Alterations in Notch signalling in skeletal muscles from mdx and dko dystrophic mice and patients with Duchenne muscular dystrophy.
New Findings What is the central question of this study? The Notch signalling pathway plays an important role in muscle regeneration, and activation of the pathway has been shown to enhance muscle regeneration in aged mice. It is unknown whether Notch activation will have a similarly beneficial effect on muscle regeneration in the context of Duchenne muscular dystrophy (DMD). What is the main finding and its importance? Although expression of Notch signalling components is altered in both mouse models of DMD and in human DMD patients, activation of the Notch signalling pathway does not confer any functional benefit on muscles from dystrophic mice, suggesting that other signalling pathways may be more fruitful targets for manipulation in treating DMD. Abstract In Duchenne muscular dystrophy (DMD), muscle damage and impaired regeneration lead to progressive muscle wasting, weakness and premature death. The Notch signalling pathway represents a central regulator of gene expression and is critical for cellular proliferation, differentiation and apoptotic signalling during all stages of embryonic muscle development. Notch activation improves muscle regeneration in aged mice, but its potential to restore regeneration and function in muscular dystrophy is unknown. We performed a comprehensive examination of several genes involved in Notch signalling in muscles from dystrophin-deficient mdx and dko (utrophin- and dystrophin-null) mice and DMD patients. A reduction of Notch1 and Hes1 mRNA in tibialis anterior muscles of dko mice and quadriceps muscles of DMD patients and a reduction of Hes1 mRNA in the diaphragm of the mdx mice were observed, with other targets being inconsistent across species. Activation and inhibition of Notch signalling, followed by measures of muscle regeneration and function, were performed in the mouse models of DMD. Notch activation had no effect on functional regeneration in C57BL/10, mdx or dko mice. Notch inhibition significantly depressed the frequency-force relationship in regenerating muscles of C57BL/10 and mdx mice after injury, indicating reduced force at each stimulation frequency, but enhanced the frequency-force relationship in muscles from dko mice. We conclude that while Notch inhibition produces slight functional defects in dystrophic muscle, Notch activation does not significantly improve muscle regeneration in murine models of muscular dystrophy. Furthermore, the inconsistent expression of Notch targets between murine models and DMD patients suggests caution when making interspecies comparisons.


Gain-of-function mutation of a voltage-gated sodium channel NaV1.7 associated with peripheral pain and impaired limb development.
Dominant mutations in voltage-gated sodium channel NaV1.7 cause inherited erythromelalgia, a debilitating pain disorder characterized by severe burning pain and redness of the distal extremities. NaV1.7 is preferentially expressed within peripheral sensory and sympathetic neurons. Here, we describe a novel NaV1.7 mutation in an 11-year-old male with underdevelopment of the limbs, recurrent attacks of burning pain with erythema, and swelling in his feet and hands. Frequency and duration of the episodes gradually increased with age, and relief by cooling became less effective. The patient's sister had short stature and reported similar complaints of erythema and burning pain, but with less intensity. Genetic analysis revealed a novel missense mutation in NaV1.7 (2567G>C; p.Gly856Arg) in both siblings. The G856R mutation, located within the DII/S4-S5 linker of the channel, substitutes a highly conserved non-polar glycine by a positively charged arginine. Voltage-clamp analysis of G856R currents revealed that the mutation hyperpolarized (-11.2 mV) voltage dependence of activation and slowed deactivation but did not affect fast inactivation, compared with wild-type channels. A mutation of Gly-856 to aspartic acid was previously found in a family with limb pain and limb underdevelopment, and its functional assessment showed hyperpolarized activation, depolarized fast inactivation, and increased ramp current. Structural modeling using the Rosetta computational modeling suite provided structural clues to the divergent effects of the substitution of Gly-856 by arginine and aspartic acid. Although the proexcitatory changes in gating properties of G856R contribute to the pathophysiology of inherited erythromelalgia, the link to limb underdevelopment is not well understood.


The Kip3-like kinesin KipB moves along microtubules and determines spindle position during synchronized mitoses in Aspergillus nidulans hyphae.
Kinesins are motor proteins which are classified into 11 different families. We identified 11 kinesin-like proteins in the genome of the filamentous fungus Aspergillus nidulans. Relatedness analyses based on the motor domains grouped them into nine families. In this paper, we characterize KipB as a member of the Kip3 family of microtubule depolymerases. The closest homologues of KipB are Saccharomyces cerevisiae Kip3 and Schizosaccharomyces pombe Klp5 and Klp6, but sequence similarities outside the motor domain are very low. A disruption of kipB demonstrated that it is not essential for vegetative growth. kipB mutant strains were resistant to high concentrations of the microtubule-destabilizing drug benomyl, suggesting that KipB destabilizes microtubules. kipB mutations caused a failure of spindle positioning in the cell, a delay in mitotic progression, an increased number of bent mitotic spindles, and a decrease in the depolymerization of cytoplasmic microtubules during interphase and mitosis. Meiosis and ascospore formation were not affected. Disruption of the kipB gene was synthetically lethal in combination with the temperature-sensitive mitotic kinesin motor mutation bimC4, suggesting an important but redundant role of KipB in mitosis. KipB localized to cytoplasmic, astral, and mitotic microtubules in a discontinuous pattern, and spots of green fluorescent protein moved along microtubules toward the plus ends.


Indocyanine Green Angiography as the Principal Design and Perfusion Assessment Tool for the Supraclavicular Artery Island Flap in Head and Neck Reconstruction.
A consecutive case series of supraclavicular artery island flaps was designed using indocyanine green angiography (IcG-A) in head and neck reconstruction to demonstrate its utilization in supraclavicular artery island flap (SCAIF) head and neck reconstruction. IcG-A was used consecutively between April 2014 and July 2015 to evaluate its use in flap design, inset, and intraoperative decision-making in five patients undergoing head and neck reconstruction. Six SCAIFs were harvested in five patients where IcG-A was used as the primary tool for flap design by visually mapping the supraclavicular artery under fluorescence. Each flap was harvested around the mapped course of the artery. Confirmatory Doppler was present in each flap raised with this technique. In all five patients, IcG-A was used to assess flap perfusion after inset. This case series demonstrates IcG-A as another tool for SCAIF design in head and neck reconstruction. The technology provides direct visualization of the pedicle before harvest. It can also be used as an intraoperative tool to visualize the blood supply once the flap is rotated to assess flap perfusion and detect areas that may be compromised, thereby improving flap survival.


Focusing light with a reflection photon sieve.
An advanced type of diffractive optical element is presented that combines the concept of the photon sieve with an off-axis, off-normal incidence reflection geometry. Compared to transmission optical elements, the signal-to-background ratio is significantly increased by separating the first from other diffraction orders without drastically reducing the size of the smallest diffractive element. The reflection photon sieve produces sharp foci at maximum contrast and offers the advantages of effective heat dissipation and a large working space above the focal plane. Experimental results for a device working at a photon energy of 100 eV are presented and compared to theory.


Complexin-1 and Foxp1 Expression Changes Are Novel Brain Effects of Alpha-Synuclein Pathology.
As the second most frequent neurodegenerative disorder of the aging population, Parkinson's disease (PD) is characterized by progressive deficits in spontaneous movement, atrophy of dopaminergic midbrain neurons and aggregation of the protein alpha-synuclein (SNCA). To elucidate molecular events before irreversible cell death, we studied synucleinopathy-induced expression changes in mouse brain and identified 49 midbrain/brainstem-specific transcriptional dysregulations. In particular complexin-1 (Cplx1), Rabl2a and 14-3-3epsilon (Ywhae) downregulation, as well as upregulation of the midbrain-specific factor forkhead box P1 (Foxp1) and of Rabgef1, were interesting as early mRNA level effects of alpha-synuclein triggered pathology. The protein levels of complexin-1 were elevated in midbrain/brainstem tissue of mice with A53T-SNCA overexpression and of mice with SNCA-knockout. The response of CPLX1 and Foxp1 levels to SNCA deficiency supports the notion that these factors are regulated by altered physiological function of alpha-synuclein. Thus, their analysis might be useful in PD stages before the advent of Lewy pathology. Because both alpha-synuclein and complexin-1 modulate vesicle release, our findings support presynaptic dysfunction as an early event in PD pathology.


Effects of dance on depression, physical function, and disability in underserved adults.
This study documented the feasibility and immediate effects of a dance intervention two times per week for 12 weeks on depression, physical function, and disability in older, underserved adults. The one-group, pretest-posttest study had a convenience sample of 40 participants recruited from a federally subsidized apartment complex located in an economically depressed, inner-city neighborhood. Depression, physical function, and disability were measured at baseline and 12 weeks. Average age was 63 years (SD = 7.9), 92% were female, and 75% were African American. At baseline, participants reported increased depression (M = 20.0, SD = 12.4), decreased physical function (M = 56.6, SD = 10.9), and increased disability limitations (M = 65.7, SD = 14.9). At posttest, paired t tests showed that the dance intervention significantly decreased depression, t = 6.11, p < .001, and disability, t = -2.70, p = .014, and significantly increased physical function, t = -2.74, p = .013. The results indicate that the 12-week dance intervention may be an effective adjunct therapy to improve depression, disability, and physical function in underserved adults.


The heparin binding PECAM-1 adhesion molecule is expressed by CD34+ hematopoietic precursor cells with early myeloid and B-lymphoid cell phenotypes.
The platelet-endothelial cell adhesion molecule-1 (PE-CAM-1), defined by the CD31 monoclonal antibody (MoAb), was initially described as a cell-cell adhesion molecule mediating both homotypic and heterotypic adhesion. In this report, we show that enriched CD34+ human hematopoietic progenitor cell populations, containing early myeloid, erythroid, and multipotential progenitor cells, are CD31+. Analyses of CD34+ cell lines representing early myeloid, multipotential, and pre-pre-B-lymphoid progenitors indicate that precursors of both myeloid and B-lymphoid cells express PECAM-1 at high levels. Three-color flow-cytometric analyses also show that normal human bone marrow CD31+ CD34+ subsets coexpress myeloid (CD33) or B-lymphoid (CD19, CD10) markers. Except for the monocytic cell line, U937, all CD34- cell lines tested, which represent more mature stages of the myeloid, erythroid, and lymphoid lineages, expressed substantially lower or negligible levels of PECAM-1. Western blotting studies indicated that the CD31 MoAb, JC/70A, detected molecules in the 120- to 140-kD molecular weight range on the monocytic CD34- CD33+ CD31+ cell line, U937; on the CD34+ CD31+ CD33+ CD19- multipotential/lymphomyeloid precursor cell lines, KG1 and KG1B; on the CD34+ CD31+ CD19+ CD10+ CD33- precursor pre-pre-B-cell line, MIK-ALL; and on a CD34(+)-enriched precursor cell population from normal human bone marrow. A single molecular weight species was generally observed with enriched membrane preparations, whereas two PECAM-1 molecules were present in whole-cell lysates of cell lines and the CD34+ bone marrow cell subset. Preliminary studies show that a proportion of the PECAM-1 molecules on the lymphomyeloid/multipotential progenitor cell line, KG1, and on the monocytic cell line, U937, binds to heparin-sepharose. A soluble form of PECAM-1 also binds heparin-sepharose. The high level of expression of PECAM-1 on CD34+ cells suggests that this glycoprotein may function as a heterotypic adhesion molecule, possibly mediating multipotential, myeloid, and early-B-lymphoid precursor cell interactions with stromal cells and extracellular matrix molecules via heparan sulfate proteoglycans. It may also act as a homotypic adhesion molecule by interacting with PECAM-1 on bone marrow stromal macrophage-like cells and endothelial cells or on endothelial cells during stem/progenitor cell migration. Thus, this molecule has the potential importance of directing both lineage commitment and trafficking of early hematopoietic progenitor cells.


Global analysis of gene expression in invasion by a lung cancer model.
Metastasis is a complicated multistep process that involves interactions between cancer cells and their surrounding microenvironments. Previously, we have established a series of lung adenocarcinoma cell lines with varying degrees of invasiveness. Tracheal graft assay confirmed that cell lines with higher in vitro invasiveness had greater in vivo invasive potential. In this study, we used these model cell lines to identify invasion-associated genes using cDNA microarray with colorimetric detection. A more invasive subline, CL 1-5-F 4, derived from metastatic lung tumor of severe combined immunodeficient mice inoculated with CL 1-5 cells, was combined with CL 1-0, CL 1-1, and CL 1-5 in cDNA microarray screening. cDNA microarray membranes, each containing 9600 nonredundant expressed sequence tag clones, were used to identify differentially expressed genes in these cell lines. For statistical analysis, self-organizing map algorithm was performed to identify the expression patterns. Positive correlation between gene expression levels and cell line invasiveness was found in 2.9% of the 9600 putative genes. On the other hand, negative correlation was found in 3.3% of the genes. The trends of expression of some of the genes were also confirmed by Northern hybridization and flow cytometry. Our data demonstrated that genes related to cell adhesion, motility, angiogenesis, signal transduction, and some other expressed sequence tag genes may play significant roles in the metastasis process. These results substantiate the model system with which one can identify invasion-associated genes by using cDNA microarray and cancer cell lines of different invasiveness. This technique may allow us to explore complex interactions between multiple genes that orchestrate the process of cancer metastasis.


Coding for quality measurement: the relationship between hospital structural characteristics and coding accuracy from the perspective of quality measurement.
This study examines the relationship between hospital structural characteristics and coding accuracy from the perspective of quality measurement. To measure coding accuracy for quality measurement, the study utilizes the "present on admission" indicator, a data element in the New York state hospital administrative database. This data element is used by hospitals across New York state to indicate if a particular secondary diagnosis is "present on admission," "not present on admission," or "uncertain." Since the accurate distinction between comorbidities (present at admission) and complications (not present at admission,) is critical for risk adjustment in comparative hospital quality reports, this study uses the occurrence of the value "uncertain" in the "present on admission" indicator as the primary measure of coding accuracy. A lower occurrence of the value "uncertain" is considered to be reflective of better coding accuracy. Moreover, since coding accuracy of the "present on admission" indicator links back to the accuracy of physician documentation, a focus on the occurrence of the value "uncertain," also helps gain insight into physician documentation efficacy within the facility. By utilizing this approach, therefore, the study serves the twin purpose of 1) addressing the gap in the literature with respect to large-scale studies of "coding for quality," and 2) providing insight into the structural characteristics of institutions that are likely facing organizational challenges of physician documentation from the perspective of quality measurement.


Henle fiber layer phase retardation changes associated with age-related macular degeneration.
To quantify and compare phase retardation amplitude and regularity associated with the Henle fiber layer (HFL) between nonexudative AMD patients and age-matched controls using scanning laser polarimetry (SLP) imaging. A scanning laser polarimeter was used to collect 15 × 15° macular-centered images in 25 patients with nonexudative AMD and 25 age-matched controls. Raw image data were used to compute macular phase retardation maps associated with the HFL. Consecutive, annular regions of interest from 0.5 to 3.0° eccentricity, centered on the fovea, were used to generate intensity profiles from phase retardation data and analyzed with two complementary techniques: a normalized second harmonic frequency (2f) of the fast Fourier Transform (FFT) analysis and a curve fitting analysis using a 2f sine function. Paired t-tests were used to compare the normalized 2f FFT magnitude at each eccentricity between the two groups, the eccentricity that yielded the maximum normalized 2f FFT between paired individuals across the two groups, and curve fitting RMS error at each eccentricity between the two groups. Normalized 2f FFT components were lower in the AMD group at each eccentricity, with no difference between the two groups in the maximum normalized 2f FFT component eccentricity. The root-mean-square (RMS) error from curve fitting was significantly higher in the AMD group. Phase retardation changes in the central macula indicate loss and/or structural alterations to central cone photoreceptors in nonexudative AMD patients. Scanning laser polarimetry imaging is a noninvasive method for quantifying cone photoreceptor changes associated with central macular disease.


Phase modifiers promote efficient production of hydroxymethylfurfural from fructose.
Furan derivatives obtained from renewable biomass resources have the potential to serve as substitutes for the petroleum-based building blocks that are currently used in the production of plastics and fine chemicals. We developed a process for the selective dehydration of fructose to 5-hydroxymethylfurfural (HMF) that operates at high fructose concentrations (10 to 50 weight %), achieves high yields (80% HMF selectivity at 90% fructose conversion), and delivers HMF in a separation-friendly solvent. In a two-phase reactor system, fructose is dehydrated in the aqueous phase with the use of an acid catalyst (hydrochloric acid or an acidic ion-exchange resin) with dimethylsulfoxide and/or poly(1-vinyl-2-pyrrolidinone) added to suppress undesired side reactions. The HMF product is continuously extracted into an organic phase (methylisobutylketone) modified with 2-butanol to enhance partitioning from the reactive aqueous solution.


Evaluation of multidimensional COPD-related subjective fatigue following a pulmonary rehabilitation programme.
Subjective fatigue has been recognised as an important, multi-component symptom in COPD. Pulmonary Rehabilitation (PR) improves fatigue component of the Chronic Respiratory Questionnaire, a quality of life (QoL) measure. However, it is not clear if all fatigue dimensions are affected equally. This study aims to evaluate changes in subjective multidimensional fatigue among people with COPD who participated in PR. Thirty seven stable COPD patients were recruited; 23 patients (15 male) mean age 68.5 (range 49-86) yrs, mean (SD) %predicted FEV1 45.3 (19.8); completed 7 weeks of PR. Assessments (pre and post PR) consisted of the Multidimensional Fatigue Inventory (MFI-20), QoL (SGRQ), Anxiety and Depression (HADS), the London Chest Activity of Daily Living Scale (LCADL), muscle strength, incremental (ISWT) and endurance (ESWT) shuttle walk tests. The differences between pre and post PR fatigue were tested using Wilcoxon's test and relationships with other outcomes were examined using Spearman's correlation. There were statistically significant improvements in Reduced Activity (RA) (p = 0.01), General (GF) (p < 0.01) and Physical Fatigue (PF) (p = 0.03) components of MFI-20 after PR, but there were no differences in Motivation or Mental Fatigue (p > 0.05). There were significant improvements in ISWT (p < 0.05), ESWT (p < 0.01) and muscle strength (p = 0.03). Statistically significant correlations (p < 0.05) were found between changes in GF and in both ISWT (r = -0.43) and SGRQ impact (r = 0.46); and between RA and ESWT changes (r = -0.45). Some dimensions of fatigue in COPD are modifiable by a 7-week PR programme. Change in fatigue dimensions in COPD may be associated with a change in maximal or endurance walking distances or QoL.


Conditional TNF-α Overexpression in the Tooth and Alveolar Bone Results in Painful Pulpitis and Osteitis.
Tumor necrosis factor-α (TNF-α) is a proalgesic cytokine that is commonly expressed following tissue injury. TNF-α expression not only promotes inflammation but can also lead to pain hypersensitivity in nociceptors. With the established link between TNF-α and inflammatory pain, we identified its increased expression in the teeth of patients affected with caries and pulpitis. We generated a transgenic mouse model (TNF-α(glo)) that could be used to conditionally overexpress TNF-α. These mice were bred with a dentin matrix protein 1 (DMP1)-Cre line for overexpression of TNF-α in both the tooth pulp and bone to study oral pain that would result from subsequent development of pulpitis and bone loss. The resulting DMP1/TNF-α(glo) mice show inflammation in the tooth pulp that resembles pulpitis while also displaying periodontal bone loss. Inflammatory infiltrates and enlarged blood vessels were observed in the tooth pulp. Pulpitis and osteitis affected the nociceptive neurons innervating the orofacial region by causing increased expression of inflammatory cytokines within the trigeminal ganglia. With this new mouse model morphologically mimicking pulpitis and osteitis, we tested it for signs of oral pain with an oral function assay (dolognawmeter). This assay/device records the time required by a mouse to complete a discrete gnawing task. The duration of gnawing required by the DMP1/TNF-α(glo) mice to complete the task was greater than that for the controls; extended gnaw time in a dolognawmeter indicates reduced orofacial function. With the DMP1/TNF-α(glo) mice, we have shown that TNF-α expression alone can produce inflammation similar to pulpitis and osteitis and that this mouse model can be used to study dental inflammatory pain.


Myeloid-Derived Suppressor Cells.
Myeloid cells developed evolutionarily as a major mechanism to protect the host. They evolved as a critical barrier against infections and are important contributors to tissue remodeling. However, in cancer, myeloid cells are largely converted to serve a new master-tumor cells. This process is epitomized by myeloid-derived suppressor cells (MDSC). These cells are closely related to neutrophils and monocytes. MDSCs are not present in the steady state of healthy individuals and appear in cancer and in pathologic conditions associated with chronic inflammation or stress. These cells have emerged as an important contributor to tumor progression. Ample evidence supports a key role for MDSCs in immune suppression in cancer, as well as their prominent role in tumor angiogenesis, drug resistance, and promotion of tumor metastases. MDSCs have a fascinating biology and are implicated in limiting the effects of cancer immunotherapy. Therefore, targeting these cells may represent an attractive therapeutic opportunity. Cancer Immunol Res; 5(1); 3-8. ©2016 AACR.


Multimodal primary cancer treatment (adjuvant chemotherapy): current results and future prospects.
In the 1970s chemotherapy has been successfully incorporated into curative primary treatment programs for various adult malignancies so that it is no longer solely palliative treatment for advanced disease. For at least three malignancies and tentatively a fourth (breast and colon carcinoma, osteosarcoma, and melanoma), certain groups of patients have had longer disease-free survival produced by the use of chemotherapy after surgical removal of the primary lesion. The potential impact on cancer mortality from these treatment results is obvious. We review here the fundamental laboratory concepts that have led to human trial of multimodal primary therapy regimens. Data from numerous clinical trials are analyzed, with delineation of the problems encountered in the interpretation of their results.


The cross-national epidemiology of social phobia: a preliminary report.
We present a preliminary report on cross-national rates, age at onset, comorbidity, suicide attempts and symptom profiles of social phobia. These data are based on epidemiologic community surveys using similar methods from the United States, Canada, Puerto Rico and Korea. The lifetime rate of social phobia (cases per 100 persons) varies by country, from 2.6 in the United States to 0.5 in Korea. Rates are higher in females than in males in all countries. The age of onset for any phobia is mid-teens to early twenties, and social phobia usually has its first onset before other psychiatric disorders. Only a third or fewer cases of social phobia are uncomplicated by another psychiatric disorder. The presence of social phobia increases the risk of suicide attempts in persons with any other psychiatric disorder. Symptom profiles vary by country. Our data indicate consistent patterns but different cultural expressions of social phobia in the countries studied. The early age of onset of social phobia followed subsequently by another psychiatric disorder raises the possibility that early treatment of social phobia could prevent the onset of other psychiatric disorders.


Carryover Effects of Acute DEHP Exposure on Ovarian Function and Oocyte Developmental Competence in Lactating Cows.
We examined acute exposure of Holstein cows to di(2-ethylhexyl) phthalate (DEHP) and its carryover effects on ovarian function and oocyte developmental competence. Synchronized cows were tube-fed with water or 100 mg/kg DEHP per day for 3 days. Blood, urine and milk samples were collected before, during and after DEHP exposure to examine its clearance pattern. Ovarian follicular dynamics was monitored through an entire estrous cycle by ultrasonographic scanning. Follicular fluids were aspirated from the preovulatory follicles on days 0 and 29 of the experiment and analyzed for phthalate metabolites and estradiol concentration. The aspirated follicular fluid was used as maturation medium for in-vitro embryo production. Findings revealed that DEHP impairs the pattern of follicular development, with a prominent effect on dominant follicles. The diameter and growth rate of the first- and second-wave dominant follicles were lower (P < 0.05) in the DEHP-treated group. Estradiol concentration in the follicular fluid was lower in the DEHP-treated group than in controls, and associated with a higher number of follicular pathologies (follicle diameter >25 mm). The pattern of growth and regression of the corpus luteum differed between groups, with a lower volume in the DEHP-treated group (P < 0.05). The follicular fluid aspirated from the DEHP-treated group, but not the controls, contained 23 nM mono(2-ethylhexyl) phthalate. Culturing of cumulus oocyte complexes in the follicular fluid aspirated from DEHP-treated cows reduced the proportion of oocytes progressing to the MII stage, and the proportions of 2- to 4-cell-stage embryos (P < 0.04) and 7-day blastocysts (P < 0.06). The results describe the risk associated with acute exposure to DEHP and its deleterious carryover effects on ovarian function, nuclear maturation and oocyte developmental competence.


Cationic liposomes loaded with a synthetic long peptide and poly(I:C): a defined adjuvanted vaccine for induction of antigen-specific T cell cytotoxicity.
For effective cancer immunotherapy by vaccination, co-delivery of tumour antigens and adjuvants to dendritic cells and subsequent activation of antigen-specific cytotoxic T cells (CTLs) is crucial. In this study, a synthetic long peptide (SLP) harbouring the model CTL epitope SIINFEKL was encapsulated with the TLR3 ligand poly(inosinic-polycytidylic acid) (poly(I:C)) in cationic liposomes consisting of DOTAP and DOPC. The obtained particles were down-sized to about 140 nm (measured by dynamic light scattering) and had a positive zeta-potential of about 26 mV (according to laser Doppler electrophoresis). SLP loading efficiency was about 40% as determined by HPLC. Poly(I:C) loading efficiency was about 50%, as assessed from the fluorescence intensity of fluorescently labelled poly(I:C). Immunogenicity of the liposomal SLP vaccine was evaluated in vitro by its capacity to activate dendritic cells (DCs) and present the processed SLP to SIINFEKL-specific T cells. The effectiveness of the vaccine to activate CD8(+) T cells was analysed in vivo after intradermal and subcutaneous immunisation in mice, by measuring antigen-specific T cells in blood and spleens and assessing their functionality by cytokine production and in vivo cytotoxicity. The liposomal formulation efficiently delivered the SLP to DCs in vitro and induced a functional CD8(+) T cell immune response in vivo to the CTL epitope present in the SLP. The SLP-specific CD8(+) T cell frequency induced by the poly(I:C)-adjuvanted liposomal SLP formulation showed an at least 25 fold increase over the T cell frequency induced by the poly(I:C)-adjuvanted soluble SLP. In conclusion, cationic liposomes loaded with SLP and poly(I:C) have potential as a powerful therapeutic cancer vaccine formulation.


Dissociation between plasma atrial natriuretic peptide levels and urinary sodium excretion after intravenous saline infusion in normal man.
1. Plasma immunoreactive atrial natriuretic peptide (ANP) and urinary sodium excretion were measured in six normal male subjects before, during and for 195 min after a 60 min infusion of 2 litres of saline (0.9% NaCl, 308 mmol of Na+). 2. During the saline infusion, there was a significant increase in plasma ANP and urinary sodium excretion and a significant decrease in plasma renin activity, aldosterone, albumin, creatinine and packed cell volume. 3. The maximal rise in mean plasma ANP occurred 15 min after stopping the infusion and the maximal rise in mean urinary sodium excretion in the collection period 30 min later. 4. Plasma ANP then decreased so that by the end of the study the level was the same as before the saline infusion. However, at this time, 195 min after the saline infusion was stopped, there was still a net positive sodium balance of 220 mmol and urinary sodium excretion remained significantly elevated. 5. Our results are compatible with the concept that increased ANP secretion may play a role in the immediate increase in sodium excretion after a saline load. However, they also suggest that other mechanisms may be more important for the longer term increase in sodium excretion.


Radio-frequency spectrum analysis of a jittery train after a second-order dispersive Talbot line.
The power spectral density of the intensity of coherent Gaussian pulse trains suffering timing jitter after a dispersive line with arbitrary first- (beta(2)) and second-order (beta(3)) dispersion is computed in the small-signal approximation. Due to timing jitter noise, the initial radio-frequency spectrum shows noise bands whose bandwidth and position depend, respectively, on the jitter's standard deviation and on the jitter's pulse-to-pulse correlation. After setting the accumulated first-order dispersion to Talbot conditions, it is shown that the influence on the noise spectrum is a multiplicative factor with a multiple-bandpass structure. This factor depends on both the dispersive characteristics of the line and the pulse parameters, but not on the timing jitter's correlation properties, and represents the filtering mechanism responsible for Talbot repetition-rate multiplication. It is shown that the integer or fractional temporal Talbot effect does not worsen the timing properties of the initial train. In addition, and depending on the type of jitter correlation, the pulse width, and the total dispersion, it is shown that the temporal Talbot effect may lead to significant jitter reduction. The theory is exemplified by use of simulations. The applicability of the model to practical situations is also analyzed.


Large-bandwidth, low-loss, efficient mode mixing using long-period mechanical gratings.
We propose a new architecture for using long-period fiber gratings (LPGs) to induce strong mode mixing with low loss for space-division multiplexing. In this architecture, LPGs are installed in step-index (SI) few-mode fibers that support more modes than the transmission fiber. Such a design could significantly reduce losses due to coupling from the highest-order mode group to cladding modes. In our experiment, efficient mixing of three spatial modes over a broad bandwidth was achieved by a mechanical long-period grating on a SI fiber that supports eight spatial modes. The insertion loss, including two splice losses, is less than 0.5 dB, and the coupling matrix and mode-dependent loss (MDL) are characterized experimentally for the first time, to the best of our knowledge. Strong mixing between LP01 and LP11 for a whole C band is demonstrated, and MDL introduced to the system is negligible.


Correlates of overweight and obesity in 644 patients with bipolar disorder.
Overweight and obesity are common clinical problems encountered in the treatment of bipolar disorder. We therefore assessed the prevalence and clinical correlates of overweight, obesity, and extreme obesity in 644 bipolar patients. 644 outpatients with DSM-IV bipolar disorder in the Stanley Foundation Bipolar Treatment Outcomes Network were evaluated with structured diagnostic interviews and clinician- and self-administered questionnaires to determine bipolar disorder diagnoses, demographic and historical illness characteristics, comorbid Axis I diagnoses, medical histories, health habits, and body mass indices (BMMs). Fifty-eight percent of the patients with bipolar disorder were overweight, 21% were obese, and 5% were extremely obese. American patients had significantly higher mean (p < .0001) BMIs and significantly higher rates of obesity (p < .001) and extreme obesity (p < .001) than European patients. Significant associations (p < or = .001) were found between overweight, obesity. and extreme obesity and gender, age, income level, comorbid binge-eating disorder, hypertension, arthritis, diabetes mellitus, exercise habits, and coffee consumption. Current BMI and weight were each correlated with the number of weight gain-associated psychotropics to which patients had been exposed. Multinomial logistic regression (adjusted for site and eating disorder diagnosis and corrected for multiple comparisons) showed that (1) overweight was significantly associated with male gender and hypertension (p < .001), (2) obesity was significantly associated with hypertension (p < .001), and (3) extreme obesity was significantly associated with hypertension and arthritis (p < .001). Overweight, obesity, and extreme obesity were common in this group of bipolar patients, although it was unclear that their prevalence rates were truly elevated, because overweight and obesity are increasingly common public health problems among the general population. Correlates of overweight and obesity in bipolar disorder include patient and treatment variables such as gender, geographical location, comorbid binge-eating disorder, age, income level, degree of exposure to weight gain-associated psychotropics, medical disorders associated with obesity, and health habits.


High resolution flow cytometric analysis of electronic nuclear volume and DNA content in normal and abnormal human tissue.
The NPE Analyzer flow cytometer can simultaneously analyze the electronic nuclear volume (ENV) and DNA content of cells. This study describes the schematics, resolution, reproducibility, and sensitivity of biological standards analyzed on this unit. Calibrated beads and biological standards (lymphocytes, trout erythrocytes [TRBC], calf thymocytes, and tumor cells) were analyzed for ENV versus DNA content. Parallel data (forward scatter versus DNA) from a conventional flow cytometer were obtained. ENV linearity studies yielded an R value of 0.999. TRBC had a coefficient of variation (CV) of 1.18 +/- 0.13. DNA indexes as low as 1.02 were detectable. DNA content of lymphocytes from 42 females was 1.9% greater than that for 60 males, with a noninstrumental variability in total DNA content of 0.5%. The ENV/DNA ratio was constant in 15 normal human tissue samples, but differed in the four animal species tested. The ENV/DNA ratio for a hypodiploid breast carcinoma was 2.3 times greater than that for normal breast tissue. The high-resolution ENV versus DNA analyses are highly reliable, sensitive, and can be used for the detection of near-diploid tumor cells that are difficult to identify with conventional cytometers. ENV/DNA ratio may be a useful parameter for detection of aneuploid populations.


Eosinophilic dysplasia of the gallbladder: a hitherto undescribed variant identified in association with a "porcelain" gallbladder.
Non-mass forming, neoplastic intraepithelial proliferations (dysplasia) represent the most well-accepted precursor lesions to gallbladder adenocarcinomas. They are typically small, localized, grossly unrecognizable lesions that have been identified in the epithelium adjacent to up to 79% of gallbladder adenocarcinomas. Morphologic variants that have been reported include flat, micropapillary, papillary and cribriform. We have recently encountered a morphologically distinctive, previously unreported lesion to which we have applied the designation eosinophilic dysplasia. This lesion was identified in a gallbladder with diffuse mural fibrosis and calcification (porcelain gallbladder). The dysplastic focus was confined to one tissue section, and was comprised of a localized true papilla [i.e with a fibrovascular core], measuring approximately 1.2 mm in greatest dimension and an adjacent, flat, 7-cell epithelial segment. These foci were lined by cells displaying significant nuclear enlargement [1.5-4 times the adjacent benign cells], nuclear pleomorphism, occasional multinucleation, hyperchromasia and nuclear membrane irregularities. Nucleoli were present but inconspicuous. These cells also showed voluminous eosinophilic to granular cytoplasm, such that the overall nuclear-to-cytoplasmic ratio was generally not increased. The cells displayed diffuse and marked nuclear immunoreactivity for p53, and approximately 70% of the cells showed nuclear positivity for Ki-67. The cells were also positive for cytokeratin 7 and were entirely negative for carcinoembryonic antigen (CEA) and chromogranin A. The cells of the adjacent normal epithelium were positive for cytokeratin 7 and CEA, negative for p53 and chromogranin A and showed a Ki-67 labeling index of <10%. Marked overexpression of the p53 protein as well as its high proliferative index are strong arguments in favor of the dysplastic nature of this lesion. However, further studies are required to elucidate its true clinical significance and to determine whether or not its association with a porcelain gallbladder, as noted herein, is entirely fortuitous. However, such studies can only be performed with an increased recognition by practitioners of this distinctive variant.


Publication bias in clinical research.
In a retrospective survey, 487 research projects approved by the Central Oxford Research Ethics Committee between 1984 and 1987, were studied for evidence of publication bias. As of May, 1990, 285 of the studies had been analysed by the investigators, and 52% of these had been published. Studies with statistically significant results were more likely to be published than those finding no difference between the study groups (adjusted odds ratio [OR] 2.32; 95% confidence interval [Cl] 1.25-4.28). Studies with significant results were also more likely to lead to a greater number of publications and presentations and to be published in journals with a high citation impact factor. An increased likelihood of publication was also associated with a high rating by the investigator of the importance of the study results, and with increasing sample size. The tendency towards publication bias was greater with observational and laboratory-based experimental studies (OR = 3.79; 95% Cl = 1.47-9.76) than with randomised clinical trials (OR = 0.84; 95% Cl = 0.34-2.09). We have confirmed the presence of publication bias in a cohort of clinical research studies. These findings suggest that conclusions based only on a review of published data should be interpreted cautiously, especially for observational studies. Improved strategies are needed to identify the results of unpublished as well as published studies.


Signal transduction pathways that function in both development and innate immunity.
C. elegans is developing in importance as a model for innate immunity. Several signaling pathways are known to be required for immune responses to a diverse range of pathogens, including the insulin signaling, p38 MAP kinase and transforming growth factor-beta pathways. These pathways also have roles during development, which can complicate the analysis of their functions in immunity. Recent studies have suggested that immunity in C. elegans is integrated across the organism by both paracrine and neuronal communication, showing the complexity of the immune system in this organism.


Targeting vascular pericytes in hypoxic tumors increases lung metastasis via angiopoietin-2.
Strategies to target angiogenesis include inhibition of the vessel-stabilizing properties of vascular pericytes. Pericyte depletion in early-stage non-hypoxic tumors suppressed nascent angiogenesis, tumor growth, and lung metastasis. In contrast, pericyte depletion in advanced-stage hypoxic tumors with pre-established vasculature resulted in enhanced intra-tumoral hypoxia, decreased tumor growth, and increased lung metastasis. Furthermore, depletion of pericytes in post-natal retinal blood vessels resulted in abnormal and leaky vasculature. Tumor transcriptome profiling and biological validation revealed that angiopoietin signaling is a key regulatory pathway associated with pericyte targeting. Indeed, pericyte targeting in established mouse tumors increased angiopoietin-2 (ANG2/Angpt2) expression. Depletion of pericytes, coupled with targeting of ANG2 signaling, restored vascular stability in multiple model systems and decreased tumor growth and metastasis. Importantly, ANGPT2 expression correlated with poor outcome in patients with breast cancer. These results emphasize the potential utility of therapeutic regimens that target pericytes and ANG2 signaling in metastatic breast cancer.


FMR1 RNA within the intranuclear inclusions of fragile X-associated tremor/ataxia syndrome (FXTAS).
Fragile X-associated tremor/ataxia syndrome (FXTAS) is a recently identified neurodegenerative disorder affecting older adult males with premutation alleles of the fragile X mental retardation 1 (FMR1) gene. The principal clinical features of FXTAS include progressive intention tremor, gait ataxia, parkinsonism, and autonomic dysfunction. The disorder affects at least one-third of carrier males over 50 years of age and, with an estimated carrier frequency of approximately 1/800 males, is likely to be one of the most common heritable forms of tremor and ataxia among older adult males in the general population. Brains from all FXTAS cases examined to date (10/10) possess numerous ubiquitin-positive intranuclear inclusions in broad distribution throughout the cerebrum and brainstem. The absence of either the neurodegenerative disorder or inclusions among adults with fragile X syndrome (who lack the FMR1 protein), coupled with elevated FMR1 mRNA with expanded CGG repeats in premutation carriers, has led us to propose an RNA toxic gain-of-function model for FXTAS. Consistent with this model, we have now identified FMR1 mRNA within the intranuclear inclusions isolated from post-mortem (FXTAS) brain tissue.


Using computer modeling to help identify patient subgroups in clinical data repositories.
The ability to accurately and efficiently identify patient cases of interest in a hospital information system has many important clinical, research, educational and administrative uses. The identification of cases of interest sometimes can be difficult. This paper describes a two-stage method for searching for cases of interest. First, a Boolean search is performed using coded database variables. The user classifies the retrieved cases as being of interest or not. Second, based on the user-classified cases, a computer model of the patient cases of interest is constructed. The model is then used to help locate additional cases. These cases provide an augmented training set for constructing a new computer model of the cases of interest. This cycle of modeling and user classification continues until halted by the user. This paper describes a pilot study in which this method is used to identify the records of patients who have venous thrombosis. The results indicate that computer modeling enhances the identification of patient cases of interest.


Management of pediatric patients with irreducible atlantoaxial dislocation: transoral anterior release, reduction, and fixation.
Although transoral atlantoaxial reduction plate (TARP) surgery has been confirmed to be safe and effective for adults who have irreducible atlantoaxial dislocation (IAAD) with or without basilar invagination or upper cervical revision surgery, it is rarely used to treat these disorders in children. The authors of this study aimed to report on the use of the anterior technique in treating pediatric IAAD. In this retrospective study, the authors identified 8 consecutive patients with IAAD who had undergone surgical reduction at a single institution in the period between January 2011 and June 2104. The patients consisted of 5 males and 3 females. Three had os odontoideum, 2 had basilar invagination, and the other 3 experienced atlantoaxial rotatory fixed dislocation (AARFD). They were all treated using transoral anterior release, reduction, and fusion with the TARP. Preoperative and postoperative CT scans and MR images were obtained. American Spinal Injury Association (ASIA) Impairment Scale grades were determined. All symptoms were relieved in all 8 patients but to varying degrees. Intraoperative loose reduction and fixation of C1-2 were achieved in one stage. The 4 patients with preoperative neurological deficits were significantly improved after surgery, and their latest follow-ups indicated that their ASIA Impairment Scale grades had improved to E. Postoperative pneumonia occurred in 1 patient but was under complete control after anti-infective therapy and fiber optic-guided sputum suction. One-stage transoral anterior release, reduction, and fixation is an effective, reliable, and safe means of treating pediatric IAAD. The midterm clinical results are satisfactory, with the technique eliminating the need for interval traction and/or second-stage posterior instrumentation and fusion.


Effect of CYP2D6*10 and APOE polymorphisms on the efficacy of donepezil in patients with Alzheimer's disease.
The aim of this study was to evaluate the effect of CYP2D6*10 and APOE polymorphisms on both steady-state plasma concentrations (Cp) and clinical response of donepezil in patients with mild-to-moderate Alzheimer's disease (AD). A total of 110 Chinese AD patients participated in this study. Patients were treated with 5 to 10 mg of donepezil daily for 6 months. The genotypes of CYP2D6*10 and APOE were analyzed by polymerase chain reaction-restriction fragment length polymorphism. The steady-state Cp of donepezil was measured by high-performance liquid chromatography-tandem mass spectrometric assay method. The cognition of patients was evaluated at baseline and at 6-month follow-up by Mini-Mental Status Examination and Alzheimer Disease Assessment Scale-Cognitive subscale. At 6-month follow-up, 56 of 96 patients (58.3%) were evaluated as responders and 40 patients (41.7%) as nonresponders to donepezil treatment. A significantly higher frequency of patients with genotypes CYP2D6*1/*10 and *10/*10 were found in responders than in nonresponders (P < 0.05). Besides, patients with CYP2D6*1/*10 and *10/*10 genotypes had higher Cp of donepezil and improved cognition scores than those with CYP2D6*1/*1 genotype (P < 0.05). However, the frequency of APOE [Latin Small Letter Open E]4 carriers and noncarriers showed no difference between the 2 groups (P > 0.05). AD patients with mutant allele (*10) in CYP2D6 gene may respond better to donepezil than those with wild allele (*1). We did not find the relationship between APOE [Latin Small Letter Open E]4 status and the efficacy of donepezil in our study.


Spatial symmetries in vestibular projections to the uvula-nodulus.
The discharge of secondary vestibular neurons relays the activity of the vestibular endorgans, occasioned by movements in three-dimensional physical space. At a slightly higher level of analysis, the discharge of each secondary vestibular neuron participates in a multifiber projection or pathway from primary afferents via the secondary neurons to another neuronal population. The logical organization of this projection determines whether characteristics of physical space are retained or lost. The logical structure of physical space is standardly expressed in terms of the mathematics of group theory. The logical organization of a projection can be compared to that of physical space by evaluating its symmetry group. The direct projection from the semicircular canal nerves via the vestibular nuclei to neck motor neurons has a full three-dimensional symmetry group, allowing it to maintain a three-dimensional coordinate frame. However, a projection may embed only a subgroup of the symmetry group of physical space, which incompletely mirrors the properties of physical space. The major visual and vestibular projections in the rabbit via the inferior olive to the uvula-nodulus carry three degrees of freedom-rotations about one vertical and two horizontal axes-but do not have full three dimensional symmetry. Instead, the vestibulo-olivo-nodular projection has symmetries corresponding to a product of two-dimensional vestibular and one-dimensional optokinetic spaces. This combination of projection symmetries provides the foundation for distinguishing horizontal from vertical rotations within a three dimensional space. In this study, we evaluate the symmetry group given by the physiological organization of the vestibulo-olivo-nodular projection. Although it acts on the same sets of elements and mirrors the rotations that occur in physical space, the physiological transformation group is distinct from the spatial group. We identify symmetries as products of physiological and spatial transformations. The symmetry group shapes the information the projection conveys to the uvula-nodulus; this shaping may depend on a physiological choice of generators, in the same way that function depends on the physiological choice of coordinates. We discuss the implications of the symmetry group for uvula-nodulus function, evolution, and functions of the vestibular system in general.


Cutting edge: V alpha 14-J alpha 281 NKT cells naturally regulate experimental autoimmune encephalomyelitis in nonobese diabetic mice.
Although deficiencies in the NKT cell population have been observed in multiple sclerosis and mouse strains susceptible to experimental autoimmune encephalomyelitis (EAE), little is known about the function of these cells in CNS autoimmunity. In this work we report that TCR Valpha14-Jalpha281 transgenic nonobese diabetic mice, which are enriched in CD1d-restricted NKT cells, are protected from EAE. The protection is associated with a striking inhibition of Ag-specific IFN-gamma production in the spleen, implying modulation of the encephalitogenic Th1 response. This modulation is independent of IL-4 because IL-4-deficient Valpha14-Jalpha281 mice are still protected against EAE and independent of NKT cell-driven Th1 to Th2 deviation, because no increased autoantigen-specific Th2 response was observed in immunized Valpha14-Jalpha281 transgenic mice. Our findings indicate that enrichment and/or stimulation of CD1d-dependent NKT cells may be used as a novel strategy to treat CNS autoimmunity.


A set of novel microsatellite markers developed for an economically important tree, Dracontomelon duperreanum, in China.
Dracontomelon duperreanum, the most representative species of the family Anacardiaceae, is an important multipurpose tree in China and Vietnam. However, no genetic diversity studies have been reported on this species. In this study, we identified 11 microsatellite markers for D. duperreanum by using the restriction-site-associated DNA sequencing (RAD-seq) method and examined their polymorphisms in 22 samples obtained from the South China Botanical Garden, South China. We could detect only two or three alleles for each microsatellite marker. The mean observed and expected heterozygosities were 0.41 and 0.39, respectively, which were lower than those reported for the species with similar life history forms. These relatively low genetic diversities in this common plant species are unexpected and might have resulted from its extensive cultivation. To our knowledge, this is the first report of microsatellite markers in the genus Dracontomelon. These microsatellite markers will be valuable for studying the genetic diversities and structures in D. duperreanum and other Dracontomelon species.


Demethoxycurcumin: A naturally occurring curcumin analogue for treating non-cancerous diseases.
Turmeric extracts contain three primary compounds, which are commonly referred to as curcuminoids. They are curcumin, demethoxycurcumin (DMC), and bisdemethoxycurcumin. While curcumin has been the most extensively studied of the curcuminoids, it suffers from low overall oral bioavailability due to extremely low absorption as a result of low water solubility and instability at acidic pH, as well as rapid metabolism and clearance from the body. However, DMC, which lacks the methoxy group on the benzene ring of the parent structure, has much greater chemical stability at physiological pH and has been recently reported to exhibit antitumor properties. However, the treatment of noncancerous diseases with DMC has not been comprehensively reviewed. Therefore, here we evaluate published scientific literature on the therapeutic properties of DMC. The beneficial pharmacological actions of DMC include anti-inflammatory, neuroprotective, antihypertensive, antimalarial, antimicrobial, antifungal, and vasodilatory properties. In addition, DMC's ability to ameliorate the effects of free radicals and an environment characterized by oxidative stress caused by the accumulation of advanced glycation end-products associated with diabetic nephropathy, as well as DMC's capacity to inhibit the migration and proliferation of vascular smooth muscle cells following balloon angioplasty are also addressed. This review collates the available literature regarding the therapeutic possibilities of DMC in noncancerous conditions.


Serum peptide profiles during progression of chronic hepatitis B virus infection to liver failure.
Chronic hepatitis B virus (HBV)-infected patients with liver failure have a poor prognosis, and no satisfactory biomarkers are available for diagnosis before the end-stage. We explored serum peptide profiling for diagnosis and prediction of progression to liver failure in HBV-infected patients. Serum samples (164) from healthy subjects (n = 20), or subjects with chronic hepatitis B without cirrhosis and liver failure [chronic hepatitis B subjects without cirrhosis and liver failure (CHB); n = 33], with compensated liver cirrhosis (compensated liver cirrhosis (LC); n = 35), with acute-on-chronic liver failure [acute-on-chronic liver failure (ACLF); n = 38] or with chronic liver failure [chronic liver failure (CLF), n = 38] were applied to ClinProt magnetic beads, and bound peptides/proteins were analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Our classification diagnostic models of liver disease were generated based on the Genetic Algorithm (GA) and Quick Classifier Algorithm (QC). Differentially expressed peptides were found among all test groups, with patterns of difference that readily distinguished between healthy and various HBV-associated liver disease samples. The model generated seven characteristic peptide peaks at 4053 m/z, 3506 m/z, 4963 m/z, 9289 m/z, 2628 m/z, 3193 m/z and 6432 m/z, giving overall predictive capability of 54.27%. Two-way comparisons of LC, ACLF or CLF vs CHB had predictive capabilities of 79.8%, 91.41% and 97.99%, respectively. Comparisons of ACLF or CLF vs LC were predictive at 87.72% and 82.18%, respectively and ACLF vs CLF was predictive at 75.05%. These classification diagnostic models generated by different peptide peaks were further validated in blinded tests with 67-100% accuracy. Serum peptide patterns vary during progression of chronic HBV infection to liver failure and may be used to distinguish different stages of the disease.


Estimation of Regional Economic Development Indicator from Transportation Network Analytics.
With the booming economy in China, many researches have pointed out that the improvement of regional transportation infrastructure among other factors had an important effect on economic growth. Utilizing a large-scale dataset which includes 3.5 billion entry and exit records of vehicles along highways generated from toll collection systems, we attempt to establish the relevance of mid-distance land transport patterns to regional economic status through transportation network analyses. We apply standard measurements of complex networks to analyze the highway transportation networks. A set of traffic flow features are computed and correlated to the regional economic development indicator. The multi-linear regression models explain about 89% to 96% of the variation of cities' GDP across three provinces in China. We then fit gravity models using annual traffic volumes of cars, buses, and freight trucks between pairs of cities for each province separately as well as for the whole dataset. We find the temporal changes of distance-decay effects on spatial interactions between cities in transportation networks, which link to the economic development patterns of each province. We conclude that transportation big data reveal the status of regional economic development and contain valuable information of human mobility, production linkages, and logistics for regional management and planning. Our research offers insights into the investigation of regional economic development status using highway transportation big data.


Memory and complex dynamics in cardiac Purkinje fibers.
The contribution of cumulative changes in action potential duration (memory) to complex cellular electrophysiological behavior was investigated in canine cardiac Purkinje fibers. Complex behavior induced during constant pacing was caused by reciprocal interactions between the time to full repolarization (TFR), where TFR = response duration + latency, and the diastolic interval (DI). The relationship between TFR and the preceding DI during complex behavior differed from that obtained using a standard restitution protocol. In particular, higher-order periodicities and chaos were produced in fibers in which the restitution curve lacked the prerequisites for such behavior. To investigate whether shifts in the restitution curve might be expected during rapid pacing, the relationship between TFR of a test response (TFR(n + 1)) and the immediately preceding response (TFR(n)) was determined. For any fixed DI(n), reduction of TFR(n) from 240 to 130 ms was accompanied by a corresponding reduction of TFR(n + 1), whereas as TFR(n) was reduced further to 120 ms, TFR(n + 1) increased. Because of the dependence of TFR(n + 1) on TFR(n) (memory) and on the preceding DI(n) (restitution), the slope of the low-dimensional relationship between TFR(n + 1) and DI(n) at a constant pacing cycle length depended on the slopes of the restitution and memory functions. These results suggest that rapid accumulation and dissipation of memory may contribute importantly to complex electrical behavior in cardiac tissue.


Social isolation from childhood to mid-adulthood: is there an association with older brain age?
Older brain age - as estimated from structural MRI data - is known to be associated with detrimental mental and physical health outcomes in older adults. Social isolation, which has similar detrimental effects on health, may be associated with accelerated brain aging though little is known about how different trajectories of social isolation across the life course moderate this association. We examined the associations between social isolation trajectories from age 5 to age 38 and brain age assessed at age 45. We previously created a typology of social isolation based on onset during the life course and persistence into adulthood, using group-based trajectory analysis of longitudinal data from a New Zealand birth cohort. The typology comprises four groups: 'never-isolated', 'adult-only', 'child-only', and persistent 'child-adult' isolation. A brain age gap estimate (brainAGE) - the difference between predicted age from structural MRI date and chronological age - was derived at age 45. We undertook analyses of brainAGE with trajectory group as the predictor, adjusting for sex, family socio-economic status, and a range of familial and child-behavioral factors. Older brain age in mid-adulthood was associated with trajectories of social isolation after adjustment for family and child confounders, particularly for the 'adult-only' group compared to the 'never-isolated' group. Although our findings are associational, they indicate that preventing social isolation, particularly in mid-adulthood, may help to avert accelerated brain aging associated with negative health outcomes later in life.


A Confined Fabrication of Perovskite Quantum Dots in Oriented MOF Thin Film.
Organic-inorganic hybrid lead organohalide perovskites are inexpensive materials for high-efficiency photovoltaic solar cells, optical properties, and superior electrical conductivity. However, the fabrication of their quantum dots (QDs) with uniform ultrasmall particles is still a challenge. Here we use oriented microporous metal-organic framework (MOF) thin film prepared by liquid phase epitaxy approach as a template for CH3NH3PbI2X (X = Cl, Br, and I) perovskite QDs fabrication. By introducing the PbI2 and CH3NH3X (MAX) precursors into MOF HKUST-1 (Cu3(BTC)2, BTC = 1,3,5-benzene tricarboxylate) thin film in a stepwise approach, the resulting perovskite MAPbI2X (X = Cl, Br, and I) QDs with uniform diameters of 1.5-2 nm match the pore size of HKUST-1. Furthermore, the photoluminescent properties and stability in the moist air of the perovskite QDs loaded HKUST-1 thin film were studied. This confined fabrication strategy demonstrates that the perovskite QDs loaded MOF thin film will be insensitive to air exposure and offers a novel means of confining the uniform size of the similar perovskite QDs according to the oriented porous MOF materials.


Naked five-coordinate Fe(III)(NO) porphyrin complexes: vibrational and reactivity features.
Model ferric heme nitrosyl complexes, [Fe(TPP)(NO)](+) and [Fe(TPFPP)(NO)](+), where TPP is the dianion of 5,10,15,20-tetrakis-phenyl-porphyrin and TPFPP is the dianion of 5,10,15,20-tetrakis-pentafluorophenyl-porphyrin, have been obtained as isolated species by the gas phase reaction of NO with [Fe(III)(TPP)](+) and [Fe(III) (TPFPP)](+) ions delivered in the gas phase by electrospray ionization, respectively. The so-formed nitrosyl complexes have been characterized by vibrational spectroscopy also exploiting (15)N-isotope substitution in the NO ligand. The characteristic NO stretching frequency is observed at 1825 and 1859 cm(-1) for [Fe(III)(TPP)(NO)](+) and [Fe(III)(TPFPP)(NO)](+) ions, respectively, providing reference values for genuine five-coordinate Fe(III)(NO) porphyrin complexes differing only for the presence of either phenyl or pentafluorophenyl substituents on the meso positions of the porphyrin ligand. The vibrational assignment is aided by hybrid density functional theory (DFT) calculations of geometry and electronic structure and frequency analysis which clearly support a singlet spin electronic state for both [Fe(TPP)(NO)](+) and [Fe(TPFPP)(NO)](+) complexes. Both TD-DFT and CASSCF calculations suggest that the singlet ground state is best described as Fe(II)(NO(+)) and that the open-shell AFC bonding scheme contribute for a high-energy excited state. The kinetics of the NO addition reaction in the gas phase are faster for [Fe(III)(TPFPP)](+) ions by a relatively small factor, though highly reliable because of a direct comparative evaluation. The study was aimed at gaining vibrational and reactivity data on five-coordinate Fe(III)(NO) porphyrin complexes, typically transient species in solution, ultimately to provide insights into the nature of the Fe(NO) interaction in heme proteins.


Predicting mesh density for adaptive modelling of the global atmosphere.
The shallow water equations are solved using a mesh of polygons on the sphere, which adapts infrequently to the predicted future solution. Infrequent mesh adaptation reduces the cost of adaptation and load-balancing and will thus allow for more accurate mapping on adaptation. We simulate the growth of a barotropically unstable jet adapting the mesh every 12 h. Using an adaptation criterion based largely on the gradient of the vorticity leads to a mesh with around 20 per cent of the cells of a uniform mesh that gives equivalent results. This is a similar proportion to previous studies of the same test case with mesh adaptation every 1-20 min. The prediction of the mesh density involves solving the shallow water equations on a coarse mesh in advance of the locally refined mesh in order to estimate where features requiring higher resolution will grow, decay or move to. The adaptation criterion consists of two parts: that resolved on the coarse mesh, and that which is not resolved and so is passively advected on the coarse mesh. This combination leads to a balance between resolving features controlled by the large-scale dynamics and maintaining fine-scale features.


Resource needs for addressing noncommunicable disease in low- and middle-income countries: current and future developments.
Low and middle income countries are faced with a range of challenges related to providing efficient and affordable health care. With non-communicable diseases (NCD) on the rise, there is a growing need to be able to estimate resource requirements, costs and expected impact associated with various investment strategies related to prevention and control of NCD. In this article, recently developed costing and health impact models for non-communicable disease are reviewed, with a view to drawing out their main findings as well as methodological limitations. A key shortcoming is that earlier modelling efforts have taken a vertical approach to costing, when in reality a more integrated, horizontal approach is needed in order to effectively plan for scaled-up investment and system development. We subsequently describe how the integration of an NCD module into the joint United Nations OneHealth tool will enable low- and middle-income countries to bring NCD into an integrated process for national strategic health planning.


Caring for dying patients outside special palliative care settings: experiences from a nursing perspective.
The aim of the study was to describe nurses' experiences in caring for gravely ill and dying patients outside special palliative care settings. Tape-recorded qualitative interviews were conducted with a total of nine nurses in primary home care, community care and hospitals. The interviews were analysed according to phenomenological methodology, which resulted in the three common structures: ambition and dedication, everyday encounters, and satisfaction/dissatisfaction. In the 'everyday encounters' structure, the following key constituents emerged: responsibility, cooperation, experience and knowledge, feelings, and time and resources. The results describe the nurses' high ambitions to give dying patients and their relatives high-quality care. Despite this, they experienced greater or lesser degrees of dissatisfaction because of insufficient cooperation, support, time and resources. They experienced satisfaction through contact with patients and relatives, functioning collegial cooperation, and the knowledge, experience and personal growth the care had given them. The results indicate that nurses need the resources such as time, improved methods of communication and cooperation as well as more support in order to give quality palliative care and achieve satisfaction with the outcome. The need for discussion about the conditions for giving palliative care outside the hospices and other special palliative care settings is also elucidated.


Accelerated growth rates of recurrent hepatocellular carcinoma after liver transplantation.
The growth rates of recurrent hepatocellular carcinoma (HCC) after orthotopic liver transplantation (OLTX) were estimated by calculating the tumor doubling time (TDT) in 20 patients. The mean TDT, calculated by multiple measurement of tumor size, was 44.3 +/- 11.3 days (mean +/- standard error) in 12 patients with pulmonary metastasis (range, 10 to 161 days) and 37.6 +/- 8.9 days (range, 7 to 65 days) in 5 patients with liver allograft recurrence. The TDT as estimated by serum alpha-fetoprotein (AFP) levels in 6 patients was 37.3 +/- 10.0 days (range, 12 to 84 days). The mean TDT obtained from 5 control subjects with HCC who were treated with liver resection (without immunosuppression) was 273.8 +/- 79.1 days (range, 82 to 560 days). The disease-free period and survival time after OLTX both correlated well with the TDT (r = 0.546 and r = 0.701, respectively). The patients with fibrolamellar HCC had a greater TDT and a longer survival time than those with nonfibrolamellar HCC. Despite a wide range of TDT in patients who received transplants, their recurrent HCC tumors grew significantly faster than those of patients with the same disease who did not receive transplants. The factors involved in this accelerated growth rate may include the use of immunosuppressive drugs and the consequent suppression of host immunity against the growth of micrometastasis.


Preliminary study of osmotic membrane bioreactor: effects of draw solution on water flux and air scouring on fouling.
Preliminary study on a novel osmotic membrane bioreactor (OMBR) was explored. Objective of this study was to investigate the effects of draw solution on membrane flux and air scouring at the feed side on fouling tendency in a pilot OMBR system composing the anoxic/aerobic and forward osmosis (FO) processes. Domestic sewage was the raw feed, FO membrane from HTI and NaCl/MgSO4 draw solutions were used in the experiments. Fluxes of 3 l/m2/h (LMH) and 7.2 LMH were achieved at osmotic pressure of 5 and 22.4 atm, respectively. No significant flux decline was observed at 3 LMH over 190 h and at 7.2 LMH over 150 h when air scouring was provided at the feed side of the membrane. However, without air scouring, the flux at 22.4 atm osmotic pressure declined by 30% after 195 h and then levelled off. The potential advantages of the fouling reversibility with air scouring under the operating conditions of the pilot OMBR and better water quality in OMBR over the conventional MBR were preliminarily demonstrated.


Supernumerary centrosomes nucleate extra cilia and compromise primary cilium signaling.
The primary cilium is a nexus of cell signaling, and ciliary dysfunction is associated with polycystic kidney disease, retinal degeneration, polydactyly, neural tube defects, and obesity (ciliopathies). Signaling molecules for cilium-associated pathways are concentrated in the cilium, and this is essential for efficient signaling. Cilia are nucleated from centrioles, and aberrant centriole numbers are seen in many cancers and in some ciliopathies. We tested the effect of supernumerary centrioles on cilium function and found that cells with extra centrioles often formed more than one cilium, had reduced ciliary concentration of Smoothened in response to Sonic hedgehog stimulation, and reduced Shh pathway transcriptional activation. This ciliary dilution phenotype was also observed with the serotonin receptor Htr6, fibrocystin PKHD1, and Arl13b. The presence of extra centrioles and cilia disrupted epithelial organization in 3D spheroid culture. Cells mutant for the tuberous sclerosis gene Tsc2 also had extra cilia and diluted ciliary protein. In most cells, extra cilia were clustered and shared the same ciliary pocket, suggesting that the ciliary pocket is the rate-limiting structure for trafficking of ciliary proteins. Thus, extra centrioles and cilia disrupt signaling and may contribute to disease phenotypes.


Extrafoveal attentional capture by object semantics.
There is ongoing debate on whether object meaning can be processed outside foveal vision, making semantics available for attentional guidance. Much of the debate has centred on whether objects that do not fit within an overall scene draw attention, in complex displays that are often difficult to control. Here, we revisited the question by reanalysing data from three experiments that used displays consisting of standalone objects from a carefully controlled stimulus set. Observers searched for a target object, as per auditory instruction. On the critical trials, the displays contained no target but objects that were semantically related to the target, visually related, or unrelated. Analyses using (generalized) linear mixed-effects models showed that, although visually related objects attracted most attention, semantically related objects were also fixated earlier in time than unrelated objects. Moreover, semantic matches affected the very first saccade in the display. The amplitudes of saccades that first entered semantically related objects were larger than 5° on average, confirming that object semantics is available outside foveal vision. Finally, there was no semantic capture of attention for the same objects when observers did not actively look for the target, confirming that it was not stimulus-driven. We discuss the implications for existing models of visual cognition.


Deep sequencing of 10,000 human genomes.
We report on the sequencing of 10,545 human genomes at 30×-40× coverage with an emphasis on quality metrics and novel variant and sequence discovery. We find that 84% of an individual human genome can be sequenced confidently. This high-confidence region includes 91.5% of exon sequence and 95.2% of known pathogenic variant positions. We present the distribution of over 150 million single-nucleotide variants in the coding and noncoding genome. Each newly sequenced genome contributes an average of 8,579 novel variants. In addition, each genome carries on average 0.7 Mb of sequence that is not found in the main build of the hg38 reference genome. The density of this catalog of variation allowed us to construct high-resolution profiles that define genomic sites that are highly intolerant of genetic variation. These results indicate that the data generated by deep genome sequencing is of the quality necessary for clinical use.


Ascorbic acid, metal ions and the superoxide radical.
1. No evidence could be found for production of the superoxide radical, O2-, during autoxidation of ascorbic acid at alkaline pH values. Indeed, ascorbate may be important in protection against O2- genat-d in vivo. 2. Oxidation of ascorbate at pH 10.2 was stimulated by metal ions. Stimulation by Fe2+ was abolished by superoxide dismutase, probably because of generation of O2-- during reduction of O2 by Fe2+, followed by reaction of O2-- with ascorbate. EDTA changed the mechanism of Fe2+-stimulated ascorbate oxidation. 3. Stimulation of ascorbate oxidation by Cu2+ was also decreased by superoxide dismutase, but this appears to be an artifact, since apoenzyme or bovine serum albumin showed similar effects.


Dynamic tomotherapy delivery.
Several dynamic techniques are introduced to speed up TomoTherapy delivery and improve longitudinal conformity. These techniques include dynamic jaws, dynamic couch, and their combinations. In general, dynamic jaws techniques allow jaws to move during a treatment. On the one hand, the jaws open wide to increase efficiency and thus reduce beam-on time. On the other hand, the jaws can close and follow the target border when sharp penumbra is required near the superior and inferior borders of tumor sites, which results in improved longitudinal dose conformity. The main purpose of the dynamic couch technique is to move the couch as fast as possible at variable speed to reduce beam-on time. Delivering most conformal dose as fast as possible requires a combination of dynamic jaws and dynamic couch techniques (DJDC). Motions of the jaws and couch are determined from the longitudinal fluence profile, which is calculated from an optimized leaf sinogram of small jaw width regular delivery or running start and stop delivery (RSS). We focused on RSS and DJDC in this study and also discussed other delivery techniques. Several conceptual cases are simulated to compare different delivery techniques. The results show that beam-on time can be reduced by about 60% compared to regular delivery with a 2.5 cm jaw width (REG 2.5 cm) for these cases and arbitrary longitudinal fluence profiles can be delivered. Two clinical cases, a prostate and a head-and-neck case, with different delivery techniques are calculated. The results show that plan quality yielded by DJDC with a maximum 5.0 cm jaw width is overall comparable to or better than that of the existing REG 2.5 cm. The DVH comparisons show better critical structure avoidance with the dynamic techniques. At the same time, beam-on time is reduced by about one half compared to REG 2.5 cm. Dynamic delivery techniques provide users more tools to speed up delivery and/or improve plan quality.


Cervical human papillomavirus infection among young women engaged in sex work in Phnom Penh, Cambodia: prevalence, genotypes, risk factors and association with HIV infection.
Although cervical cancer is the leading cancer in Cambodia, most women receive no routine screening for cervical cancer and few treatment options exist. Moreover, nothing is known regarding the prevalence of cervical HPV or the genotypes present among women in the country. Young sexually active women, especially those with multiple sex partners are at highest risk of HPV infection. We examine the prevalence and genotypes of cervical HPV, as well as the associated risk factors among young women engaged in sex work in Phnom Penh, Cambodia. We conducted a cross-sectional study among 220 young women (15-29 years) engaged in sex work in different venues including brothels or entertainment establishments, and on a freelance basis in streets, parks and private apartments. Cervical specimens were collected using standard cytobrush technique. HPV DNA was tested for by polymerase chain reaction (PCR) and genotyping using type-specific probes for 29 individual HPV types, as well as for a mixture of 10 less common HPV types. All participants were also screened for HIV status using blood samples. Multivariate logistic regression analyses were conducted to assess risk factors for any or multiple HPV infection. The prevalence of cervical HPV 41.1%. HPV 51 and 70 were the most common (5.0%), followed by 16 (4.6%), 71 (4.1%) and 81 (3.7%). Thirty-six women (16.4%) were infected with multiple genotypes and 23.3% were infected with at least one oncogenic HPV type. In multivariate analyses, having HIV infection and a higher number of sexual partners were associated with cervical HPV infection. Risk factors for infection with multiple genotypes included working as freelance female sex workers (FSW) or in brothels, recent binge use of drugs, high number of sexual partners, and HIV infection. This is the first Cambodian study on cervical HPV prevalence and genotypes. We found that HPV infection was common among young FSW, especially among women infected with HIV. These results underscore the urgent need for accessible cervical cancer screening and treatment, as well as for a prophylactic vaccine that covers the HPV subtypes present in Cambodia.


Eosinophilic oesophagitis: a common cause of dysphagia in young adults?
Eosinophilic oesophagitis (EO) is an increasingly recognised chronic, relapsing inflammatory condition of the oesophagus. There has been a mini-epidemic of EO in the last decade. The incidence of this condition is higher in children and is commoner in males. There is either a family or personal history of atopic conditions present in a significant number of patients and can also be familial in up to 10%. The classical symptom in an adult is chronic, intermittent solid-food dysphagia and food impaction, often necessitating emergency endoscopic removal. Despite the history of dysphagia for a number of years, patients remain well with no weight loss, which can mislead clinicians to diagnose a functional problem with a resulting delay in the diagnosis. There are various endoscopic features of EO; commonly multiple rings and linear furrows, though these can be subtle and the mucosa may be macroscopically normal. The hallmark of this condition is the histological presence of > or =15 eosinophils/high power field (HPF) in the oesophageal mucosa. Therapeutic options include avoidance of dietary allergens, topical or systemic steroids, Montelukast, Mepolizumab (anti-IL-5 antibody) and endoscopic dilation of strictures unresponsive to medical therapy.


Enhancer-mediated activation of a growth-regulated promoter.
We have demonstrated that the collagen alpha 2 type 1 promoter inserted in an expression vector, behaves as a growth-regulated promoter, which is consistent with previous observations that collagen synthesis is growth-regulated in vivo. In contrast, the activity of the H2-K or the simian virus 40 early promoters does not seem to be affected by the rate of cell proliferation. The insertion of a polyoma enhancer 5' or 3' to the collagen transcription unit activates the collagen alpha 2 type 1 promoter, by a threefold greater factor in slowly growing cells compared to cells growing exponentially. These results show that enhancers can also function in slowly proliferating cells and activate the normally low activity of a promoter in these cells.


Appearance characterization of linear Lambertian objects, generalized photometric stereo, and illumination-invariant face recognition.
Traditional photometric stereo algorithms employ a Lambertian reflectance model with a varying albedo field and involve the appearance of only one object. In this paper, we generalize photometric stereo algorithms to handle all appearances of all objects in a class, in particular the human face class, by making use of the linear Lambertian property. A linear Lambertian object is one which is linearly spanned by a set of basis objects and has a Lambertian surface. The linear property leads to a rank constraint and, consequently, a factorization of an observation matrix that consists of exemplar images of different objects (e.g., faces of different subjects) under different, unknown illuminations. Integrability and symmetry constraints are used to fully recover the subspace bases using a novel linearized algorithm that takes the varying albedo field into account. The effectiveness of the linear Lambertian property is further investigated by using it for the problem of illumination-invariant face recognition using just one image. Attached shadows are incorporated in the model by a careful treatment of the inherent nonlinearity in Lambert's law. This enables us to extend our algorithm to perform face recognition in the presence of multiple illumination sources. Experimental results using standard data sets are presented.


Synthesis and aggregation behaviour of single-chain, 1,32-alkyl branched bis(phosphocholines): effect of lateral chain length.
Three novel single-chain bis(phosphocholines) bearing two lateral alkyl chains of variable length next to the headgroup have been synthesized as model lipids for naturally occurring archaeal membrane lipids. The synthesis was realized using the Cu-catalyzed Grignard bis-coupling reaction of a primary bromide as a side part and a 1,ω-dibromide as a centre part. We could show that the aggregation behaviour of the resulting bolalipids strongly depends on the length of the lateral alkyl chain: the C3-branched bolalipid self-assembles into lamellar sheets, whereas the C6- and C9-analogues form nanofibres. The lamella-forming bolalipids could be used in the future to prepare stable and tailored liposomes for oral drug delivery.


dbPTM in 2019: exploring disease association and cross-talk of post-translational modifications.
The dbPTM (http://dbPTM.mbc.nctu.edu.tw/) has been maintained for over 10 years with the aim to provide functional and structural analyses for post-translational modifications (PTMs). In this update, dbPTM not only integrates more experimentally validated PTMs from available databases and through manual curation of literature but also provides PTM-disease associations based on non-synonymous single nucleotide polymorphisms (nsSNPs). The high-throughput deep sequencing technology has led to a surge in the data generated through analysis of association between SNPs and diseases, both in terms of growth amount and scope. This update thus integrated disease-associated nsSNPs from dbSNP based on genome-wide association studies. The PTM substrate sites located at a specified distance in terms of the amino acids encoded from nsSNPs were deemed to have an association with the involved diseases. In recent years, increasing evidence for crosstalk between PTMs has been reported. Although mass spectrometry-based proteomics has substantially improved our knowledge about substrate site specificity of single PTMs, the fact that the crosstalk of combinatorial PTMs may act in concert with the regulation of protein function and activity is neglected. Because of the relatively limited information about concurrent frequency and functional relevance of PTM crosstalk, in this update, the PTM sites neighboring other PTM sites in a specified window length were subjected to motif discovery and functional enrichment analysis. This update highlights the current challenges in PTM crosstalk investigation and breaks the bottleneck of how proteomics may contribute to understanding PTM codes, revealing the next level of data complexity and proteomic limitation in prospective PTM research.


Molecular relationships between Australian annual wild rice, Oryza meridionalis, and two related perennial forms.
The perennial, Oryza rufipogon distributed from Asia to Australia and the annual O. meridionalis indigenous to Australia are AA genome species in the Oryza. However, recent research has demonstrated that the Australian AA genome perennial populations have maternal genomes more closely related to those of O. meridionalis than to those found in Asian populations of O. rufipogon suggesting that the Australian perennials may represent a new distinct gene pool for rice. Analysis of an Oryza core collection covering AA genome species from Asia to Oceania revealed that some Oceania perennials had organellar genomes closely related to that of O meridionalis (meridionalis-type). O. rufipogon accessions from New Guinea carried either the meridionalis-type or rufirpogon-type (like O. rufipogon) organellar genomes. Australian perennials carried only the meridionalis-type organellar genomes when accompanied by the rufipogon-type nuclear genome. New accessions were collected to better characterize the Australian perennials, and their life histories (annual or perennial) were confirmed by field observations. All of the material collected carried only meridionalis-type organellar genomes. However, there were two distinct perennial groups. One of them carried an rufipogon-type nuclear genome similar to the Australian O. rufipogon in the core collection and the other carried an meridionalis-type nuclear genome not represented in the existing collection. Morphologically the rufipogon-type shared similarity with Asian O. rufipogon. The meridionalis-type showed some similarities to O. meridionalis such as the short anthers usually characteristic of annual populations. However, the meridionalis-type perennial was readily distinguished from O. meridionalis by the presence of a larger lemma and higher number of spikelets. Analysis of current accessions clearly indicated that there are two distinct types of Australian perennials. Both of them differed genetically from Asian O. rufipogon. One lineage is closely related to O. meridionalis and another to Asian O. rufipogon. The first was presumed to have evolved by divergence from O. meridionalis becoming differentiated as a perennial species in Australia indicating that it represents a new gene pool. The second, apparently derived from Asian O. rufipogon, possibly arrived in Australia later.


Effect of myo-inositol and melatonin versus myo-inositol, in a randomized controlled trial, for improving in vitro fertilization of patients with polycystic ovarian syndrome.
Polycystic ovarian syndrome (PCOS) induces anovulation in women of reproductive age, and is one of the pathological factors involved in the failure of in vitro fertilization (IVF). Indeed, PCOS women are characterized by poor quality oocytes. Therefore, a treatment for enhancing oocyte quality becomes crucial for these patients. Myo-Inositol and melatonin proved to be efficient predictors for positive IVF outcomes, correlating with high oocyte quality. We tested the synergistic effect of myo-inositol and melatonin in IVF protocols with PCOS patients in a randomized, controlled, double-blind trial. Five-hundred twenty-six PCOS women were divided into three groups: Controls (only folic acid: 400 mcg), Group A (Inofolic® plus, a daily dose of myo-inositol: 4000 mg, folic acid: 400 mcg, and melatonin: 3 mg), and Group B (Inofolic®, a daily dose of myo-inositol: 4000 mg, and folic acid: 400 mcg). The main outcome measures were oocyte and embryo quality, clinical pregnancy and implantation rates. The treatment lasted from the first day of the cycle until 14 days after embryo transfer. Myo-inositol and melatonin have shown to enhance, synergistically, oocyte and embryo quality. In consideration of the beneficial effect observed in our trial and on the bases of previous studies, we decided to integrate routinely MI and M supplementation in the IVF protocols. The same treatment should be taken carefully in consideration in all procedures of this kind.


Whole-Cell Properties of Cerebellar Nuclei Neurons In Vivo.
Cerebellar nuclei neurons integrate sensorimotor information and form the final output of the cerebellum, projecting to premotor brainstem targets. This implies that, in contrast to specialized neurons and interneurons in cortical regions, neurons within the nuclei encode and integrate complex information that is most likely reflected in a large variation of intrinsic membrane properties and integrative capacities of individual neurons. Yet, whether this large variation in properties is reflected in a heterogeneous physiological cell population of cerebellar nuclei neurons with well or poorly defined cell types remains to be determined. Indeed, the cell electrophysiological properties of cerebellar nuclei neurons have been identified in vitro in young rodents, but whether these properties are similar to the in vivo adult situation has not been shown. In this comprehensive study we present and compare the in vivo properties of 144 cerebellar nuclei neurons in adult ketamine-xylazine anesthetized mice. We found regularly firing (N = 88) and spontaneously bursting (N = 56) neurons. Membrane-resistance, capacitance, spike half-width and firing frequency all widely varied as a continuum, ranging from 9.63 to 3352.1 MΩ, from 6.7 to 772.57 pF, from 0.178 to 1.98 ms, and from 0 to 176.6 Hz, respectively. At the same time, several of these parameters were correlated with each other. Capacitance decreased with membrane resistance (R2 = 0.12, P<0.001), intensity of rebound spiking increased with membrane resistance (for 100 ms duration R2 = 0.1503, P = 0.0011), membrane resistance decreased with membrane time constant (R2 = 0.045, P = 0.031) and increased with spike half-width (R2 = 0.023, P<0.001), while capacitance increased with firing frequency (R2 = 0.29, P<0.001). However, classes of neuron subtypes could not be identified using merely k-clustering of their intrinsic firing properties and/or integrative properties following activation of their Purkinje cell input. Instead, using whole-cell parameters in combination with morphological criteria revealed by intracellular labelling with Neurobiotin (N = 18) allowed for electrophysiological identification of larger (29.3-50 μm soma diameter) and smaller (< 21.2 μm) cerebellar nuclei neurons with significant differences in membrane properties. Larger cells had a lower membrane resistance and a shorter spike, with a tendency for higher capacitance. Thus, in general cerebellar nuclei neurons appear to offer a rich and wide continuum of physiological properties that stand in contrast to neurons in most cortical regions such as those of the cerebral and cerebellar cortex, in which different classes of neurons operate in a narrower territory of electrophysiological parameter space. The current dataset will help computational modelers of the cerebellar nuclei to update and improve their cerebellar motor learning and performance models by incorporating the large variation of the in vivo properties of cerebellar nuclei neurons. The cellular complexity of cerebellar nuclei neurons may endow the nuclei to perform the intricate computations required for sensorimotor coordination.


Kawasaki disease presenting as cervical lymphadenitis or deep neck infection.
To describe a group of patients with Kawasaki disease who had cervical lymphadenopathy as their dominant initial presentations. We retrospectively reviewed the medical records of 14 children who were admitted to Chang-Gung Children's Hospital between May 1996 and July 1998 with the initial impression of cervical lymphadenitis, cellulitis, and/or deep neck infection but for which a diagnosis of Kawasaki disease was established later. Five (35.7%) patients were less than 5 months of age, and 8 (57.1%) patients were more than 53 months of age. The mean duration for establishing a diagnosis of Kawasaki disease from the onset of illness was 8.2 (6 to 20) days. Initially, empiric antibiotics were prescribed in each case with unsatisfactory response. Intravenous immune gamma globulin (2 g/kg) was administered in 13 patients. Three (21.4%) patients developed coronary artery lesions. If a child less than 6 months or more than 4 years of age has a fever and an enlarged cervical lymph node and is unresponsive to empiric antibiotics, Kawasaki disease should be considered.


A free boundary mechanobiological model of epithelial tissues.
In this study, we couple intracellular signalling and cell-based mechanical properties to develop a novel free boundary mechanobiological model of epithelial tissue dynamics. Mechanobiological coupling is introduced at the cell level in a discrete modelling framework, and new reaction-diffusion equations are derived to describe tissue-level outcomes. The free boundary evolves as a result of the underlying biological mechanisms included in the discrete model. To demonstrate the accuracy of the continuum model, we compare numerical solutions of the discrete and continuum models for two different signalling pathways. First, we study the Rac-Rho pathway where cell- and tissue-level mechanics are directly related to intracellular signalling. Second, we study an activator-inhibitor system which gives rise to spatial and temporal patterning related to Turing patterns. In all cases, the continuum model and free boundary condition accurately reflect the cell-level processes included in the discrete model.