A CXCR4-targeted nanocarrier achieves highly selective tumor uptake in diffuse large B-cell lymphoma mouse models.
One-third of diffuse large B-cell lymphoma patients are refractory to initial treatment or relapse after rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisone chemotherapy. In these patients, CXCR4 overexpression (CXCR4+) associates with lower overall and disease-free survival. Nanomedicine pursues active targeting to selectively deliver antitumor agents to cancer cells; a novel approach that promises to revolutionize therapy by dramatically increasing drug concentration in target tumor cells. In this study, we intravenously administered a liganded protein nanocarrier (T22-GFP-H6) targeting CXCR4+ lymphoma cells in mouse models to assess its selectivity as a nanocarrier by measuring its tissue biodistribution in cancer and normal cells. No previous protein-based nanocarrier has been described as specifically targeting lymphoma cells. T22-GFP-H6 achieved a highly selective tumor uptake in a CXCR4+ lymphoma subcutaneous model, as detected by fluorescent emission. We demonstrated that tumor uptake was CXCR4-dependent because pretreatment with AMD3100, a CXCR4 antagonist, significantly reduced tumor uptake. Moreover, in contrast to CXCR4+ subcutaneous models, CXCR4- tumors did not accumulate the nanocarrier. Most importantly, after intravenous injection in a disseminated model, the nanocarrier accumulated and internalized in all clinically relevant organs affected by lymphoma cells with negligible distribution to unaffected tissues. Finally, we obtained antitumor effect without toxicity in a CXCR4+ lymphoma model by administration of T22-DITOX-H6, a nanoparticle incorporating a toxin with the same structure as the nanocarrier. Hence, the use of the T22-GFP-H6 nanocarrier could be a good strategy to load and deliver drugs or toxins to treat specifically CXCR4-mediated refractory or relapsed diffuse large B-cell lymphoma without systemic toxicity.


Accuracy of phenotypic and genotypic testing for identification of Streptococcus pneumoniae and description of Streptococcus pseudopneumoniae sp. nov.
We have identified an unusual group of viridans group streptococci that resemble Streptococcus pneumoniae. DNA-DNA homology studies suggested that a subset of these isolates represent a novel species that may be included in the S. oralis-S. mitis group of viridans group streptococci. We suggest that this novel species be termed Streptococcus pseudopneumoniae. A combination of phenotypic and genetic reactions allows its identification. S. pseudopneumoniae strains do not have pneumococcal capsules, are resistant to optochin (inhibition zones, less than 14 mm) when they are incubated under an atmosphere of increased CO2 but are susceptible to optochin (inhibition zones, >14 mm) when they are incubated in ambient atmospheres, are not soluble in bile, and are positive by the GenProbe AccuProbe Pneumococcus test. The bile solubility test is more specific than the optochin test for identification of S. pneumoniae. Genetic tests for pneumolysin (ply) and manganese-dependent superoxide dismutase (sodA) and identification tests with a commercial probe, AccuProbe Pneumococcus, do not discriminate between the new species and S. pneumoniae.


Nucleotide sequence of aceK, the gene encoding isocitrate dehydrogenase kinase/phosphatase.
In Escherichia coli, the phosphorylation and dephosphorylation of isocitrate dehydrogenase (IDH) are catalyzed by a bifunctional protein kinase/phosphatase. We have determined the nucleotide sequence of aceK, the gene encoding IDH kinase/phosphatase. This gene consists of a single open reading frame of 1,734 base pairs preceded by a Shine-Dalgarno ribosome-binding site. Examination of the deduced amino acid sequence of IDH kinase/phosphatase revealed sequences which are similar to the consensus sequence for ATP-binding sites. This protein did not, however, exhibit the extensive sequence homologies which are typical of other protein kinases. Multiple copies of the REP family of repetitive extragenic elements were found within the intergenic region between aceA (encoding isocitrate lyase) and aceK. These elements have the potential for combining to form an exceptionally stable stem-loop structure (delta G = -54 kcal/mol [ca. -226 kJ/mol]) in the mRNA. This structure, which masks the ribosome-binding site and start codon for aceK, may contribute to the downshift in expression observed between aceA and aceK. Another potential stem-loop structure (delta G = -29 kcal/mol [ca. 121 kJ/mol]), unrelated to the REP sequences, was found within aceK.


The self-incompatibility (S) haplotypes of Brassica contain highly divergent and rearranged sequences of ancient origin.
In Brassica, the recognition of self-related pollen by the stigma is controlled by the highly polymorphic S locus that encodes several linked and coadapted genes and can span several hundred kilobases. We used pulsed-field gel electrophoresis to analyze the structure of different S haplotypes. We show that the S2 and S13 haplotypes of Brassica oleracea contain extensive sequence divergence and rearrangement relative to each other. In contrast, haplotypic configuration is more conserved between B. oleracea S13 and B. campestris S8, two haplotypes that have been proposed to be derived from a common ancestral haplotype based on sequence comparisons. These results support the view that extensive restructuring of the S locus preceded speciation in Brassica. This structural heteromorphism, together with haplotype-specific sequences, may suppress recombination within the S locus complex, potentially providing a mechanism for maintaining the linkage of coadapted allelic combinations of genes over time.


Immune responses in advanced colorectal cancer following repeated intradermal vaccination with the anti-CEA murine monoclonal antibody, PR1A3: results of a phase I study.
The aim was to determine the toxicity, clinical and immune responses to the murine monoclonal anti-carcinoembryonic antigen (CEA) antibody, PR1A3, in patients with advanced colorectal cancer. Fifteen patients with advanced colorectal cancer received either 0.5-, 1.0- or 5.0-mg doses of PR1A3 mixed with 10% w/v Alum adjuvant (Superfos Biosector, Denmark) intradermally at 4-week intervals for 3 months. Patient serum was assessed for anti-idiotypic (Ab2), anti-anti-idiotypic (Ab3) and human anti-mouse antibody (HAMA) reactivity. Peripheral blood mononuclear cell (PBMC) proliferation with phytohaemagglutinin (PHA), CEA and PR1A3, stimulated IL-2, IL-4 and IFN-gamma levels and PR1A3-stimulated IL-2 receptor expression during immunotherapy were determined. Comparisons were made with 16 age-matched controls without malignant disease. Hyperimmune sera from 12 of the 15 patients showed Ab2 reactivity with no detectable Ab3 responses. Strong HAMA reactivity was recorded in 7 of the 15 cases with no adverse clinical effect. Delayed-type hypersensitivity (DTH) responses developed in 12 of the 15 patients. Pre-treatment PBMC proliferation with PHA was subnormal in each patient compared with controls, becoming normal (or supranormal) in all patients during immunisation (P<0.001). PBMC proliferation with CEA and PR1A3 increased during immunotherapy (P<0.001) along with stimulated production of IL-2, IFN-gamma and IL-2 receptor expression. Progressive disease was observed in 14 of the 15 patients with minimal toxicity. PR1A3 generated limited idiotypic responses but robust DTH reactivity in most patients. In vitro PBMC proliferation with mitogens and recall antigens is greatly increased during the course of immunisation, with a shift in stimulated cytokine profile.


Management of Parthenium hysterophorus (Asteraceae) by Withania somnifera (Solanaceae).
This study was designed to evaluate the herbicidal activity of Withania somnifera (L.) Dunal against the noxious weed parthenium (Parthenium hysterophorus L.). In a laboratory bioassay, the effect of aqueous, methanol and n-hexane shoot and root extracts of 5%, 10%, 15% and 20% w/v concentrations (on a fresh weight basis) of W. somnifera were tested against the germination and seedling growth of parthenium. In general, aqueous and methanol extracts markedly suppressed the germination, root and shoot growth of parthenium. The shoot extracts were more inhibitory than the root extracts. In a foliar spray bioassay, the aqueous and methanol shoot extracts of 10% w/v (on a dry weight basis) concentration were sprayed on 1-week and 2-week-old pot-grown parthenium seedlings. Two subsequent sprays were carried out 5 and 10 days after the first spray. The aqueous and methanol extracts significantly reduced the length and biomass of parthenium shoots. In a soil amendment bioassay, the crushed shoots of W. somnifera were incorporated in the soil at 1-5% w/w. Parthenium seeds were sown one week after the residue incorporation and plants were harvested 40 days after sowing. All the soil amendment treatments significantly reduced seed germination by 43-89%. The highest dosages of 4% and 5% significantly suppressed the root and shoot biomasses of parthenium. This study concludes that foliar spray of aqueous and methanol extracts, and soil amendment with leaf residue of W. somnifera, can control the germination and growth of parthenium, one of the world's worst weeds.


Axonal TDP-43 condensates drive neuromuscular junction disruption through inhibition of local synthesis of nuclear encoded mitochondrial proteins.
Mislocalization of the predominantly nuclear RNA/DNA binding protein, TDP-43, occurs in motor neurons of ~95% of amyotrophic lateral sclerosis (ALS) patients, but the contribution of axonal TDP-43 to this neurodegenerative disease is unclear. Here, we show TDP-43 accumulation in intra-muscular nerves from ALS patients and in axons of human iPSC-derived motor neurons of ALS patient, as well as in motor neurons and neuromuscular junctions (NMJs) of a TDP-43 mislocalization mouse model. In axons, TDP-43 is hyper-phosphorylated and promotes G3BP1-positive ribonucleoprotein (RNP) condensate assembly, consequently inhibiting local protein synthesis in distal axons and NMJs. Specifically, the axonal and synaptic levels of nuclear-encoded mitochondrial proteins are reduced. Clearance of axonal TDP-43 or dissociation of G3BP1 condensates restored local translation and resolved TDP-43-derived toxicity in both axons and NMJs. These findings support an axonal gain of function of TDP-43 in ALS, which can be targeted for therapeutic development.


Power of averaging: Noise reduction by ensemble coding of multiple faces.
Face perception is an important ability in social interaction. Holistic processing of a face enables us to understand facial attributes easily despite similarity of faces. Ensemble coding of multiple faces enables us to judge the characteristics of a crowd easily despite the limited capacity of the visual system. The current study investigated whether holistic processing of a face is different from ensemble coding of multiple faces and, if so, how. We tested how face inversion influenced both holistic processing and ensemble coding. Participants were asked to discriminate emotion intensities of either a single face or a crowd of faces (angry or happy face[s]). More important, face(s) or facial features were sometimes presented upside-down. We found that a face inversion effect was larger in discriminating individual emotion than in discriminating emotion in a crowd. As the number of inverted components increased, both discrimination abilities were more disrupted. The degree of disruption varied depending on different emotions and features in holistic processing, but it did not vary in ensemble coding. The inversion effect was weaker in ensemble coding than in holistic processing because noise produced by inversion might have cancelled out in averaging. Different inversion effects depending on different emotions and facial features were observed only in holistic processing because uneven contributions of facial features to individual emotion processing might have reduced the effectiveness of noise cancellation. Together, these results suggest that ensemble coding provides a powerful mechanism of noise cancellation involved with individual representation. (PsycINFO Database Record (c) 2020 APA, all rights reserved).


Predictors and Prognostic Implications of Incident Heart Failure in Patients With Prevalent Atrial Fibrillation.
The purpose of this study was to determine the significant clinical predictors of incident heart failure (HF) and its prognostic effect on long-term outcomes among community-based patients with established atrial fibrillation (AF). AF is associated with an increased risk of HF. However, in this population, little focus is placed on risk stratification for and the prevention of HF. Patients with AF but without HF at baseline enrolled in the ORBIT-AF (Outcomes Registry for Informed Treatment of Atrial Fibrillation) registry were included. Separate multivariable-adjusted Cox frailty regression models were used to identify significant predictors of HF incidence and determine the associated risk of adverse clinical events. The study included 6,545 participants with AF from 173 participating sites. Incident HF developed in 236 participants (3.6%) over the 2-year follow-up period; ejection fraction was preserved (>40%) in 64%, reduced (≤40%) in 13.5%, and missing in 22.5%. In multivariable analysis, traditional HF risk factors (age, coronary artery disease, renal dysfunction, and valvular disease), presence of permanent AF (hazard ratio [HR]: 1.60 [95% confidence interval (CI): 1.18 to 2.16]; reference group: paroxysmal AF), and elevated baseline heart rate (HR: 1.07 [95% CI: 1.02 to 1.13] per 5 beats/min higher heart rate) were independently associated with incident HF risk. Incident HF among patients with AF was independently associated with higher risk of mortality, all-cause hospitalization, and bleeding events. Incident HF among patients with AF is common, is more likely to be HF with preserved ejection fraction, and is associated with poor long-term outcomes. Traditional HF risk factors, AF type, and baseline heart rate are independent clinical predictors of incident HF.


A cross-cultural study of reactivation of posttraumatic stress disorder symptoms: American and Cambodian psychophysiological response to viewing traumatic video scenes.
A physiological hyperarousal state, which can be reactivated by traumatic stimuli, occurs frequently in patients with posttraumatic stress disorder (PTSD). The goals of this study were to determine whether physiological hyperarousal measured by increased heart rate is a specific response to reminders of a patient's own traumatic events or a more generalized hyperarousal state. Five brief videotape scenes of traumatic events (hurricane, auto accident, Cambodian refugee camp, domestic violence, and Vietnam War) were shown to two patient groups with PTSD (Vietnam veterans and Cambodian refugees) and three control groups (Vietnam veterans, Cambodian refugees, and nonpatient Americans). Observations of subjects' behavior, subjective ratings of distress, and heart rate change were recorded and evaluated. The results indicated that Cambodians with PTSD had the most reactions as measured by behavior and heart rate changes. These tended to occur during all scenes, not just the specific Cambodian scene, indicating a general nonspecific arousal. The Vietnam veterans had the fewest changes implying an inhibition of response. The control groups were intermediate in physiological response. The response in PTSD patients to reactivation scenes is complex and probably relates to type and degree of trauma, as well as to culture.


In situ responses of the sponge microbiome to ocean acidification.
Climate change is causing rapid changes in reef structure, biodiversity, and function, though most sponges are predicted to tolerate conditions projected for 2100. Sponges maintain intimate relationships with microbial symbionts, with previous studies suggesting that microbial flexibility may be pivotal to success under ocean acidification (OA). We performed a reciprocal transplantation of the coral reef sponges Coelocarteria singaporensis and Stylissa cf. flabelliformis between a control reef site and an adjacent CO2 vent site in Papua New Guinea to explore how the sponge microbiome responds to OA. Microbial communities of C. singaporensis, which differed initially between sites, did not shift towards characteristic control or vent microbiomes, even though relative abundances of Chloroflexi and Cyanobacteria increased and that of Thaumarchaeota decreased 7 months after transplantation to the control site. Microbial communities of S. cf. flabelliformis, which were initially stable between sites, did not respond specifically to transplantation but collectively exhibited a significant change over time, with a relative increase in Thaumarchaeota and decrease in Proteobacteria in all treatment groups. The lack of a community shift upon transplantation to the vent site suggests that microbial flexibility, at least in the adult life-history stage, does not necessarily underpin host survival under OA .


Changes in intrinsic subtype of breast cancer during tumor progression in the same patient.
Hormone receptor (HR), human epidermal growth factor receptor 2 (HER2) and Ki67 are important prognostic factors and key variables in classification of the intrinsic subtype, which is essential for choice of adjuvant therapy in breast cancer management. There has been earlier reports that instability of hormonal and HER2 status during progression of tumor. However, breast cancer treatment guidelines recently recommended using the intrinsic subtype that is determined by four immunohistochemical (IHC) assays, estrogen receptor (ER), progesterone receptor (PR), HER2 and Ki67. The purpose of study was to investigate whether the intrinsic subtype changes during the tumor progression from ductal carcinoma in situ (DCIS) to lymph node metastasis. The study included 90 patients with breast cancer in Korea University Guro Hospital, between 1992 and 2008. All individuals had DCIS, invasive carcinoma and lymph node metastasis lesion. IHC staining for ER, PR, HER2 and Ki67 as well as SISH assay for HER2 gene amplification was done with following standard method. Overall 25% of breast cancer changed their intrinsic phenotype during progression. Study demonstrated that a subset of breast cancers can change their intrinsic subtype during cancer progression. These changes have an impact on patient prognosis and management, because each breast cancer subtype has their own differently optimized treatment options according to St. Gallen and NCCN guideline.


Effects of the multiple-piece maxillary osteotomy on the periodontium.
Clinical indications exist for both the surgically assisted rapid maxillary expansion (SARME) and the multiple-piece maxillary osteotomy (MPMO). Recent trends, however, imply that the SARME combined with a subsequent 1-piece osteotomy can supplant the use of the MPMO. Those favoring the SARME frequently site morbidities associated with the MPMO. Major reported complications include loss of dentoalveolar segments, teeth, and oronasal or oroantral communication. Relapse, tooth devitalization, and damage to the periodontium, including bone loss and soft tissue alteration, comprise the minor morbidities. If these can be avoided or minimized, then the use of the MPMO for its inherent advantages over the SARME in certain clinical situations may be indicated The purpose of our study was to critically evaluate the periodontium following the use of the MPMO to ascertain if minor morbidities are inherent to the procedure, and to quantify them. Records of 24 MPMO patients were reviewed, ranging from 3 to 24 months after surgery. A specific surgical technique was utilized for all patients, including bone grafting. The vertical segmental osteotomy sites varied and were recorded for comparison. Periodontal probing depths at the segmental osteotomy sites were compared with the adjacent interproximal spaces of each patient. Independent dental examiners were used to review photographs and periapical radiographs to compare the papillae and alveolar bone height, respectively, at the osteotomy site versus the neighboring interproximal areas. A paired t test was used to compare probing depth measurements at the vertical osteotomy site and neighboring interproximal sites. The mean difference between these two sites was 0.01 mm with a standard deviation of 0.25 mm. This was not statistically significant. Statistical analyses were also performed to compare these probing depth differences at varying sites in the maxilla, and to compare probing depth differences to gender, total number of osteotomies performed on each patient, estimated blood loss, and length of procedure. These results were not statistically significant. Independent examiners found no difference in gingival architecture or alveolar bone levels when comparing vertical osteotomy sites to neighboring interproximal sites. This study showed that damage to the periodontium at vertical osteotomy sites was minimal, and not a reason to avoid use of the multiple-piece maxillary osteotomy.


Attentional and perceptual asymmetries in an immersive decision-making task.
Pseudoneglect represents the tendency in healthy people to show a slight bias in favour of stimuli appearing in the left visual field. Some studies have shown that this leftward bias can be annulled or reserved towards a rightward bisection bias when lateral attentional biases are assessed in far space. Using an immersive simulated, ecologically valid football task, we investigated whether possible attentional and perceptual asymmetries affect sport-specific decision making. Twenty-seven sport athletes were required to judge different game situations, which involved both perceptual and attentional skills to perceive player configurations in the visual periphery. We did not find any performance differences in accuracy rate between the left and right visual field side for stimuli presented close to the screen centre in an object-detection (perception-based) and feature-recognition (attention-based) task. This result is in line with previous findings showing an absence of a left- or rightward bisection bias in far space. However, accuracy was higher for stimuli being presented at visual angles wide away from the screen centre at the left side compared to the right side of visual field. This finding cannot be explained by literature focusing on pseudoneglect in far space, but rather by previous findings on landmark judgments often showing left bias both in near and in far space. Overall, the current findings provide new perspectives on attentional and perceptual asymmetries in real-world scenarios, and different interpretations of results are discussed.


Small and dim target detection via lateral inhibition filtering and Artificial Bee colony based selective visual attention.
This paper proposed a novel bionic selective visual attention mechanism to quickly select regions that contain salient objects to reduce calculations. Firstly, lateral inhibition filtering, inspired by the limulus' ommateum, is applied to filter low-frequency noises. After the filtering operation, we use Artificial Bee Colony (ABC) algorithm based selective visual attention mechanism to obtain the interested object to carry through the following recognition operation. In order to eliminate the camera motion influence, this paper adopted ABC algorithm, a new optimization method inspired by swarm intelligence, to calculate the motion salience map to integrate with conventional visual attention. To prove the feasibility and effectiveness of our method, several experiments were conducted. First the filtering results of lateral inhibition filter were shown to illustrate its noise reducing effect, then we applied the ABC algorithm to obtain the motion features of the image sequence. The ABC algorithm is proved to be more robust and effective through the comparison between ABC algorithm and popular Particle Swarm Optimization (PSO) algorithm. Except for the above results, we also compared the classic visual attention mechanism and our ABC algorithm based visual attention mechanism, and the experimental results of which further verified the effectiveness of our method.


Effects of lead on cholinergic SN56 neuroblastoma cells.
Age-dependent accumulation of lead in brain has been implicated in the pathomechanisms of Alzheimer's disease. The aim of this work was to investigate whether cholinotoxic effects of lead may result from alterations in acetyl-CoA metabolism. One day exposure of differentiated SN56 cholinergic neuroblastoma cells to 0.5 micromol/L lead or 0.01 mmol/L amyloid-beta1-42, increased fraction of nonviable cells to about 2%. Suppression of choline acetyltransferase activity occurred only in the presence of fresh amyloid-beta1-42, whereas lead was ineffective. All agents in combination caused suppression of acetyl-CoA in cytoplasm and mitochondria down to 19% and 34% of controls, respectively. Inverse correlation was observed between whole cell acetyl-CoA level and fraction of nonviable cells at different combinations of lead and other neurotoxic compounds. It indicates that lead had no primary suppressive effect on cholinergic phenotype but, at least in part, exerted cytotoxic influence on cholinergic neurons through the decrease of their acetyl-CoA.


Use of a handheld terahertz pulsed imaging device to differentiate benign and malignant breast tissue.
Since nearly 20% of breast-conserving surgeries (BCS) require re-operation, there is a clear need for developing new techniques to more accurately assess tumor resection margins intraoperatively. This study evaluates the diagnostic accuracy of a handheld terahertz pulsed imaging (TPI) system to discriminate benign from malignant breast tissue ex vivo. Forty six freshly excised breast cancer samples were scanned with a TPI handheld probe system, and histology was obtained for comparison. The image pixels on TPI were classified using (1) parameters in combination with support vector machine (SVM) and (2) Gaussian wavelet deconvolution in combination with Bayesian classification. The results were an accuracy, sensitivity, specificity of 75%, 86%, 66% for method 1, and 69%, 87%, 54% for method 2 respectively. This demonstrates the probe can discriminate invasive breast cancer from benign breast tissue with an encouraging degree of accuracy, warranting further study.


Testing a Self-Affirmation Intervention for Improving the Psychosocial Health of Black and White Medical Students in the US.
Self-affirmation interventions have been shown to mitigate the negative psychological effects of stereotype threat on Black students in secondary and undergraduate education. However, there is currently limited research testing whether Black students in medical schools may also experience the negative influences of stereotype threat. Until now, it has been unclear whether Black (vs. White) students experience a lower sense of belonging in medical school and whether they can benefit from self-affirmation interventions during medical training. With a longitudinal field experiment, we tested (a) whether Black (vs. White) medical students in the US experience decrements in psychological well-being (i.e., fatigue, depression, anxiety), sense of belonging, perceived residency competitiveness, and residency goal stability; and (b) the extent to which a self-affirmation intervention would ameliorate any observed disparities in these outcomes for Black students. With a sample of 234 Black and 182 White medical students across 50 schools in the US, we found that Black students tended to report more fatigue and less belonging than White students; however, the self-affirmation intervention did not significantly influence students' fatigue, depression, anxiety, or belonging. Unexpectedly, Black students in the self-affirmation (vs. control) condition reported lower perceived competitiveness for residency. White students' perceived competitiveness for residency was unaffected by the intervention. Exploratory analyses revealed that Black (vs. White) students were less likely to indicate stable residency goals over time, which may be an indication of threat; however, this racial gap was eliminated with the intervention. We discuss the plausible reasons for these findings and provide recommendations for future work in this area.


Thoracoabdominal Asynchrony Is Not Associated with Oxyhemoglobin Saturation in Recovering Premature Infants.
Recovering premature infants are at risk for hypoxemia and lack of synchrony between their rib cage and abdomen due to airflow obstruction and poor respiratory compliance. Thoracoabdominal asynchrony (TAA) is a useful marker of resistive and elastic lung properties. Whether TAA predicts oxygenation is unknown. We investigated oxyhemoglobin saturation (SpO2%) and TAA (phase angle, φ) in preterm infants with/without high-humidity nasal cannula (HHNC). A cross-sectional observational study was conducted in 92 infants at 32 weeks' postmenstrual age. We measured SpO2% with pulse oximetry and TAA with φ via respiratory inductance plethysmography in infants (mean gestational age: 26.4 + 1.3 weeks) who required room air (n = 18) or HHNC with/without supplemental oxygen (1-5 liters per minute, FiO2 0.21-0.33, n = 74). We calculated median SpO2% from 24.7 + 10.0 min of quiet sleep and median φ from up to 60 breaths. Infants breathing room air alone had marked TAA (φ = 83.6 + 32.9°, range: 10.9-148.5) as did those receiving varying degrees of ventilatory and oxygen support via HHNC (range of group means, φ = 47.0-90.0°). Infants breathing room air had statically greater median SpO2% than those receiving support (96.3 + 0.6% vs. 91.3 + 0.6%; ANOVA p = 0.001). SpO2% was not associated with TAA in either group (r2 = 0.09). Recovering premature infants exhibited TAA regardless of need for ventilatory support and supplemental oxygen. TAA was not associated with SpO2% in either group. Maintenance of SpO2% does not require correction of TAA.


Symptom cluster, healthcare use and mortality in patients with severe chronic obstructive pulmonary disease.
To examine how subgroups of patients with chronic obstructive pulmonary disease, identified by ratings of symptoms (dyspnoea, anxiety, depression and fatigue), affect healthcare use and mortality. People with chronic obstructive pulmonary disease often experience multiple symptoms. The importance of multiple symptoms and symptom clusters has received increased attention. However, little is known about symptom clusters and their effect on healthcare use and mortality in this population. Descriptive cross-sectional study. This secondary data analysis used data from the National Emphysema Treatment Trial. Participants (n = 597) had severe chronic obstructive pulmonary disease. Descriptive and inferential statistics were used to analyse the data that were drawn from structured interviews, questionnaires and clinical measures. Three subgroup clusters emerged based on four symptom ratings. Mean age, proportion with higher education, proportion using oxygen, disease severity, exercise capacity and quality of life differed significantly between subgroups. Participants with high levels of symptoms used healthcare services more and were more likely to have died at the five-year follow-up than those with low levels of symptoms. Symptom cluster subgroups had more significant relationship with mortality than single symptoms. Patients with high levels of symptoms require greater clinical attention. Understanding subgroups of patients, based on symptom ratings and their adverse effect on outcomes, may enable healthcare providers to assess multiple symptoms and identify subgroups of patients at risk of increased healthcare use and mortality. Targeting modifiable symptoms within the cluster may be more beneficial than focusing on a single symptom for certain health-related outcome.


Endogenous opioids and baroreflex control in humans.
We describe two studies designed to elucidate the role of endogenous opioids in blood pressure control in humans. In the first study, nine normal subjects received infusions of DAMME (a metenkephalin analog), naloxone, or saline, and blood pressure, heart rate, and plasma norepinephrine concentration were determined supine and following 5 minutes of 70 degrees head-up tilt at intervals for 6 hours. Blood pressure following tilt was significantly decreased by DAMME but not influenced by naloxone, the effect being most marked at 3 hours (placebo = 110 +/- 6/78 +/- 7 mm Hg; naloxone = 106 +/- 10/79 +/- 5 mm Hg; DAMME = 96 +/- 16/67 +/- 8 mm Hg (p less than 0.01). However, heart rate and plasma norepinephrine did not rise in response to this hypotension. Heart rates at 3 hours were: placebo = 87 +/- 16 bpm; naloxone = 88 +/- 19 bpm; DAMME = 89 +/- 23 bpm. Plasma norepinephrine levels (nmol/liter) at 3 hours were: placebo = 6.0 +/- 2.2; naloxone = 5.8 +/- 1.9; DAMME = 6.0 +/- 1.9. In the second study, seven normal subjects had blood pressure reduced by incremental infusions of sodium nitroprusside, and the effects of placebo, naloxone, and DAMME on the slope of the heart period/blood pressure relationship investigated. Naloxone significantly increased the slope by 90% and DAMME significantly reduced the slope by 30%. It is concluded that endogenous opioids modulate the baroreflex control of blood pressure in normal humans.


Relocation of remote dwellers living with hemodialysis: a time trade-off survey.
There has been little research exploring the experience of dialysis therapy for people living in remote communities. Remote residence location has previously been associated with excess mortality in hemodialysis (HD) patients, suggesting that relocation to a referral center might improve outcomes. It is unknown whether patients view this approach as acceptable. We studied 121 remote-dwelling chronic HD patients using the time trade-off method applied to hypothetical scenarios. Participants indicated that they would trade a median of 6 years of life in their current location (including current social supports) (95% CI 2.25-7) for 10 years of life in a referral center without any of their existing social supports (meaning they would be willing to forgo 4 years of life to remain in their current residence location). When current social supports were assumed to continue in both locations, people were only willing to forego a median of 2 years of life (95% CI 1-4) to remain in their current location. Older participants were much less willing to accept relocation than younger participants; the median time trade-off associated with relocation and without social supports was 2 years for participants aged <50 years, 3 years for those aged 50-69.9 years and 9 years for those aged ≥70 years. Hemodialysis patients currently living remotely were willing to forgo much of their remaining life expectancy rather than relocate-especially among older participants. These findings suggest that decisions about relocation should be accompanied by discussion of anticipated changes in quality of life and life expectancy.


Localization of estrogen receptors in interstitial cells of hamster kidney and in estradiol-induced renal tumors as evidence of the mesenchymal origin of this neoplasm.
The mechanism of estrogen-induced and -dependent kidney carcinogenesis in Syrian hamsters and the cell of origin of the tumor are not well understood; they have been investigated in this study by mapping the cellular locations of estrogen receptor (ER) in estrogen-dependent tumors, in kidney tissue of hamsters treated with estradiol for 0.5 and 5.5 months, and in kidneys of age-matched controls. To validate the methods used, receptors have also been localized in uteri of hamsters and rats and in female hamster kidneys. ERs have been identified in cryostat sections by immunocytochemical techniques using an affinity-purified ER antibody, ER-715. Nuclei of tumors were intensely stained for ERs. In estrogen-treated kidneys and in controls, ER protein was identified in interstitial cells and capillaries, in arteries, and in renal corpuscles, particularly in podocytes and in the parietal layers surrounding the renal corpuscles. There was no ER protein in tubular epithelia even when tubuli were surrounded by tumor cells. The ER distribution in female hamster kidneys closely matched that in male kidneys. However, the staining intensity was stronger in female than in male kidneys. In hamster uteri, there was an intense ER-positive reaction in the nuclei of stroma, in stromal vessels, and in the luminal epithelia as demonstrated previously by others in rat uteri. ER mRNA has also been demonstrated by Northern blot analysis in estrogen-treated kidneys which contained tumors but was undetectable in untreated kidneys. The localization of ERs in estrogen-dependent tumors and in interstitial cell types but not in tubular epithelia supports previous conclusions of an interstitial origin of estrogen-induced hamster kidney tumors.


New insights into human papillomavirus-associated head and neck squamous cell carcinoma.
Human papillomavirus (HPV)-associated head and neck squamous cell carcinoma (HNSCC) is an entity with peculiar clinical and molecular characteristics, which mainly arises from the reticulated epithelium lining the crypts of the palatine tonsils and the base of the tongue. The only head and neck site with a definite etiological association between persistent high-risk (HR) HPV infection and development of SCC is the oropharynx. HPV-positive malignancies represent 5-20% of all HNSCCs and 40-90% of those arising from the oropharynx, with widely variable rates depending on the geographic area, population, relative prevalence of environment-related SCC and detection assay. HPV-16 is by far the most common HR HPV genotype detected in oropharyngeal SCC (OPSCC), and the only definitely carcinogenic genotype for the head and neck region. Patients with HPV-induced OPSCC are more likely to be middle-aged white men, non-smokers, non-drinkers or mild to moderate drinkers, with higher socioeconomic status and better performance status than subjects with HPV-unrelated SCC. HPV-induced HNSCCs are often described as non-keratinizing, poorly differentiated or basaloid carcinomas, and are diagnosed in earlier T-category with a trend for a more advanced N-category, with cystic degeneration, than the HPV-unrelated carcinomas. HPV positivity is associated with better response to treatment and modality-independent survival benefit. Treatment selection in HPV-related oropharyngeal carcinoma is becoming a critical issue, and although there is no evidence from randomized, controlled trials to support a treatment de-escalation in HPV-positive SCC, some investigators argue that intensive combined modality strategies may represent an overtreatment.


International distance education and the transition from ISDN to high-bandwidth Internet connectivity.
The Thailand-Hawaii Assessment of Interactive Healthcare Initiative (THAI-HI) is an international distance-education project between two teaching hospitals in Honolulu and Bangkok that uses videoconferencing over three ISDN lines. A 'morning report' format is used to discuss clinical cases primarily covering infectious disease and critical-care topics. An audience response system is used at both sites to add interactivity. From July 2001 to May 2002, 816 health-care providers attended 20 clinical conferences. Audiences rated the conferences as highly relevant and as having high training value. Since the ISDN connection is expensive, we plan to convert the telecommunications to a high-bandwidth Internet connection. The Honolulu site will use a 45 Mbit/s commercial connection to the Hawaii Intranetwork Consortium, which links to the Abilene Network on the US mainland. The Bangkok hospital will use a 155 Mbit/s wireless optical connection to UNINET Thailand, which has a 45 Mbit/s circuit to Abilene.


The mitochondrial contact site complex, a determinant of mitochondrial architecture.
Mitochondria are organelles with a complex architecture. They are bounded by an envelope consisting of the outer membrane and the inner boundary membrane (IBM). Narrow crista junctions (CJs) link the IBM to the cristae. OMs and IBMs are firmly connected by contact sites (CS). The molecular nature of the CS remained unknown. Using quantitative high-resolution mass spectrometry we identified a novel complex, the mitochondrial contact site (MICOS) complex, formed by a set of mitochondrial membrane proteins that is essential for the formation of CS. MICOS is preferentially located at the CJs. Upon loss of one of the MICOS subunits, CJs disappear completely or are impaired, showing that CJs require the presence of CS to form a superstructure that links the IBM to the cristae. Loss of MICOS subunits results in loss of respiratory competence and altered inheritance of mitochondrial DNA.


Topology of synaptic connectivity constrains neuronal stimulus representation, predicting two complementary coding strategies.
In motor-related brain regions, movement intention has been successfully decoded from in-vivo spike train by isolating a lower-dimension manifold that the high-dimensional spiking activity is constrained to. The mechanism enforcing this constraint remains unclear, although it has been hypothesized to be implemented by the connectivity of the sampled neurons. We test this idea and explore the interactions between local synaptic connectivity and its ability to encode information in a lower dimensional manifold through simulations of a detailed microcircuit model with realistic sources of noise. We confirm that even in isolation such a model can encode the identity of different stimuli in a lower-dimensional space. We then demonstrate that the reliability of the encoding depends on the connectivity between the sampled neurons by specifically sampling populations whose connectivity maximizes certain topological metrics. Finally, we developed an alternative method for determining stimulus identity from the activity of neurons by combining their spike trains with their recurrent connectivity. We found that this method performs better for sampled groups of neurons that perform worse under the classical approach, predicting the possibility of two separate encoding strategies in a single microcircuit.


Life cycle assessment study on polishing units for use of treated wastewater in agricultural reuse.
A life cycle assessment (LCA) approach was used in the assessment of environmental impacts of some polishing units for reuse of wastewater treatment plant effluents in agricultural irrigation. These alternative polishing units were assessed: (1) microfiltration and ultraviolet (UV) disinfection, (2) cartridge filter and ultrafiltration (UF), and (3) just UV disinfection. Two different energy sources, electric grid mix and natural gas, were considered to assess the environmental impacts of them. Afterwards, the effluent of each case was evaluated against the criteria required for irrigation of sensitive crops corresponding to Turkey regulations. Evaluation of environmental impacts was carried out with GaBi 6.1 LCA software. The overall conclusion of this study is that higher electricity consumption causes higher environmental effects. The results of the study revealed that cartridge filter and UF in combination with electric grid mix has the largest impact on the environment for almost all impact categories. In general, the most environmentally friendly solution is UV disinfection. The study revealed environmental impacts for three alternatives drawing attention to the importance of the choice of the most appropriate polishing processes and energy sources for reuse applications.


Impact of baseline C-reactive protein levels on the response to secukinumab in ankylosing spondylitis: 3-year pooled data from two phase III studies.
To evaluate the magnitude of response to secukinumab treatment over 3 years in patients with ankylosing spondylitis (AS) grouped by baseline C-reactive protein (CRP) levels in a pooled study of two pivotal phase III studies: MEASURE 1 (NCT01358175) and MEASURE 2 (NCT01649375). This post hoc analysis pooled data from all patients with available baseline CRP in the two studies who received subcutaneous secukinumab 150 mg (approved dose; N=197) or placebo (N=195). Assessed efficacy endpoints included Assessments of SpondyloArthritis international Society (ASAS)20/40, Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), BASDAI50, AS Disease Activity Score inactive disease and ASAS partial remission among patients grouped by baseline CRP based on central laboratory cut-off <5 mg/L (normal) or ≥5 mg/L (elevated) and a cut-off <10 mg/L or ≥10 mg/L. At baseline, 36.5% (143/392) patients had normal and 63.5% (249/392) had elevated CRP. At week 16, ASAS20/40 response rates were higher for secukinumab versus placebo in normal (56.9%/34.7% vs 28.2%/7.0%; p<0.01/p<0.001) and in elevated (63.2%/42.4% vs 29.0%/15.3%; both p<0.0001) CRP groups. Improvement was reported for all outcomes (p<0.05) in both groups, except for ASAS partial remission in the normal CRP group, where a numerical difference 12.5% vs 2.8%, p=0.07) was observed. Similar trends of improvement were observed in the <10 and ≥10 mg/L groups across all efficacy outcomes at week 16. Treatment responses to secukinumab in all CRP groups further improved over 156 weeks. Secukinumab 150 mg demonstrated rapid and sustained efficacy in patients with AS irrespective of baseline CRP, with greater magnitude of response in patients with more elevated CRP.


UV-B damages eyes of barnacle larvae and impairs their photoresponses and settlement success.
The impact of enhanced UV-B radiation on marine ecosystems due to ozone depletion has caused growing global concern. Barnacle larvae have evolved complex photoreceptors and elaborate phototactic behaviors, which enable them to identify suitable habitats for feeding and settlement. For the first time, we demonstrate that environmentally realistic levels of UV-B radiation can induce ocular damage in barnacle larvae, thereby impairing the phototactic behavior of naupliar larvae and reducing settlement success of cypris larvae. Significant disruptions of rhabdomeres (the photosensitive structures in which phototransduction takes place) occurred in the retinular cells of naupliar eyes when naupliar larvae were exposed to a UV-B dose of 7.2 kJ m(-2), and impairment was dependent upon dose rather than irradiance. Our experimental data also showed that phototaxis of nauplii was ca. 4 times more sensitive to UV-B than settlement of cyprids. Since barnacles play an important role in the function and structure of coastal systems worldwide, any impairment of phototactic and settlement behavior of the larvae would pose a significant threat to the sustainability of this ecologically important species. The fact that enhanced UV-B radiation can induce ocular damage and subsequent phototactic impairment in barnacle larvae suggests that UV-B may also cause similar damage to other zooplankton species.


Spatial approximation between the amino terminus of a peptide agonist and the top of the sixth transmembrane segment of the secretin receptor.
Distinct spatial approximations between residues within the secretin pharmacophore and its receptor can provide important constraints for modeling this agonist-receptor complex. We previously used a series of probes incorporating photolabile residues into positions 6, 12, 13, 14, 18, 22, and 26 of the 27-residue peptide and demonstrated that each covalently labeled a site within the receptor amino terminus. Although supporting a critical role of this domain for ligand binding, it does not explain the molecular mechanism of receptor activation. Here, we developed probes having photolabile residues at the amino terminus of secretin to explore possible approximations with a different receptor domain. The first probe incorporated a photolabile p-benzoyl-l-phenylalanine into the position of His(1) of rat secretin ([Bpa(1),Tyr(10)]secretin-27). Because His(1) is critical for function, we also positioned a photolabile Bpa as an amino-terminal extension, in positions -1 (rat [Bpa(-1),Tyr(10)]secretin-27) and -2 (rat [Bpa(-2),Gly(-1),Tyr(10)]secretin-27). Each analog was shown to be a full agonist, stimulating cAMP accumulation in receptor-bearing Chinese hamster ovary-SecR cells in a concentration-dependent manner, with the position -2 probe being most potent. They bound specifically and saturably, although the position 1 analog had lowest affinity, and all were able to label the receptor efficiently. Sequential specific cleavage, purification, and sequencing demonstrated that the sites of covalent attachment for each probe were high within the sixth transmembrane segment. This suggests that secretin binding may exert tension between the receptor amino terminus and the transmembrane domain to elicit a conformational change effecting receptor activation.


Heterogeneity in disease severity in a family with a novel G68V GCK activating mutation causing persistent hyperinsulinaemic hypoglycaemia of infancy.
Glucokinase (GCK)-activating mutations cause persistent hyperinsulinaemic hypoglycaemia of infancy (PHHI). GCK-PHHI patients have regulated insulin secretion and can usually be treated with diazoxide. The six reported cases suggest that the severity of the mutation predicts the clinical phenotype. The aim of this study was to relate genotype to phenotype [clinical phenotype, glucose-stimulated insulin release (GSIR) and GCK functional analysis] in a large pedigree with eight affected individuals. The genes encoding B-cell GCK and the K(ATP) channel subunits (ABCC8 and KCNJ11) were sequenced to identify mutations for functional analysis. Genetic variants influencing B-cell function were genotyped in affected individuals. Islet secretory capacity was determined by oral glucose tolerance test A novel GCK mutation (G68V) co-segregating with hypoglycaemia was identified in eight family members. Kinetic analysis revealed that G68V-GCK activity is ~16 times more than wild-type-GCK with an increased affinity for glucose [concentration at half maximal activation (S(0.5)) 1.94 +/- 0.16 vs. 7.43 +/- 0.12, mutant vs. wild type, mean +/- sem]. Mathematical modelling predicted a threshold for GSIR of 1.9 mmol/l in the mutant. Oral glucose tolerance tests showed regulated insulin secretion. The severity of hypoglycaemia and related symptoms in affected subjects were heterogeneous. Clinical presentations were asymptomatic (n = 1), extreme hunger (n = 3), seizures (n = 2) and loss of consciousness (n = 2); 7/8 were managed with diet but the proband was treated with diazoxide and octreotide. Phenotypic modification by a second mutation in the K(ATP) channel genes (ABCC8, KCNJ11) or by common genetic variants in KCNJ11, GCK and TCF7L2 was excluded. The novel activating GCK mutation G68V is associated with variable phenotypic severity, supporting modification of GSIR by genetic and/or environmental factors.


A magnesium-dependent mreB null mutant: implications for the role of mreB in Bacillus subtilis.
MreB shares a common prokaryotic ancestor with actin and is present in almost all rod-shaped bacteria. MreB proteins have been implicated in a range of important cell processes, including cell morphogenesis, chromosome segregation and cell polarity. The mreB gene frequently lies at the beginning of a cluster of genes, immediately upstream of the conserved mreC and mreD genes. RNA analysis showed that in Bacillus subtilis mreB is co-transcribed with mreC and that these genes form part of an operon under the control of a promoter(s) upstream of mreB. Construction of an in-frame deletion of mreB and its complementation by mreB(+) only, in trans, established that the gene is important for maintenance of cell width and cell viability under normal growth conditions, independent of polar effects on downstream genes. Remarkably, virtually normal growth was restored to the mreB null mutant in the presence of high concentrations of magnesium, especially when high concentrations of the osmoprotectant, sucrose were also present. Under these conditions, cells could be maintained in the complete absence of an mreB gene, with almost normal morphology. No detectable effect on chromosome segregation was evident in the mutant, nor was there an effect on the topology of nascent peptidoglycan insertion. A GFP-MreB fusion was used to look at the localization of MreB in live cells. The pattern of localization was similar to that previously described, but no tight linkage to nucleoid positioning was evident. Propagation of the mreB null mutant in the absence of magnesium and sucrose led to a progressive increase in cell width, culminating in cell lysis. Cell division was also perturbed but this effect may be secondary to the disturbance in cell width. These results suggest that the major role of MreB in B. subtilis lies in the control of cell diameter.


Aging among elite distance runners: a 22-yr longitudinal study.
The purpose of this study was to assess the physiological responses of former elite distance runners during submaximal and maximal exercise after a mean period of 22 yr. Fifty-three men were initially tested (T1) in the late 1960s and early 1970s when they were all highly trained and competitive. For the current evaluation (T2), these men were classified as highly trained (HT; n = 10), fitness trained (FT; n = 18), untrained (UT; n = 15), and fit older (FO; n = 10), depending on their continued level of training and age. The mean (+/- SE) age for the HT, FT, and UT men during T2 was similar (46.5 +/- 1.6 yr), whereas the FO men were significantly (P < 0.05) older (68.4 +/- 2.7 yr). All groups experienced a significant decrease (P < 0.05) in maximal O2 uptake (VO2 max) from T1 to T2. However, this decrease was related to the amount of training between evaluations. The HT men had the smallest reduction (6% per decade) in VO2 max (from 68.8 to 59.2 ml.g-1.min-1). The FT men's VO2 max was approximately 10% lower per decade (from 64.1 to 48.9 ml.kg-1.min-1), whereas an approximately 15% decrease per decade was observed for the UT (from 70.7 to 46.7 ml.kg-1.min-1) and FO (from 60.3 to 40.7 ml.kg-1.min-1) men, despite the continued training of the FO men. Energy requirements for a standardized run at 12 km/run were similar from T1 to T2 for the HT and FT men, whereas the UT men required an increased (P < 0.05) O2 uptake (40.3-41.8 l/min), ventilation (53.7-72.7 l/min), and heart rate (127-142 beats/min). The perceived effort and %VO2 max for this submaximal run were greater during T2 for all groups, which was related to the decline in VO2 max. These longitudinal data indicate that after more than two decades the physiological capacities of these aging runners are compromised, regardless of training. These data also confirm previous cross-sectional findings that aerobic capacity of highly trained middle-aged men declines approximately 5-7% per decade.


A Framework for Treatment Decision Making at Prostate Cancer Recurrence.
Of the 50,000 men in the US who elect for radical prostatectomy for prostate cancer, 24% to 40% will have a prostate-specific antigen (PSA) recurrence (PSA-R) within 10 years. Deciding whether to administer salvage therapy (ST) at PSA-R presents challenges, as treatment reduces the risk of progression to clinical metastasis but incurs unnecessary side effects should the man die before metastasis. We have developed a new harm-benefit framework using a clinical cohort to inform shared decision making between patients and physicians at PSA-R. Records of 1,045 Johns Hopkins University Hospital patients diagnosed between 1984 and 2013 who had PSA-R following radical prostatectomy were analyzed using marginal structural models to estimate the baseline risk of metastasis and the effect of ST (radiation therapy with or without hormone therapy) while accounting for selection into ST on the basis of PSA growth. The estimated model predicts the harm-benefit tradeoffs of ST within patient subgroups. The benefit of ST is the absolute reduction in the risk of metastasis within 10 years; the harm is the frequency of cancers that would not have metastasized in the patient's lifetime in the absence of ST (overtreatment). The adjusted hazard ratio associated with ST was 0.41 (95% CI, 0.31 to 0.55). Providing ST to all men at PSA-R reduced the risk of metastasis from 43% to 23% but led to 31% of men being overtreated (harm/benefit = 31/(43-23) = 1.6). Providing ST to men with Gleason score >7 reduced the risk of metastasis from 67% to 39%, with 13% of men being overtreated (harm/benefit = 13/(67-39) = 0.5). A quantitative framework that evaluates primary harms and benefits of ST after PSA-R will facilitate informed decision making. Immediate ST may be more appropriate in patient subgroups at elevated risk of metastasis.


TRIM68 regulates ligand-dependent transcription of androgen receptor in prostate cancer cells.
The androgen receptor (AR) is a transcription factor belonging to the family of nuclear receptors that mediate the action of androgen. AR plays an important role in normal development of the prostate, as well as in the progression of prostate cancer. AR is regulated by several posttranslational modifications, including phosphorylation, acetylation, and ubiquitination. In this study, we found that the putative E3 ubiquitin ligase TRIM68, which is preferentially expressed in prostate cancer cells, interacts with AR and enhances transcriptional activity of the AR in the presence of dihydrotestosterone. We also found that TRIM68 functionally interacts with TIP60 and p300, which act as coactivators of AR, and synergizes in the transactivation of AR. Overexpression of TRIM68 in prostate cancer cells caused an increase in secretion of prostate-specific antigen (PSA), one of the most reliable diagnostic markers for prostate cancer, whereas knockdown of TRIM68 attenuated the secretion of PSA and inhibited cell growth and colony-forming ability. Moreover, we showed that TRIM68 expression is significantly up-regulated in human prostate cancers compared with the expression in adjacent normal tissues. These results indicate that TRIM68 functions as a cofactor for AR-mediated transcription and is likely to be a novel diagnostic tool and a potentially therapeutic target for prostate cancer.


Breastfeeding, pregnant, and non-breastfeeding nor pregnant women's food consumption: A matched within-household analysis in India.
Promoting breastfeeding is major maternal and child health goal in India. It is unclear whether mothers receive additional food needed to support healthy breastfeeding. Using the latest National Family and Health Survey (2005-2006), we applied multilevel linear regression models to document correlates of nutrition for (n = 20,764) breastfeeding women. We then compared consumption of pulses, eggs, meat, fish, dairy, fruit, and vegetables across a sample of breastfeeding, non-breastfeeding/pregnant (NBP), and pregnant women (n = 3,409) matched within households and five-year age bands. We tested whether breastfeeding women had greater advantages in the 18 high-focus states of India's National Rural Health Mission (NRHM). Vegetarianism, caste, and religion were the strongest predictors of breastfeeding women's nutrition. Breastfeeding women had no nutritional advantage compared to NBP women, and were disadvantaged in their consumption of milk (b = -0.14) in low-focus states. Pregnant women were similarly disadvantaged in their consumption of milk in low-focus states (b = -0.32), but consumed vegetables more frequently (b = 0.12) than NBP women in high-focus states. Breastfeeding women do not receive nutritional advantages compared to NBP women. Targeted effort is needed to assess and improve nutritional adequacy for breastfeeding Indian women.


Salivary fluoride levels following application of fluoride varnish or fluoride rinse.
This study examined the concentration of fluoride in whole saliva over time following the application of a fluoride varnish or a single rinse with a fluoride solution. A two-period, two-treatment randomized cross-over experimental trial with a 2-week washout period was used with 16 adult subjects. In the first period, eight subjects rinsed once with a 0.05% NaF solution and 8 subjects had 5.0% NaF varnish applied to facial and lingual surfaces of 20 teeth. Stimulated whole saliva was collected at baseline, 5 and 15 min, 1, 2, 4, 8, 12, 24, 32, 48, 56, 72, 80, 96, 104 h. After the washout period each subject was switched to the other treatment and saliva was collected at the same intervals. Salivary fluoride content was measured with the micro-diffusion method. The NaF levels peaked at 5 min after application for both varnish (mean +/- SE 24.5 +/- 5.0 ppm) and rinse (3.2 +/- 0.8 ppm). Mean NaF levels returned to baseline, on average, within 2 h for the rinse and within 24 h for the varnish. The maximum fluoride levels were significantly greater (P < 0.01) with the varnish than with the rinse and remained above baseline levels for a longer duration. Salivary fluoride levels with the rinse returned to baseline, on average, in 2 h while they remained elevated for, on average, 24 h with the varnish. Salivary fluoride levels from the varnish were found to be comparable with those in previous studies for 1.1% neutral NaF.


Resistance to juvenile hormone and an insect growth regulator in Drosophila is associated with an altered cytosolic juvenile hormone-binding protein.
The Met mutant of Drosophila melanogaster is highly resistant to juvenile hormone III (JH III) or its chemical analog, methoprene, an insect growth regulator. Five major mechanisms of insecticide resistance were examined in Met and susceptible Met+ flies. These two strains showed only minor differences when penetration, excretion, tissue sequestration, or metabolism of [3H]JH III was measured. In contrast, when we examined JH III binding by a cytosolic binding protein from a JH target tissue, Met strains had a 10-fold lower binding affinity than did Met+ strains. Studies using deficiency-bearing chromosomes provide strong evidence that the Met locus controls the binding protein characteristics and may encode the protein. These studies indicate that resistance in Met flies results from reduced binding affinity of a cytosolic binding protein for JH III.


CYP51: A major drug target in the cytochrome P450 superfamily.
The cytochrome P540 (CYP) superfamily currently includes about 9000 proteins forming more than 800 families. The enzymes catalyze monooxygenation of a vast array of compounds and play essentially two roles. They provide biodefense (detoxification of xenobiotics, antibiotic production) and participate in biosynthesis of important endogenous molecules, particularly steroids. Based on these two roles, sterol 14/*alpha*/-demethylases (CYP51) belong to the second group of P450s. The CYP51 family, however, is very special as its members preserve strict functional conservation in enzyme activity in all biological kingdoms. At amino acid identity across the kingdoms as low as 25-30%, they all catalyze essentially the same three-step reaction of oxidative removal of the 14/*alpha*/-methyl group from the lanostane frame. This reaction is the required step in sterol biosynthesis of pathogenic microbes. We have shown that specific inhibition of protozoan CYP51 can potentially provide treatment for human trypanosomiases. Three sets of CYP51 inhibitors tested in vitro and in trypanosomal cells in this study include azoles [best results being 50% cell growth inhibition at <1 and at 1.3 muM for Trypanosoma cruzi (TC) and Trypanosoma brucei (TB), respectively], non-azole compounds (50% TC cell growth inhibition at 5 microM) and substrate analogs of the 14/*alpha*/-demethylase reaction. 32-Methylene cyclopropyl lanost-7-enol exhibited selectivity toward TC with 50% cell growth inhibition at 3 microM.


A family of Rhomboid intramembrane proteases activates all Drosophila membrane-tethered EGF ligands.
Drosophila has three membrane-tethered epidermal growth factor (EGF)-like proteins: Spitz, Gurken and Keren. Spitz and Gurken have been genetically confirmed to activate the EGF receptor, but Keren is uncharacterized. Spitz is activated by regulated intracellular translocation and cleavage by the transmembrane proteins Star and the protease Rhomboid-1, respectively. Rhomboid-1 is a member of a family of seven similar proteins in Drosophila. We have analysed four of these: all are proteases that can cleave Spitz, Gurken and Keren, and all activate only EGF receptor signalling in vivo. Star acts as an endoplasmic reticulum (ER) export factor for all three. The importance of this translocation is highlighted by the fact that when Spitz is cleaved by Rhomboids in the ER it cannot be secreted. Keren activates the EGF receptor in vivo, providing strong evidence that it is a true ligand. Our data demonstrate that all membrane-tethered EGF ligands in Drosophila are activated by the same strategy of cleavage by Rhomboids, which are ancient and widespread intramembrane proteases. This is distinct from the metalloprotease-induced activation of mammalian EGF-like ligands.


Developmental change in regional brain structure over 7 months in early adolescence: comparison of approaches for longitudinal atlas-based parcellation.
Early adolescence is a time of rapid change in neuroanatomy and sexual development. Precision in tracking changes in brain morphology with structural MRI requires image segmentation with minimal error. Here, we compared two approaches to achieve segmentation by image registration with an atlas to quantify regional brain structural development over a 7-month interval in normal, early adolescent boys and girls. Adolescents were scanned twice (average interval=7.3 months), yielding adequate data for analysis in 16 boys (baseline age 10.9 to 13.9 years; Tanner Stage=1 to 4) and 12 girls (baseline age=11.2 to 13.7 years; Tanner Stage=3 to 4). Brain volumes were derived from T1-weighted (SPGR) images and dual-echo Fast Spin-Echo (FSE) images collected on a GE 3T scanner with an 8-channel phased-array head coil and analyzed by registration-based parcellation using the SRI24 atlas. The "independent" method required two inter-subject registrations: both baseline (MRI 1) to atlas and follow-up (MRI 2) to the atlas. The "sequential" method required one inter-subject registration, which was MRI 1 to the atlas, and one intra-subject registration, which was MRI 2 to MRI 1. Gray matter/white matter/CSF were segmented in both MRI-1 and MRI-2 using FSL FAST with tissue priors also based on the SRI24 atlas. Gray matter volumes were derived for 10 cortical regions, gray+white matter volumes for 5 subcortical structures, and CSF volumes for 4 ventricular regions and the cortical sulci. Across the 15 tissue regions, the coefficient of variation (CV) of change scores across individuals was significantly lower for the sequential method (CV=3.02), requiring only one inter-subject registration, than for the independent method (CV=9.43), requiring two inter-subject registrations. Volume change based on the sequential method revealed that total supratentorial and CSF volumes increased, while cortical gray matter volumes declined significantly (p<0.01) in anterior (lateral and medial frontal, anterior cingulate, precuneus, and parietal) but not posterior (occipital, calcarine) cortical regions. These volume changes occurred in all boys and girls who advanced a step in Tanner staging. Subcortical structures did not show consistent changes. Thus, longitudinal MRI assessment using robust registration methods is sufficiently sensitive to identify significant regional brain changes over a 7-month interval in boys and girls in early adolescence. Increasing the temporal resolution of the retest interval in longitudinal developmental studies could increase accuracy in timing of peak growth of regional brain tissue and refine our understanding of the neural mechanisms underlying the dynamic changes in brain structure throughout adolescence.


Phase I clinical trial of curcumin, a chemopreventive agent, in patients with high-risk or pre-malignant lesions.
Curcumin (diferuloylmethane), a yellow substance from the root of the plant Curcuma longa Linn., has been demonstrated to inhibit carcinogenesis of murine skin, stomach, intestine and liver. However, the toxicology, pharmacokinetics and biologically effective dose of curcumin in humans have not been reported. This prospective phase-I study evaluated these issues of curcumin in patients with one of the following five high-risk conditions: 1) recently resected urinary bladder cancer; 2) arsenic Bowen's disease of the skin; 3) uterine cervical intraepithelial neoplasm (CIN); 4) oral leucoplakia; and 5) intestinal metaplasia of the stomach. Curcumin was taken orally for 3 months. Biopsy of the lesion sites was done immediately before and 3 months after starting curcumin treament. The starting dose was 500 mg/day. If no toxicity > or = grade II was noted in at least 3 successive patients, the dose was then escalated to another level in the order of 1,000, 2,000, 4,000, 8,000, and 12,000 mg/day. The concentration of curcumin in serum and urine was determined by high pressure liquid chromatography (HPLC). A total of 25 patients were enrolled in this study. There was no treatment-related toxicity up to 8,000 mg/day. Beyond 8,000 mg/day, the bulky volume of the drug was unacceptable to the patients. The serum concentration of curcumin usually peaked at 1 to 2 hours after oral intake of crucumin and gradually declined within 12 hours. The average peak serum concentrations after taking 4,000 mg, 6,000 mg and 8,000 mg of curcumin were 0.51 +/- 0.11 microM, 0.63 +/- 0.06 microM and 1.77 +/- 1.87 microM, respectively. Urinary excretion of curcumin was undetectable. One of 4 patients with CIN and 1 of 7 patients with oral leucoplakia proceeded to develop frank malignancies in spite of curcumin treatment. In contrast, histologic improvement of precancerous lesions was seen in 1 out of 2 patients with recently resected bladder cancer, 2 out of 7 patients of oral leucoplakia, 1 out of 6 patients of intestinal metaplasia of the stomach, I out of 4 patients with CIN and 2 out of 6 patients with Bowen's disease. In conclusion, this study demonstrated that curcumin is not toxic to humans up to 8,000 mg/day when taken by mouth for 3 months. Our results also suggest a biologic effect of curcumin in the chemoprevention of cancer.


[Psychedelics in the treatment of substance use disorders and psychosis].
After psychedelics were banned in 1968, the flourishing research on the use of psychedelics in patients with a mental disorder stopped abruptly. Recently, we see a renaissance of this research.
 AIM: To present an overview of what is known about the treatment of addiction and psychosis with psychedelics.
 METHOD: Literature study based on Medline en PubMed publications till December 2019.
 RESULTS: Studies on the effectiveness of psychedelics in the treatment of addiction and psychosis is still very limited in size and methodological quality. Nevertheless, most studies show positive effects of both classical and atypical psychedelics in a variety of addictions on motivation, craving, reduced consumption, and abstinence often following a single dose and with long-lasting benefits (3-24 months). Use of ketamine in patients with a psychosis stabilized on an antipsychotic might reduce negative symptoms.
 CONCLUSION: Before psychedelics can be used in standard clinical practice for the treatment of patients with an addiction or a psychosis, larger and methodologically better studies are needed. The use of psychedelics also creates an opportunity to better understand the shared underlying pathology of many different mental disorders.


Double-pass measurements of the retinal-image quality with unequal entrance and exit pupil sizes and the reversibility of the eye's optical system.
We have used a modified double-pass apparatus with unequal entrance and exit pupil sizes to measure the optical transfer function in the human eye and have applied the technique to three different problems. First, we confirm that in the eye the double-pass spread function is the cross correlation of the input spread function with the output spread function [J. Opt. Soc. Am. A 12, 195 (1995)]. Consequently, when entrance and exit pupil sizes are equal, phase information is lost from the double-pass images. Second, we show that in double-pass measurements the eye behaves like a reversible optical system. That is, when entrance and exit pupils are equal, the double-pass image results from two passes through an optical system having a transfer function that is the same in both directions. To test for reversibility in the living eye we have used a double-pass apparatus with different exit and entrance pupil sizes (one of them small enough to consider the eye diffraction limited), so that the ingoing and the outgoing transfer functions are different. The measured image quality was unchanged when the pupils were interchanged, i.e., when the first-pass entrance pupil size becomes the second-pass exit pupil size, and vice versa. Third, the technique provides a means for inferring the complete optical transfer function of the eye, including the phase transfer function, and the shape of the point-spread function.


Emergence of unique primate T-lymphotropic viruses among central African bushmeat hunters.
The human T-lymphotropic viruses (HTLVs) types 1 and 2 originated independently and are related to distinct lineages of simian T-lymphotropic viruses (STLV-1 and STLV-2, respectively). These facts, along with the finding that HTLV-1 diversity appears to have resulted from multiple cross-species transmissions of STLV-1, suggest that contact between humans and infected nonhuman primates (NHPs) may result in HTLV emergence. We investigated the diversity of HTLV among central Africans reporting contact with NHP blood and body fluids through hunting, butchering, and keeping primate pets. We show that this population is infected with a wide variety of HTLVs, including two previously unknown retroviruses: HTLV-4 is a member of a phylogenetic lineage that is distinct from all known HTLVs and STLVs; HTLV-3 falls within the phylogenetic diversity of STLV-3, a group not previously seen in humans. We also document human infection with multiple STLV-1-like viruses. These results demonstrate greater HTLV diversity than previously recognized and suggest that NHP exposure contributes to HTLV emergence. Our discovery of unique and divergent HTLVs has implications for HTLV diagnosis, blood screening, and potential disease development in infected persons. The findings also indicate that cross-species transmission is not the rate-limiting step in pandemic retrovirus emergence and suggest that it may be possible to predict and prevent disease emergence by surveillance of populations exposed to animal reservoirs and interventions to decrease risk factors, such as primate hunting.


Anterior universal spine system for adolescent idiopathic scoliosis: a follow-up study using scoliometer, real-time ultrasound and radiographs.
Nine patients with AIS treated surgically with anterior USS instrumentation were examined by several methods pre-operatively and at each of 8 weeks, 1 year and 2 years after surgery (mean age 14.6 years, girls 7, boys 2, thoracolumbar 7, lumbar 1, thoracic 1, left 7, right 2). The methods used were (1) Scoliometer to measure angle of trunk inclinations (ATIs) in the standing forward bending position at each of 10 levels and converted to 18 levels by a computer program, (2) real-time ultrasound in the prone position of laminal rotations at each of 1 8 levels from TI-SI, and (3) anteroposterior radiographs in the standing erect position measured for each of Cobb angle, segmental vertebral rotation (Perdriolle) and segmental vertebral translation from the Ti-Si line (horizontal translation of each vertebral centroid from the T1-S1 line). The findings were plotted graphically and segmentally for each of Scoliometer ATJs, ultrasound laminal rotations, and radiographic vertebral rotations and translations. Findings. Graphical representation of the data shows that the improvement brought about by surgery is most clearly and consistently evident for segmental vertebral translation. The statistical analysis shows that the radiological parameters (Cobb angle, apical vertebral rotation and apical vertebral translation) and ultrasound spinal (laminal) rotation do not change detectably in follow-up. The Scoliometer ATI findings show an increase from 4 degrees (at 8 weeks) to 7 degrees (at 2 years) which is statistically significant. The evidence from this small sample of patients is consistent with the view that the compared with posterior USS, anterior USS surgery for AIS results in (1) similar initial rib hump correction, and (2) less rib hump reassertion during follow-up. More data are needed to evaluate these views.


Omega-3 Fatty Acid Ingestion as a TBI Prophylactic.
Given the hazardous nature of combat operations and training exercises (e.g. airborne operations) conducted by the United States military, servicemembers are at high risk for sustaining a traumatic brain injury (TBI). Since the beginning of the Iraq and Afghanistan wars, almost a quarter of a million servicemembers have sustained a TBI.1 A large number of TBIs are a result of the concussive forces generated by improvised explosive devices (IED). A smaller number are a result of penetrating head wounds. Others may be caused by activities resulting in powerful acceleration, deceleration, or rotational forces. Therapies for treating TBI thus far have been limited. Much of the research conducted to date has focused on post-injury pharmacological interventions.2 Additionally, better protective equipment could help in preventing TBIs; however, these issues are outside the scope of this paper. A relatively new area of research is investigating prophylactic measures taken to lessen the effects of TBI. One such measure involves nutritional interventions and their effects on TBI severity. Therefore, the purpose of this paper is to elucidate the potential benefits of omega-3 fatty acid intake as it relates to TBI severity.


The effect of ovarian hormones on antioxidant enzyme activities in the brain of male rats.
The brain is widely responsive to gonadal hormones. The functional significance of ovarian hormones in the brain is evident from biochemical studies indicating that estradiol or progesterone treatment of testectomized rats produces changes of antioxidant enzyme activities. The effect of estradiol benzoate (EB) and progesterone (P) in the control of antioxidant (AO) enzyme activities was studied in the brain of adult male Wistar rats. The activities of catalase (CAT), glutathione peroxidase (GSH-Px), glutathione-S-transferase (GST) and glutathione reductase (GR) were measured in appropriate subcellular fractions, prepared from brains of animals belonging to various experimental groups. These groups were designed with the intention to follow changes in enzyme activities 2 h or 24 h after systemic administration of 5 microg EB or 2 mg P to testectomized (TX) animals. The obtained results show that both EB and P increase CAT activity, whereas EB decreases GSH-Px, GST and GR activities. These findings clearly show the modulatory role of EB and P in the control of enzymes responsible for the protection of rat nerve cells against oxidative damage caused by free oxygen radicals.


Targeting and killing of glioblastoma with activated T cells armed with bispecific antibodies.
Since most glioblastomas express both wild-type EGFR and EGFRvIII as well as HER2/neu, they are excellent targets for activated T cells (ATC) armed with bispecific antibodies (BiAbs) that target EGFR and HER2. ATC were generated from PBMC activated for 14 days with anti-CD3 monoclonal antibody in the presence of interleukin-2 and armed with chemically heteroconjugated anti-CD3 × anti-HER2/neu (HER2Bi) and/or anti-CD3 × anti-EGFR (EGFRBi). HER2Bi- and/or EGFRBi-armed ATC were examined for in vitro cytotoxicity using MTT and 51Cr-release assays against malignant glioma lines (U87MG, U118MG, and U251MG) and primary glioblastoma lines. EGFRBi-armed ATC killed up to 85% of U87, U118, and U251 targets at effector:target ratios (E:T) ranging from 1:1 to 25:1. Engagement of tumor by EGFRBi-armed ATC induced Th1 and Th2 cytokine secretion by armed ATC. HER2Bi-armed ATC exhibited comparable cytotoxicity against U118 and U251, but did not kill HER2-negative U87 cells. HER2Bi- or EGFRBi-armed ATC exhibited 50--80% cytotoxicity against four primary glioblastoma lines as well as a temozolomide (TMZ)-resistant variant of U251. Both CD133- and CD133+ subpopulations were killed by armed ATC. Targeting both HER2Bi and EGFRBi simultaneously showed enhanced efficacy than arming with a single BiAb. Armed ATC maintained effectiveness after irradiation and in the presence of TMZ at a therapeutic concentration and were capable of killing multiple targets. High-grade gliomas are suitable for specific targeting by armed ATC. These data, together with additional animal studies, may provide the preclinical support for the use of armed ATC as a valuable addition to current treatment regimens.


Air pollution from truck traffic and lung function in children living near motorways.
The contribution of motorized traffic to air pollution is widely recognized, but relatively few studies have looked at the respiratory health status of subjects living near busy roads. We studied children in six areas located near major motorways in the Netherlands. We measured lung function in the children, and we assessed their exposure to traffic-related air pollution using separate traffic counts for automobiles and trucks. We also measured air pollution in the children's schools. Lung function was associated with truck traffic density but had a lesser association with automobile traffic density. The association was stronger in children living closest (< 300 m) to the motorways. Lung function was also associated with the concentration of black smoke, measured inside the schools, as a proxy for diesel exhaust particles. The associations were stronger in girls than in boys. The results indicate that exposure to traffic-related air pollution, in particular diesel exhaust particles, may lead to reduced lung function in children living near major motorways.


Intracellular survival of Burkholderia pseudomallei.
Burkholderia pseudomallei is the causative agent of melioidosis, a disease being increasingly recognized as an important cause of morbidity and mortality in many regions of the world. Several features of melioidosis suggest that B. pseudomallei is a facultative intracellular pathogen. This study was designed to assess the ability of B. pseudomallei to invade and survive in eukaryotic cells. We have shown that B. pseudomallei has the capacity to invade cultured cell lines, including HeLa, CHO, A549, and Vero cells. We have demonstrated intracellular survival of B. pseudomallei in professional phagocytic cells, including rat alveolar macrophages. B pseudomallei was localized inside vacuoles in human monocyte-like U937 cells, a histiocytic lymphoma cell line with phagocytic properties. Additionally, electron microscopic visualization of B. pseudomallei-infected HeLa cells and polymorphonuclear leukocytes confirmed the presence of intracellular bacteria within membrane-bound vacuoles. B. pseudomallei was found to be resistant to the cationic peptide protamine and to purified human defensin HNP-1.


The role of gammadelta T cells in induction of bacterial antigen-specific protective CD8+ cytotoxic T cells in immune response against the intracellular bacteria Listeria monocytogenes.
The role of T-cell receptor (TCR) gammadelta T cells in the induction of protective TCR alphabeta T cells against infection by the intracellular bacteria Listeria monocytogenes was analysed. We found that depletion of gammadelta T cells by anti-TCR delta monoclonal antibody treatment before intravenous immunization of mice with a sublethal dose of viable L. monocytogenes resulted in reduction of protection against secondary challenge infection in the immunized mice. The gammadelta T-cell depletion also reduced induction of protective alphabeta T cells capable of transferring the protection against challenge infection of L. monocytogenes into naive mice. Furthermore, the protective T cells that were affected by the gammadelta T-cell depletion were suggested to be CD8+ cytotoxic T cells rather than CD4+ T cells by the following observations. First, induction of cytotoxic T lymphocytes specific to a L. monocytogenes-derived H-2Kd-restricted peptide (listeriolysin O 91-99) was significantly suppressed by gammadelta T-cell depletion before immunization. Second, gammadelta T-cell depletion did not affect cytokine production and proliferation of T cells from immunized mice in response to in vitro stimulation with heat-killed Listeria which preferentially stimulates CD4+ T cells. Third, CD8+ alphabeta T cells from control immunized mice transferred protection against infection of L. monocytogenes into naive mice but only a limited degree of protection was transferred by CD8+ T cells from the gammadelta T-cell-depleted immunized mice; and fourth, CD4+ alphabeta T cells from the gammadelta T-cell-depleted mice transferred a similar level of protection as those from the control immunized mice. All these results suggest that gammadelta T cells participate in establishment of protective immunity against intracellular bacteria by supporting priming of bacterial antigen-specific CD8+ cytotoxic T cells.


Identification of candidate genes for controlling development of the basilar pons by differential display PCR.
The basilar pons, a major hindbrain nucleus involved in sensory-motor integration, has become a model system for studying long-distance neuronal migration, axon-target recognition by collateral branching, and the formation of patterned axonal projections. To identify genes potentially involved in these developmental events, we have performed a differential display PCR screen comparing RNA isolated from the developing basilar pons with RNA obtained from developing cerebellum and olfactory bulb, as well as the mature basilar pons. Using 400 different combinations of primers, we screened more than 11,000 labeled DNA fragments and identified 201 that exhibited higher expression in the basilar pons than in the control tissues. From these, 138 distinct gene fragments were cloned. The differential expression of a large subset of these fragments was confirmed using RNase protection assays. In situ hybridization analysis revealed that the expression of many of these genes is limited to the basilar pons and only a few other brain regions, suggesting that they may play specific roles in pontine development.


Serum markers of liver fibrosis: combining the BIPED classification and the neo-epitope approach in the development of new biomarkers.
Fibrosis is a central histological feature of chronic liver diseases and is characterized by the accumulation and reorganization of the extracellular matrix. The gold standard for assessment of fibrosis is histological evaluation of a percutaneous liver biopsy. Albeit a considerable effort have been invested in finding alternative non-invasive approaches, these have not been sufficiently successful to replace biopsy assessment. To identify the extracellular matrix proteins of interest, that as protein degradation fragments produced during extracellular matrix metabolism neo-epitopes, may be targeted for novel biochemical marker development in fibrosis. We used the recently proposed BIPED system (Burden of disease, Investigative, Prognostic, Efficacy and Diagnostic) to characterise present serological markers. Pubmed was search for keywords; Liver fibrosis, neo-epitopes, biomarkers, clinical trail, extra cellular matrix, protease, degradation, fragment. Implementation of BIPED categorization in the development and validation of fibrosis biomarkers to simplify and standardize the use of existing and future biomarkers seems advantageous. In addition, a systematic use of the neo-epitope approach, i.e. the quantification of peptide epitopes generated from enzymatic cleavage of proteins during extracellular remodeling, may prove productive in the quest to find new markers of liver fibrosis.


High Incidence of Invasive Group A Streptococcus Disease Caused by Strains of Uncommon emm Types in Thunder Bay, Ontario, Canada.
An outbreak of type emm59 invasive group A Streptococcus (iGAS) disease was declared in 2008 in Thunder Bay District, Northwestern Ontario, 2 years after a countrywide emm59 epidemic was recognized in Canada. Despite a declining number of emm59 infections since 2010, numerous cases of iGAS disease continue to be reported in the area. We collected clinical information on all iGAS cases recorded in Thunder Bay District from 2008 to 2013. We also emm typed and sequenced the genomes of all available strains isolated from 2011 to 2013 from iGAS infections and from severe cases of soft tissue infections. We used whole-genome sequencing data to investigate the population structure of GAS strains of the most frequently isolated emm types. We report an increased incidence of iGAS in Thunder Bay compared to the metropolitan area of Toronto/Peel and the province of Ontario. Illicit drug use, alcohol abuse, homelessness, and hepatitis C infection were underlying diseases or conditions that might have predisposed patients to iGAS disease. Most cases were caused by clonal strains of skin or generalist emm types (i.e., emm82, emm87, emm101, emm4, emm83, and emm114) uncommonly seen in other areas of the province. We observed rapid waxing and waning of emm types causing disease and their replacement by other emm types associated with the same tissue tropisms. Thus, iGAS disease in Thunder Bay District predominantly affects a select population of disadvantaged persons and is caused by clonally related strains of a few skin and generalist emm types less commonly associated with iGAS in other areas of Ontario.


Combined HDAC and BET Inhibition Enhances Melanoma Vaccine Immunogenicity and Efficacy.
The combined inhibition of histone deacetylases (HDAC) and the proteins of the bromodomain and extraterminal (BET) family have recently shown therapeutic efficacy against melanoma, pancreatic ductal adenocarcinoma, testicular, and lymphoma cancers in murine studies. However, in such studies, the role of the immune system in therapeutically controlling these cancers has not been explored. We sought to investigate the effect of the HDAC inhibitor romidepsin (RMD) and the BET inhibitor IBET151, both singly and in combination, on vaccine-elicited immune responses. C57BL/6 mice were immunized with differing vaccine systems (adenoviral, protein) in prime-boost regimens under treatment with RMD, IBET151, or RMD+IBET151. The combined administration of RMD+IBET151 during vaccination resulted in a significant increase in the frequency and number of Ag-specific CD8+ T cells. RMD+IBET151 treatment significantly increased the frequency of vaccine-elicited IFN-γ+ splenic CD8+ T cells and conferred superior therapeutic and prophylactic protection against B16-OVA melanoma. RNA sequencing analyses revealed strong transcriptional similarity between RMD+IBET151 and untreated Ag-specific CD8+ T cells except in apoptosis and IL-6 signaling-related genes that were differentially expressed. Serum IL-6 was significantly increased in vivo following RMD+IBET151 treatment, with recombinant IL-6 administration replicating the effect of RMD+IBET151 treatment on vaccine-elicited CD8+ T cell responses. IL-6 sufficiency for protection was not assessed. Combined HDAC and BET inhibition resulted in greater vaccine-elicited CD8+ T cell responses and enhanced therapeutic and prophylactic protection against B16-OVA melanoma. Increased IL-6 production and the differential expression of pro- and anti-apoptotic genes following RMD+IBET151 treatment are likely contributors to the enhanced cancer vaccine responses.


Isolation and characterization of a sea urchin zygote cortex that supports in vitro contraction and reactivation of furrowing.
The isolation of the cortex of the sea urchin blastomere by detergent lysis was explored with the aim of analyzing components important in the structure and function of the cortical cytoskeleton, and their relationship to such phenomena as contraction. Buffered EGTA medium supplemented with isotonic glycerol and with magnesium, at a level close to the reported internal cellular concentration, yields stable cytoskeletal cortices that retain their spherical shape. Cortices prepared this way contain actin, myosin, fascin and spectrin, components normally associated with the cortical cytoskeleton in a similar distribution to that in intact zygotes. They retain the organized cortical filamentous structure, including the actin-fascin bundles that form cores of microvilli. ATP and NaCl caused changes in cortical shape, described as either contraction or expansion, respectively. Spectrin, but not myosin, was partially extracted by NaCl, resulting in expansion of the cortex that suggests a role for spectrin in maintenance of cortical structure. ATP (but not ADP nor ATP gamma S), which caused the partial removal of myosin and spectrin, led to the contraction of the cortex, consistent with a role for myosin in cortical tension. In cortices isolated from dividing eggs, the zygotes retained their cleavage furrows and ATP induced continuation of furrow progression. This preparation appears to be a useful in vitro model for cytokinesis.


Radiation Emergency Medical Preparedness in Japan: A Survey of Nuclear Emergency Core Hospitals.
Based on experiences following the Great East Japan Earthquake and nuclear power plant accident in 2011, Nuclear Emergency Core Hospitals (NECHs) were designated as centers for radiation disaster management in Japan. This study aimed to investigate their current status and identify areas for improvement. This cross-sectional study was conducted in October 2018. Demographic data were collected by a questionnaire with free text responses about attitudes toward NECHs. Considerations regarding risk communications during a radiation disaster were analyzed using qualitative text mining analysis. A total of 36 hospitals participated in this study. Only 31% of NECHs anticipated a radiation disaster. The importance of business continuity plans and risk communications was shown. Text analysis identified 7 important categories for health care workers during a radiation disaster, including media response, communications to hospital staff, risk communications, radiation effects on children, planning for a radiation disaster in the region, rumors, and the role in the region. The radiation disaster medical system and NECHs in Japan were surveyed. The importance of risk communications, planning for a radiation disaster in each region, and the role in the region are identified as issues that need to be addressed.


Serotonin and emotional processing: does it help explain antidepressant drug action?
There is growing interest in the effects of antidepressant drug treatment on measures of emotional processing. Such actions may help us understand the role of monoamines in emotional dysfunction in depression and how antidepressant drug treatments work. Recent studies suggest that decreasing central serotonin function with tryptophan depletion can reinstate negative biases in recovered depressed patients, even at doses insufficient to induce changes in mood. Conversely, antidepressant drug administration increases the processing of positive emotional information in healthy volunteers and acutely depressed patients early in treatment. This increase in positive bias may provide a platform for subsequent cognitive restructuring and learning which contributes to the evolution of symptom change in depression. Functional neuroimaging studies suggest that these early antidepressant effects involve fronto-limbic and extra-striate circuitry suggestive of actions on both the initial appraisal and attentional processing of affective stimuli. This approach may therefore provide a framework for linking psychological and biological processes in emotional disorders and their treatment. Antidepressants may not directly modulate mood and anxiety but rather allow a different perspective for our ongoing evaluation of our self, the world and the future.


Moving targets: Promoting physical activity in public spaces via open streets in the US.
Popularity of Open Streets, temporarily opening streets to communities and closing streets to vehicles, in the US has recently surged. As of January 2016, 122 cities have hosted an Open Streets program. Even with this great expansion, the sustainability of Open Streets remains a challenge in many cities and overall Open Streets in the US differ from their successful counterparts in Central and South America. Between summer 2015 and winter 2016, we reviewed the websites and social media of the 122 identified programs and interviewed 32 unique Open Streets programs. Websites and social media were reviewed for program initiation, number of Open Streets days, length of routes, duration of program, and reported participation. Interview questions focused on barriers and facilitators of expanding Open Streets and specific questioning regarding local evaluation activities. All interviews were transcribed verbatim and analyzed with constant comparative methodology. Over three-quarters of US Open Streets programs have been initiated since 2010, with median frequency of one time per year, 4h per date, and 5000-9999 participants. Seventy-seven percent of program routes are under 5km in length. Success of programs was measured by enthusiasm, attendance, social media, survey metrics, and sustainability. Thirteen of 32 program organizers expressed interest in expanding their programs to 12 dates per year, but noted consistent barriers to expansion including funding, permitting, and branding. Though many cities now host Open Streets programs, their ability to effect public health remains limited with few program dates per year. Coordinated efforts, especially around funding, permitting, and branding may assist in expanding program dates.