Cloning and sequencing of kojibiose phosphorylase gene from Thermoanaerobacter brockii ATCC35047.
A gene encoding kojibiose phosphorylase was cloned from Thermoanaerobacter brockii ATCC35047. The kojP gene encodes a polypeptide of 775 amino acid residues. The deduced amino acid sequence was homologous to those of trehalose phosphorylase from T. brockii and maltose phosphorylases from Bacillus sp. and Lactobacillus brevis with 35%, 29% and 28% identities, respectively. Kojibiose phosphorylase was efficiently overexpressed in Escherichia coli JM109. The DNA sequence of 3956 bp analyzed in this study contains three open reading frames (ORFs) downstream of kojP. The four ORFs, kojP, kojE, kojF, and kojG, form a gene cluster. The amino acid sequences deduced from kojE and kojF are similar to those of the N-terminal and C-terminal regions of a sugar-binding periplasmic protein from Thermoanaerobacter tengcongensis MB4. Furthermore, the amino acid sequence deduced from kojG is similar to that of a permease of the ABC-type sugar transport systems from T. tengcongensis MB4. Each of three amino acid substitutions, D362N, K614Q and E642Q, caused a complete loss of kojibiose phosphorylase activity. These results suggest that D362, K614 and E642 play an important role in catalysis. Another mutation, D459N, increased K(m) values for kojibiose (7-fold that for the wild type), beta-G1P (11-fold) and glucose (7-fold), whereas K(m) for inorganic phosphate was minimally affected by this mutation, suggesting that D459 may be involved in the binding to saccharides.


Supersensitive Oxidation-Responsive Biodegradable PEG Hydrogels for Glucose-Triggered Insulin Delivery.
Reactive oxygen species (ROS)-responsive polymers and hydrogels represent an emerging family of intelligent materials owing to the key functions of ROS in physiological processes or pathological diseases. Nonetheless, the weaknesses such as low sensitivity, slow response, instability, and low mechanical strength are associated with the limited ROS-responsive polymeric or supramolecular hydrogels. In this study, a novel type of oxidation-responsive degradable hydrogels was fabricated by the redox-initiated radical polymerization of a 4-arm-poly(ethylene glycol) (PEG) acrylic macromonomer that possesses a H2O2-cleavable phenylboronic acid linker in each of the arms. The macroscopic hydrogels have the features of good cytocompatibility, moderate mechanical strength, and fast response toward H2O2 of low concentration, owing to the covalently cross-linked hydrophilic PEG network and high sensitivity of the linker. They could encapsulate biomacromolecules, such as insulin and glucose oxidase (GOx), with high efficacy, affording a new glucose-responsive insulin-delivery platform on the basis of enzymatic transformation of a biochemical signal (glucose) into an oxidative stimulus (H2O2). Interestingly, in vitro results demonstrate that the same GOx-loaded hydrogel exhibited disparate degradation modes under different triggering molecules, that is, bulk degradation by H2O2 and surface erosion by glucose. Moreover, compared to the macroscopic hydrogel, the nanogel with a diameter of ∼160 nm prepared by inverse emulsion polymerization showed a much higher degradation rate even under triggering of 20 μM H2O2, a pathologically available concentration in vivo.


How midwives implemented teleconsultations during the COVID-19 health crisis: a mixed-methods study.
Our primary and secondary objectives were to measure and understand the determinants of independent midwives' implementation of teleconsultations and their intention to continue these in the future. A two-phase mixed-methods approach: (1) quantitative data to measure factors determining the initiation and continuation of teleconsultation, collected by an online survey from 29 April to 15 May 2020, at the end of the first COVID-19 lockdown, followed by (2) qualitative data to understand these determinants, by interviewing some participants in May-July 2020 to explore the quantitative findings in more detail. Mainland France PARTICIPANTS: The target population comprised independent midwives currently practising in France. The primary and secondary outcomes were binary variables: implementation of teleconsultations, and intention to continue them. The qualitative results provided the themes explaining these decisions. We obtained 1491 complete responses from independent midwives, that is, 28.3% of French independent midwives, and interviewed 22 volunteers among them. Among the 1491, 88.5% implemented teleconsultations and 65.8% intended to continue them. Both individual and organisational factors favoured implementation of teleconsultations: older age (adjusted OR (aOR): 0.40, 95% CI: 0.28 to 0.58), female gender (aOR: 6.88, 95% CI: 2.71 to 17.48), married or living with a partner (aOR: 1.67, 95% CI: 1.10 to 2.52) and working in a group practice (midwives only-aOR: 2.34, 95% CI: 1.47 to 3.72; multiprofessional group-aOR: 1.75, 95% CI: 1.16 to 2.64). The qualitative analysis did not identify any new factors but helped us to understand the satisfaction better: midwives adopted telemedicine for their patients' access to and continuity of care, to maintain their professional activity and income, and to limit the risks of infection. Personal and organisational factors motivated the implementation of teleconsultation during the pandemic, but maintaining it raises technical, regulatory, and ethical issues.


Combined kinase inhibition modulates parkin inactivation.
Mutations in the parkin gene cause autosomal-recessive, juvenile-onset parkinsonism, and parkin dysfunction may also play a role in the pathogenesis of sporadic Parkinson disease (PD). Although its precise function remains largely unknown, parkin seems to play a neuroprotective role. Several studies indicate that changes in parkin solubility induced by post-translational modifications, such as S-nitrosylation or dopamine modification, comprise one mechanism of parkin inactivation associated with disease. Protein phosphorylation events have recently been linked to the molecular mechanism(s) underlying PD, but the role of this post-translational modification for parkin function has remained unclear. Here we report that compound phosphorylation of parkin by both casein kinase I and cyclin-dependent kinase 5 (cdk5) decreases parkin solubility, leading to its aggregation and inactivation. Combined kinase inhibition partially reverses the aggregative properties of several pathogenic point mutants in cultured cells. Enhanced parkin phosphorylation is detected in distinct brain areas of individuals with sporadic PD and correlates with increases in the levels of p25, the activator of cdk5. These findings indicate that casein kinase I and cdk5 may represent novel combinatorial therapeutic targets for treating PD.


Extended FTLD pedigree segregating a Belgian GRN-null mutation: neuropathological heterogeneity in one family.
In this paper, we describe the clinical and neuropathological findings of nine members of the Belgian progranulin gene (GRN) founder family. In this family, the loss-of-function mutation IVS1 + 5G > C was identified in 2006. In 2007, a clinical description of the mutation carriers was published that revealed the clinical heterogeneity among IVS1 + 5G > C carriers. We report our comparison of our data with the published clinical and neuropathological characteristics of other GRN mutations as well as other frontotemporal lobar degeneration (FTLD) syndromes, and we present a review of the literature. For each case, standardized sampling and staining were performed to identify proteinopathies, cerebrovascular disease, and hippocampal sclerosis. The neuropathological substrate in the studied family was compatible in all cases with transactive response DNA-binding protein (TDP) proteinopathy type A, as expected. Additionally, most of the cases presented also with primary age-related tauopathy (PART) or mild Alzheimer's disease (AD) neuropathological changes, and one case had extensive Lewy body pathology. An additional finding was the presence of cerebral small vessel changes in every patient in this family. Our data show not only that the IVS1 + 5G > C mutation has an exclusive association with FTLD-TDP type A proteinopathy but also that other proteinopathies can occur and should be looked for. Because the penetrance rate of the clinical phenotype of carriers of GRN mutations is age-dependent, further research is required to investigate the role of co-occurring age-related pathologies such as AD, PART, and cerebral small vessel disease.


Validation of a diffusive sampling method for airborne low-molecular isocyanates using 4-nitro-7-piperazinobenzo-2-oxa-1,3-diazole-impregnated filters and liquid chromatography-tandem mass spectrometry.
A diffusive sampling method for the determination of low-molecular isocyanates as their 4-nitro-7-piperazinobenzo-2-oxa-1,3-diazole (NBDPZ) derivatives using tandem mass spectrometry (MS/MS) after atmospheric pressure chemical ionisation (APCI) is presented. Isocyanic acid (ICA), methyl isocyanate (MIC), ethyl isocyanate (EIC) and phenyl isocyanate (PhIC) are collected on NBDPZ-impregnated polystyrene divinyl benzene (SDB) filter tapes. The method was validated for MIC, EIC and PhIC for concentrations between 0.5 and 50 ppb at relative humidity (RH) conditions from 10 up to 90%. Validation was carried out by active sampling using 1-(2-methoxyphenyl)piperazine (2-MP) as derivatising agent. Sampling periods applied were between 15 min and more than 8 h. The sampling rates were determined to be 21.0 mL/min for MIC with a relative standard deviation (RSD) of 9.0% for 184 samplers, 15.6 mL/min for EIC (RSD 11.6%; N = 154) and 11.5 mL/min for PhIC (RSD 8.4%; N = 87). The limits of quantification were 1.4 ppb for MIC and 1.3 ppb for EIC and PhIC applying 15 min sampling periods. Owing to high background signals, isocyanic acid could only be determined when it was present in concentrations in the high ppb range.


Molecular characterization of NOS in a mollusc: expression in a giant modulatory neuron.
Here we report on the molecular characterization of the first molluscan NOS in the CNS of the pond snail Lymnaea stagnalis. This Lymnaea NOS (Lym-nNOS) which is expressed preferentially in the CNS is most similar to mammalian neuronal NOS but contains tandem repeats of a seven amino acid motif not found in any other known NOS. We have localized Lym-nNOS to the serotonergic cerebral giant cells (CGCs) which modulate synaptic transmission within a neural network that generates feeding behavior. Our results suggest that the CGCs employ both NO and serotonin in the modulation of the central neural network underlying feeding.


Current concepts on bronchoalveolar lavage cells in extrinsic allergic alveolitis.
Studies performed on cells recovered from bronchoalveolar lavage fluid and lung tissue sections have demonstrated that the alveolitis in patients with extrinsic allergic alveolitis is characterized by cells bearing suppressor/cytotoxic phenotype (T3+, T8+, HNK1+). Functional in vitro analyses demonstrated that lung T cells from these patients are able to provide suppressor function in a pokeweed-mitogen-induced B cell differentiation assay and nonspecific cytotoxic activity. This latter property has not been observed in subjects with asymptomatic extrinsic allergic alveolitis. The expansion of cells with suppressor and cytotoxic characteristics in the lung of these patients is likely to be related to a local immunologic response to the antigenic stimulus.


Primary structure analysis and adhesion studies on the major outer membrane protein of Campylobacter jejuni.
The major outer membrane protein of Campylobacter jejuni (MOMP, 43 kDa), supposed to be one of the structures responsible for adhesion to INT 407 cells, was isolated from the crude outer membrane preparation by treatment with n-octyl-beta-D-glucopyranoside followed by preparative SDS-polyacrylamide gel electrophoresis. By cleavage of the isolated protein with cyanogen bromide and proteolytic enzymes, peptides were generated, separated by reverse phase high pressure liquid chromatography, and sequenced by automatic Edman degradation. The protein was aligned by identification of overlapping peptides. Treatment of bacteria with proteinase K prior to preparation of the outer membrane yielded a truncated MOMP with an apparent molecular mass of 25 kDa consisting of the C-terminal part of the protein. The isolated MOMP was functionally characterized by significant binding activity towards INT 407 cell membranes when isolated by preparative native gel electrophoresis, however, no binding activity was detected when the protein was isolated in the presence of SDS.


Somatoform Dissociation, Fatigue Severity and Pain Behavior Compared in Patients with Migraine Headache and in Healthy Individuals.
The prevalence of migraine in the world is about 15 and 7% among women and men, respectively. The purpose of this study was comparison of somatoform dissociation, fatigue severity and pain behavior in patients with migraine headache and its relationship with coping strategies. This descriptive analytical study has been done on 120 patients with migraine headache and 120 healthy subjects were selected randomly. Data collected by somatoform dissociation questionnaire (SDQ-20), fatigue severity scale, pain behavior scale and coping strategies scale. For data analysis we used SPSS.19. The means of the somatoform dissociation, pain behavior scale, help searching subscale and pain compliant in migraine and healthy subjects were statistically significant. There was not significant difference in avoidance subscales between the two groups. Comparison of fatigue severity in patients with migraine and control group was meaningful. There was significant positive correlation between all four scales and coping strategies. It seems that these symptoms can play an important role in this disease; thus, their careful evaluation in the treatment of migraine headache is essential.


Impairment of Macrophage Presenting Ability and Viability by Echinococcus granulosus Antigens.
Despite advances toward an improved understanding of the evasive mechanisms leading to the establishment of cystic echinococcosis, the discovery of specific immunosuppressive mechanisms and related factors are of great interest in the development of an immunotherapeutic approach. To elucidate immunosuppressive effects of bioactive factors contained in chromatographic fractions from hydatid cystic fluid (HCF) of Echinococcus granulosus. Hydatid cystic fluid was fractionated by reverse phase chromatography. Non-specific Concanavalin A-driven proliferation of spleen cells was used to determine specific inhibitory fractions. Trypan blue exclusion test and flowcytometry analysis were performed to check whether highly inhibitory fractions of HCF have apoptotic effect on peritoneal macrophages. Western blot analysis was used to determine proteolytic effects of parasitic antigens on major histocompatibility complex (MHC) class II (I-a) contained in membrane proteins extract from macrophages. High concentrations of HCF and few of chromatographic fractions suppressed spleen cells proliferation. Fractions 7 and 35 were the highest inhibitory fractions. Specifically fraction 35 and to a lesser extent HCF induced apoptosis in peritoneal naive macrophages. However, HCF and the fraction 7 proteolytically altered the expression of MHC class II molecules on peritoneal macrophages. The proteolytic molecule was identified to be a serine protease. Macrophages taken at the chronic and end phase from cystic echinococcosis-infected mice were able to uptake and process C-Ovalbumine-FITC. These cells expressed a drastically reduced level of (I-a) molecules. Our study present new aspects of immune suppression function of E. granulosus. Further molecular characterization of apoptotic and proteolytic factors might be useful to develop immunotherapeutic procedure to break down their inhibitory effects.


Purification of a Ran-interacting protein that is required for protein import into the nucleus.
Previously we reported the isolation of two cytosolic fractions (A and B) from Xenopus ovary that are required sequentially to support protein import into the nuclei of digitonin-permeabilized cells. Fraction A is required for recognition of the nuclear localization sequence and targeting to the nuclear envelope, whereas fraction B is required for the subsequent translocation of the bound substrate into the nucleus. The first protein required for fraction B activity to be purified was the small GTPase Ran (ras-related nuclear protein). Here we report the purification of the second (and final) protein required for fraction B activity. By SDS/PAGE, the purified protein appeared as a single band with an apparent molecular mass of 10 kDa, but the native protein fractionated upon gel filtration chromatography with an apparent size of 30 kDa. Peptide sequence analysis indicated that the purified protein was highly homologous to a previously identified human protein of unknown function called placental protein 15 (pp15) and to the predicted protein product of a yeast open reading frame from Saccharomyces cerevisiae.


Association between vitamin B12 levels and cognitive impairment in older adults.
Whether vitamin B12 deficiency is associated with cognitive impairment remains controversial. To determine the association between vitamin B12 serum levels and cognitive performance. Two-hundred and forty-one adults aged ≥ 60 years who had serum vitamin B12 serum levels measurement were included. Physical and cognitive evaluation was carried out, and three groups were formed: normal cognition (NC), mild cognitive impairment (MCI) and dementia. Vitamin B12 levels were classified as sufficiency (> 400 pg/mL), subclinical deficiency (201-400 pg/mL), and absolute deficiency (≤ 200 pg/mL). Multivariate linear regression analysis was used to evaluate the association between cognitive function and vitamin B12 levels after controlling for confounding variables. Mean age was 81.4 ± 8.0 years; 68% were females; 17.8 % and 39.8% had absolute and subclinical vitamin B12 deficiency, respectively; 80 individuals (33%) met the criteria for MCI, and 70 (29%), for dementia. Those with MCI and dementia had lower vitamin B12 levels in comparison with those with NC after adjusting for age, gender and educational level (p = 0.019). A statistically significant association was observed between global cognitive performance and levels of vitamin B12.


Keratinization of palatal mucosa beneath metal-based removable partial and acrylic-based complete dentures compared with normal palatal mucosa: a clinical, cytological and histological study.
Normal human palatal epithelium is an orthokeratinizing tissue with a well-developed stratum corneum. Several cytological and histological studies showed that when the palate covered by a denture fully or partially, keratinization of the mucosa changes. The results of some studies suggested that keratinization changes from orthokeratosis to parakeratosis, and there is a reduction in the thickness of stratum corneum. Whereas the results of some other studies showed increased orthokeratinization. This study was planned to determine changes in keratinization of palatal mucosa in denture wearers compared with normal palatal mucosa. Keratinization of palatal mucosa was examined by mainly exfoliative cytology in three groups. Group-1 consisted of 51 patients wearing metal-based removable partial denture; group-2 consisted of 50 patients wearing acrylic-based complete denture and group-3 including 57 healthy volunteers who have not been wearing denture served as the controls. In each group, the results of exfoliative cytology were supported by histological examinations. Statistical results showed that either acrylic-based complete or cobalt-chromium-based removable partial dentures seem to reduce the quantity and/or quality of the keratin layer. The mean values of keratinization and the thickness of stratum corneum were significantly lower in the complete denture group than in the partial denture group. Furthermore the reliability of exfoliative cytology was confirmed by the guidance of histological examination.


Involvement of upper-airway muscles in extrapyramidal disorders. A cause of airflow limitation.
To identify the site and cause of airflow limitation in patients with parkinsonism, we tested pulmonary function in 27 patients with extrapyramidal disorders. In 24 patients, an abnormal flow-volume loop contour, showing either regular (18 patients) or irregular (6 patients) flow oscillations, was found. On direct fiberoptic visualization of the upper airway, these oscillations corresponded to either rhythmic (4 to 8 Hz) or irregular involuntary movements of glottic and supraglottic structures. Ten patients had physiologic evidence of upper-airway obstruction, which was symptomatic in four. We conclude that the upper-airway musculature is frequently involved in extrapyramidal disorders. This causes upper-airway dysfunction that can be severe enough to limit airflow.


An item response theory analysis of the sexual compulsivity scale and its correspondence with the hypersexual disorder screening inventory among a sample of highly sexually active gay and bisexual men.
Numerous scales and assessments are available to assess sexual compulsivity (SC). This study sought to conduct an item response theory (IRT) analysis of the Sexual Compulsivity Scale (SCS) to provide evidence about its measurement precision at the various levels of the SC construct in a sample of highly sexually active gay and bisexual men (GBM). SCS data from a sample of 202 GBM who are highly sexually active but who vary in their experiences of SC symptoms were modeled using Samejima's polytomous graded response IRT model. To describe the performance of the SCS relative to the Hypersexual Disorder Screening Inventory (HDSI), SCS scores were compared with participants' corresponding HDSI results to determine sensitivity, specificity, positive and negative predictive values, and accuracy. This study examined the correspondence between the SCS and the HDSI, a diagnostic instrument for the screening of hypersexuality. IRT analyses indicated that, although two of the SCS items had low reliability, the SCS as a whole was reliable across much of the SC continuum. Scores on the SCS and the HDSI were highly correlated; however, no potential cutoffs on the SCS corresponded strongly with the polythetic scoring criteria of the HDSI. Comparisons of SCS scores with HDSI results indicated that the SCS itself could not serve as a substitute for the HDSI and would incorrectly classify a substantial number of individuals' levels of hypersexuality. However, the SCS could be a useful screening tool to provide a preliminary screening of people at risk for meeting criteria on the HDSI. Combining the SCS and the HDSI may be an appropriate evaluation strategy in classifying GBM as negative on both (i.e., "non-hypersexual/non-SC"), positive on the SCS only (i.e., "at risk"), and positive on both the SCS and the HDSI (i.e., "problematic hypersexuality/SC").


Examination of immune parameters and host resistance mechanisms in B6C3F1 mice following adult exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin.
Adult female B6C3F1 mice were given a single ip dose of 0, 01, 1.0, or 10.0 micrograms/kg 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and examined for immune function and host resistance 7-10 d later. Exposure to TCDD resulted in a significant dose-related decrease in induction of both IgM and IgG antibody-forming cells. This suppression was noted for both T-dependent and T-independent antigens. TCDD at a dosage of 10 micrograms/kg was shown to suppress production of antibody to viral hemagglutinin. In contrast, TCDD exposure had no significant effect on natural killer cell function, production of interferon, or various parameters of macrophage function. Assessment of host resistance revealed a significant increase in susceptibility to fatal infection with influenza virus, but no significant alteration in susceptibility to infection with the bacterium Listeria monocytogenes.


Vibration and acoustic response of an orthotropic composite laminated plate in a hygroscopic environment.
This paper is a study of the vibration and acoustic response characteristics of orthotropic laminated composite plate with simple supported boundary conditions excited by a harmonic concentrated force in a hygroscopic environment. First the natural vibration of the plate with the in-plane forces induced by hygroscopic stress is obtained analytically. Secondly, the sound pressure distribution of the plate at the far field is obtained using the Rayleigh integral. Furthermore, the sound radiation efficiency is deduced. Third, different ratios of elastic modulus in material principal directions are set to research the effects of increasing stiffness of the orthotropic plate on the vibration and acoustic radiation characteristics. Finally, to verify the theoretical solution, numerical simulations are also carried out with commercial finite software. It is found that the natural frequencies decrease with the increase of the moisture content and the first two order modes interconvert at high moisture content. The dynamic response and sound pressure level float to lower frequencies with elevated moisture content. Acoustic radiation efficiency generally floats to the low frequencies and decreases with an increase of moisture content. The dynamic and acoustic responses reduce and the coincidence frequency decreases with the enhanced stiffness.


IQ-Achievement Discrepancy for Identification of Disabilities in Spanish-speaking English Learners.
This article examines the validity of IQ-achievement discrepancy and low achievement as criteria for the identification of disabilities in Spanish-speaking English-language learners (ELs) and the factors that moderate the validity of these approaches as bases for identification. While there has been a long history of examining the validity of different approaches to disability identification in monolinguals, there are no systematic approaches taken for ELs. Data from Grades 1 and 2 of a large longitudinal data set consisting of young Spanish-speaking students attending schools in the United States were used to empirically examine criteria for disability identification among language minority children-one of the first large-scale attempts. Findings indicated significant overidentification when the language of assessment was not matched to the language of the instruction, although the effects varied predictably over time and by language of instruction. Validation of classifications using measures external to the classification found that low achieving and discrepant children differ from typically developing children, and from one another in predictable ways based on differences in IQ. The study highlights the importance of taking into account the language of instruction and the severity of the cut-off to reduce misidentification of typically developing children.


M1 muscarinic agonist treatment reverses cognitive and cholinergic impairments of apolipoprotein E-deficient mice.
Recent studies suggest that apolipoprotein E (apoE) plays a specific role in brain cholinergic function and that the E4 allele of apoE (apoE4), a major risk factor for Alzheimer's disease (AD), may predict the extent of cholinergic dysfunction and the efficacy of cholinergic therapy in this disease. Animal model studies relevant to this hypothesis revealed that apoE-deficient (knockout) mice have working memory impairments that are associated with distinct dysfunction of basal forebrain cholinergic neurons. Cholinergic replacement therapy utilizing M1-selective muscarinic agonists has been proposed as effective treatment for AD patients. In the present study, we examined whether the memory deficits and brain cholinergic deficiency of apoE-deficient mice can be ameliorated by the M1-selective agonist 1-methylpiperidine-4-spiro-(2'-methylthiazoline), [AF150(S)]. Treatment of apoE-deficient mice with AF150(S) for 3 weeks completely abolished their working memory impairments. Furthermore, this reversal of cognitive deficit was associated with a parallel increase of histochemically determined brain choline acetyltransferase and acetylcholinesterase levels and with the recovery of these cholinergic markers back to control levels. These findings show that apoE deficiency-related cognitive and cholinergic deficits can be ameliorated by M1-selective muscarinic treatment. They also provide a novel model system for development and evaluation of therapeutic strategies directed specifically at the AD patients whose condition is attributed to the apoE genotype.


Rural rotations for interns: a demonstration programme in South Australia.
The Commonwealth Government of Australia, through policy initiatives and increased funding, has placed significant emphasis on increasing undergraduate rural experiences for medical students. However, in the immediate postgraduate years, rural community based rotations are uncommon, with the vast majority of intern experiences remaining hospital based. Since 1997, Flinders Medical Centre, Flinders University and the rural communities of Cleve and Jamestown have run a preregistration rural intern-training position based in rural general practice. The present article describes the programme, its evolution and the problems that have been overcome. The term provides a blend of hospital and community based experiences appropriate for junior doctors not yet familiar with ambulatory care. At the same time, the junior doctors have consistently reported a high-quality learning experience, with ready access to patients and procedural work. We describe the qualitative and quantitative methods we have recently introduced for evaluation of the programme.


[Scanning electron microscopic study of keratohyaline granules in the oral epithelium of rats].
Scanning electron microscopy of rat buccal epithelium on deparaffinized or epon-etched sections provided observations of intracellular structures especially of keratohyaline granules. Cell-walls and nuclei were easily observable and the two types of keratohyaline granules already seen in transmission electron microscopy were identified namely intracytoplasmic or intranuclear single granules and composite granules with a heterogeneous structure which appeared clearly in the scanning method. Tridimensional information from scanning electron microscopy adds significantly to knowledge of keratohyaline granules morphology, so far studied with the transmission method only.


[Heteropolycyclic systems. V. 4-alkyl-2,3,3a,4-tetrahydro-1H-pyrro1/2,1-c/ /1,2,4/benzothiadiazin-5,5-dioxide and their bz-substituted derivatives].
The preparation of some 4-alkyl derivatives of 2,3,3a,4--tetrahydro-1H-pyrrolo(2,1-c)(1,2,4)benzothiadiazin-5,5-dioxode and bz-substituted derivatives by the action of alkyl iodides on the LiAlH4 reduction products of the corresponding 2,3-dihydro-1H-pyrrolo(2,1-c)(1,2,4)benzothiadiazin-5,5-dioxide is described. It was found that some of the substances synthesized exist in polymorphous forms and it was possible to isolate these.


The mechanism of sorbitol dehydration in hot acidic solutions.
Sugar alcohol dehydration in hot water is an important reaction that allows for environmentally friendly biomass conversion without the use of organic solvents. Here, we report a free-energy analysis by metadynamics (MTD) simulations based on ab initio density functional theory and semiempirical density-functional tight-binding method to understand the mechanism of dehydration reactions of d-sorbitol (SBT) in hot acidic water. Comparing the results of ab initio and semiempirical MTD, it was found that the latter gives a reliable free energy surface of SBT dehydration reaction, although the results vary upon the inclusion of dispersion correction. It was found that the reaction proceeds consistently via an SN 2 mechanism, whereby the free energy of protonation of the hydroxyl group created as an intermediate is affected by the acidic species. This mechanism was further verified by real-time trajectories started from the transition state using ab initio molecular dynamics simulations. The free energy barriers of the reaction pathways leading to five-membered ether products are lower than those leading to six-membered ether products, in agreement with experiment. This outcome can be ascribed, in part, to our finding that the reaction barrier of the pathway is correlated to the stability of the SBT confined conformation at the initial stage of the reaction.


Photoinitiated Dynamics in Amorphous Solid Water via Nanoimprint Lithography.
Laser pulses that act on fragile samples often alter them irreversibly, motivating single-pulse data collection. Amorphous solid water (ASW) is a good example. In addition, neither well-defined paths for molecules to travel through ASW nor sufficiently small samples to enable molecular dynamics modeling have been achieved. Combining nanoimprint lithography and photoinitiation overcomes these obstacles. An array of gold nanoparticles absorbs pulsed (10 ns) 532 nm radiation and converts it to heat, and doped ASW films grown at about 100 K are ejected from atop the irradiated nanoparticles into vacuum. The nanoparticles are spaced from one another by sufficient distance that each acts independently. Thus, a temporal profile of ejected material is the sum of about 106 "nanoexperiments," yielding high single-pulse signal-to-noise ratios. The size of a single nanoparticle and its immediate surroundings is sufficiently small to enable modeling and simulation at the atomistic (molecular) level, which has not been feasible previously. An application to a chemical system is presented in which H/D scrambling is used to infer the presence of protons in films composed of D2O and H2O (each containing a small amount of HDO contaminant) upon which a small amount of NO2 has been deposited. The pulsed laser heating of the nanoparticles promotes NO2/N2O4 hydrolysis to nitric acid, whose protons enhance H/D scrambling dramatically.


Near infrared muscle spectroscopy in patients with Friedreich's ataxia.
Friedreich's ataxia is a progressive neurodegenerative disorder of the afferent cerebellar pathways associated with mitochondrial dysfunction at the cellular level. We have used noninvasive continuous near infrared muscle spectroscopy (NIRS) to investigate the delivery and utilization of oxygen in response to exercise in this disorder. Patients performed an incremental treadmill walking protocol in which levels of muscle deoxygenation or oxygenation were continuously measured in the medial calf muscle. The kinetics of recovery from exercise-induced deoxygenation, called the half-time of recovery (t(1/2)) were determined. The t(1/2) was prolonged in patients with Friedreich's ataxia compared with controls, and the degree of prolongation correlated with the length of the shorter GAA repeat, a genetic measure that correlates with the age of onset of disease. The t(1/2) also correlated inversely with patient age and with the maximum treadmill speed attained. Several patients also displayed features consistent with inadequate oxygen utilization by muscle. These results suggest that NIRS may be an effective tool for monitoring the biochemical and functional features of Friedreich's ataxia in parallel.


Water oxidation catalysis: electrocatalytic response to metal stoichiometry in amorphous metal oxide films containing iron, cobalt, and nickel.
Photochemical metal-organic deposition (PMOD) was used to prepare amorphous metal oxide films containing specific concentrations of iron, cobalt, and nickel to study how metal composition affects heterogeneous electrocatalytic water oxidation. Characterization of the films by energy-dispersive X-ray spectroscopy and X-ray photoelectron spectroscopy confirmed excellent stoichiometric control of each of the 21 complex metal oxide films investigated. In studying the electrochemical oxidation of water catalyzed by the respective films, it was found that small concentrations of iron produced a significant improvement in Tafel slopes and that cobalt or nickel were critical in lowering the voltage at which catalysis commences. The best catalytic parameters of the series were obtained for the film of composition a-Fe20Ni80. An extrapolation of the electrochemical and XPS data indicates the optimal behavior of this binary film to be a manifestation of iron stabilizing nickel in a higher oxidation level. This work represents the first mechanistic study of amorphous phases of binary and ternary metal oxides for use as water oxidation catalysts, and provides the foundation for the broad exploration of other mixed-metal oxide combinations.


Effects of interleukins on EGF-stimulated growth promotion in human thyroid cells: differential modifications by IL-2 and IL-6 in Graves' and normal thyroid cells.
To investigate the role of interleukins on the growth of human thyroid cells, the effect of IL-2, IL-4 and IL-6 was studied on EGF-induced [3H]-thymidine uptake, cell cycle by flow cytometry, and mRNA levels of cellular proto-oncogene c-fos in thyroid monolayer cells from eighteen patients with Graves' disease and sixteen with non-thyroidal disease. EGF stimulated the DNA synthesis and the expression of c-fos mRNA both in Graves' thyroid cells and in normal thyroid cells. A dose-dependent inhibition of Graves' thyroid cell growth was observed with increasing concentration of IL-2 regardless of coculture with EGF. IL-2 did not influence [3H]-thymidine uptake nor the percentage of S+G2/M phase in cell cycle in normal thyroid cells. Such effects were not modified by the elimination or addition of lymphocytes. IL-2 did not affect the EGF-induced expression of c-fos mRNA in both Graves' and normal thyroid cells. IL-6 (10(-2)-10 ng/ml) stimulated the [3H]-thymidine uptake up to 218-303 % of control level, and the percentage of cells in S+G2/M phase was increased in IL-6-treated normal thyroid cells as compared to EGF-treated cells. IL-6 did not augment these parameters in Graves' thyroid cells. When EGF was included with IL-6, the level in c-fos mRNA was further augmented in normal thyroid cells. Such an additive effect was not seen in Graves' thyroid cells. Incubation of Graves' or normal thyroid cells with IL-4 did not affect the [3H]-thymidine uptake nor the percentage of S+G2/M. These data suggest that, compared with normal cells, the growth factors (such as cytokines), may act in an opposite way in Graves' thyroid cells. The different behavior between Graves' and normal thyroid cells in response to IL-2 and IL-6 might contribute to the pathogenesis of goiter formation.


4-Cyanamido-substituted benzenesulfonamides act as dual carbonic anhydrase and cathepsin inhibitors.
A set of novel N-cyano-N-substituted 4-aminobenzenesulfonamide derivatives were synthesized and investigated for their inhibitory activity against four cytosolic carbonic anhydrase (CA, EC 188.8.131.52) isoforms (hCA I, II, VII and XIII) and two cathepsins (S and B). N-alkyl/benzyl-substituted derivatives were revealed to be very potent inhibitors against brain-associated hCA VII, but inactive against both cathepsins. On the other hand, N-acyl-substituted derivatives displayed significant inhibitory activities against cathepsin S, but only moderate to poor inhibitory potency against hCA VII. Both hCA VII and cathepsin S have recently been validated as therapeutic targets in neuropathic pain. This study provided an excellent starting point for further structural optimization of this class of bifunctional compounds to enhance their inhibitory activity and selectivity against hCA VII and cathepsin S and to achieve new compounds with an attractive dual mechanism of action as anti-neuropathic agents.


Potential environmental impacts of light-emitting diodes (LEDs): metallic resources, toxicity, and hazardous waste classification.
Light-emitting diodes (LEDs) are advertised as environmentally friendly because they are energy efficient and mercury-free. This study aimed to determine if LEDs engender other forms of environmental and human health impacts, and to characterize variation across different LEDs based on color and intensity. The objectives are as follows: (i) to use standardized leachability tests to examine whether LEDs are to be categorized as hazardous waste under existing United States federal and California state regulations; and (ii) to use material life cycle impact and hazard assessment methods to evaluate resource depletion and toxicity potentials of LEDs based on their metallic constituents. According to federal standards, LEDs are not hazardous except for low-intensity red LEDs, which leached Pb at levels exceeding regulatory limits (186 mg/L; regulatory limit: 5). However, according to California regulations, excessive levels of copper (up to 3892 mg/kg; limit: 2500), Pb (up to 8103 mg/kg; limit: 1000), nickel (up to 4797 mg/kg; limit: 2000), or silver (up to 721 mg/kg; limit: 500) render all except low-intensity yellow LEDs hazardous. The environmental burden associated with resource depletion potentials derives primarily from gold and silver, whereas the burden from toxicity potentials is associated primarily with arsenic, copper, nickel, lead, iron, and silver. Establishing benchmark levels of these substances can help manufacturers implement design for environment through informed materials substitution, can motivate recyclers and waste management teams to recognize resource value and occupational hazards, and can inform policymakers who establish waste management policies for LEDs.


An improved treatment of empirical dispersion and a many-body energy decomposition scheme for the explicit polarization plus symmetry-adapted perturbation theory (XSAPT) method.
We recently introduced a low-cost quantum chemistry method for computing intermolecular interactions, combining a monomer-based self-consistent field calculation (the "explicit polarization" method, XPol) with pairwise-additive symmetry adapted perturbation theory (SAPT). The method uses Kohn-Sham (KS) orbitals in the SAPT formalism but replaces the SAPT dispersion and exchange-dispersion terms with empirical potentials ("+D"), and we called this method XPol+SAPT(KS)+D. Here, we report a second-generation version of this approach, XPol+SAPT(KS)+D2 or XSAPT(KS)+D2 for short, in which we have modified the form of the empirical atom-atom dispersion potentials. Accurate binding energies are obtained for benchmark databases of dimer binding energies, and potential energy curves are captured accurately for a variety of challenging systems. We suggest that using different asymptotic corrections for different monomers is necessary to get good binding energies in general, especially for hydrogen-bonded complexes. As compared to our original "+D" formulation, the second-generation "+D2" method accurately reproduces not only total binding energies but also the various components of the interaction energy, and on this basis we introduce an energy decomposition scheme that extends traditional SAPT energy decomposition to systems containing more than two monomers. For (H2O)6, the many-body contribution to the interaction energy agrees well with that obtained from traditional Kitaura-Morokuma energy decomposition analysis in a large basis set.


Synaptic vesicle capture by CaV2.2 calcium channels.
The fusion of synaptic vesicles (SVs) at the presynaptic transmitter release face is gated by Ca(2) (+) influx from nearby voltage-gated calcium channels (CaVs). Functional studies favor a direct molecular "tethering" attachment and recent studies have proposed a direct link to the channel C-terminal. To test for direct CaV-SV attachment we developed an in vitro assay, termed SV pull-down (SV-PD), to test for capture of purified, intact SVs. Antibody-immobilized presynaptic or expressed CaV2.2 channels but not plain beads, IgG or pre-blocked antibody successfully captured SVs, as assessed byWestern blot for a variety of protein markers. SV-PD was also observed with terminal fusion proteins of the distal half of the C-terminal, supporting involvement of this CaV region in tethering. Thus our results support a model in which the SV tethers directly to the CaV. Since the tip of the C-terminal could extend as far as 200 nm into the cytoplasm, we hypothesize that this link may serve as the initial SV capture mechanism by the release site. Further studies will be necessary to evaluate the molecular basis of C-terminal tethering and whether the SV binds to the channel by additional, shorter-range attachments.


Native T1 Mapping in the Diagnosis of Cardiac Allograft Rejection: A Prospective Histologically Validated Study.
This study aimed to determine the role of T1 mapping in identifying cardiac allograft rejection. Endomyocardial biopsy (EMBx), the current gold standard to diagnose cardiac allograft rejection, is associated with potentially serious complications. Cardiac magnetic resonance (CMR)-based T1 mapping detects interstitial edema and fibrosis, which are important markers of acute and chronic rejection. Therefore, T1 mapping can potentially diagnose cardiac allograft rejection noninvasively. Patients underwent CMR within 24 h of EMBx. T1 maps were acquired at 1.5-T. EMBx-determined rejection was graded according to International Society of Heart and Lung Transplant (ISHLT) criteria. Of 112 biopsies with simultaneous CMR, 60 were classified as group 0 (ISHLT grade 0), 35 as group 1 (ISHLT grade 1R), and 17 as group 2 (2R, 3R, clinically diagnosed rejection, antibody-mediated rejection). Native T1 values in patients with grade 0 biopsies and left ventricular ejection fraction >60% (983 ± 42 ms; 95% confidence interval: 972 to 994 ms) were comparable to values in nontransplant healthy control subjects (974 ± 45 ms; 95% confidence interval: 962 to 987 ms). T1 values were significantly higher in group 2 (1,066 ± 78 ms) versus group 0 (984 ± 42 ms; p = 0.0001) and versus group 1 (1,001 ± 54 ms; p = 0.001). After excluding patients with an estimated glomerular filtration rate <50 ml/min/m2, there was a moderate correlation of log-transformed native T1 with high-sensitivity troponin T (r = 0.54, p < 0.0001) and pro-B-type natriuretic peptide (r = 0.67, p < 0.0001). Using a T1 cutoff value of 1,029 ms, the sensitivity, specificity, and negative predictive value were 93%, 79%, and 99%, respectively. Myocardial tissue characterization with T1 mapping displays excellent negative predictive capacity for the noninvasive detection of cardiac allograft rejection and holds promise to reduce substantially the EMBx requirement in cardiac transplant rejection surveillance.


[Effect of pregnancy on metastasizing breast cancer--a case report].
The case of a 39 year old III gravid, I parous woman is reported. Four years ago she had had breast cancer (T2N1MO). Three years later she suffered from recurrence and a tumor was removed from the right axilla. One year later she became pregnant and presented with multiple bone metastases in her 5th month of pregnancy. During the following time an obvious progression could be shown. Therefore, a caesarean section with ovariectomy was performed at the beginning of 29th week of gestation. Postoperatively the patient had a partial remission. It is argued, that this could have been caused by the interaction of pregnancy and malignant disease, especially by the influence of growth-promoting estrogens.


In-vivo X-ray Dark-Field Chest Radiography of a Pig.
X-ray chest radiography is an inexpensive and broadly available tool for initial assessment of the lung in clinical routine, but typically lacks diagnostic sensitivity for detection of pulmonary diseases in their early stages. Recent X-ray dark-field (XDF) imaging studies on mice have shown significant improvements in imaging-based lung diagnostics. Especially in the case of early diagnosis of chronic obstructive pulmonary disease (COPD), XDF imaging clearly outperforms conventional radiography. However, a translation of this technique towards the investigation of larger mammals and finally humans has not yet been achieved. In this letter, we present the first in-vivo XDF full-field chest radiographs (32 × 35 cm2) of a living pig, acquired with clinically compatible parameters (40 s scan time, approx. 80 µSv dose). For imaging, we developed a novel high-energy XDF system that overcomes the limitations of currently established setups. Our XDF radiographs yield sufficiently high image quality to enable radiographic evaluation of the lungs. We consider this a milestone in the bench-to-bedside translation of XDF imaging and expect XDF imaging to become an invaluable tool in clinical practice, both as a general chest X-ray modality and as a dedicated tool for high-risk patients affected by smoking, industrial work and indoor cooking.


Genetic control of retinal specification and determination in Drosophila.
The Drosophila compound eye has long served as an outstanding model system to study many processes, including cell fate specification, cell division, cell growth and cell death. In addition, exploring the molecular basis of eye specification in Drosophila has identified a set of nuclear factors that trigger the conversion of a group of multipotent epithelial cells into eye primordia. These nuclear factors act in complex networks to regulate retinal specification and appear to be conserved throughout phylogeny. Finally, evidence suggests that these nuclear networks have been co-opted to specify cell fates in other tissues. We review the latest developments in the field of retinal specification in Drosophila and discuss several future directions that remain open for investigation.


Effects of beta-endorphin, vasoactive intestinal polypeptide and cholecystokinin octapeptide on cat carotid chemoreceptor activity.
The effects of beta-endorphin, vasoactive intestinal polypeptide (VIP) and cholecystokinin octapeptide (CCK-8) on carotid chemoreceptor activity have been investigated in cats anaesthetized with pentobarbitone. Spontaneous chemoreceptor discharge was decreased by intracarotid injection of beta-endorphin and by low doses of VIP, whereas it was increased by CCK-8 and higher doses of VIP, these effects being relatively long-lasting and often associated with changes in systemic blood pressure. The chemoexcitation evoked by acetylcholine and sodium cyanide was reduced during intracarotid infusion of any of the three peptides studied, and that caused by CO2-saturated Locke solution was reduced by beta-endorphin, largely unaltered by VIP and variably affected by CCK-8. The inhibitory effect of beta-endorphin was greatly reduced by naloxone, implying that it probably involved actions at naloxone-sensitive opiate receptors in the carotid body. Substance P was unable to overcome the chemoinhibitory effect of methionine enkephalin. Possible functions of polypeptides in the carotid body are discussed.


Enantiomeric separation of malathion and malaoxon and the chiral residue analysis in food and environmental matrix.
Malathion is a widely used chiral phosphorus insecticide, which has a more toxic chiral metabolite malaoxon. In this work, the enantiomers of malathion and malaoxon were separated by high-performance liquid chromatography-mass/mass (HPLC-MS/MS) with chiral columns using acetonitrile/water or methanol/water as mobile phase, and the chromatographic conditions were optimized. Based on the chiral separation, the chiral residue analysis methods for the enantiomers in soil, fruit, and vegetables were set up. Two pairs of the enantiomers were better separated on CHIRALPAK IC chiral column, and baseline simultaneous separations of malathion and malaoxon enantiomers were achieved with acetonitrile/water (40/60, v/v) as mobile phase at a flow rate of 0.5 mL/min. The elution orders were -/+ for both malathion and malaoxon measured by an optical rotation detector. The chiral residue analysis in soil, fruit, and vegetables was validated by linearity, recovery, precision, limit of detection (LOD), and limit of quantification (LOQ). The LODs and LOQs for the enantiomers of malathion were 1 μg/kg and 3-5 μg/kg and 0.08 μg/kg and 0.20-0.25 μg/kg for malaoxon enantiomers. Good linear calibration curves for each enantiomer in the matrices were obtained within the concentration range of 0.02-12 mg/L. The mean recoveries of the enantiomers of malathion and malaoxon ranged from 82.26% to 109.04%, with RSDs of 0.71-8.63%.The results confirmed that this method was capable of simultaneously determining the residue of malathion and malaoxon in food and environmental matrix on an enantiomeric level.


Sirenomelia: a case report.
Sirenomelia is a rare and interesting congenital anomaly where there is fusion of the lower limb buds in association with a number of other anomalies, together termed as the caudal regression syndrome. We report this syndrome in a male foetus of 33 weeks gestational age, and review its embryological basis.


[Shortening SRT of Intermittent Gradient Aeration to Realize Nitrogen and Phosphorus Removal in Short-range SNEDPR System].
This study explored the effect of sludge retention time (SRT) on ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) under intermittent gradient aeration, as well as the effect of the short-range nitrification endogenous denitrification phosphorus removal system on the treatment of low C/N ratio domestic sewage. In this study, an SBR reactor was used to cultivate aerobic granular sludge, using actual domestic sewage as the influent substrate. As the SRT decreased from 50 d to 30 d, the specific ammonia oxidation rate increased from 3.16 mg·(g·h)-1to 4.38 mg·(g·h)-1, and the specific nitrite oxidation rate decreased from 3.4 mg·(g·h)-1to 1.8 mg·(g·h)-1. The activity of NOB decreased by about 44%, resulting in short-range nitrification within the system. With an SRT of 30 d, the maximum nitrite accumulation was 6.93 mg·L-1. Because the reduced SRT led to a slight decrease in sludge concentration within the system, an aeration reduction strategy was adopted after 40 d, according to the DO curve. When the final SRT was 30 d, the effluent COD concentration was 40.76 mg·L-1, the TN concentration was 12.4 mg·L-1, the TP concentration was 0.31 mg·L-1, and the simultaneous removal of C, N and P was realized. Thus, a stable short-range nitrification endogenous denitrification phosphorus removal system was finally obtained. At the same time, the EPS content of aerobic granular sludge was negatively correlated with SRT, the protein content increased from 66.7 mg·g-1 to 95.1 mg·g-1, and the polysaccharide content remained in the range of 12.1-17.2 mg·g-1, indicating that the decreased SRT had a great effect on the protein content. With an SRT of 30 d, the PN/PS value was maintained at approximately 6.2, and the structural stability of aerobic granular sludge can be maintained under such conditions.


Electric-Field Assisted In Situ Hydrolysis of Bulk Metal-Organic Frameworks (MOFs) into Ultrathin Metal Oxyhydroxide Nanosheets for Efficient Oxygen Evolution.
Facile preparation of low-cost electrocatalysts for efficient oxygen evolution reaction (OER) remains a big challenge. Herein, a novel strategy for ultrafast (20 s) transformation of bulk metal-organic frameworks (MOFs) into ultrathin metal oxyhydroxide nanosheets for efficient OER has been developed. For two isomeric MOFs (FJI-H25Fe and FJI-H25FeCo), only the metastable FJI-H25FeCo bulk can immediately transform into FeCo-oxyhydroxides nanosheets through electric-field assisted hydrolysis. The potential evolution process from MOF bulk to FeCo-oxyhydroxides nanosheets has been investigated in detail. The as-made nanosheets exhibit excellent OER performances, showing an extremely low overpotential of 231 mV at the current density of 10 mA cm-2 , a relatively small Tafel slope of 42 mV dec-1 , and long-term durability of at least 30 h. This work not only provides a novel strategy for facile preparation of low-cost and efficient OER electrocatalysts, but also represents a new way for preparation of metal oxyhydroxides nanosheets with good crystallinity and morphology, and a fresh method for mild synthesis of nanosized derivatives from MOF materials.


Predictors of neurobehavioral symptoms in a university population: a multivariate approach using a postconcussive symptom questionnaire.
Several factors have been linked to severity of postconcussive-type (neurobehavioral) symptoms. In this study, predictors of neurobehavioral symptoms were examined using multivariate methods to determine the relative importance of each. Data regarding demographics, symptoms, current alcohol use, history of traumatic brain injury (TBI), orthopedic injuries, and psychiatric/developmental diagnoses were collected via questionnaire from 3027 university students. The most prominent predictors of symptoms were gender, history of depression or anxiety, history of attention-deficit/hyperactivity disorder or learning disability diagnosis, and frequency of alcohol use. Prior mild TBI was significantly related to overall symptoms, but this effect was small in comparison to other predictors. These results provide further evidence that neurobehavioral symptoms are multi-determined phenomena, and highlight the importance of psychiatric comorbidity, demographic factors, and health behaviors to neurobehavioral symptom presentation after mild TBI.


The Effect of Skin-to-Skin Contact on Placental Separation Time and Initiation of Breastfeeding.
Background: The first hour and minutes immediately after birth are quite important for both the mother and the newborn. Ensuring skin-to-skin contact (SSC) during this period has a positive effect on the mother both psychologically and physiologically, and it is also important in starting breastfeeding. Objective: The study was conducted to examine the effect of SSC on placental separation time and initiation of breastfeeding. Materials and Methods: This is a randomized controlled study. The sample of the study consists of a total of 84 women with primiparous birth and vaginal delivery, including 42 experimental and 42 controls. Data were collected using a personal information form, a chronometer, and the LATCH scale. The placental separation time for those in the experimental and control groups was determined by using a chronometer. Initiation and assessment of breastfeeding were evaluated at the first and 24th hours of birth using the LATCH scale. Results: The signs of placental separation occurred earlier in the experimental group than in the control group. The mean placental separation time was found to be 7.05 ± 4.37 minutes in the experimental group and 13.41 ± 5.24 minutes in the control group. A statistically significant difference was found between the groups in terms of placental separation time (p = 0.0001). Conclusion: This study concludes that SSC in the third stage of labor ensures earlier separation of the placenta and is effective in initiating breastfeeding.


Dynamic use of multiple parameter sets in sequence alignment.
The level of conservation between two homologous sequences often varies among sequence regions; functionally important domains are more conserved than the remaining regions. Thus, multiple parameter sets should be used in alignment of homologous sequences with a stringent parameter set for highly conserved regions and a moderate parameter set for weakly conserved regions. We describe an alignment algorithm to allow dynamic use of multiple parameter sets with different levels of stringency in computation of an optimal alignment of two sequences. The algorithm dynamically considers various candidate alignments, partitions each candidate alignment into sections, and determines the most appropriate set of parameter values for each section of the alignment. The algorithm and its local alignment version are implemented in a computer program named GAP4. The local alignment algorithm in GAP4, that in its predecessor GAP3, and an ordinary local alignment program SIM were evaluated on 257,716 pairs of homologous sequences from 100 protein families. On 168,475 of the 257,716 pairs (a rate of 65.4%), alignments from GAP4 were more statistically significant than alignments from GAP3 and SIM.


Dissociable Fronto-Operculum-Insula Control Signals for Anticipation and Detection of Inhibitory Sensory Cue.
The ability to anticipate and detect behaviorally salient stimuli is important for virtually all adaptive behaviors, including inhibitory control that requires the withholding of prepotent responses when instructed by external cues. Although right fronto-operculum-insula (FOI), encompassing the anterior insular cortex (rAI) and inferior frontal cortex (rIFC), involvement in inhibitory control is well established, little is known about signaling mechanisms underlying their differential roles in detection and anticipation of salient inhibitory cues. Here we use 2 independent functional magnetic resonance imaging data sets to investigate dynamic causal interactions of the rAI and rIFC, with sensory cortex during detection and anticipation of inhibitory cues. Across 2 different experiments involving auditory and visual inhibitory cues, we demonstrate that primary sensory cortex has a stronger causal influence on rAI than on rIFC, suggesting a greater role for the rAI in detection of salient inhibitory cues. Crucially, a Bayesian prediction model of subjective trial-by-trial changes in inhibitory cue anticipation revealed that the strength of causal influences from rIFC to rAI increased significantly on trials in which participants had higher anticipation of inhibitory cues. Together, these results demonstrate the dissociable bottom-up and top-down roles of distinct FOI regions in detection and anticipation of behaviorally salient cues across multiple sensory modalities.


Tacrolimus ameliorates cerebral vasodilatation and intracranial hypertension in the rat with portacaval anastomosis and hyperammonemia.
Arterial hyperammonemia and cerebral vasodilatation correlate with cerebral herniation in patients with fulminant hepatic failure (FHF). Tacrolimus is a calcineurin inhibitor that passes the blood-brain barrier and may increase cerebrovascular tone and restrict cerebral ammonia influx. In this study, we determined if tacrolimus prevents cerebral vasodilatation and high intracranial pressure (ICP) in the rat with portacaval anastomosis (PCA) challenged to high arterial ammonia (NH4+) concentration. Seven groups of mechanically ventilated rats, with 6-9 rats in each group, were investigated within 48 hours after construction of a PCA (4 groups) or after sham operation (3 groups). Three groups of the rats received infusion of NH4+ and 4 groups received saline for approximately 180 minutes. Two groups of the PCA rats receiving either NH4+ or saline had an i.v. injection of tacrolimus (0.4 mg/kg) or vehicle before start of NH4+ or saline infusion. Cerebral blood flow (CBF) was monitored by a laser Doppler probe in brain cortex. ICP was monitored by placement of a catheter in the cerebrospinal fluid. CBF and ICP increased in PCA rats receiving NH4+ infusion compared to PCA controls and to all groups of sham-operated animals (P <.05). In the group of PCA rats pre-treated with tacrolimus before receiving ammonia infusion, the increase in ICP was ameliorated compared to the ammonia infused group receiving vehicle (P <.03). Tacrolimus also prevented an increase in CBF in the PCA group receiving NH4+ (P <.05) compared to the control groups. In conclusion, Tacrolimus prevents cerebral vasodilatation and ameliorates intracranial hypertension in PCA rats receiving NH4+ infusion. These findings indicate that tacrolimus could be of clinical value in the prevention of cerebral hyperemia, high ICP, and serious brain damage in patients with FHF.


Prevalence of left-handedness among patients with different respiratory diseases.
We investigated the prevalence of left-handedness among patients with different respiratory diseases. Hand preference was assessed using the modified Edinburg Handedness Inventory. There was a statistically significant decline in left-handedness with shortened life span in patients. Difference among the rates of left-handedness in different pulmonary diseases was significant. The rate of left-handedness was lowest in patients with chronic obstructive pulmonary disease (2.14%) and pneumonia (2.04%) and highest in patients with asthma (12.96%). The results suggest that left-handed subjects may be protected from certain pulmonary diseases while being more susceptible to others.


Coping and social support in asthma and allergy: the Västerbotten Environmental Health Study.
Asthma and allergy are stressful conditions that require coping strategies and social support to reduce stress and enhance health-promoting behavior. However, research is limited regarding coping and social support in asthma and allergy. The aim was to better understand the use of different coping strategies and perceived social support in low and high severity (exacerbation frequency) of asthma and allergy. Population-based data were used to provide ratings of coping strategies (Study I) and social support (Study II) from 124 and 94 participants, respectively, with asthma and/or allergy, categorized as low or high in severity. Problem- and emotion-focused coping strategies were assessed as well as emotional, instrumental and informative social support from seven sources. Study I showed that avoiding certain environments (problem-based coping) and trying to accept one's situation (emotion-based) were the most commonly used coping strategies. These behaviors did not differ due to severity. Study II showed that more emotional than instrumental and informative support was perceived. The highest rated support sources were the partner, family members and the healthcare system. More social support was reported in low asthma/allergy severity compared to high asthma/allergy severity. The most commonly used coping strategies in the population of persons with these four types of asthma and allergy are avoiding certain environments and trying to accept one's situation. More emotional support than instrumental and informative is perceived to be received, and most of the support is received from one's partner and other family members and least from authorities and patient associations/support groups.


A novel mutation (E333D) in the thyroid hormone beta receptor causing resistance to thyroid hormone syndrome.
Resistance to thyroid hormone (RTH) is an inherited syndrome characterized by elevated serum thyroid hormones (TH), failure to suppress pituitary thyroid stimulating hormone (TSH) secretion, and variable peripheral tissue responsiveness to TH. The disorder is associated with diverse mutations in the thyroid hormone beta receptor (TRbeta). Here, we report a novel natural RTH mutation (E333D) located in the large carboxy-terminal ligand binding domain of TRbeta. The mutation was identified in a 22-year-old French woman coming to medical attention because of an increasing overweight. Biochemical tests showed elevated free thyroxine (T4: 20.8 pg/ml (normal, 8.5-18)) and triiodothyronine (T3: 5.7 pg/ml (normal, 1.4-4)) in the serum, together with an inappropriately nonsuppressed TSH level of 4.7 mU/ml (normal, 0.4-4). Her father and her brother's serum tests also showed biochemical abnormalities consistent with RTH. Direct sequencing of the TRbeta gene revealed a heterozygous transition 1284A>C in exon 9 resulting in substitution of glutamic acid 333 by aspartic acid residue (E333D). Further functional analyses of the novel TRbeta mutant were conducted. We found that the E333D mutation neither significantly affected the affinity of the receptor for T3 nor modified heterodimer formation with retinoid X receptor (RXR) when bound to DNA. However, in transient transfection assays, the E333D TRbeta mutant exhibited impaired transcriptional regulation on two distinct positively regulated thyroid response elements (F2- and DR4-TREs) as well as on the negatively regulated human TSHalpha promoter. Moreover, a dominant inhibition of the wild-type TRbeta counterpart transactivation function was observed on both a positive (F2-TRE) and a negative (TSHalpha) promoter. These results strongly suggest that the E333D TRbeta mutation is responsible for the RTH phenotype in the proposita's family.


Non-invasive prenatal paternity testing by analysis of Y-chromosome mini-STR haplotype using next-generation sequencing.
To assess the efficacy of Y-chromosome mini-STR-based next-generation sequencing (NGS) for non-invasive prenatal paternity testing (NIPPT). DNA was extracted from the plasma of 24 pregnant women, and cell-free fetal DNA (cffDNA) haplotyping was performed at 12 Y-chromosome mini-STR loci using the Illumina NextSeq 500 system. The cffDNA haplotype was validated by the paternal haplotype. Subsequentlly, the paternity testing parameters were attributed to each case quantitatively. The biological relationship between the alleged fathers and infants in all 24 family cases were confirmed by capillary electrophoresis (CE). The Y-chromosome mini-STR haplotypes of all 14 male cffDNA were obtained by NGS without any missing loci. The alleles of cffDNA and paternal genomic DNA were matched in 13 cases, and a mismatched allele was detected at the DYS393 locus in one case and considered as mutation. No allele was detected in the 10 female cffDNA. The combined paternity index (CPI) and probability of paternity calculation was based on 6 loci Y-haplotype distributions of a local population. The probability of paternity was 98.2699-99.8828% for the cases without mutation, and 14.8719% for the case harboring mutation. Our proof-of-concept study demonstrated that Y-chromosome mini-STR can be used for NGS-based NIPPT with high accuracy in real cases, and is a promising tool for familial searching, paternity exclusion and sex selection in forensic and medical applications.


Task-related modulation of effective connectivity during perceptual decision making: dissociation between dorsal and ventral prefrontal cortex.
The dorsal and ventral parts of the lateral prefrontal cortex have been thought to play distinct roles in decision making. Although its dorsal part such as the frontal eye field (FEF) is shown to play roles in accumulation of sensory information during perceptual decision making, the role of the ventral prefrontal cortex (PFv) is not well-documented. Previous studies have suggested that the PFv is involved in selective attention to the task-relevant information and is associated with accuracy of the behavioral performance. It is unknown, however, whether the accumulation and selection processes are anatomically dissociated between the FEF and PFv. Here we show that, by using concurrent TMS and EEG recording, the short-latency (20-40 ms) TMS-evoked potentials after stimulation of the FEF change as a function of the time to behavioral response, whereas those after stimulation of the PFv change depending on whether the response is correct or not. The potentials after stimulation of either region did not show significant interaction between time to response and performance accuracy, suggesting dissociation between the processes subserved by the FEF and PFv networks. The results are consistent with the idea that the network involving the FEF plays a role in information accumulation, whereas the network involving the PFv plays a role in selecting task relevant information. In addition, stimulation of the FEF and PFv induced activation in common regions in the dorsolateral and medial frontal cortices, suggesting convergence of information processed in the two regions. Taken together, the results suggest dissociation between the FEF and PFv networks for their computational roles in perceptual decision making. The study also highlights the advantage of TMS-EEG technique in investigating the computational processes subserved by the neural network in the human brain with a high temporal resolution.


Borrelidin Isolated from Streptomyces sp. Inhibited Adipocyte Differentiation in 3T3-L1 Cells via Several Factors Including GATA-Binding Protein 3.
An inhibitor of 3T3-L1 adipocyte differentiation was isolated from Streptomyces sp. TK08330 and identified by spectroscopy as the 18-membered macrolide borrelidin. Treatment with 1.0 μM borrelidin suppressed intracellular lipid accumulation by 80% and inhibited the expression of adipocyte-specific genes. Borrelidin suppressed the mRNA expression of two master regulators of adipocyte differentiation, peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer binding protein (C/EBPα). Studies on well-known upstream regulators of PPARγ revealed that borrelidin down-regulated C/EBPδ mRNA expression but did not affect expression of C/EBPβ. Borrelidin increased mRNA expression of negative regulators of differentiation such as GATA-binding protein (GATA) 3, Krüppel-like factor (KLF) 3 and KLF7, as well as positive regulators, KLF4, KLF6 and KLF15, at early stages of differentiation. To elucidate a primary mediator of borrelidin differentiation inhibitory activity, small interfering RNA (siRNA) transfection experiments were performed. The mRNA expression of PPARγ, which was down-regulated by borrelidin, was not changed by KLF3 and KLF7 siRNA treatment. In contrast, expression of PPARγ in GATA-3 siRNA-treated cells was not significantly different from that of control siRNA-treated cells. Borrelidin significantly inhibited lipid accumulation in control siRNA-treated cells, and treatment with GATA-3 siRNA slightly reduced the inhibitory effect of borrelidin. These results indicate that borrelidin inhibited adipocyte differentiation partially via GATA-3.


Lessons to be learned: a case study approach--acute appendicitis masquerading as macroamylasaemia.
Macroamylasaemia is a condition in which serum amylase is elevated in the presence of a low to normal urinary amylase and normal renal function. It is rare but can masquerade as other clinical disorders. Discussed here is a case report of a patient who presented initially with abdominal pain (later recognized as being due to gangrenous appendicitis) and in whom there was a very high serum amylase level, leading to an erroneous initial diagnosis and management as acute pancreatitis. The CT scan of the abdomen was normal without any evidence of pancreatitis. Subsequently, the renal amylase:creatinine clearance ratio (C(am)/C(cr)) was found to be low, being characteristic and diagnostic of macroamylasaemia; the latter was, in turn, the cause for the elevated serum amylase level. The underlying macroamylasaemia had thus masqueraded as pancreatitis. The patient underwent appendicectomy and hence made an excellent recovery. It is vitally important to recognize this condition in order to avoid both an incorrect diagnosis and inappropriate treatment/management.


'Lost': listening to the voices and mental health needs of forced migrants in London.
Research into the mental health needs of asylum seekers and refugees has revealed that they are likely to experience poorer mental health as well as higher levels of exclusion and vulnerability than native populations. This paper reports on data drawn from semi-structured interviews of 21 refugees and asylum seekers that describe the complexity experienced by those living in exile, and the necessity for a more integrated and holistic approach in the planning and delivery of services to support mental health. Incorporating a perspective from service users will encourage providers to take account of the multitude of practical, social, cultural, economic and legal difficulties that can influence the long-term mental health of this population. The implications highlight a need to shift from a simple biomedical model of the causes and effects of ill-health to a social model, which will require reorganisation not only in healthcare but in welfare, housing, employment and immigration policy.


In vitro studies with cefazaflur and other parenteral cephalosporins.
Cefazaflur has a broad-spectrum of in vitro antibacterial activity equal to or greater than that of the commercially-available cephalosporins. In addition, cefazaflur has activity against isolates of Enterobacter, Citrobacter and indole-positive Proteus; however, this activity decreased markedly when the MIC determinations were carried out with a large inoculum size. A similar inoculum effect was observed with cefamandole, however, cefoxitin's activity was relatively unchanged at increased inoculum sizes. Human serum had a relatively small effect on the in vitro activity of cefazaflur.


The effect of induced optimism on early pain processing: indication by contact heat evoked potentials (CHEPs) and the sympathetic skin response (SSR).
Situationally induced optimism has been shown to influence several components of experimental pain. The aim of the present study was to enlarge these findings for the first time to the earliest components of the pain response by measuring contact heat evoked potentials (CHEPs) and the sympathetic skin response (SSR). Forty-seven healthy participants underwent two blocks of phasic thermal stimulation. CHEPs, the SSR and self-report pain ratings were recorded. Between the blocks of stimulation, the 'Best Possible Self' imagery and writing task was performed to induce situational optimism. The optimism manipulation was successful in increasing state optimism. It did, however, neither affect pain-evoked potentials nor the SSR nor self-report pain ratings. These results suggest that optimism does not alter early responses to pain. The higher-level cognitive processes involved in optimistic thinking might only act on later stages of pain processing. Therefore, more research is needed targeting different time frames of stimulus processing and response measures for early and late pain processing in parallel.


Hypnotic underestimation of time: the busy beaver hypothesis.
Two experiments tested the hypothesis that the hypnotic underestimation of time is mediated by attentional processing. In Experiment 1, variations in the demands placed on attentional resources produced substantial differences in the subjective estimates of identical length intervals occurring within a hypnotic context. In Experiment 2, attentional manipulation was assessed in both hypnotic and waking contexts. Time judgments were again found to vary with attentional demands but not with hypnotic context. The results are consistent with a busy beaver hypothesis, which holds that hypnotic, as well as nonhypnotic, time estimates are a by-product of the attentional processing demands of the task.


Gene discovery using next-generation pyrosequencing to develop ESTs for Phalaenopsis orchids.
Orchids are one of the most diversified angiosperms, but few genomic resources are available for these non-model plants. In addition to the ecological significance, Phalaenopsis has been considered as an economically important floriculture industry worldwide. We aimed to use massively parallel 454 pyrosequencing for a global characterization of the Phalaenopsis transcriptome. To maximize sequence diversity, we pooled RNA from 10 samples of different tissues, various developmental stages, and biotic- or abiotic-stressed plants. We obtained 206,960 expressed sequence tags (ESTs) with an average read length of 228 bp. These reads were assembled into 8,233 contigs and 34,630 singletons. The unigenes were searched against the NCBI non-redundant (NR) protein database. Based on sequence similarity with known proteins, these analyses identified 22,234 different genes (E-value cutoff, e-7). Assembled sequences were annotated with Gene Ontology, Gene Family and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Among these annotations, over 780 unigenes encoding putative transcription factors were identified. Pyrosequencing was effective in identifying a large set of unigenes from Phalaenopsis. The informative EST dataset we developed constitutes a much-needed resource for discovery of genes involved in various biological processes in Phalaenopsis and other orchid species. These transcribed sequences will narrow the gap between study of model organisms with many genomic resources and species that are important for ecological and evolutionary studies.


The antihyperalgesic effects of intrathecal bupropion, a dopamine and noradrenaline reuptake inhibitor, in a rat model of neuropathic pain.
Antidepressants are often used for the treatment of neuropathic pain, and their analgesic effects rely on increased noradrenaline and serotonin levels in the spinal cord. Clinical studies have also shown that bupropion, a dopamine and noradrenaline reuptake inhibitor, has strong efficacy in neuropathic pain; however, the role of spinal cord dopamine in neuropathic pain is unknown. We hypothesized that bupropion inhibits neuropathic pain by increasing noradrenaline and dopamine in the spinal cord. In the present study, we determined the efficacy and underlying mechanisms of intrathecal administration of bupropion in a rat model of neuropathic pain. Male Sprague-Dawley rats were anesthetized, and right L5 spinal nerve ligation (SNL) was performed to produce mechanical hyperalgesia of the hindpaw. Withdrawal threshold to a paw pressure test was measured before and after intrathecal administration of bupropion, without or with intrathecal antagonists for α2-adrenoceptors and dopamine D2 receptors. In vivo microdialysis was performed in the dorsal horn of the lumbar spinal cord to measure noradrenaline and dopamine concentrations after intrathecal injection of bupropion. We also measured the noradrenaline and dopamine contents in the ipsilateral dorsal lumbar spinal cord in normal rats and in rats 2, 3, and 4 weeks after SNL. Intrathecal injection of bupropion produced a dose-dependent antihyperalgesic effect (3, 10, 30, and 100 μg, P < 0.001). The effect (30 μg) was dose-dependently reversed by intrathecal pretreatment (15 minutes before bupropion injection) with the α2-adrenoceptor antagonist idazoxan (3, 10, and 30 μg, P < 0.001) and D2 receptor antagonist sulpiride (3, 10, and 30 μg, P < 0.001). Microdialysis revealed that noradrenaline and dopamine concentrations in the spinal dorsal horn were increased after intrathecal injection of bupropion (30 μg, P < 0.001 and P = 0.001, respectively). Furthermore, the noradrenaline and dopamine contents in the spinal dorsal horn were increased 2 weeks after SNL (P < 0.001 and P = 0.044, respectively) and then decreased gradually. These findings suggest that plasticity of descending inhibitory pathways such as the noradrenaline and dopamine systems contributes to the maintenance of neuropathic pain and that spinal cord noradrenaline and dopamine both play an inhibitory role in neuropathic pain.


The Application of Nanomaterials in Stem Cell Therapy for Some Neurological Diseases.
Stem cell therapy provides great promising therapeutic benefits for various neurological disorders. Cell transplantation has emerged as cell replacement application for nerve damage. Recently, nanomaterials obtain wide development in various industrial and medical fields, and nanoparticles have been applied in the neurological field for tracking and treating nervous system diseases. Combining stem cells with nanotechnology has raised more and more attentions; and it has demonstrated that the combination has huge effects on clinical diagnosis and therapeutics in multiple central nervous system diseases, meanwhile, improves prognosis. The aim of this review was to give a brief overview of the application of nanomaterials in stem cell therapy for neurological diseases. Nanoparticles not only promote stem cell proliferation and differentiation in vitro or in vivo, but also play dominant roles on stem cell imaging and tracking. Furthermore, via delivering genes or drugs, nanoparticles can participate in stem cell therapeutic applications for various neurological diseases, such as ischemic stroke, spinal cord injury (SCI), multiple sclerosis (MS), Parkinson's disease (PD), Alzheimer's disease (AD) and gliomas. However, nanoparticles have potential cytotoxic effects on nerve cells, which are related to their physicochemical properties. Nano-stem cell-based therapy as a promising strategy has the ability to affect neuronal repair and regeneration in the central nervous system.


[Chikungunya outbreak on Reunion Island in 2005/2006: role of hospital physicians in raising alert].
The epidemic of chikungunya (CHIK) that swept through Reunion Island from late 2005 to mid 2006 affected 38.2% of the population, i.e., 300000 people. Although this outbreak took place in a French overseas department with high public health standards, failure to anticipate a large-scale epidemic associated with unprecedented severity and unexpectedly high mortality led to a major public health crisis. The purpose of this report is to provide a complete account of the experience of hospital intensive care physicians in addressing problems ranging from discovery of severe forms to management of a major health crisis. This report underlines the role of the head hospital physician and the necessity of mutual trust and collaboration with supervisory authorities.


Diastereoselective Synthesis of 1,2-Dihydrobenzofuro[3,2-b]pyridines via a Carbon-Carbon Double-Bond Cleavage/Rearrangement Cascade.
A new Lewis acid-catalyzed [2 + 2] cycloaddition/retroelectrocyclization (CA-RE)/1,6-addition relay of aurone-derived 1-azadienes and 1-alkynylnaphthalen-2-ols has been reported, leading to the regio- and diastereoselective synthesis of 1,2-dihydrobenzofuro[3,2-b]pyridine with a chiral carbon center and an axial chirality in good yields. This protocol enables the C-C double-bond scission/recombination to rapidly construct aza-heterocyclic architectures and features 100% atom utilization, a wide substrate scope, good compatibility with substituents, and excellent diastereoselectivity.


[Transfer and identification of multidrug resistance gene in mesenchymal stem cells derived from human placenta].
To explore the feasibility and safety of mdr1 gene transferred into placenta derived mesenchymal stem cells (P-MSCs) by reconstructed retroviral vector. Human P-MSCs were isolated and expanded by Percoll density gradient and then transduced repeatedly by reconstructed retroviral vector containing mdr1 gene. The transfection and expression of mdr1 gene was detected by reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS). Meanwhile, the biological features of mdr1-MSCs were identified and analyzed. The expression of mdr1mRNA was found in transfected cells. The expression of P-glycoprotein (P-gp) encoded by mdr1 gene was (27.6 ± 5.1)% in the transfected P-MSCs cells versus (0.4 ± 0.1)% in the non-transfected P-MSCs cells (t = 14.291, P < 0.01). The percent of P-MSCs at quiescent phase (G0/G1 phase) was around 95.40% and it was in accord with the characterization of stem cells. The mdr1-MSCs exhibited typical ultrastructures of low-differentiated stem cells. Moreover, they still retained the potency of adipogenic and osteogenic differentiation in the presence of appropriate conditioned media. A stable expression of P-gp may be obtained by reconstructed retroviral-mediated transfection in vitro. And transfected MSCs retain the characteristics of stem cells.


Smoothing spline-based score tests for proportional hazards models.
We propose "score-type" tests for the proportional hazards assumption and for covariate effects in the Cox model using the natural smoothing spline representation of the corresponding nonparametric functions of time or covariate. The tests are based on the penalized partial likelihood and are derived by viewing the inverse of the smoothing parameter as a variance component and testing an equivalent null hypothesis that the variance component is zero. We show that the tests have a size close to the nominal level and good power against general alternatives, and we apply them to data from a cancer clinical trial.


Interplay between RAGE and TLR4 Regulates HMGB1-Induced Inflammation by Promoting Cell Surface Expression of RAGE and TLR4.
Receptor for advanced glycation end-products (RAGE) and TLR4 play an important role in the inflammatory response against High-mobility group box 1 protein (HMGB1), a late proinflammatory cytokine and a damage-associated molecular pattern. As cell surface receptors, both RAGE and TLR4 are constantly trafficking between the cytoplasm and plasma membrane. However, whether TLR4 is related to the intracellular transport of RAGE in HMGB1-induced inflammation remains unknown. In this study, we demonstrated that HMGB1 not only increased RAGE expression in both the cytoplasm and plasma membrane but also upregulated the expression of TLR4 in the plasma membrane. Knocking out of RAGE led to decreased MAPK activation, TLR4 cellular membrane expression, and corresponding inflammatory cytokine generation. Meanwhile, inhibiting MAPK activation also decreased TLR4 surface expression. These results indicated that HMGB1 may bind to cell surface RAGE receptors on the cell surface, leading to MAPK activation, thus promoting TLR4 translocation on the cell surface, but does not regulate its transcription and translation. In contrast, TLR4 can increase the transcription and translation of RAGE, which translocates to the cell surface and is able to bind to more HMGB1. The cell surface receptors TLR4 and RAGE bind to HMGB1, leading to the transcription and secretion of inflammatory cytokines. Finally, we also observed these results in the mice pseudofracture model, which is closely related to HMGB1-induced inflammatory response. All these results demonstrated that the interplay between RAGE and TLR4 are critical for HMGB1-induced inflammatory response.


Biomimetic synthesis of 2D ultra-small copper sulfide nanoflakes based on reconfiguration of the keratin secondary structure for cancer theranostics in the NIR-II region.
Two-dimensional transition metal dichalcogenides have attracted widespread attention in cancer theranostics due to their high specific surface area and excellent photothermal conversion properties. However, their dimensions and biodegradability have limited the exploration of the therapeutic properties of transition metal dichalcogenides. Herein, we explore the mechanism of the keratin α-helix-to-random coil transition, as an actuation mechanism for the controllable design and precise synthesis of two-dimension copper sulfide nanoflakes (CuS NFs) with high absorption in the NIR-II window. Upon mixing keratin and Cu2+, the hydrogen bonds that maintain the α-helix are broken by copper ions to form biuret coordination, while the structure of the α-helix is transformed into a random coil, providing a more scalable space for the growth of CuS NFs. The CuS NFs prepared in this way possess the great advantages of outstanding uniformity, size controllability, and biodegradability. Importantly, the CuS NFs in the NIR-II window show an excellent photothermal conversion efficiency (32.9%) and extraordinary photoacoustic signal. This work updates the fabrication of two-dimensional transition metal dichalcogenides and greatly enhances their competitiveness in the area of cancer theranostics in the NIR-II region, and provides significant theoretical and practical opportunities for the development of keratin using biomimetic synthesis.


The extracellular phage-host interactions involved in the bacteriophage LL-H infection of Lactobacillus delbrueckii ssp. lactis ATCC 15808.
The complete genome sequence of Lactobacillus bacteriophage LL-H was determined in 1996. Accordingly, LL-H has been used as a model phage for the infection of dairy Lactobacillus, specifically for thermophilic Lactobacillus delbrueckii ssp. lactis host strains, such as ATCC 15808. One of the major goals of phage LL-H research consisted of the characterization of the first phage-host interactions at the level of phage adsorption and phage DNA injection steps to determine effective and practical methods to minimize the risks associated with the appearance and attack of phages in the manufacture of yogurt, and Swiss or Italian hard type cheeses, which typically use thermophilic lactic acid bacteria starter cultures containing L. delbrueckii strains among others. This mini review article summarizes the present data concerning (i) the special features, particle structure, and components of phage LL-H and (ii) the structure and properties of lipoteichoic acids (LTAs), which are the phage LL-H receptor components of L. delbrueckii ssp. lactis host strains. Moreover, a model of the first, extracellular, phage-host interactions for the infection of L. delbrueckii ssp. lactis ATCC 15808 by phage LL-H is presented and further discussed.


Mechanical strain determines the axis of planar polarity in ciliated epithelia.
Epithelia containing multiciliated cells align beating cilia along a common planar axis specified by the conserved planar cell polarity (PCP) pathway. Specification of the planar axis is also thought to require a long-range cue to align the axis globally, but the nature of this cue in ciliated and other epithelia remains poorly understood. We examined this issue using the Xenopus larval skin, where ciliary flow aligns to the anterior-posterior (A-P) axis. We first show that a planar axis initially arises in the developing skin during gastrulation, based on the appearance of polarized apical microtubules and cell junctions with increased levels of stable PCP components. This axis also arises in severely ventralized embryos, despite their deficient embryonic patterning. Because ventralized embryos still gastrulate, producing a mechanical force that strains the developing skin along the A-P axis, we asked whether this strain alone drives global planar patterning. Isolated skin explanted before gastrulation lacks strain and fails to acquire a global planar axis but responds to exogenous strain by undergoing cell elongation, forming polarized apical microtubules, and aligning stable components of the PCP pathway orthogonal to the axis of strain. The planar axis in embryos can be redirected by applying exogenous strain during a critical period around gastrulation. Finally, we provide evidence that apical microtubules and the PCP pathway interact to align the planar axis. These results indicate that oriented tissue strain generated by the gastrulating mesoderm plays a major role in determining the global axis of planar polarity of the developing skin.


[Influence of psychologic attitude to efficiency of pain treatment in vibration disease].
The article presents results of study concerning influence of desadaptive psychologic attitudes on formation and perception of pain syndrome in 148 vibration disease patients. The authors determined clinical and psychologic predictors of efficiency of pain syndrome treatment in vibration disease patients.


The poly-cistronic expression of four transcriptional factors (CRX, RAX, NEURO-D, OTX2) in fibroblasts via retro- or lentivirus causes partial reprogramming into photoreceptor cells.
The introduction of four key transcriptional factors (CRX, RAX, NEURO-D, OTX2) allows the direct differentiation of fibroblasts to retinal photoreceptor cells. This reprogramming was achieved with a combination of mono-cistronic viruses. Although the combination of mono-cistronic viruses was useful, a relatively high titer of recombinant viruses was necessary because co-infections are required. To overcome this issue, we established a poly-cistronic expression system for direct reprogramming and analyzed the biological characteristics of introduced cells after the exogenous introduction. The coding region of four reprogramming factors and EGFP (CRX, RAX, NEURO-D, OTX2, and EGFP; CNROE) was inserted into multiple sites of the pMYs-IP retrovirus or CSII-CMV lentivirus vector. The recombinant viruses were exposed to HE16 human embryonic fibroblasts. The expression levels of cone related genes were detected with real-time PCR. We detected the activation of two of the photoreceptor-related genes after the poly-cistronic expression of CRX, RAX, NEURO-D, and OTX2, but the rest of the genes did not exhibit transcriptional elevation. We concluded that the poly-cistronic expression of CNROE induced partial reprogramming into photoreceptor cells. We hypothesize that the direct reprogramming into photoreceptor cells might require relatively high protein expression levels of transcriptional factors.


A safe, simple and cost-effective protocol for blood transfusion in primary total knee replacement.
Patients undergoing total knee replacement (TKR) in the UK usually have either blood cross-matched or have an auto-transfusion of drained blood postoperatively. A previous retrospective audit of blood requirements in patients who had undergone primary TKR showed that a large amount of cross-matched blood was wasted as the CT ratio (ratio of number of units of blood cross-matched to number of units transfused) of 4.9:1 was obtained. The range recommended by the Blood Transfusion Society is 2:1 to 3:1. A protocol was introduced to group and save plus antibody screen for all patients and to cross-match 2 units of blood pre-operatively in patients with either a haemoglobin of less than 12.5 g/dl or with multiple red cell antibodies in their blood. The trigger point for blood transfusion postoperatively was also reduced from 9.0 g/dl to 8.0 g/dl, unless the patient was clinically symptomatic. A further prospective study involving 50 patients was carried out using the new protocol. Five patients required cross-matching pre-operatively, three with haemoglobin less than 12.5 g/dl and two with multiple red cell antibodies. Postoperatively, the patients with haemoglobin of less than 12.5 g/dl required blood transfusion of 2 units each, reducing the CT ratio to 1.7:1. The patients with red cell antibodies did not require a blood transfusion. The benefits from above protocol are 2-fold: patient safety, as risks of transfusion are avoided; and cost saving, in regards to haematology technician time and auto-transfusion sets which cost around pound 70 each.