The mechanical waveform of the basilar membrane. IV. Tone and noise stimuli.
Analysis of mechanical cochlear responses to wide bands of random noise clarifies many effects of cochlear nonlinearity. The previous paper [de Boer and Nuttall, J. Acoust. Soc. Am. 107, 1497-1507 (2000)] illustrates how closely results of computations in a nonlinear cochlear model agree with responses from physiological experiments. In the present paper results for tone stimuli are reported. It was found that the measured frequency response for pure tones differs little from the frequency response associated with a noise signal. For strong stimuli, well into the nonlinear region, tones have to be presented at a specific level with respect to the noise for this to be true. In this report the nonlinear cochlear model originally developed for noise analysis was modified to accommodate pure tones. For this purpose the efficiency with which outer hair cells modify the basilar-membrane response was made into a function of cochlear location based on local excitation. For each experiment, the modified model is able to account for the experimental findings, within 1 or 2 dB. Therefore, the model explains why the type of filtering that tones undergo in the cochlea is essentially the same as that for noise signals (provided the tones are presented at the appropriate level).


Neurogenic and myogenic patterns of electrical activity in isolated intact mouse colon.
Relatively little is known about the electrical rhythmicity of the whole colon, where long neural pathways are preserved. Smooth muscle electrical activity was recorded extracellularly from the serosa of isolated flat-sheet preparations consisting of the whole mouse colon (n=31). Two distinct electrical patterns were observed. The first, long intense spike bursts, occurred every 349±256 seconds (0.2±0.2 cpm), firing action potentials for 31±11 seconds at 2.1±0.5 Hz. They were hexamethonium- and tetrodotoxin-sensitive, but persisted in nicardipine as 2 Hz electrical oscillations lacking action potentials. This pattern is called here neurogenic spike bursts. The second pattern, short spike bursts, occurred about every 30 seconds (2.0±0.6 cpm), with action potentials firing at about 1 Hz for 9 seconds (1.0±0.2 Hz, 9±4 seconds). Short spike bursts were hexamethonium- and tetrodotoxin-resistant but nicardipine-sensitive and thus called here myogenic spike bursts. Neurogenic spike bursts transiently delayed myogenic spike bursts, while blocking neurogenic activity enhanced myogenic spike burst durations. External stimuli significantly affected neurogenic but not myogenic spike bursts. Aboral electrical or mechanical stimuli evoked premature neurogenic spike bursts. Circumferential stretch significantly decreased intervals between neurogenic spike bursts. Lesioning the colon down to 10 mm segments significantly increased intervals or abolished neurogenic spike bursts, while myogenic spike bursts persisted. Distinct neurogenic and myogenic electrical patterns were recorded from mouse colonic muscularis externa. Neurogenic spike bursts likely correlate with neurogenic colonic migrating motor complexes (CMMC) and are highly sensitive to mechanical stimuli. Myogenic spike bursts may correspond to slow myogenic contractions, whose duration can be modulated by enteric neural activity.


Extended lactation in high-yielding dairy cows. I. Effects on reproductive measurements.
The objective of this prospective field study was to evaluate the effects of extending the lactation period on various reproductive measurements of high-yielding Holstein cows. On 40 d in milk (DIM), cows were gynecologically examined (transrectal palpation, sonography, vaginoscopy). Cows without signs of clinical endometritis were blocked by parity and were randomly allocated to 1 of 3 experimental groups with a voluntary waiting period (VWP) of 40, 120, and 180 d, respectively (G40, n = 135; G120, n = 141; G180, n = 139). Cows of G120 and G180 were reexamined at the end of the VWP. If natural estrus was detected within 46 d after the end of the VWP, an artificial insemination was performed. If no estrus was detected, the respective cows were synchronized by applying the classical Ovsynch protocol. We found no difference in the proportion of cows in which estrus was detected between 40 to 86 DIM or in the days to first estrus between the 3 groups. Estrus detection in this period was lower in cows with body condition score <3 on 90 DIM compared with body condition score ≥3 (61.5 vs. 76.0%) and in cows with high energy-corrected milk production (ECM) on 92 DIM [58.6 vs. 70.1%, for cows with higher and lower than the median (39.9 kg) ECM, respectively]. The proportion of cows that estrus was detected within 46 d after the VWP was greater in G120 (88.9%) and G180 (90.8%) compared with G40 (70.4%). These effects were more apparent in cows with high ECM. The rate of estrus detection and of becoming pregnant in this period was greater for G120 (hazard ratio = 2.2 and 1.6, respectively) and for G180 (hazard ratio = 2.4 and 1.8) compared with G40. Cows in both groups with extended lactation had greater overall first service conception rates (G120 = 48.9%; G180 = 49.6%) and a lower number of services per pregnant cow (G120 = 1.56 ± 0.1; G180 = 1.51 ± 0.1) compared with G40 (36.6%; 1.77 ± 0.1). We observed no difference in pregnancy loss or in the proportion of cows culled up to 305 d of lactation between the 3 groups. The number of Ovsynch protocols per 1,000,000 kg of ECM was reduced by 75% in G180 and by 74% in G120 compared with G40 (5.9 vs. 7.1 vs. 25.1). In conclusion, extending the lactation of dairy cows can improve main reproductive measurements in high-yielding cows.


Radial width of the temporal horn: a sensitive measure in Alzheimer disease.
Atrophy in the medial temporal lobe (MTL) structures depicted with brain imaging is one of the most accurate markers of Alzheimer disease (AD), but practical considerations have thus far limited their routine clinical use. The aim of this study was to explore the validity of a CT- and MR-based measure of MTL atrophy that would be feasible for routine clinical use. We acquired brain CT scans in the temporal lobe plane with thin sections in 42 patients with AD and in 29 control patients without dementia. We also acquired MR images (according to a 3D magnetization-prepared rapid gradient-echo protocol) in 28 patients with AD and in 28 control subjects without dementia. The radial width of the temporal horn (rWTH) of the lateral ventricle was measured with a precision caliper at the tip of the horn on CT scans or high-quality MR images. The validity of the rWTH variable was assessed by test-retest and interrater reliability, convergent and discriminant validity compared with progressively distant brain regions, and known-group validity (accuracy of the separation of patients with AD from control subjects). Convergent and discriminant validity compared with volumetric measures was tested in the patients who underwent MR imaging. Intraclass correlation coefficients for inter- and intrarater reliability were between 0.94 and 0.98. On CT scans, Pearson's correlation of the rWTH with the transverse width of the temporal horn was between 0.60 and 0.79; with Jobst's minimum thickness of the MTL, between 0.63 and 0.78 (interuncal distance approximately 0.50); and with an index of frontal atrophy, between 0.35 and 0.42. On MR images, the correlation with volumetric MR measures was 0.80 in the temporal horn, 0.74 in the hippocampus, 0.68 in the temporal lobe, 0.58 in the entorhinal cortex, and 0.49 in the frontal lobe. On CT scans (cutoff value for AD, >5.3 mm; age range of subjects, 50-90 y), the rWTH measure was a sensitive marker for AD in 39 of 42 patients with AD (sensitivity, 93%) and was a specific marker in 28 of the 29 control patients (specificity, 97%). On MR images (cutoff 3.6-6.7 mm; age range of subject, 50-90 y), the rWTH was a sensitive marker for AD in 21 of 28 patients with AD (sensitivity, 75%) and was a specific marker in 26 of 28 control subjects (specificity, 93%). The accuracy of other linear CT-based measures of MTL atrophy and linear and volumetric MR-based measures was lower. With specificity set to 95%, sensitivity ranged from 57% to 74% for CT-based measures and from 52% to 74% for MR-based measures. The rWTH is an accurate marker of AD that could be used in routine clinical settings.


General patchy ellipsoidal particle model for the aggregation behaviors of shape- and/or surface-anisotropic building blocks.
We present a general patchy ellipsoidal particle model suitable for conducting dynamics simulations of the aggregation behaviors of various shape- and/or surface-anisotropic colloids, especially patchy ellipsoids with continuously variable shape and tunable patchiness. To achieve higher computational efficiency in dynamics simulations, we employ a multi-GPU acceleration technique based on a domain decomposition algorithm. The validation and performance evaluation of this GPU-assisted model are performed by simulating several typical benchmark systems of non-patchy and patchy ellipsoids. Given the generality and efficiency of our GPU-assisted patchy ellipsoidal particle model, it will provide a highly feasible dynamics simulation framework to investigate the aggregation behaviors of anisotropic soft matter systems comprised of shape- and/or surface-anisotropic building blocks.


Bifunctional catalyst promotes highly enantioselective bromolactonizations to generate stereogenic C-Br bonds.
A novel bifunctional catalyst derived from BINOL has been developed that promotes the highly enantioselective bromolactonizations of a number of structurally distinct unsaturated acids. Like some known catalysts, this catalyst promotes highly enantioselective bromolactonizations of 4- and 5-aryl-4-pentenoic acids, but it also catalyzes the highly enantioselective bromolactonizations of 5-alkyl-4(Z)-pentenoic acids. These reactions represent the first catalytic bromolactonizations of alkyl-substituted olefinic acids that proceed via 5-exo mode cyclizations to give lactones in which new carbon-bromine bonds are formed at a stereogenic center with high enantioselectivity. We also disclose the first catalytic desymmetrization of a prochiral dienoic acid by enantioselective bromolactonization.


Freely distributed bed-net use among Chano Mille residents, south Ethiopia: a longitudinal study.
A huge discrepancy was reported between ownership versus utilization of insecticide-treated bed nets (ITNs). To acquire the benefits of ITNs, households need to use and not merely own them. The objective of this study was to characterize the pattern of, and assess factors related to ITN use in one village in south Ethiopia. A prospective cohort study involving 8,121 residents (in 1,388 households) was carried out from April 2009 to April 2011 (101 weeks). Every week, individuals were asked whether they slept under an ITN the night before the interview. Descriptive statistics was used to report the availability and use of ITN. A negative, binomial, probability, distribution model was fitted to find out significant predictors of ITN use. Reasons for not using ITN were summarized. The total number of ITNs available at the beginning of the study was 1,631 (1.68 ITNs per household). On week 48, 3,099 new ITNs (PermaNet2.0) were distributed freely (2.3 ITNs per household). The number of households who received at least one new ITN was 1,309 (98.4%). The percentage of children <5 years and pregnant women not using ITNs exceeded that of other adults. The mean (range; SD) ITN use fraction before and after mass distribution was 0.20 (0.15-0.27; 0.03) and 0.62 (0.47-0.69; 0.04), respectively. Before mass ITN distribution, the most frequent reason for not using ITN was having worn out bed nets (most complained the bed nets were torn by rats); and after mass ITN distribution, it was lack of convenient space to hang more than one ITN. Males, younger age groups (mainly 15-24 years) and those living away from the vector-breeding site were less likely to use ITN. The ITN use fraction reached to a maximum of 69% despite near universal coverage (98.4%) was achieved. Gender, age differences and distance from vector breeding site were associated with ITN use. Strategies may need to be designed addressing disproportions in ITN use, lack of convenient space to hang more than one ITN (for those receiving more than one), and measures to prolong usable life of ITNs.


The adaptor protein p62/SQSTM1 in osteoclast signaling pathways.
Paget's disease of bone (PDB) is a skeletal disorder characterized by focal and disorganized increases in bone turnover and overactive osteoclasts. The discovery of mutations in the SQSTM1/p62 gene in numerous patients has identified protein p62 as an important modulator of bone turnover. In both precursors and mature osteoclasts, the interaction between receptor activator of NF-κB ligand (RANKL) and its receptor RANK results in signaling cascades that ultimately activate transcription factors, particularly NF-κB and NFATc1, promoting and regulating the osteoclast differentiation, activity, and survival. As a scaffold with multiple protein-protein interaction motifs, p62 is involved in virtually all the RANKL-activated osteoclast signaling pathways, along with being implicated in numerous other cellular processes. The p62 adaptor protein is one of the functional links reported between RANKL and TRAF6-mediated NF-κB activation, and also plays a major role as a shuttling factor that targets polyubiquitinated proteins for degradation by either the autophagy or proteasome pathways. The dysregulated expression and/or activity of p62 in bone disease up-regulates osteoclast functions. This review aims to outline and summarize the role of p62 in RANKL-induced signaling pathways and in ubiquitin-mediated signaling in osteoclasts, and the impact of PDB-associated p62 mutations on these processes.


Bioassay of chloramben for possible carcinogenicity.
A bioassay of technical-grade chloramben for possible carcinogenicity was conducted by administering the test material in feed to Osborne-Mendel rats and B6C3F1 mice. Groups of 50 rats and 50 mice of both sexes were administered chloramben at one of two doses, either 10,000 or 20,000 ppm. The rats were treated for 80 weeks, then observed for 32 or 33 weeks; the mice were treated for 80 weeks, then observed for 11 or 12 weeks. Matched controls consisted of groups of 10 untreated rats and 10 untreated mice of each sex; pooled controls, used for statistical evaluation, consisted of these matched controls combined with 75 untreated male and 75 untreated female rats or 70 untreated male and 70 untreated female mice from similarly performed bioassays of six other test chemicals. Surviving rats were killed at 112 or 113 weeks; surviving mice were killed at 91 or 92 weeks. Body weights and mortality of the treated animals were not markedly affected by chloramben under the conditions of the bioassay. The various clinical signs observed were common to both treated and control groups. In male rats, hemangiomas occurred at a significantly higher incidence in the low-dose animals than in the pooled controls (controls 0/73, low-dose 5/48, P=0.009). This lesion was not considered to be related to the administration of chloramben, since the tumor did not occur at a significantly higher incidence in the high-dose group than in the pooled-control group, and the incidences did not show a significant dose-related trend. In both male and female mice, the incidences of hepatocellular carcinoma showed significant dose-related trends using pooled controls (for females: controls 9/69, low dose 16/48, high-dose 14/48, P=0.029; for females controls 2/67, low-dose 7/48, high-dose 10/50, P=0.004). Direct comparisons showed significantly higher incidences of the tumor in the low-dose males (P=0.008) and in the high-dose females (P=0.003) than in the pooled controls. Probability levels of P=0.028 in high-dose males and P=0.027 in low-dose females were attained. In male mice, however, the incidence of hepatocellular carcinoma was considered to be only marginally associated with the administration of chloramben because of the variations in the spontaneous incidence of this lesion in male mice encountered at this laboratory. In conclusion, under the conditions of this bioassay, there were no tumors in Osborne-Mendel rats that were significantly related to administration of the chemical. In B6C3F1 female mice, chloramben was carcinogenic, producing hepatocellular carcinomas in treated animals.


Metabolic Syndrome and Symptom Resolution in Depression: A 5-Year Follow-Up of Older Adults.
Although metabolic syndrome is associated with the incidence of depression, little is known about its contribution to the course of depression. We examined whether metabolic syndrome and its components are associated with long-term symptom resolution in older adults with depressive symptoms. Data from 965 participants in the Whitehall II cohort study (mean age = 62 years at baseline) were used to generate 1,172 person-observations of metabolic syndrome and its components (abdominal obesity, low level of high-density lipoprotein [HDL] cholesterol, high level of triglycerides, hypertension, and elevated fasting glucose or diabetes). All participants were depression cases at the beginning of 2 consecutive follow-up cycles: from 2002-2004 to 2007-2009 and from 2007-2009 to 2012-2013 (mean follow-up = 4.6 years). Depressive symptoms were assessed using the Center for Epidemiologic Studies Depression scale caseness at the beginning and the end of the 2 cycles. In multivariable adjusted analyses, metabolic syndrome per se was not associated with symptom resolution. Of its components, low HDL cholesterol (risk ratio [RR] = 0.82; 95% CI, 0.68-1.00; P = .045) and high triglyceride levels (RR = 0.81; 95% CI, 0.70-0.95; P = .007) were associated with a lower likelihood of symptom resolution. These findings were replicated in a subpopulation without coronary heart disease and stroke (RR = 0.77 [95% CI, 0.63-0.95; P = .015] for low HDL cholesterol; RR = 0.79 [95% CI, 0.67-0.94; P = .006] for high triglycerides). Low HDL cholesterol and high triglyceride levels are associated with lower likelihood of long-term symptom resolution in depression. These data suggest that an adverse lipid profile, but not other components of metabolic syndrome, may delay recovery from depression.


Interaction between arsenic exposure from drinking water and genetic susceptibility in carotid intima-media thickness in Bangladesh.
Epidemiologic studies that evaluated genetic susceptibility for the effects of arsenic exposure from drinking water on subclinical atherosclerosis are limited. We conducted a cross-sectional study of 1078 participants randomly selected from the Health Effects of Arsenic Longitudinal Study in Bangladesh to evaluate whether the association between arsenic exposure and carotid artery intima-media thickness (cIMT) differs by 207 single-nucleotide polymorphisms (SNPs) in 18 genes related to arsenic metabolism, oxidative stress, inflammation, and endothelial dysfunction. Although not statistically significant after correcting for multiple testing, nine SNPs in APOE, AS3MT, PNP, and TNF genes had a nominally statistically significant interaction with well-water arsenic in cIMT. For instance, the joint presence of a higher level of well-water arsenic (≥ 40.4 μg/L) and the GG genotype of AS3MT rs3740392 was associated with a difference of 40.9 μm (95% CI = 14.4, 67.5) in cIMT, much greater than the difference of cIMT associated with the genotype alone (β = -5.1 μm, 95% CI = -31.6, 21.3) or arsenic exposure alone (β = 7.2 μm, 95% CI = -3.1, 17.5). The pattern and magnitude of the interactions were similar when urinary arsenic was used as the exposure variable. Additionally, the at-risk genotypes of the AS3MT SNPs were positively related to the proportion of monomethylarsonic acid (MMA) in urine, which is indicative of arsenic methylation capacity. The findings provide novel evidence that genetic variants related to arsenic metabolism may play an important role in arsenic-induced subclinical atherosclerosis. Future replication studies in diverse populations are needed to confirm the findings.


A Drosophila Model of Essential Tremor.
Essential Tremor (ET) is one of the most common neurological diseases, with an estimated 7 million affected individuals in the US; the pathophysiology of the disorder is poorly understood. Recently, we identified a mutation (KCNS2 (Kv9.2), c.1137 T > A, p.(D379E) in an electrically silent voltage-gated K+ channel α-subunit, Kv9.2, in a family with ET, that modulates the activity of Kv2 channels. We have produced transgenic Drosophila lines that express either the human wild type Kv9.2 (hKv9.2) or the ET causing mutant Kv9.2 (hKv9.2-D379E) subunit in all neurons. We show that the hKv9.2 subunit modulates activity of endogenous Drosophila K+ channel Shab. The mutant hKv9.2-D379E subunit showed significantly higher levels of Shab inactivation and a higher frequency of spontaneous firing rate consistent with neuronal hyperexcitibility. We also observed behavioral manifestations of nervous system dysfunction including effects on night time activity and sleep. This functional data further supports the pathogenicity of the KCNS2 (p.D379E) mutation, consistent with our prior observations including co-segregation with ET in a family, a likely pathogenic change in the channel pore domain and absence from population databases. The Drosophila hKv9.2 transgenic model recapitulates several features of ET and may be employed to advance our understanding of ET disease pathogenesis.


Does physiologic response to loud tones change following cognitive-behavioral treatment for posttraumatic stress disorder?
This study examined responses to loud tones before and after cognitive-behavioral treatment for posttraumatic stress disorder (PTSD). Seventy-four women in a PTSD treatment outcome study for rape-related (n = 54) or physical assault-related PTSD (n = 20) were assessed in an auditory loud tone paradigm. Assessments were conducted before and after a 6-week period of cognitive-behavioral therapy. Physiologic responses to loud tones included heart rate (HR), skin conductance (SC), and eye-blink electromyogram (EMG). Groups were formed based upon treatment outcome and included a treatment responder group (no PTSD at posttreatment) and a nonresponder group (PTSD-positive at posttreatment). Treatment was successful for 53 of 74 women (72%) and unsuccessful for 21 women (28%). Responders and nonresponders were not significantly different from each other at pretreatment on the main outcome variables. Treatment responders showed a significant reduction in loud tone-related EMG, HR, and SC responses from pre- to posttreatment (partial η(2) = .24, .31, and .36, respectively; all p < .001) and the EMG and HR responses were significantly smaller than nonresponders at posttreatment (partial η(2) = .11, p = .004 and .19, p < .001, respectively). Successful cognitive-behavioral treatment of PTSD is associated with a quantifiable reduction in physiological responding to loud tones.


The Relationship between Ethnic Classroom Composition and Turkish-Origin and German Students' Reading Performance and Sense of Belonging.
Past research on ethnic composition effects on migrant and ethnic majority students' performance has reported inconclusive results: Some studies have found no relationship between the proportion of migrant students in school and students' performance, some revealed positive effects, whereas others showed negative effects of the proportion of migrant students. Most of the studies did not consider whether an increase in the proportion of migrant students in the classroom has different effects on migrant and ethnic majority students' performance. For this reason, the present study (N = 9215) extends previous research by investigating the cross-level interaction effect of the proportion of Turkish-origin students in classrooms on Turkish-origin and German students' reading performance with data based on the German National Assessment Study 2008/2009 in the school subject German. In addition, we examined the cross-level interaction effect of Turkish-origin students' proportion on sense of belonging to school for Turkish-origin and German students, as sense of belonging has been shown to be an important predictor of well-being and integration. No cross-level interaction effect on performance emerged. Only a small negative main effect of the Turkish-origin students' proportion on all students' performance was found. As predicted, we showed a cross-level interaction on sense of belonging. Only Turkish-origin students' sense of belonging was positively related to the proportion of Turkish-origin students: The more Turkish-origin students there were in a classroom, the higher Turkish-origin students' sense of belonging. German students' sense of belonging was not related to the ethnic classroom composition. Implications of the results in the educational context are discussed.


Chin surgery: I. Augmentation--the allures and the alerts.
The correction of sagittal deformities of the chin presents a seemingly simple surgical challenge. However, several authors have reported negative sequelae from such chin surgery, During the past 11 years, the senior author (B.M.Z.) has evaluated more than 100 such cases of adverse results after chin augmentation. Many surgeons, it seems, use chin implants unnecessarily and, thus, get into trouble. Because alloplastic chin augmentation is deceptively easy, it tends to be overused in certain situations. Either the surgeon's evaluation is too narrowly focused or his/her abilities to perform other types of surgery (e.g., osseous genioplasty) are limited. Herein, the authors present a diagnostic evaluation protocol, QUAC (Quick Analysis of the Chin), to assist in avoiding simple mistakes in alloplastic chin augmentation. This protocol will alert the surgeon to situations that, if unrecognized, will cause problems and create an unhappy patient. This article will specifically focus on (1) lower lip analysis; (2) the effect of the labiomental fold; (3) chin pad evaluation, both static and dynamic; (4) the anatomy of the cleft chin; (5) special situations; and (6) how to troubleshoot three common problems. The accompanying article, Chin Surgery II, will present a new operation that treats a chin problem that was previously difficult to correct.


Characterization and sleep deprivation-induced expression modulation of dendrin, a novel dendritic protein in rat brain neurons.
We report on the characterization of the novel rat brain protein dendrin which is encoded by the brain-specific transcript 464. On immunoblots, two protein variants (81 kD, 89 kD) were identified in cytosolic and membraneous protein fractions. The variants are most abundant in the hippocampus, notably in apical dendrites of CA1 pyramidal cells. Dendritic and perikaryal immunolabelling is apparent in neurons of the cerebral cortex, dentate gyrus, subiculum, amygdala, and preoptic areas. In cortical and hippocampal dendrites, electron-dense immunoreaction is associated with the endoplasmic reticulum, the plasma membrane, and spine heads. An association of dendrin with polyribosomes and the presence of its mRNA in dendrites both provide evidence for dendritic mRNA translation. In the rat forebrain, dendrin expression is altered after an extended period of wakefulness. Twenty-four-hour sleep deprivation decreases the mRNA and protein concentrations of both variants in subcortical forebrain plus midbrain areas by 24 +/- 11% (P < 0.05) and 40 +/- 14% (P < 0.1), respectively, as measured relative to beta-actin mRNA and neural actin. In the cerebral cortex and hippocampus, the relative mRNA level remains unchanged whereas the cortical protein concentration is reduced by 42 +/- 10% (P < 0.05). Thus, dendrin belongs to a new class of dendritic proteins whose expression is differentially modulated by prolonged behavioral activity.


Structural and functional study of D-glucuronyl C5-epimerase.
Heparan sulfate (HS) is a glycosaminoglycan present on the cell surface and in the extracellular matrix, which interacts with diverse signal molecules and is essential for many physiological processes including embryonic development, cell growth, inflammation, and blood coagulation. D-glucuronyl C5-epimerase (Glce) is a crucial enzyme in HS synthesis, converting D-glucuronic acid to L-iduronic acid to increase HS flexibility. This modification of HS is important for protein ligand recognition. We have determined the crystal structures of Glce in apo-form (unliganded) and in complex with heparin hexasaccharide (product of Glce following O-sulfation), both in a stable dimer conformation. A Glce dimer contains two catalytic sites, each at a positively charged cleft in C-terminal α-helical domains binding one negatively charged hexasaccharide. Based on the structural and mutagenesis studies, three tyrosine residues, Tyr(468), Tyr(528), and Tyr(546), in the active site were found to be crucial for the enzymatic activity. The complex structure also reveals the mechanism of product inhibition (i.e. 2-O- and 6-O-sulfation of HS keeps the C5 carbon of L-iduronic acid away from the active-site tyrosine residues). Our structural and functional data advance understanding of the key modification in HS biosynthesis.


Training generalist doctors for rural practice in New Zealand.
Targeted postgraduate training increases the likelihood young doctors will take up careers in rural generalist medicine. This article describes the postgraduate pathways that have evolved for these doctors in New Zealand. The Cairns consensus statement 2014 defined rural medical generalism as a scope of practice that encompasses primary care, hospital or secondary care, emergency care, advanced skill sets and a population-based approach to the health needs of rural communities. Even as work goes on to define this role different jurisdictions have developed their own training pathways for these important members of the rural healthcare workforce. In 2002 the University of Otago developed a distance-taught postgraduate diploma aimed at the extended practice of rural general practitioners (GPs) and rural hospital medical officers. This qualification has evolved into a 4-year vocational training program in rural hospital medicine, with the university diploma retained as the academic component. The intentionally flexible and modular nature of the rural hospital training program and university diploma allow for a range of training options. The majority of trainees are taking advantage of this by combining general practice and rural hospital training. Although structured quite differently the components of this combined pathway looks similar to the Australian rural generalist pathways. There is evidence that the program has had a positive impact on the New Zealand rural hospital medical workforce.


Improving Blood Pressure Management in Primary Care Patients with Chronic Kidney Disease: a Systematic Review of Interventions and Implementation Strategies.
Chronic kidney disease (CKD) is widely prevalent, associated with morbidity and mortality, but may be lessened with timely implementation of evidence-based strategies including blood pressure (BP) control. Nonetheless, an evidence-practice gap persists. We synthesize the evidence for clinician-facing interventions to improve hypertension management in CKD patients in primary care. Electronic databases and related publications were queried for relevant studies. We used a conceptual model to address heterogeneity of interventions. We conducted a quantitative synthesis of interventions on blood pressure (BP) outcomes and a narrative synthesis of other CKD relevant clinical outcomes. Planned subgroup analyses were performed by (1) study design (randomized controlled trials (RCTs) or nonrandomized studies (NRS)); (2) intervention type (guideline-concordant decision support, shared care, pharmacist-facing); and (3) use of behavioral/implementation theory. Of 2704 manuscripts screened, 73 underwent full-text review; 22 met inclusion criteria. BP target achievement was reported in 15 and systolic BP reduction in 6 studies. Among RCTs, all interventions had a significant effect on BP control, (pooled OR 1.21; 95% CI 1.07 to 1.38). Subgroup analysis by intervention type showed significant effects for guideline-concordant decision support (pooled OR 1.19; 95% CI 1.12 to 1.27) but not shared care (pooled OR 1.71; 95% CI 0.96 to 3.03) or pharmacist-facing interventions (pooled OR 1.04; 95% CI 0.82 to 1.34). Subgroup analysis finding was replicated with pooling of RCTs and NRS. The five contributing studies showed large and significant reduction in systolic BP (pooled WMD - 3.86; 95% CI - 7.2 to - 0.55). Use of a behavioral/implementation theory had no impact, while RCTs showed smaller effect sizes than NRS. Process-oriented implementation strategies used with guideline-concordant decision support was a promising implementation approach. Better reporting guidelines on implementation would enable more useful synthesis of the efficacy of CKD clinical interventions integrated into primary care. CRD42018102441.


An Effective Peptide-Based Platform for Efficient Exosomal Loading and Cellular Delivery of a microRNA.
Exosomes, membrane-bound nanosized vesicles of biologic origin, are known to contain various molecules, e.g., proteins, lipids, and nucleic acids, which contribute to the exosomes' ability to mediate cell-to-cell communication. Recent impediments of artificial nanoparticles in drug delivery, including low cellular uptake, activation of the immune system, and tissue obstacles, have led scientists to engineer exosomes as drug delivery vehicles. Though exosomes possess inherent properties of stability, biocompatibility, low immunogenicity, and capability to cross biological barriers, there is a need to develop technologies that allow the efficient loading of therapeutic materials into exosomes. Here, we introduced a simple peptide-equipped technology that can enhance the cargo-loading potential of exosomes in a mild loading environment. Specifically, a known cell-penetrating peptide, YARA, derived from human immunodeficiency virus-1 trans-activator of transcription, was covalently conjugated with miR-21-5p, a mammalian microRNA. The conjugate YARA-miR-21-5p was then incubated with exosomes, isolated from either mesenchymal stem cells or cancer cells, for loading. Exosomal loading of YARA-miR-21-5p was time-dependent and demonstrated an impressive 18.6-fold increase in efficiency over exosomal loading of miR-21-5p through incubation. After effective cellular uptake, the loaded exosomes rapidly delivered YARA-miR-21-5p into mammalian cells. Relative to unloaded exosomes and free YARA-miR-21-5p, the loaded exosomes significantly enhanced the proliferation, migration, and invasion of human and mouse fibroblasts, which are vital steps in wound healing. This study lays the groundwork for using cell-penetrating peptides as an innovative approach to efficiently load therapeutic cargos, e.g., microRNAs, into exosomes, which can then be employed to deliver the cargos into cells to yield biological effects.


Alkyl triphenylphosphonium surfactants as nucleic acid carriers: complexation efficacy toward DNA decamers, interaction with lipid bilayers and cytotoxicity studies.
Herein, for the first time the complexation ability of a homological series of triphenylphosphonium surfactants (TPPB-n) toward DNA decamers has been explored. Formation of lipoplexes was confirmed by alternative techniques, including dynamic light scattering, indicating the occurrence of nanosized complexes (ca. 100-150 nm), and monitoring the charge neutralization of nucleotide phosphate groups and the fluorescence quenching of dye-intercalator ethidium bromide. The complexation efficacy of TPPB-surfactants toward an oligonucleotide (ONu) is compared with that of reference cationic surfactants. Strong effects of the alkyl chain length and the structure of the head group on the surfactant/ONu interaction are revealed, which probably occur via different mechanisms, with electrostatic and hydrophobic forces or intercalation imbedding involved. Phosphonium surfactants are shown to be capable of disordering lipid bilayers, which is supported by a decrease in the temperature of the main phase transition, Tm. This effect enhances with an increase in the alkyl chain length, indicating the integration of TPPB-n with lipid membranes. This markedly differs from the behavior of typical cationic surfactant cetyltrimethylammonium bromide, which induces an increase in the Tm value. It was demonstrated that the cytotoxicity of TPPB-n in terms of the MTT-test on a human cell line 293T nonmonotonically changes within the homological series, with the highest cytotoxicity exhibited by the dodecyl and tetradecyl homologs.


Characteristics of health information gatherers, disseminators, and blockers within families at risk of hereditary cancer: implications for family health communication interventions.
Given the importance of the dissemination of accurate family history to assess disease risk, we characterized the gatherers, disseminators, and blockers of health information within families at high genetic risk of cancer. A total of 5466 personal network members of 183 female participants of the Breast Imaging Study from 124 families with known mutations in the BRCA1/2 genes (associated with high risk of breast, ovarian, and other types of cancer) were identified by using the Colored Eco-Genetic Relationship Map (CEGRM). Hierarchical nonlinear models were fitted to characterize information gatherers, disseminators, and blockers. Gatherers of information were more often female (P<.001), parents (P<.001), and emotional support providers (P<.001). Disseminators were more likely female first- and second-degree relatives (both P<.001), family members in the older or same generation as the participant (P<.001), those with a cancer history (P<.001), and providers of emotional (P<.001) or tangible support (P<.001). Blockers tended to be spouses or partners (P<.001) and male, first-degree relatives (P<.001). Our results provide insight into which family members may, within a family-based intervention, effectively gather family risk information, disseminate information, and encourage discussions regarding shared family risk.


A rare cause of mental motor retardation: recessive congenital methemoglobinemia type II.
Recessive congenital methemoglobinemia (RCM) is a very rare disorder caused by NADH- cytochrome b5 reductase (cytb5r) deficiency. It has been classified into four types. Type I presents with mild cyanosis due to a significant deficiency of cytb5r in erythrocytes only. In type II, the deficiency occurs in all tissues and causes growth and mental retardation and other neurological impairments. RCM types I and II are caused by a defect in a single gene, which is located on chromosome 22 (locus DIA 1: q 13.31-qter). Prenatal diagnosis is possible. Cyanosis can be well treated by 200-500 mg of ascorbic acid daily; there is no effective therapy for the progressive neurological impairments. This report presents two siblings with central cyanosis, growth retardation, mental retardation, microcephaly, dystonia and hypertonia diagnosed as RCM type II.


Constructing gene association networks for rheumatoid arthritis using the backward genotype-trait association (BGTA) algorithm.
Rheumatoid arthritis (RA, MIM 180300) is a common and complex inflammatory disorder. The North American Rheumatoid Arthritis Consortium (NARAC) data, as part of the Genetic Analysis Workshop 15 data, consists of both genome scan and candidate gene studies on RA patients. We applied the backward genotype-trait association (BGTA) algorithm to capture marginal and gene x gene interaction effects of multiple susceptibility loci on RA disease status. A two-stage screening approach was used for the genome scan, whereas a comprehensive study of all possible subsets was conducted for the candidate genes. For the genome scan, we constructed an association network among 39 genetic loci that demonstrated strong signals, 19 of which have been reported in the RA literature. For the candidate genes, we found strong signals for PTPN22 and SUMO4. Based on significant association evidence, we built an association network among the loci of PTPN22, PADI4, DLG5, SLC22A4, SUMO4, and CARD15. To control for false positives, we used permutation tests to constrain the family-wise type I error rate to 1%. Using the BGTA algorithm, we identified genetic loci and candidate genes that were associated with RA susceptibility and association networks among them. For the first time, we report possible interactions between single-nucleotide polymorphisms/genes, which may be useful for biological interpretation.


The Potential for Decision Support Tools to Improve the Management of Root-Feeding Fly Pests of Vegetables in Western Europe.
Several important vegetable crops grown outdoors in temperate climates in Europe can be damaged by the root-feeding larvae of Diptera (Delia radicum, Delia floralis, Chamaepsila rosae, Delia platura, Delia florilega, Delia antiqua). Knowledge of pest insect phenology is a key component of any Integrated Pest Management (IPM) strategy, and this review considers the methods used to monitor and forecast the occurrence of root-feeding flies as a basis for decision-making by growers and the ways that such information can be applied. It has highlighted some current management approaches where such information is very useful for decision support, for example, the management of C. rosae with insecticidal sprays and the management of all of these pests using crop covers. There are other approaches, particularly those that need to be applied at sowing or transplanting, where knowledge of pest phenology and abundance is less necessary. Going forward, it is likely that the number of insecticidal control options available to European vegetable growers will diminish and they will need to move from a strategy which often involves using a single 'silver bullet' to a combination of approaches/tools with partial effects (applied within an IPM framework). For the less-effective, combined methods, accurate information about pest phenology and abundance and reliable decision support are likely to be extremely important.


The Stroop congruency effect is more observable under a speed strategy than an accuracy strategy.
Naming the ink color of an incongruent color word (e.g., RED printed in green) usually takes longer than naming the ink color of a color bar. However, when the ink matches the word (e.g., RED printed in red), naming tends to be faster. These phenomena are known as the STroop interference effect and the Stroop congruency effect, respectively. Although the interference effect has been robust and reliable across studies, the congruency effect tends to be elusive. It was hypothesized that this variation in outcomes might be related to subjects' response strategy. The experiment conducted to test this hypothesis induced either a speed or an accuracy strategy in two separate groups of subjects. Significant interference effects were found for both groups and the magnitudes did not differ. At the same time, the congruency effect was observed in the speed group but not in the accuracy group. These results suggest that researchers who wish to observe and study the Stroop congruency and interference effects should place special emphasis on speed. Implications of the study for a model of the Stroop effect are also discussed.


Trends in media use.
American youth are awash in media. They have television sets in their bedrooms, personal computers in their family rooms, and digital music players and cell phones in their backpacks. They spend more time with media than any single activity other than sleeping, with the average American eight- to eighteen-year-old reporting more than six hours of daily media use. The growing phenomenon of "media multitasking"--using several media concurrently--multiplies that figure to eight and a half hours of media exposure daily. Donald Roberts and Ulla Foehr examine how both media use and media exposure vary with demographic factors such as age, race and ethnicity, and household socioeconomic status, and with psychosocial variables such as academic performance and personal adjustment. They note that media exposure begins early, increases until children begin school, drops off briefly, then climbs again to peak at almost eight hours daily among eleven- and twelve-year-olds. Television and video exposure is particularly high among African American youth. Media exposure is negatively related to indicators of socioeconomic status, but that relationship may be diminishing. Media exposure is positively related to risk-taking behaviors and is negatively related to personal adjustment and school performance. Roberts and Foehr also review evidence pointing to the existence of a digital divide--variations in access to personal computers and allied technologies by socioeconomic status and by race and ethnicity. The authors also examine how the recent emergence of digital media such as personal computers, video game consoles, and portable music players, as well as the media multitasking phenomenon they facilitate, has increased young people's exposure to media messages while leaving media use time largely unchanged. Newer media, they point out, are not displacing older media but are being used in concert with them. The authors note which young people are more or less likely to use several media concurrently and which media are more or less likely to be paired with various other media. They argue that one implication of such media multitasking is the need to reconceptualize "media exposure."


Immunohistochemical study of N-epsilon-carboxymethyl lysine (CML) in human brain: relation to vascular dementia.
Advanced glycation end-products (AGEs) and their receptor (RAGE) occur in dementia of the Alzheimer's type and diabetic microvascular disease. Accumulation of AGEs relates to risk factors for vascular dementia with ageing, including hypertension and diabetes. Cognitive dysfunction in vascular dementia may relate to microvascular disease resembling that in diabetes. We tested if, among people with cerebrovascular disease, (1) those with dementia have higher levels of neuronal and vascular AGEs and (2) if cognitive dysfunction depends on neuronal and/or vascular AGE levels. Brain Sections from 25 cases of the OPTIMA (Oxford Project to Investigate Memory and Ageing) cohort, with varying degrees of cerebrovascular pathology and cognitive dysfunction (but only minimal Alzheimer type pathology) were immunostained for Nepsilon-(carboxymethyl)-lysine (CML), the most abundant AGE. The level of staining in vessels and neurons in the cortex, white matter and basal ganglia was compared to neuropsychological and other clinical measures. The probability of cortical neurons staining positive for CML was higher in cases with worse cognition (p = 0.01) or a history of hypertension (p = 0.028). Additionally, vascular CML staining related to cognitive impairment (p = 0.02) and a history of diabetes (p = 0.007). Neuronal CML staining in the basal ganglia related to a history of hypertension (p = 0.002). CML staining in cortical neurons and cerebral vessels is related to the severity of cognitive impairment in people with cerebrovascular disease and only minimal Alzheimer pathology. These findings support the possibility that cerebral accumulation of AGEs may contribute to dementia in people with cerebrovascular disease.


Comparing the responsiveness of functional outcome assessment measures for trauma registries.
Measuring long-term disability and functional outcomes after major trauma is not standardized across trauma registries. An ideal measure would be responsive to change but not have significant ceiling effects. The aim of this study was to compare the responsiveness of the Glasgow Outcome Scale (GOS), GOS-Extended (GOSE), Functional Independence Measure (FIM), and modified FIM in major trauma patients, with and without significant head injuries. Patients admitted to two adult Level I trauma centers in Victoria, Australia, who survived to discharge from hospital, were aged 15 years to 80 years with a blunt mechanism of injury, and had an estimated Injury Severity Score >15 on admission, were recruited for this prospective study. The instruments were administered at baseline (hospital discharge) and by telephone interview 6 months after injury. Measures of responsiveness, including effect sizes, were calculated. Bootstrapping techniques, and floor and ceiling effects, were used to compare the measures. Two hundred forty-three patients participated, of which 234 patients (96%) completed the study. The GOSE and GOS were the most responsive instruments in this major trauma population with effect sizes of 5.3 and 4.4, respectively. The GOSE had the lowest ceiling effect (17%). The GOSE was the instrument with greatest responsiveness and the lowest ceiling effect in a major trauma population with and without significant head injuries and is recommended for use by trauma registries for monitoring functional outcomes and benchmarking care. The results of this study do not support the use of the modified FIM for this purpose.


Patient-Reported Outcome Measures for Symptom Perception During a Cystic Fibrosis Exacerbation.
Symptom burden increases during pulmonary exacerbations of cystic fibrosis (CF), and patient-reported outcome measures (PROMs) are often used to evaluate symptoms as either primary or secondary outcomes. However, there is currently no guidance on the use of PROMs to assess symptom burden during pulmonary exacerbations. A systematic literature search was conducted to identify PROMs measuring symptom experience, management, or influencing factors, which were developed for CF patients and had been used at least once during pulmonary exacerbations. The PROMs included were assessed for relevance and psychometrics, according to the criteria of the United States FDA guideline and the Consensus-Based Standards for the Selection of Health Measurement Instruments (COSMIN) checklist. Five PROMs were identified, all measuring symptom perception. The CF Respiratory Symptom Diary (CFRSD) and the Symptom Scoring System were developed to assess symptom severity during pulmonary exacerbations. Of the other 3, which also included symptom scores of 2 quality of life measures, one assessed symptom severity exclusively, and 2 measured symptom severity in addition to other dimensions (such as symptom distress). All 5 instruments measured respiratory symptoms. Other relevant symptoms, such as energy and emotions, were covered by 4 instruments; pain and gastrointestinal symptoms were covered by 2 measures. All of the instruments demonstrated good internal consistency and sensitivity to change over a period up to 4 weeks. The symptom scores of the 2 quality of life measures with longer recall periods are not suitable for measuring assessed changes in a period of < 2 weeks. Criterion validity for gastrointestinal subscores has not been established. Discriminant validity was established in all of the instruments reviewed except for the Symptom Score System. Of the current PROMs used during CF pulmonary exacerbations, only 2 have been developed for this purpose, and only the CFRSD fulfilled all FDA guideline criteria. To date, there is no instrument that assesses exacerbation-specific symptom distress.


Phylogenetic analysis of a gene cluster encoding an additional, rhizobial-like type III secretion system that is narrowly distributed among Pseudomonas syringae strains.
The central role of Type III secretion systems (T3SS) in bacteria-plant interactions is well established, yet unexpected findings are being uncovered through bacterial genome sequencing. Some Pseudomonas syringae strains possess an uncharacterized cluster of genes encoding putative components of a second T3SS (T3SS-2) in addition to the well characterized Hrc1 T3SS which is associated with disease lesions in host plants and with the triggering of hypersensitive response in non-host plants. The aim of this study is to perform an in silico analysis of T3SS-2, and to compare it with other known T3SSs. Based on phylogenetic analysis and gene organization comparisons, the T3SS-2 cluster of the P. syringae pv. phaseolicola strain is grouped with a second T3SS found in the pNGR234b plasmid of Rhizobium sp. These additional T3SS gene clusters define a subgroup within the Rhizobium T3SS family. Although, T3SS-2 is not distributed as widely as the Hrc1 T3SS in P. syringae strains, it was found to be constitutively expressed in P. syringae pv phaseolicola through RT-PCR experiments. The relatedness of the P. syringae T3SS-2 to a second T3SS from the pNGR234b plasmid of Rhizobium sp., member of subgroup II of the rhizobial T3SS family, indicates common ancestry and/or possible horizontal transfer events between these species. Functional analysis and genome sequencing of more rhizobia and P. syringae pathovars may shed light into why these bacteria maintain a second T3SS gene cluster in their genome.


Fa1p is a 171 kDa protein essential for axonemal microtubule severing in Chlamydomonas.
A key event in deflagellation or deciliation is the severing of the nine outer-doublet axonemal microtubules at a specific site in the flagellar transition zone. Previous genetic analysis revealed three genes that are essential for deflagellation in Chlamydomonas. We have now identified the first of these products, Fa1p, a protein required for Ca(2+)-dependent, axonemal microtubule severing. Genetic mapping and the availability of a tagged allele allowed us to physically map the gene to the centromere-proximal domain of the mating-type locus. We identified clones of Chlamydomonas genomic DNA that rescued the Ca(2+)-dependent axonemal microtubule severing defect of fa1 mutants. The FA1 cDNA, obtained by RT-PCR, encodes a novel protein of 171 kDa, which is predicted to contain an amino-terminal coiled-coil domain and three Ca(2+)/calmodulin binding domains. By western analysis and subcellular fractionation, the FA1 product is enriched in flagellar-basal body complexes. Based on these observations and previous studies, we hypothesize that a Ca(2+)-activated, Ca(2+)-binding protein binds Fa1p leading ultimately to the activation of axonemal microtubule severing.


Undetectable Tumor Cell-Free DNA in a Patient With Metastatic Breast Cancer With Complete Response and Long-Term Remission.
The ability to serially monitor tumor-derived cell-free DNA (cfDNA) brings with it the potential to measure response to anticancer therapies and detect minimal residual disease (MRD). This report describes a patient with HER2-positive metastatic breast cancer with an exceptional response to trastuzumab and nab-paclitaxel who remains in complete remission several years after cessation of therapy. Next-generation sequencing of the patient's primary tumor tissue showed several mutations, including an oncogenic hotspot PIK3CA mutation. A sample of cfDNA was collected 6 years after her last therapy and then analyzed for mutant PIK3CA using digital PCR. No detectable mutations associated with the primary tumor were found despite assaying >10,000 genome equivalents, suggesting that the patient had achieved a molecular remission. Results of this case study suggest that serial monitoring of MRD using liquid biopsies could provide a useful method for individualizing treatment plans for patients with metastatic disease with extreme responses to therapy. However, large-scale clinical studies are needed to validate and implement these techniques for patient care.


A 10-year review of child injury hospitalisations, health outcomes and treatment costs in Australia.
Childhood injury is a leading cause of hospitalisation, yet there has been no comprehensive examination of child injury and survival over time in Australia. To examine the characteristics, temporal trend and survival for children who were hospitalised as a result of injury in Australia. A retrospective examination of linked hospitalisation and mortality data for injured children aged 16 years or less during 1 July 2001 to 30 June 2012. Negative binomial regression examined change in injury hospitalisation trends. Cox proportional hazard regression examined the association of risk factors on 30-day survival. There were 6 86 409 injury hospitalisations, with an age-standardised rate of 1489 per 1 00 000 population (95% CI 1485.3 to 1492.4) in Australia. Child injury hospitalisation rates did not change over the 10-year period. For every severely injured child, there are at least 13 children hospitalised with minor or moderate injuries. The total cost of child injury hospitalisations was $A2.1 billion (annually $A212 million). Falls (38.4%) were the most common injury mechanism. Factors associated with a higher risk of 30-day mortality were: child was aged ≤10 years, higher injury severity, head injury, injured in a transport incident or following drowning and submersion or other threats to breathing, during self-harm and usual residence was regional/remote Australia. Childhood injury hospitalisation rates have not reduced in 10 years. Children's patterns of injury change with age, and priorities for injury prevention alter according to developmental stages. The development of a national multisectorial childhood injury monitoring and prevention strategy in Australia is long overdue.


Cellular stress response: a novel target for chemoprevention and nutritional neuroprotection in aging, neurodegenerative disorders and longevity.
The predominant molecular symptom of aging is the accumulation of altered gene products. Moreover, several conditions including protein, lipid or glucose oxidation disrupt redox homeostasis and lead to accumulation of unfolded or misfolded proteins in the aging brain. Alzheimer's and Parkinson's diseases or Friedreich ataxia are neurological diseases sharing, as a common denominator, production of abnormal proteins, mitochondrial dysfunction and oxidative stress, which contribute to the pathogenesis of these so called "protein conformational diseases". The central nervous system has evolved the conserved mechanism of unfolded protein response to cope with the accumulation of misfolded proteins. As one of the main intracellular redox systems involved in neuroprotection, the vitagene system is emerging as a neurohormetic potential target for novel cytoprotective interventions. Vitagenes encode for cytoprotective heat shock proteins (Hsp) Hsp70 and heme oxygenase-1, as well as thioredoxin reductase and sirtuins. Nutritional studies show that ageing in animals can be significantly influenced by dietary restriction. Thus, the impact of dietary factors on health and longevity is an increasingly appreciated area of research. Reducing energy intake by controlled caloric restriction or intermittent fasting increases lifespan and protects various tissues against disease. Genetics has revealed that ageing may be controlled by changes in intracellular NAD/NADH ratio regulating sirtuin, a group of proteins linked to aging, metabolism and stress tolerance in several organisms. Recent findings suggest that several phytochemicals exhibit biphasic dose responses on cells with low doses activating signaling pathways that result in increased expression of vitagenes encoding survival proteins, as in the case of the Keap1/Nrf2/ARE pathway activated by curcumin and NAD/NADH-sirtuin-1 activated by resveratrol. Consistently, the neuroprotective roles of dietary antioxidants including curcumin, acetyl-L-carnitine and carnosine have been demonstrated through the activation of these redox-sensitive intracellular pathways. Although the notion that stress proteins are neuroprotective is broadly accepted, still much work needs to be done in order to associate neuroprotection with specific pattern of stress responses. In this review the importance of vitagenes in the cellular stress response and the potential use of dietary antioxidants in the prevention and treatment of neurodegenerative disorders is discussed.


A maximum likelihood approach to continuous speech recognition.
Speech recognition is formulated as a problem of maximum likelihood decoding. This formulation requires statistical models of the speech production process. In this paper, we describe a number of statistical models for use in speech recognition. We give special attention to determining the parameters for such models from sparse data. We also describe two decoding methods, one appropriate for constrained artificial languages and one appropriate for more realistic decoding tasks. To illustrate the usefulness of the methods described, we review a number of decoding results that have been obtained with them.


BLAST: a more efficient report with usability improvements.
The Basic Local Alignment Search Tool (BLAST) website at the National Center for Biotechnology (NCBI) is an important resource for searching and aligning sequences. A new BLAST report allows faster loading of alignments, adds navigation aids, allows easy downloading of subject sequences and reports and has improved usability. Here, we describe these improvements to the BLAST report, discuss design decisions, describe other improvements to the search page and database documentation and outline plans for future development. The NCBI BLAST URL is http://blast.ncbi.nlm.nih.gov.


A comparison of nonoperative vs. Endobutton repair of distal biceps ruptures.
The aim of this study was to compare the outcome of patients who have undergone distal biceps tendon repair by a single-incision Endobutton fixation technique with the results of another cohort of patients who elected not to undergo surgery for distal biceps tendon rupture. A retrospective cohort study was performed of patients diagnosed with distal biceps ruptures, repaired with an Endobutton (Smith & Nephew, Andover, MA, USA) technique or treated nonoperatively by the senior surgeon (S.A.S.). With a minimum follow-up of 6 months, a routine elbow examination, radiographs, and functional questionnaires were performed. Isometric supination, flexion, and grip strength was measured using a BTE machine (Baltimore Therapeutic Equipment, Hanover, MD, USA). There were 47 patients available for follow-up with 50 distal biceps ruptures; 40 ruptures have undergone repair, and 10 have been managed nonoperatively. Three patients had sustained bilateral ruptures. There was a significant difference in flexion and supination isometric strength between the operative and nonoperative cohorts compared with the uninjured contralateral side (92.94% vs. 70.65%, P = .01512; 87.91% vs 59.11%, P = .00414, respectively). The difference in grip strengths between the 2 cohorts compared with the uninjured side was not significant (100.00% vs. 79.16%; P = .16002). The operated cohort had significantly better QuickDASH score, Oxford Elbow Score, and Mayo Elbow Performance Score (6.29 vs. 14.10, P = .02123; 44.71 vs. 38.70, P = .00429; 93.13 vs. 84.50, P = .01423). Repair of distal biceps ruptures using an Endobutton fixation results in nearly normal return of strength and function, which is significantly better than in those managed nonoperatively. Level III, Retrospective Cohort Design; Treatment Study.


In vivo Evaluation of Two Thiazolidin-4-one Derivatives in High Sucrose Diet Fed Pre-diabetic Mice and Their Modulatory Effect on AMPK, Akt and p38 MAP Kinase in L6 Cells.
We had previously demonstrated the anti-diabetic potential and pancreatic protection of two thiazolidin-4-one derivatives containing nicotinamide moiety (NAT-1 and NAT-2) in STZ-induced diabetic mice. However, due to the limitations of the STZ model, we decided to undertake a detailed evaluation of anti-diabetic potential of the molecules on a high sucrose diet (HSD) fed diabetic mouse model. Further, in vitro mechanistic studies on the phosphorylation of AMPK, Akt and p38 MAP kinase in L6 myotubes and anti-inflammatory studies in RAW264.7 mouse monocyte macrophage cells were performed. 15 months of HSD induced fasting hyperglycaemia and impaired glucose tolerance in mice. Treatment with NAT-1 and NAT-2 (100 mg/kg) for 45 days significantly improved the glucose tolerance and lowered fasting blood glucose levels compared to untreated control. An improvement in the elevated triglycerides and total cholesterol levels, and favorable rise in HDL cholesterol were also observed with test drug treatment. Also, no major changes were observed in the liver (albumin, AST and ALT) and kidney (creatinine and urea) parameters. This was further confirmed in their respective histology profiles which revealed no gross morphological changes. In L6 cells, significant phosphorylation of Akt and p38 MAP kinase proteins were observed with 100 μM of NAT-1 and NAT-2 with no significant changes in phosphorylation of AMPK. The molecules failed to exhibit anti-inflammatory activity as observed by their effect on the generation of ROS and nitrite, and nuclear levels of NF-κB in LPS-stimulated RAW264.7 cells. In summary, the molecules activated Akt and p38 MAP kinase which could have partly contributed to their anti-hyperglycaemic and hypolipidemic activities in vivo.


Cessation of attention deficit hyperactivity disorder drugs in the young (CADDY)--a pharmacoepidemiological and qualitative study.
To estimate the prevalence of attention deficit hyperactivity disorder (ADHD) pharmacological treatment, and its demographic and clinical details, and to estimate the proportion of patients in the target group who stopped ADHD treatment and investigate possible factors for continuation or cessation of treatment. A pharmacoepidemiological study using an automated database and a qualititative study using patient interviews. Part 1 was a pharmacoepidemiological study that provided accurate data on use and cessation of ADHD drugs. Part 2 was an in-depth interview study to investigate the reasons, processes and outcomes of treatment cessation. Part 1: primary care using the General Practice Research Database (GPRD). Part 2: secondary and tertiary care paediatric clinics, child and adolescent mental health and adult mental health clinics in London, Nottingham, Dundee and Liverpool. Part 1: patients were 15-21 years old during the study period (1 January 2001 and 31 December 2004), had at least one prescription for methylphenidate, dexamfetamine or atomoxetine and had at least 1 year of research-standard data available in the GPRD. Part 2: patients fulfilled Part 1 criteria, had a diagnosis of ADHD as detected by a predefined algorithm and had been treated with methylphenidate, dexamfetamine or atomoxetine for at least 1 year. Child and adolescent psychiatrists, adult psychiatrists and paediatricians involved in the treatment of young people with ADHD were also interviewed as part of the study. Part 1: prevalence of prescribing averaged across all ages increased eightfold, from 0.26 per 1000 patients in 1999 to 2.07 per 1000 patients in 2006. The increase in prevalence in the younger patients was less evident in the older patients. Prevalence in 15-year-old males receiving a study drug prescription increased from 1.32 per 1000 patients in 1999 to 8.31 per 1000 patients in 2006, whereas the prevalence in 21-year-olds rose from 0 per 1000 patients in 1999 to 0.43 per 1000 patients in 2006. Survival analysis showed that the rate of treatment cessation largely exceeded the estimated rate of persistence of ADHD. The reduction in prescribing was most noticeable between 16 and 17 years of age. Kaplan-Meier analysis showed that approximately 18% of patients restarted treatment if they had stopped treatment after the age of 15. Patients who restarted treatment were more likely to restart within the first year following treatment cessation. Part 2: the Child Health and Illness Profile (CHIP) was chosen as the quality of life questionnaire for the Part 2 study because the CHIP-CE scale has been validated in children with ADHD in the UK. Because of the age range of participants, the adolescent version (CHIP-AE) was administered to patients after interview. Of the 15, a total of nine patients finished the questionnaire. Interviews showed that although some young people felt able to cope after stopping medication, others felt the need to restart to control symptoms. Some patients had difficulty re-engaging with services and clinicians recognised the lack of services for young adults. Patients continuing on treatment considered cessation as an option for the future, but were concerned about the process of stopping and its impact on behaviour. Part 1 study demonstrated that the prevalence of prescribing by GPs to patients with ADHD dropped significantly from age 15 to 21. The fall in prescribing was greater than the reported age-related decrease in symptoms, raising the possibility that treatment is prematurely discontinued in some young adults where ADHD symptoms persist. Part 2 of the study identified that some young adults had difficulty in obtaining treatment after discharge from paediatric services. Future work should include randomised placebo-controlled trials into long-term treatment with stimulants, particularly methylphenidate.


Association of the Endobiont Double-Stranded RNA Virus LRV1 With Treatment Failure for Human Leishmaniasis Caused by Leishmania braziliensis in Peru and Bolivia.
Cutaneous and mucosal leishmaniasis, caused in South America by Leishmania braziliensis, is difficult to cure by chemotherapy (primarily pentavalent antimonials [Sb(V)]). Treatment failure does not correlate well with resistance in vitro, and the factors responsible for treatment failure in patients are not well understood. Many isolates of L. braziliensis (>25%) contain a double-stranded RNA virus named Leishmaniavirus 1 (LRV1), which has also been reported in Leishmania guyanensis, for which an association with increased pathology, metastasis, and parasite replication was found in murine models. Here we probed the relationship of LRV1 to drug treatment success and disease in 97 L. braziliensis-infected patients from Peru and Bolivia. In vitro cultures were established, parasites were typed as L. braziliensis, and the presence of LRV1 was determined by reverse transcription-polymerase chain reaction, followed by sequence analysis. LRV1 was associated significantly with an increased risk of treatment failure (odds ratio, 3.99; P = .04). There was no significant association with intrinsic Sb(V) resistance among parasites, suggesting that treatment failure arises from LRV1-mediated effects on host metabolism and/or parasite survival. The association of LRV1 with clinical drug treatment failure could serve to guide more-effective treatment of tegumentary disease caused by L. braziliensis.


Characterization of a Novel Insecticidal Protein Cry9Cb1 from Bacillus thuringiensis.
In recent decades, there have been increasing reports of insect resistance in Bacillus thuringiensis (Bt) crops. Alternative use of Cry toxins, with high insecticidal activity and different mechanisms of action, may be an important strategy to manage this resistance. Cry9 protein, with high toxicity to the lepidopteran pests and no cross-resistance with commercial Cry1 proteins, is a valuable relevant resource. A novel insecticidal protein, MP1489, subsequently named as Cry9Cb1, with 88% amino acid sequence identity with Cry9Ca1, was identified from Bt strain SP663; it exhibited high insecticidal activity against Plutella xylostella, Ostrinia furnacalis, and Chilo suppressalis and no cross-resistance with Cry1Fa in Ostrinia furnacalis. Its minimal active fragments against Plutella xylostella and Ostrinia furnacalis were identified to be 72T-657V and 68D-655A, respectively; food-safety assessment showed no sequence homology with any known allergen and rapid degradation and inactivation by both heat and the gastrointestinal environment. Therefore, Cry9Cb1 is proposed to have a brilliant prospect as an insecticidal protein in agriculture.


3T proton MR Spectroscopy evaluation of spinal cord lesions.
The objective of this study was to evaluate intramedullary spinal cord lesions using magnetic resonance spectroscopy and correlate the results with histo-pathological examination (HPE). Approval for this study was obtained from our institute ethical committee. Overall, 50 patients were recruited (29 male and 21 female), with a maximum age of 53 years and minimum age of 7 years. The mean age group of the study was 33 years. Standard magnetic resonance imaging (MRI) spine was done on a Siemens Skyra 3Tesla MRI scanner. MR Spectroscopy (MRS) was performed for all patients with intramedullary spinal lesions after getting written consent. It was performed using single-voxel method. The change in the metabolite peak was observed in each case and the results were compared with HPE. These collected data were analyzed using SPSS 16.0 version. Descriptive statistics, frequency analysis, and percentage analysis were used for categorical variables; and for continuous variables, mean and standard deviation were analyzed. McNemar's test was used to find the significance between conventional MRI MRS. In the above statistical tool, the probability value 0.05 is considered as significant level. From our study, we observed that by applying routine MRI sequences alone, we could only detect around 58% of the cases correctly. However, when MRS was done along with the conventional MR imaging, the number of cases detected significantly increased to 84%. By applying McNemar's test and comparing the conventional MRI and MRS with HPE, it was found that statistically significant difference exists with P value of 0.007. MRS of the spinal cord is a promising tool for research and diagnosis because it can provide additional information complementary to other non-invasive imaging methods. It is an emerging tool and adds new biomarker information for characterization of spinal cord tumors, to differentiate benign from malignant lesions and to prevent unnecessary biopsies and surgeries.


An epidemiological study of traumatic brachial plexus injury patients treated at an Indian centre.
Epidemiological studies on traumatic brachial plexus injuries are few and these studies help us to improve the treatment, rehabilitation of these patients and to allocate the resources required in their management. Epidemiological factors can vary in different countries. We wanted to know the situation in an Indian centre. Data regarding age, sex, affected side, mode of injury, distribution of paralysis, associated injuries, pain at the time of presentation and the index procedure they underwent were collected from 304 patients. Additional data like the vehicle associated during the accident, speed of the vehicle during the accident, employment status and integration into the family were collected in 144 patients out of the 304 patients. Road traffic accidents accounted for 94% of patients and of the road traffic accidents 90% involved two wheelers. Brachial plexus injury formed a part of multitrauma in 54% of this study group and 46% had isolated brachial plexus injury. Associated injuries like fractures, vascular injuries and head injuries are much less probably due to the lower velocity of the vehicles compared to the western world. The average time interval from the date of injury to exploration of the brachial plexus was 127 days and 124 (40.78%) patients presented to us within this duration. Fifty-seven per cent had joined back to work by an average of 8.6 months. It took an average of 6.8 months for the global brachial plexus-injured patients to write in their non-dominant hand.


Serine proteases in rodent hippocampus.
Brain serine proteases are implicated in developmental processes, synaptic plasticity, and in disorders including Alzheimer's disease. The spectrum of the major enzymes expressed in brain has not been established previously. We now present a systematic study of the serine proteases expressed in adult rat and mouse hippocampus. Using a combination of techniques including polymerase chain reaction amplification and Northern blotting we show that tissue-type plasminogen activator (t-PA) is the major species represented. Unexpectedly, the next most abundant species were RNK-Met-1, a lymphocyte protease not reported previously in brain, and two new family members, BSP1 (brain serine protease 1) and BSP2. We report full-length sequences of the two new proteases; homologies indicate that these are of tryptic specificity. Although BSP2 is expressed in several brain regions, BSP1 expression is strikingly restricted to hippocampus. Other enzymes represented, but at lower levels, included elastase IV, proteinase 3, complement C2, chymotrypsin B, chymotrypsin-like protein, and Hageman factor. Although thrombin and urokinase-type plasminogen activator were not detected in the primary screen, low level expression was confirmed using specific polymerase chain reaction primers. In contrast, and despite robust expression of t-PA, the usual t-PA substrate plasminogen was not expressed at detectable levels.


Dysregulation of follicle development in a mouse model of premature ovarian insufficiency.
Premature ovarian insufficiency (POI) occurs in 1% of reproductive-age women. The ovarian manifestation ranges from the presence of a variable population of follicles (follicular) to the absence of follicles (afollicular), and in the majority of cases the cause is unknown. A transgenic mouse model of follicular POI, the Double Mutant (DM), arises from oocyte-specific deletion of Mgat1 and C1galt1 required for the generation of O- and N-glycans. DM females are subfertile at 6 weeks, infertile by 9 weeks and exhibit POI by 12 weeks of age. In this study we investigate the cause of the reduced fertility at 6 weeks and infertility at 9 weeks of DM females. Ovary sections were used to analyse follicle and corpora lutea (CL) numbers, apoptosis, and levels of laminin and 3β-hydroxysteroid dehydrogenase using immunohistochemistry. After POI, DM females unexpectedly remained sexually receptive. At both 6 and 9 weeks, DM ovaries contained more primary follicles, however, at 9 weeks DM follicles were proportionally healthier, revealed by TUNEL analysis compared with Controls. In 9 week DM ovaries (collected post-mating), secondary follicles had theca and basal lamina structure abnormalities, whilst preovulatory follicles failed to ovulate resulting in the presence of numerous luteinised unruptured follicles, indicative of ovulation failure. Finally, DM ovaries contained more regressing CL with decreased luteal cell apoptosis indicative of a defect in CL regression. Identifying these follicular modifications have provided insight into the aetiology of a model of POI and highlight targets to investigate with the hope of developing new fertility treatments.


A single fixation protocol for proteome-wide immunofluorescence localization studies.
Immunofluorescence microscopy is a valuable tool for analyzing protein expression and localization at a subcellular level thus providing information regarding protein function, interaction partners and its role in cellular processes. When performing sample fixation, parameters such as difference in accessibility of proteins present in various cellular compartments as well as the chemical composition of the protein to be studied, needs to be taken into account. However, in systematic and proteome-wide efforts, a need exists for standard fixation protocol(s) that works well for the majority of all proteins independent of subcellular localization. Here, we report on a study with the goal to find a standardized protocol based on the analysis of 18 human proteins localized in 11 different organelles and subcellular structures. Six fixation protocols were tested based on either dehydration by alcohols (methanol, ethanol or iso-propanol) or cross-linking by paraformaldehyde followed by detergent permeabilization (Triton X-100 or saponin) in three human cell lines. Our results show that cross-linking is essential for proteome-wide localization studies and that cross-linking using paraformaldehyde followed by Triton X-100 permeabilization successfully can be used as a single fixation protocol for systematic studies.


A common substitution (Asn291Ser) in lipoprotein lipase is associated with increased risk of ischemic heart disease.
Lipoprotein lipase degrades triglycerides in plasma and as a byproduct produces HDL particles. Genetic variation in lipoprotein lipase may therefore affect cardiovascular risk. We tested 9,214 men and women from a general population sample and 948 patients with ischemic heart disease for the Asn291Ser substitution in lipoprotein lipase. The allele frequency in the general population was 0.024 and 0.026 for women and men, respectively. In comparison with noncarriers, female heterozygous probands had increased plasma triglycerides (delta = 0.23 mmol/liter), while HDL cholesterol was reduced in both female and male carriers (delta = 0.18 mmol/liter and delta = 0.11 mmol/liter, respectively). A similar phenotype was found in six homozygous carriers. On multiple logistic regression analysis, plasma triglycerides and HDL cholesterol were independent predictors of ischemic heart disease in both genders. On univariate analysis, odds ratios for ischemic heart disease in probands were 1.89 in women (95% CI: 1.19-3.01) and 0.90 in men (95% CI: 0.62-1.31), and on multivariate analysis were 1.98 in women (95% CI: 1.11-3.53) and 1.02 in men (95% CI: 0.65-1.60). This study demonstrates that a single common mutation in the lipoprotein lipase gene is associated with elevated plasma triglycerides and reduced HDL cholesterol levels, whereby carriers, in particular women, seem to be predisposed to ischemic heart disease. It cannot be excluded, however, that male carriers of this substitution may represent a subset of low-HDL individuals without raised triglycerides not predisposed to ischemic heart disease.


Protocol for the Data-Linkage Alcohol Cohort Study (DACS): investigating mortality, morbidity and offending among people with an alcohol-related problem using linked administrative data.
The aims of this program of research are to use linked health and law enforcement data to describe individuals presenting to emergency and inpatient healthcare services with an acute alcohol harm or problematic alcohol use; measure their health service utilisation and law enforcement engagement; and quantify morbidity, mortality, offending and incarceration. We will assemble a retrospective cohort of people presenting to emergency departments and/or admitted to hospitals between 1 January 2005 and 31 December 2014 in New South Wales, Australia with a diagnosis denoting an acute alcohol harm or problematic alcohol use. We will link these data with records from other healthcare services (eg, community-based mental healthcare data, cancer registry), mortality, offending and incarceration data sets. The four overarching areas for analysis comprise: (1) describing the characteristics of the cohort at their first point of contact with emergency and inpatient hospital services in the study period with a diagnosis indicating an acute alcohol harm and/or problematic alcohol use; (2) quantifying health service utilisation and law enforcement engagement; (3) quantifying rates of mortality, morbidity, offending and incarceration; and (4) assessing predictors (eg, age, sex) of mortality, morbidity, offending and incarceration among this cohort. Ethics approval has been provided by the New South Wales Population and Health Services Research Ethics Committee. We will report our findings in accordance with the REporting of studies Conducted using Observational Routinely collected health Data (RECORD) statement and Guidelines for Accurate and Transparent Health Estimates Reporting (GATHER) where appropriate. We will publish data in tabular, aggregate forms only. We will not disclose individual results. We will disseminate project findings at scientific conferences and in peer-reviewed journals. We will aim to present findings to relevant stakeholders (eg, addiction medicine and emergency medicine specialists, policy makers) to maximise translational impact of research findings.


Large scale identification and categorization of protein sequences using structured logistic regression.
Structured Logistic Regression (SLR) is a newly developed machine learning tool first proposed in the context of text categorization. Current availability of extensive protein sequence databases calls for an automated method to reliably classify sequences and SLR seems well-suited for this task. The classification of P-type ATPases, a large family of ATP-driven membrane pumps transporting essential cations, was selected as a test-case that would generate important biological information as well as provide a proof-of-concept for the application of SLR to a large scale bioinformatics problem. Using SLR, we have built classifiers to identify and automatically categorize P-type ATPases into one of 11 pre-defined classes. The SLR-classifiers are compared to a Hidden Markov Model approach and shown to be highly accurate and scalable. Representing the bulk of currently known sequences, we analysed 9.3 million sequences in the UniProtKB and attempted to classify a large number of P-type ATPases. To examine the distribution of pumps on organisms, we also applied SLR to 1,123 complete genomes from the Entrez genome database. Finally, we analysed the predicted membrane topology of the identified P-type ATPases. Using the SLR-based classification tool we are able to run a large scale study of P-type ATPases. This study provides proof-of-concept for the application of SLR to a bioinformatics problem and the analysis of P-type ATPases pinpoints new and interesting targets for further biochemical characterization and structural analysis.


Cryptogenic Cirrhosis and Sitosterolemia: A Treatable Disease If Identified but Fatal If Missed.
Sitosterolemia is an autosomal recessive metabolic disease caused by mutations in ABCG5 or ABCG8 genes which encode for the (ATP)-binding cassette (ABC) transporters that are responsible for the trafficking of xenosterols. Liver involvement is not a recognized manifestation of this disease, and cirrhosis has been reported only once in the medical literature. We describe a fatal case of a 21-year old South Asian male who presented with decompensated cirrhosis, and biochemical abnormalities consistent with sitosterolemia. Genetic testing showed a homozygous pathogenic mutation in ABCG5, confirming the diagnosis. Sitosterolemia is a rare, but likely under-recognized condition, and a high degree of suspicion is imperative to make the diagnosis. We propose that sitosterolemia should be included in the differential diagnosis for patients with cryptogenic cirrhosis, especially as there are effective oral therapies to treat this condition. Newly diagnosed sitosterolemia patients should undergo a thorough hepatology evaluation and follow-up to evaluate for the presence, development, and progression of any hepatic involvement.


Mobilization of iron from ferritin: new steps and details.
Much evidence indicates that iron stored in ferritin is mobilized through protein degradation in lysosomes, but concerns about this process have lingered, and the mechanistic details of its aspects are lacking. In the studies presented here, 59Fe-labeled ferritin was induced by preloading hepatic (HepG2) cells with radiolabeled Fe. Placing these cells in a medium containing desferrioxamine resulted in the loss of ferritin-59Fe, but adding high concentrations of reducing agents or modulating the internal GSH concentration failed to alter the rates of ferritin-59Fe release. Confocal microscopy showed that Fe deprivation increased the movement of ferritin into lysosomes and hyperaccumulation was observed when lysosomal proteolysis was inhibited. It also resulted in the rapid movement of DMT1 to lysosomes, which was inhibited by bafilomycin. Ferrihydrite crystals isolated from purified rat liver/spleen ferritin were solubilized at pH 5 and 7 by GSH, ascorbate, citrate and lysosomal fluids obtained from livers and J774a.1 macrophages. The inhibition of DMT1/Nramp2 and siRNA knockdown of Nramp1 each reduced the transfer of 59Fe from lysosomes to the cytosol; and hepatocyte-specific knockout of DMT1 in mice prevented the release of Fe from the liver responding to EPO treatment, but did not inhibit lysosomal ferritin degradation. We conclude that ferritin-Fe mobilization does not occur through changes in cellular concentrations of reducing/chelating agents but by the coordinated movement of ferritin and DMT1 to lysosomes, where the ferrihydrite crystals exposed by ferritin degradation dissolve in the lysosomal fluid, and the reduced iron is transported back to the cytosol via DMT1 in hepatocytes, and by both DMT1 and Nramp1 in macrophages, prior to release into the blood or storage in ferritin.


Remote sensing reveals unprecedented sublethal impacts of a 40-year-old oil spill on mangroves.
Oil spills cause long-lasting mangrove loss, threatening their conservation and ecosystem services worldwide. Oil spills impact mangrove forests at various spatial and temporal scales. Yet, their long-term sublethal effects on trees remain poorly documented. Here, we explore these effects based on one of the largest oil spills ever recorded, the Baixada Santista pipeline leak, which hit the mangroves of the Brazilian southeastern coast in 1983. Historical, Landsat-derived normalized difference vegetation index (NDVI) maps over the spilled mangrove reveal a large dieback of trees within a year following the oil spill, followed by a eight-year recolonization period and a stabilization of the canopy cover, however 20-30% lower than initially observed. We explain this permanent loss by an unexpected persistence of oil pollution in the sediments based on visual and geochemical evidence. Using field spectroscopy and cutting-edge drone hyperspectral imaging, we demonstrate how the continuous exposure of mangrove trees to high levels of pollution affects their health and productivity in the long term, by imposing permanent stressful conditions. Our study also reveals that tree species differ in their sensitivity to oil, giving the most tolerant ones a competitive advantage to recolonize spilled mangroves. By leveraging drone laser scanning, we estimate the loss of forest biomass caused by the oil spill to be 9.8-91.2 t ha-1, corresponding to 4.3-40.1 t C ha-1. Based on our findings, we encourage environmental agencies and lawmakers to consider the sublethal effects of oil spills on mangroves in the environmental cost of these accidents. We also encourage petroleum companies to use drone remote sensing in monitoring routines and oil spill response planning to improve mangrove preservation and impact assessment.


STRFs in primary auditory cortex emerge from masking-based statistics of natural sounds.
We investigate how the neural processing in auditory cortex is shaped by the statistics of natural sounds. Hypothesising that auditory cortex (A1) represents the structural primitives out of which sounds are composed, we employ a statistical model to extract such components. The input to the model are cochleagrams which approximate the non-linear transformations a sound undergoes from the outer ear, through the cochlea to the auditory nerve. Cochleagram components do not superimpose linearly, but rather according to a rule which can be approximated using the max function. This is a consequence of the compression inherent in the cochleagram and the sparsity of natural sounds. Furthermore, cochleagrams do not have negative values. Cochleagrams are therefore not matched well by the assumptions of standard linear approaches such as sparse coding or ICA. We therefore consider a new encoding approach for natural sounds, which combines a model of early auditory processing with maximal causes analysis (MCA), a sparse coding model which captures both the non-linear combination rule and non-negativity of the data. An efficient truncated EM algorithm is used to fit the MCA model to cochleagram data. We characterize the generative fields (GFs) inferred by MCA with respect to in vivo neural responses in A1 by applying reverse correlation to estimate spectro-temporal receptive fields (STRFs) implied by the learned GFs. Despite the GFs being non-negative, the STRF estimates are found to contain both positive and negative subfields, where the negative subfields can be attributed to explaining away effects as captured by the applied inference method. A direct comparison with ferret A1 shows many similar forms, and the spectral and temporal modulation tuning of both ferret and model STRFs show similar ranges over the population. In summary, our model represents an alternative to linear approaches for biological auditory encoding while it captures salient data properties and links inhibitory subfields to explaining away effects.


Identification and mechanosensitivity of viscerofugal neurons.
Enteric viscerofugal neurons are interneurons with cell bodies in the gut wall; they project to prevertebral ganglia where they provide excitatory synaptic drive to sympathetic neurons which control intestinal motility and secretion. Here, we studied the mechanosensitivity and firing of single, identified viscerofugal neurons in guinea-pig distal colon. Flat sheet preparations of gut were set up in vitro and conventional extracellular recordings made from colonic nerve trunks. The nicotinic agonist, 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP) (1mM), was locally pressure ejected onto individual myenteric ganglia. In a few ganglia, DMPP promptly evoked firing in colonic nerves. Biotinamide filling of colonic nerves revealed that DMPP-responsive sites corresponded to viscerofugal nerve cell bodies. This provides a robust means to positively identify viscerofugal neuron firing. Of 15 single units identified in this way, none responded to locally-applied capsaicin (1 μM). Probing with von Frey hairs at DMPP-responsive sites reliably evoked firing in all identified viscerofugal neurons (18/18 units tested; 0.8-5 mN). Circumferential stretch of the preparation increased firing in all 14/14 units (1-5 g, p<0.05). Both stretch and von Frey hair responses persisted in Ca(2+)-free solution (6 mM Mg(2+), 1mM EDTA), indicating that viscerofugal neurons are directly mechanosensitive. To investigate their adequate stimulus, circular muscle tension and length were independently modulated (BAY K8644, 1 μM and 10 μM, respectively). Increases in intramural tension without changes in length did not affect firing. However, contraction-evoked shortening, under constant load, significantly decreased firing (p<0.001). In conclusion, viscerofugal neuron action potentials contribute to recordings from colonic nerve trunks, in vitro. They provide a significant primary afferent output from the colon, encoding circumferential length, largely independent of muscle tension. All viscerofugal neurons are directly mechanosensitive, although they have been reported to receive synaptic inputs. In short, viscerofugal neurons combine interneuronal function with length-sensitive mechanosensitivity.


Acceptability of Chairside Screening for Racial/Ethnic Minority Older Adults: A Qualitative Study.
An aging and more racially and ethnically diverse population, coupled with changes in the health care policy environment, is demanding that the dental profession both redirect and expand its focus. Challenges include providing comprehensive care for patients with complicated medical needs while improving access to care for underserved groups. The purpose of this study is to examine the acceptability of screening for hypertension and diabetes in the dental setting for African American, Puerto Rican, and Dominican older adults who attend senior centers in northern Manhattan, New York City. Focus groups were conducted with 194 racial/ethnic minority men and women aged 50 y and older living in northern Manhattan who participated in 1 of 24 focus group sessions about improving oral health for older adults. All groups were digitally audio-recorded and transcribed for analysis. Groups that were conducted in Spanish were transcribed first in Spanish and then translated into English. Analysis of the transcripts was conducted using thematic content analysis. Five themes were manifest in the data regarding the willingness of racial/ethnic minority older adults to receive hypertension and diabetes screening as part of routine dental visits: 1) chairside screening is acceptable, 2) screening is routine for older adults, 3) the interrelationship between oral and general health is appreciated, 4) chairside screening has perceived benefits, and 5) chairside screening may reduce dental anxiety. Reservations centered on 4 major themes: 1) dental fear may limit the acceptability of chairside screening, 2) there is a perceived lack of need for dental care and chairside screening, 3) screening is available elsewhere, and 4) mistrust of dental providers as primary care providers. This study provides novel evidence of the acceptability of screening for hypertension and diabetes in the dental setting among urban racial/ethnic minority senior center attendees. Knowledge Transfer Statement: The results of this study may be used by oral health providers when deciding whether to conduct chairside screening for medical conditions such as hypertension and diabetes that could affect, or be affected by, the oral health of their patients. Patient experiences of care-along with clinical outcomes, avoidable hospital admissions, equity of services, and costs-are important outcomes to consider in meeting the needs of an aging and racially and ethnically diverse US population.