T cell clone producing an IL 1-like activity after stimulation by antigen-presenting B cells.
We have analyzed the involvement of IL 1 in a cellular interaction during which murine B cells serve as antigen-presenting cells for a T cell clone (D10.G4.1). This T cell clone was chosen for study because it is highly dependent on exogenous IL 1 for concanavalin A (or anti-receptor antibody)-induced proliferation. We observed that B cells presented either self I-Ak plus nominal antigen or allogeneic I-Ab, and consequently induced proliferation of this T cell clone without any requirement for exogenous IL 1. After an overnight co-culture with allogenic B cells, we detected an IL 1-like activity in lysates of the cell mixture, but were unable to detect a similar activity in the culture medium. This IL 1-like activity was induced in a MHC-restricted manner. We determined that this intracellular IL 1-like activity was derived from the D10.G4.1-responding T cells, rather than from the antigen-presenting B cells. Moreover, we were unable to detect an IL 1-like activity in murine B cells after a variety of stimuli including LPS, anti-Ig, activated T cell supernatants, or combinations of these stimuli. Hybridization of cytoplasmic RNA with an oligonucleotide probe for human IL 1-alpha confirmed the T cell source of the biological activity. The IL 1-like factor derived from the cell culture mixture lysate showed two broad peaks of activity after gel filtration, one in the 10,000 to 20,000 dalton range and the second in the 35,000 to 45,000 dalton range. This first description of IL 1 activity produced by a T cell may introduce a new element to the early events leading to T cell activation and proliferation.


Morbidity and mortality of orthostatic hypotension: implications for management of cardiovascular disease.
Orthostatic hypotension (OH) is the failure of cardiovascular reflexes to maintain blood pressure on standing from a supine or sitting position. Although OH may cause symptoms of dizziness or syncope, asymptomatic OH (AOH) is far more common and is an independent risk factor for mortality and cardiovascular disease (CVD). The prevalence of AOH increases with age, the presence of hypertension or diabetes and the use of antihypertensive or other medications. The implications of AOH for the treatment of CVD and hypertension are not well defined. This review provides an overview of the current information on this topic and recommends the more frequent assessment of OH in clinical practice and in future clinical trials.


Microbial metabolomics: toward a platform with full metabolome coverage.
Achieving metabolome data with satisfactory coverage is a formidable challenge in metabolomics because metabolites are a chemically highly diverse group of compounds. Here we present a strategy for the development of an advanced analytical platform that allows the comprehensive analysis of microbial metabolomes. Our approach started with in silico metabolome information from three microorganisms-Escherichia coli, Bacillus subtilis, and Saccharomyces cerevisiae-and resulted in a list of 905 different metabolites. Subsequently, these metabolites were classified based on their physicochemical properties, followed by the development of complementary gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry methods, each of which analyzes different metabolite classes. This metabolomics platform, consisting of six different analytical methods, was applied for the analysis of the metabolites for which commercial standards could be purchased (399 compounds). Of these 399 metabolites, 380 could be analyzed with the platform. To demonstrate the potential of this metabolomics platform, we report on its application to the analysis of the metabolome composition of mid-logarithmic E. coli cells grown on a mineral salts medium using glucose as the carbon source. Of the 431 peaks detected, 235 (=176 unique metabolites) could be identified. These include 61 metabolites that were not previously identified or annotated in existing E. coli databases.


IL-7 receptor expression provides the potential for long-term survival of both CD62Lhigh central memory T cells and Th1 effector cells during Leishmania major infection.
Infection with the intracellular protozoan parasite Leishmania major induces a state of concomitant immunity wherein secondary immunity is dependent upon the persistence of the original pathogen. Our laboratory has described two populations of Leishmania-induced CD4(+) T cells that contribute to immunity: CD62L(high) central memory T (T(CM)) cells and CD62L(low) effector T cells. To determine whether the prosurvival cytokine IL-7 contributes to maintaining these T cells, we examined expression of the IL7R on CD4(+) T cells activated during L. major infection. We found that T(CM) cells present in chronically infected mice expressed high levels of the IL7R. However, in addition to the expression of the IL7R by T(CM) cells, CD62L(low) cells responding to L. major infection expressed the IL7R. Additional experiments revealed that a large percentage of the IL7R(high)CD62L(low) cells were Th1 cells, based on transcription at the IFN-gamma locus and up-regulation of the Th1-promoting transcription factor T-bet. The up-regulation of T-bet did not prevent IL7R expression by L. major-responding CD4(+) T cells, nor did the absence of T-bet result in increased IL7R expression. Finally, blockade of IL7R signaling decreased the number of T-bet(+)CD4(+) T cells, reduced IFN-gamma production, and inhibited delayed-type hypersensitivity responses in immune mice challenged with L. major, indicating that IL7R signaling contributes to the maintenance of Th1 effector cells. Thus, both T(CM) and Th1 effector cells can express the IL7R during chronic L. major infection, which provides a potential means for their long-term survival in addition to the presence of persisting parasites.


Novel gene therapeutic strategies for neurodegenerative diseases.
The convergent pathobiologic model of Parkinson's disease stipulates that disparate insults initiate a disease process that obligately share a common pathway leading to cell death. A combinatorial treatment which targets various steps in this pathway is likely to be the most successful therapeutic strategy. As advances are made in the field of neuroimaging and pharmacogenomics, early detection of sporadic PD will become a reality. Early intervention will likely spare more dopaminergic neurons and extend the quality of life for the patient. Continued advancements in the fields of pharmacology, neurosurgery, and gene therapy will strengthen the armamentarium available for the treatment of PD patients.


The population biology of transposable elements.
A transposable element can be defined as a DNA sequence capable of moving to new sites in the genome. Such DNA sequences have been described in a wide range of organisms. The evolutionary processes affecting transposable elements can thus be divided into two categories: changes in sequence and changes in genomic location. As with other types of evolutionary change, the nature of the evolutionary process will be reflected in the extent and type of genetic variation existing in wild populations. Quantitative models of the evolution of transposable element sequences and positions will be outlined, and related to relevant data. The extent to which models designed to describe obvious transposable elements such as the mobile sequences of Drosophila are also applicable to interspersed repetitive DNAs from other species will be discussed.


Maintenance of a somatotopic cortical map in the face of diminishing thalamocortical inputs.
This study addresses the extent of divergence in the ascending somatosensory pathways of primates. Divergence of inputs from a particular body part at each successive synaptic step in these pathways results in a potential magnification of the representation of that body part in the somatosensory cortex, so that the representation can be expanded when peripheral input from other parts is lost, as in nerve lesions or amputations. Lesions of increasing size were placed in the representation of a finger in the ventral posterior thalamic nucleus (VPL) of macaque monkeys. After a survival period of 1-5 weeks, area 3b of the somatosensory cortex ipsilateral to the lesion was mapped physiologically, and the extent of the representation of the affected and adjacent fingers was determined. Lesions affecting less than 30% of the thalamic VPL nucleus were without effect upon the cortical representation of the finger whose thalamic representation was at the center of the lesion. Lesions affecting about 35% of the VPL nucleus resulted in a shrinkage of the cortical representation of the finger whose thalamic representation was lesioned, with concomitant expansion of the representations of adjacent fingers. Beyond 35-40%, the whole cortical representation of the hand became silent. These results suggest that divergence of brainstem and thalamocortical projections, although normally not expressed, are sufficiently great to maintain a representation after a major loss of inputs from the periphery. This is likely to be one mechanism of representational plasticity in the cerebral cortex.


Local wavelet-vaguelette-based functional classification of gene expression data.
This paper focuses on the problem of functional statistical classification of gene expression curves. A local-wavelet-vaguelette-based functional logistic regression approach is presented. This approach is specially suitable for the classification of non-stationary singular (non-differentiable) curves. The performance of the methodology proposed is illustrated by implementing it for the classification of yeast cell-cycle temporal gene expression profiles. A simulation study is also carried out for comparison with other functional classification methodologies.


High-pressure on-chip mechanical valves for thermoplastic microfluidic devices.
A facile method enabling the integration of elastomeric valves into rigid thermoplastic microfluidic chips is described. The valves employ discrete plugs of elastomeric polydimethylsiloxane (PDMS) integrated into the thermoplastic substrate and actuated using a threaded stainless steel needle. The fabrication process takes advantage of poly(ethylene glycol) (PEG) as a sacrificial molding material to isolate the PDMS regions from the thermoplastic flow channels, while yielding smooth contact surfaces with the PDMS valve seats. The valves introduce minimal dead volumes, and provide a simple mechanical means to achieve reproducible proportional valving within thermoplastic microfluidic systems. Burst pressure tests reveal that the valves can withstand pressures above 12 MPa over repeated open/close cycles without leakage, and above 24 MPa during a single use, making the technology well suited for applications such as high performance liquid chromatography. Proportional valve operation is demonstrated using a multi-valve chemical gradient generator fabricated in cyclic olefin polymer.


Correlation of Serum Uric Acid Levels with Nonculprit Plaque Instability in Patients with Acute Coronary Syndromes: A 3-Vessel Optical Coherence Tomography Study.
Elevated serum uric acid (SUA) level is known to be a prognostic factor in patients with acute coronary syndrome (ACS). However, the correlation between SUA level and coronary plaque instability has not been fully evaluated. The aim of this study was to investigate the association between SUA level and plaque instability of nonculprit lesions in patients with ACS using optical coherence tomography. A total of 150 patients with ACS who underwent 3-vessel optical coherence tomography were selected. Patients were classified into 3 groups according to tertiles of SUA level. There was a trend towards a thinner fibrous cap (0.15 ± 0.06 versus 0.07 ± 0.01 versus 0.04 ± 0.01 mm2, p < 0.001) and a wider mean lipid arc (169.41 ± 33.16 versus 177.22 ± 37.76 versus 222.43 ± 47.65°, p < 0.001) with increasing SUA tertile. The plaques of the high and intermediate tertile groups had a smaller minimum lumen area than the low tertile group (6.02 ± 1.11 versus 5.38 ± 1.28 mm2, p < 0.001). In addition, thin-cap fibroatheromas, microvessels, macrophages, and cholesterol crystals were more frequent in the high tertile group than the low and intermediate groups. Multivariate analysis showed SUA level to be a predictor of plaque instability.


[Ventricular assist device].
There are 2 kinds of artificial heart used for clinical patients with end-stage heart failure. One is a ventricular assist device (VAD) and the other is a total artificial heart (TAH). Only paracorporeal VADs are commercially available currently in Japan, such as Toyobo VAD and BVS 5000. Paracorporeal Zeon VAD and implantable Novacor LVAD were approved by the MHLW in the past but already disappeared from the Japanese market in 2005 and 2006. A little more than 1,000 LVADs had been implanted since 1980 in Japan until last autumn, while no TAH has been implanted in patients. In the world 1st generation pulsatile implantable LVADs have been taken over by the 2nd and 3rd generation non-pulsatile implantable LVADs in the past 10 years. In Japan 4 kinds of implantable LVADs are under clinical evaluation, HeartMate XVE (1st generation, HeartMate VE was used in the Japanese clinical trial), Jarvik 2000 (2nd generation), Evaheart (2nd generation), and DuraHeart (3rd generation). All 5 patients with HeartMate VE survived more than 1 year during the clinical trial, 13 patients with Evaheart were long survivors or successfully heart-transplanted. Although follow-up interval is relatively short, all 6 patients with DuraHeart and 5 of the 6 patients with Jarvik 2000 recovered from end-stage heart failure and are keeping very good condition. Most of them returned to their home and are waiting for heart transplantation. Especially the 3rd generation of non-pulsatile LVADs is expected to have long-term durability and less frequency of thromboembolic episode.


Sustained-release metoclopramide plus methylprednisolone versus placebo plus methylprednisolone as antiemetic prophylaxis during non-cisplatin chemotherapy. A randomized double-blind cross-over trial.
In a randomized double-blind cross-over trial, sustained-release metoclopramide (S) plus methylprednisolone (M) was compared with placebo (P) plus methylprednisolone as antiemetic prophylaxis during two cycles of non-cisplatin chemotherapy. S was administered as 60 mg every 12 h commencing on the evening before chemotherapy up to total of 300 mg metoclopramide in 2.5 days. Evaluation of nausea and vomiting was done by self-assessment schemes and visual analog scales. Fifty patients were included and 36 fulfilled both cycles. Mild nausea and vomiting were experienced by 81% and 83% in the S + M and P + M groups, respectively, while 42% and 39% showed complete control of nausea and vomiting during the first day of treatment. Moderate-dose S did not add to the antiemetic efficacy of M in non-cisplatin chemotherapeutic regimens.


Relaxometric and solution NMR structural studies on ditopic lanthanide(III) complexes of a phosphinate analogue of DOTA with a fast rate of water exchange.
A novel "ditopic" ligand containing two monophosphinate triacetate DOTA-like units linked by a thiourea bridge has been synthesized and its complexes with Ln3+ ions (Ln = Y, Eu, Gd, Dy) investigated by NMR spectroscopy and relaxometry. The presence of one water molecule in the first coordination sphere has been determined by the measurement of the dysprosium(III)-induced 17O NMR shifts. The 1H and 31P NMR spectra of the Eu(III) derivative indicate a higher abundance of the fast-exchanging twisted square antiprismatic (m) isomer than the isomeric square antiprismatic (M; m/M = 3:2) complex. The analysis of the 89Y and 13C T1 NMR relaxation times in the Gd(III)/Y(III) mixed complex have provided useful structural information. Values of ca. 6.3 and 8.2 A for the Gd...Y and Gd...C distances, respectively, have been estimated which indicate a rather compact solution structure. This result finds support in the value of the relaxivity whose increase (at 20 MHz and 298 K) on passing from the monomeric (5.7 s(-1) mM(-1)) to the ditopic complex (8.2 s(-1) mM(-1)) can be attributed to the doubling of the inner-sphere term following the doubling of the molecular size. The structural and dynamic relaxivity-controlling parameters were assessed by a simultaneous fitting of the variable temperature 17O NMR and 1H NMRD relaxometric data. The mean water residence lifetime (298tauM) has been found to be 53 ns, one of the shortest values reported for ditopic complexes. The reorientational correlation time is two times longer (298tauR = 183 ps) than the corresponding value of the parent monomeric Gd(III) complex, thus supporting the view of a limited degree of internal rotation. The possible influence of magnetic Gd-Gd coupling has been excluded by a comparison of the 1H NMRD profiles of the homodinuclear Gd(III)/Gd(III) and the heterodinuclear Gd(III)/Y(III) complexes.


An electrophysiological study of peripheral polyneuropathy in brucellosis.
Brucellosis is an anthropozoonotic disease with heterogeneous clinical presentations. This study aims to investigate the peripheral nervous system (PNS) involvement in brucellosis. A total of 57 patients with brucellosis, and 42 age- and gender-matched healthy subjects were enrolled into the study. We performed motor conduction studies that included bilateral median, ulnar, tibial and peroneal nerves, and sensory nerve investigations from bilateral median, ulnar, radial, sural and medial plantar nerves. Among patients with brucellosis, 21 had neuropathic symptoms. Of these, 9 had abnormalities in nerve conduction studies. The electromyographic testing revealed abnormalities in 2 patients without neuropathic symptoms. Overall, 11 patients (4 males, 7 females) with brucellosis (19.3%) had polyneuropathy (PNP). The mean age of patients with PNP was 52.63 ± 19.06 years, being significantly higher than those without PNP (P = 0.006). The mean duration of brucellosis was also longer in patients with PNP, but not significant. The mean distal latency (DL) and nerve conduction velocity (NCV) values were almost always longer in patients with brucellosis than controls, though not statistically significant. Our results showed that brucellosis causes clinical or subclinical peripheral PNP, and should therefore be considered as a cause of PNP, especially in endemic regions.


Design, methodology and baseline data of a school-based cohort study in Central China: the Anyang Childhood Eye Study.
To report the design, methods and baseline data of the Anyang Childhood Eye Study (ACES), aiming to determine the prevalence, incidence and risk factors of myopia and other ocular diseases in children in central China. The ACES was a school-based cohort study conducted in Anyang city. Students have been examined and will be followed-up annually for 3-5 years. Ocular examinations included visual acuity, identification of amblyopia and strabismus, ocular biometry, optical coherence tomography, retinal photography, cycloplegic autorefraction, accommodative response, peripheral refraction, visual perception and so on. A questionnaire survey was performed to collect risk factors for myopia including near work, outdoor activity, parental myopia, birth history, habits of wearing spectacles and reading, living habits, food habits and so on. Of 3112 grade 1 and 2363 grade 7 students eligible for the ACES, 2893 (93.0%) and 2267 (95.9%) participated in the study, with a mean age of 7.1 years (range 5.7-9.3 years) and 12.7 years (range 10.0-15.9 years), and proportions of male to be 57.8% and 50.0%, respectively. The prevalence rates of myopia, high myopia, emmetropia and hyperopia were 3.9%, 0.1%, 72.9% and 23.3% in grade 1, and 67.3%, 2.7%, 31.4% and 1.2% in grade 7, respectively. Similar data were found in the children confined in 6-year-old and 12-year-old ages. The ACES is the first large-scale cohort study in China with baseline response rates over 90%. Continuous documentation of changes and risk factors of refractive errors in these cohorts would provide new insights into myopia control in school-aged children.


The computer simulation of RNA folding involving pseudoknot formation.
The algorithm and the program for the prediction of RNA secondary structure with pseudoknot formation have been proposed. The algorithm simulates stepwise folding by generating random structures using Monte Carlo method, followed by the selection of helices to final structure on the basis of both their probabilities of occurrence in a random structure and free energy parameters. The program versions have been tested on ribosomal RNA structures and on RNAs with pseudoknots evidenced by experimental data. It is shown that the simulation of folding during RNA synthesis improves the results. The introduction of pseudoknot formation permits to predict the pseudoknotted structures and to improve the prediction of long-range interactions. The computer program is rather fast and allows to predict the structures for long RNAs without using large memory volumes in usual personal computer.


The Defense of My Aging Self: A Report From the Field.
This article offers a field report-experiences, thoughts, and anecdotes-of my personal passage into older adulthood and my attempts to maintain a healthy sense of personhood in the context of the normative ageism in my world. Specifically, I illustrate how my career-long engagement with the scientific base of gerontology-particularly life-span developmental and social psychology-has been a key ally in informing my aging and in assisting my defense against internalization of negative stereotypes. Also, I offer some thoughts drawn from personal life experience that may inform research and application efforts designed to modify some of the psychological motives that sustain ageism in our shared environment.


[Influence of imaging parameters on the measurement of apparent diffusion coefficient].
The apparent diffusion coefficient (ADC) values are calculated by using signal intensity in diffusion-weighted images (DWIs) with two or more different b-value. Therefore, the signal to noise ratio (SNR) of DWI with higher b-value may have a big influence on the measured ADC value. We examined the influence of the imaging parameters on the calculated ADC values. The SNR of DWI increased by using a larger voxel size, by means of a decreased number of matrix, an increased slice thickness, and an increased field of view (FOV). However, when the number of excitations was increased to improve the SNR of DWI, the signal intensity of background noise was observed to be slightly increased. It was suggested that the consistency of measured ADC was not preserved when the signal of the DWIs with higher b-value dropped close to the noise level.


[Thymic carcinoma; analysis of nine clinical studies].
Nine cases of thymic carcinoma (mean age 53.9 years old, 8 males and 1 female) were treated in our hospital from 1973 to 1998. The histological subtypes of thymic carcinoma were squamous cell in six, adenosquamous cell in one, Lymphoepithelioma-like carcinoma in one, mixed type of small cell and squamous cell in one. We applied Yamakawa clinical staging, 9 cases consisted of 2 stage II cases, 2 stage III cases, and 5 stage IV b cases. Adjuvant radiotheraphy was administered in one case, registering remarkable reduction of tumor size. All of the four cases which had been performed complete resection are alive without recurrence for 17 to 136 months after operation. The mean survival time was 47 months in stage III, and 18 months in stage IV b. By the review of the Japanese literature, most of the patient who revealed long-term survival fulfill the conditions as follow, squamous cell carcinoma in histologically, earlier stage than III at his operation, performed complete resection, performed postoperative radiotherapy. Our cases covering all these condition are alive without carcinoma for 136 months after operation.


Mortality after discharge from acute care hospitalization with traumatic brain injury: a population-based study.
Ventura T, Harrison-Felix C, Carlson N, DiGuiseppi C, Gabella B, Brown A, DeVivo M, Whiteneck G. Mortality after discharge from acute care hospitalization with traumatic brain injury: a population-based study. To characterize mortality after acute hospitalization with traumatic brain injury (TBI) in a socioeconomically diverse population. Population-based retrospective cohort study. Statewide TBI surveillance program. Colorado residents with TBI discharged alive from acute hospitalization between 1998 and 2003 (N=18,998). Not applicable. Vital status at the end of the study period (December 31, 2005) and statewide population mortality rates were used to calculate all-cause and cause-specific standardized mortality ratios (SMRs) and life expectancy compared with population mortality rates. The influence of demographics, injury severity, and comorbid conditions on time until death was investigated using age-stratified Cox proportional hazards modeling. Patients with TBI carried about 2.5 times the risk of death compared with the general population (SMR=2.47; 95% confidence interval [CI], 2.31-2.65). Life expectancy reduction averaged 6 years. SMRs were largest for deaths caused by mental/behavioral (SMR=3.84; 95% CI, 2.67-5.51) and neurologic conditions (SMR=2.79; 95% CI, 2.07-3.77) and were smaller but significantly higher than 1.0 for an array of other causes. Injury severity and older age increased mortality among young people (age <20y). However, risk factors for mortality among adults age 20 and older involved multiple domains of demographics (eg, metropolitan residence), injury-related measures (eg, falls versus vehicular incidents), and comorbidity (eg, > or =3 comorbid health conditions versus none). TBI confers an increased risk of mortality in the months and years after hospital discharge. Although life expectancy is reduced across the population, the excess in mortality lessens as time since injury increases. Specific risk factors (eg, high injury severity, poor general health) pose an especially high threat to survival and should prompt an increased vigilance of health status, especially among younger patients.


Arf1-GTP-induced tubule formation suggests a function of Arf family proteins in curvature acquisition at sites of vesicle budding.
ADP-ribosylation factor (Arf) and related small GTPases play crucial roles in membrane traffic within the exo- and endocytic pathways. Arf proteins in their GTP-bound state are associated with curved membrane buds and tubules, frequently together with effector coat proteins to which they bind. Here we report that Arf1 is found on membrane tubules originating from the Golgi complex where it colocalizes with COPI and GGA1 vesicle coat proteins. Arf1 also induces tubulation of liposomes in vitro. Mutations within the amino-terminal amphipathic helix (NTH) of Arf1 affect the number of Arf1-positive tubules in vivo and its property to tubulate liposomes. Moreover, hydrophilic substitutions within the hydrophobic part of its NTH impair Arf1-catalyzed budding of COPI vesicles in vitro. Our data indicate that GTP-controlled local induction of high curvature membranes is an important property of Arf1 that might be shared by a subgroup of Arf/Arl family GTPases.


Transmesenteric hernia with two mesenteric defects in an adult.
Transmesenteric hernia is a rare cause of small bowel strangulation in adults and, to our knowledge, no one has reported the existence of two mesenteric defects in an adult. Our patient was a 73-year-old Japanese woman who presented to our emergency department complaining of abdominal pain and nausea. Computed tomography with contrast enhancement revealed a closed loop obstruction in the pelvis, suggesting small bowel strangulation due to an internal hernia. The emergency exploratory laparotomy indicated a small bowel strangulation caused by a transmesenteric hernia. With the examination across whole parts of the mesentery, we identified another small defect. Both defects were closed by suture intraoperatively, and the patient's postoperative course was satisfactory. Searching for whole parts of the mesentery after the reduction of a hernia can help prevent the recurrence of internal hernias.


Fruit volatilome profiling through GC × GC-ToF-MS and gene expression analyses reveal differences amongst peach cultivars in their response to cold storage.
Peaches have a short shelf life and require chilling during storage and transport. Peach aroma is important for consumer preference and determined by underlying metabolic pathways and gene expression. Differences in aroma (profiles of volatile organic compounds, VOCs) have been widely reported across cultivars and in response to cold storage. However, few studies used intact peaches, or used equilibrium sampling methods subject to saturation. We analysed VOC profiles using TD-GC × GC-ToF-MS and expression of 12 key VOC pathway genes of intact fruit from six cultivars (three peaches, three nectarines) before and after storage at 1 °C for 7 days including 36 h shelf life storage at 20 °C. Two dimensional GC (GC × GC) significantly enhances discrimination of thermal desorption gas chromatography time-of-flight mass spectrometry (TD-GC-ToF-MS) and detected a total of 115 VOCs. A subset of 15 VOCs from analysis with Random Forest discriminated between cultivars. Another 16 VOCs correlated strongly with expression profiles of eleven key genes in the lipoxygenase pathway, and both expression profiles and VOCs discriminated amongst cultivars, peach versus nectarines and between treatments. The cultivar-specific response to cold storage underlines the need to understand more fully the genetic basis for VOC changes across cultivars.


Elevated plasma ferritin is associated with increased incidence of type 2 diabetes in middle-aged and elderly Chinese adults.
Epidemiological studies suggest that elevated circulating ferritin is associated with heightened incident diabetes in mainly Western populations, although the results were not entirely consistent. We aimed to prospectively investigate the ferritin-diabetes association in an Asian population for the first time, to our knowledge, and also to examine this association with an updated meta-analysis. Our prospective study included 2198 community-living Chinese between 50 and 70 y of age in 2005. All individuals participated in a 6-y follow-up survey in 2011. Fasting plasma ferritin, high-sensitivity C-reactive protein (hsCRP), adiponectin, and γ-glutamyltransferase (GGT) were measured at baseline. A total of 538 incident diabetes cases were documented by self-reports and/or fasting glucose ≥7.0 mmol/L at the follow-up survey. After multiple adjustments, the RR of type 2 diabetes was 1.90 (95% CI: 1.37, 2.65) when comparing the highest with the lowest sex-specific ferritin quintile. The association remained significant after further controlling for BMI, hsCRP, adiponectin, and GGT. To update the evidence reported in previous meta-analyses, we searched all prospective studies evaluating the association between blood ferritin and incident diabetes on PubMed prior to October 24, 2012. Besides our prospective study, 9 additional studies were also included. The pooled RR was 1.60 (95% CI: 1.25, 2.04) when comparing the highest with the lowest category of ferritin with a moderate heterogeneity (I(2) = 49.0%; P = 0.03). A significant linear dose-response relationship was detected in this meta-analysis. Overall, our results indicate an independent and significant positive association between higher plasma ferritin, a marker of elevated body iron stores, and increased risk of developing type 2 diabetes in middle-aged and elderly Chinese adults, which is similar to Western populations.


Human X-box-binding protein 1 is required for the transcription of a subset of human class II major histocompatibility genes and forms a heterodimer with c-fos.
A complementary DNA encoding a member of the leucine-zipper class of proteins (human X-box-binding protein, hXBP-1) that binds to the 3' end of the conserved X box (X2) of the HLA-DRA major histocompatibility complex gene was recently described. Further gel-retardation analysis has demonstrated that hXBP-1 also binds to HLA-DPB X2 but not to other X2 sequences. Transient transfection of a mammalian expression vector with the hXBP-1 cDNA inserted in the antisense orientation represses the surface expression of HLA-DR and HLA-DP in Raji cells. Cotransfection of the antisense hXBP-1 vector with a HLA-DRA/chloramphenicol acetyltransferase (but not a HLA-DQB/chloramphenicol acetyltransferase) reporter plasmid decreases chloramphenicol acetyltransferase activity in Raji cells and in gamma-interferon-treated HeLa cells relative to cells cotransfected with a control antisense vector. Moreover, hXBP-1 is shown to form a stable heterodimer with the product of the c-fos protooncogene. These data suggest that the hXBP-1 c-fos heterodimer is critical for the transcription of a subset of the human class II major histocompatibility complex genes and that the regulatory mechanisms for the different class II genes are distinct.


Experimental Evaluation of a Game-Theoretic Human Driver Steering Control Model.
Automated vehicle steering control systems have great potential to improve road safety. The development of such systems calls for mathematical driver models able to represent human drivers' steering behavior in response to automated steering intervention. This article concerns the experimental evaluation of a game-theoretic driver steering control model. The driver model centers on a steering control strategy developed based on the Nash equilibrium of a theoretic noncooperative game between the driver and automated steering controller. The key parameters of the game-theoretic driver model are identified by fitting the model to real driver steering behavior measured from six driver subjects in an experiment using a driving simulator. The game-theoretic driver model is evaluated by compared to a "conventional" optimal-control-theoretic driver model, and analyzing their model fitting errors. Results from the analysis demonstrate that the game-theoretic driver model is statistically significantly better than the conventional driver model for representing three out of the six subjects' steering behavior. For the other three subjects, both the two models perform statistically equivalently well.


Brain and spinal manifestations of Miller-Dieker syndrome.
A 6-month-old infant with LIS1 17p13.3 deletion-positive Miller-Dieker syndrome (MDS) presented with increased seizures in the setting of a Pseudomonal and Enterococcal urinary tract infection and a buttock abscess associated with a lumbosacral dermal sinus tract. MRI of the neuraxis revealed lissencephaly (figure 1), a tethered cord without lipoma or other mass (figure 2A), and an infected lumbosacral dermal sinus tract. Communication with the spinal canal could not be appreciated (figure 2B). The dermal sinus was explored and found not to extend into the spinal canal. This tract was excised and the lateral abscess drained. Tethered cord release is planned upon resolution of infection.


Subclinical rhythmic electrographic discharge in adults: an atypical evolution.
The subclinical rhythmic electrographic discharge in adults (SREDA) has been rarely reported. We describe the case of a 71 year-old man with transient ischemic attack, whose EEG showed an atypical evolution of this uncommon pattern. The computed tomography scan and cerebrospinal fluid were normal. We are not aware of any other report in the literature with this atypical evolution of SREDA.


Differentiated-Rate Clean Heating Strategy with Superior Environmental and Health Benefits in Northern China.
Residential heating using solid fuels contributes significantly to air pollution and has subsequent health impacts in China. To mitigate emissions, a clean heating campaign (CHC-1) covering 28 municipalities has been implemented. Although only a single penetration rate was initially planned by CHC-1 for all municipalities, outcomes in the different municipalities varied considerably. Recently, a second phase (CHC-2) has been launched for the remaining 128 municipalities in northern China with once again a fixed penetration rate set. Here, we quantified factors that affected the penetration rates of CHC-1, developed an intervention scheme with differentiated targets for CHC-2, and compared the environmental and health benefits of the fixed- and differentiated-rate strategies. We found that the penetration rates of CHC-1 depended on per capita income, terrain slope, and population density and that such relationships could be quantified using a piecewise regression model. This model was applied to develop a differentiated-rate strategy for CHC-2. It clearly evidenced that a differentiated scheme would be more environmentally beneficial. Although the same number of rural households can achieve clean heating under both intervention scenarios, the proposed differentiated strategy can prevent 30 000 (23 000-34 000) premature deaths associated with residential heating annually compared to the 26 000 (21 000-31 000) premature deaths prevented under the fixed-rate scheme. Differences among gender and age groups and the effects of urbanization and aging are also discussed.


Single effects of apolipoprotein B, (a), and E polymorphisms and interaction between plasminogen activator inhibitor-1 and apolipoprotein(a) genotypes and the risk of coronary artery disease in Czech male caucasians.
To evaluate whether polymorphisms in genes whose products are involved in lipid metabolism and fibrinolysis alter the risk of coronary artery disease (CAD), allele frequencies of four genetic polymorphisms were ascertained by PCR-based methods in 175 Czech male patients with coronary artery disease and in 222 Czech men with no symptoms of CAD. The following polymorphisms were studied: apolipoprotein B (apo B) signal peptide insertion/deletion polymorphism, 5' apolipoprotein(a) [apo(a)] TTTTA repeat polymorphism, apolipoprotein E (apo E) varepsilon2, varepsilon3, varepsilon4 polymorphism, and plasminogen activator inhibitor-1 (PAI-1) 4G/5G promoter polymorphism. Apo B and apo(a) allele frequencies differed significantly between the CAD and the control groups (P<0.01 each), with higher frequencies of apo B deletion and apo(a) shorter repeat alleles in the CAD group. We did not observe any differences in allele frequencies of either PAI-1 or apo E polymorphisms but the genotype frequencies of apo E were slightly different between the two groups (P<0.05). In addition, we observed a gene-gene interaction between the PAI-1 and apo(a) polymorphisms with respect to the risk of CAD. None of the polymorphisms studied were associated with the severity of CAD or a history of myocardial infarction. Our findings support the idea that several polymorphisms in apolipoprotein genes may by themselves and/or in interaction with other polymorphisms contribute to risk factors for CAD in men.


The Attitudes to Disability Scale (ADS): development and psychometric properties.
This paper describes the development of an Attitudes to Disability Scale for use with adults with physical or intellectual disabilities (ID). The aim of the research was to design a scale that could be used to assess the personal attitudes of individuals with either physical or ID. The measure was derived following standard WHOQOL methodology as part of an international trial. In the pilot phase of the study, 12 centres from around the world carried out focus groups with people with physical disabilities, people with ID, with their carers, and with relevant professionals in order to identify themes relevant for attitudes to disability. Items generated from the focus groups were then tested in a pilot study with 1400 respondents from 15 different centres worldwide, with items being tested and reduced using both classical and modern psychometric methods. A field trial study was then carried out with 3772 respondents, again with the use of both classical and modern psychometric methods. The outcome of the second round of data collection and analysis is a 16-item scale that can be used for assessment of attitudes to disability in physically or intellectually disabled people and in healthy respondents. The Attitudes to Disability Scale is a new psychometrically sound scale that can be used to assess attitudes in both physically and intellectually disabled groups. The scale is also available in both personal and general forms and in a number of different language versions.


Consequences of age-related splanchnic sequestration of leucine on interorgan glutamine metabolism in old rats.
Dietary leucine (Leu) serves as a nitrogen donor for de novo glutamine (Gln) synthesis in muscle. However, aging is characterized by an increase in the splanchnic extraction of Leu (SPELeu), i.e., splanchnic sequestration (SSLeu), which may affect muscle Gln metabolism and its subsequent homeostasis at the whole-body level. The aim of the work was to assess the effect of age-related SSLeu on Gln metabolism in the muscle, gut, liver, kidney, and Gln exchanges among these organs during fed conditions. Young-adult (3-mo-old) or aged (24-mo-old), male Sprague-Dawley rats were studied during fed condition [infusion of amino acids (AA) into the duodenum from time 0 min (T0) to T60] under anesthesia. L-[5-(15)N]Gln and L[1-(13)C]Leu were infused into the jugular vein and L-[5,5,5-(2)H3]Leu into the duodenum. At T60, blood samples were taken from carotid artery, portal vein, hepatic vein, renal vein, and inferior vena cava for tracer-tracee ratio and AA level measurements. SSLeu was observed in old rats and was negatively correlated with muscle Gln production (r = -0.501, P < 0.01). In addition, reduced Gln muscle release in old rats was accompanied by reduced Gln uptake by the gut and kidney. However, net Gln balance across organs was not different between young adult and old rats. During fed conditions in old rats, muscle Gln production and release are reduced in relation to the observed, increased SPELeu and reduced renal and intestinal Gln uptake to maintain whole-body Gln homeostasis. Our results demonstrate the existence of an age-related change of interorgan Gln metabolism, which may be, in part, driven by SSLeu.


The role of interstitial native defects in the topological insulator Bi2Se3.
Bi2Se3 is a prominent narrow gap semiconductor with a rhombohedral crystal structure and potential applications in thermoelectric and spintronic technologies. Its electrical conduction is ruled by native point defects inducing an n-type degenerate behavior. Here, we present a first principles study of the point defects in Bi2Se3, focusing on the relevance of the interstitial sites. A density functional methodology was employed with van der Waals correction and spin-orbit coupling in order to achieve a better description of the defects. The results indicate that interstitial Bi atoms in octahedral sites between two consecutive quintuple layers have a lower formation energy than selenium vacancies and that these interstitials could act as a possible source of free electron carriers. In addition, we show that the utilization of an experimental or strained lattice constant in the calculations may lead to an under- or overestimation of the defect formation energies.


Meeting the special medicinal needs of patients.
The clinical needs of patients can often be met by prescribing and administering a licensed medicine. District nurses will have some cases where a medicine cannot be used because the patient is unable to take it in its licensed form. In such situations, district nurses may be able to arrange for the patient to be supplied with an unlicensed medicine specifically made to meet that patient's special need. This article discusses the options available to district nurses to lawfully supply patients who have a special medication need due to, for example, an allergy to a particular ingredient or an inability to swallow solid oral medicines, such as tablets or capsules.


[Invasion of fibrosarcoma cells transfected with PAI-2 gene].
The effect of PAI-2 on the invasion of fibrosarcoma cells in vitro and in vivo was investigated. The control cells (C+, C+ pem) and PAI-2 transfectants (C+ exp) were used in the assay of the degradation of isotopically labelld 3H-ECM and were injected sc into athymic/nude mice. Recombinant PAI-2 could inhibit efficiently the degradation of 3H-ECM by tumor cells (86.5%). The PAI-2 transfectants remained tumorigenic in nude mice, but tumors originating from the PAI-2 transfectants showed histologically the presence of a thick capsule which was absent in tumors from control cells.


[In vitro amplification of CD4(+) CD25(+) regulatory T cells and identification of amplified T cell immunosuppressive function].
This study was purposed to compare the effect of 3 different cell components for expanding CD4(+) CD25(+) Treg in vitro, and identify their immunosuppressive function. CD4(+) T cells, CD4(+) CD25(-)T cells and CD4(+) CD25(+)T cells were isolated from mouse splenocytes by MACS and then expanded in vitro. Phenotype of the T cell lines and expression of the FOXP3 was determined by flow cytometry. The inhibitory effect of expanded CD4(+) CD25(+) T cells on CD4(+) CD25(-)T cells was tested by MLR method. The results showed that the Treg cells from all the three groups were expanded significantly after culture for 2 weeks. In the CD4(+) T cells group, the proliferation rate was (77.8 ± 5.32) folds with a percentage of Treg cells increasing from (6.61 ± 1.00)% to (15.33 ± 1.31)%. The proliferation rate in the CD4(+) CD25(-) T cells group was (95.20 ± 7.67) folds, with the percentage of CD4(+) CD25(+) T cells raising from (0.37 ± 0.13)% to (9.84 ± 0.98)%. The proliferation rate in the CD4(+) CD25(+) T cells group was (41.20 ± 6.92) folds, the proportion of Treg cells decreased from (86.75 ± 1.25)% to (85.32 ± 1.62)%, and the expression of Foxp3 decreased from (76.92 ± 1.72)% to (75.33 ± 2.11)% during the culture, there were not significant differences in the cell purity and the expression of Foxp3, compared with pre-amplification. The inhibitory test showed that the expanded CD4(+) CD25(+) T cells could inhibit the proliferation of CD4(+) CD25(-) T cells in vitro in a cell dose-dependent manner. It is concluded that the amplification of CD4(+) CD25(+) Treg cells is successful in vitro, especially in the CD4(+) CD25(+) T cells group, the cell purity and Foxp3 gene is not obviously changes after amplification.


A Rational Design of α-Helix-Shaped Peptides Employing the Hydrogen-Bond Surrogate Approach: A Modulation Strategy for Ras-RasGRF1 Interaction in Neuropsychiatric Disorders.
In the last two decades, abnormal Ras (rat sarcoma protein)-ERK (extracellular signal-regulated kinase) signalling in the brain has been involved in a variety of neuropsychiatric disorders, including drug addiction, certain forms of intellectual disability, and autism spectrum disorder. Modulation of membrane-receptor-mediated Ras activation has been proposed as a potential target mechanism to attenuate ERK signalling in the brain. Previously, we showed that a cell penetrating peptide, RB3, was able to inhibit downstream signalling by preventing RasGRF1 (Ras guanine nucleotide-releasing factor 1), a neuronal specific GDP/GTP exchange factor, to bind Ras proteins, both in brain slices and in vivo, with an IC50 value in the micromolar range. The aim of this work was to mutate and improve this peptide through computer-aided techniques to increase its inhibitory activity against RasGRF1. The designed peptides were built based on the RB3 peptide structure corresponding to the α-helix of RasGRF1 responsible for Ras binding. For this purpose, the hydrogen-bond surrogate (HBS) approach was exploited to maintain the helical conformation of the designed peptides. Finally, residue scanning, MD simulations, and MM-GBSA calculations were used to identify 18 most promising α-helix-shaped peptides that will be assayed to check their potential activity against Ras-RasGRF1 and prevent downstream molecular events implicated in brain disorders.


The influence of Mycoplasma species on human and canine semen quality: a review.
Mycoplasma species (spp.) are bacteria that are difficult to detect. Currently, the polymerase chain reaction (PCR) is considered the most effective diagnostic tool to detect these microorganisms in both human and veterinary medicine. There are 13 known species of human Mycoplasma and 15 species of canine Mycoplasma. Owing to the difficulties in identifying the individual species of Mycoplasma, there is a lack of information regarding which species are saprophytic and which are pathogenic. The prevalence of the individual species is also unknown. In addition, in both humans and dogs, the results of some studies on the impact of Mycoplasma are conflicting. The presence of Mycoplasma spp. on the epithelium of reproductive tract is often associated with infertility, although they are also detected in healthy individuals. The occurrence of Mycoplasma spp. is more common in dogs (even 89%) than in humans (1.3%-4%). This is probably because the pH of a dog's genital is more conducive to the growth of Mycoplasma spp. than that of humans. Phylogenetically, human and canine Mycoplasma are related, and majority of them belong to the same taxonomic group. Furthermore, 40% of canine Mycoplasma spp. are placed in common clusters with those of human. This suggests that species from the same cluster can play a similar role in the canine and human reproductive tracts. This review summarizes the current state of knowledge about the impact of Mycoplasma on canine and human male fertility as well as the prospects of further development in this field.


Temporal outcomes of transcatheter mitral valve replacement in native mitral valve disease with annular calcification.
Surgical intervention in patients with native mitral disease due to severe mitral annular calcification (MAC) carries significant risk. Transcatheter mitral valve replacement (TMVR) using balloon-expandable aortic transcatheter heart valve (THV) in MAC had emerged as alternative treatment. We aim to study the temporal change in clinical outcomes of the procedure at a single center. We retrospectively studied 23 patients who underwent TMVR in MAC at Mayo Clinic from January, 14, 2014 to March, 15, 2019. Cases were divided into early (n = 11) and late (n = 12) experience. The primary end point was 30-day all-cause mortality. The secondary end points were immediate technical success, 30-day procedural success, and 1-year all-cause mortality. Mean age of patients was 75.2 ± 8.9 years and 17 (74.0%) were female. Median STS score for 30-day mortality was 8 (Interquartile range 4.3-13.4) for the entire population. Immediate technical success was achieved in 21 out of 23 patients (two failures in the early experience were related to tamponade and procedural death). Thirty-day procedural success was higher in the late experience (10 out of 12 patients) compared to early experience (5 out of 11 patients, p = .06). Four deaths in the first 30-days were observed in the early experience while all patients survived to hospital discharge in the late experience (p = .01). Procedural success and 30-day survival of transcatheter mitral valve replacement in severe mitral annular calcification procedure using balloon-expandable aortic prosthesis had improved over the years. This is likely attributed to significant advancement in procedural planning, valve design, and techniques.


A rapid and efficient method to generate multiple gene disruptions in Dictyostelium discoideum using a single selectable marker and the Cre-loxP system.
Dictyostelium discoideum has proven an exceptionally powerful system for studying numerous aspects of cellular and developmental functions. The relatively small ( approximately 34 Mb) chromosomal genome of Dictyostelium and high efficiency of targeted gene disruption have enabled researchers to characterize many specific gene functions. However, the number of selectable markers in Dictyostelium is restricted, as is the ability to perform effective genetic crosses between strains. Thus, it has been difficult to create multiple mutations within an individual cell to study epistatic relationships among genes or potential redundancies between various pathways. We now describe a robust system for the production of multiple gene mutations in Dictyostelium by recycling a single selectable marker, Blasticidin S resistance, using the Cre-loxP system. We confirm the effectiveness of the system by generating a single cell carrying four separate gene disruptions. Furthermore, the cells remain sensitive to transformation for additional targeted or random mutagenesis requiring Blasticidin selection and for functional expression studies of mutated or tagged proteins using other selectable markers.


Characterization of the rat oral microbiome and the effects of dietary nitrate.
The nitrate-nitrite-NO pathway to nitric oxide (NO) production is a symbiotic pathway in mammals that is dependent on nitrate reducing oral commensal bacteria. Studies suggest that by contributing NO to the mammalian host, the oral microbiome helps maintain cardiovascular health. To begin to understand how changes in oral microbiota affect physiological functions such as blood pressure, we have characterized the Wistar rat nitrate reducing oral microbiome. Using 16S rRNA gene sequencing and analysis we compare the native Wistar rat tongue microbiome to that of healthy humans and to that of rats with sodium nitrate and chlorhexidine mouthwash treatments. We demonstrate that the rat tongue microbiome is less diverse than the human tongue microbiome, but that the physiological activity is comparable, as sodium nitrate supplementation significantly lowered diastolic blood pressure in Wistar rats and also lowers blood pressure (diastolic and systolic) in humans. We also show for the first time that sodium nitrate supplementation alters the abundance of specific bacterial species on the tongue. Our results suggest that the changes in oral nitrate reducing bacteria may affect nitric oxide availability and physiological functions such as blood pressure. Understanding individual changes in human oral microbiome may offer novel dietary approaches to restore NO availability and blood pressure.


Usefulness of complete blood counts as a case-finding tool in medical outpatients.
To determine the usefulness of routine complete blood counts as a case-finding tool in medical outpatients. Prospective evaluation in a consecutive, unselected cohort. A university-based outpatient clinic providing primary and referral care in general internal medicine. A total of 595 patients (mean age [+/- SD], 40 +/- 15 years) making their initial clinic visit. A complete blood count was done in all patients. Components of the complete blood count that were not indicated for diagnostic or management purposes at the initial visit were classified as routine tests. The number and nature of new diagnoses leading to new patient management; the number of additional visits and diagnostic tests precipitated by positive results. Of the 2378 tests done, 1540 (65%) were routine tests. The percentages of abnormal test results for the four hematologic test components were as follows: leukocyte count, 7.6%; hemoglobin, 5.8%; mean corpuscular volume (MCV) of red blood cells, 7.9%; and platelet count, 4.5%. In only three patients (0.5%) was a new diagnosis made that led to a new management strategy (treatment of iron deficiency). Further work-up was unprofitable in 14 patients (2.4%). Two additional visits were necessary because of abnormal results on routine tests. No abnormality-associated clinical disease developed during a 15-month follow-up period in patients with unexplained abnormal results on routine hematologic tests. The routine complete blood count has limited usefulness as a case-finding tool and has a minimal effect on care in middle-aged medical outpatients.


Surface roughness measurement using white light speckle.
An experimental investigation is made of a technique for measurement of surface roughness. A speckle pattern is formed near the image plane of a lens in spatially coherent light having a broad spectral bandwidth. The contrast of the speckle is found to be related to surface roughness when the roughness and the coherence length of illumination are comparable in magnitude. Implementation techniques are discussed and results shown that indicate good correlation with profilometer measurements for a wide range of roughness values produced with a variety of finishing procedures.


Prayer Camps and Biomedical Care in Ghana: Is Collaboration in Mental Health Care Possible?
Experts have suggested that intersectoral partnerships between prayer camps and biomedical care providers may be an effective strategy to address the overwhelming shortage of mental health care workers in Africa and other low-income settings. Nevertheless, previous studies have not explored whether the prayer camp and biomedical staff beliefs and practices provide sufficient common ground to enable cooperative relationships. Therefore, we sought to examine the beliefs and practices of prayer camp staff and the perspective of biomedical care providers, with the goal of characterizing interest in-and potential for-intersectoral partnership between prayer camp staff and biomedical care providers. We conducted 50 open-ended, semi-structured interviews with prophets and staff at nine Christian prayer camps in Ghana, and with staff within Ghana's three public psychiatric hospitals. We used the purposive sampling method to recruit participants and the constant comparative method for qualitative data analysis. Prayer camp staff expressed interest in collaboration with biomedical mental health care providers, particularly if partnerships could provide technical support introducing medications in the prayer camp and address key shortcomings in their infrastructure and hygienic conditions. Nevertheless, challenges for collaboration were apparent as prayer camp staff expressed strong beliefs in a spiritual rather than biomedical explanatory model for mental illness, frequently used fasting and chained restraints in the course of treatment, and endorsed only short-term use of medication to treat mental illness-expressing concerns that long-term medication regimens masked underlying spiritual causes of illness. Biomedical providers were skeptical about the spiritual interpretations of mental illness held by faith healers, and were concerned by the use of chains, fasting, and the lack of adequate living facilities for patients in prayer camps; many, however, expressed interest in engaging with prayer camps to expand access to clinical care for patients residing in the camps. The findings demonstrate that biomedical care providers are interested in engaging with prayer camps. Key areas where partnerships may best improve conditions for patients at prayer camps include collaborating on creating safe and secure physical spaces and delivering medication for mental illness to patients living in prayer camps. However, while prayer camp staff are willing to engage biomedical knowledge, deeply held beliefs and routine practices of faith and biomedical healers are difficult to reconcile Additional discussion is needed to find the common ground on which the scarce resources for mental health care in Ghana can collaborate most effectively.


Polypeptide-catalyzed biosilicification of dentin surfaces.
In situ formation of mineral particles by biocatalysis would be advantageous for occluding dentin tubules to reduce permeability or for sealing of material-tooth interfaces. One approach would require that the peptide-catalyst remain functional on the dentin surface. Based on recent observations of retained activity on other surfaces, we hypothesized that poly(L-lysine) (PLL), an analog of the protein catalyst responsible for silica formation in primitive marine species, would remain functional on dentin. PLL was applied to dentin discs along with a pre-hydrolyzed silica precursor, tetramethyl orthosilicate (TMOS). Discs were analyzed microscopically (scanning electron microscopy, SEM) and chemically (x-ray photoelectron spectroscopy, XPS). The treated discs, but not the negative controls, exhibited partial distinct coating whose XPS survey was consistent with that of silica, demonstrating that the polypeptide was required and retained its mediating activity. Peptide-catalysts that mediate mineral formation can retain functionality on dentin, suggesting a wide range of preventive and treatment strategies.


Design of a monolithic Michelson interferometer for fringe imaging in a near-field, UV, direct-detection Doppler wind lidar.
The low-biased, fast, airborne, short-range, and range-resolved determination of atmospheric wind speeds plays a key role in wake vortex and turbulence mitigation strategies and would improve flight safety, comfort, and economy. In this work, a concept for an airborne, UV, direct-detection Doppler wind lidar receiver is presented. A monolithic, tilted, field-widened, fringe-imaging Michelson interferometer (FWFIMI) combines the advantages of low angular sensitivity, high thermo-mechanical stability, independence of the specific atmospheric conditions, and potential for fast data evaluation. Design and integration of the FWFIMI into a lidar receiver concept are described. Simulations help to evaluate the receiver design and prospect sufficient performance under different atmospheric conditions.


Rapid establishment of a COVID-19 perinatal biorepository: early lessons from the first 100 women enrolled.
Collection of biospecimens is a critical first step to understanding the impact of COVID-19 on pregnant women and newborns - vulnerable populations that are challenging to enroll and at risk of exclusion from research. We describe the establishment of a COVID-19 perinatal biorepository, the unique challenges imposed by the COVID-19 pandemic, and strategies used to overcome them. A transdisciplinary approach was developed to maximize the enrollment of pregnant women and their newborns into a COVID-19 prospective cohort and tissue biorepository, established on March 19, 2020 at Massachusetts General Hospital (MGH). The first SARS-CoV-2 positive pregnant woman was enrolled on April 2, and enrollment was expanded to SARS-CoV-2 negative controls on April 20. A unified enrollment strategy with a single consent process for pregnant women and newborns was implemented on May 4. SARS-CoV-2 status was determined by viral detection on RT-PCR of a nasopharyngeal swab. Wide-ranging and pregnancy-specific samples were collected from maternal participants during pregnancy and postpartum. Newborn samples were collected during the initial hospitalization. Between April 2 and June 9, 100 women and 78 newborns were enrolled in the MGH COVID-19 biorepository. The rate of dyad enrollment and number of samples collected per woman significantly increased after changes to enrollment strategy (from 5 to over 8 dyads/week, P < 0.0001, and from 7 to 9 samples, P < 0.01). The number of samples collected per woman was higher in SARS-CoV-2 negative than positive women (9 vs 7 samples, P = 0.0007). The highest sample yield was for placenta (96%), umbilical cord blood (93%), urine (99%), and maternal blood (91%). The lowest-yield sample types were maternal stool (30%) and breastmilk (22%). Of the 61 delivered women who also enrolled their newborns, fewer women agreed to neonatal blood compared to cord blood (39 vs 58, P < 0.0001). Establishing a COVID-19 perinatal biorepository required patient advocacy, transdisciplinary collaboration and creative solutions to unique challenges. This biorepository is unique in its comprehensive sample collection and the inclusion of a control population. It serves as an important resource for research into the impact of COVID-19 on pregnant women and newborns and provides lessons for future biorepository efforts.


Assessment of noncollinear spin-flip Tamm-Dancoff approximation time-dependent density-functional theory for the photochemical ring-opening of oxirane.
Under the usual assumption of noninteracting v-representability, density-functional theory (DFT) together with time-dependent DFT (TDDFT) provide a formally exact single-reference method suitable for the theoretical description of the electronic excited-states of large molecules, and hence for the description of excited-state potential energy surfaces important for photochemistry. The quality of this single-reference description is limited in practice by the need to use approximate exchange-correlation functionals. In particular it is far from clear how well approximations used in contemporary practical TDDFT calculations can describe funnel regions such as avoided crossings and conical intersections. These regions typically involve biradical-like structures associated with bond breaking and conventional wisdom would seem to suggest the need to introduce explicit double excitation character to describe these structures. Although this is lacking in ordinary spin-preserving (SP) TDDFT, it is present to some extent in spin-flip (SF) TDDFT. We report our tests of Wang-Ziegler noncollinear SF-TDDFT within the Tamm-Dancoff approximation for describing the avoided crossing in the C(2v) CC ring-opening reaction of oxirane and for describing the conical intersection relevant for the more physical asymmetric CO ring-opening reaction of oxirane. Comparisons are made with complete active space self-consistent field and quantum Monte Carlo benchmark results from two previous papers on the subject [J. Chem. Phys., 2007, 127, 164111; ibid 129, 2008, 124108]. While the avoided crossing in the C(2v) pathway is found to be reasonably well described, the method was found to be only partially successful for the conical intersection (CX) associated with the physically more important asymmetric pathway. The origin of the difficulties preventing the noncollinear SF-TDDFT method from giving a completely satisfactory description of the CX was traced back to the inability of SF-TDDFT based upon a single triplet reference state to correlate all potentially relevant configurations involving not just two but three nearly degenerate orbitals (n, σ(CO), and σ(CO)(*)). This article is also the first report of our implementation of SF-TDDFT within the deMon2k program.


The Assessment of Nipple Areola Complex Sensation with Semmes-Weinstein Monofilaments-Normative Values and Its Covariates.
To establish normative data for nipple-areola complex (NAC) sensibility examined with Semmes-Weinstein monofilament test (SWMT) and two-point discrimination (TPD) in women with varying breast sizes, including women with gigantomastia. We also aimed to identify clinical variables influencing NAC sensation. A total of 320 breasts in 160 Caucasian women (mean age 33.6 years, SD 11 years) were examined (including 50 hypertrophic breasts). NACs sensation was examined using Semmes-Weinstein monofilaments (SWM) and the Weber Two-Point Discrimination Test. The nipple appeared to be the most sensitive part of NAC. In normal-sized breasts, sensation thresholds (SWM) correlated with: age, BMI, history of births, breast size and ptosis (for all locations), breastfeeding history (for nipple and upper areola) and areola diameter (for all locations apart from the nipple). Regression analysis showed that age, cup size and suprasternal notch-to-nipple distance are risk factors for diminished NAC sensation. Sensation thresholds in all NAC locations of hypertrophic breasts were significantly higher compared to normal-sized breasts, while TPD tests did not differ between the groups. We provided normative values of NAC sensation (tactile threshold and TPD) for different NAC areas. Our investigation indicated that SWM are useful diagnostic tools when the following factors are considered while examining NAC sensation: location (nipple vs. areola), age, breast size, suprasternal notch-to-nipple distance, history of births and breastfeeding. Hypertrophic breasts presented significantly higher sensation thresholds for all NAC locations. The report may serve as a reference data for further investigations regarding NAC sensation after different breast surgeries.


DnaC, the indispensable companion of DnaB helicase, controls the accessibility of DnaB helicase by primase.
Former studies relying on hydrogen/deuterium exchange analysis suggest that DnaC bound to DnaB alters the conformation of the N-terminal domain (NTD) of DnaB to impair the ability of this DNA helicase to interact with primase. Supporting this idea, the work described herein based on biosensor experiments and enzyme-linked immunosorbent assays shows that the DnaB-DnaC complex binds poorly to primase in comparison with DnaB alone. Using a structural model of DnaB complexed with the C-terminal domain of primase, we found that Ile-85 is located at the interface in the NTD of DnaB that contacts primase. An alanine substitution for Ile-85 specifically interfered with this interaction and impeded DnaB function in DNA replication, but not its activity as a DNA helicase or its ability to bind to ssDNA. By comparison, substitutions of Asn for Ile-136 (I136N) and Thr for Ile-142 (I142T) in a subdomain previously named the helical hairpin in the NTD of DnaB altered the conformation of the helical hairpin and/or compromised its pairwise arrangement with the companion subdomain in each brace of protomers of the DnaB hexamer. In contrast with the I85A mutant, the latter were defective in DNA replication due to impaired binding to both ssDNA and primase. In view of these findings, we propose that DnaC controls the ability of DnaB to interact with primase by modifying the conformation of the NTD of DnaB.


Colocalization of galanin and prolactin within secretory granules of anterior pituitary cells in estrogen-treated Fischer 344 rats.
Galanin is localized within specific cell types of the rat anterior pituitary gland (AP). Immunocytochemical studies at the light microscope level have shown that lactotrophs, somatotrophs, and thyrotrophs contain galanin in the intact female rat, whereas lactotrophs in the male AP do not. We recently reported that galanin and PRL release from estrogen-treated male and female pituitary cells in culture are coregulated by dopamine, TRH, and somatostatin. This suggested that galanin is stored within secretory granules, conceivably with PRL. Using postembedding immunocytochemistry at the ultrastructural level, the objectives of this study were to: 1) determine the subcellular location of galanin in the AP; 2) elucidate if galanin and PRL are colocalized within the same secretory granules; and 3) compare the cellular localization of galanin in the male and female AP. Male and ovariectomized female (OVEX) Fischer 344 rats were implanted with estradiol-containing or empty Silastic capsules for 2 weeks. Postembedding immunogold labeling was performed using rabbit (for galanin) and guinea pig (for PRL) generated antisera. Two different sizes of colloidal gold spheres were used to localize the hormones in the same tissue section. Galanin was primarily localized in secretory granules of adenohypophyseal cells. Based upon immunocytochemical results and morphological criteria, galanin was contained in somatotrophs but not lactotrophs in the male and OVEX AP. The AP of estrogen-treated rats contained more specific immunogold labeling for galanin than untreated rats. The increased immunoreactivity for galanin was notably associated with lactotrophs. After exposure to estrogen, galanin and PRL were colocalized within the same secretory granules of the male and OVEX pituitary cells. We conclude: 1) galanin is localized within secretory granules of the rat AP; 2) galanin and PRL are colocalized within secretory granules of the male and OVEX AP after estrogen treatment; and 3) galanin is localized in similar cell types in the male and OVEX AP, before and after estrogen treatment. These data provide a morphological basis for the coregulation of galanin and PRL secretion by hypothalamic factors.


Utility of biochemical markers and RVD/LVD ratio in acute pulmonary embolism risk classification in Emergency Department.
We aimed to determine the efficacy of troponin I, D-dimer, and lactate levels and right ventricular diameter (RVD)/left ventricular diameter (LVD) ratio on pulmonary computed tomography angiography (PCTA) in the risk classification of patients who were diagnosed with acute pulmonary embolism (APE) in Emergency Department (ED). Patients who were diagnosed as having APE by PCTA in ED were included in this retrospective study. Patients were grouped as high-risk (undergoing cardiopulmonary resuscitation or given thrombolytic therapy), moderate-risk (with non-high-risk and positive ECO findings) and low risk (others). Troponin I, D-dimer, and lactate levels of patients were determined. RVD, RVD/LVD ratio, and interventricular septum deviation were calculated from PCTA images. A total of 121 patients were included (35 high, 36 moderate, 50 low risk). Lactate was different in the high-risk group from the other groups, whereas there was no difference between the moderate and low-risk groups. Troponin I levels were not different between the high-risk and moderate-risk groups. There were statistically significant differences between the high, moderate, and low-risk groups in terms of mean RVD/LVD ratios. ROC analyses performed in order to define high-risk group revealed a cut-off value of > 2.3 (AUC = 0.848, sensitivity = 70%, specificity = 90%, + Likelihood ratio (LR) = 7, -LR = 0.33, 95% CI = 0.752-0.943) for lactate and > 1.40 (AUC = 0.695 sensitivity = 71%, specificity = 80%, + LR = 3.6, -LR = 0.36, 95% CI = 0.668-0.822) for RVD/LVD ratio. Lactate levels and RVD/LVD ratio were shown to be useful in distinguishing high-risk patients from other patient groups. Troponin I is important in terms of showing cardiac involvement, but it is inadequate in distinguishing between high and moderate-risk patients. Lactate, troponin I, and RVD/LVD ratio may be used together for a more accurate separation of patients with high, intermediate and low-risk.


Mechanisms underlying lung resistance-related protein (LRP)-mediated doxorubicin resistance of non-small cell lung cancer cells.
Lung resistance-related protein (LRP) is a human major vault protein (MVP) implicated in drug resistance of cancer cells in a cell-type dependent manner. The primary goal of the study was to understand the role(s) of LRP in doxorubicin (DOX) resistance of non-small cell lung cancer (NSCLC) cells and the underlying working mechanisms. In the study, the roles of LRP in the regulation of DOX dynamics, nuclear import of minor vault proteins (vault poly (ADP-ribose) polymerase, vPARP and telomerase associated protein-1, TEP-1) and DOX-mediated cytotoxicity were examined in CH27 and H460 cells. Our results were the first to show that the CH27 cells with higher LRP expression levels were more resistant to DOX-induced cytotoxicity as shown in apoptosis experiments. LRP at the nuclear membrane could regulate DOX efflux from the nucleus to the cytosol, and also the reverse vPARP/TEP1 influx from the cytosol, to protect NSCLC cells from DOX-induced apoptosis. Cytosolic LRP could bind to DOX, vPARP and TEP1 to clear DOX away from the nucleus and promote the assembly of vaults for cell protection again. Based on the data obtained, the molecular mechanisms responsible for DOX resistance of NSCLC were delineated to demonstrate that LRP, vPARP and TEP1 were potential targets for NSCLC therapy. Inhibitors of these proteins, including small interfering LRP (siLRP), wheat-germ agglutenin (WGA) (WGA), 3-aminobenzamide (3-AB) and 3,6,9-trisubstituted acridine 9-[4-(N,N-dimethylamino) phenylamino]-3,6-bis(3-pyrrolodinopropionamido) acridine (BRACO-19), break down the DOX resistance of NSCLC cells, particularly in CH27 cells, and may have therapeutic values in the control of NSCLC.


The Use of Sour and Sweet Whey in Producing Compositions with Pleasant Aromas Using the Mold Galactomyces geotrichum: Identification of Key Odorants.
Fermented products with a pleasant aroma and with strong honey, rose, and fruit odor notes were developed through the biotransformation of a medium containing sour or sweet whey with the addition of l-phenylalanine by the Galactomyces geotrichum mold. In order to obtain the strong honey-rose aroma, G. geotrichum strains were screened and fermentation conditions were optimized to achieve a preferable ratio (>1) of phenylacetaldehyde to 2-phenylethanol by the Ehrlich pathway. This allowed post-fermentation products with the ratio of concentrations of phenylacetaldehyde to 2-phenylethanol being 1.7:1. Additionally, the use of gas chromatography-olfactometry (GC-O) analysis and the calculation of odor activity values (OAVs) allowed 10 key odorants to be identified in post-fermentation products. The highest OAVs were found for phenylacetaldehyde with a honey odor in both sour and sweet whey cultures (3010 and 1776, respectively). In the variant with sour whey, the following compounds with the highest OAVs were 3-methyl-1-butanol (131), 3-(methylthio)-propanal (119), 3-methylbutanal (90), dimethyl trisulfide (71), 2,3-butanedione (37), and 2-phenylethanol (29). In the post-fermentation product with sweet whey, the following compounds with the highest OAVs were 3-(methylthio)-propanal (112), dimethyl trisulfide (69), and 2,3-butanedione (41).


The effects of calcium on the thermal stability and activity of manganese peroxidase.
The presence of micromolar Ca2+ efficiently prevented the thermal inactivation of manganese peroxidase from Phanerochaete chrysosporium. The amount of Ca2+ normally present in the enzyme decreased when the enzyme was thermally inactivated and EGTA increased the rate of inactivation. The inactivation kinetics were biphasic, suggesting a sequential two-step process. The rate of inactivation during the second, slower step corresponded to the rate of loss of heme from the enzyme. Thermally inactivated manganese peroxidase could be readily reactivated in the presence of excess Ca2+. However, as the time of thermal incubation increased and the amount of remaining heme decreased, the amount of enzyme activity recovered decreased. Therefore, while both steps of denaturation could be prevented by Ca2+, only one step could be reversed upon the addition of Ca2+. It is proposed that the first step of denaturation involves the loss of Ca2+ which causes conformational changes resulting in the loss of manganese peroxidase activity. The second step is believed to involve further structural loss and results in the loss of heme from the enzyme. It is concluded that manganese peroxidase is susceptible to thermal inactivation because it contains relatively labile Ca2+ ions required for stability and activity.


Divided attention and the encoding effects of retrieval.
Retrieving from memory both reveals as well as modifies memory. It is important to understand how these encoding effects of retrieval differ from other forms of encoding. One possible difference relates to attention: divided attention is well known to disrupt memory encoding but typically has much less impact on memory retrieval. However, less is known about the relative attentional demands of the encoding consequences of retrieval. The current experiments examined retrieval-based encoding using free recall, a retrieval task purported to require substantial attentional resources. In three experiments, participants studied common category exemplars (Phase 1), restudied or freely recalled the exemplars (Phase 2), and then took a final free-recall test (Phase 3). Phase 2 occurred under full attention (FA) or divided attention (DA). In all three experiments, the negative effect of DA on final recall was significant in the restudy but not retrieval condition. The pattern persisted with short (Experiment 1) or long study lists (Experiment 2), requiring lesser or greater retrieval effort, and with multiple Phase 2 tests, permitting the development of more elaborate retrieval strategies (Experiment 3). The encoding effects of retrieval appear resilient to distraction, even using a memory task that is more effortful and easily disrupted by DA (i.e., free recall). In addition, these results are inconsistent with elaboration and effort accounts of retrieval-based learning.


[Expression of RASSF1A mRNA and its relationship with HPV-16 in cervical cancer].
To study the expression of RASSF1A mRNA and its relationship with HPV-16 in cervical cancer. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of RASSF1A mRNA in 56 patients with cervical cancer, 36 patients with CIN and 20 patients with normal cervix. HPV-16 DNA was also examined by PCR in 56 patients with cervical cancer. The rates of loss of RASSF1A mRNA expression were 0% (0/20), 22.2% (8/36) and 50.0% (28/56) in patients with normal cervix, CIN and cervical cancer respectively. The loss rate of RASSF1A mRNA in the cervical tissues at the advanced stages (stages III-IV, 82.4%) was higher than that at the early stages (stages I-II, 35.9%, P < 0.05). RASSF1A mRNA deletion occurred more often in the tissues of squamous cervical cancers (63.3%) than in those of adenocarcinoma (34.6%) (P < 0.05). The loss rate of the poor differentiation group was 70.8%, significantly higher than that of the middle and high differentiation group (37.5%, P < 0.05). The loss of RASSF1A was negatively related with HPV-16 infection in cervical cancers, especially in adenocarcinoma (P < 0.905). Loss of RASSF1A may play an important role in tumorgenesis and development of cervical cancer.


Making a medical practice in an uneasy world: some thoughts from a century ago.
A century ago medical trainees in the United States faced many of the same sorts of issues that confront trainees today. For practical advice in setting up their practices, many of those trainees turned to a book by physician David Cathell called The Physician Himself, which was first published in 1882 and was reprinted many times thereafter. This paper explores the advice Cathell offered in his book as well as parallels with events of the recent past, discussing Cathell's opinions on physician overcrowding, medical education, new technology such as the telephone and the automobile, and the sensitive ethical issues that physicians must deal with. The paper concludes that because being an excellent physician in both Cathell's time and today requires more than good science, Cathell's book helps to remind us of the social world in which medicine is practiced.


Z1456467176 alleviates gouty arthritis by allosterically modulating P2X7R to inhibit NLRP3 inflammasome activation.
NLRP3 inflammasome activation is a central process in initiating gout flares. The unique conformational rearrangement of the P2X7 receptor (P2X7R) upon ATP binding is critical for the activation of the NLRP3 inflammasome. However, studies on allosteric modulation of P2X7R in gout treatment are limited. Here, we aimed to investigate the therapeutic implications of targeting P2X7R in gout by designing a P2X7R allosteric inhibitor and validating the inhibitory function on NLRP3 inflammasome activation. Through virtual screening, we identified Z1456467176 (N-{3-[(2-aminoethyl) sulfamoyl] phenyl}-2-methyl-3-[3-(trifluoromethyl) phenyl] propanamide hydrochloride) bound to the drug-binding pocket as a potential antagonist of P2X7R. In functional assays, ATP- or BzATP-induced P2X7R function was assessed in vitro in HEK-293T cells overexpressing hP2X7R (dye uptake assay) and macrophages (IL-1β release assay). Z1456467176 exhibited a stable and significant P2X7R inhibitory effect. Importantly, in MSU crystal-induced gout, the presence and involvement of ATP were confirmed. Z1456467176 blocked ATP-induced activation of the NLRP3-caspase-1-IL-1β pathway and exerted promising effects in reducing gouty joint inflammation in rats. In addition, molecular docking and molecular dynamics simulation studies showed that the P27XR protein conformation was remodeled by Z1456467176 binding. Collectively, our results provide a potent P2X7R allosteric inhibitor that facilitates the remission of MSU crystal-induced gout inflammation by inhibiting NLRP3 inflammasome activation, suggesting that allosteric inhibition of P2X7R represents a new direction in gout treatment.


An atomic mutation cycle for exploring RNA's 2'-hydroxyl group.
The 2'-hydroxyl group fulfills numerous structural and functional roles in RNA, including those of hydrogen bond donor and acceptor. While loss of function upon 2'-deoxynucleotide substitution establishes the importance of specific 2'-hydroxyl groups within RNA, this approach provides no information about how these hydroxyl groups impart their functional contribution. We use an atomic mutation cycle to evaluate the functional importance of the 2'-hydroxyl group's hydrogen atom. Using the Tetrahymena ribozyme reaction, we challenge the cycle to expose the catalytic contribution of the cleavage site 2'-hydroxyl group and its associated hydrogen bond network. The results establish the viability of this cycle as an approach to reveal 2'-hydroxyl groups that donate functionally significant hydrogen bonds.


Thyroid growth stimulating immunoglobulins in sporadic and endemic colloid goitre.
Several methods have been described to measure immunoglobulins stimulating the growth of thyroid cells in vitro, the so called Thyroid Growth stimulating Immunoglobulins (TGI). The methods make use of either Ig-stimulated guinea pig thyroid (organ) cultures (the cytochemical bioassays; growth is measured via Feulgen-densitometry or pentose shunt analysis), the culture of Ig-stimulated rat or porcine thyroid follicles (growth is measured via 3H-thymidine incorporation), or of Ig-stimulated rat FRTL-5 cells (growth is measured via 3H-thymidine incorporation, or counting the number of mitoses in arrest). The various methods have now been validated: (a) the data obtained with TGI preparations in the Feulgen Cytochemical Bioassay (CBA's) correlate well with those obtained in the FRTL-5 mitosis arrest assay with the same TGI preparations, (b) the growth stimulation can not be ascribed to hormonal contaminations of the used Ig preparations, since protein A-sepharose purified IgG is active in the assays and since anti human IgG's neutralize the growth stimulating effects of the preparations. Using the assays TGI has been found positive in goitrous Graves patients, in sporadic goitre patients and in endemic goitre patients. Particularly, patients complaning of recurrent goitre after thyroidectomy or with recent goitre growth are TGI positive. TGI occurs in sporadic goitre in the absence of TSH-receptor antibodies. The autoimmune-prone animal model of the BB rat also proved to be TGI-positive; the animal shows an increased thyroid glandular weight. Both the histomorphology of human goitres as well as the goitre of the BB rat indicate that the dendritic cell plays a prominent role in initiating the thyroid autoimmune reaction.


Lack of adducin impairs the stability of endothelial adherens and tight junctions and may be required for cAMP-Rac1-mediated endothelial barrier stabilization.
Adducin (Add) is an actin binding protein participating in the stabilization of actin/spectrin networks, epithelial junctional turnover and cardiovascular disorders such as hypertension. Recently, we demonstrated that Add is required for adherens junctions (AJ) integrity. Here we hypothesized that Add regulates tight junctions (TJ) as well and may play a role in cAMP-mediated barrier enhancement. We evaluated the role of Add in MyEnd cells isolated from WT and Add-Knock-Out (KO) mice. Our results indicate that the lack of Add drastically alters the junctional localization and protein levels of major AJ and TJ components, including VE-Cadherin and claudin-5. We also showed that cAMP signaling induced by treatment with forskolin and rolipram (F/R) enhances the barrier integrity of WT but not Add-KO cells. The latter showed no junctional reorganization upon cAMP increase. The absence of Add also led to higher protein levels of the small GTPases Rac1 and RhoA. In vehicle-treated cells the activation level of Rac1 did not differ significantly when WT and Add-KO cells were compared. However, the lack of Add led to increased activity of RhoA. Moreover, F/R treatment triggered Rac1 activation only in WT cells. The function of Rac1 and RhoA per se was unaffected by the total ablation of Add, since direct activation with CN04 was still possible in both cell lines and led to improved endothelial barrier function. In the current study, we demonstrate that Add is required for the maintenance of endothelial barrier by regulating both AJ and TJ. Our data show that Add may act upstream of Rac1 as it is necessary for its activation via cAMP.


Three extracellular proteases from Cochliobolus carbonum: cloning and targeted disruption of ALP1.
Three extracellular serine proteases (Alp1a, Alp1b, Alp2) from Cochliobolus carbonum were purified and characterized. Of eight carbon/protein substrates tested, total protease activity was highest when the fungus was grown on medium containing collagen. Alp1a and Alp1b are members of the trypsin family (EC 126.96.36.199), and Alp2 is a member of the subtilisin family (EC 184.108.40.206). Alp1a, Alp1b, and Alp2 have monomer molecular masses of 25, 30, and 38 kDa respectively. Alp1b is glycosylated, whereas Alp1a is not. The gene encoding Alp1a, ALP1, was isolated using PCR primers based on two amino acid sequences: One obtained directly from the N-terminus of Alp1a and another that is highly conserved in other trypsins. The transcriptional start site was determined using RACE and the intron structure and polyadenylation site were determined from a cDNA clone. An internal fragment of ALP1 was used to create Alp1a null mutants by transformation-mediated gene disruption. Total protease activity in the mutants was reduced by 35% to 45%. By chromatographic analysis, the mutants had lost two peaks of UV absorption and the two protease activities corresponding to Alp1a and Alp1b, which, together with the biochemical data, indicates that Alp1a and Alp1b are products of the same gene. The in vitro growth and diseases phenotypes of the ALP1 mutants were distinguishable from the wild-type strain; therefore, ALP1 is not by itself required for pathogenicity.


Comparing the Impact of Differing Preschool Autism Interventions on Parents in Two Canadian Provinces.
Early Intensive Behavioural Intervention (EIBI) is effective for preschoolers with autism spectrum disorder (ASD). Parental measures are rarely included in EIBI effectiveness studies, yet parental distress and lower self-efficacy are associated with poorer child outcomes. Parents of preschoolers with ASD (N = 485) were surveyed at baseline (T1), one-year post-intervention (T2), and school entry (T3) about family distress/crisis, parental self-efficacy, and satisfaction with services in two Canadian provinces. Family distress/crisis decreased and parental self-efficacy increased from T1 to T2. Increases in self-efficacy were largely maintained at T3. Parents were highly satisfied with services. Greater satisfaction for those residing in the province utilizing a parent-coaching model suggests that parent involvement is associated with positive parent outcomes.


[Dysphagia and cervical osteophytosis: case report and review of the literature].
Dysphagia with anterior osteophytosis of the cervical bone is a more and more reported affection, especially after the age of 60. However it remains a rare event. The cause to effect relationship between these two entities should be documented by a complete assessment. Surgical treatment is reserved for patients whose medical treatment has failed. We describe an original case of cervical bone osteophytosis which was diagnosed during an acute and febrile aphagia and review the literature about its assessment and treatment.


Software license agreements: don't get locked into bad deals. Do's and don'ts of negotiating the best long-term arrangement with IT vendors.
Selecting, implementing and operating the right technology solutions are increasingly critical components for health plans in today's competitive consumer health care market. Decisions about what architecture to employ, what software applications to buy and what hardware platform to run affect a plan's ability to become a market leader. Additionally, health plans must consider the best approaches for maintaining and operating their IT solutions, which inevitably include employing a global sourcing strategy that extends IT budgets and expertise while lowering costs.


Skull parosteal lipoma with reactive hyperostosis: a case report.
A 50-year-old female presented with more than 20-year history of a large subcutaneous mass in the left parieto-occipital portion. Magnetic resonance (MR) imaging revealed the lipomatous mass to show a high signal intensity in both T1- and T2-weighted images. A part of the lipomatous lesion progressed into the underlying hyperostosis and skull. The preoperative diagnosis was skull invasion of a well-differentiated liposarcoma. The tumor was removed completely, including the underlying hyperostosis and skull. Microscopy confirmed a lipoma without any lipoblasts, which was firmly attached to the reactive hyperostosis, and islands of lipoma were involved in the underlying hyperostosis and skull cortex. A pathological diagnosis of parosteal lipoma with reactive hyperostosis was made. Long-term progression of parosteal lipoma may cause to involve the underlying hyperostosis and skull, and led to the diagnosis of invasion of a malignant tumor on MR imaging.


[Specific features of nitrogen transformation in the soddy-podzolic soil in the areas of habitat of the common vole Microtus arvalis].
The influence of common vole Microtus arvalis on processes of nitrogen fixation and denitrification in the soddy-podsolic soil was studied. In the common vole colonies, the level of nitrogen fixation was reliably lower and that of denitrification higher, than in the control soil outside these colonies. Nitrogen-containing excretory products of voles accumulating in the soil are among the main factors that determine the activity of these processes.


Plasma diamine oxidase activity is greater in copper-adequate than copper-marginal or copper-deficient rats.
The object of this study was to determine whether serum diamine oxidase activity could distinguish among adequate, marginal and deficient copper status in rats. Male weanling Sprague-Dawley rats (n = 21) were randomly assigned to one of three dietary regimens, with copper concentrations of 0.52, 1.73 and 6.7 mg/kg diet. On completion of the study, body weights were significantly different among dietary groups, with copper-marginal rats displaying the highest mean weight and copper-deficient rats the lowest. Copper-deficient rats ate significantly less food than the other two groups. Rats fed the three diets had significantly different liver copper concentrations. Liver and heart superoxide dismutase and cytochrome c oxidase activities, and plasma ceruloplasmin and erythrocyte superoxide dismutase activities were significantly lower in the copper-deficient rats than in the other two groups. Plasma diamine oxidase activity was lower in both copper-deficient (0.18 +/- 0.11 U/L) and marginal (0.21 +/- 0.11 U/L) rats compared with copper-adequate rats (3.35 +/- 0.28 U/L). Of the biochemical indices measured, only liver copper concentration (-20%) and plasma diamine oxidase activity (-94%) differed between rats fed copper-marginal and copper-adequate diets. Plasma diamine oxidase activity, therefore, may be a sensitive functional biomarker of suboptimal copper status.


Differential usage of multiple brain-derived neurotrophic factor promoters in the rat brain following neuronal activation.
The rat brain-derived neurotropic factor (BDNF) gene consists of four 5' exons linked to separate promoters and one 3' exon encoding the prepro-BDNF protein. To gain insights into the regulation of BDNF mRNA expression, probes specific for the different 5' exons were used to study the expression of BDNF mRNA in the brain. Following a systemic injection of the glutamate analog kainic acid, exon I, II, and III mRNAs increased transiently in hippocampus and cerebral cortex. A modest increase was seen for exon IV, where a new transcription initiation site was induced by this treatment. Pretreatments with the N-methyl-D-aspartate (NMDA) receptor antagonist MK801 or the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antagonist 2,3-dihydroxy-6-nitrosulfanoylbenzo(f)quinoxaline revealed two region-specific patterns of glutamate receptor-mediated regulation. The first pattern found in neocortex, piriform cortex, and amygdala involves regulation of BDNF exon I, II, and III mRNAs through NMDA and AMPA/kainate receptors. The second pattern found in the hippocampus involves regulation of BDNF exon I, II, and III mRNAs by high-affinity kainate or metabotropic receptors. Treatment with the gamma-aminobutyric acid subtype A (GABAA) receptor antagonist bicuculline increased exon I and III mRNAs in the denate gyrus, and the muscarinic receptor agonist pilocarpine increased exon I mRNA mainly in the neocortex. These data show that the four BDNF promoters allow multiple points of BDNF mRNA regulation and suggest that the activation of different subtypes of glutamate receptors differentially regulates the expression of BDNF exon-specific mRNAs in the brain.