First Report of Frogeye Leaf Spot of Soybean Caused by Cercospora sojina Race 11 in Virginia.
Frogeye leaf spot of soybean (FLS) (Glycine max (L.) Merr.), caused by Cercospora sojina Hara, was first detected in Virginia in 1942 (1). During the 2008 growing season, a FLS survey was conducted in soybean fields in Virginia. This was the first FLS race survey conducted in Virginia. Typical frogeye leaf spot symptoms, as reported by Phillips (4), were observed on soybean leaves in Westmoreland County. During 2008, Westmoreland County planted 7,365 ha of soybean. Symptomatic leaves were collected from V06-1891, V06-1365, V05-4394, V04-8405, and Hutcheson cultivars from plants in growth stages R5 to R6. Leaves were placed in a moist chamber for 24 h at 21°C with 12-h light to induce sporulation. C. sojina was only recovered from V06-1365. Conidia were removed from the leaves, placed into V8 juice agar amended with rifampicin (10 mg ml-1) and ampicillin (0.25 g liter-1) and incubated at 21°C with 12-h light. Cultures with dark pigmentation and presence of conidia were observed after 3 weeks. Conidia matched the description of C. sojina (4). Conidia had three to nine septa, were hyaline, elongate to fusiform, and measured 3 to 6 × 25 to 40 μm. Race identification was conducted using the set of differentials reported by Mian et al. (3). Spores for inoculation were produced on soybean stem lima bean agar (SSLBA) media. Tencentimeter-diameter pots, each containing four plants, were used. The test was conducted twice in a complete randomized design with three replications. Seedlings were inoculated at the V3 growth stage with a spore suspension of 6 × 104 spores/ml. Control plants were sprayed with sterile distilled water. Plants were placed in a greenhouse bench humidity chamber at 21°C for 72 h. Disease rating was conducted 14 days after inoculation. Since the resistance to FLS is known to be controlled by single dominant genes, the FLS was scored as a qualitative trait (i.e., resistant versus susceptible) as previously done by Mian et al. (2). Plants that showed numerous, large, spreading lesions were classified as susceptible and each plant was given a score of 1. Plants that showed no lesions or only small lesions or flecks were classified as resistant and each plant was given a score of 0. Control plants remained healthy. On the basis of the reaction response of the isolate on the set of differentials and comparison with the proposed race designations of Mian et al. (3), the isolate was classified as race 11. Race 11 shows compatible reaction (susceptibility) on the soybean cv. Lincoln, which is the source of Rcs1 resistance gene, and incompatible reactions (resistance) on cvs. Peking, Davis, and Kent. The latter two cultivars are sources of the Rcs3 and Rcs2 genes, respectively. Successful development of soybean cultivars with FLS resistance not only depends on knowledge of the presence of resistance genes, but also on the understanding of the pathogen population structure. To our knowledge this is the first report of C. sojina race 11 from soybean in Virginia. Resistance to this race is conditioned by Rcs2, Rcs3, and the single dominant gene in Peking (3). We recommend use of Rcs3 and Rcs2 genes and the single dominant gene in Peking for resistance to FLS in Virginia. References: (1) S. B. Fenn. Plant Dis. Rep. 26:383, 1942. (2) M. A. R. Mian et al. Crop Sci. 39:1687, 1999. (3) M. A. R. Mian et al. Crop Sci. 48:14, 2008. (4) D. V. Phillips. Page 20 in: Compendium of Soybean Diseases. 4th ed. The American Phytopathological Society. St. Paul, MN, 1999.


SKCa channels mediate the medium but not the slow calcium-activated afterhyperpolarization in cortical neurons.
Many neurons, including pyramidal cells of the cortex, express a slow afterhyperpolarization (sAHP) that regulates their firing. Although initial findings suggested that the current underlying the sAHP could be carried through SK(Ca) channels, recent work has uncovered anomalies that are not congruent with this idea. Here, we used overexpression and dominant-negative strategies to assess the involvement of SK(Ca) channels in mediating the current underlying the sAHP in pyramidal cells of the cerebral cortex. Pyramidal cells of layer V exhibit robust AHP currents composed of two kinetically and pharmacologically distinguishable currents known as the medium AHP current (I(mAHP)) and the slow AHP current (I(sAHP)). I(mAHP) is blocked by the SK(Ca) channel blockers apamin and bicuculline, whereas I(sAHP) is resistant to these agents but is inhibited by activation of muscarinic receptors. To test for a role for SK(Ca) channels, we overexpressed K(Ca)2.1 (SK1) and K(Ca)2.2 (SK2), the predominant SK(Ca) subunits expressed in the cortex, in pyramidal cells of cultured brain slices. Overexpression of K(Ca)2.1 and K(Ca)2.2 resulted in a fourfold to fivefold increase in the amplitude of I(mAHP) but had no detectable effect on I(sAHP). As an additional test, we examined I(sAHP) in a transgenic mouse expressing a truncated SK(Ca) subunit (SK3-1B) capable of acting as a dominant negative for the entire family of SK(Ca)-IK(Ca) channels. Expression of SK3-1B profoundly inhibited I(mAHP) but again had no discernable effect on I(sAHP). These results are inconsistent with the proposal that SK(Ca) channels mediate I(sAHP) in pyramidal cells and indicate that a different potassium channel mediates this current.


Utility and accuracy of template-directed dye-terminator incorporation with fluorescence-polarization detection for genotyping single nucleotide polymorphisms.
There are little independent data available about how well single nucleotide polymorphism (SNP) genotyping technologies perform in the typical molecular genetics laboratory. We evaluated the utility and accuracy of a widely used technology, template-directed dye-terminator incorporation with fluorescence-polarization detection (FP-TDI), in a sample of 177 SNPs selected solely on the basis of map location. Genotypes were generated without optimization using standard protocols. Overall, 81% of the SNPs we studied generated readable genotypes by FP-TDI. Thirty-two SNPs were genotyped in duplicate by PCR-RFLP orfluorescent dye-terminator sequencing. Out of a total of 631 duplicate genotypes, no true discrepancies were detected. The true error rate has a 95% chance of lying between 0 and 6 out of 1000 genotypes. We also tested for deviations from Hardy-Weinberg Equilibrium in 33 SNPs genotyped in 50 unrelated individuals, and no significant deviations were detected. Our FP-TDI data were readily adaptable to automated genotype calling using our own method of cluster analysis, which assigns a probability score to each genotype call. We conclude that FP-TDI is both efficient and accurate. The method can easily fill the needs of SNP genotyping projects at the scale typically used for regional or candidate-gene association studies.


Angiotensin receptor blocker protection against podocyte-induced sclerosis is podocyte angiotensin II type 1 receptor-independent.
In the present study, we tested the hypothesis that the renoprotective effect of an angiotensin receptor blocker depends on the angiotensin II type 1 (AT(1)) receptor on podocytes. For this purpose, we generated podocyte-specific knockout mice for the AT(1) gene (Agtr1a) and crossed with NEP25, in which selective podocyte injury can be induced by immunotoxin, anti-Tac(Fv)-PE38. Four weeks after the addition of anti-Tac(Fv)-PE38, urinary albumin:creatinine ratio was not attenuated in Agtr1a knockout/NEP25 mice (n=18) compared with that in control NEP25 mice (n=13; 8.08+/-2.41 in knockout versus 4.84+/-0.73 in control). Both strains of mice showed similar degrees of sclerosis (0.66+/-0.17 versus 0.82+/-0.27 on a 0 to 4 scale) and downregulation of nephrin (5.78+/-0.45 versus 5.65+/-0.58 on a 0 to 8 scale). In contrast, AT(1) antagonist or an angiotensin I-converting enzyme inhibitor, but not hydralazine, remarkably attenuated proteinuria and sclerosis in NEP25 mice. Moreover, continuous angiotensin II infusion induced microalbuminuria similarly in both Agtr1a knockout and wild-type mice. Thus, angiotensin inhibition can protect podocytes and prevent the development of glomerulosclerosis independent of podocyte AT(1). Possible mechanisms include inhibitory effects on AT(1) of other cells or through mechanisms independent of AT(1). Our study further demonstrates that measures that directly affect only nonpodocyte cells can have beneficial effects even when sclerosis is triggered by podocyte-specific injury.


An atlas and functional analysis of G-protein coupled receptors in human islets of Langerhans.
G-protein coupled receptors (GPCRs) regulate hormone secretion from islets of Langerhans, and recently developed therapies for type-2 diabetes target islet GLP-1 receptors. However, the total number of GPCRs expressed by human islets, as well as their function and interactions with drugs, is poorly understood. In this review we have constructed an atlas of all GPCRs expressed by human islets: the 'islet GPCRome'. We have used this atlas to describe how islet GPCRs interact with their endogenous ligands, regulate islet hormone secretion, and interact with drugs known to target GPCRs, with a focus on drug/receptor interactions that may affect insulin secretion. The islet GPCRome consists of 293 GPCRs, a majority of which have unknown effects on insulin, glucagon and somatostatin secretion. The islet GPCRs are activated by 271 different endogenous ligands, at least 131 of which are present in islet cells. A large signalling redundancy was also found, with 119 ligands activating more than one islet receptor. Islet GPCRs are also the targets of a large number of clinically used drugs, and based on their coupling characteristics and effects on receptor signalling we identified 107 drugs predicted to stimulate and 184 drugs predicted to inhibit insulin secretion. The islet GPCRome highlights knowledge gaps in the current understanding of islet GPCR function, and identifies GPCR/ligand/drug interactions that might affect insulin secretion, which are important for understanding the metabolic side effects of drugs. This approach may aid in the design of new safer therapeutic agents with fewer detrimental effects on islet hormone secretion.


PGR5-dependent cyclic electron transport around PSI contributes to the redox homeostasis in chloroplasts rather than CO(2) fixation and biomass production in rice.
The PGR5 (PROTON GRADIENT REGULATION 5) gene that is required for PSI cyclic electron transport in Arabidopsis was knocked down in rice (Oryza sativa). In three PGR5 knockdown (KD) lines, the PGR5 protein level was reduced to 5-8% of that in the wild type, resulting in a 50% reduction in PGRL1 (PGR5-LIKE PHOTOSYNTHETIC PHENOTYPE 1) protein levels. In ruptured chloroplasts, ferredoxin-dependent plastoquinone reduction activity was partially impaired; the phenotype was mimicked by addition of antimycin A to wild-type chloroplasts. As occurred in the Arabidopsis pgr5 mutant, non-photochemical quenching of Chl fluorescence (NPQ) induction was impaired in the leaves, but the electron transport rate (ETR) was only mildly affected at high light intensity. The P700(+) level was reduced even at low light intensity, suggesting that the PGR5 function was severely disturbed as in the Arabidopsis pgr5 mutant and that the other alternative routes of electrons could not compensate the stromal redox balance. The amplitude of the light-dark electrochromic shift (ECS) signal (ECSt), which reflects the total size of the proton motive force in steady-state photosynthesis, was reduced by 13-25% at approximately the growth light intensity. The CO(2) fixation rate was only slightly reduced in the PGR5 KD lines. Despite the drastic reduction in NPQ and P700(+) levels, total biomass was only slightly reduced in PGR5 KD lines grown at 370 µmol photons m(-2) s(-1). These results suggest that CO(2) fixation and growth rate are very robust in the face of alterations in the fundamental reactions of photosynthesis under constant light conditions in rice.


Efficacy of UV, acidified sodium hypochlorite, and mild heat for decontamination of surface and infiltrated Escherichia coli O157:H7 on green onions and baby spinach.
Produce-associated foodborne illnesses outbreaks have highlighted the need for more effective decontamination methods to ensure the safety of fresh produce. The main objective of this study was to evaluate the individual and combined efficacies of germicidal UV light (12.5 to 500 mJ/cm(2)), acidified sodium hypochlorite (ASC 10 to 200 ppm), and mild heat (40 to 50°C) for decontaminating green onions and baby spinach infected with Escherichia coli O157:H7. Samples were inoculated by spot and dip inoculation methods to mimic surface and infiltrated E. coli O157:H7 contamination, respectively. In green onions and baby spinach, the individual efficacies of UV, ASC, and mild-heat treatments varied based on the produce type and contamination method. Following analysis of the efficacies of the single treatments, a combined treatment with 125 mJ/cm(2) UV and 200 ppm of ASC at 50°C was selected for spot-inoculated green onions, and a combined treatment with 125 mJ/cm(2) UV and 200 ppm of ASC at 20°C was selected for spot- and dip-inoculated baby spinach. While a >5-log reduction was achieved with the combination treatment for spot-inoculated green onions with an initial contamination level of 7.2 log CFU per spot, the same treatment reduced E. coli O157:H7 populations below the detection limit (<1 log) on green onions spot inoculated at a lower contamination level (4.3 log CFU per spot). On spot- and dip-inoculated baby spinach, the combined treatment reduced E. coli O157:H7 populations by 2.8 log CFU per spot and 2.6 log CFU/g, respectively. The combined treatment of 500 mJ/cm(2) UV and 200 ppm of ASC at 50°C selected for the decontamination of dip-inoculated green onions resulted in a 2.2-log CFU/g reduction. These findings suggest that when foodborne pathogens contaminate produce and subsequently infiltrate, attach to, or become localized into protected areas, the individual or combined applications of UV, ASC, and mild-heat treatments have limited decontamination efficacies on both green onions and baby spinach (<3 log). However, treatments combining UV, ASC, and mild heat could be a promising application for reducing pathogen populations (>5 log) on E. coli O157:H7 surface-contaminated green onions. This study also highlights the importance of developing and optimizing produce-specific decontamination methods to ensure the safety of fresh produce commodities.


Nurturing care: promoting early childhood development.
The UN Sustainable Development Goals provide a historic opportunity to implement interventions, at scale, to promote early childhood development. Although the evidence base for the importance of early childhood development has grown, the research is distributed across sectors, populations, and settings, with diversity noted in both scope and focus. We provide a comprehensive updated analysis of early childhood development interventions across the five sectors of health, nutrition, education, child protection, and social protection. Our review concludes that to make interventions successful, smart, and sustainable, they need to be implemented as multi-sectoral intervention packages anchored in nurturing care. The recommendations emphasise that intervention packages should be applied at developmentally appropriate times during the life course, target multiple risks, and build on existing delivery platforms for feasibility of scale-up. While interventions will continue to improve with the growth of developmental science, the evidence now strongly suggests that parents, caregivers, and families need to be supported in providing nurturing care and protection in order for young children to achieve their developmental potential.


Research on heterodyne detection of a mode-locked pulse laser based on an acousto-optic frequency shift.
Heterodyne detection research is carried out on a mode-locked pulse laser using an acousto-optic frequency shifter (AOFS). Theoretical calculation and numerical simulation of the beat frequency of the mode-locked pulse were carried out, and the results show that the frequency offset can be calculated according to the beat period of the pulse waveform, and the offset is the frequency difference between the two adjacent longitudinal modes. Experiments were implemented with a self-built mode-locked laser using an AOFS to simulate the frequency shift of the detection laser induced by the object, the coherent beat frequency of the signal light and local oscillation light that were observed on the surface of the detector. The beat frequency wave was processed by filtering to obtain the frequency shift of the signal light. The experimental results are in good agreement with the numerical simulation.


"Akinetic depression" in schizophrenia.
It is possible that some "postpsychotic depressions" may be a toxic effect of antipsychotic drugs. Out of a total of 94 schizophrenic patients, 28 developed a mild akinesia and 32 never developed extrapyramidal symptoms. Those who developed akinesia became less psychotic, but they also experienced a significant, although modest, increase in depression ratings. Successful treatment of the akinesia resulted in significant improvements in depression, somatic concern, anxiety, emotional withdrawal, blunted affect, and motor retardation on both physicians' and nurses' ratings. A high association between akinesia and both objectively rated and subjectively experienced sedative effect indicates that an 'akinetic depression' is not likely if the patient does not look or feel drowsy. The 32 nonakinetic patients also became less psychotic, but not more depressed.


Retinoic acid synthesis and hindbrain patterning in the mouse embryo.
Targeted disruption of the murine retinaldehyde dehydrogenase 2 (Raldh2) gene precludes embryonic retinoic acid (RA) synthesis, leading to midgestational lethality (Niederreither, K., Subbarayan, V., Dolle, P. and Chambon, P. (1999). Nature Genet. 21, 444-448). We describe here the effects of this RA deficiency on the development of the hindbrain and associated neural crest. Morphological segmentation is impaired throughout the hindbrain of Raldh2-/- embryos, but its caudal portion becomes preferentially reduced in size during development. Specification of the midbrain region and of the rostralmost rhombomeres is apparently normal in the absence of RA synthesis. In contrast, marked alterations are seen throughout the caudal hindbrain of mutant embryos. Instead of being expressed in two alternate rhombomeres (r3 and r5), Krox20 is expressed in a single broad domain, correlating with an abnormal expansion of the r2-r3 marker Meis2. Instead of forming a defined r4, Hoxb1- and Wnt8A-expressing cells are scattered throughout the caudal hindbrain, whereas r5/r8 markers such as kreisler or group 3/4 Hox genes are undetectable or markedly downregulated. Lack of alternate Eph receptor gene expression could explain the failure to establish rhombomere boundaries. Increased apoptosis and altered migratory pathways of the posterior rhombencephalic neural crest cells are associated with impaired branchial arch morphogenesis in mutant embryos. We conclude that RA produced by the embryo is required to generate posterior cell fates in the developing mouse hindbrain, its absence leading to an abnormal r3 (and, to a lesser extent, r4) identity of the caudal hindbrain cells.


Body Patches in Inferior Temporal Cortex Encode Categories with Different Temporal Dynamics.
An unresolved question in cognitive neuroscience is how representations of object categories at different levels (basic and superordinate) develop during the course of the neural response within an area. To address this, we decoded categories of different levels from the spiking responses of populations of neurons recorded in two fMRI-defined body patches in the macaque STS. Recordings of the two patches were made in the same animals with the same stimuli. Support vector machine classifiers were trained at brief response epochs and tested at the same or different epochs, thus assessing whether category representations change during the course of the response. In agreement with hierarchical processing within the body patch network, the posterior body patch mid STS body (MSB) showed an earlier onset of categorization compared with the anterior body patch anterior STS body (ASB), irrespective of the categorization level. Decoding of the superordinate body versus nonbody categories was less dynamic in MSB than in ASB, with ASB showing a biphasic temporal pattern. Decoding of the ordinate-level category human versus monkey bodies showed similar temporal patterns in both patches. The decoding onset of superordinate categorizations involving bodies was as early as for basic-level categorization, suggesting that previously reported differences between the onset of basic and superordinate categorizations may depend on the area. The qualitative difference between areas in their dynamics of category representation may hinder the interpretation of decoding dynamics based on EEG or MEG, methods that may mix signals of different areas.


Management of a congenital tracheoesophageal fistula in a young Spanish water dog.
Tracheoesophageal fistula (TEF) in dogs is a rare disease with only few reports in the literature. This report aims to contribute to the current literature on suitable diagnostic methods for TEF and to provide follow-up information after successful surgical treatment. A seven-month-old intact female Spanish Water Dog was presented for further investigation of recurrent respiratory symptom. Bronchoscopy revealed a small hole-like defect in the tracheal wall at the bifurcation. The finding of the contrast material swallow study under fluoroscopy was indicative of a TEF. To further evaluate the connection between the trachea and esophagus, a computed tomography scan was performed. The TEF was surgically approached by thoracotomy through the right lateral sixth intercostal space. The fistula was identified, double ligated and divided. Histopathology confirmed the process to originate from the esophagus and to be patent. The dog was re-examined two weeks and ten months after surgery, with no evidence of recurring clinical signs. Contrast material swallow study using fluoroscopy was the most reliable diagnostic method. Bronchoscopy may allow the fistula to be visualized, but due to a small fistular opening it can lead to a false negative result. Surgical correction by ligation and dividing of the fistula suggests a good prognosis for early diagnosed and operated TEF.


The homeless in the emergency department: a patient profile.
To explore the possibility that homeless people use the accident and emergency (A&E) department as a substitute for primary care and to quantify the role of alcohol abuse in their attendances. The study period lasted six months. Attendances of all patients registering with "No fixed abode" as their address were analysed from the A&E notes. Information gathered included details of the presenting complaint, general practitioner (GP) registration, and alcohol involvement in the presentation. 135 homeless patients attended the department 233 times; 91% of patients were male with an average age of 40 years; 46% of attendances were during office hours; 81.5% of presentations were for minor problems; 65.2% of patients had co-existing medical problems, with 14.8% having a chronic alcohol problem; 23.7% of patients gave details of their GP and the remainder were either not registered or did not know their GP registration status; 29.6% of attendances directly involved alcohol and another 10.3% were requests for inpatient or outpatient alcohol detoxification; 42.1% of patients questioned said they were aware of the medical facilities available to homeless people in the community; 52.6% of those questioned said they preferred being seen in the A&E department when ill, with 23.7% preferring GP treatment and 10.5% attending community homeless clinics. The local homeless population may be using the A&E department as a substitute for primary care even in the presence of homeless healthcare facilities in the community. Heightened awareness of these facilities may improve their uptake. Alcohol plays a large role in homeless people seeking medical help in the A&E department. More accessible community facilities for dealing with this problem in this patient group are needed.


Structural characterization of EasH (Aspergillus japonicus) - an oxidase involved in cycloclavine biosynthesis.
Aj_EasH is a non-heme iron- and α-keto-glutarate-dependent oxidase that is responsible for an unusual cyclopropyl ring formation in the biosynthesis of the fungal ergot alkaloid cycloclavine. The three dimensional structure of Aj_EasH (2.2 Å resolution) reported here provides insight into the mechanism of this unusual and complex reaction.


Enhancement and inhibition of photolumincescence in hydrogenated amorphous silicon nitride microcavities.
A Fabry-Perot microcavity is used for the enhancement and inhibition of photoluminescence in hydrogenated amorphous silicon nitride. The amplitude of the photoluminescence is enhanced 4 times, while its linewidth is reduced 8 times with respect to the bulk hydrogenated amorphous silicon nitride. The transmittance, reflectance, and absorptance spectra of the microcavity were also measured and calculated. The calculated spectra agree well with the experimental ones.


Flufenamic acid protects against intestinal fluid secretion and barrier leakage in a mouse model of Vibrio cholerae infection through NF-κB inhibition and AMPK activation.
Nuclear factor kappa B (NF-κB)-mediated inflammatory responses play crucial roles in the pathogenesis of diarrhea caused by the Vibrio cholerae El Tor variant (EL), which is a major bacterial strain causing recent cholera outbreaks. Flufenamic acid (FFA) has previously been demonstrated to be a potent activator of AMP-activated protein kinase (AMPK), which is a negative regulator of NF-κB signaling. This study aimed to investigate the anti-diarrheal efficacy of FFA in a mouse model of EL infection and to investigate the mechanisms by which FFA activates AMPK in intestinal epithelial cells (IEC). In a mouse closed loop model of EL infection, FFA treatment (20mg/kg) significantly abrogated EL-induced intestinal fluid secretion and barrier disruption. In addition, FFA suppressed NF-κB nuclear translocation and expression of proinflammatory mediators and promoted AMPK phosphorylation in the EL-infected mouse intestine. In T84 cells, FFA induced AMPK activation. Furthermore, FFA promoted tight junction assembly and prevented interferon gamma (IFN-γ)-induced barrier disruption in an AMPK-dependent manner. Biochemical and molecular docking analyses indicated that FFA activates AMPK via a direct stimulation of calcium/calmodulin-dependent protein kinase kinase beta (CaMKKβ) activity. Collectively, our data indicate that FFA represents a class of existing drugs that may be of potential utility in the treatment of cholera caused by EL infection via AMPK-mediated suppression of NF-κB signaling in IEC.


Tunable nonlocal spin control in a coupled-quantum dot system.
The effective interaction between magnetic impurities in metals that can lead to various magnetic ground states often competes with a tendency for electrons near impurities to screen the local moment (known as the Kondo effect). The simplest system exhibiting the richness of this competition, the two-impurity Kondo system, was realized experimentally in the form of two quantum dots coupled through an open conducting region. We demonstrate nonlocal spin control by suppressing and splitting Kondo resonances in one quantum dot by changing the electron number and coupling of the other dot. The results suggest an approach to nonlocal spin control that may be relevant to quantum information processing.


Fabrication of three-dimensional scan-to-print ear model for microtia reconstruction.
Microtia is a congenital deformity of the external ear that occurs in 1 of every 5000 births. Microtia reconstruction using traditional two-dimensional templates does not provide highly detailed ear shapes. Here, we describe the feasibility of using a three-dimensional (3D) ear model as a reference. Seven children aged from 11 to 16 (6 grade III and 1 grade II microtia) were recruited from Seoul National University Children's Hospital, Korea. We generated 3D-computer-aided design models of each patient's ear by performing 3D laser scanning for a mirror-transformed cast of their normal ear. The 3D-printed ear model was used in microtia reconstruction surgery following the Nagata technique, and its shape was compared with the casted ear model. One patient experienced irritation caused by accidently pouring resin into the external auditory meatus, and another had minor skin necrosis; both complications were successfully treated. The average percentage differences of the superior, inferior, anterior, posterior, and lateral views between the casted and 3D-printed ear models were 1.17%, 1.48%, 1.64%, 1.80%, and 5.44%, respectively (average: 2.31%), where the difference between the casted ear models and traditional two-dimensional templates were 16.03% in average. Our results show that simple microtia reconstruction can be performed using 3D ear models. The 3D-printed ear models of each patient were consistent and accurately represented the thickness, depth, and height of the normal ear. The availability of the 3D-printed ear model in the operating room reduced the amount of unnecessary work during surgery.


Design and control of RUPERT: a device for robotic upper extremity repetitive therapy.
The structural design, control system, and integrated biofeedback for a wearable exoskeletal robot for upper extremity stroke rehabilitation are presented. Assisted with clinical evaluation, designers, engineers, and scientists have built a device for robotic assisted upper extremity repetitive therapy (RUPERT). Intense, repetitive physical rehabilitation has been shown to be beneficial overcoming upper extremity deficits, but the therapy is labor intensive and expensive and difficult to evaluate quantitatively and objectively. The RUPERT is developed to provide a low cost, safe and easy-to-use, robotic-device to assist the patient and therapist to achieve more systematic therapy at home or in the clinic. The RUPERT has four actuated degrees-of-freedom driven by compliant and safe pneumatic muscles (PMs) on the shoulder, elbow, and wrist. They are programmed to actuate the device to extend the arm and move the arm in 3-D space. It is very important to note that gravity is not compensated and the daily tasks are practiced in a natural setting. Because the device is wearable and lightweight to increase portability, it can be worn standing or sitting providing therapy tasks that better mimic activities of daily living. The sensors feed back position and force information for quantitative evaluation of task performance. The device can also provide real-time, objective assessment of functional improvement. We have tested the device on stroke survivors performing two critical activities of daily living (ADL): reaching out and self feeding. The future improvement of the device involves increased degrees-of-freedom and interactive control to adapt to a user's physical conditions.


Caregiver support groups: factors affecting use of services.
Use of time-limited psychoeducational support groups by family caregivers to help frail elderly people in the community reflects a mixture of predisposing, enabling, and need factors. Using multivariate regression methods that control for dependent-variable censoring, the authors found that attendance in support groups by primary caregivers was greater for those who were older, who had a secondary informal caregiver involved in providing care, or who had significant health problems. Attendance was greater also for those caring for Alzheimer's victims and for those experiencing higher levels of burden in their caregiving role. In English-language groups, Mexican American caregivers showed markedly lower attendance than Anglo American caregivers, but in Spanish-language groups attendance by Mexican American caregivers was essentially the same as that of the Anglo Americans in English-language groups. Data for this study are from a federally funded support group demonstration project conducted in a southwestern metropolitan area.


Optimization of pairings and detection conditions for measurement of FRET between cyan and yellow fluorescent proteins.
Detection of Förster resonance energy transfer (FRET) between cyan and yellow fluorescent proteins is a key method for quantifying dynamic processes inside living cells. To compare the different cyan and yellow fluorescent proteins, FRET efficiencies were measured for a set of the possible donor:acceptor pairs. FRET between monomeric Cerulean and Venus is more efficient than the ECFP:EYFP pair and has a 10% greater Förster distance. We also compared several live cell microscopy methods for measuring FRET. The greatest contrast for changes in intramolecular FRET is obtained using a combination of ratiometric and spectral imaging. However, this method is not appropriate for establishing the presence of FRET without extra controls. Accurate FRET efficiencies are obtained by fluorescence lifetime imaging microscopy, but these measurements are difficult to collect and analyze. Acceptor photobleaching is a common and simple method for measuring FRET efficiencies. However, when applied to cyan to yellow fluorescent protein FRET, this method becomes prone to an artifact that leads to overestimation of FRET efficiency and false positive signals. FRET was also detected by measuring the acceptor fluorescence anisotropy. Although difficult to quantify, this method is exceptional for screening purposes, because it provides high contrast for discriminating FRET.


The Tree Theme Method in psychosocial occupational therapy: a case study.
This study aimed to describe the Tree Theme Method (TTM) as a method for intervention in psychosocial occupational therapy. The TTM is based on theories concerning creative activities and occupational storytelling and story making. In order to exemplify the method a case study of a treatment process with follow up was undertaken. The participant was a female client suffering from anxiety and depression. During an interview the client painted symbolic trees on five different occasions with specific variations of the tree theme: a tree symbolizing her present life, her childhood, adolescence, adulthood, and, finally, a tree representing her future. The trees were used as starting points for the client to tell her life story. The intention was that she would find new strategies for how to change her daily life. Three years later there was a follow up stage where the client painted new trees and told her story. Some life themes were identified. The TTM appears suitable for intervention in psychosocial occupational therapy. In future studies the TTM should be subjected to evaluation research based on several clients in order to develop a deeper understanding of the process and what kind of results changes in the TTM intervention may provide.


Glutamine-, glutamine synthetase-, glutamate dehydrogenase- and pyruvate carboxylase-immunoreactivities in the rat dorsal root ganglion and peripheral nerve.
Supporting glial cells of the peripheral nervous system include satellite cells of dorsal root ganglia and Schwann cells of peripheral nerves. In the central nervous system, glial cells contain enzymes related to the tricarboxylic acid and glutamine cycles: pyruvate carboxylase, glutamate dehydrogenase, and glutamine synthetase. The present study used immunohistochemistry in the rat peripheral nervous system to determine the cellular distribution of these enzymes along with glutamine. In dorsal root ganglia and peripheral nerves, glutamine and glutamine related enzymes were enriched in satellite and Schwann cells. In the dorsal root ganglia, immunoreactive satellite cells surrounded neurons of all sizes. In peripheral nerve, immunoreactive Schwann cells were most easily observed surrounding large diameter, myelinated axons. These Schwann cells contained immunoreactivity in their cell bodies, nodes of Ranvier, and the rim of cytoplasm outside the myelin sheath. Myelin sheaths were non-immunoreactive. The peripheral glial tricarboxylic and glutamine cycles may be used to produce glutamine for neuronal cell uptake and conversion to glutamate for synaptic transmission. Alternatively, these cycles may function in peripheral glia similar to central nervous system astrocytes for supporting the energy demands of neurons.


Intracellular chloride channel protein CLIC1 regulates macrophage function through modulation of phagosomal acidification.
Intracellular chloride channel protein 1 (CLIC1) is a 241 amino acid protein of the glutathione S transferase fold family with redox- and pH-dependent membrane association and chloride ion channel activity. Whilst CLIC proteins are evolutionarily conserved in Metazoa, indicating an important role, little is known about their biology. CLIC1 was first cloned on the basis of increased expression in activated macrophages. We therefore examined its subcellular localisation in murine peritoneal macrophages by immunofluorescence confocal microscopy. In resting cells, CLIC1 is observed in punctate cytoplasmic structures that do not colocalise with markers for endosomes or secretory vesicles. However, when these macrophages phagocytose serum-opsonised zymosan, CLIC1 translocates onto the phagosomal membrane. Macrophages from CLIC1(-/-) mice display a defect in phagosome acidification as determined by imaging live cells phagocytosing zymosan tagged with the pH-sensitive fluorophore Oregon Green. This altered phagosomal acidification was not accompanied by a detectable impairment in phagosomal-lysosomal fusion. However, consistent with a defect in acidification, CLIC1(-/-) macrophages also displayed impaired phagosomal proteolytic capacity and reduced reactive oxygen species production. Further, CLIC1(-/-) mice were protected from development of serum transfer induced K/BxN arthritis. These data all point to an important role for CLIC1 in regulating macrophage function through its ion channel activity and suggest it is a suitable target for the development of anti-inflammatory drugs.


Dynamic Transmission Economic Evaluation of Infectious Disease Interventions in Low- and Middle-Income Countries: A Systematic Literature Review.
Economic evaluation using dynamic transmission models is important for capturing the indirect effects of infectious disease interventions. We examine the use of these methods in low- and middle-income countries, where infectious diseases constitute a major burden. This review is comprised of two parts: (1) a summary of dynamic transmission economic evaluations across all disease areas published between 2011 and mid-2014 and (2) an in-depth review of mosquito-borne disease studies focusing on health economic methods and reporting. Studies were identified through a systematic search of the MEDLINE database and supplemented by reference list screening. Fifty-seven studies were eligible for inclusion in the all-disease review. The most common subject disease was HIV/AIDS, followed by malaria. A diverse range of modelling methods, outcome metrics and sensitivity analyses were used, indicating little standardisation. Seventeen studies were included in the mosquito-borne disease review. With notable exceptions, most studies did not employ economic evaluation methods beyond calculating a cost-effectiveness ratio or net benefit. Many did not adhere to health care economic evaluations reporting guidelines, particularly with respect to full model reporting and uncertainty analysis. We present a summary of the state-of-the-art and offer recommendations for improved implementation and reporting of health economic methods in this crossover discipline.


Subjective tinnitus and its association with use of ear phones among students of the College of Medicine, University of Lagos, Nigeria.
The ear can be damaged by excessive noise levels. Noise can arise from variety of occupational/recreational sources causing tinnitus, hearing loss and hyperacusis. The use of different types of ear phones for entertainment has become very common/fashionable among our youths. This research aims to determine the prevalence of use of ear phones for entertainment and the prevalence of subjective tinnitus in students in an urban university setting; and the relationship between the two. This is a descriptive, cross sectional study of students of the college of medicine, University of Lagos, Nigeria from January to April 2012. A total of 388 willing participants (203 males, 185 females) were recruited by stratified random sampling from the 14 student hostel blocks. A self administered questionnaire was used. Data collected was analyzed using SPSS version 17.0.1. The prevalence of earphone use among the students and subjective tinnitus was 95.6% and 20.6% respectively. More than 90% of the ear phone users had duration of ear phone use of duration of 3 to 6 years.


Synthetic small molecules as anti-biofilm agents in the struggle against antibiotic resistance.
Biofilm formation significantly contributes to microbial survival in hostile environments and it is currently considered a key virulence factor for pathogens responsible for serious chronic infections. In the last decade many efforts have been made to identify new agents able to modulate bacterial biofilm life cycle, and many compounds have shown interesting activities in inhibiting biofilm formation or in dispersing pre-formed biofilms. However, only a few of these compounds were tested using in vivo models for their clinical significance. Contrary to conventional antibiotics, most of the anti-biofilm compounds act as anti-virulence agents as they do not affect bacterial growth. In this review we selected the most relevant literature of the last decade, focusing on the development of synthetic small molecules able to prevent bacterial biofilm formation or to eradicate pre-existing biofilms of clinically relevant Gram-positive and Gram-negative pathogens. In addition, we provide a comprehensive list of the possible targets to counteract biofilm formation and development, as well as a detailed discussion the advantages and disadvantages of the different current biofilm-targeting strategies.


The Anne Boleyn Illusion is a Six-Fingered Salute to Sensory Remapping.
The Anne Boleyn Illusion exploits the somatotopic representation of touch to create the illusion of an extra digit and demonstrates the instantaneous remapping of relative touch location into body-based coordinates through visuo-tactile integration. Performed successfully on thousands, it is also a simple demonstration of the flexibility of body representations for use at public events, in schools or in the home and can be implemented anywhere by anyone with a mirror and some degree of bimanual coordination.


Delayed parasite clearance after treatment with dihydroartemisinin-piperaquine in Plasmodium falciparum malaria patients in central Vietnam.
Reduced susceptibility of Plasmodium falciparum toward artemisinin derivatives has been reported from the Thai-Cambodian and Thai-Myanmar borders. Following increasing reports from central Vietnam of delayed parasite clearance after treatment with dihydroartemisinin-piperaquine (DHA-PPQ), the current first-line treatment, we carried out a study on the efficacy of this treatment. Between September 2012 and February 2013, we conducted a 42-day in vivo and in vitro efficacy study in Quang Nam Province. Treatment was directly observed, and blood samples were collected twice daily until parasite clearance. In addition, genotyping, quantitative PCR (qPCR), and in vitro sensitivity testing of isolates was performed. The primary endpoints were parasite clearance rate and time. The secondary endpoints included PCR-corrected and uncorrected cure rates, qPCR clearance profiles, in vitro sensitivity results (for chloroquine, dihydroartemisinin, and piperaquine), and genotyping for mutations in the Kelch 13 propeller domain. Out of 672 screened patients, 95 were recruited and 89 available for primary endpoint analyses. The median parasite clearance time (PCT) was 61.7 h (interquartile range [IQR], 47.6 to 83.2 h), and the median parasite clearance rate had a slope half-life of 6.2 h (IQR, 4.4 to 7.5 h). The PCR-corrected efficacy rates were estimated at 100% at day 28 and 97.7% (95% confidence interval, 91.2% to 99.4%) at day 42. At day 3, the P. falciparum prevalence by qPCR was 2.5 times higher than that by microscopy. The 50% inhibitory concentrations (IC50s) of isolates with delayed clearance times (≥ 72 h) were significantly higher than those with normal clearance times for all three drugs. Delayed parasite clearance (PCT, ≥ 72 h) was significantly higher among day 0 samples carrying the 543 mutant allele (47.8%) than those carrying the wild-type allele (1.8%; P = 0.048). In central Vietnam, the efficacy of DHA-PPQ is still satisfactory, but the parasite clearance time and rate are indicative of emerging artemisinin resistance. (This study has been registered at ClinicalTrials.gov under registration no. NCT01775592.).


Promoter hypermethylation in ductal carcinoma in situ of the male breast.
Ductal carcinoma in situ (DCIS) of the male breast is very rare and has hardly been studied molecularly. In males, we compared methylation status of 25 breast cancer-related genes in pure DCIS (n = 18) and invasive breast carcinoma (IBC) with adjacent DCIS (DCIS-AIC) (n = 44) using methylation-specific multiplex ligation-dependent probe amplification. Results were compared to female breast cancer (BC). There were no significant differences in methylation features between male pure DCIS, DCIS-AIC and IBC after correction for multiple comparisons. In paired analysis of IBC and adjacent DCIS, CADM1 showed a significantly higher absolute methylation percentage in DCIS (P = 0.002). In cluster analysis, two clusters stood out with respectively infrequent and frequent methylation (GATA5, KLLN, PAX6, PAX5, CDH13, MSH6 and WT1 were frequently methylated). Compared to female DCIS, methylation was in general much less common in male DCIS, especially for VHL, ESR1, CDKN2A, CD44, CHFR, BRCA2, RB1 and STK11. In contrast, THBS1 and GATA5 were more frequently methylated in male DCIS. In conclusion, there is frequent methylation of GATA5, KLLN, PAX6, PAX5, CDH13, MSH6 and WT1 in male DCIS. Since there was little change in the methylation status for the studied genes from pure male DCIS to DCIS-AIC and IBC, methylation of these seven genes is more likely to occur early in male breast carcinogenesis. Based on the current markers male DCIS seems to be an epigenetically more advanced precursor of male BC, although in comparison to its female counterpart it appears that fewer loci harbor methylation, pointing to differences between male and female breast carcinogenesis with regard to the studied loci.


Loss mechanisms in optical light pipes.
A geometrical optics approach is used to develop a theoretical model for analyzing loss mechanisms in optical light pipes. Five mechanisms are identified: intrinsic absorption, bulk scattering, losses that are due to roughness at the core-cladding interface, losses that are due to large-scale defects at the core-cladding interface, and losses that are due to absorption in the cladding material; and the effects of each of these on light-pipe transmission are considered. An approximate model appropriate for slightly rough surfaces is used to estimate the loss that is due to interface roughness. Optical experiments on commercially available light pipes are done to quantify the various loss processes. These experiments indicate that the interface effects play an important role in limiting the transmission in high-quality light pipes. From the optical measurements a rms interface roughness height in the 30-70-A range is deduced, and these values are confirmed by direct surface profilometry with an atomic force microscope.


Light at night acutely impairs glucose tolerance in a time-, intensity- and wavelength-dependent manner in rats.
Exposure to light at night (LAN) has increased dramatically in recent decades. Animal studies have shown that chronic dim LAN induced obesity and glucose intolerance. Furthermore, several studies in humans have demonstrated that chronic exposure to artificial LAN may have adverse health effects with an increased risk of metabolic disorders, including type 2 diabetes. It is well-known that acute exposure to LAN affects biological clock function, hormone secretion and the activity of the autonomic nervous system, but data on the effects of LAN on glucose homeostasis are lacking. This study aimed to investigate the acute effects of LAN on glucose metabolism. Male Wistar rats were subjected to i.v. glucose or insulin tolerance tests while exposed to 2 h of LAN in the early or late dark phase. In subsequent experiments, different light intensities and wavelengths were used. LAN exposure early in the dark phase at ZT15 caused increased glucose responses during the first 20 min after glucose infusion (p < 0.001), whereas LAN exposure at the end of the dark phase, at ZT21, caused increased insulin responses during the first 10 min (p < 0.01), indicating that LAN immediately induces glucose intolerance in rats. Subsequent experiments demonstrated that the effect of LAN was both intensity- and wavelength-dependent. White light of 50 and 150 lx induced greater glucose responses than 5 and 20 lx, whereas all intensities other than 5 lx reduced locomotor activity. Green light induced glucose intolerance, but red and blue light did not, suggesting the involvement of a specific retina-brain pathway. Together, these data show that exposure to LAN has acute adverse effects on glucose metabolism in a time-, intensity- and wavelength-dependent manner.


Additional sex comb-like (ASXL) proteins 1 and 2 play opposite roles in adipogenesis via reciprocal regulation of peroxisome proliferator-activated receptor {gamma}.
Our previous studies have suggested that the mammalian additional sex comb-like 1 protein functions as a coactivator or repressor of retinoic acid receptors in a cell-specific manner. Here, we investigated the roles of additional sex comb-like 1 proteins in regulating peroxisome proliferator-activated receptors (PPARs). In pulldown assays in vitro and in immunoprecipitation assays in vivo, ASXL1 and its paralog, ASXL2, interacted with PPARα and PPARγ. In 3T3-L1 preadipocyte cells, overexpression of ASXL1 inhibited the induction of PPARγ activity by rosiglitazone, as shown by transcription assays, and completely suppressed adipogenesis, as shown by Oil Red O staining. In contrast, overexpression of ASXL2 greatly enhanced rosiglitazone-induced PPARγ activity and enhanced adipogenesis. Deletion of the heterochromatin protein 1 (HP1)-binding domain from ASXL1 caused the mutant protein to enhance adipogenesis similarly to ASXL2, indicating that HP1 binding is required for the adipogenesis-suppressing activity of ASXL1. Adipocyte differentiation was associated with a gradual decrease in ASXL1 expression but did not affect ASXL2 expression. Knockdown of ASXL1 and ASXL2 had reciprocal effects on adipogenesis. In chromatin immunoprecipitation assays in 3T3-L1 cells, ASXL1 occupied the promoter of the PPARγ target gene aP2 together with HP1α and Lys-9-methylated histone H3, whereas ASXL2 occupied the aP2 promoter together with histone-lysine N-methyltransferase MLL1 and Lys-9-acetylated and Lys-4-methylated H3 histones. Finally, microarray analysis demonstrated that ASXL1 represses, whereas ASXL2 increases, the expression of adipogenic genes, most of which are PPARγ targets. These results suggest that members of the additional sex comb-like family provide complex regulation of adipogenesis via differential modulation of PPARγ activity.


Allogeneic adipose tissue-derived stem cells ELIXCYTE® in chronic kidney disease: A phase I study assessing safety and clinical feasibility.
The purpose of this phase I clinical trial is to assess the safety and tolerability of allogeneic adipose tissue-derived stem cells (ADSCs) among chronic kidney disease (CKD) patients. 12 eligible CKD patients with an estimated glomerular filtration rate (eGFR) of 15-44 ml/min/1.73 m2 received one dose of intravenous allogeneic ADSCs (ELIXCYTE® ), as 3 groups: 3 low dose (6.4 × 107 cells in total of 8 ml), 3 middle dose (19.2 × 107 cells in total of 24 ml) and 6 high dose (32.0 × 107 cells in total of 40 ml) of ELIXCYTE® and evaluated after 48 weeks. Primary endpoint was the safety profiles in terms of incidence of adverse events (AEs) and serious adverse event (SAE). Two subjects in high dose group experienced a total of 2 treatment-related AEs which are Grade 1 slow speech and Grade 1 bradyphrenia after the infusion. One subject in middle dose group experienced an SAE unlikely related to treatment, grade 2 proteinuria. No fatal AE was reported in this study. An increase in eGFR was observed in 7 out of 12 subjects (58%) at Week 24 and in 6 of 12 subjects (50%) by Week 48. By Week 24, an increase in eGFR by more than 20% among all CKD patients with baseline eGFR ≧ 30 ml/min/1.73 m2 as compared to only 2 subjects in baseline eGFR < 30 ml/min/1.73 m2 group. No significant reduction in proteinuria was noted among all subjects. This phase I trial demonstrated single-dose intravenous ELIXCYTE was well tolerated in moderate-to-severe CKD patients and its preliminary efficacy warrants future studies.


Multiple and Diverse vsp and vlp Sequences in Borrelia miyamotoi, a Hard Tick-Borne Zoonotic Pathogen.
Based on chromosome sequences, the human pathogen Borrelia miyamotoi phylogenetically clusters with species that cause relapsing fever. But atypically for relapsing fever agents, B. miyamotoi is transmitted not by soft ticks but by hard ticks, which also are vectors of Lyme disease Borrelia species. To further assess the relationships of B. miyamotoi to species that cause relapsing fever, I investigated extrachromosomal sequences of a North American strain with specific attention on plasmid-borne vsp and vlp genes, which are the underpinnings of antigenic variation during relapsing fever. For a hybrid approach to achieve assemblies that spanned more than one of the paralogous vsp and vlp genes, a database of short-reads from next-generation sequencing was supplemented with long-reads obtained with real-time DNA sequencing from single polymerase molecules. This yielded three contigs of 31, 16, and 11 kb, which each contained multiple and diverse sequences that were homologous to vsp and vlp genes of the relapsing fever agent B. hermsii. Two plasmid fragments had coding sequences for plasmid partition proteins that differed from each other from paralogous proteins for the megaplasmid and a small plasmid of B. miyamotoi. One of 4 vsp genes, vsp1, was present at two loci, one of which was downstream of a candiate prokaryotic promoter. A limited RNA-seq analysis of a population growing in the blood of mice indicated that of the 4 different vsp genes vsp1 was the one that was expressed. The findings indicate that B. miyamotoi has at least four types of plasmids, two or more of which bear vsp and vlp gene sequences that are as numerous and diverse as those of relapsing fever Borrelia. The database and insights from these findings provide a foundation for further investigations of the immune responses to this pathogen and of the capability of B. miyamotoi for antigenic variation.


Gene Family Evolution Reflects Adaptation to Soil Environmental Stressors in the Genome of the Collembolan Orchesella cincta.
Collembola (springtails) are detritivorous hexapods that inhabit the soil and its litter layer. The ecology of the springtail Orchesella cincta is extensively studied in the context of adaptation to anthropogenically disturbed areas. Here, we present a draft genome of an O. cincta reference strain with an estimated size of 286.8 Mbp, containing 20,249 genes. In total, 446 gene families are expanded and 1,169 gene families evolved specific to this lineage. Besides these gene families involved in general biological processes, we observe gene clusters participating in xenobiotic biotransformation. Furthermore, we identified 253 cases of horizontal gene transfer (HGT). Although the largest percentage of them originated from bacteria (37.5%), we observe an unusually high percentage (30.4%) of such genes of fungal origin. The majority of foreign genes are involved in carbohydrate metabolism and cellulose degradation. Moreover, some foreign genes (e.g., bacillopeptidases) expanded after HGT. We hypothesize that horizontally transferred genes could be advantageous for food processing in a soil environment that is full of decaying organic material. Finally, we identified several lineage-specific genes, expanded gene families, and horizontally transferred genes, associated with altered gene expression as a consequence of genetic adaptation to metal stress. This suggests that these genome features may be preadaptations allowing natural selection to act on. In conclusion, this genome study provides a solid foundation for further analysis of evolutionary mechanisms of adaptation to environmental stressors.


Nitranilic acid hexahydrate, a novel benchmark system of the Zundel cation in an intrinsically asymmetric environment: spectroscopic features and hydrogen bond dynamics characterised by experimental and theoretical methods.
Nitranilic acid (2,5-dihydroxy-3,6-dinitro-2,5-cyclohexadiene-1,4-dione) as a strong dibasic acid in acidic aqueous media creates the Zundel cation, H5O2(+). The structural unit in a crystal comprises (H5O2)2(+) (2,5-dihydroxy-3,6-dinitro-1,4-benzoquinonate)(2-) dihydrate where the Zundel cation reveals no symmetry, being an ideal case for studying proton dynamics and its stability. The Zundel cation and proton transfer dynamics are studied by variable-temperature X-ray diffraction, IR and solid-state NMR spectroscopy, and various quantum chemical methods, including periodic DFT calculations, ab initio molecular dynamics simulation, and quantization of nuclear motion along three fully coupled internal coordinates. The Zundel cation features a short H-bond with the O···O distance of 2.433(2) Å with an asymmetric placement of hydrogen. The proton potential is of a single well type and, due to the non-symmetric surroundings, of asymmetric shape. The formation of the Zundel cation is facilitated by the electronegative NO2 groups. The employed spectroscopic techniques supported by calculations confirm the presence of a short H-bond with a complex proton dynamics.


The effects of tocotrienols intake on obesity, blood pressure, inflammation, liver and glucose biomarkers: a meta-analysis of randomized controlled trials.
The objective of this study is to accomplish a systematic review and meta-analysis of all randomized controlled trials that dissected the influence of tocotrienol supplementation on various anthropometric and cardiometabolic indices in all individuals, irrespective of health condition. This research was carried out in line with the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) statement guidelines. 17 eligible articles were included in the final quantitative analysis. Current study revealed that tocotrienol consumption was not associated with CRP, WC, MDA, BMI, IL-6, HbA1C, ALT, AST, creatinine TNF-α, FPG, BW, DBP, and SBP. We did observe an overall increase in BW (SMD: 0.063 kg, 95% CI: -0.200, 0.327, p = 0.637) and DBP (SMD: 0.249 mmHg, 95% CI: 0.053, 0.446, p = 0.013). In addition, a significant reduction in SBP was observed (SMD: -0.616 mmHg, 95% CI: -1.123, -0.110, p = 0.017). In summary, our meta-analysis revealed that tocotrienol consumption was associated with increase in BW and DBP and decrease in SBP. Significant associations were not observed for other outcomes.


Contribution of different carbon sources to isoprene biosynthesis in poplar leaves.
This study was performed to test if alternative carbon sources besides recently photosynthetically fixed CO2 are used for isoprene formation in the leaves of young poplar (Populus x canescens) trees. In a 13CO2 atmosphere under steady state conditions, only about 75% of isoprene became 13C labeled within minutes. A considerable part of the unlabeled carbon may be derived from xylem transported carbohydrates, as may be shown by feeding leaves with [U-13C]Glc. As a consequence of this treatment approximately 8% to 10% of the carbon emitted as isoprene was 13C labeled. In order to identify further carbon sources, poplar leaves were depleted of leaf internal carbon pools and the carbon pools were refilled with 13C labeled carbon by exposure to 13CO2. Results from this treatment showed that about 30% of isoprene carbon became 13C labeled, clearly suggesting that, in addition to xylem transported carbon and CO2, leaf internal carbon pools, e.g. starch, are used for isoprene formation. This use was even increased when net assimilation was reduced, for example by abscisic acid application. The data provide clear evidence of a dynamic exchange of carbon between different cellular precursors for isoprene biosynthesis, and an increasing importance of these alternative carbon pools under conditions of limited photosynthesis. Feeding [1,2-13C]Glc and [3-13C]Glc to leaves via the xylem suggested that alternative carbon sources are probably derived from cytosolic pyruvate/phosphoenolpyruvate equivalents and incorporated into isoprene according to the predicted cleavage of the 3-C position of pyruvate during the initial step of the plastidic deoxyxylulose-5-phosphate pathway.


Design principles of regulatory networks: searching for the molecular algorithms of the cell.
A challenge in biology is to understand how complex molecular networks in the cell execute sophisticated regulatory functions. Here we explore the idea that there are common and general principles that link network structures to biological functions, principles that constrain the design solutions that evolution can converge upon for accomplishing a given cellular task. We describe approaches for classifying networks based on abstract architectures and functions, rather than on the specific molecular components of the networks. For any common regulatory task, can we define the space of all possible molecular solutions? Such inverse approaches might ultimately allow the assembly of a design table of core molecular algorithms that could serve as a guide for building synthetic networks and modulating disease networks.


Signaling from Rho to the actin cytoskeleton through protein kinases ROCK and LIM-kinase.
The actin cytoskeleton undergoes extensive remodeling during cell morphogenesis and motility. The small guanosine triphosphatase Rho regulates such remodeling, but the underlying mechanisms of this regulation remain unclear. Cofilin exhibits actin-depolymerizing activity that is inhibited as a result of its phosphorylation by LIM-kinase. Cofilin was phosphorylated in N1E-115 neuroblastoma cells during lysophosphatidic acid-induced, Rho-mediated neurite retraction. This phosphorylation was sensitive to Y-27632, a specific inhibitor of the Rho-associated kinase ROCK. ROCK, which is a downstream effector of Rho, did not phosphorylate cofilin directly but phosphorylated LIM-kinase, which in turn was activated to phosphorylate cofilin. Overexpression of LIM-kinase in HeLa cells induced the formation of actin stress fibers in a Y-27632-sensitive manner. These results indicate that phosphorylation of LIM-kinase by ROCK and consequently increased phosphorylation of cofilin by LIM-kinase contribute to Rho-induced reorganization of the actin cytoskeleton.


Genetic analyses and expression studies identified a novel mutation (W486C) as a molecular basis of congenital coagulation factor XII deficiency.
We analyzed the factor XII (FXII) gene of a patient with congenital FXII deficiency and identified a novel amino acid substitution (W486C) in the catalytic domain. The proband was an asymptomatic 49-year-old Japanese female with abnormal coagulation test, discovered by chance. The FXII activity and antigen level were both under 10%, suggesting a cross-reacting material-negative FXII deficiency. Sequence analysis of the proband's FXII gene revealed a homozygous nucleotide substitution G --> C in exon 12, resulting in the amino acid substitution W486C in the catalytic domain. We constructed the mutant FXII cDNA in an expression plasmid vector and transfected it into Chinese hamster ovary cells. The recombinant wild-type FXII antigen was detected in the culture medium by immunoprecipitation assay, but the mutant FXII (W486C) was not observed. On the other hand, both the wild-type FXII and W486C cell lysates contained FXII antigen and FXII mRNA, as estimated by western blotting and quantitative reverse transcriptase-polymerase chain reaction. These findings suggest that the W486C substitution of FXII impairs intracellular processing of the protein and/or transport system.


The transporter ABCB7 is a mediator of the phenotype of acquired refractory anemia with ring sideroblasts.
Refractory anemia with ring sideroblasts (RARS) is characterized by mitochondrial ferritin (FTMT) accumulation and markedly suppressed expression of the iron transporter ABCB7. To test the hypothesis that ABCB7 is a key mediator of ineffective erythropoiesis of RARS, we modulated its expression in hematopoietic cells. ABCB7 up and downregulation did not influence growth and survival of K562 cells. In normal bone marrow, ABCB7 downregulation reduced erythroid differentiation, growth and colony formation, and resulted in a gene expression pattern similar to that observed in intermediate RARS erythroblasts, and in the accumulation of FTMT. Importantly, forced ABCB7 expression restored erythroid colony growth and decreased FTMT expression level in RARS CD34+ marrow cells. Mutations in the SF3B1 gene, a core component of the RNA splicing machinery, were recently identified in a high proportion of patients with RARS and 11 of the 13 RARS patients in this study carried this mutation. Interestingly, ABCB7 exon usage differed between normal bone marrow and RARS, as well as within the RARS cohort. In addition, SF3B1 silencing resulted in downregulation of ABCB7 in K562 cells undergoing erythroid differentiation. Our findings support that ABCB7 is implicated in the phenotype of acquired RARS and suggest a relation between SF3B1 mutations and ABCB7 downregulation.


Nested partially latent class models for dependent binary data; estimating disease etiology.
The Pneumonia Etiology Research for Child Health (PERCH) study seeks to use modern measurement technology to infer the causes of pneumonia for which gold-standard evidence is unavailable. Based on case-control data, the article describes a latent variable model designed to infer the etiology distribution for the population of cases, and for an individual case given her measurements. We assume each observation is drawn from a mixture model for which each component represents one disease class. The model conisidered here addresses a major limitation of the traditional latent class approach by taking account of residual dependence among multivariate binary outcomes given disease class, hence reducing estimation bias, retaining efficiency and offering more valid inference. Such "local dependence" on each subject is induced in the model by nesting latent subclasses within each disease class. Measurement precision and covariation can be estimated using the control sample for whom the class is known. In a Bayesian framework, we use stick-breaking priors on the subclass indicators for model-averaged inference across different numbers of subclasses. Assessment of model fit and individual diagnosis are done using posterior samples drawn by Gibbs sampling. We demonstrate the utility of the method on simulated and on the motivating PERCH data.


Rural Families' Use of Multiple Child Care Arrangements from 6 to 58 Months and Children's Kindergarten Behavioral and Academic Outcomes.
Non-parental child care prior to kindergarten is a normative experience for the majority of children in the United States, with children commonly experiencing multiple arrangements, or more than one concurrent child care arrangement. The experience of multiple arrangements has predominantly been shown to be negatively related to young children's health and behavioral outcomes. The present study examined the use of multiple concurrent arrangements for children in the Family Life Project, a representative sample of families living in six high-poverty rural counties. Using the full sample of 1,292 children who were followed from six months to kindergarten, this study examined the associations between the number of child care arrangements averaged across six time points and children's behavioral and academic outcomes in kindergarten. After including a number of control variables, regression results suggested that a greater number of arrangements prior to kindergarten were related to higher levels of teacher-reported negative behaviors, but not positive behaviors, and letter-word decoding skills, but not mathematics skills, though effect sizes were small. Moderation analyses by child care type and quality were conducted, with no evidence emerging that findings varied by type or quality of care.


The effects of localized hypoventilation on ventilation/perfusion (V/Q) ratios and gas exchange in the dog lung.
1. Hypoventilation of one lobe of lung was studied in open-chest anaesthetized dogs. Lobar blood flow, pulmonary-artery pressure and gas exchange were measured, the latter from breath-by-breath analysis with a mass spectrometer. 2. Hypoventilation of the lobe by reducing the respiratory pump stroke led, at each step, to a reduction in blood flow to that lobe. The flow (Q) reduction was variable, but always less than the ventilation (VE) reduction, so that the V/Q ratio to the lobe was reduced. O2 tension and pH fell and CO2 tension rose in effluent blood. Thus V/Q regulation achieved by flow reduction varied between individuals and was of low gain. 3. Anatomical or series dead space (VD series) was reduced in proportion to ventilation. When VD series was less than the apparatus dead space, some gas exchange still took place. 4. Oxygen uptake (VO2) and CO2 output (VCO2) were reduced during hypoventilation. VCO2 fell more than VO2, so that the respiratory exchange ratio (R) was reduced. 5. Whether the deterioration in gas tensions in effluent blood during hypoventilation of the lobe was due to shunt of blood past unventilated alveoli, or to V/Q mismatching, was not resolved. 6. The plateau phase of the CO2-output curves at low tidal volumes was usually regular; thus either hypoventilation was uniform, or some ventilation units were totally closed.


A new highly adaptable design of shear-flow device for orientation of macromolecules for Linear Dichroism (LD) measurement.
This article presents a new design of flow-orientation device for the study of bio-macromolecules, including DNA and protein complexes, as well as aggregates such as amyloid fibrils and liposome membranes, using Linear Dichroism (LD) spectroscopy. The design provides a number of technical advantages that should make the device inexpensive to manufacture, easier to use and more reliable than existing techniques. The degree of orientation achieved is of the same order of magnitude as that of the commonly used concentric cylinders Couette flow cell, however, since the device exploits a set of flat strain-free quartz plates, a number of problems associated with refraction and birefringence of light are eliminated, increasing the sensitivity and accuracy of measurement. The device provides similar shear rates to those of the Couette cell but is superior in that the shear rate is constant across the gap. Other major advantages of the design is the possibility to change parts and vary sample volume and path length easily and at a low cost.


Daily Energy Expenditure and Its Relation to Health Care Costs in Patients Undergoing Ambulatory Electrocardiographic Monitoring.
Our increasingly sedentary lifestyle is associated with a heightened risk of obesity, diabetes, heart disease, and cardiovascular mortality. Using the recently developed heart rate index formula in 843 patients (mean ± SD age 62.3 ± 15.7 years) who underwent 24-hour ambulatory electrocardiographic (ECG) monitoring, we estimated average and peak daily energy expenditure, expressed as metabolic equivalents (METs), and related these data to subsequent hospital encounters and health care costs. In this cohort, estimated daily average and peak METs were 1.7 ± 0.7 and 5.5 ± 2.1, respectively. Patients who achieved daily bouts of peak energy expenditure ≥5 METs had fewer hospital encounters (p = 0.006) and median health care costs that were nearly 50% lower (p <0.001) than their counterparts who attained <5 METs. In patients whose body mass index was ≥30 kg/m2, there were significant differences in health care costs depending on whether they achieved <5 or ≥5 METs estimated by ambulatory ECG monitoring (p = 0.005). Interestingly, patients who achieved ≥5 METs had lower and no significant difference in their health care costs, regardless of their body mass index (p = 0.46). Patients with previous percutaneous coronary intervention who achieved ≥5 METs had lower health care costs (p = 0.044) and fewer hospital encounters (p = 0.004) than those who achieved <5 METs. In conclusion, average and peak daily energy expenditures estimated from ambulatory ECG monitoring may provide useful information regarding health care utilization in patients with and without previous percutaneous coronary intervention, irrespective of body habitus. Our findings are the first to link lower intensities of peak daily energy expenditure, estimated from ambulatory ECG monitoring, with increased health care utilization.


Pediatric Herpes Simplex Virus Encephalitis Complicated by N-Methyl-D-aspartate Receptor Antibody Encephalitis.
N-methyl-D-aspartate receptor antibodies (NMDAR-Abs) can contribute to neurological relapse after herpes simplex virus encephalitis (HSE). We describe a child with NMDAR-Ab encephalitis after HSE, which was recognized and treated early. We discuss the case in the context of existing reports, and we propose a modified immunotherapy strategy to minimize risk of viral reactivation.


Highly effective degradation of sodium dodecylbenzene sulphonate and synthetic greywater by Fenton-like reaction over zerovalent iron-based catalyst.
There is an increasing interest to recycle greywater for meeting non-portable water demand. However, linear alkylbenzene sulphonates (a form of anionic surfactants) that are commonly found in greywater are less biodegradable at moderate to high concentrations. A fenton-like system is a relatively economic advanced oxidation process that can potentially be used for surfactant degradation in greywater treatment. This study investigated the feasibility of zerovalent iron (ZVI)-mediated Fenton's oxidation of sodium dodecylbenzene sulphonate (SDBS) using Fe0/H2O2 and Fe2+/Fe0/H2O2 systems under a range of operating conditions. For the Fe0/H2O2 binary system, the initial pH value and Fe0 dosage played important roles in final degradation efficiency. For the Fe2+/Fe0/H2O2 ternary systems, a small amount of Fe2+ (0.5-1.7 mM) contributed a synergistic effect to promote iron recycling and SDBS degradation. Approximately, 90% of SDBS mineralization efficiency was accomplished within 15 min at a pH range from 3.0 to 6.5, using 18 mM Fe0 and 15 mM H2O2. However, the removal kinetics was rate-limited by Fe2+ dissolution from the ZVI surfaces. The Fenton-like process of the Fe2+/Fe0/H2O2 ternary system also presents a promising treatment method for synthetic greywater, in which 90% TOC removal was achieved within the first 10 min; 78% COD and 91% BOD5 were achieved after 120 min of reaction.


Activating mutations in the N- and K-ras oncogenes differentially affect the growth properties of the IL-6-dependent myeloma cell line ANBL6.
Although the underlying genetic defect in multiple myeloma is unknown, activating mutations in the N- and K-ras oncogenes are common. Recent studies have suggested that ras mutations are associated with disease progression. We have introduced an activated N-ras12, N-ras61, or K-ras12 cDNA into the interleukin 6 (IL-6)-dependent multiple myeloma cell line ANBL6 to determine the effect of N- and K-ras on the growth/death properties of ANBL6. All three transduced cell populations demonstrate a growth advantage over the parent ANBL6 when propagated on normal human bone marrow stromal cells. In the absence of bone marrow stromal cells, augmentation of growth was observed in all three mutant ras-expressing populations at optimal and suboptimal concentrations of IL-6. Furthermore, in the absence of IL-6, all mutant ras populations demonstrated an augmentation in DNA synthesis when compared to the parent ANBL6. However, growth of the K-ras12 population in the absence of IL-6 was significantly inhibited when compared to the mutant N-ras populations. This could be explained by the observation that in the absence of IL-6, N-ras12 and N-ras61 suppress apoptosis, whereas K-ras12 does not. We also found that mutant ras expression could result in early protection from glucocorticoid-induced apoptosis similar to that observed by the addition of IL-6. However, the combination of mutant ras and IL-6 could completely block the glucocorticoid induction of apoptosis in long-term cultures. These data suggest that mutations in different ras family members may have similar or distinct effects on myeloma tumor growth and death and may alter the response to glucocorticoid treatment.


Fidelity to the healing relationship: a medical student's challenge to contemporary bioethics and prescription for medical practice.
As a medical student, I observed that different physicians had strikingly different attitudes and approaches when caring for patients. The care of one patient in particular continues to challenge my understanding of illness and moral responsibility in the practice of medicine. In this paper, I illustrate the care of this patient in order to evaluate the dominant ethics I was taught in medical school, in theory and in practice, and argue neither principlism nor the ethics of care fully captures the moral responsibility of physicians. Emphasising fidelity to the healing relationship, a core principle derived from Pellegrino's virtue theory, I conclude that this approach to clinical ethics fully explains physician responsibility. Pellegrino deconstructs the practice of medicine to clarify the moral event within the clinical encounter and offers a sufficiently useful and justified approach to patient care.


Limited proteolysis of human histone deacetylase 1.
Histone deacetylase (HDAC) proteins are associated with cell proliferation, differentiation, apoptosis, and cancer. Specifically, HDAC1 is linked with cell growth, a hallmark of cancer formation. HDAC1 is a phosphoprotein and phosphorylation at S421 and S423 promotes HDAC1 enzymatic activity and protein association. While single and double point mutants of HDAC1 at S421 and S423 appear functionally similar, the evidence suggests that HDAC1 is phosphorylated simultaneously at both S421 and S423 in vivo. Additional experiments are necessary to probe the role of double phosphorylation of HDAC1 at S421 and S423. To characterize HDAC1 phosphorylation at S421 and S423, limited proteolysis of HDAC1 was performed for the first time. HDAC1 degraded without production of discrete fragments. By performing concentration-dependent proteolysis, HDAC1 double point mutants with disrupted phosphorylation at S421 and S423 displayed different trypsin sensitivities compared to wild type HDAC1. Unexpectedly, HDAC1 single point mutants with disrupted phosphorylation at either S421 or S423 demonstrated protease sensitivity similar to the wild type HDAC1. Concentration-dependent proteolysis experiments provide evidence that phosphorylation of S421 and S423 individually contribute to HDAC1 function. In addition, the limited proteolysis experiments support a model where associated proteins promote HDAC1 enzymatic activity, reinforcing the importance of protein interactions in HDAC1 structure and function. Finally, because HDAC1 does not display distinct regions of protease sensitivity, the proteolysis studies suggest that HDAC1 comprises inter-related structural regions.


Substitutions of Ser83Leu in GyrA and Ser80Leu in ParC Associated with Quinolone Resistance in Acinetobacter pittii.
To investigate the prevalence and the mechanism of quinolone-resistant Acinetobacter pittii, 634 Acinetobacter calcoaceticus-Acinetobacter baumannii complex isolates were collected throughout Zhejiang Province. Identification of isolates was conducted by matrix-assisted laser desorption ionization/time of flight mass spectrometry (MALDI-TOF MS), blaOXA-51-like gene, and partial RNA polymerase β-subunit (rpoB) amplification. Twenty-seven isolates of A. pittii were identified. Among the 634 isolates, A. baumannii, A. pittii, Acinetobacter nosocomialis, and A. calcoaceticus counted for 87.22%, 4.26%, 8.20%, and 0.32%, respectively. Antimicrobial susceptibility of nalidixic acid, ofloxacin, enoxacin, ciprofloxacin, lomefloxacin, levofloxacin, sparfloxacin, moxifloxacin, and gatifloxacin for 27 A. pittii were determined by the agar dilution method. Detection of quinolone-resistant determining regions of gyrA, gyrB, parC, and parE was performed for the A. pittii isolates. In addition, plasmid-mediated quinolone resistance (PMQR) determinants (qnrA, qnrB, qnrS, qnrC, qnrD, aac(6')-Ib-cr, qepA, oqxA, and oqxB) were investigated. All the 27 isolates demonstrated a higher minimum inhibitory concentration (MIC) to old quinolones than the new fluoroquinolones. No mutation in gyrA, gyrB, parC, or parE was detected in 20 ciprofloxacin-susceptible isolates. Seven ciprofloxacin-resistant A. pittii were identified with a Ser83Leu mutation in GyrA. Among them, six isolates with simultaneous Ser83Leu amino acid substitution in GyrA and Ser80Leu in ParC displayed higher MIC values against ciprofloxacin. Additionally, three were identified with a Met370Ile substitution in ParE, and two were detected with a Tyr317His mutation in ParE, which were reported for the first time. No PMQR determinants were identified in the 27 A. pittii isolates. In conclusion, mutations in chromosome play a major role in quinolone resistance in A. pittii, while resistance mechanisms mediated by plasmid have not been found. Ser83Leu substitution in GyrA and Ser80Leu substitution in ParC are associated with quinolone resistance in A. pittii. Whether Met370Ile and Tyr317His substitutions in ParE play a minor role requires further investigation.


Biting patterns of malaria vectors of the lower Shire valley, southern Malawi.
Assessing the biting behaviour of malaria vectors plays an integral role in understanding the dynamics of malaria transmission in a region. Biting times and preference for biting indoors or outdoors varies among mosquito species and across regions. These behaviours may also change over time in response to vector control measures such as long-lasting insecticidal nets (LLINs). Data on these parameters can provide the sites and times at which different interventions would be effective for vector control. This study assessed the biting patterns of malaria vectors in Chikwawa district, southern Malawi. The study was conducted during the dry and wet seasons in 2016 and 2017, respectively. In each season, mosquitoes were collected indoors and outdoors for 24 nights in six houses per night using the human landing catch. Volunteers were organized into six teams of two individuals, whereby three teams collected mosquitoes indoors and the other three collected mosquitoes outdoors each night, and the teams were rotated among twelve houses. All data were analyzed using Poisson log-linear models. The most abundant species were Anopheles gambiae sensu lato (primarily An. arabiensis) and An. funestus s.l. (exclusively An. funestus s.s.). During the dry season, the biting activity of An. gambiaes.l. was constant outdoors across the categorized hours (18:00 h to 08:45 h), but highest in the late evening hours (21:00 h to 23:45 h) during the wet season. The biting activity of An. funestus s.l. was highest in the late evening hours (21:00 h to 23:45 h) during the dry season and in the late night hours (03:00 h to 05:45 h) during the wet season. Whereas the number of An. funestuss.l. biting was constant (P = 0.662) in both seasons, that of An. gambiaes.l. was higher during the wet season than in the dry season (P = 0.001). Anopheles gambiae s.l. was more likely to bite outdoors than indoors in both seasons. During the wet season, An. funestus s.l. was more likely to bite indoors than outdoors but during the dry season, the bites were similar both indoors and outdoors. The biting activity that occurred in the early and late evening hours, both indoors and outdoors coincides with the times at which individuals may still be awake and physically active, and therefore unprotected by LLINs. Additionally, a substantial number of anopheline bites occurred outdoors. These findings imply that LLINs would only provide partial protection from malaria vectors, which would affect malaria transmission in this area. Therefore, protection against bites by malaria mosquitoes in the early and late evening hours is essential and can be achieved by designing interventions that reduce vector-host contacts during this period.


Harmine suppresses the proliferation of pancreatic cancer cells and sensitizes pancreatic cancer to gemcitabine treatment.
Pancreatic carcinoma is one of the most deadliest types of cancer, and relatively insensitive to the currently available chemotherapy. Thus, the discovery of novel therapeutic agents to prolong the survival times of patients with pancreatic cancer is urgently required. Cell proliferation was assessed using the sulforhodamine B and cell clone formation assay, apoptosis was analyzed through Annexin V/PI staining, analysis of cell cycle distribution was determined by PI staining, and the expression of proteins was detected via Western blotting. Our data showed that harmine exerted an anti-proliferative effect and cell cycle arrest at G2/M in pancreatic cancer cells. Meanwhile, harmine plus gemcitabine showed strong synergy in inhibiting the proliferation of pancreatic cancer cells. Furthermore, harmine induced apoptosis and enhanced the gemcitabine-induced apoptosis in pancreatic cancer cells. The AKT/mTOR pathway is involved in mechanisms of gemcitabine resistance in pancreatic cancer cells, our data demonstrated that harmine plus gemcitabine significantly suppressed the AKT/mTOR signaling pathway. Harmine may be a potential candidate for the treatment of pancreatic cancer. Morever, the combination of harmine with gemcitabine appears to be an attractive option for the treatment of patients with pancreatic cancer.


Evaluation of ozone for preventing fungal influenced corrosion of reinforced concrete bridges over the River Nile, Egypt.
Fungal influenced corrosion (FIC) of some corroded sites in three selected bridges [Embaba bridge (E-bridge), Kasr al-Nile-bridge (K-bridge) and University bridge (U-bridge)] located over the River Nile in Egypt were investigated. Six fungal species, belong to 12 fungal genera, were isolated from the corroded reinforced concrete of the three tested bridges. Fourier transform infrared spectroscopy (FTIR) was screened for the most dominant fungal species (Fusarium oxysporium) which showed in all tested bridges that indicated the presence of amine group accompanied with polysaccharides contents. FIC of the most deteriorated bridge (K-bridge) was documented with FTIR. The association of fungal spores with corrosion products was recorded with scanning electron microscope (SEM). Evaluation of ozone for preventing FIC of the K-bridge was carried out by recording the corrosion rate and the corresponding inhibition efficiency (IE%). No mycelial growth with 100% IE was observed at 3 ppm ozone concentration after 120 min exposure time. With longer duration of ozone exposure, the membrane permeability of F. oxysporium was compromised as indicated by protein and nucleic acid leakages accompanied with lipid and tryptophan oxidation. The total intracellular and extracellular proteins of F. oxysporium were run on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) indicated the increasing of the supernatant protein on the expense of the cellular protein bands with extending ozone exposure time (0-80 min).


Fibronectin and serum amyloid P component stimulate C3b- and C3bi-mediated phagocytosis in cultured human monocytes.
Fibronectin (FN) and serum amyloid P component (SAP) markedly enhance phagocytosis mediated by the C3b and C3bi receptors of cultured human monocytes but not of granulocytes. (The C3b and C3bi receptors of granulocytes can be activated by treatment of these phagocytes with PMA.) Activation of monocyte C3 receptors by FN is developmentally regulated: Freshly explanted monocytes respond to FN with a small increase in C3 receptor-mediated phagocytosis while monocytes matured in culture exhibit a much greater response. The mechanism of action of FN on C3 receptors of cultured monocytes is unique in two respects. First, while substrate-bound FN or SAP activate monocyte C3 receptors, soluble FN does not. Second, stimulation of the basal surface of monocyte plasma membranes by substrate-bound FN activates C3b and C3bi receptors on the apical surface of the plasma membrane, i.e., at sites remote from the segments of membrane in contact with the FN or SAP.


The partial amino acid sequence of the extracellular beta-lactamase I of Bacillus cereus 569/H.
The chemical structure of the extracellular beta-lactamase I of Bacillus cereus 569/H was investigated. Three electrophoretically homogenous charge variants of this enzyme were isolated and amino acid analysis of each revealed no significant differences. However, a degree of N-terminal heterogeneity was found by direct end-group modification of the protein and also on alignment of peptides from tryptic and chymotryptic digestion. The N-terminal heterogeneity observed was great enough to explain the production of the beta-lactamase I isoenzymes which are probably produced by postsynthesis modification of a single gene product. Over 80% of the amino acid sequence of beta-lactamase I was determined by the detailed analysis of peptides derived from tryptic, chymotryptic and thermolytic digests. Five polypeptide fragments were constructed from these data and aligned by comparison with the known amino acid sequences of the penicillinases produced by Bacillus licheniformis and Staphylococcus aureus (Ambler & Meadway, 1969). About 60% of the proposed sequence was identical with that of B. licheniformis penicillinase, whereas the S. aureus enzyme had only about 40% of its residues in common with beta-lactamase I. These results are discussed with reference to the possible evolutionary relationships existing between known beta-lactamases. Detailed evidence for the amino acid sequence proposed has been deposited as Supplementary Publication SUP 50044 (27 pages) at the British Library (Lending Division), Boston Spa, Wetherby, W. Yorks. LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1975), 145, 5.


Review of the buccal-attaching fish parasite genus Glossobius Schioedte & Meinert, 1883 (Crustacea: Isopoda: Cymothoidae).
Two species of Glossobius Schioedte & Meinert, 1883 are known from Australia: Glossobius anctus Bruce & Bowman, 1989 and Glossobius impressus (Say, 1818), the latter recorded here for the first time from Australia and southern Africa. Glossobius ogasawarensis Nunomura, 1994 is here placed in synonymy with Glossobius auritus Bovallius, 1885; whereas Glossobius crassa (Dana, 1853) is removed from synonymy with G. auritus and placed into nomen dubium. Glossobius arimae Nunomura, 2001 is transferred to the genus Ceratothoa Dana, 1852. A key to the species of Glossobius is presented.


Quantum dynamics of the vibrations of helium bound to the nanosurface of a large planar organic molecule: phthalocyanine . He van der Waals complex.
We report rigorous quantum three-dimensional calculations of highly excited intermolecular vibrational states of the van der Waals (vdW) complex phthalocyanine.He (Pc.He). The Pc molecule was treated as rigid and the intermolecular potential energy surface (IPES) was represented as a sum of atom-atom Lennard-Jones pair potentials. The IPES has four equivalent global minima on the diagonals of the square-shaped Pc, inside its five-membered rings, and four slightly shallower local minima between them, creating a distinctive corrugation pattern of the molecular nanosurface. The vdW vibrational states analyzed in this work extend to about two-thirds of the well depth of the IPES. For the assignment of the in-plane (xy) vdW vibrational excitations it was necessary to resort to two sets of quantum numbers, the Cartesian quantum numbers [nu(x), nu(y)] and the quantum numbers (v, l) of the 2D isotropic oscillator, depending on the nodal structure and the symmetry of the wave functions. The delocalization of the He atom parallel to the molecular surface is large already in the ground vdW state. It increases rapidly with the number of quanta in the in-plane vdW vibrations, with the maximum root-mean-square amplitudes Deltax and Deltay of about 7 au at the excitation energies around 40 cm(-1). The wave functions of the highly excited states tend to be delocalized over the entire nanosurface and often have a square shape, reflecting that of the substrate.


A picture paints a thousand words: Heart drawings reflect acute distress and illness perception and predict posttraumatic stress symptoms after acute myocardial infarction.
The aim of this study was to examine whether heart drawings of patients with acute myocardial infarction reflect acute distress symptoms and negative illness beliefs and predict posttraumatic stress symptoms 3 months post-myocardial infarction. In total, 84 patients aged over 18 years drew pictures of their heart. The larger the area drawn as damaged, the greater were the levels of acute distress (r = 0.36; p < 0.05), negative illness perceptions (r = 0.42, p < 0.05), and posttraumatic stress symptoms (r = 0.54, p < 0.01). Pain drawings may offer a tool to identify maladaptive cognitions and thus patients at risk of posttraumatic stress disorder.


Incremental slow feature analysis: adaptive low-complexity slow feature updating from high-dimensional input streams.
We introduce here an incremental version of slow feature analysis (IncSFA), combining candid covariance-free incremental principal components analysis (CCIPCA) and covariance-free incremental minor components analysis (CIMCA). IncSFA's feature updating complexity is linear with respect to the input dimensionality, while batch SFA's (BSFA) updating complexity is cubic. IncSFA does not need to store, or even compute, any covariance matrices. The drawback to IncSFA is data efficiency: it does not use each data point as effectively as BSFA. But IncSFA allows SFA to be tractably applied, with just a few parameters, directly on high-dimensional input streams (e.g., visual input of an autonomous agent), while BSFA has to resort to hierarchical receptive-field-based architectures when the input dimension is too high. Further, IncSFA's updates have simple Hebbian and anti-Hebbian forms, extending the biological plausibility of SFA. Experimental results show IncSFA learns the same set of features as BSFA and can handle a few cases where BSFA fails.


Isolation and characterization of endophytic plant growth-promoting bacteria from date palm tree (Phoenix dactylifera L.) and their potential role in salinity tolerance.
Endophytic bacteria were isolated from date palm (Phoenix dactylifera L.) seedling roots, characterized and tested for their ability to help plants grow under saline conditions. Molecular characterization showed that the majority of these strains belonged to the genera Bacillus and Enterobacter and had different degrees of resistance to various antibiotics. Some of these strains were able to produce the enzyme 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase and the plant growth regulatory hormone indole-3-acetic acid (IAA). Some strains were also able to chelate ferric iron (Fe(3+)) and solubilize potassium (K(+)), phosphorus (PO 4 (3-) ) and zinc (Zn(2+)), and produce ammonia. The results also showed that ACC deaminase activity and IAA production was slightly increased in some strains in response to an increase in NaCl concentration in the growth media. Consistent with these results, selected strains such as PD-R6 (Paenibacillus xylanexedens) and PD-P6 (Enterobacter cloacae) were able to enhance canola root elongation when grown under normal and saline conditions as demonstrated by a gnotobiotic root elongation assay. These results suggest that the isolated and characterized endophytic bacteria can alter ethylene and IAA levels and also facilitate nutrient uptake in roots and therefore have the potential role to promote the growth and development of date palm trees growing under salinity stress.