Mapping QTLs and association of differentially expressed gene transcripts for multiple agronomic traits under different nitrogen levels in sorghum.
Sorghum is an important C4 crop which relies on applied Nitrogen fertilizers (N) for optimal yields, of which substantial amounts are lost into the atmosphere. Understanding the genetic variation of sorghum in response to limited nitrogen supply is important for elucidating the underlying genetic mechanisms of nitrogen utilization. A bi-parental mapping population consisting of 131 recombinant inbred lines (RILs) was used to map quantitative trait loci (QTLs) influencing different agronomic traits evaluated under normal N (100 kg.ha(-1) fertilizer) and low N (0 kg.ha(-1) fertilizer) conditions. A linkage map spanning 1614 cM was developed using 642 polymorphic single nucleotide polymorphisms (SNPs) detected in the population using Genotyping-By-Sequencing (GBS) technology. Composite interval mapping detected a total of 38 QTLs for 11 agronomic traits tested under different nitrogen levels. The phenotypic variation explained by individual QTL ranged from 6.2 to 50.8%. Illumina RNA sequencing data generated on seedling root tissues revealed 726 differentially expressed gene (DEG) transcripts between parents, of which 108 were mapped close to the QTL regions. Co-localized regions affecting multiple traits were detected on chromosomes 1, 5, 6, 7 and 9. These potentially pleiotropic regions were coincident with the genomic regions of cloned QTLs, including genes associated with flowering time, Ma3 on chromosome 1 and Ma1 on chromosome 6, gene associated with plant height, Dw2 on chromosome 6. In these regions, RNA sequencing data showed differential expression of transcripts related to nitrogen metabolism (Ferredoxin-nitrate reductase), glycolysis (Phosphofructo-2-kinase), seed storage proteins, plant hormone metabolism and membrane transport. The differentially expressed transcripts underlying the pleiotropic QTL regions could be potential targets for improving sorghum performance under limited N fertilizer through marker assisted selection.


Ongoing mumps outbreak in Israel, January to August 2017.
In Israel, 262 mumps cases were registered between 1 January and 28 August 2017 despite a vaccine coverage of ≥ 96%. The majority (56.5%) of cases were adolescents and young adults between 10 and 24 years of age. Nearly twice as many cases were reported in males than in females. Sequence information identified genotype G and suggested specific transmission chains in different religious communities, with the Muslim population in Jerusalem being most severely affected.


A serpin that regulates immune melanization in the respiratory system of Drosophila.
Epithelial tissues facing the external environment are essential to combating microbial infection. In addition to providing a physical barrier, epithelial tissues mount chemical defenses to prevent invasion of internal tissues by pathogens. Here, we describe that the melanization reaction implicated in host defense is activated in the respiratory system, the trachea, of Drosophila. Tracheal melanization can be activated by the presence of microorganisms but is normally blocked by Spn77Ba, a protease inhibitor in the serpin family. Spn77Ba inhibits a protease cascade involving the MP1 and MP2 proteases that activates phenol oxidase, a key enzyme in melanin biosynthesis. Unexpectedly, we found that tracheal melanization resulting from Spn77Ba disruption induces systemic expression of the antifungal peptide Drosomycin via the Toll pathway. Such signaling between local and systemic immune responses could represent an alarm mechanism that prepares the host in case a pathogen breaches epithelial defenses to invade internal tissues.


Quantitative morphometric analysis of hepatocellular carcinoma: development of a programmed algorithm and preliminary application.
The quantitative relationship between tumor morphology and malignant potential has not been explored in liver tumors. We designed a computer algorithm to analyze shape features of hepatocellular carcinoma (HCC) and tested feasibility of morphologic analysis. Cross-sectional images from 118 patients diagnosed with HCC between 2007 and 2010 were extracted at the widest index tumor diameter. The tumor margins were outlined, and point coordinates were input into a MATLAB (MathWorks Inc., Natick, Massachusetts, USA) algorithm. Twelve shape descriptors were calculated per tumor: the compactness, the mean radial distance (MRD), the RD standard deviation (RDSD), the RD area ratio (RDAR), the zero crossings, entropy, the mean Feret diameter (MFD), the Feret ratio, the convex hull area (CHA) and perimeter (CHP) ratios, the elliptic compactness (EC), and the elliptic irregularity (EI). The parameters were correlated with the levels of alpha-fetoprotein (AFP) as an indicator of tumor aggressiveness. The quantitative morphometric analysis was technically successful in all cases. The mean parameters were as follows: compactness 0.88±0.086, MRD 0.83±0.056, RDSD 0.087±0.037, RDAR 0.045±0.023, zero crossings 6±2.2, entropy 1.43±0.16, MFD 4.40±3.14 cm, Feret ratio 0.78±0.089, CHA 0.98±0.027, CHP 0.98±0.030, EC 0.95±0.043, and EI 0.95±0.023. MFD and RDAR provided the widest value range for the best shape discrimination. The larger tumors were less compact, more concave, and less ellipsoid than the smaller tumors (P < 0.0001). AFP-producing tumors displayed greater morphologic irregularity based on several parameters, including compactness, MRD, RDSD, RDAR, entropy, and EI (P < 0.05 for all). Computerized HCC image analysis using shape descriptors is technically feasible. Aggressively growing tumors have wider diameters and more irregular margins. Future studies will determine further clinical applications for this morphologic analysis.


Opportunities to improve diagnosis in emergency transfers to the pediatric intensive care unit.
Late recognition of in-hospital deterioration is a source of preventable harm. Emergency transfers (ET), when hospitalized patients require intensive care unit (ICU) interventions within 1 h of ICU transfer, are a proximal measure of late recognition associated with increased mortality and length of stay (LOS). To apply diagnostic process improvement frameworks to identify missed opportunities for improvement in diagnosis (MOID) in ETs and evaluate their association with outcomes. A single-center retrospective cohort study of ETs, January 2015 to June 2019. ET criteria include intubation, vasopressor initiation, or ≥ $\ge \phantom{\rule{}{0ex}}$ 60 mL/kg fluid resuscitation 1 h before to 1 h after ICU transfer. The primary exposure was the presence of MOID, determined using SaferDx. Cases were screened by an ICU and non-ICU physician. Final determinations were made by an interdisciplinary group. Diagnostic process improvement opportunities were identified. Primary outcomes were in-hospital mortality and posttransfer LOS, analyzed by multivariable regression adjusting for age, service, deterioration category, and pretransfer LOS. MOID was identified in 37 of 129 ETs (29%, 95% confidence interval [CI] 21%-37%). Cases with MOID differed in originating service, but not demographically. Recognizing the urgency of an identified condition was the most common diagnostic process opportunity. ET cases with MOID had higher odds of mortality (odds ratio 5.5; 95% CI 1.5-20.6; p = .01) and longer posttransfer LOS (rate ratio 1.7; 95% CI 1.1-2.6; p = .02). MOID are common in ETs and are associated with increased mortality risk and posttransfer LOS. Diagnostic improvement strategies should be leveraged to support earlier recognition of clinical deterioration.


Ultrasonography incorrectly diagnoses gallbladder polyps.
To determine the accuracy of ultrasound-diagnosed polypoid lesions of the gallbladder in their institution, the authors reviewed the records of 41 patients with polypoid lesions of the gallbladder who underwent cholecystectomy, and collected data concerning age, sex, symptoms, and histopathologic diagnosis. Histopathologic evaluation confirmed polyps in only two patients (4.9%) categorized as having polypoid lesions of the gallbladder. Most specimens from patients with ultrasonography reports suggesting small polyps manifested cholesterolosis (17 of 41) or cholelithiasis (15 of 41). No specimen harbored malignancy. Mean patient age was 47.4 years, and the most common symptoms were pain (85%), nausea (44%), vomiting (29%), and abnormal liver function test results (14%). The accuracy of sonography for diagnosing polypoid lesions of the gallbladder was poor. Many of the small polyps seen on sonography most likely represented a stone embedded in the gallbladder wall or other abnormality. Because of the likelihood of cholelithiasis, the authors recommend that patients with biliary symptoms and ultrasonography findings suggesting polypoid lesions of the gallbladder undergo cholecystectomy.


Ultrasound Capsule Endoscopy With a Mechanically Scanning Micro-ultrasound: A Porcine Study.
Wireless capsule endoscopy has been used for the clinical examination of the gastrointestinal (GI) tract for two decades. However, most commercially available devices only utilise optical imaging to examine the GI wall surface. Using this sensing modality, pathology within the GI wall cannot be detected. Micro-ultrasound (μUS) using high-frequency (>20 MHz) ultrasound can provide a means of transmural or cross-sectional image of the GI tract. Depth of imaging is approximately 10 mm with a resolution of between 40-120 μm that is sufficient to differentiate between subsurface histologic layers of the various regions of the GI tract. Ultrasound capsule endoscopy (USCE) uses a capsule equipped with μUS transducers that are capable of imaging below the GI wall surface, offering thereby a complementary sensing technique to optical imaging capsule endoscopy. In this work, a USCE device integrated with a ∼30 MHz ultrasonic transducer was developed to capture a full 360° image of the lumen. The performance of the device was initially evaluated using a wire phantom, indicating an axial resolution of 69.0 μm and lateral resolution of 262.5 μm. Later, in vivo imaging performance was characterised in the oesophagus and small intestine of anaesthetized pigs. The reconstructed images demonstrate clear layer differentiation of the lumen wall. The tissue thicknesses measured from the B-scan images show good agreement with ex vivo images from the literature. The high-resolution ultrasound images in the in vivo porcine model achieved with this device is an encouraging preliminary step in the translation of these devices toward future clinical use.


Early life influences on serum 1,25 (OH) vitamin D.
There is increasing evidence to support the role of the intrauterine and early postnatal environment in determining adult bone mass and risk of fracture. The mechanisms by which this occurs are uncertain but include perturbations in several endocrine axes. Vitamin D is integrally involved in bone metabolism and is therefore an ideal candidate. This study assesses whether birthweight and weight at 1 year of age are associated with the calcium vitamin D axis in elderly women. Vitamin D metabolites, parathyroid hormone, bone mineral density and biochemical markers of bone turnover were measured in 129 healthy women (mean age 65.5 years) from the MRC Hertfordshire Cohort Study whose birthweight and weight at 1 year were available from records. Serum 1,25 (OH)2 vitamin D concentrations were reduced with increasing weight at 1 year (19.1% reduction between the lowest and highest tertiles, P <0.01). A similar, but weaker trend was seen for birthweight. These associations were not explained by serum levels of serum calcium, phosphate, PTH, creatinine or sex hormones. The association of serum calcium with 1,25 (OH)2 vitamin D was greatest in the lowest tertile with little association in the highest tertile suggesting an increased sensitivity of renal 1-alpha hydroxylase in the lowest tertile. Highest levels of 1,25 (OH)2 vitamin D were associated with low BMD and high levels of urinary N-telopeptide suggesting that vitamin D metabolism may mediate the intrauterine and early postnatal environmental effects on adult BMD.


Identifying and treating refractory ITP: difficulty in diagnosis and role of combination treatment.
Immune thrombocytopenia (ITP) is the most common acquired thrombocytopenia after chemotherapy-induced thrombocytopenia. Existing guidelines describe the management and treatment of most patients who, overall, do well, even if they present with chronic disease, and they are usually not at a high risk for bleeding; however, a small percentage of patients is refractory and difficult to manage. Patients classified as refractory have a diagnosis that is not really ITP or have disease that is difficult to manage. ITP is a diagnosis of exclusion; no specific tests exist to confirm the diagnosis. Response to treatment is the only affirmative confirmation of diagnosis. However, refractory patients do not respond to front-line or other treatments; thus, no confirmation of diagnosis exists. The first section of this review carefully evaluates the diagnostic considerations in patients with refractory ITP. The second section describes combination treatment for refractory cases of ITP. The reported combinations are divided into the era before thrombopoietin (TPO) and rituximab and the current era. Current therapy appears to have increased effectiveness. However, the definition of refractory, if it includes insufficient response to TPO agents, describes a group with more severe and difficult-to-treat disease. The biology of refractory ITP is largely unexplored and includes oligoclonality, lymphocyte pumps, and other possibilities. Newer treatments, especially rapamycin, fostamatinib, FcRn, and BTK inhibitors, may be useful components of future therapy given their mechanisms of action; however, TPO agents, notwithstanding failure as monotherapy, appear to be critical components. In summary, refractory ITP is a complicated entity in which a precise specific diagnosis is as important as the development of effective combination treatments.


Anatomical Study of the Extreme Lateral Transpsoas Lumbar Interbody Fusion with Application to Minimizing Injury to the Kidney.
Objective Since the extreme lateral lumbar interbody fusion procedure was first reported by Ozgur in 2006, a large number of clinical studies have been published. Anatomical studies which explore methods to avoid visceral structures, such as the kidney, with this approach have not been examined in detail. We dissected the retroperitoneal space to analyze how the extreme lateral transpsoas approach to the lumbar spine could damage the kidney and related structures. Methods Eight sides from four fresh Caucasian cadavers were used for this study. The latissimus dorsi muscle and the thoracolumbar fascia were dissected to open the retroperitoneum. The fat tissue was removed. Steel wires were then put into the intervertebral disc spaces. Finally, the closest distance between kidney and wires on each interdiscal space was measured. Results The closest distance from the wire in the interdiscal space on L1/2, L2/3 and L3/4 to the kidney ranged from 13.2 mm to 32.9 mm, 20.0 mm to 27.7 mm, and 20.5 mm to 46.6 mm, respectively. The distance from the kidney to the interdiscal space at L4/5 was too great to be considered applicable to this study. Conclusions The results of this study might help surgeons better recognize the proximity of the kidney and avoid injury to it during the extreme lateral transpsoas approach to the lumbar spine.


Earlier menarche is associated with fatty liver and abdominal ectopic fat in midlife, independent of young adult BMI: The CARDIA study.
The hypothesis that earlier menarche is associated with higher non alcoholic fatty liver disease (NAFLD) and ectopic adiposity, independent of young adult body mass index (BMI), was tested. The data from 1,214 black and white women in the Coronary Artery Risk Development in Young Adults (CARDIA) study who reliably reported menarche age at exam years 0 and 2, who had multiple-slice abdominal computed tomography (CT) at exam year 25, and who had no known liver disease or secondary causes of steatosis were included. Women were aged 18-30 at year 0 and 43-55 at year 25. Liver attenuation, visceral adipose tissue (VAT), subcutaneous adipose tissue (SAT), and inter-muscular adipose tissue (IMAT) were derived from CT. NAFLD was defined as liver attenuation less than 51 Hounsfield units. One-year earlier menarche was associated with higher NAFLD (RR = 1.15; 95% CI: 1.07, 1.24), and VAT (6.7 cc; 95% CI: 4.3, 9.0 cc), IMAT (1.0 cc; 95% CI: 0.6, 1.4 cc), and SAT (19.6 cc; 95% CI: 13.2, 26.0 cc) after confounder adjustment. Associations remained significant (P < 0.05) after further adjustment for year 0 BMI. Only VAT remained significant (P = 0.047) after adjustment for weight gain between years 0 and 25. Earlier menarche is positively associated with NAFLD and ectopic fat independent of confounders and young adult BMI. Weight gain between young adulthood and midlife explains some of this association.


The neural correlates of continuous feedback processing.
Feedback processing is commonly studied by analyzing the brain's response to discrete rather than continuous events. Such studies have led to the hypothesis that rapid phasic midbrain dopaminergic activity tracks reward prediction errors (RPEs), the effects of which are measurable at the scalp via electroencephalography (EEG). Although studies using continuous feedback are sparse, recent animal work suggests that moment-to-moment changes in reward are tracked by slowly ramping midbrain dopaminergic activity. Some have argued that these ramping signals index state values rather than RPEs. Our goal here was to develop an EEG measure of continuous feedback processing in humans, then test whether its behavior could be accounted for by the RPE hypothesis. Participants completed a stimulus-response learning task in which a continuous reward cue gradually increased or decreased over time. A regression-based unmixing approach revealed EEG activity with a topography and time course consistent with the stimulus-preceding negativity (SPN), a scalp potential previously linked to reward anticipation and tonic dopamine release. Importantly, this reward-related activity depended on outcome expectancy: as predicted by the RPE hypothesis, activity for expected reward cues was reduced compared to unexpected reward cues. These results demonstrate the possibility of using human scalp-recorded potentials to track continuous feedback processing, and test candidate hypotheses of this activity.


Measuring the symptom burden associated with the treatment of chronic myeloid leukemia.
We developed a module of the MD Anderson Symptom Inventory (MDASI) for patients with chronic myeloid leukemia (CML). To develop the MDASI-CML, we identified CML-specific symptoms from qualitative interviews with 35 patients. A list of candidate symptoms was reduced by a panel of patients, caregivers, and clinicians to the 13 core MDASI symptom items and 6 CML-specific items; these items were subsequently administered to 30 patients. Cognitive debriefing confirmed that the items were clear, relevant, and easy to use. One additional CML-specific symptom item was added, for a total of 7. The refined MDASI-CML was administered to 152 patients once every 2 weeks for 1 year. The content, concurrent, known-group, and construct validity of the MDASI-CML were evaluated. The internal consistency and test-retest reliabilities of the module were adequate. Longitudinal analysis showed relatively stable symptom severity scores over time. The most severe symptoms were fatigue, drowsiness, disturbed sleep, muscle soreness and cramping, and trouble remembering things. Approximately one-third of the patients who completed the MDASI-CML reported persistent moderate-to-severe symptoms. The MDASI-CML is a valid and reliable symptom assessment instrument that can be used in clinical studies of symptom status in patients with CML.


Are the King LTS Laryngeal Tube and the Ambu AuraOnce Laryngeal Mask Useful Airway Adjuncts for Military Medics?
Airway management is a critical skill that may be essential in the battlefield. The aim of this study was to determine if combat life savers and medics with no or limited clinical experience could provide airway control using the disposable laryngeal suction tube (LTS-D) and the Ambu® AuraOnce ™ disposable laryngeal mask (LMA). Eight military medics with limited clinical experience and no airway management experience secured the airway on 19 intrahospital anesthetized patients using the LTS-D and the LMA. Each patient was treated with both airway adjuncts in a randomized order. Each medic was studied on at least two and not more than three patients. Success of insertion, number of attempts, and time to correct placement with verified ventilation were recorded. When using the LTS-D, the first-attempt success rate for the medics with the first patient was 50% (four of eight) and 75% (six of eight) on the second attempt. With the second patient, the first-attempt success rate was 87.5% (seven of eight), and on the third patient, it was 100% (three of three). For all patients, the accumulated first-attempt success rate was 73.7% (14 of 19) and 84.2% (16 or 19) for second attempts with the LTS-D. The success rate on first attempt with the LMA was 100% (19 of 19) on the first patient. On the second intervention with the LMA, there was a significant decrease in mean time to verified ventilation, from 56.4 seconds to 27.7 seconds (ρ = .043), and time to verified ventilation with no leakage, from 61.3 seconds to 32.3 seconds (ρ = .029). Insertion attempts for three LTS-Ds and one LMA failed. This study suggests that for the safe use of the laryngeal tube suction by inexperienced airway providers, a clinical training program is required. The LMA proved superior to the LTS-D with a 100% success rate at the first attempt, which suggests that minimal training is needed before clinical use. We therefore recommend the LMA as the primary airway adjunct for military medics, despite a hypothetically inferior protection from aspiration.


Skin mucus of Cyprinus carpio inhibits cyprinid herpesvirus 3 binding to epidermal cells.
Cyprinid herpesvirus 3 (CyHV-3) is the aetiological agent of a mortal and highly contagious disease in common and koi carp. The skin is the major portal of entry of CyHV-3 in carp after immersion in water containing the virus. In the present study, we used in vivo bioluminescence imaging to investigate the effect of skin mucus removal and skin epidermis lesion on CyHV-3 entry. Physical treatments inducing removal of the mucus up to complete erosion of the epidermis were applied on a defined area of carp skin just before inoculation by immersion in infectious water. CyHV-3 entry in carp was drastically enhanced on the area of the skin where the mucus was removed with or without associated epidermal lesion. To investigate whether skin mucus inhibits CyHV-3 binding to epidermal cells, tail fins with an intact mucus layer or without mucus were inoculated ex vivo. While electron microscopy examination revealed numerous viral particles bound on the fins inoculated after mucus removal, no particle could be detected after infection of mucus-covered fins. Finally, anti-CyHV-3 neutralising activity of mucus extract was tested in vitro. Incubation of CyHV-3 with mucus extract reduced its infectivity in a dose dependent manner. The present study demonstrates that skin mucus removal and epidermal lesions enhance CyHV-3 entry in carp. It highlights the role of fish skin mucus as an innate immune protection against viral epidermal entry.


Photopolymerization of Dienoyl Lipids Creates Planar Supported Poly(lipid) Membranes with Retained Fluidity.
Polymerization of substrate-supported bilayers composed of dienoylphosphatidylcholine (PC) lipids is known to greatly enhance their chemical and mechanical stability; however, the effects of polymerization on membrane fluidity have not been investigated. Here planar supported lipid bilayers (PSLBs) composed of dienoyl PCs on glass substrates were examined to assess the degree to which UV-initiated polymerization affects lateral lipid mobility. Fluorescence recovery after photobleaching (FRAP) was used to measure the diffusion coefficients (D) and mobile fractions of rhodamine-DOPE in unpolymerized and polymerized PSLBs composed of bis-sorbyl phosphatidylcholine (bis-SorbPC), mono-sorbyl-phosphatidylcholine (mono-SorbPC), bis-dienoyl-phosphatidylcholine (bis-DenPC), and mono-dienoyl phosphatidylcholine (mono-DenPC). Polymerization was performed in both the Lα and Lβ phase for each lipid. In all cases, polymerization reduced membrane fluidity; however, measurable lateral diffusion was retained which is attributed to a low degree of polymerization. The D values for sorbyl lipids were less than those of the denoyl lipids; this may be a consequence of the distal location of polymerizable group in the sorbyl lipids which may facilitate interleaflet bonding. The D values measured after polymerization were 0.1-0.8 of those measured before polymerization, a range that corresponds to fluidity intermediate between that of a Lα phase and a Lβ phase. This D range is comparable to ratios of D values reported for liquid-disordered (Ld) and liquid-ordered (Lo) lipid phases and indicates that the effect of UV polymerization on lateral diffusion in a dienoyl PSLB is similar to the transition from a Ld phase to a Lo phase. The partial retention of fluidity in UV-polymerized PSLBs, their enhanced stability, and the activity of incorporated transmembrane proteins and peptides is discussed.


The CDC13-STN1-TEN1 complex stimulates Pol α activity by promoting RNA priming and primase-to-polymerase switch.
Emerging evidence suggests that Cdc13-Stn1-Ten1 (CST), an RPA-like ssDNA-binding complex, may regulate primase-Pol α (PP) activity at telomeres constitutively, and at other genomic locations under conditions of replication stress. Here we examine the mechanisms of PP stimulation by CST using purified complexes derived from Candida glabrata. While CST does not enhance isolated DNA polymerase activity, it substantially augments both primase activity and primase-to-polymerase switching. CST also simultaneously shortens the RNA and lengthens the DNA in the chimeric products. Stn1, the most conserved subunit of CST, is alone capable of PP stimulation. Both the N-terminal OB fold and the C-terminal winged-helix domains of Stn1 can bind to the Pol12 subunit of the PP complex and stimulate PP activity. Our findings provide mechanistic insights on a well-conserved pathway of PP regulation that is critical for genome stability.


Complement C3a treatment accelerates recovery after stroke via modulation of astrocyte reactivity and cortical connectivity.
Despite advances in acute care, ischemic stroke remains a major cause of long-term disability. Approaches targeting both neuronal and glial responses are needed to enhance recovery and improve long-term outcome. The complement C3a receptor (C3aR) is a regulator of inflammation with roles in neurodevelopment, neural plasticity, and neurodegeneration. Using mice lacking C3aR (C3aR-/-) and mice overexpressing C3a in the brain, we uncovered 2 opposing effects of C3aR signaling on functional recovery after ischemic stroke: inhibition in the acute phase and facilitation in the later phase. Peri-infarct astrocyte reactivity was increased and density of microglia reduced in C3aR-/- mice; C3a overexpression led to the opposite effects. Pharmacological treatment of wild-type mice with intranasal C3a starting 7 days after stroke accelerated recovery of motor function and attenuated astrocyte reactivity without enhancing microgliosis. C3a treatment stimulated global white matter reorganization, increased peri-infarct structural connectivity, and upregulated Igf1 and Thbs4 in the peri-infarct cortex. Thus, C3a treatment from day 7 after stroke exerts positive effects on astrocytes and neuronal connectivity while avoiding the deleterious consequences of C3aR signaling during the acute phase. Intranasal administration of C3aR agonists within a convenient time window holds translational promise to improve outcome after ischemic stroke.


Probing the mechanics of aqueous two-phase extraction using large diameter single-walled carbon nanotubes.
We have demonstrated that large diameter (1.8 ± 0.4 nm) carbon nanotubes (CNTs) can be separated by means of the aqueous two-phase extraction (ATPE). This rapid and convenient tool has enabled us to isolate fractions of particular CNT diameter distribution. We have shown how a range of parameters can be used to fine tune the characteristics of the isolated material. Interestingly, by varying the pH of the medium, we have suppressed the extraction of low diameter CNTs and only large diameter CNTs were obtained. A number of other factors such as selected surfactant concentration steps, temperature or amount of starting CNT material have been found to have a significant effect on the end result of the CNT differentiation. The findings have provided us with more insight regarding the underlying mechanics of ATPE for processing polydisperse CNT mixtures.


β-Caryophyllene Liposomes Attenuate Neurovascular Unit Damage After Subarachnoid Hemorrhage in Rats.
This study was conducted to prepare β-caryophyllene loaded liposomes (BCP-LP) and investigated their effects on neurovascular unit (NVU) damage after subarachnoid hemorrhage (SAH) in rats. A blood injection into the pre-chiasmatic cistern was used to achieve SAH. BCP-LP were prepared, characterized and administrated to rats with SAH. The prepared BCP-LP were spherical with a size distribution of approximately 189.3 nm and Zeta potential of - 13.9 mV. Neurological scoring, the balance beam test, cerebral blood flow monitoring, brain edema and biochemical analyses were applied to evaluate the effects of BCP-LP on rat NVU damage after SAH. The results demonstrated that BCP-LP treatment improved neurological function disorder, balance ability and cerebral blood perfusion in rats. Brain edema detection and blood-brain barrier permeability detection revealed that BCP-LP could reduce brain edema and promote repairment of blood-brain barrier after SAH. Using the western blot experiments, we demonstrated that BCP-LP attenuated the loss of tight junction proteins Occludin and Zonula occludens-1, inhibit the high expression of VEGFR-2 and GFAP, and promote the repair of laminin. These results demonstrate the protective effect BCP-LP exert in the NVU after SAH in rats, and supports the use of BCP-LP for future study and therapy of SAH.


Femoral neck fractures after hip resurfacing.
Femoral neck fracture is an important early complication after hip resurfacing. Our aims were firstly to determine the incidence of fracture in an independent series and secondly, in a case control study, to investigate potential risk factors. Fifteen femoral neck fractures occurred in a series of 842 procedures, representing an incidence of 1.8%. No relationship existed between age, sex, and fracture incidence. Mechanical factors such as notching, femoral neck lengthening, and varus alignment of the femoral component were found to have a similar incidence in both fracture and control groups. The proportion of patients that had at least 1 mechanical risk factor was not different between the 2 groups (fracture group, 50%; control group, 41%). Established avascular necrosis of the femoral head was evident in all retrieved femoral heads (n = 9) of patients who sustained postoperative fracture; in none of these patients was avascular necrosis the initial diagnosis. This study suggests that in our practice, mechanical factors, such as neck notching, neck lengthening, or varus angulations, are not the primary cause of femoral neck fractures.


Long-term safety and efficacy of canakinumab in cryopyrin-associated periodic syndrome: results from an open-label, phase III pivotal study in Japanese patients.
To assess the long-term safety and efficacy of canakinumab in Japanese patients with cryopyrin-associated periodic syndrome (CAPS). In this open-label phase 3 study, Japanese patients aged ≥2 years with CAPS received canakinumab 2-8 mg/kg subcutaneously every 8 weeks. The duration of the core treatment phase was 24 weeks followed by 22 months extension phase. The primary objective was the proportion of patients free of clinical and serologic relapse at week 24. The study enrolled 19 Japanese patients (median age, 14 years; range, 2-48 years) with CAPS [MWS, 7 (36.8%); NOMID, 12 (63.2%)] for a median of 109 weeks. Fifteen patients (79%) achieved a complete response by day 15, 18 (94.7%) by week 24 and all by week 48. At the end of the study, 18 (95%) were free from relapse and 11 (57.9%) were assessed as having no disease activity by the PGA. Thirteen (68%) patients (MWS, 4; NOMID, 9) had their canakinumab dose increased during the trial. All patients experienced at least one adverse event (AE), the most common being infections (100%) and 5 (26.3%) reported serious AEs. No deaths were reported and the only patient who discontinued the study early withdrew consent. Regular canakinumab treatment every 8 weeks at dose levels from 2-8 mg/kg, based on the clinical need, represents a successful strategy to induce rapid and complete response while maintain long-term disease control in Japanese patients with CAPS. The safety profile of canakinumab was consistent with that observed from previous studies.


Methylation of discrete regions of the O6-methylguanine DNA methyltransferase (MGMT) CpG island is associated with heterochromatinization of the MGMT transcription start site and silencing of the gene.
O6-Methylguanine DNA methyltransferase (MGMT) repairs the mutagenic and cytotoxic O6-alkylguanine lesions produced by environmental carcinogens and the chemotherapeutic nitrosoureas. As such, MGMT-mediated repair of O6-alkylguanine lesions constitutes a major form of resistance to nitrosourea chemotherapy and makes control of MGMT expression of clinical interest. The variability of expression in cell lines and tissues, along with the ease with which the MGMT phenotype reverts under various conditions, suggests that MGMT is under epigenetic control. One such epigenetic mechanism, 5-methylation of cytosines, has been linked to MGMT expression. We have used an isogenic human multiple myeloma tumor cell line model composed of an MGMT-positive parent cell line, RPMI 8226/S, and its MGMT-negative variant, termed 8226/V, to study the control of MGMT expression. The loss of MGMT activity in 8226/V was found to be due to the loss of detectable MGMT gene expression. Bisulfite sequencing of the MGMT CpG island promoter revealed large increases in the levels of CpG methylation within discrete regions of the 8226/V MGMT CpG island compared to those in 8226/S. These changes in CpG methylation are associated with local heterochromatinization of the 8226/V MGMT transcription start site and provide a likely mechanism for the loss of MGMT transcription in 8226/V.


Decline in the Tropospheric Abundance of Halogen from Halocarbons: Implications for Stratospheric Ozone Depletion
Analyses of air sampled from remote locations across the globe reveal that tropospheric chlorine attributable to anthropogenic halocarbons peaked near the beginning of 1994 and was decreasing at a rate of 25 ± 5 parts per trillion per year by mid-1995. Although bromine from halons was still increasing in mid-1995, the summed abundance of these halogens in the troposphere is decreasing. To assess the effect of this trend on stratospheric ozone, estimates of the future stratospheric abundance of ozone-depleting gases were made for mid-latitude and polar regions on the basis of these tropospheric measurements. These results suggest that the amount of reactive chlorine and bromine will reach a maximum in the stratosphere between 1997 and 1999 and will decline thereafter if limits outlined in the adjusted and amended Montreal Protocol on Substances That Deplete the Ozone Layer are not exceeded in future years.


Zebrafish heart regeneration occurs by cardiomyocyte dedifferentiation and proliferation.
Although mammalian hearts show almost no ability to regenerate, there is a growing initiative to determine whether existing cardiomyocytes or progenitor cells can be coaxed into eliciting a regenerative response. In contrast to mammals, several non-mammalian vertebrate species are able to regenerate their hearts, including the zebrafish, which can fully regenerate its heart after amputation of up to 20% of the ventricle. To address directly the source of newly formed cardiomyocytes during zebrafish heart regeneration, we first established a genetic strategy to trace the lineage of cardiomyocytes in the adult fish, on the basis of the Cre/lox system widely used in the mouse. Here we use this system to show that regenerated heart muscle cells are derived from the proliferation of differentiated cardiomyocytes. Furthermore, we show that proliferating cardiomyocytes undergo limited dedifferentiation characterized by the disassembly of their sarcomeric structure, detachment from one another and the expression of regulators of cell-cycle progression. Specifically, we show that the gene product of polo-like kinase 1 (plk1) is an essential component of cardiomyocyte proliferation during heart regeneration. Our data provide the first direct evidence for the source of proliferating cardiomyocytes during zebrafish heart regeneration and indicate that stem or progenitor cells are not significantly involved in this process.


Structure-Effect Relationships of Novel Semi-Synthetic Cannabinoid Derivatives.
Background: As a library of cannabinoid (CB) derivatives with (-)-trans-cannabidiol (CBD) or (-)-trans-cannabidivarin (CBDV) scaffold, we synthesized nine novel cannabinoids: 2-hydroxyethyl cannabidiolate (2-HEC), 2-hydroxypentyl cannabidiolate (2-HPC), 2,3-dihydroxypropyl cannabidiolate (GCBD), cyclohexyl cannabidiolate (CHC), n-hexyl-cannabidiolate (HC), 2-(methylsulfonamido)ethyl cannabidiolate (NMSC), 2-hydroxyethyl cannabidivarinolate (2-HECBDV), cyclohexyl cannabidivarinolate (CHCBDV), and n-hexyl cannabidivarinolate (HCBDV). Their binding and intrinsic effects at the CB1- and CB2-receptors and the effects on inflammatory signaling cascades were investigated in in vitro and ex vivo cell models. Materials and Methods: Binding affinity was studied in membranes isolated from CB-receptor-transfected HEK293EBNA cells, intrinsic functional activity in Chinese hamster ovary (CHO) cells, and activation of nuclear factor κB (NF-κB) and nuclear factor of activated T-cells (NFAT) in phorbol 12-myristate 13-acetate (PMA)/ionomycin (IO)-treated Jurkat T-cells. Inhibition of interleukin (IL)-17-induced pro-inflammatory cytokines and chemokines [IL-6, IL-1β, CC-chemokine ligand 2 (CCL2), and tumor necrosis factor (TNF)-α] was studied in RAW264.7 macrophages at the RNA level. Pro-inflammatory cytokine (IL-1β, IL-6, IL-8, and TNF-α) expression and prostaglandin E2 (PGE2) expression were investigated at the protein level in lipopolysaccharide (LPS)-treated primary human monocytes. Results: Derivatives with long aliphatic side chains at the ester position at R1 [HC (5)] as well as the ones with polar side chains [2-HECBDV (7), NMSC (6), and 2-HEC (1)] can be selective for CB2-receptors. The CBDV-derivatives HCBDV and CHCBDV demonstrated specific binding at CB1- and CB2-receptors at nanomolar concentrations. 2-HEC, 2-HPC, GCBD, and NMSC were agonists at CB2-receptor and antagonists at CB1-receptor. CHC bound both receptors at submicromolar ranges and was an agonist for these receptors. 2-HECBDV was an agonist at CB2-receptor and an antagonist at the CB1-receptor despite its modest affinity at this receptor (micromolar range). NMSC inhibited NF-κB and NFAT activity, and 2-HEC, 2-HPC, and GCBD dose-dependently inhibited PMA/IO-stimulated NFAT activation. CHC and HC dose-dependently reduced IL-1β and CCL2 messenger RNA (mRNA) expression. NMSC inhibited IL-1β, CCL2, and TNF-α at lower doses. At higher doses, it induced a pronounced increase in IL-6 mRNA. 2-HEC, 2-HPC, and GCBD dose-dependently inhibited LPS-induced IL-1β, TNF-α, and IL-6 synthesis. NMSC further increased LPS-stimulated IL-1β release but inhibited IL-8, TNF-α, and PGE2. Conclusion: The CBD- and CBDV-derivatives studied are suitable for targeting CB-receptors. Some may be used as selective CB2 agonists. The length of the aliphatic rest at R2 of CBD (pentyl) and CBDV (propyl) did not correlate with the binding affinity. Higher polarity at R1 appeared to favor the agonistic activity at CB2-receptors.


Randomised double-blind comparative study of dexmedetomidine and tramadol for post-spinal anaesthesia shivering.
Dexmedetomidine (α2 adrenergic agonist) has been used for prevention of post anaesthesia shivering. Its use for the treatment of post-spinal anaesthesia shivering has not been evaluated. The aim of this study was to evaluate and compare the efficacy, haemodynamic and adverse effects of dexmedetomidine with those of tramadol, when used for control of post-spinal anaesthesia shivering. A prospective, randomised, and double-blind study was conducted in 50 American Society of Anaesthesiologists Grade I and II patients of either gender, aged between 18 and 65 years, scheduled for various surgical procedures under spinal anaesthesia. The patients were randomised in two groups of 25 patients each to receive either dexmedetomidine 0.5 μg/kg or tramadol 0.5 mg/kg as a slow intravenous bolus. Grade of shivering, onset of shivering, time for cessation of shivering, recurrence, response rate, and adverse effects were observed at scheduled intervals. Unpaired t-test was used for analysing the data. Time taken for cessation of shivering was significantly less with dexmedetomidine when compared to tramadol. Nausea and vomiting was observed only in tramadol group (28% and; 20% respectively). There was not much difference in the sedation profile of both the drugs. We conclude that although both drugs are effective, the time taken for cessation of shivering is less with dexmedetomidine when compared to tramadol. Moreover, dexmedetomidine has negligible adverse effects, whereas tramadol is associated with significant nausea and vomiting.


Wernicke's encephalopathy in a patient with gastric carcinoma: a diagnosis not to miss.
We describe a case of a patient who presented with a 20-day history of vomiting, generalised weakness and loss of appetite and a 2-day history of altered sensorium. On examination, he was grossly emaciated and there were no palpable lymph nodes. Central nervous system examination revealed nystagmus with bilateral lateral recti palsy and abdominal examination showed mild hepatomegaly. MRI of the brain showed bilateral and symmetrical hypertense signal changes in T2-weighted and fluid-attenuated inversion recovery sequences with diffusion restriction in the paramedian ventromedial thalamus. These findings were compatible with Wernicke's encephalopathy. He was started on thiamine supplementation with which neurological signs improved. An ultrasound of the abdomen showed mild hepatomegaly with multiple hyperechoic lesions and wall thickening of the pyloric antrum. Upper gastroduodenoscopy showed ulcerative lesions involving the antrum, pylorus and duodenum. Biopsy revealed moderately differentiated adenocarcinoma. The patient underwent palliative gastrojejunostomy and was clinically better at discharge. It is important to consider Wernicke encephalopathy in patients with gastric cancer who have acute neurological symptoms.


Prevalence of modalities of headaches and bruxism among patients with craniomandibular disorder.
This study compared the presence of headache and bruxing behavior among 133 craniomandibular disorder patients (CMD) referred to the The Center For the Study Of Craniomandibular Disorders and to the presence of headache and bruxing behavior occurring in 133 controls seeking routine dental care. Both patients and controls were consecutive referrals to the clinic occurring over a three year period. The mean age of the CMD group was 38 years (range 28-42), and the mean age of the controls was 37 years (range 25-44). The information gathered included questionnaire and clinical examination. Different types of headaches, signs and symptoms of CMD, and bruxing behavior were assessed both in the CMD group and in the corresponding control group. Results of this study showed that bruxing behavior and headache pain were significantly more prevalent in the CMD group (57%, 76%) than in the corresponding control group (37%, 49%). Of the three types of headache observed, tension and combination headaches were more prevalent in the CMD group (n = 48 = 36% and n = 37 = 28%). Migraine headache was more prevalent in the CMD group (n = 16 = 12%) than in the control group (n = 3 = 2%). It was concluded that headache and bruxing behavior predominated in CMD patients. This data reinforces the need to assess headache pain and signs and symptoms of bruxing behavior in CMD patients, particularly in those suffering chronic facial pain and headache.


Clinical diagnosis and management of suspicious pigmented skin lesions--a survey of GPs.
The ability of general practitioners to make important clinical decisions about the diagnosis and management of skin lesions is poorly understood. A questionnaire on the diagnosis and management of eight photographed skin lesions was sent to 150 GPs in southeast Queensland. The questionnaire was completed by 114 GPs (response rate 77%). General practitioners' provisional diagnoses and management of photographed skin lesions were mostly or always correct, and there was general high consistency between diagnosis and intended management. Pigmented seborrhoeic keratoses were the most difficult lesions for GPs to diagnose correctly. Whether a lesion was different to usual moles appears to have the strongest association with clinical diagnosis. The high ability of GPs as measured in this artificial study is encouraging. The strong association between identifying moles that appear different to usual and correct clinical diagnoses suggest that unless GPs can increase the number of skin lesions they see as part of their typical workload, their clinical ability may not increase further.


Has brain imaging discovered anything new about how the brain works?
There have now been roughly 130,000 papers on fMRI. While these have clearly contributed to our understanding of the functional anatomy of the human brain, it is less clear that they have changed the way in which we think about the brain. The issue, in other words, is whether they have established new principles about how the brain works. In this paper we offer as an example one new principle, partly to lay down the criteria that are required for establishing a new principle, and partly to encourage others to offer other principles. Our example concerns the flexible flow of information through the cortex that must occur according to the demands of the task or current context. We suggest that this flexibility is achieved by feedback connections from the prefrontal and parietal cortex, and that these include connections to sensory and motor areas. However, the nature of the selective effect differs. The parietal cortex can select both within and across processing streams. By across streams we mean that it can have the same influence on different streams, for example the dorsal and ventral visual systems. However, only the prefrontal cortex can also select between processing streams. The difference between the prefrontal and parietal effects is due to their different positions within the processing hierarchy.


Human Pluripotent Stem Cell-Derived Neural Cells and Brain Organoids Reveal SARS-CoV-2 Neurotropism.
Neurological complications are common in patients with COVID-19. While SARS-CoV-2, the causal pathogen of COVID-19, has been detected in some patient brains, its ability to infect brain cells and impact their function are not well understood, and experimental models using human brain cells are urgently needed. Here we investigated the susceptibility of human induced pluripotent stem cell (hiPSC)-derived monolayer brain cells and region-specific brain organoids to SARS-CoV-2 infection. We found modest numbers of infected neurons and astrocytes, but greater infection of choroid plexus epithelial cells. We optimized a protocol to generate choroid plexus organoids from hiPSCs, which revealed productive SARS-CoV-2 infection that leads to increased cell death and transcriptional dysregulation indicative of an inflammatory response and cellular function deficits. Together, our results provide evidence for SARS-CoV-2 neurotropism and support use of hiPSC-derived brain organoids as a platform to investigate the cellular susceptibility, disease mechanisms, and treatment strategies for SARS-CoV-2 infection.


Clinical and endocrinological investigations on superovulation induced in heifers by human menopausal gonadotrophin.
After an i.v. injection of human menopausal gonadotrophin (hMG), three components of the disappearance time in heifers were found for immunoreactive LH (half-life 13.8--1020 min) and immunoreactive FSH (half-life 21.3--1090 min). When heifers were treated daily with hMG from Days 9, 10 or 11 of the cycle for 3 or 5 days, a total dose of 1350 i.u. FSH induced 10.2 +/- 2.5 (mean +/- s.d.) corpora lutea (CL) in 6 animals and 13 persistent follicles in 4 animals, while a total dose of 2100 i.u. FSH induced 14.3 +/- 1.5 CL in 6 heifers and there were no persistent follicles. Nine heifers treated with a single i.m. injection of 1500 i.u. PMSG exhibited 11.4 +/- 8.6 CL with 17 persistent follicles in 7 animals. Progesterone concentrations rose significantly faster and the oestradiol drop was more rapid after oestrus in heifers treated with hMG than in those treated with PMSG. These results demonstrate that stimulation of the ovarian follicles of heifers is more homogeneous when hMG is used.


A new family of plant antifungal proteins.
Plant seeds contain high concentrations of many antimicrobial proteins. These include chitinases, beta-1,3-glucanases, proteinase inhibitors, and ribosome-inactivating proteins. We recently reported the presence in corn seeds of zeamatin, a protein that has potent activity against a variety of fungi but has none of the above activities. Zeamatin is a 22-kDa protein that acts by causing membrane permeabilization Using a novel bioautography technique, we found similar antifungal proteins in the seeds of 6 of 12 plants examined. A polyclonal antiserum was raised against zeamatin and was used in immunoblots to confirm the presence of zeamatinlike proteins in these seeds. N-terminal amino acid sequencing was carried out on the antifungal proteins from corn, oats, sorghum, and wheat, and these sequences revealed considerable homology with each other. Interestingly, these N-terminal sequences are also similar to those of thaumatin, a pathogenesis-related protein from tobacco, and two salt stress-induced proteins. These results indicate that zeamatin is not unique but is a member of a previously unrecognized family of plant defense proteins that may include some species of pathogenesis-related proteins.


Long-term effects of a toddler-focused caries prevention programme among Northwestern US tribal children: The TOTS-to-Tweens study.
We sought to determine whether American Indian tribe-based interventions that successfully prevented toddler dental caries in a 2005 cohort study (the Toddler Overweight and Tooth Decay Prevention Study, or TOTS) influenced the prevalence of dental caries in children ages 11 to 13 in the same communities ten years later (the TOTS-to-Tweens study). We recruited original TOTS participants and conducted school- and community-based dental screenings at tribal communities that received family plus community-wide interventions (F + CW), community interventions only (CW) or were control communities. We also enrolled children who did not participate in TOTS, but were exposed to CW interventions or to the control environment. Trained clinicians examined children's teeth and recorded whether each tooth was decayed, missing or filled (DMFT). We calculated DMFT scores for each child and evaluated differences in DMFT incidence rate ratios (IRR) and components of DMFT by intervention group. We observed lower age- and sex-adjusted DMFT scores among F + CW children (a mean of 2.1 DMFT; 95% confidence interval [CI]: 1.4-2.7) and among CW children (2.2; 95% CI: 1.9-2.6), than control children (3.0; 95% CI: 2.3-3.7). The F + CW group had 32% lower DMFT scores than control children (IRR = 0.68; 95% CI: 0.46-1.01), and CW children had 26% lower DMFT scores than control (IRR = 0.74; 95% CI: 0.55-1.00). The proportion of children with filled teeth was higher in control than intervention communities (37.9% in F + CW, 47.1% in CW, and 67.1% in control, P = .002). Our findings suggest modest yet significant long-term effects of interventions that prevented toddler dental caries on the DMFT scores of tweens evaluated ten years later. Further study of effective interventions and their sustainability is clearly warranted among tribal children, who remain at high risk for dental caries.


Cross-reactions between streptococcal M proteins and human transplantation antigens.
Allogeneic antiserums against human lymphocytes were specifically inhibited by M protein from beta hemolytic group A Streptococcus pyogenes, type 1. Analogous M proteins from streptococcus types 3, 4, 5, 6, 12, and 14 had little or no inhibitory activity. The specific inhibition by M protein is not a result of anticomplementary activity or of coating of the lymphocyte surface. Streptococcal polysaccharide and 73 other polysaccharides were inactive. Because all seven HL-A specificities tested were inhibited, it is inferred that M1 protein has a structure common to human histocompatibility antigens.


Trading legitimacy: everyday corruption and its consequences for medical regulation in southern Vietnam.
Government regulation of health professionals is believed to ensure the efficacy and expertise of practitioners for and on behalf of patients. Certification and licensing are two common means to do so, legalizing a physician to practice medicine. However, ethnography from Ho Chi Minh City (HCMC) suggests that in corrupt socioeconomic environments, certification and licensing can alternatively produce a trade in legitimacy. Drawing on participant observations during 15 months of fieldwork with 25 medical acupuncturists in private practice in HCMC, southern Vietnam, and their patients, I argue that everyday practices of corruption and the importance of personal networks meant that legality, efficacy, and expertise separated. Certificates and licenses did not unproblematically validate expertise and efficacy. Consequently, compliance and enforcement of regulations as solutions to inadequate medical care may not achieve the effects intended.


Stereoselective synthesis of the head group of archaeal phospholipid PGP-Me to investigate bacteriorhodopsin-lipid interactions.
Phosphatidylglycerophosphate methyl ester (PGP-Me), a major constituent of the archaeal purple membrane, is essential for the proper proton-pump activity of bacteriorhodopsin (bR). We carried out the first synthesis of the bisphosphate head group of PGP-Me using H-phosphonate chemistry that led to the production of a simplified PGP-Me analogue with straight alkyl chains. To investigate the role of this head group in the structural and functional integrity of bR, the analogue was used to reconstitute bR into liposomes, in which bR retained the original trimeric structure and light-induced photocycle activity. Enhanced ordering of an alkyl chain of the (2)H-labelled analogue was observed in (2)H NMR spectra upon interaction with bR. These results together suggest that the bisphosphate moiety plays a role in the proper functioning of bR through the lipid-protein interaction.


Branhamella catarrhalis: epidemiology, surface antigenic structure, and immune response.
Over the past decade, Branhamella catarrhalis has emerged as an important human pathogen. The bacterium is a common cause of otitis media in children and of lower respiratory tract infections in adults with chronic obstructive pulmonary disease. B. catarrhalis is exclusively a human pathogen. It colonizes the respiratory tract of a small proportion of adults and a larger proportion of children. Studies involving restriction enzyme analysis of genomic DNA show that colonization is a dynamic process, with the human host eliminating and acquiring new strains frequently. The surface of B. catarrhalis contains outer membrane proteins, lipooligosaccharide, and pili. The genes which encode several outer membrane proteins have been cloned, and some of these proteins are being studied as potential vaccine antigens. Analysis of the immune response has been limited by the lack of an adequate animal model of B. catarrhalis infection. New information regarding outer membrane structure should guide studies of the human immune response to B. catarrhalis. Immunoassays which specifically detect antibodies to determinants exposed on the bacterial surface will elucidate the most relevant immune response. The recognition of B. catarrhalis as an important human pathogen has stimulated research on the epidemiology and surface structures of the bacterium. Future studies to understand the mechanisms of infection and to elucidate the human immune response to infection hold promise of developing new methods to treat and prevent infections caused by B. catarrhalis.


Common components of evidence-based parenting programs for preventing maltreatment of school-age children.
Child maltreatment can lead to a variety of negative outcomes in childhood including physical and mental health problems that can extend into adulthood. Given the transactional nature of child maltreatment and the difficulties that many maltreating families experience, child protection services typically offer various kinds of programs to maltreated children, their parents, and/or their families. Although the specific difficulties experienced by these families may vary, sub-optimal parenting practices are typically part of the picture and may play a central role in maltreated children's development. Hence, to deal with child maltreatment, programs that focus on parenting practices are essential, and identifying the common components of effective programs is of critical importance. The objectives of the present study were to: 1) describe the components of evidence-based parenting programs aimed at parents who have maltreated their elementary school-aged children or are at-risk for doing so and 2) identify the components that are common to these programs, using the approach proposed by Barth and Liggett-Creel (2014). Fourteen evidence-based parenting programs aimed at parents who had maltreated their elementary school-aged children (ages 6-12) or were at-risk for doing so were identified using both a review of relevant online databases of evidence-based programs (California Evidence-Based Clearinghouse for Child Welfare, Blueprints for Healthy Youth Development, Youth.gov, and the National Registry of Evidence-based Programs and Practices). Common components were identified (operationalized as components present in two thirds of programs) and discussed. The identification of common components of evidence-based programs may help clinicians choose the best intervention methods.


Comparison of telomere length in black and white teachers from South Africa: the sympathetic activity and ambulatory blood pressure in Africans study.
Telomere length is a marker of biological aging that has been linked to cardiovascular disease risk. The black South African population is witnessing a tremendous increase in the prevalence of cardiovascular disease, part of which might be explained through urbanization. We compared telomere length between black South Africans and white South Africans and examined which biological and psychosocial variables played a role in ethnic difference in telomere length. We measured leukocyte telomere length in 161 black South African teachers and 180 white South African teachers aged 23 to 66 years without a history of atherothrombotic vascular disease. Age, sex, years having lived in the area, human immunodeficiency virus (HIV) infection, hypertension, body mass index, dyslipidemia, hemoglobin A1c, C-reactive protein, smoking, physical activity, alcohol abuse, depressive symptoms, psychological distress, and work stress were considered as covariates. Black participants had shorter (median, interquartile range) relative telomere length (0.79, 0.70-0.95) than did white participants (1.06, 0.87-1.21; p < .001), and this difference changed very little after adjusting for covariates. In fully adjusted models, age (p < .001), male sex (p = .011), and HIV positive status (p = .023) were associated with shorter telomere length. Ethnicity did not significantly interact with any covariates in determining telomere length, including psychosocial characteristics. Black South Africans showed markedly shorter telomeres than did white South African counterparts. Age, male sex, and HIV status were associated with shorter telomere length. No interactions between ethnicity and biomedical or psychosocial factors were found. Ethnic difference in telomere length might primarily be explained by genetic factors.


A meta-analysis of cardiac electrophysiology computational models.
Computational models of cardiac electrophysiology are exemplar demonstrations of the integration of multiple data sets into a consistent biophysical framework. These models encapsulate physiological understanding to provide quantitative predictions of function. The combination or extension of existing models within a common framework allows integrative phenomena in larger systems to be investigated. This methodology is now routinely applied, as demonstrated by the increasing number of studies which use or extend previously developed models. In this study, we present a meta-analysis of this model re-use for two leading models of cardiac electrophysiology in the form of parameter inheritance trees, a sensitivity analysis and a comparison of the functional significance of the sodium potassium pump for defining restitution curves. These results indicate that even though the models aim to represent the same physiological system, both the sources of parameter values and the function of equivalent components are significantly different.


Processes in the development of international specialist competencies and standards: the Sports Physiotherapy for All Project.
In a world of rapidly developing knowledge it is important that professions describe their roles and capabilities. The need for a thorough description of sports physiotherapy was addressed through collaboration between the International Federation of Sports Physiotherapy (IFSP) and five European higher education institutions. This resulted in the Sports Physiotherapy for All Project, which has been successful in developing internationally accepted competencies and standards for sports physiotherapists. This article describes and reflects on the process to communicate useful lessons. A competency model was chosen to facilitate differentiation and communication of aspects of sports physiotherapy practice. Documentation relating to sports physiotherapy practice was collected from 16 countries and analysed thematically. A cut and paste method was used by a panel of experts to allocate themes to areas of practice within the competency model. Theme groups were used to select areas of practice for description in competency form. Standards were derived from competencies following in depth discussion with the expert panel, and triangulation with themes derived from international documentation. A rigorous process of international review and revision led to the final list of 11 competencies and related standards, both accepted by the IFSP. This work provides a foundation for the development of an audit toolkit to guide demonstration and evaluation of competencies and standards. This provides a foundation for targeted career development activities, appropriate provision of training opportunities, and quality enhancement. The experiences gained during this project can inform other health professions and their specialisms when embarking on a similar journey.


Isolation of DNA probes specific for rat chromosomal regions 19p, 19q and 4q and their application for the analysis of diethylstilbestrol-induced aneuploidy in binucleated rat fibroblasts.
DNA probes specific for rat chromosomes 19p, 19q and 4q were isolated, characterized and used for the detection and analysis of diethylstilbestrol(DES)-induced aneuploidy. By denaturing and partially reassociating total genomic DNA a new rat repetitive DNA family was isolated, which was located on chromosome 19p21. Sequencing of a number of subclones from cos76-1 and other clones of this so-called 76-family revealed that the repeat units are interrupted with large areas of other (unique) DNA. Consequently, after fluorescence in situ hybridization (FISH) the signals in interphase nuclei are large and spread out. The other two probes, cos25 (chromosome 4q) and cos42-47 (chromosome 19q), were isolated by screening cosmid libraries with probes isolated previously in our laboratory. The repeat unit of cos25 is a 2174 bp long EcoRI unit that contains three Sau3A sites and is tandemly organized. Sequencing of subclones of cos42-47 revealed that this probe was in fact the 5S RNA gene, located on 19q12. In order to determine if these probes were suitable probes for aneuploidy detection, two series of dual colour FISH with the combinations cos25/cos76-1 (4q/19p) and cos42-47/cos76-1 (19q/19p) were carried out on slides from an in vitro micronucleus assay with DES. With all three probes used, an increase in binucleated cells with non-disjunction or chromosome loss was observed in the DES-treated cultures. Scoring of additional micronucleated cells on slides hybridized with the cos25/cos76-1 (4q/19p) probes revealed that the hybridization signal of probe cos25 (4q) was over-represented in the micronuclei of the control cultures. The simultaneous use of the 19q and 19p probes is a particularly valuable tool for the detection of aneuploidy, since it allows distinction between aneugenic and clastogenic events in binucleated cells. Results of this analysis showed that apart from aneuploidy, DES also induced structural chromosome aberrations, although to a lesser extent.


18FDG PET in hallucinating and non-hallucinating patients.
The aim of the study was to detect whether the abnormal regional brain activity correlates with auditory verbal hallucination-proneness (AVH) in a group of patients with schizophrenia and schizophrenia-related psychoses. 15 patients with prominent AVH (score for hallucination intensity--item 3 in the PANSS > or =4) and 15 control patients without AVH (item 3 PANSS score < or =2) underwent 18FDG positron emission tomography at rest. SPM group analysis revealed an increased uptake of 18FDG in the right middle frontal gyrus (BA46) in subjects with high verbal hallucination score compared to non-hallucinating patients (p<0.001, uncorrected). Activation in BA46 positively correlated with the intensity of hallucinations (Spearman r=0.57; p<0.001). The observed functional recruitment of the right prefrontal cortex in subjects with high hallucination score may reflect impairment in the integration of intended actions and sensory feedback resulting in misattribution of internal events to an external source. This mechanism may form the cognitive basis for AVH.


Effect of photoactivation systems and resin composites on the microleakage of esthetic restorations.
The aim of this study was to evaluate the influence of four photoactivation systems [quartz tungsten halogen (QTH), light-emitting diode (LED), argon ion laser (AL), and plasma arc curing PAC)] on cementum/dentin and enamel microleakage of Class II restorations using a microhybrid [Z250-3M ESPE] and two packable composites [(SureFil-Dentsply and Tetric Ceram HB-Ivoclair/Vivadent]. Three hundred sixty "vertical-slot Class II cavities" were prepared at the mesial surface of bovine incisors using a 245 carbide bur in a highspeed handpiece. Specimens were divided into twelve groups (composite-photoactivation systems). Half of the specimens had the gingival margin placed in enamel (n=15) and the other half in cementum/dentin (n=15). Composites were inserted and cured in 2 mm increments according to manufacturers' recommended exposure times. After polishing, the samples were immersed in 2% methylene blue solution, sectioned, and evaluated at the gingival margins. Data were submitted to statistical analysis using the Kruskal-Wallis and Mann-Whitney tests. No significant differences were found among the photoactivation systems and among resin composites (p>0.05). Microleakage was not significantly affected by location (enamel vs. cementum/dentin, p>0.05). These findings suggested neither the photoactivation systems nor the resin composite types might have an effect on the microleakage at gingival margins Class II cavities.


Ray Tracing Structured AMR Data Using ExaBricks.
Structured Adaptive Mesh Refinement (Structured AMR) enables simulations to adapt the domain resolution to save computation and storage, and has become one of the dominant data representations used by scientific simulations; however, efficiently rendering such data remains a challenge. We present an efficient approach for volume- and iso-surface ray tracing of Structured AMR data on GPU-equipped workstations, using a combination of two different data structures. Together, these data structures allow a ray tracing based renderer to quickly determine which segments along the ray need to be integrated and at what frequency, while also providing quick access to all data values required for a smooth sample reconstruction kernel. Our method makes use of the RTX ray tracing hardware for surface rendering, ray marching, space skipping, and adaptive sampling; and allows for interactive changes to the transfer function and implicit iso-surfacing thresholds. We demonstrate that our method achieves high performance with little memory overhead, enabling interactive high quality rendering of complex AMR data sets on individual GPU workstations.


EVEC, a novel epidermal growth factor-like repeat-containing protein upregulated in embryonic and diseased adult vasculature.
A hallmark of vascular lesions is the phenotypic modulation of vascular smooth muscle cells (VSMCs) from a quiescent, contractile state to a more primitive, proliferative phenotype with a more fetal pattern of gene expression. Using subtraction hybridization to identify genes that may regulate this transition, we cloned a novel gene named EVEC, an acronym for its expression in the embryonic vasculature and the presence of Ca2+ binding epidermal growth factor-like repeats contained in the predicted protein structure. Although these repeats are characteristic of the extracellular matrix proteins, fibrillin, fibulin, and the latent transforming growth factor-beta binding proteins, EVEC most closely resembles the H411 and T16/S1-5 gene products, the latter of which are believed to regulate DNA synthesis in quiescent fibroblasts. Using in situ hybridization, we demonstrated that EVEC is expressed predominantly in the VSMCs of developing arteries in E11.5 through E16.5 mouse embryos. Lower levels of expression are also observed in endothelial cells, perichondrium, intestine, and mesenchyme of the face and kidney. EVEC mRNA expression is dramatically downregulated in adult arteries, except in the uterus, where cyclic angiogenesis continues; however, EVEC expression is reactivated in 2 independent rodent models of vascular injury. EVEC mRNA is observed in cellular elements of atherosclerotic plaques of LDL receptor-deficient, human apolipoprotein B transgenic mice and in VSMCs of the media and neointima of balloon-injured rat carotid arteries. These data suggest that EVEC may play an important role in the regulation of vascular growth and maturation during development and in lesions of injured vessels.


Prediction of postoperative pulmonary complications in a population-based surgical cohort.
Current knowledge of the risk for postoperative pulmonary complications (PPCs) rests on studies that narrowly selected patients and procedures. Hypothesizing that PPC occurrence could be predicted from a reduced set of perioperative variables, we aimed to develop a predictive index for a broad surgical population. Patients undergoing surgical procedures given general, neuraxial, or regional anesthesia in 59 hospitals were randomly selected for this prospective, multicenter study. The main outcome was the development of at least one of the following: respiratory infection, respiratory failure, bronchospasm, atelectasis, pleural effusion, pneumothorax, or aspiration pneumonitis. The cohort was randomly divided into a development subsample to construct a logistic regression model and a validation subsample. A PPC predictive index was constructed. Of 2,464 patients studied, 252 events were observed in 123 (5%). Thirty-day mortality was higher in patients with a PPC (19.5%; 95% [CI], 12.5-26.5%) than in those without a PPC (0.5%; 95% CI, 0.2-0.8%). Regression modeling identified seven independent risk factors: low preoperative arterial oxygen saturation, acute respiratory infection during the previous month, age, preoperative anemia, upper abdominal or intrathoracic surgery, surgical duration of at least 2 h, and emergency surgery. The area under the receiver operating characteristic curve was 90% (95% CI, 85-94%) for the development subsample and 88% (95% CI, 84-93%) for the validation subsample. The risk index based on seven objective, easily assessed factors has excellent discriminative ability. The index can be used to assess individual risk of PPC and focus further research on measures to improve patient care.


Tissue-type plasminogen activator is a regulator of monocyte diapedesis through the brain endothelial barrier.
Inflammatory cell trafficking into the brain complicates several neurological disorders including multiple sclerosis. Normally, reliable brain functioning is maintained and controlled by the blood-brain barrier (BBB), which is essential to restrict the entry of potentially harmful molecules and cells from the blood into the brain. The BBB is a selective barrier formed by dedicated brain endothelial cells and dependent on the presence of intracellular tight junctions. In multiple sclerosis, a severe dysfunction of the BBB is observed, which is key to monocyte infiltration and inflammation in the brain. Proteolytic activity has been associated with these inflammatory processes in the brain. Our studies in plasma of rats indicated that the extracellular protease tissue-type plasminogen activator (tPA) correlates with the clinical signs of experimental allergic encephalomyelitis, a rat model of multiple sclerosis. In this study, we studied the function of the tPA during diapedesis of monocytes through a rat and human brain endothelial barrier. Monocyte-brain endothelial cell coculture experiments showed that monocytes induce the release of tPA by brain endothelial cells, which subsequently activates the signal transduction protein extracellular signal related kinase (ERK1/2), both involved in monocyte diapedesis. Importantly, live imaging and immunoblot analyses of rat brain endothelial cells revealed that tPA and ERK1/2 control the breakdown of the tight junction protein occludin. These studies identify tPA as a novel and relevant pathological mediator of neuroinflammation and provide a potential mechanism for this.


Nonrandom sampling in human genetics: familial correlations.
In the context of human genetics, sampling is often nonrandom in that pedigrees are frequently selected by virtue of their having at least one affected individual. For quantitative traits, it may be that the proband is selected because of a phenotypic value above some predetermined cut-point; this will also be true for diseases defined by a cut-point above which individuals are said to be affected (e.g. diabetes, hypertension, and obesity). Relatively little attention has been given to the implications of these forms of nonrandom sampling. In this paper, we show that the biases introduced by such sampling are sufficient to lead to erroneous model specification and biased parameter estimates in path analysis. Existing methodologies used to correct for such sampling (e.g., elimination of proband or regression techniques) also result in similar bias in estimation and model fitting. Thus, it is possible to make inferences which do not reflect underlying biology, but are artifacts of the sampling design. As a consequence, method-of-moment estimators are developed for means, variances, and correlations under such sampling. Simulation results are used to demonstrate the superiority of this method over alternate strategies.


Post-ruminal non-protein nitrogen supplementation as a strategy to improve fibre digestion and N efficiency in the ruminant.
The ruminant is able to transform plant fibres and non-protein nitrogen (NPN) into edible foods for human consumption. In an effort towards improving our understanding of this process, we sought to challenge convention and examine how the source, amount and site of NPN delivery in the gastrointestinal tract of the ruminant may affect fibre digestibility, rumen stability and N metabolism. In the first study presented here, we used four ruminally cannulated non-lactating heifers in a Latin square design to infuse 59 g/days of N in the form of ammonia (A) or urea (U) into either the rumen (R) or the abomasum (A). We found that intake was higher (p = .03) for animals receiving supplementary NPN as urea compared with ammonia. In addition, abomasally infused urea (UA) improved fibre fermentation by 9.4% (p = .05) and decreased ruminal pH fluctuations (lower slope in the cumulative pH parameters, p = .04) when compared with the same amount of urea infused ruminally (UR). In a second study, using the same group of heifers, we infused 50 or 150 g/day of urea into the rumen (UR50 and UR150) or 150 g of urea in the abomasum (UA150) or 50 g in the rumen and 100 g in the abomasum (URA150). Fibre digestion was improved by 4% (p = .02) when the same dose of urea was infused into the abomasum compared with the rumen, while estimated microbial protein production and N efficiency were not different between a low rumen dose and high post-ruminal dose of urea. Collectively, these studies provide insight into the viability of post-ruminal NPN supplementation as a strategy to improve fibre digestion and NPN inclusion in the ruminant diet.


End-of-Life Care Planning: The Importance of Older Adults' Marital Status and Gender.
Background: As the U.S. population ages, dramatic shifts are occurring in the proportion of older adults who are divorced and widowed. Health status and behaviors are known to differ across marital status groups, yet research on end-of-life (EOL) care planning has only compared married and unmarried persons, overlooking differences between divorced and widowed individuals, by gender. Objective: This study aimed to examine marital status differences in EOL care planning by comparing the likelihood of discussions about EOL care, designation of medical durable power of attorney (MDPOA) for health care decisions, and completion of a living will for married, divorced, and widowed older adults, by gender. Methods: Analyses used data from the U.S. Health and Retirement Study for 2243 adults (50 years of age and older), who died during the course of the study. Post-death, proxy respondents reported on the decedents' EOL care discussions, living will completion, and establishment of an MDPOA. Multivariate regressions were estimated to test differences in care planning across marital status groups, for men and women. Results: Divorced men were less likely than married men to have had care discussions and to have engaged in any of the three planning behaviors. Widowers were more likely to have established an MDPOA. Both divorced and widowed women were more likely to have performed any of these EOL planning activities than married women. Conclusions: Divorced men and married women are at risk for lacking EOL care planning. Practitioners are encouraged to discuss the importance of such planning as they encounter these at-risk groups.


Endurance Exercise Increases Intestinal Uptake of the Peanut Allergen Ara h 6 after Peanut Consumption in Humans.
Controlled studies on the effect of exercise on intestinal uptake of protein are scarce and underlying mechanisms largely unclear. We studied the uptake of the major allergen Ara h 6 following peanut consumption in an exercise model and compared this with changes in markers of intestinal permeability and integrity. Ten overnight-fasted healthy non-allergic men (n = 4) and women (n = 6) (23 ± 4 years) ingested 100 g of peanuts together with a lactulose/rhamnose (L/R) solution, followed by rest or by 60 min cycling at 70% of their maximal workload. Significantly higher, though variable, levels of Ara h 6 in serum were found during exercise compared to rest (Peak p = 0.03; area under the curve p = 0.006), with individual fold changes ranging from no increase to an increase of over 150-fold in the uptake of Ara h 6. Similarly, uptake of lactulose (2-18 fold change, p = 0.0009) and L/R ratios (0.4-7.9 fold change, p = 0.04) were significantly increased which indicates an increase in intestinal permeability. Intestinal permeability and uptake of Ara h 6 were strongly correlated (r = 0.77, p < 0.0001 for lactulose and Ara h 6). Endurance exercise after consumption may lead to increased paracellular intestinal uptake of food proteins.


Comprehensive analysis of SMAD4 mutations and protein expression in juvenile polyposis: evidence for a distinct genetic pathway and polyp morphology in SMAD4 mutation carriers.
Juvenile polyposis syndrome (JPS; OMIM 174900) is a rare disorder which is characterized by the presence of hamartomatous polyps throughout the gastrointestinal tract and an increased risk of gastrointestinal malignancy. Mutations of the SMAD4 gene on chromosome 18q21.1 have been shown to cause a subset of JPS cases, with estimates ranging from 20% to >50%. Characterization of the genes that cause the remainder of JPS cases relies on the certainty that SMAD4 is not the causative gene. We have undertaken a comprehensive analysis of germline SMAD4 mutations in a cohort of JPS patients to define the spectrum of mutations that cause JPS. We have analyzed a series of polyps from these patients for SMAD4 protein expression. We have also performed a blinded assessment of polyp material to look for morphological differences between polyps from patients with and without a germline SMAD4 mutation. The results indicate that almost all germline SMAD4 mutations are readily detectable by screening genomic DNA using polymerase chain reaction-based methods; SMAD4 can be excluded as the causative gene in the majority of our JPS cohort. Loss of SMAD4 expression occurs in most polyps from SMAD4 mutation carriers, even those with missense germline mutations. SMAD4 loss in polyps is, however, not a feature of cases that are not caused by SMAD4 mutations, indicating that these polyps develop along a SMAD4-independent pathway. The morphology of polyps from SMAD4 mutation carriers is subtly different from other JPS polyps, notably including a more prominent epithelial component in the former.


The environmental impact of sewage and wastewater outfalls in Antarctica: An example from Davis station, East Antarctica.
We present a comprehensive scientific assessment of the environmental impacts of an Antarctic wastewater ocean outfall, at Davis station in East Antarctica. We assessed the effectiveness of current wastewater treatment and disposal requirements under the Protocol on Environmental Protection to the Antarctic Treaty. Macerated wastewater has been discharged from an outfall at Davis since the failure of the secondary treatment plant in 2005. Water, sediment and wildlife were tested for presence of human enteric bacteria and antibiotic resistance mechanisms. Epibiotic and sediment macrofaunal communities were tested for differences between sites near the outfall and controls. Local fish were examined for evidence of histopathological abnormalities. Sediments, fish and gastropods were tested for uptake of sewage as measured by stable isotopes of N and C. Escherichia coli carrying antibiotic resistance determinants were found in water, sediments and wildlife (the filter feeding bivalve Laternula eliptica). Fish (Trematomus bernacchii) within close proximity to the outfall had significantly more severe and greater occurrences of histopathological abnormalities than at controls, consistent with exposure to sewage. There was significant enrichment of 15N in T. bernacchii and the predatory gastropod Neobuccinum eatoni around the outfall, providing evidence of uptake of sewage. There were significant differences between epibiotic and sediment macrofaunal communities at control and outfall sites (<1.5 km), when sites were separated into groups of similar habitat types. Benthic community composition was also strongly related to habitat and environmental drivers such as sea ice. The combined evidence indicated that the discharge of wastewater from the Davis outfall is causing environmental impacts. These findings suggest that conditions in Antarctic coastal locations, such as Davis, are unlikely to be conducive to initial dilution and rapid dispersal of wastewater as required under the Protocol on Environmental Protection to the Antarctic Treaty. Current minimum requirements for wastewater treatment and disposal in Antarctica are insufficient to ameliorate these risks and are likely to lead to accumulation of contaminants and introduction of non-native microbes and associated genetic elements. This new understanding suggests that modernised approaches to the treatment and disposal of wastewater are required in Antarctica. The most effective solution is advanced levels of wastewater treatment, which are now possible, feasible and a high priority for installation. As a direct outcome of the study, a new advanced treatment system is being installed at Davis, effectively avoiding environmental risks.


Construction of a soluble adenylyl cyclase activated by Gs alpha and forskolin.
A soluble adenylyl cyclase was constructed by linkage of portions of the cytosolic domains of the mammalian type I and type II enzymes. The soluble enzyme was stimulated by both forskolin and the alpha subunit of the heterotrimeric guanine nucleotide-binding protein (G protein) Gs (Gs alpha). Expression of the construct complemented the catabolic defect in a strain of Escherichia coli that is deficient in adenylyl cyclase activity. The active, approximately 60-kilodalton enzyme accumulated in the cytoplasmic fraction of E. coli to yield activities in excess of 1 nanomole per minute per milligram of protein. The two sets of transmembrane helices of mammalian adenylyl cyclases are thus not necessary for the catalytic or the most characteristic regulatory activities of the enzyme. This system may be useful for both genetic and biochemical analysis of G protein-regulated adenylyl cyclases.


Causal links between dorsal medial superior temporal area neurons and multisensory heading perception.
The dorsal medial superior temporal area (MSTd) in the extrastriate visual cortex is thought to play an important role in heading perception because neurons in this area are tuned to both optic flow and vestibular signals. MSTd neurons also show significant correlations with perceptual judgments during a fine heading direction discrimination task. To test for a causal link with heading perception, we used microstimulation and reversible inactivation techniques to artificially perturb MSTd activity while monitoring behavioral performance. Electrical microstimulation significantly biased monkeys' heading percepts based on optic flow, but did not significantly impact vestibular heading judgments. The latter result may be due to the fact that vestibular heading preferences in MSTd are more weakly clustered than visual preferences and multiunit tuning for vestibular stimuli is weak. Reversible chemical inactivation, however, increased behavioral thresholds when heading judgments were based on either optic flow or vestibular cues, although the magnitude of the effects was substantially stronger for optic flow. Behavioral deficits in a combined visual/vestibular stimulus condition were intermediate between the single-cue effects. Despite deficits in discrimination thresholds, animals were able to combine visual and vestibular cues near optimally, even after large bilateral muscimol injections into MSTd. Simulations show that the overall pattern of results following inactivation is consistent with a mixture of contributions from MSTd and other areas with vestibular-dominant tuning for heading. Our results support a causal link between MSTd neurons and multisensory heading perception but suggest that other multisensory brain areas also contribute.


Antibacterial activity of BMS-180680, a new catechol-containing monobactam.
The in vitro activities of a new catechol-containing monobactam, BMS-180680 (SQ 84,100), were compared to those of aztreonam, ceftazidime, imipenem, piperacillin-tazobactam, ciprofloxacin, amikacin, and trimethoprim-sulfamethoxazole. BMS-180680 was often the most active compound against many species of the family Enterobacteriaceae, with MICs at which 90% of the isolates were inhibited (MIC90s) of < or = 0.5 microg/ml for Escherichia coli, Klebsiella spp., Citrobacter diversus, Enterobacter aerogenes, Serratia marcescens, Proteus spp., and Providencia spp. BMS-180680 had moderate activities (MIC90s of 2 to 8 microg/ml) against Citrobacter freundii, Morganella morganii, Shigella spp., and non-E. aerogenes Enterobacter spp. BMS-180680 was the only antibiotic evaluated that was active against >90% of the Pseudomonas aeruginosa (MIC90, 0.25 microg/ml), Burkholderia cepacia, and Stenotrophomonas maltophilia (MIC90s, 1 microg/ml) strains tested. BMS-180680 was inactive against most strains of Pseudomonas fluorescens, Pseudomonas stutzeri, Pseudomonas diminuta, and Burkholderia pickettii. BMS-180680 was moderately active (MIC90s of 4 to 8 microg/ml) against Alcaligenes spp. and Acinetobacter lwoffii and less active (MIC90, 16 microg/ml) against Acinetobacter calcoaceticus-Acinetobacter baumanii complex. BMS-180680 lacked activity against gram-positive bacteria and anaerobic bacteria. Both tonB and cir fiu double mutants of E. coli had greatly decreased susceptibility to BMS-180680. Of the TEM, PSE, and chromosomal-encoded beta-lactamases tested, only the K1 enzyme hydrolyzed BMS-180680 to any measurable extent. Like aztreonam, BMS-180680 bound preferentially to penicillin-binding protein 3. The MICs of BMS-180680 were not influenced by the presence of hematin or 5% sheep blood in the test medium or with incubation in an atmosphere containing 5% CO2. BMS-180680 MICs obtained under strict anaerobic conditions were significantly higher than those obtained in ambient air.


Cacophonies of aid, failed state building and NGOs in Haiti: setting the stage for disaster, envisioning the future.
The January 2010 earthquake in Haiti was a catastrophe not only for the loss of life it caused, but also because it destroyed the very thin layer of state administrative capacity that was in place in the country. This article argues that the fragility of the Haitian state institutions was exacerbated by international strategies that promoted NGOs as substitutes for the state. These strategies have generated a vicious circle that, while solving immediate logistical problems, ended up weakening Haiti's institutions. However, the article does not call for an overarching condemnation of NGOs. Instead, it explores two cases of community-based NGOs, Partners In Health and Fonkoze, that have contributed to creating durable social capital, generated employment and provided functioning services to the communities where they operated. The article shows that organisations that are financially independent and internationally connected, embrace a needs-based approach to their activities and share a long-term commitment to the communities within which they operate can contribute to bringing about substantial improvement for people living in situations of extreme poverty. It concludes that in the aftermath of a crisis of the dimension of the January earthquake it is crucial to channel support towards organisations that show this type of commitment.


Inkjet-printed microelectrodes on PDMS as biosensors for functionalized microfluidic systems.
Microfluidic systems based on polydimethylsiloxane (PDMS) have gained popularity in recent years. However, microelectrode patterning on PDMS to form biosensors in microchannels remains a worldwide technical issue due to the hydrophobicity of PDMS and its weak adhesion to metals. In this study, an additive technique using inkjet-printed silver nanoparticles to form microelectrodes on PDMS is presented. (3-Mercaptopropyl)trimethoxysilane (MPTMS) was used to modify the surface of PDMS to improve its surface wettability and its adhesion to silver. The modified surface of PDMS is rendered relatively hydrophilic, which is beneficial for the silver droplets to disperse and thus effectively avoids the coalescence of adjacent droplets. Additionally, a multilevel matrix deposition (MMD) method is used to further avoid the coalescence and yield a homogeneous pattern on the MPTMS-modified PDMS. A surface wettability comparison and an adhesion test were conducted. The resulting silver pattern exhibited good uniformity, conductivity and excellent adhesion to PDMS. A three-electrode electrochemical biosensor was fabricated successfully using this method and sealed in a PDMS microchannel, forming a lab-on-a-chip glucose biosensing system.


ApoER2-Dab1 disruption as the origin of pTau-related neurodegeneration in sporadic Alzheimer's disease.
Sporadic Alzheimer's disease (sAD) is not a global brain disease. Specific regions, layers and neurons degenerate early while others remain untouched even in advanced disease. The prevailing model used to explain this selective neurodegeneration-prion-like Tau spread-has key limitations and is not easily integrated with other defining sAD features. Instead, we propose that in humans Tau hyperphosphorylation occurs locally via disruption in ApoER2-Dab1 signaling and thus the presence of ApoER2 in neuronal membranes confers vulnerability to degeneration. Further, we propose that disruption of the Reelin/ApoE/ApoJ-ApoER2-Dab1-P85α-LIMK1-Tau-PSD95 (RAAAD-P-LTP) pathway induces deficits in memory and cognition by impeding neuronal lipoprotein internalization and destabilizing actin, microtubules, and synapses. This new model is based in part on our recent finding that ApoER2-Dab1 disruption is evident in entorhinal-hippocampal terminal zones in sAD. Here, we hypothesized that neurons that degenerate in the earliest stages of sAD (1) strongly express ApoER2 and (2) show evidence of ApoER2-Dab1 disruption through co-accumulation of multiple RAAAD-P-LTP components. We applied in situ hybridization and immunohistochemistry to characterize ApoER2 expression and accumulation of RAAAD-P-LTP components in five regions that are prone to early pTau pathology in 64 rapidly autopsied cases spanning the clinicopathological spectrum of sAD. We found that: (1) selectively vulnerable neuron populations strongly express ApoER2; (2) numerous RAAAD-P-LTP pathway components accumulate in neuritic plaques and abnormal neurons; and (3) RAAAD-P-LTP components were higher in MCI and sAD cases and correlated with histological progression and cognitive deficits. Multiplex-IHC revealed that Dab1, pP85αTyr607, pLIMK1Thr508, pTau and pPSD95Thr19 accumulated together within dystrophic dendrites and soma of ApoER2-expressing neurons in the vicinity of ApoE/ApoJ-enriched extracellular plaques. These observations provide evidence for molecular derangements that can be traced back to ApoER2-Dab1 disruption, in each of the sampled regions, layers, and neuron populations that are prone to early pTau pathology. Findings support the RAAAD-P-LTP hypothesis, a unifying model that implicates dendritic ApoER2-Dab1 disruption as the major driver of both pTau accumulation and neurodegeneration in sAD. This model provides a new conceptual framework to explain why specific neurons degenerate and identifies RAAAD-P-LTP pathway components as potential mechanism-based biomarkers and therapeutic targets for sAD.