various upgrades to the target cooling system to cope with this increased power have been investigated. Tests of a phase change coolant known as "binary ice" have been carried out using an induction heater to provide a comparable power input to the Dynamitron beam. The experimental data shows no improvement over chilled water in the submerged jet system, with both systems exhibiting the same heat input to target temperature relation for a given flow rate. The relationship between the cooling circuit pumping rate and the target temperature in the submerged jet system has also been tested.


Phylogeographic insights into an irruptive pest outbreak.
Irruptive forest insect pests cause considerable ecological and economic damage, and their outbreaks have been increasing in frequency and severity. We use a phylogeographic approach to understand the location and progression of an outbreak by the MPB (Dendroctonus ponderosae Hopkins), an irruptive bark beetle that has caused unprecedented damage to lodgepole pine forests in western North America and is poised to expand its range across the boreal forest. We sampled MPB populations across British Columbia and Alberta and used phylogeographic methods to describe lineage diversification, characterize population structure, investigate expansion dynamics, and identify source populations of the outbreak. Using 1181 bp of mitochondrial DNA sequence from 267 individuals, we found high haplotype diversity, low nucleotide diversity, and limited lineage diversification. The overall pattern was consistent with isolation by distance at a continental scale, and with reduced diversity and population structure in the northerly, outbreak regions. Post-Pleistocene expansion was detected, however more recent expansion signals were not detected, potentially due to the size and rapid rate of range expansion. Based on the limited genetic structure, there were likely multiple source populations in southern British Columbia, although the magnitude of the demographic expansion and rate of spread have obscured the signature of these source populations. Our data highlight the need for caution in interpreting phylogeographic results for species with similar demographics.


Robotic versus laparoscopic radical hysterectomy in cervical cancer patients: a matched-case comparative study.
This study aimed to compare initial surgical outcomes and complication rates of patients with early-stage cervical cancer who underwent robotic radical hysterectomy (RRH) and conventional laparoscopic radical hysterectomy (LRH). Patients diagnosed with invasive cervical cancer (International Federation of Gynecology and Obstetrics stage I-IIA) who underwent RRH (n = 23) at Samsung Medical Center from January 2008 to May 2013 were compared with matched patients who underwent LRH (n = 69) during the same period. The 2 surgical groups were matched 3:1 for variables of age, body mass index, International Federation of Gynecology and Obstetrics stage, histological subtype, tumor size, and node positivity. All patient information and surgical and postoperative follow-up data were retrospectively collected. Operating time was significantly longer (317 vs 236 minutes; P < 0.001) in the RRH group compared with the LRH group but mean estimated blood loss was significantly reduced in the RRH group (200 vs 350 mL; P = 0.036). Intraoperative and postoperative complications were not significantly different between the 2 groups (4.3% for RRH vs 1.45% for LRH; P = 0.439). Recurrences were 2 (8.7%) in the RRH and 7 (10.1%) in the LRH group. The overall 3-year recurrence-free survival was 91.3% in RRH group and 89.9% in the LRH group (P = 0.778). Although operating time was longer in the RRH cases because of lesser experience on robotic platform, we showed that surgical outcomes and complication rate of RRH were comparable to those of LRH. In addition, surgical skills for LRH easily and safely translated to RRH in case of experienced laparoscopic surgeon.


Olive oil and leaf extract prevent fluoxetine-induced hepatotoxicity by attenuating oxidative stress, inflammation and apoptosis.
Olive oil and leaf extract have several health benefits; however, their beneficial effect against fluoxetine-induced liver injury has not been investigated. The present study aimed to scrutinize the impact of fluoxetine on the liver of rats and to evaluate the protective effects of olive oil and leaf extract. Rats received fluoxetine orally at dose of 10 mg/kg body weight for 7 consecutive days. The fluoxetine-induced rats were concurrently treated with olive oil or leaf extract. At the end of the experiment, blood and liver samples were collected for analysis. Fluoxetine administration significantly increased circulating ALT, AST, ALP and the pro-inflammatory cytokines TNF-α and IL-1β levels in rats. Histological analysis showed several alterations, such as inflammatory cells infiltration, hepatocyte vacuolation and dilated sinusoids in the liver of fluoxetine-induced rats. Concurrent supplementation of olive oil and olive leaf extract significantly reduced circulating liver function marker enzymes and pro-inflammatory cytokines, and prevented fluoxetine-induced histological alterations. Both olive oil and leaf extract significantly decreased liver lipid peroxidation and nitric oxide, and ameliorated liver glutathione, superoxide dismutase, catalase and glutathione peroxidase. In addition, olive oil and leaf extract prevented fluoxetine-induced apoptosis in the liver of rats as evidenced by decreased expression of Bax and caspase-3, and up-regulated expression of Bcl-2. In conclusion, olive oil and leaf extract protect against fluoxetine-induced liver injury in rats through attenuation of oxidative stress, inflammation and apoptosis.


Mathematical modelling of pressure-driven micropolar biological flow due to metachronal wave propulsion of beating cilia.
In this paper, we present an analytical study of pressure-driven flow of micropolar non-Newtonian physiological fluids through a channel comprising two parallel oscillating walls. The cilia are arranged at equal intervals and protrude normally from both walls of the infinitely long channel. A metachronal wave is generated due to natural beating of cilia and the direction of wave propagation is parallel to the direction of fluid flow. Appropriate expressions are presented for deformation via longitudinal and transverse velocity components induced by the ciliary beating phenomenon with cilia assumed to follow elliptic trajectories. The conservation equations for mass, longitudinal and transverse (linear) momentum and angular momentum are reduced in accordance with the long wavelength and creeping Stokesian flow approximations and then normalized with appropriate transformations. The resulting non-linear moving boundary value problem is solved analytically for constant micro-inertia density, subject to physically realistic boundary conditions. Closed-form expressions are derived for axial velocity, angular velocity, volumetric flow rate and pressure rise. The transport phenomena are shown to be dictated by several non-Newtonian parameters, including micropolar material parameter and Eringen coupling parameter, and also several geometric parameters, viz eccentricity parameter, wave number and cilia length. The influence of these parameters on streamline profiles (with a view to addressing trapping features via bolus formation and evolution), pressure gradient and other characteristics are evaluated graphically. Both axial and angular velocities are observed to be substantially modified with both micropolar rheological parameters and furthermore are significantly altered with increasing volumetric flow rate. Free pumping is also examined. An inverse relationship between pressure rise and flow rate is computed which is similar to that observed in Newtonian fluids. The study is relevant to hemodynamics in narrow capillaries and also bio-inspired micro-fluidic devices.


Muscle damage during minimally invasive total hip arthroplasty: cadaver-based evidence that it is significant.
Minimally invasive total hip arthroplasty has generated substantial interest in both patients and surgeons. The concept that smaller incisions and less extensive surgical dissection should lead to less pain and a quicker recovery is inherently appealing. Advocates of minimally invasive total hip arthroplasty have suggested that some minimally invasive total hip approaches can be done without cutting any muscle or tendon. This contention has been carefully examined through a series of comparative cadaver studies, and the authors have determined that it is not possible to routinely perform minimally invasive total hip arthroplasty without causing some measurable degree of muscle damage. Anatomic studies showed that the two-incision approach with fluoroscopy, the posterior mini-incision approach, and the mini-incision Smith-Petersen approach all were associated with measurable muscle damage. The clinical importance of this muscle damage cannot be answered by these cadaver studies.


HMO enrollment: a study of market forces and regulations.
This paper investigates the influence of market forces and state regulations on enrollment in prepaid group practices (PGPs)--the dominant form of HMO. Using data at the metropolitan-area level, the paper estimates a lagged-adjustment model in two stages. The first stage estimates the determinants of the existence of a PGP of viable size. Using a technique appropriate for censored samples, the second stage estimates the determinants of enrollment given viability. The result of greatest relevance for policymakers is that restrictions on corporate employment of physicians hinder the growth of PGPs.


A model for abnormal activity recognition and alert generation system for elderly care by hidden conditional random fields using R-transform and generalized discriminant analysis features.
The growing population of elderly people living alone increases the need for automatic healthcare monitoring systems for elderly care. Automatic vision sensor-based systems are increasingly used for human activity recognition (HAR) in recent years. This study presents an improved model, tested using actors, of a sensor-based HAR system to recognize daily life activities of elderly people at home and generate an alert in case of abnormal HAR. Datasets consisting of six abnormal activities (falling backward, falling forward, falling rightward, falling leftward, chest pain, and fainting) and four normal activities (walking, rushing, sitting down, and standing up) are generated from different view angles (90°, -90°, 45°, -45°). Feature extraction and dimensions reduction are performed by R-transform followed by generalized discriminant analysis (GDA) methods. R-transform extracts symmetric, scale, and translation-invariant features from the sequences of activities. GDA increases the discrimination between different classes of highly similar activities. Silhouette sequences are quantified by the Linde-Buzo-Gray algorithm and recognized by hidden conditional random fields. Experimental results provide an average recognition rate of 94.2% for abnormal activities and 92.7% for normal activities. The recognition rate for the highly similar activities from different view angles shows the flexibility and efficacy of the proposed abnormal HAR and alert generation system for elderly care.


Transient elastography for liver fibrosis diagnosis.
Liver biopsy is considered the "golden standard" for assessment of hepatic fibrosis. However, the procedure has limitations because of inconvenience and rare but serious complications as bleeding. Furthermore, sampling errors are frequent, and interobserver variability often poses problems. Recently, a modified ultrasound scanner (transient elastography) has been developed to assess fibrosis. The device measures liver elasticity, which correlates well with the degree of fibrosis. Studies have shown that transient elastography is more accurate in diagnosing cirrhosis than minor to moderate fibrosis. Most of the studies have been conducted on patients with chronic hepatitis but a few studies have also covered fibrosis and cirrhosis due to other etiologies, and they also demonstrate the high sensitivity and specificity. Transient elastography for assessment of fibrosis may turn out to be a valuable diagnostic procedure and follow-up of patients with chronic liver diseases.


Purification and characterization of a yeast DNA polymerase alpha complex with associated primase, 5'-->3' exonuclease, and DNA-dependent ATPase activities.
We have purified a multimeric form of yeast DNA polymerase alpha with DNA polymerase, primase, 5'-->3' exonuclease, and single-stranded (ss) DNA-dependent ATPase activities to near-homogeneity. The molecular mass of complex was 650 kDa with subunits ranging in sizes from 30 to 180 kDa. The alpha-subunit of the complex could be detected by DNA polymerase alpha antibody. No cross-reactivity of polypeptides within the complex was observed with antibodies directed against polymerase delta or epsilon. The multimeric polymerase alpha could be selectively inhibited by p-n-butylphenyl-dGTP (I50 of approximately 0.2 microM), p-n-butylanilino-dATP (I50 of 1.3 microM), and aphidicolin (I50 of 2.5 micrograms/mL). The complex synthesized RNA primers on various ssDNA templates and rapidly elongated these primers into nascent DNA fragments in the presence of required deoxynucleotides. It has a strong 5'-->3' exonuclease activity. In addition, the complex hydrolyzed both ATP and dATP in a ssDNA-dependent manner. Thus, the multiprotein complex of DNA polymerase alpha had multiple activities (primase, polymerase, and ATPase) which could act concertedly to synthesize primers and elongate the primers to nascent DNA fragments in the lagging strand of the fork.


Membrane fusion mutants of Semliki Forest virus.
Previous reports have indicated that the entry of Semliki Forest virus (SFV) into cells depends on a membrane fusion reaction catalyzed by the viral spike glycoproteins and triggered by the low pH prevailing in the endosomal compartment. In this study the in vitro pH-dependent fusion of SFV with nuclease-filled liposomes has been used to select for a new class of virus mutants that have a pH-conditional defect. The mutants obtained had a threshold for fusion of pH 5.5 as compared with the wild-type threshold of 6.2, when assayed by polykaryon formation, fusion with liposomes, or fusion at the plasma membrane. They were fully capable of infecting cells under standard infection conditions but were more sensitive to lysosomotropic agents that increase the pH in acidic vacuoles of the endocytic pathway. The mutants were, moreover, able to penetrate and infect baby hamster kidney-21 cells at 20 degrees C, indicating that the endosomes have a pH below 5.5. The results confirm the involvement of pH-triggered fusion in SFV entry, emphasize the central role played by acidic endosomal vacuoles in this reaction, shed further light on the mechanism of SFV inhibition by lysosomotropic weak bases, and demonstrate the usefulness of mutant viruses as biological pH probes of the endocytic pathway.


A Contemporary Medicolegal Analysis of Implanted Devices for Chronic Pain Management.
Analysis of closed malpractice claims allows the study of rare but serious complications and likely results in improved patient safety by raising awareness of such complications. Clinical studies and closed claims analyses have previously reported on the common complications associated with intrathecal drug delivery systems (IDDS) and spinal cord stimulators (SCS). This study provides a contemporary analysis of claims from within the past 10 years. We performed a closed claims analysis for a period January 1, 2009 to December 31, 2013 for cases with pain medicine as the primary service. These cases were identified using the Controlled Risk Insurance Company (CRICO) Comparative Benchmarking System (CBS), a database containing more than 300,000 malpractice claims from more than 400 academic and community institutions, representing approximately 30% of malpractice cases in the United States. The clinical narratives, which included medical files, claims files, depositions, and expert witness testimony, were reviewed by the authors, as were the CRICO coded variables, which included algorithmically determined contributing factors to the patient injury. Intrathecal drug delivery systems represented 17 of the closed claims; spinal cord stimulators represented 11 of the closed claims. The most common chronic pain diagnoses for which a device was implanted included failed back surgery syndrome and spasticity. The average total incurred for pain medicine claims was $166,028. Damaging events included IDDS refill errors (eg, subcutaneous administration of medication, reprogramming errors), intraoperative nerve damage, and postoperative infection (eg, epidural abscess, meningitis). High-severity outcomes included nerve damage (eg, paraplegia) and death. Medium-severity outcomes included drug reactions (eg, respiratory arrest from opioid overdose) and the need for reoperation. For both IDDS and SCS, deficits in technical skill were the most common contributing factor to injury, followed by deficits in clinical judgment, communication, and documentation. Implanted devices used for pain management involve a significant risk of morbidity and mortality. Proper education of providers and patients is essential. Providers must acquire the technical skills required for the implantation and refilling of these devices and the clinical skills required for the identification and management of complications such as intrathecal granuloma. Proper patient selection and clear communication between the provider and the patient about these possible complications are of paramount importance.


Field Measurements Indicate Unexpected, Serious Underestimation of Mussel Heart Rates and Thermal Tolerance by Laboratory Studies.
Attempts to predict the response of species to long-term environmental change are generally based on extrapolations from laboratory experiments that inevitably simplify the complex interacting effects that occur in the field. We recorded heart rates of two genetic lineages of the brown mussel Perna perna over a full tidal cycle in-situ at two different sites in order to evaluate the cardiac responses of the two genetic lineages present on the South African coast to temperature and the immersion/emersion cycle. "Robomussel" temperature loggers were used to monitor thermal conditions at the two sites over one year. Comparison with live animals showed that robomussels provided a good estimate of mussel body temperatures. A significant difference in estimated body temperatures was observed between the sites and the results showed that, under natural conditions, temperatures regularly approach or exceed the thermal limits of P. perna identified in the laboratory. The two P. perna lineages showed similar tidal and diel patterns of heart rate, with higher cardiac activity during daytime immersion and minimal values during daytime emersion. Comparison of the heart rates measured in the field with data previously measured in the laboratory indicates that laboratory results seriously underestimate heart rate activity, by as much as 75%, especially during immersion. Unexpectedly, field estimates of body temperatures indicated an ability to tolerate temperatures considered lethal on the basis of laboratory measurements. This suggests that the interaction of abiotic conditions in the field does not necessarily raise vulnerability to high temperatures.


Simultaneous inference of cancer pathways and tumor progression from cross-sectional mutation data.
Recent cancer sequencing studies provide a wealth of somatic mutation data from a large number of patients. One of the most intriguing and challenging questions arising from this data is to determine whether the temporal order of somatic mutations in a cancer follows any common progression. Since we usually obtain only one sample from a patient, such inferences are commonly made from cross-sectional data from different patients. This analysis is complicated by the extensive variation in the somatic mutations across different patients, variation that is reduced by examining combinations of mutations in various pathways. Thus far, methods to reconstruct tumor progression at the pathway level have restricted attention to known, a priori defined pathways. In this work we show how to simultaneously infer pathways and the temporal order of their mutations from cross-sectional data, leveraging on the exclusivity property of driver mutations within a pathway. We define the pathway linear progression model, and derive a combinatorial formulation for the problem of finding the optimal model from mutation data. We show that with enough samples the optimal solution to this problem uniquely identifies the correct model with high probability even when errors are present in the mutation data. We then formulate the problem as an integer linear program (ILP), which allows the analysis of datasets from recent studies with large numbers of samples. We use our algorithm to analyze somatic mutation data from three cancer studies, including two studies from The Cancer Genome Atlas (TCGA) on large number of samples on colorectal cancer and glioblastoma. The models reconstructed with our method capture most of the current knowledge of the progression of somatic mutations in these cancer types, while also providing new insights on the tumor progression at the pathway level.


Clinical trial design in adult reflux disease: a methodological workshop.
The development of well-tolerated acid suppressant drugs has stimulated substantial growth in the number of trials assessing therapy options for gastro-oesophageal reflux disease (GERD). To develop consensus statements to inform clinical trial design in adult patients with GERD. Draft statements were developed employing a systematic literature review. A modified Delphi process including three rounds of voting was used to reach consensus. Between voting, statements were revised based on feedback from the Working Group and additional literature reviews. The final vote was at a face-to-face meeting that included discussion time. Voting was conducted using a six-point scale. At the last vote, 93% of the final 102 statements achieved consensus (defined a priori as being supported by >or=75% of the votes). The Working Group strongly supported the development of validated patient-reported outcome instruments. Symptom assessments carried out by the investigator were considered unacceptable. There was agreement that exclusion from clinical trials should be minimized to improve generalizability, that prospective evaluation ideally requires electronic timed/dated methods and that endoscopists should be blinded to patient symptom status. Implementation of the consensus statements will improve the quality and comparability of trials, and make them compatible with regulatory requirements.


A stromal address code defined by fibroblasts.
To navigate into and within tissues, leukocytes require guidance cues that enable them to recognize which tissues to enter and which to avoid. Such cues are partly provided at the time of extravasation from blood by an endothelial address code on the luminal surface of the vascular endothelium. Here, we review the evidence that fibroblasts help define an additional stromal address code that directs leukocyte behaviour within tissues. We examine how this stromal code regulates site-specific leukocyte accumulation, differentiation and survival in a variety of physiological stromal niches, and how the aberrant expression of components of this code in the wrong tissue at the wrong time contributes to the persistence of chronic inflammatory diseases.


Changes of Swimmers' Emotional States during the Preparation of National Championship: Do Recovery-Stress States Matter?
This study examined the trajectories of emotional states and their within-person synergies with perceived stress and recovery during a 4-month training period preceding the French swimming championships. A Multilevel Growth Curve Analysis approach was used with 16 high level swimmers. Five waves of assessments of emotional states, perceived stress and recovery were completed. Results indicated that emotional states were characterized by distinct trajectories during the training period preceding a major competition. Specifically, significant positive linear effects of time (i.e., linear increase over time) and negative quadratic effects of squared time (i.e., inverted U shape over time) on anxiety, dejection and anger were observed, whereas the opposite pattern of results was found for happiness and excitement. Moreover, level 2 perceived stress and recovery (i.e., inter-individual predictors) were significantly associated with athletes' unpleasant and pleasant emotional states respectively. At level 1, perceived recovery (i.e., intra-individual predictor) was positively associated with happiness and excitement and negatively related to anxiety, dejection and anger. Finally, within-person interactions of general stress and recovery with time and squared time reached significance for excitement, whereas within-person interactions of specific and total stress with time and squared time reached significance for anxiety. Overall, this study provided insights into the central role played by perceived stress and recovery on the emotional states experienced by high level swimmers. Operational strategies were suggested in order to optimize the stress-recovery balance and in turn the athletes' emotional states during a complete training program.


The 'azirine/oxazolone method' in peptaibol synthesis: preparation of a derivative of trichotoxin A-50 (G).
The synthesis of a mixture of epimeric derivatives of the peptaibol trichotoxin A-50 (G) is described. The 'azirine/oxazolone method' has been used as a superior method for the introduction of the Aib as well as the Iva units into the peptide chain. In this protocol, 2,2-disubstituted 2H-azirin-3-amines are the synthons for 2,2-disubstituted glycines, which undergo coupling with N-protected amino or peptide acids in high yield, and without any need of coupling reagents. The problem of the instability of the amide function of the Gln side chain under the conditions of the acid-catalyzed hydrolysis of Z-Gln-(Aib)(n)-N(Me)Ph was solved by using an appropriate protecting group for the amide function of the Gln side chain, e.g., the triphenylmethyl (trityl; Tr) group. The structures of two intermediate peptides, i.e., the segments comprising residues 1-5 and 10-13, resp., were established by X-ray crystallography.


Natural Scales in Geographical Patterns.
Human mobility is known to be distributed across several orders of magnitude of physical distances, which makes it generally difficult to endogenously find or define typical and meaningful scales. Relevant analyses, from movements to geographical partitions, seem to be relative to some ad-hoc scale, or no scale at all. Relying on geotagged data collected from photo-sharing social media, we apply community detection to movement networks constrained by increasing percentiles of the distance distribution. Using a simple parameter-free discontinuity detection algorithm, we discover clear phase transitions in the community partition space. The detection of these phases constitutes the first objective method of characterising endogenous, natural scales of human movement. Our study covers nine regions, ranging from cities to countries of various sizes and a transnational area. For all regions, the number of natural scales is remarkably low (2 or 3). Further, our results hint at scale-related behaviours rather than scale-related users. The partitions of the natural scales allow us to draw discrete multi-scale geographical boundaries, potentially capable of providing key insights in fields such as epidemiology or cultural contagion where the introduction of spatial boundaries is pivotal.


Detergent-free incorporation of a seven-transmembrane receptor protein into nanosized bilayer Lipodisq particles for functional and biophysical studies.
SMA-Lipodisq nanoparticles, with one bacteriorhodopsin (bR) per 12 nm particle on average (protein/lipid molar ratio, 1:172), were prepared without the use of detergents. Using pulsed and continuous wave nitroxide spin label electron paramagnetic resonance, the structural and dynamic integrity of bR was retained when compared with data for bR obtained in the native membrane and in detergents and then with crystal data. This indicates the potential of Lipodisq nanoparticles as a useful membrane mimetic.


Idiopathic pulmonary fibrosis and occupational risk factors.
Idiopathic pulmonary fibrosis (IPF) is a rare lung disease of unknown origin that rapidly leads to death. However, the rate of disease progression varies from one individual to another and is still difficult to predict. The prognosis of IPF is poor, with a median survival of three to five years after diagnosis, without curative therapies other than lung transplantation. The factors leading to disease onset and progression are not yet completely known. The current disease paradigm is that sustained alveolar epithelial micro-injury caused by environmental triggers (e.g., cigarette smoke, microaspiration of gastric content, particulate dust, viral infections or lung microbial composition) leads to alveolar damage resulting in fibrosis in genetically susceptible individuals. Numerous epidemiological studies and case reports have shown that occupational factors contribute to the risk of developing IPF. In this perspective, we briefly review the current understanding of the pathophysiology of IPF and the importance of occupational factors in the pathogenesis and prognosis of the disease. Prompt identification and elimination of occult exposure may represent a novel treatment approach in patients with IPF.


Exploring the link between racial discrimination and substance use: what mediates? What buffers?
The relation between perceived racial discrimination and substance use was examined in 2 studies that were based on the prototype-willingness model (Gibbons, Gerrard, & Lane, 2003). Study 1, using structural equation modeling, revealed prospective relations between discrimination and use 5 years later in a panel of African American adolescents (M age 10.5 years at Time 1 [T1]) and their parents. For both groups, the relation was mediated by anger and/or hostility. For the adolescents, it was also mediated by behavioral willingness, and it was moderated by supportive parenting. Study 2 was a lab experiment in which a subset of the Study 1 adolescents (M age = 18.5 years) was asked to imagine a discriminatory experience, and then their affect and drug willingness were assessed. As in the survey study, discrimination was associated with more drug willingness, and that relation was again mediated by anger and moderated by supportive parenting. Implications of the results for research and interventions involving reactions to racial discrimination are discussed.


Voltage-gated potassium channel autoimmunity mimicking creutzfeldt-jakob disease.
Rapidly progressive dementia has a variety of causes, including Creutzfeldt-Jakob disease (CJD) and neuronal voltage-gated potassium channel (VGKC) autoantibody-associated encephalopathy. To describe patients thought initially to have CJD but found subsequently to have immunotherapy-responsive VGKC autoimmunity. Observational, prospective case series. Department of Neurology, Mayo Clinic, and the Memory and Aging Center, University of California, San Francisco. Patients A clinical serologic cohort of 15 patients referred for paraneoplastic autoantibody evaluation. Seven patients were evaluated clinically by at least one of us. Clinical information for the remaining patients was obtained by physician interview or medical record review. Clinical features, magnetic resonance imaging abnormalities, electroencephalographic patterns, cerebrospinal fluid analyses, and responses to immunomodulatory therapy. All the patients presented subacutely with neurologic manifestations, including rapidly progressive dementia, myoclonus, extrapyramidal dysfunction, visual hallucinations, psychiatric disturbance, and seizures; most (60%) satisfied World Health Organization diagnostic criteria for CJD. Magnetic resonance imaging abnormalities included cerebral cortical diffusion-weighted imaging hyperintensities. Electroencephalographic abnormalities included diffuse slowing, frontal intermittent rhythmic delta activity, and focal epileptogenic activity but not periodic sharp wave complexes. Cerebrospinal fluid 14-3-3 protein or neuron-specific enolase levels were elevated in 5 of 8 patients. Hyponatremia was common (60%). Neoplasia was confirmed histologically in 5 patients (33%) and was suspected in another 5. Most patients' conditions (92%) improved after immunomodulatory therapy. Clinical, radiologic, electrophysiologic, and laboratory findings in VGKC autoantibody-associated encephalopathy may be confused with those of CJD. Serologic evaluation for markers of neurologic autoimmunity, including VGKC autoantibodies, may be warranted in suspected CJD cases.


Osmotic blood-brain barrier disruption in the posterior fossa of the dog.
Transient reversible osmotic blood-brain barrier disruption was produced in the posterior fossa of 33 dogs. A percutaneous catheter technique was used for the infusion of hypertonic mannitol into the vertebral artery. Neither the catheter technique nor the osmotic barrier modification resulted in interference with brain-stem function in most animals. The degree of barrier modification achieved by osmotic disruption in the posterior fossa is similar to that previously described for barrier modification of the supratentorial parenchyma. Methotrexate delivered to the brain via the vertebral artery resulted in a drug concentration of 100 to 300 ng/gm brain tissue. When the same amount of drug was given following osmotic blood-brain barrier disruption, brain tissue contained 1100 to 5000 ng of methotrexate/gm of brain tissue. Finally, the adequacy of the blood-brain barrier modification in the posterior fossa was shown to be quantifiable by the amount of enhancement on computerized tomographic scans.


Changing Faces: Dynamic Emotional Face Processing in Autism Spectrum Disorder Across Childhood and Adulthood.
Autism spectrum disorder (ASD) is classically associated with poor emotional face processing. Few studies, however, have used more ecological dynamic stimuli. We contrasted functional magnetic resonance imaging measures of dynamic emotional face processing in ASD and typically developing (TD) cohorts across a wide age range to determine if the processing and age-related trajectories differed between participants with and without ASD. Functional magnetic resonance imaging data collected from 200 participants (5-42 years old; 107 in ASD cohort, 93 in TD cohort) during the presentation of dynamic emotional faces (neutral-to-happy, neutral-to-angry) and dynamic flowers (closed-to-open) were analyzed. Group differences and group-by-age interactions in the faces versus flowers and between emotion contrasts were investigated. Differences in activation between dynamic faces and flowers in occipital regions, including the fusiform gyri, were reduced in the ASD group. Contrasting the two emotions, ASD compared with TD participants showed increased engagement of the precentral, postcentral, and superior temporal gyri to happy faces and increased activation to angry faces occipitally. Emotion processing regions, such as insula, temporal pole, and frontal regions, showed increased recruitment with age to happy faces compared with both angry faces and flowers in the TD group, but decreased recruitment with age in the ASD group. Using dynamic stimuli, we demonstrated that participants with ASD processed faces similarly to nonface stimuli, and age-related atypicalities were more pronounced to happy faces in participants with ASD. We demonstrated emotion-specific atypicalities in a large group of participants with ASD that underscore persistent difficulties from childhood into mid-adulthood.


Novel Pactamycin Analogs Induce p53 Dependent Cell-Cycle Arrest at S-Phase in Human Head and Neck Squamous Cell Carcinoma (HNSCC) Cells.
Pactamycin, although putatively touted as a potent antitumor agent, has never been used as an anticancer drug due to its high cytotoxicity. In this study, we characterized the effects of two novel biosynthetically engineered analogs of pactamycin, de-6MSA-7-demethyl-7-deoxypactamycin (TM-025) and 7-demethyl-7-deoxypactamycin (TM-026), in head and neck squamous cell carcinoma (HNSCC) cell lines SCC25 and SCC104. Both TM-025 and TM-026 exert growth inhibitory effects on HNSCC cells by inhibiting cell proliferation. Interestingly, unlike their parent compound pactamycin, the analogs do not inhibit synthesis of nascent protein in a cell-based assay. Furthermore, they do not induce apoptosis or autophagy in a dose- or a time-dependent manner, but induce mild senescence in the tested cell lines. Cell cycle analysis demonstrated that both analogs significantly induce cell cycle arrest of the HNSCC cells at S-phase resulting in reduced accumulation of G2/M-phase cells. The pactamycin analogs induce expression of cell cycle regulatory proteins including master regulator p53, its downstream target p21Cip1/WAF1, p27kip21, p19, cyclin E, total and phospho Cdc2 (Tyr15) and Cdc25C. Besides, the analogs mildly reduce cyclin D1 expression without affecting expression of cyclin B, Cdk2 and Cdk4. Specific inhibition of p53 by pifithrin-α reduces the percentage of cells accumulated in S-phase, suggesting contribution of p53 to S-phase increase. Altogether, our results demonstrate that Pactamycin analogs TM-025 and TM-026 induce senescence and inhibit proliferation of HNSCC cells via accumulation in S-phase through possible contribution of p53. The two PCT analogs can be widely used as research tools for cell cycle inhibition studies in proliferating cancer cells with specific mechanisms of action.


Two types of replication proteins increase the rate at which T4 DNA polymerase traverses the helical regions in a single-stranded DNA template.
We have recently developed an in vitro DNA synthesis system in which a synthetic heptaribonucleotide pairs with a unique site on a single-stranded fd DNA molecule and thereby primes the growth of new DNA strands from this single point (Huang, C.-C., and Hearst, J. E. (1980) Anal. Biochem. 103, 127-139). In this report, we use this system to investigate the mechanism by which various bacteriophage T4 DNA replication proteins stimulate the T4 DNA polymerase. We find that with the "polymerase accessory proteins" present (the T4 gene 44/62 and 45 proteins), the DNA polymerase proceeds rather rapidly through the occasional hairpin helices which otherwise interrupt the progress of this enzyme along single-stranded DNA templates. By using a potent inhibitor of the 44/62 ATPase, ATP gamma S (adenosine 5'-O-(3-thiotriphosphate)), we have obtained data which suggest that ATP hydrolysis is required for the formation of a polymerase accessory protein-DNA template complex, and that this complex then persists, serving as a sliding clamp which greatly increases the strength of binding between a T4 DNA polymerase molecule and its 3'OH primer template end. The progress of the T4 DNA polymerase though hairpin helices in the DNA template is also stimulated by addition of the T4 helix-destabilizing protein (gene 32 protein). The effect of the 44/62 and 45 proteins is independent of the effect of the 32 protein in this assay, and the rate of polymerase travel over the strongest hairpin helices is increased more than 40-fold in the presence of these four additional proteins.


Financial inclusion and intimate partner violence: What does the evidence suggest?
Financial inclusion is an area of growing global interest in women's empowerment policy and programming. While increased economic autonomy may be expected to reduce the prevalence of intimate partner violence, the mechanisms and contexts through which this relationship manifests are not well understood. This analysis aims to assess the relationship between women's financial inclusion and recent intimate partner violence using nationally-representative data from 112 countries worldwide. Levels of both financial inclusion and recent intimate partner violence varied substantially across countries (ranging from 2-100%, and 1-46%, respectively), and across regions. In multivariate global analyses, increased levels of women's financial inclusion were associated with lower levels of recent intimate partner violence after accounting for asset-based enablers of economic autonomy and gender norms; this relationship was lost upon the inclusion of measures of national context (i.e., development and fragility). These results underscore that the relationship between financial inclusion and recent intimate partner violence is complex, follows many pathways, and is affected by context. In low and middle income countries, asset-based enablers of economic autonomy, gender norms and national context explained much of the relationship between financial inclusion and recent intimate partner violence. In those low and middle income countries with high levels of controlling behavior by male spouses, financial inclusion was associated with higher levels of recent intimate partner violence. These findings further suggest that initiatives that aim to prevent intimate partner violence by way of increased economic autonomy may be ineffective in the absence of broader social change and support, and indeed, as seen in countries with higher levels of men's controlling behavior, backlash may increase the risk of violence. Efforts to improve women's financial inclusion need to recognize that its relationship with intimate partner violence is complex, and that it requires an enabling environment supportive of women's rights and autonomy.


Lymphocyte-mediated cytotoxicity in vitro. Effect of enhancing antisera.
The ability of antisera to suppress immune responses either in vivo or in vitro is well known. A variety of lymphocyte-target cell systems have been employed to demonstrate inhibition of cell-mediated immunity by antisera in vitro, and skin, tumor, and kidney graft survival have been prolonged by passively administered antiserum in vivo. An in vitro lymphocyte-tumor cell assay system was developed for the purpose of studying the effects of enhancing antisera (in vivo) on lymphocyte-mediated cytotoxicity in vitro. The characteristics of this system with respect to route of immunization, time of harvest of immune cells, lymphocyte:tumor cell ratio, and effect of nonimmune or nonspecifically immune lymphoid cells are presented. Sera capable of enhancement in vivo were tested in this system and shown to inhibit cell-mediated immunity in vitro. Further, in both instances the immunosuppressive effect is mediated by antigen-antibody complexes and not by free antibody alone. Experiments were also carried out to determine the site of action of these suppressive antigen-antibody complexes. Presensitized lymphocytes were exposed to antigen-antibody complexes, washed, and then allowed to interact with fresh tumor cells (not antibody treated). Lymphocytes treated in this manner are incapable of exhibiting cell-mediated immunity in vitro. This evidence supports the concept that the antigen-antibody complexes have a direct immunosuppressive effect on the lymphocyte.


Design and validity of a clinic-based case-control study on the molecular epidemiology of lymphoma.
We present the design features and implementation of a clinic-based case-control study on the molecular epidemiology of lymphoma conducted at the Mayo Clinic (Rochester, Minnesota, USA), and then assess the internal and external validity of the study. Cases were newly diagnosed lymphoma patients from Minnesota, Iowa and Wisconsin seen at Mayo and controls were patients from the same region without lymphoma who had a pre-scheduled general medical examination, frequency matched on age, sex and residence. Overall response rates were 67% for cases and 70% for controls; response rates were lower for cases and controls over age 70 years, cases with more aggressive disease, and controls from the local area, although absolute differences were modest. Cases and controls were well-balanced on age, sex, and residence characteristics. Demographic and disease characteristics of NHL cases were similar to population-based cancer registry data. Control distributions were similar to population-based data on lifestyle factors and minor allele frequencies of over 500 SNPs, although smoking rates were slightly lower. Associations with NHL in the Mayo study for smoking, alcohol use, family history of lymphoma, autoimmune disease, asthma, eczema, body mass index, and single nucleotide polymorphisms in TNF (rs1800629), LTA (rs909253), and IL10 (rs1800896) were at a magnitude consistent with estimates from pooled studies in InterLymph, with history of any allergy the only directly discordant result in the Mayo study. These data suggest that this study should have strong internal and external validity. This framework may be useful to others who are designing a similar study.


Personality Trait Change in Adulthood.
Recent longitudinal and cross-sectional aging research has shown that personality traits continue to change in adulthood. In this article, we review the evidence for mean-level change in personality traits, as well as for individual differences in change across the life span. In terms of mean-level change, people show increased selfconfidence, warmth, self-control, and emotional stability with age. These changes predominate in young adulthood (age 20-40). Moreover, mean-level change in personality traits occurs in middle and old age, showing that personality traits can change at any age. In terms of individual differences in personality change, people demonstrate unique patterns of development at all stages of the life course, and these patterns appear to be the result of specific life experiences that pertain to a person's stage of life.


The Microbiome and Preterm Birth: A Change in Paradigm with Profound Implications for Pathophysiologic Concepts and Novel Therapeutic Strategies.
Preterm birth poses a global challenge with a continuously increasing disease burden during the last decades. Advances in understanding the etiopathogenesis did not lead to a reduction of prematurely born infants so far. A balanced development of the host microbiome in early life is key for the maturation of the immune system and many other physiological functions. With the tremendous progress in new diagnostic possibilities, the contribution of microbiota changes to preterm birth and the acute and long-term sequelae of prematurity have come into the research focus. This review summarizes the latest advances in the understanding of microbiomes in the amniotic cavity and the female lower genital tract and how changes in microbiota structures contribute to preterm delivery. The exhibition of these highly vulnerable infants to the hostile environment in the neonatal intensive care unit necessarily entails the rapid colonization with a nonbalanced microbiome in a situation where the organism is still very prone and at an early stage of development. The global research efforts to decipher pathologic changes will pave the way to new pre- and postnatal therapeutic concepts.


Disposition and tissue distribution of angiopeptin in the rat.
The disposition and tissue distribution of angiopeptin, a long-acting octapeptide analogue of somatostatin, were studied in rats following single iv and sc administration of the drug. Similar plasma levels and excretion values of angiopeptin were observed by using radioimmunoassay and radiolabeling techniques. Angiopeptin was absorbed fairly rapidly, with a mean peak plasma level of 25 +/- 4.1 ng/ml at 10-15 min after administration. The kinetics of angiopeptin following sc administration closely resembled those following iv administration due to rapid absorption. The pharmacokinetics of angiopeptin can be described by a two-compartment model. The plasma half-life of the drug ranged from 2.6-2.9 hr when administered sc and 1.98-2.5 hr when given iv. Distribution of angiopeptin was rapid, with the highest concentration appearing in the liver. Half-lives in the liver and bile were short. Most of the drug was excreted in the feces via the bile, while approximately 10% was excreted in the urine. Angiopeptin was also found to be secreted in the saliva. TLC and HPLC of blood, urine, feces, and bile samples did not reveal the presence of any metabolites. In conclusion, the in vivo fate of angiopeptin is characterized by little or no hepatic metabolism and rapid biliary excretion.


Utility of Porcine-Derived Xenograft as an Adjunct to Split-Thickness Skin Grafting in Lower-Extremity Wounds.
Porcine-derived xenograft biological dressings (PXBDs) are occasionally used to prepare chronic wound beds for definitive closure before split-thickness skin grafts (STSGs). We sought to determine whether PXBD influences rate of STSG take in lower-extremity wounds. Lower-extremity wounds treated with STSGs were retrospectively reviewed. Patients were included in one of two groups: wound bed preparation with PXBD before STSG or no preparation. Patients were excluded if they received wound bed preparation via another method. Patient demographics, comorbidities, wound history, wound bed preparation, and 30- and 60-day outcomes were collected. There was no difference in healing outcomes between the PXBD (n = 27) and no preparation (n = 39) groups. At 30- and 60-day follow-up, percentage of STSG take was not significantly different between groups (77.9% versus 79.0%, P30 = .818; 82.2% versus 80.9%, P60 = .422). Mean wound sizes at these follow-up periods were not different (4.4 cm2 versus 5.1 cm2, P30 = .902; 1.2 cm2 versus 1.1 cm2, P60 = .689). The PXBD group had a higher mean ± SD hemoglobin A1c level (8.3 ± 3.5 versus 6.9 ± 1.6; P = .074) and age (64.9 ± 12.8 years versus 56.3 ± 11.9 years; P = .007) versus the no preparation group. Application of PXBDs for wound bed preparation had no effect on wound healing compared with no wound bed preparation. The two groups varied only by mean age and hemoglobin A1c level. The PXBD may be beneficial, but these results call for randomized controlled trials to determine the true impact of PXBDs on wound healing. In addition, PXBDs may have utility outside of clinically oriented outcomes, and future work should address patient-reported outcomes and pain scores with this adjunct.


The degradation of curcuminoids in a human faecal fermentation model.
In the present study, the colonic metabolism of three curcuminoids (80.1% curcumin, 15.6%, demethoxycurcumin (DMC) and 2.6% bis-demethoxycurcumin (Bis-DMC)) was evaluated using an in vitro model containing human faecal starters. The breakdown products formed were identified and characterized using different analytical platforms. Following in vitro incubation, the relative amounts of degraded curcuminoids and the produced metabolites were analyzed using a UHPLC coupled with a linear ion trap mass spectrometer, with the addition of hybrid ion trap-Orbitrap Mass Spectrometer when required. Up to ∼24% of curcumin, ∼61% of demethoxycurcumin (DMC) and ∼87% of bis-demethoxycurcumin (Bis-DMC) were degraded by the human faecal microbiota after 24 h of fermentation in vitro. Three main metabolites, namely tetrahydrocurcumin (THC), dihydroferulic acid (DFA) and a metabolite with an accurate mass of 181.08734, which was tentatively identified as 1-(4-Hydroxy-3-methoxyphenyl)-2-propanol were detected in the fermentation cultures containing the curcuminoids. The data presented here provide insights into curcuminoid colonic metabolism, showing that bacterial breakdown products should be considered in further studies on both bioavailability bioactivity of curcumin.


Multimodal Interventions for Caregivers of People With Stroke (January 1, 1999- December 31, 2019).
Systematic Review Briefs provide a summary of the findings from systematic reviews developed in conjunction with the American Occupational Therapy Association's Evidence-Based Practice Program. Each Systematic Review Brief summarizes the evidence on a theme related to a systematic review topic. This Systematic Review Brief presents findings from the systematic review on stroke and multimodal interventions for caregivers of people with stroke.


Fast-track hip and knee arthroplasty: clinical and organizational aspects.
Fast-track hip and knee arthroplasty aims at giving the patients the best available treatment at all times, being a dynamic entity. Fast-track combines evidence-based, clinical features with organizational optimization including a revision of traditions resulting in a streamlined pathway from admission till discharge – and beyond. The goal is to reduce morbidity, mortality and functional convalescence with an earlier achievement of functional milestones including functional discharge criteria with subsequent reduced length of stay and high patient satisfaction. Outcomes are traditionally measured as length of stay; safety aspects in the form of morbidity/mortality; patient satisfaction; and – as a secondary parameter – economic savings. Optimization of the clinical aspects include focusing on analgesia; DVT-prophylaxis; mobilization; care principles including functional discharge criteria; patient-characteristics to predict outcome; and traditions which may be barriers in optimizing outcomes. Patients should be informed and motivated to be active participants and their expectations should be modulated in order to improve satisfaction. Also, organizational aspects need to be analyzed and optimized. New logistical approaches should be implemented; the ward ideally (re)structured to only admit arthroplasties; the staff educated to have a uniform approach; extensive preoperative information given including discharge criteria and intended length of stay. This thesis includes 9 papers on clinical and organizational aspects of fast-track hip and knee arthroplasty (I–IX). A detailed description of the fast-track set-up and its components is provided. Major results include identification of patient characteristics to predict length of stay and satisfaction with different aspects of the hospital stay (I); how to optimize analgesia by using a compression bandage in total knee arthroplasty (II); the clinical and organizational set-up facilitating or acting as barriers for early discharge (III); safety aspects following fast-track in the form of few readmissions in general (IV) and few thromboembolic complications in particular (V); feasibility studies showing excellent outcomes following fast-track bilateral simultaneous total knee arthroplasty (VI) and non-septic revision knee arthroplasty (VII); how acute pain relief in total hip arthroplasty is not enhanced by the use of local infiltration analgesia when multi-modal opioid-sparing analgesia is given (VIII); and a detailed description of which clinical and organizational factors detain patients in hospital following fast-track hip and knee arthroplasty (IX). Economic savings following fast-track hip and knee arthroplasty is also documented in studies, reviews, metaanalyses and Cochrane reviews – including the present fast-track (ANORAK). In conclusion, the published results (I–IX) provide substantial, important new knowledge on clinical and organizational aspects of fast-track hip and knee arthroplasty – with concomitant documented high degrees of safety (morbidity/mortality) and patient satisfaction. Future research strategies are multiple and include both research strategies as efforts to implement the fast-track methodology on a wider basis. Research areas include improvements in pain treatment, blood saving strategies, fluid plans, reduction of complications, avoidance of tourniquet and concomitant blood loss, improved early functional recovery and muscle strengthening. Also, improvements in information and motivation of the patients, preoperative identification of patients needing special attention and detailed economic studies of fast- track are warranted.


Bounding the effect of noise in multiobjective learning classifier systems.
This paper analyzes the impact of using noisy data sets in Pittsburgh-style learning classifier systems. This study was done using a particular kind of learning classifier system based on multiobjective selection. Our goal was to characterize the behavior of this kind of algorithms when dealing with noisy domains. For this reason, we developed a theoretical model for predicting the minimal achievable error in noisy domains. Combining this theoretical model for crisp learners with graphical representations of the evolved hypotheses through multiobjective techniques, we are able to bound the behavior of a learning classifier system. This kind of modeling lets us identify relevant characteristics of the evolved hypotheses, such as overfitting conditions that lead to hypotheses that poorly generalize the concept to be learned.


All-Optical Chirality-Sensitive Sorting via Reversible Lateral Forces in Interference Fields.
Separating substances by their chirality faces great challenges as well as opportunities in chemistry and biology. In this study, we propose an all-optical solution for passive sorting of chiral objects using chirality-dependent lateral optical forces induced by judiciously interfered fields. First, we investigate the optical forces when the chiral objects are situated in the interference field formed by two plane waves with arbitrary polarization states. When the plane waves are either linearly or circularly polarized, nonzero lateral forces are found at the particle's trapping positions, making such sideways motions observable. Although the lateral forces have different magnitudes on particles with different chirality, their directions are the same for opposite handedness particles, rendering it difficult to separate the chiral particles. We further solve the sorting problem by investigating more complicated polarization states. Finally, we achieve the chiral-selective separation by illuminating only one beam toward the chiral substance situated at an interface between two media, taking advantage of the native interference between the incident and reflective beams at the interface. Our study provides a robust and insightful approach to sort chiral substances and biomolecules with plausible optical setups.


Quantitative trait loci for resistance to Haemonchus contortus artificial challenge in Red Maasai and Dorper sheep of East Africa.
A genome-wide scan was performed to detect quantitative trait loci (QTL) for resistance to the gastrointestinal nematode Haemonchus contortus in a double backcross population of Red Maasai and Dorper sheep. The mapping population comprised six sire families, with 1026 lambs in total. The lambs were artificially challenged with H. contortus at about 6.5 months of age, and nine phenotypes were measured: fecal egg count, packed cell volume decline, two weight traits and five worm traits. A subset of the population (342 lambs) was selectively genotyped for 172 microsatellite loci covering 25 of the 26 autosomes. QTL mapping was performed for models which assumed that the QTL alleles were either fixed or segregating within each breed, combined with models with only an additive QTL effect fitted or both additive and dominance QTL effects fitted. Overall, QTL significant at the 1% chromosome-wide level were identified for 22 combinations of trait and chromosome. Of particular interest are a region of chromosome 26 with putative QTL for all nine traits and a region of chromosome 2 with putative QTL for three traits. Favorable QTL alleles for disease resistance originated in both the Red Maasai and Dorper breeds, were not always fixed within breed and had significant dominance effects in some cases. We anticipate that this study, in combination with follow-up work and other relevant studies, will help elucidate the biology of disease resistance.


Estradiol-Mediated Axogenesis of Hypothalamic Neurons Requires ERK1/2 and Ryanodine Receptors-Dependent Intracellular Ca2+ Rise in Male Rats.
17β-estradiol (E2) induces axonal growth through extracellular signal-regulated kinase 1 and 2 (ERK1/2)-MAPK cascade in hypothalamic neurons of male rat embryos in vitro, but the mechanism that initiates these events is poorly understood. This study reports the intracellular Ca2+ increase that participates in the activation of ERK1/2 and axogenesis induced by E2. Hypothalamic neuron cultures were established from 16-day-old male rat embryos and fed with astroglia-conditioned media for 48 h. E2-induced ERK phosphorylation was completely abolished by a ryanodine receptor (RyR) inhibitor (ryanodine) and partially attenuated by an L-type voltage-gated Ca2+ channel (L-VGCC) blocker (nifedipine), an inositol-1,4,5-trisphosphate receptor (IP3R) inhibitor (2-APB), and a phospholipase C (PLC) inhibitor (U-73122). We also conducted Ca2+ imaging recording using primary cultured neurons. The results show that E2 rapidly induces an increase in cytosolic Ca2+, which often occurs in repetitive Ca2+ oscillations. This response was not observed in the absence of extracellular Ca2+ or with inhibitory ryanodine and was markedly reduced by nifedipine. E2-induced axonal growth was completely inhibited by ryanodine. In summary, the results suggest that Ca2+ mobilization from extracellular space as well as from the endoplasmic reticulum is necessary for E2-induced ERK1/2 activation and axogenesis. Understanding the mechanisms of brain estrogenic actions might contribute to develop novel estrogen-based therapies for neurodegenerative diseases.


An HDAC1-binding domain within FATS bridges p21 turnover to radiation-induced tumorigenesis.
There is a gap between the initial formation of cells carrying radiation-induced genetic damage and their contribution to cancer development. Herein, we reveal a previously uncharacterized gene FATS through a genome-wide approach and demonstrate its essential role in regulating the abundance of p21 in surveillance of genome integrity. A large exon coding the NH2-terminal domain of FATS, deleted in spontaneous mouse lymphomas, is much more frequently deleted in radiation-induced mouse lymphomas. Its human counterpart is a fragile site gene at a previously identified loss of heterozygosity site. FATS is essential for maintaining steady-state level of p21 protein and sustaining DNA damage checkpoint. Furthermore, the NH2-terminal FATS physically interacts with histone deacetylase 1 (HDAC1) to enhance the acetylation of endogenous p21, leading to the stabilization of p21. Our results reveal a molecular linkage between p21 abundance and radiation-induced carcinogenesis.


Nuclear morphometry and prognosis in favorable histology Wilms' tumor: A prospective reevaluation.
This study was designed to evaluate the ability of a previously published nuclear morphometry discriminant function to predict disease-free survival in patients with Wilms' tumor. We identified 218 patients with stage I-IV Wilms' tumor of favorable histology who were entered onto the National Wilms' Tumor Study (NWTS) between January 1, 1990 and April 15, 1994. The nuclear morphometry score was calculated for each patient as follows: MV(f) = (0.02 x AGE) + (1.17 x SNRF) + (90.6 x LEFD) - 94, with AGE denoting age at diagnosis in months, SNRF the skewness of the nuclear roundness factor, and LEFD the lowest value of nuclear ellipticity as measured by the feret diameter method. Relative risks of relapse were estimated for the total score and for each of its components. Sensitivity and specificity were determined for the criterion of "MV(f) is greater than -0.35" as a predictor of relapse. By contrast with previously published results, neither the SNRF nor the LEFD made any contribution to the prediction of disease-free survival. Sensitivity and specificity of the criterion of "MV(f) is greater than -0.35" were 71% and 56%, respectively. Re-evaluation of a published nuclear morphometry score showed that it did not predict disease-free survival in patients with Wilms' tumor. The earlier study very likely overestimated the predictive power of nuclear morphometry by using the same data set both to develop the score and to evaluate its properties. Because of the huge number of combinations of nuclear morphometry measurements that may enter into the multivariate discriminant function, use of appropriate statistical methods is essential to estimate accurately the sensitivity and specificity.


Interleukin-6 induces Alzheimer-type phosphorylation of tau protein by deregulating the cdk5/p35 pathway.
Inflammation is a process that has been actively related with the onset of several neurodegenerative disorders including Alzheimer disease (AD). However, the precise implications of inflammatory response for neurodegeneration have not been elucidated. A current hypothesis considers that extracellular insults to neurons could trigger the production of inflammatory cytokines by astrocytes and microglia. These cytokines, namely, interleukin (IL)-1beta, TNFalpha, and IL-6, could affect the normal behavior of neuronal cells. In the present study, we describe the effect of the administration at physiologic doses of one of these cytokines, IL-6, to hippocampal neurons, on the protein kinase pathways as well as on the tau phosphorylation patterns. IL-6-treated neurons exhibited an increase in the amount of anomalously hyperphosphorylated tau protein in epitopes dependent on proline-directed protein kinases (PDPKs). On the basis of our data, the observed increase of tau epitopes of Alzheimer type is explained by an increase of intraneuronal levels of p35 activator and in the activity of the protein kinase cdk5 in response to this cytokine. Further confirmation of cdk5 involvement in this process was based on the findings that inhibition of the kinase activity with butyrolactone-I prevents the appearance of tau of Alzheimer type in IL-6-treated neurons. Additional studies suggest that an increase of cdk5 activity could be mediated by a known signaling cascade described for IL-6 function, namely, the MAPK-p38 signaling pathway. Stimulation of the IL-6 pathway appears to increase the tau epitopes of Alzheimer type, as demonstrated in studies with specific inhibitors. These results support the findings of a pathologic role for IL-6 in the neuroinflammatory response as related with the pathogenesis of neuronal degeneration.


Efficiency of peptide nucleic acid-directed PCR clamping and its application in the investigation of natural diets of the Japanese eel leptocephali.
Polymerase chain reaction (PCR)-clamping using blocking primer and DNA-analogs, such as peptide nucleotide acid (PNA), may be used to selectively amplify target DNA for molecular diet analysis. We investigated PCR-clamping efficiency by studying PNA position and mismatch with complementary DNA by designing PNAs at five different positions on the nuclear rDNA internal transcribed spacer 1 of the Japanese eel Anguilla japonica in association with intra-specific nucleotide substitutions. All five PNAs were observed to efficiently inhibit amplification of a fully complementary DNA template. One mismatch between PNA and template DNA inhibited amplification of the template DNA, while two or more mismatches did not. DNA samples extracted from dorsal muscle and intestine of eight wild-caught leptochephalus larvae were subjected to this analysis, followed by cloning, nucleotide sequence analysis, and database homology search. Among 12 sequence types obtained from the intestine sample, six were identified as fungi. No sequence similarities were found in the database for the remaining six types, which were not related to one another. These results, in conjunction with our laboratory observations on larval feeding, suggest that eel leptocephali may not be dependent upon living plankton for their food source.


Narrow-linewidth operation of folded 1178nm VECSEL with twisted-mode cavity.
A vertical external-cavity surface-emitting laser (VECSEL) with a twisted-mode configuration is demonstrated. This architecture is particularly advantageous for power scaling of single-frequency VECSELs employing multiple gain mirrors in folded cavities. In such a configuration, some of the gain mirrors are inherently at the fold, and the lasing spectrum becomes unstable. This is caused by four waves interfering, destabilizing the standing wave pattern at the quantum wells. We show that the lasing spectrum can be narrowed by employing a twisted-mode configuration, which stabilizes the standing-wave pattern at the gain mirror. Furthermore, single-frequency output of more than 10 W at 1178 nm is demonstrated for a VECSEL employing two gain mirrors in a standing-wave cavity. In comparison, the output power for operation with one gain mirror only was 7.4 W when operating in single frequency. The choice of wavelength for the work reported in this paper is motivated by the opportunity to demonstrate compact VECSEL-based guide star lasers for adaptive optics via frequency doubling to the sodium D2 resonance at 589 nm.


Increased expression of ceruloplasmin in the retina following photic injury.
Oxidative stress plays a role in the photic injury model of retinal degeneration and in age-related macular degeneration. Our preliminary microarray analysis of retinal gene expression upon photic injury suggested increased expression of ceruloplasmin, a ferroxidase that could reduce retinal oxidative stress. Patients with acerul oplasminemia have retinal degeneration, indicating that ceruloplasmin is necessary for maintenance of retinal health. The purpose of this study was to determine whether retinal ceruloplasmin is upregulated following photo-oxidation, to localize ceruloplasmin protein, and to determine which ceruloplasmin isoform is present in the retina. Balb/c mice were exposed to bright white light for seven hours. TUNEL labeling was used to detect photoreceptor apoptosis. At several intervals after the light injury, retinal ceruloplasmin was studied by quantitative PCR, immunohistochemistry, and western analysis. Expression of the secreted and expression of the membrane-anchored glycosyl phosphatidyl inositol (GPI) linked forms of ceruloplasmin were assesed in rat retina using primers specific for each form. Vitreous ceruloplasmin was detected by immunohistochemistry in Balb/c mouse eyes and by western analysis of aspirated vitreous from post-mortem human eyes. Retinal ceruloplasmin mRNA was upregulated eight-fold following photic injury. Ceruloplasmin protein was detected throughout normal retinas by immunohistochemistry, with a specific increase in Muller cell labeling following photic injury. Western analysis confirmed an increase in ceruloplasmin protein following photic injury and revealed eight-fold more ceruloplasmin protein in normal retina than in brain. The mRNAs for both the secreted and GPI linked forms of ceruloplasmin were detected by RT-PCR in the retina. Ceruloplasmin protein was detected by western analysis of normal human vitreous and was increased in mouse vitreous following photic injury. Ceruloplasmin, a retinal ferroxidase, is upregulated at the mRNA and protein levels upon light damage. The increased protein is primarily in Muller cells. Ceruloplasmin is considerably more abundant in retina than in brain. The retina expresses both the GPI-linked and secreted forms of ceruloplasmin, and since vitreous ceruloplasmin increases following photic injury, some of the retinal ceruloplasmin may be secreted into the vitreous. Ceruloplasmin may protect the retina from oxidative stress by decreasing the amount of ferrous iron available to produce reactive oxygen species.