Stringent and reproducible tetracycline-regulated transgene expression by site-specific insertion at chromosomal loci with pre-characterised induction characteristics.
The ability to regulate transgene expression has many applications, mostly concerning the analysis of gene function. Desirable induction characteristics, such as low un-induced expression, high induced expression and limited cellular heterogeneity, can be seriously impaired by chromosomal position effects at the site of transgene integration. Many clones may therefore need to be screened before one with optimal induction characteristics is identified. Furthermore, such screens must be repeated for each new transgene investigated, and comparisons between clones with different transgenes is complicated by their different integration sites. To circumvent these problems we have developed a "screen and insert" strategy in which clones carrying a transgene for a fluorescent reporter are first screened for those with optimal induction characteristics. Site-specific recombination (SSR) is then be used repeatedly to insert any new transgene at the reporter transgene locus of such clones so that optimal induction characteristics are conferred upon it. Here we have tested in a human fibrosarcoma cell line (HT1080) two of many possible implementations of this approach. Clones (e.g. Rht14-10) in which a GFP reporter gene is very stringently regulated by the tetracycline (tet) transactivator (tTA) protein were first identified flow-cytometrically. Transgenes encoding luciferase, I-SceI endonuclease or Rad52 were then inserted by SSR at a LoxP site adjacent to the GFP gene resulting stringent tet-regulated transgene expression. In clone Rht14-10, increases in expression from essentially background levels (+tet) to more than 104-fold above background (-tet) were reproducibly detected after Cre-mediated insertion of either the luciferase or the I-SceI transgenes. Although previous methods have made use of SSR to integrate transgenes at defined sites, none has effectively combined this with a pre-selection step to identify integration sites that support optimal regulatory characteristics. Rht14-10 and similar HT1080-derived clones can now be used in conjunction with a convenient delivery vector (pIN2-neoMCS), in a simple 3-step protocol leading to stringent and reproducible transgene regulation. This approach will be particularly useful for transgenes whose products are very active at low concentrations and/or for comparisons of multiple related transgenes.


Discovery of Infection Associated Metabolic Markers in Human African Trypanosomiasis.
Human African trypanosomiasis (HAT) remains a major neglected tropical disease in Sub-Saharan Africa. As clinical symptoms are usually non-specific, new diagnostic and prognostic markers are urgently needed to enhance the number of identified cases and optimise treatment. This is particularly important for disease caused by Trypanosoma brucei rhodesiense, where indirect immunodiagnostic approaches have to date been unsuccessful. We have conducted global metabolic profiling of plasma from T.b.rhodesiense HAT patients and endemic controls, using 1H nuclear magnetic resonance (NMR) spectroscopy and ultra-performance liquid chromatography, coupled with mass spectrometry (UPLC-MS) and identified differences in the lipid, amino acid and metabolite profiles. Altogether 16 significantly disease discriminatory metabolite markers were found using NMR, and a further 37 lipid markers via UPLC-MS. These included significantly higher levels of phenylalanine, formate, creatinine, N-acetylated glycoprotein and triglycerides in patients relative to controls. HAT patients also displayed lower concentrations of histidine, sphingomyelins, lysophosphatidylcholines, and several polyunsaturated phosphatidylcholines. While the disease metabolite profile was partially consistent with previous data published in experimental rodent infection, we also found unique lipid and amino acid profile markers highlighting subtle but important differences between the host response to trypanosome infections between animal models and natural human infections. Our results demonstrate the potential of metabolic profiling in the identification of novel diagnostic biomarkers and the elucidation of pathogenetic mechanisms in this disease.


Baseline chest radiograph for lung cancer detection in the randomized Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial.
The Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial was initiated in 1992 to examine cause-specific mortality reduction from screening for these four cancers in men and women. We report lung cancer detection results of the baseline screening round. Of the 154,942 participants enrolled, who were aged 55-74 years with no history of PLCO cancers, 77,465 were randomly assigned to the intervention arm. Current or former smokers and never smokers in this arm received an initial single-view posterior-anterior chest radiograph. In the initial screen, 5991 (8.9%, 95% confidence interval [CI] = 8.7% to 9.2%) of radiographs were suspicious for lung cancer: 8.2% (95% CI = 7.9% to 8.5%) for women and 9.6% (95% CI = 9.3% to 10.0%) for men. Rates were highest for older age groups and for smokers. Among those 5991 participants with a positive screen, 206 (3.4%, 95% CI = 3.0% to 3.9%) underwent biopsy examination, 126 (61.2%, 95% CI = 54.5% to 67.8%) of whom were diagnosed with lung cancer within 12 months of the screen (59 in women and 67 in men). The positive predictive value was 2.1% (95% CI = 1.7% to 2.5%), and 1.9 lung cancers were detected per 1000 screens. Among these cancers, 44% (95% CI = 35% to 52%) were stage I non-small-cell lung cancer. High rates of lung cancer were found in current smokers (6.3 per 1000 screens) and in former smokers who had smoked within the past 15 years (4.9 per 1000 screens). The lung cancer detection rate among never smokers was 0.4 per 1000 screens; this group accounted for 11% (95% CI = 5.6% to 16.6%) of the cancers identified. In the baseline screen, nearly half the cancers were stage I. Whether this experience results in a reduction in lung cancer mortality is yet to be seen.


Ursolic Acid Attenuates Hepatic Steatosis, Fibrosis, and Insulin Resistance by Modulating the Circadian Rhythm Pathway in Diet-Induced Obese Mice.
The aim of the current study was to elucidate the effects of long-term supplementation with dietary ursolic acid (UR) on obesity and associated comorbidities by analyzing transcriptional and metabolic responses, focusing on the role of UR in the modulation of the circadian rhythm pathway in particular. C57BL/6J mice were divided into three groups and fed a normal diet, high-fat diet, or high-fat + 0.05% (w/w) UR diet for 16 weeks. Oligonucleotide microarray profiling revealed that UR is an effective regulator of the liver transcriptome, and canonical pathways associated with the "circadian rhythm" and "extracellular matrix (ECM)⁻receptor interactions" were effectively regulated by UR in the liver. UR altered the expression of various clock and clock-controlled genes (CCGs), which may be linked to the improvement of hepatic steatosis and fibrosis via lipid metabolism control and detoxification enhancement. UR reduced excessive reactive oxygen species production, adipokine/cytokine dysregulation, and ECM accumulation in the liver, which also contributed to improve hepatic lipotoxicity and fibrosis. Moreover, UR improved pancreatic islet dysfunction, and suppressed hepatic gluconeogenesis, thereby reducing obesity-associated insulin resistance. Therapeutic approaches targeting hepatic circadian clock and CCGs using UR may ameliorate the deleterious effects of diet-induced obesity and associated complications such as hepatic fibrosis.


The crystal structure of urease from Klebsiella aerogenes.
The crystal structure of urease from Klebsiella aerogenes has been determined at 2.2 A resolution and refined to an R factor of 18.2 percent. The enzyme contains four structural domains: three with novel folds playing structural roles, and an (alpha beta)8 barrel domain, which contains the bi-nickel center. The two active site nickels are 3.5 A apart. One nickel ion is coordinated by three ligands (with low occupancy of a fourth ligand) and the second is coordinated by five ligands. A carbamylated lysine provides an oxygen ligand to each nickel, explaining why carbon dioxide is required for the activation of urease apoenzyme. The structure is compatible with a catalytic mechanism whereby urea ligates Ni-1 to complete its tetrahedral coordination and a hydroxide ligand of Ni-2 attacks the carbonyl carbon. A surprisingly high structural similarity between the urease catalytic domain and that of the zinc-dependent adenosine deaminase reveals a remarkable example of active site divergence.


A new method for estimating left ventricular dP/dt by continuous wave Doppler-echocardiography. Validation studies at cardiac catheterization.
In this study, we explored the use of continuous wave Doppler-echocardiography guided by color Doppler flow-mapping as a method for noninvasively calculating the rate of pressure rise (RPR) in the left ventricle. Continuous wave Doppler determination of the velocities in mitral regurgitant jets allows calculation of instantaneous pressure gradients between the left ventricle and the left atrium. Left atrial pressure variations in early systole can be considered negligible; therefore, the rising segment of the mitral regurgitation velocity curve should reflect left ventricular pressure increase. We studied 50 patients (mean age, 51 years; range, 25-66 years) in normal sinus rhythm with color Doppler-proven mitral regurgitation and compared the Doppler-derived left ventricular RPR with peak dP/dt obtained at cardiac catheterization. Doppler studies were performed simultaneously with cardiac catheterization in 11 patients and immediately before in the remaining cases. Two points were arbitrarily selected on the steepest rising segment of the continuous wave mitral regurgitation velocity curve (point A, 1 m/sec, point B, 3 m/sec), and the time interval (t) between them was measured. Following the Bernoulli relation, the pressure rise between points A and B is 32 mm Hg (4vB2-4vA2) and the RPR is 32 mm Hg/t. Results showed a linear correlation between the Doppler RPR and peak dP/dt (r = 0.87, SEE = 316 mm Hg/sec). The RPR in the left ventricle can be derived from the continuous wave Doppler mitral regurgitation velocity curve.


Roles of V antigen in promoting virulence and immunity in yersiniae.
It is established that yersiniae harboring an approximately 45-megadalton Vwa-plasmid can produce V and W antigens (Vwa+), and that sera containing anti-V provides passive protection to mice against Yersinia pestis. This observation was extended by the use of monospecific anti-V prepared by injecting rabbits with partially purified V, absorption of antisera with a Vwa- extract, and then separation of gamma-globulin by traditional processes of fractionation or by affinity chromatography. These preparations provided passive protection against 10 minimum lethal doses of virulent Y. pestis KIM, Yersinia pseudotuberculosis PB1, and Yersinia enterocolitica WA. Kinetics of elimination of these Vwa+ yersiniae from organs and blood of passively immunized mice closely resembled those of avirulent Vwa- mutants from normal mice. Injection into mice of sterile crude extracts of Y. pseudotuberculosis PB1 containing V promoted significant survival and retention of Vwa- mutants of Y. pestis, Y. pseudotuberculosis, and Y. enterocolitica. This effect was eliminated by the removal of V before injection by precipitation with monospecific antibody. These results indicate that V antigen per se is the major virulence factor mediated by Vwa-plasmids.


Proposing an empirically justified reference threshold for blood culture sampling rates in intensive care units.
Early and appropriate blood culture sampling is recommended as a standard of care for patients with suspected bloodstream infections (BSI) but is rarely taken into account when quality indicators for BSI are evaluated. To date, sampling of about 100 to 200 blood culture sets per 1,000 patient-days is recommended as the target range for blood culture rates. However, the empirical basis of this recommendation is not clear. The aim of the current study was to analyze the association between blood culture rates and observed BSI rates and to derive a reference threshold for blood culture rates in intensive care units (ICUs). This study is based on data from 223 ICUs taking part in the German hospital infection surveillance system. We applied locally weighted regression and segmented Poisson regression to assess the association between blood culture rates and BSI rates. Below 80 to 90 blood culture sets per 1,000 patient-days, observed BSI rates increased with increasing blood culture rates, while there was no further increase above this threshold. Segmented Poisson regression located the threshold at 87 (95% confidence interval, 54 to 120) blood culture sets per 1,000 patient-days. Only one-third of the investigated ICUs displayed blood culture rates above this threshold. We provided empirical justification for a blood culture target threshold in ICUs. In the majority of the studied ICUs, blood culture sampling rates were below this threshold. This suggests that a substantial fraction of BSI cases might remain undetected; reporting observed BSI rates as a quality indicator without sufficiently high blood culture rates might be misleading.


Binding of ciprofloxacin by humic substances: a molecular dynamics study.
A comprehensive assessment of the potential impacts of antimicrobials released into the environment requires an understanding of their sequestration by natural particles. Of particular interest are the strong interactions of antimicrobials with natural organic matter (NOM), which are believed to reduce their bioavailability, retard their abiotic and biotic degradation, and facilitate their persistence in soils and aquatic sediments. Molecular dynamics (MD) relaxation studies of a widely used fluoroquinolone antibiotic, ciprofloxacin (Cipro), interacting with a model humic substance (HS) in a hydrated environment, were performed to elucidate the mechanisms of these interactions. Specifically, a zwitterionic Cipro molecule, the predominant species at circumneutral pH, was reacted either with protonated HS or deprotonated HS bearing Ca, Mg, or Fe(II) cations. The HS underwent conformational changes through rearrangements of its hydrophobic and hydrophilic regions and disruption of its intramolecular H-bonds to facilitate favorable intermolecular H-bonding interactions with Cipro. Complexation of the metal cations with HS carboxylates appeared to impede binding of the positively charged amino group of Cipro with these negatively charged HS complexation sites. On the other hand, an outer-sphere complex between Cipro and the HS-bound cation led to ternary Cipro-metal-HS complexes in the case of Mg-HS and Fe(II)-HS, but no such bridging interaction occurred with Ca-HS. The results suggested that the ionic potential (valence/ionic radius) of the divalent cation may be a determining factor in the formation of the ternary complex, with high ionic potential favoring the bridging interaction. Environ. Toxicol. Chem. 2010;29:90-98. (c) 2009 SETAC.


SV2 is the protein receptor for botulinum neurotoxin A.
How the widely used botulinum neurotoxin A (BoNT/A) recognizes and enters neurons is poorly understood. We found that BoNT/A enters neurons by binding to the synaptic vesicle protein SV2 (isoforms A, B, and C). Fragments of SV2 that harbor the toxin interaction domain inhibited BoNT/A from binding to neurons. BoNT/A binding to SV2A and SV2B knockout hippocampal neurons was abolished and was restored by expressing SV2A, SV2B, or SV2C. Reduction of SV2 expression in PC12 and Neuro-2a cells also inhibited entry of BoNT/A, which could be restored by expressing SV2 isoforms. Finally, mice that lacked an SV2 isoform (SV2B) displayed reduced sensitivity to BoNT/A. Thus, SV2 acts as the protein receptor for BoNT/A.


Autoregulation of cerebral blood flow during controlled hypotension in baboons.
The effect of graded, progressive hypotension on the autoregulation of cerebral blood flow was studied in anaesthetised baboons. Progressive hypotension was produced over a period of four to five hours, either by graded haemorrhage or by the administration of increasing concentrations of hypotensive drugs. During haemorrhagic hypotension autoregulation was maintained until the mean arterial pressure had decreased to 65% of its baseline value, below which cerebral blood flow was pressure passive. In those animals subjected to drug-induced hypotension, autoregulation persisted to lower levels of mean arterial pressure (35-40% of baseline). It is postulated that under conditions of haemorrhagic hypotension, constriction of the extraparenchymal cerebral vessels in response to sympathetic stimulation decreases the possible range of autoregulation in the anaesthetised baboon.


Secretion of extracellular proteins by enterohemorrhagic Escherichia coli via a putative type III secretion system.
Enterohemorrhagic and enteropathogenic Escherichia coli (EHEC and EPEC) infections result in attaching and effacing lesions on intestinal epithelial cells. Secretion of extracellular proteins via a type III secretion apparatus is necessary for the formation of attaching and effacing lesions by EPEC. We now show that EHEC also secretes polypeptides via a putative type III secretion system. The secreted EHEC proteins are recognized by rabbit antiserum raised against the proteins secreted from EPEC and by human serum from a patient infected with an EHEC O157:H7 strain.


Development of modified intraoperative examination monitor for awake surgery (IEMAS) system for awake craniotomy during brain tumor resection.
Gliomas represent the most frequent type of primary intracranial tumors, which originate from the brain tissue itself, have infiltrative growth, unclear borders, and usually affect functionally-important cerebral structures. From March 2000 till March 2010, 839 neurosurgical procedures directed on resection of such neoplasms were performed in the intelligent operating theater of Tokyo Women's Medical University with the use of intraoperative MRI, real-time updated neuronavigation system, and Hi-vision operative microscope. To facilitate maximal possible tumor resection with minimal risk of neurological morbidity a special device, called Intraoperative Examination Monitor for Awake Surgery (IEMAS) was developed by us. It provides an opportunity to visualize a wide spectrum of inraoperative information related to condition of the patient, nuances of the surgical procedure, and details of the cortical mapping. The wide set of both anatomical and functional parameters, such as view of the patient's mimic and face movements during answering on the specific questions, type of examination test, position of the surgical instruments, parameters of the bispectral index monitor, and general view of the surgical field through the operating microscope and/or endoscope, is presented compactly in one screen with several displays, which allows fast integrated real-time analysis of the multiple data, nearly without interruption of the surgical manipulations. All members of the surgical team can share this information using several in-room liquid crystal displays. However, the initially designed IEMAS system was occasionally affected by interruption or detachment of the connecting cables, which could interfere with effective advancement of the surgical procedure. To avoid this problem a modified device was created. Its specific feature is wireless information transmitting function attained by incorporation of transmitters with a frequency range of 2.4 GHz. The clinical testing of t- - his system was initiated on February 1, 2010, but quickly revealed crossed line effect between transmitters and receivers. To overcome this obstacle and to isolate transmitters, one channel was changed from wireless connection into wired, which resulted in significant improvement of the clearness of both transmitted images and sounds, and provides an opportunity for effective clinical use of the device. In perspective we wish to make IEMAS system fully wireless, using several types of frequency range transmitters.


Smokers with IDDM experience excess morbidity. The Colorado IDDM Registry.
OBJECTIVE--To determine whether there is an association between smoking and the self-reported morbidity of people with IDDM and to evaluate the nature of a possible interaction between smoking and IDDM in increasing the risk of morbidity among smokers with IDDM. RESEARCH DESIGN AND METHODS--Subjects were non-Hispanic whites aged 18-28 yr who participated in the Colorado IDDM Registry Follow-up Survey (case subjects, n = 24) or the 1985 NHIS (control subjects, n = 5876). Assessments of self-reported morbidity included any hospitalization in the past year; bed days, sick days, and limited-activity days in the past 2 wk; and ratings of poor health. The criteria outlined by Saracci were used to determine whether smoking was associated with greater morbidity among IDDM case compared to control subjects (smoking by IDDM interaction). RESULTS--Age- and sex-adjusted ORs, estimated from logistic regression, showed that people with IDDM reported excess morbidity compared with control subjects, regardless of smoking status. Smokers with IDDM reported morbidity 3-10 times as often as nonsmoking control subjects and were 2-3 times more likely to report morbidity than nonsmokers with IDDM. The smoking by IDDM interaction was more than multiplicative for all morbidity measures. Fifty to 75% of excess morbidity in young smokers with IDDM over simple additive effects was related to the interaction between smoking and IDDM. CONCLUSIONS--There was excess reported morbidity among people with IDDM who smoked, greater than that expected from the combined effects of smoking and IDDM. Smoking cessation in young people with IDDM may alleviate some of this excess, but more study is needed to determine whether smoking serves as an indicator of poor IDDM care practices or has a physiological impact that compounds the morbidity experienced by people with IDDM.


Detecting Genuine and Deliberate Displays of Surprise in Static and Dynamic Faces.
People are good at recognizing emotions from facial expressions, but less accurate at determining the authenticity of such expressions. We investigated whether this depends upon the technique that senders use to produce deliberate expressions, and on decoders seeing these in a dynamic or static format. Senders were filmed as they experienced genuine surprise in response to a jack-in-the-box (Genuine). Other senders faked surprise with no preparation (Improvised) or after having first experienced genuine surprise themselves (Rehearsed). Decoders rated the genuineness and intensity of these expressions, and the confidence of their judgment. It was found that both expression type and presentation format impacted decoder perception and accurate discrimination. Genuine surprise achieved the highest ratings of genuineness, intensity, and judgmental confidence (dynamic only), and was fairly accurately discriminated from deliberate surprise expressions. In line with our predictions, Rehearsed expressions were perceived as more genuine (in dynamic presentation), whereas Improvised were seen as more intense (in static presentation). However, both were poorly discriminated as not being genuine. In general, dynamic stimuli improved authenticity discrimination accuracy and perceptual differences between expressions. While decoders could perceive subtle differences between different expressions (especially from dynamic displays), they were not adept at detecting if these were genuine or deliberate. We argue that senders are capable of producing genuine-looking expressions of surprise, enough to fool others as to their veracity.


Oxygen stress reduces zoospore survival of Phytophthora species in a simulated aquatic system.
The genus Phytophthora includes a group of agriculturally important pathogens and they are commonly regarded as water molds. They produce motile zoospores that can move via water currents and on their own locomotion in aquatic environments. However, zoosporic response to dissolved oxygen, an important water quality parameter, is not known. Like other water quality parameters, dissolved oxygen concentration in irrigation reservoirs fluctuates dramatically over time. The aim of this study was to determine whether and how zoospore survival may be affected by elevated and low concentrations of dissolved oxygen in water to better understand the aquatic biology of these pathogens in irrigation reservoirs. Zoospores of P. megasperma, P. nicotianae, P. pini and P. tropicalis were assessed for survival in 10% Hoagland's solution at a range of dissolved concentrations from 0.9 to 20.1 mg L(-1) for up to seven exposure times from 0 to 72 h. Zoospore survival was measured by resultant colony counts per ml. Zoospores of these species survived the best in control Hoagland's solution at dissolved oxygen concentrations of 5.3 to 5.6 mg L(-1). Zoospore survival rates decreased with increasing and decreasing concentration of dissolved oxygen, depending upon Phytophthora species and exposure time. Overall, P. megasperma and P. pini are less sensitive than P. nicotianae and P. tropicalis to hyperoxia and hypoxia conditions. Zoospores in the control solution declined over time and this natural decline process was enhanced under hyperoxia and hypoxia conditions. These findings suggest that dramatic fluctuations of dissolved oxygen in irrigation reservoirs contribute to the population decline of Phytophthora species along the water path in the same reservoirs. These findings advanced our understanding of the aquatic ecology of these pathogens in irrigation reservoirs. They also provided a basis for pathogen risk mitigation by prolonging the turnover time of runoff water in recycling irrigation systems via better system designs.


Characteristics of Sexually Abused Children and Their Nonoffending Mothers Followed by Child Welfare Services: The Role of a Maternal History of Child Sexual Abuse.
Considering the importance of mother's support in the adaptation of a sexually abused child, it is relevant to determine if the mothers and children involved in an intergenerational cycle of child sexual victimization differ from dyads in which only the child has been abused. The purpose of this study was to compare mother-child dyads with sexually abused children according to whether the mother had herself been victim of child sexual abuse. The sample included 87 dyads with sexually abused children aged 3-18 years old and their mothers (44 reporting maternal and child abuse), followed by social welfare services of the province of Quebec (Canada). The two groups of mothers were compared on their past family abuse experiences and past family relations, their mental health history, their current psychological distress, their parenting behaviors, and their current levels of family functioning. Children were compared on their adaptation. Multivariate analyses indicated that mothers reporting child sexual abuse were more likely to report more other maltreatments in their childhood and greater prevalence of lifetime history of alcohol abuse disorders, dysthymia, and panic disorder compared with mothers who had not experienced CSA. Compared to children whose mothers had not experienced CSA, those whose mothers had experienced CSA showed higher rates of problems behaviors and were more likely to report having been sexually abused by a trusted person. These results highlight the specific clinical needs for the assessment and treatment for sexually abused children whose mothers experienced child sexual abuse.


A mechanistic mathematical model for the catalytic action of glutathione peroxidase.
Glutathione peroxidase (GPx) is a well-known seleno-enzyme that protects cells from oxidative stress (e.g., lipid peroxidation and oxidation of other cellular proteins and macromolecules), by catalyzing the reduction of harmful peroxides (e.g., hydrogen peroxide: H₂O₂) with reduced glutathione (GSH). However, the catalytic mechanism of GPx kinetics is not well characterized in terms of a mathematical model. We developed here a mechanistic mathematical model of GPx kinetics by considering a unified catalytic scheme and estimated the unknown model parameters based on different experimental data from the literature on the kinetics of the enzyme. The model predictions are consistent with the consensus that GPx operates via a ping-pong mechanism. The unified catalytic scheme proposed here for GPx kinetics clarifies various anomalies, such as what are the individual steps in the catalytic scheme by estimating their associated rate constant values and a plausible rationale for the contradicting experimental results. The developed model presents a unique opportunity to understand the effects of pH and product GSSG on the GPx activity under both physiological and pathophysiological conditions. Although model parameters related to the product GSSG were not identifiable due to lack of product-inhibition data, the preliminary model simulations with the assumed range of parameters show that the inhibition by the product GSSG is negligible, consistent with what is known in the literature. In addition, the model is able to simulate the bi-modal behavior of the GPx activity with respect to pH with the pH-range for maximal GPx activity decreasing significantly as the GSH levels decrease and H₂O₂ levels increase (characteristics of oxidative stress). The model provides a key component for an integrated model of H₂O₂ balance under normal and oxidative stress conditions.


A mathematical model for targeting chemicals to tissues by exploiting complex degradation.
In many biological and therapeutic contexts, it is highly desirable to target a chemical specifically to a particular tissue where it exerts its biological effect. In this paper, we present a simple, generic, mathematical model that elucidates a general method for targeting a chemical to particular tissues. The model consists of coupled reaction-diffusion equations to describe the evolution within the tissue of the concentrations of three chemical species: a (concentration of free chemical), b (binding protein) and their complex, c (chemical bound to binding protein). We assume that all species are free to diffuse, and that a and b undergo a reversible reaction to form c. In addition, the complex, c, can be broken down by a process (e.g. an enzyme in the tissue) that results in the release of the chemical, a, which is then free to exert its biological action. For simplicity, we consider a one-dimensional geometry. In the special case where the rate of complex formation is small (compared to the diffusion timescale of the species within the tissue) the system can be solved analytically. This analytic solution allows us to show how the concentration of free chemical, a, in the tissue can be increased over the concentration of free chemical at the tissue boundary. We show that, under certain conditions, the maximum concentration of a can occur at the centre of the tissue, and give an upper bound on this maximum level. Numerical simulations are then used to determine how the behaviour of the system changes when the assumption of negligible complex formation rate is relaxed. We have shown, using our mathematical model, how complex degradation can potentially be exploited to target a chemical to a particular tissue, and how the level of the active chemical depends on factors such as the diffusion coefficients and degradation/production rates of each species. The biological significance of these results in terms of potential applications in cartilage tissue engineering and chemotherapy is discussed. In particular, we believe these results may be of use in determining the most promising prodrug candidates.


WE-G-217A-01: Fast Recovery Driven Equilibrium with Automated Switching to Avoid Artifacts.
To reduce image artifacts from the unnecessary use of driven- equilibrium. Driven-equilibrium methods known as fast recovery, RESTORE, or DRIVE enable reduction of repetition time (TR) and hence acquisition time in RARE (FSE or TSE) imaging by applying additional RF pulses following each echotrain to accelerate T1 recovery. Fast recovery (FR) techniques are most useful when TR is less than 4*T1. For neuroimaging, the benefit of FR diminishes as TR increases beyond ∼3s. In regions of rapid susceptibility change such as near metallic implants, FR techniques can introduce artifact because the magnetization is not properly rephased when the FR pulses are applied. In our clinical practice we observe that technologists frequently adjust TR over a range of 2-5s on a patient- specific basis to minimize scan time. We do not observe, however, that technologists reliably disable FR when TR is long enough, introducing possible artifacts. An FR-FSE pulse sequence was modified to automatically disable the FR feature whenever TR exceeded a user-defined threshold (TR0). Under IRB approval, a healthy volunteer was scanned with the sequence using TRs ranging from 1-5s, and TR0=900ms or 6s. Contrast- to-noise measurements were made between CSF, gray matter and white matter for each dataset. The volunteer study shows decreasing benefit of FR as TRs increase beyond 3s. A patient with bilateral auditory brainstem implants scanned with and without FR demonstrates FR-induced artifacts. We demonstrate an FR-FSE pulse sequence that automatically disables fast-recovery whenever TR exceeds an operator- selected threshold. The pulse sequence avoids unnecessary artifacts in regions of rapid susceptibility changes, while retaining the advantages of FR when TR<TR0. Clinical images from a patient with bilateral auditory brainstem implants demonstrate potential artifacts introduced by FR, and the benefits of automatically avoiding unnecessary use of FR.


Synaptic localization of P2X7 receptors in the rat retina.
The distribution of P2X(7) receptor (P2X(7)R) subunits was studied in the rat retina using a subunit-specific antiserum. Punctate immunofluorescence was observed in the inner and outer plexiform layers. Double labeling of P2X(7) and the horizontal cell marker, calbindin, revealed extensive colocalization in the outer plexiform layer (OPL). Significant colocalization of P2X(7)R and kinesin, a marker of photoreceptor ribbons, was also observed, indicating that this receptor may be expressed at photoreceptor terminals. Furthermore, another band of P2X(7)R puncta was identified below the level of the photoreceptor terminals, adjacent to the inner nuclear layer (INL). This band of P2X(7)R puncta colocalized with the active-zone protein, bassoon, suggesting that "synapse-like" structures exist outside photoreceptor terminals. Preembedding immunoelectron microscopy demonstrated P2X(7)R labeling of photoreceptor terminals adjacent to ribbons. In addition, some horizontal cell dendrites and putative "desmosome-like" junctions below cone pedicles were labeled. In the inner plexiform layer (IPL), P2X(7)R puncta were observed surrounding terminals immunoreactive for protein kinase C-alpha, a marker of rod bipolar cells. Double labeling with bassoon in the IPL revealed extensive colocalization, indicating that P2X(7)R is likely to be found at conventional cell synapses. This finding was confirmed at the ultrastructural level: only processes presynaptic to rod bipolar cells were found to be labeled for the P2X(7)R, as well as other conventional synapses. These findings suggest that purines play a significant role in neurotransmission within the retina, and may modulate both photoreceptor and rod bipolar cell responses.


Differences in the ventilatory responses of horses and ponies to exercise of varying intensities.
Horses exercising at > or = approximately 90% VO2max develop arterial hypoxaemia with concurrent hypercapnoea, whereas ponies exercising at comparative levels become hypocapnoeic and maintain arterial oxygen tensions close to resting values. We sought to investigate the possibility that these differences relate to the ventilatory responses of these animals to exercise. Six Thoroughbred horses weighing mean +/- s.e. 501 +/- 27 kg and 5 ponies weighing mean +/- s.e. 164 +/- 18 kg exercised for 2 min on a 10% slope at speeds calculated to require 60% VO2max and for at least 1 min at speeds calculated to require 115% VO2max. Oxygen consumption (VO2), arterial oxygen (PaO2) and carbon dioxide (PaCO2) tensions, acid-base balance, tidal volume (VT), minute ventilation (VE), peak inspiratory (VImax) and expiratory (VEmax) flow, and maximal changes in transpulmonary pressures (delta PtPmax) were measured immediately before exercise and in the last 15 s of exercise at each intensity. The results confirmed that, unlike horses, ponies do not become hypoxaemic or hypercapnoeic during exercise. Despite having a higher delta PtPmax, higher VImax and VEmax and VE/kg0.75 at the same relative intensities, horses were less capable of mounting an appropriate ventilatory response to exercise. This was reflected by lower mass specific and metabolic weight-based ventilations at similar absolute workloads, and their higher PaCO2 and arterial HCO3-[, and lower ventilatory equivalent for oxygen (VE/VO2). This suggests that horses become hypoxaemic and hypercapnoeic at work loads > or = approximately 90% VO2max because their metabolic demand surpasses the capacity of their ventilatory system to meet this demand. Because ponies are less capable athletes, they can match their ventilatory response to their metabolic requirements.


The rutin catabolic pathway with special emphasis on quercetinase.
The aim of this review is to give a general account on the oxidative microbial degradation of flavonols. Since now 50 years, various research groups have deciphered the way microorganisms aerobically deal with this important class of flavonoids. Flavonols such as rutin and quercetin are abundantly found in vegetal tissues and exudates, and it was thus patent that various microorganisms will bear the enzymatic machinery necessary to cope with these vegetal secondary metabolites. After initial studies focussed on the general metabolic capacity of various microorganisms towards flavonols, the so called rutin catabolic pathway was rapidly established in moulds. Enzymes of the path as well as substrates and products were known at the beginning of the seventies. Then during 30 years, only sporadic studies were focused on this pathway, before a new burst of interest at the beginning of the new century arose with structural, genomic and theoretical studies mainly conducted towards quercetinase. This is the goal of this work to relate this 50 years journey at the crossroads of microbiology, biochemistry, genetic and chemistry. Some mention of the potential usefulness of the enzymes of the path as well as micro-organisms bearing the whole rutin catabolic pathway is also discussed.


DNA repair and cell cycle biomarkers of radiation exposure and inflammation stress in human blood.
DNA damage and repair are hallmarks of cellular responses to ionizing radiation. We hypothesized that monitoring the expression of DNA repair-associated genes would enhance the detection of individuals exposed to radiation versus other forms of physiological stress. We employed the human blood ex vivo radiation model to investigate the expression responses of DNA repair genes in repeated blood samples from healthy, non-smoking men and women exposed to 2 Gy of X-rays in the context of inflammation stress mimicked by the bacterial endotoxin lipopolysaccharide (LPS). Radiation exposure significantly modulated the transcript expression of 12 genes of 40 tested (2.2E-06<p<0.03), of which 8 showed no overlap between unirradiated and irradiated samples (CDKN1A, FDXR, BBC3, PCNA, GADD45a, XPC, POLH and DDB2). This panel demonstrated excellent dose response discrimination (0.5 to 8 Gy) in an independent human blood ex vivo dataset, and 100% accuracy for discriminating patients who received total body radiation. Three genes of this panel (CDKN1A, FDXR and BBC3) were also highly sensitive to LPS treatment in the absence of radiation exposure, and LPS co-treatment significantly affected their radiation responses. At the protein level, BAX and pCHK2-thr68 were elevated after radiation exposure, but the pCHK2-thr68 response was significantly decreased in the presence of LPS. Our combined panel yields an estimated 4-group accuracy of ∼90% to discriminate between radiation alone, inflammation alone, or combined exposures. Our findings suggest that DNA repair gene expression may be helpful to identify biodosimeters of exposure to radiation, especially within high-complexity exposure scenarios.


Quantitative carbon detector for enhanced detection of molecules in foods, pharmaceuticals, cosmetics, flavors, and fuels.
Analysis of trace compounds, such as pesticides and other contaminants, within consumer products, fuels, and the environment requires quantification of increasingly complex mixtures of difficult-to-quantify compounds. Many compounds of interest are non-volatile and exhibit poor response in current gas chromatography and flame ionization systems. Here we show the reaction of trimethylsilylated chemical analytes to methane using a quantitative carbon detector (QCD; the Polyarc™ reactor) within a gas chromatograph (GC), thereby enabling enhanced detection (up to 10×) of highly functionalized compounds including carbohydrates, acids, drugs, flavorants, and pesticides. Analysis of a complex mixture of compounds shows that the GC-QCD method exhibits faster and more accurate analysis of complex mixtures commonly encountered in everyday products and the environment.


Inhibition of benzo(a)pyrene-induced forestomach carcinogenesis in mice by thymoquinone.
The modulating effect of thymoquinone (TQ) on benzo(a)pyrene (BP)-induced forestomach tumours was investigated in female Swiss albino mice, receiving oral administration of BP at a dose of 1 mg twice weekly for 4 weeks. Administration of 0.01% of TQ in drinking water 1 week before, during and after BP treatment until the end of the experiment resulted in significant suppression of BP-induced tumourigenesis when compared with the group receiving BP alone. TQ inhibited both BP-induced forestomach tumour incidence and multiplicity by 70% and 67%, respectively. Lipid peroxide accumulation and decreased glutathione (GSH) content and glutathione-S-transferase (GST) and DT diaphorase activities were observed in the liver of BP-treated tumour-bearing mice. TQ alone showed a significant induction in the enzyme activities of hepatic GST and DT diaphorase. Mice treated with TQ along with BP showed almost normal hepatic lipid peroxides and GSH levels, and normal enzyme activities compared to the control group. The present data may indicate the potential of TQ, the main constituent of the volatile oil of Nigella sativa seed, as a powerful chemopreventive agent against BP-induced forestomach tumours in mice. The possible modes of action of TQ may be through its antioxidant and anti-inflammatory activities, coupled with enhancement of detoxification processes.


Gelatin-tricalcium phosphate membranes immobilized with NGF, BDNF, or IGF-1 for peripheral nerve repair: an in vitro and in vivo study.
In the present study, NGF, BNDF from the neurotrophin family and IGF-1 were covalently immobilized on gelatin-tricalcium phosphate (GTG) membrane using carbodiimide. We investigated the effects of these growth factors released from the GTG composites on cultured PC12 cells and sciatic nerve regeneration across a 10-mm-long gap in rats. In PC12 cell culture, the total protein content and MTT assay indicated more cell attachment on the composites modified with growth factors. The IGF-1 group showed a higher survival promotion effect on PC12 cells than did BDNF and NGF groups. On the other hand, NGF released from the composite showed the highest level of neuritogenesis for PC12 cells in neurite outgrowth assay. In the animal study, the GTG conduits modified with various growth factors were well tolerated by the host tissue. In the regenerated nerves, the number of the axons per unit area of the BDNF group was significantly higher than that of NGF and GTG groups but similar to that of IGF-1 group. However, the average axon size was the largest in NGF group. This result was in concordance with the neurite outgrowth assay in which NGF showed the highest neuritogenic potential. In the assessment of motor and sensory recovery after nerve repair, conduits modified with various neurotrophic factors showed a more favorable outcome in compound muscle action potential. The BDNF group had a better gastrocnemic muscle weight ratio than blank GTG repair. Nevertheless, the different effects of GTG conduits modified with various neurotrophic factors on functional recovery cannot be simply illustrated in the sciatic function index.


Advancing general practice nursing in Australia: roles and responsibilities of primary healthcare organisations.
Objectives Given increased numbers and enhanced responsibilities of Australian general practice nurses, we aimed to delineate appropriate roles for primary health care organisations (PHCOs) to support this workforce. Methods A two-round online Delphi consensus process was undertaken between January and June 2012, informed by literature review and key informant interviews. Participants were purposively selected and included decision makers from government and professional organisations, educators, researchers and clinicians from five Australian states and territories Results Of 56 invited respondents, 35 (62%) and 31 (55%) responded to the first and second invitation respectively. Participants reached consensus on five key roles for PHCOs in optimising nursing in general practice: (1) matching workforce size and skills to population needs; (2) facilitating leadership opportunities; (3) providing education and educational access; (4) facilitating integration of general practice with other primary care services to support interdisciplinary care; and (5) promoting advanced nursing roles. National concerns, such as limited opportunities for postgraduate education and career progression, were deemed best addressed by national nursing organisations, universities and peak bodies. Conclusions Advancement of nursing in general practice requires system-level support from a range of organisations. PHCOs play a significant role in education and leadership development for nurses and linking national nursing organisations with general practices. What is known about the topic? The role of nurses in Australian general practice has grown in the last decade, yet they face limited career pathways and opportunities for career advancement. Some nations have forged interprofessional primary care teams that use nurses' skills to the full extent of their scope of practice. PHCOs have played important roles in the development of general practice nursing in Australia and internationally. What does this paper add? This study delineates organisational support roles for PHCOs in strengthening nurses' roles and career development in Australian general practice. What are the implications for practitioners? Effective implementation of appropriate responsibilities by PHCOs can assist development of the primary care nursing workforce.


Successful treatment with clarithromycin and/or tacrolimus for two patients with polymyalgia rheumatica.
Polymyalgia rheumatica (PMR) is an inflammatory rheumatic disease in the elderly. Glucocorticoids (GCs) remain the mainstay of treatment. GC therapy usually dramatically improves the clinical picture, but approximately one-third of patients experience disease recurrence when the dose is reduced. Moreover, long-term use of GCs causes adverse reactions. Macrolide antibiotics have anti-inflammatory action. Several recent studies have reported the successful treatment of rheumatoid arthritis (RA) and PMR treated using clarithromycin (CAM), a macrolide, because of its anti-inflammatory action. Tacrolimus (TAC) has been indicated as a treatment for RA in patients who failed to respond to methotrexate. Recently, a case of RA was successfully treated using CAM and TAC according to one report. Reported here are two cases of PMR treated using CAM and/or TAC. Case 1: A 73-year-old man suffering from PMR was successfully treated with prednisolone (PSL) and CAM. Because his muscle pain disappeared, CAM was discontinued. However, the pain returned after that discontinuation, so CAM was successfully administered again. Case 2: An 83-year-old man suffering from PMR was successfully treated with PSL and CAM. Because muscle pain disappeared, the CAM dosage was halved. The pain returned after the dosage was reduced, so the CAM dosage was successfully resumed and the PSL dosage was reduced. When the PSL dosage was reduced, muscle pain recurred. Because the PSL and CAM dosages were not successfully increased, TAC was also administered and was found to be effective at treating muscle pain. These two cases suggest that CAM and/or TAC are effective at treating PMR.


Inheritance of Strain Instability (Sectoring) in the Commercial Button Mushroom, Agaricus bisporus.
The button mushroom, Agaricus bisporus, is a commercially important cultivated filamentous fungus. During the last decade, the button mushroom industry has depended mainly on two strains (or derivatives of these two strains). Using one of these highly successful strains (strain U1) we examined the phenomenon of strain instability, specifically, the production of irreversible sectors. Three "stromatal" and three "fluffy" sectors were compared with a healthy type U1 strain and with a wild-collected isolate. Compost colonization and fruit body morphology were examined. The main objective of this study, however, was to examine the meiotic stability of the sectored phenotype. Single basidiospores were isolated and subjected to a grain bioassay in which the ability to produce sectors was measured. Our results were as follows: (i) basidiospore cultures obtained from a wild-collected isolate showed no tendency to produce sectors; (ii) approximately 5% of the basidiospore cultures obtained from healthy type U1 strains produced irreversible sectors in the grain bioassay; (iii) the five primary sectors examined produced basidiospore cultures, half of which produced normal-looking growth in the grain bioassay and half of which produced some degree of sectoring; and (iv) the one sectored isolate that represented the F2 generation gave ratios similar to the 1:1 ratio observed for the F1 cultures.


Abdominal MRI without Enteral Contrast Accurately Detects Intestinal Fibrostenosis in Patients with Inflammatory Bowel Disease.
Patients with inflammatory bowel disease (IBD) presenting for surgical evaluation require thorough small bowel surveillance as it improves accuracy of diagnosis (ulcerative colitis versus Crohn's) and differentiates those who may respond to nonoperative therapy, preserving bowel length. MRI has not been validated conclusively against histopathology in IBD. Most protocols require enteral contrast. This study aimed to 1) evaluate the accuracy of MRI for inflammation, fibrosis, and extraluminal complications and 2) compare MRI without enteral contrast to standard magnetic resonance enterography. Adults with Crohn's disease or ulcerative colitis who underwent abdominal MRI and surgery were retrospectively reviewed. Of 65 patients evaluated, 55 met inclusion criteria. Overall sensitivity and specificity of MRI for disease involvement localized by segment were 93 per cent (95% confidence interval = 89.4-95.0) and 95 per cent (95% confidence interval = 92.3-97.0), respectively (positive predictive value was 86%, negative predictive value was 98%). Sensitivity and specificity between MRI with and without oral and rectal contrast were similar (96% vs 91% and 99% vs 94%, P > 0.10). As were positive predictive value and negative predictive value (85% vs 96%, P = 0.16; 97% vs 99%, P = 0.42). Magnetic resonance is highly sensitive and specific for localized disease involvement and extraluminal abdominal sequelae of IBD. It accurately differentiates patients who have chronic transmural (fibrotic) disease and thus may require an operation from those with acute inflammation, whose symptoms may improve with aggressive medical therapy alone. MRI without contrast had comparable diagnostic yield to standard magnetic resonance enterography.


In vivo delivery of peptides and Toll-like receptor ligands by mannose-functionalized polymeric nanoparticles induces prophylactic and therapeutic anti-tumor immune responses in a melanoma model.
We hypothesized that the co-entrapment of melanoma-associated antigens and the Toll-like receptor (TLR) ligands Poly(I:C) and CpG, known to be Th1-immunopotentiators, in mannose-functionalized aliphatic polyester-based nanoparticles (NPs) could be targeted to mannose receptors on antigen-presenting cells and induce anti-tumor immune responses. High entrapment efficiencies of antigens and immunopotentiators in 150nm NPs were obtained. The co-entrapment of the model antigen ovalbumin and the TLR ligands was crucial to induce high IgG2c/IgG1 ratios and high levels of IFN-γ and IL-2. Mannose-functionalization of NPs potentiated the Th1 immune response. The nanoparticulate vaccines decreased the growth rate of murine B16F10 melanoma tumors in therapeutic and prophylatic settings. The combination of mannose-functionalized NPs containing MHC class I- or class II-restricted melanoma antigens and the TLR ligands induced the highest tumor growth delay. Overall, we demonstrate that the multifunctional properties of NPs in terms of targeting and antigen/adjuvant delivery have high cancer immunotherapeutic potential.


Incidence of cancer among welders and other workers in a Norwegian shipyard.
The incidence of cancer among 4778 male shipyard workers, including 783 mild steel welders, was investigated in a historical cohort study. The workers had been employed for at least three months between 1 January 1946 and 31 March 1977. The incidence of cancer was observed from 1 January 1953 through 1986. The loss during follow-up was only 0.9%. There were 53 observed cases of lung cancer in the whole cohort versus 31.3 expected on the basis of the national rates for men. There was an increased incidence of lung cancer among the welders, with seven observed cases versus 3.2 expected. Twenty-two cases of bladder cancer were observed versus 15.2 expected. Two malignant mesotheliomas had occurred (0.7 expected). Smoking was likely to be a confounder in the present study. Due to concomitant exposure to asbestos, the results are inconclusive concerning the possible relationship between exposure to welding fumes and lung cancer.


Homozygosity mapping and whole exome sequencing reveal a novel homozygous COL18A1 mutation causing Knobloch syndrome.
The aim of this study was to identify the genetic basis of a chorioretinal dystrophy with high myopia of unknown origin in a child of a consanguineous marriage. The proband and ten family members of Iranian ancestry participated in this study. Linkage analysis was carried out with DNA samples of the proband and her parents by using the Human SNP Array 6.0. Whole exome sequencing (WES) was performed with the patients' DNA. Specific sequence alterations within the homozygous regions identified by whole exome sequencing were verified by Sanger sequencing. Upon genetic analysis, a novel homozygous frameshift mutation was found in exon 42 of the COL18A1 gene in the patient. Both parents were heterozygous for this sequence variation. Mutations in COL18A1 are known to cause Knobloch syndrome (KS). Retrospective analysis of clinical records of the patient revealed surgical removal of a meningocele present at birth. The clinical features shown by our patient were typical of KS with the exception of chorioretinal degeneration which is a rare manifestation. This is the first case of KS reported in a family of Iranian ancestry. We identified a novel disease-causing (deletion) mutation in the COL18A1 gene leading to a frameshift and premature stop codon in the last exon. The mutation was not present in SNP databases and was also not found in 192 control individuals. Its localization within the endostatin domain implicates a functional relevance of endostatin in KS. A combined approach of linkage analysis and WES led to a rapid identification of the disease-causing mutation even though the clinical description was not completely clear at the beginning.


Assessing Medicare's hospital pay-for-performance programs and whether they are achieving their goals.
Three separate pay-for-performance programs affect the amount of Medicare payment for inpatient services to about 3,400 US hospitals. These payments are based on hospital performance on specified measures of quality of care. A growing share of Medicare hospital payments (6 percent by 2017) are dependent upon how hospitals perform under the Hospital Readmissions Reduction Program, the Value-Based Purchasing Program, and the Hospital-Acquired Condition Reduction Program. In 2015 four of five hospitals subject to these programs will be penalized under one or more of them, and more than one in three major teaching hospitals will be penalized under all three. Interactions among these programs should be considered going forward, including overlap among measures and differences in scoring performance.


Enhancing Food Processing by Pulsed and High Voltage Electric Fields: Principles and Applications.
Improvements in living standards result in a growing demand for food with high quality attributes including freshness, nutrition and safety. However, current industrial processing methods rely on traditional thermal and chemical methods, such as sterilization and solvent extraction, which could induce negative effects on food quality and safety. The electric fields (EFs) involving pulsed electric fields (PEFs) and high voltage electric fields (HVEFs) have been studied and developed for assisting and enhancing various food processes. In this review, the principles and applications of pulsed and high voltage electric fields are described in details for a range of food processes, including microbial inactivation, component extraction, and winemaking, thawing and drying, freezing and enzymatic inactivation. Moreover, the advantages and limitations of electric field related technologies are discussed to foresee future developments in the food industry. This review demonstrates that electric field technology has a great potential to enhance food processing by supplementing or replacing the conventional methods employed in different food manufacturing processes. Successful industrial applications of electric field treatments have been achieved in some areas such as microbial inactivation and extraction. However, investigations of HVEFs are still in an early stage and translating the technology into industrial applications need further research efforts.


Optical and electronic properties of lithium thiogallate (LiGaS2): experiment and theory.
We report the relation between the optical properties and electronic structure of lithium thiogallate (LiGaS2) by performing XPS and XES measurements and theoretical calculations. According to the XPS measurements, the LiGaS2 crystals grown by the Bridgman-Stockbarger method possess promising optical qualities, low hygroscopicity and high stability upon middle-energy Ar+-ion irradiation. The difference in the LiGaS2 band gaps obtained by theoretical calculations and experimental measurements was, for the first time, reduced down to 0.27 eV by applying the Tran-Blaha modified Becke-Johnson (TB-mBJ) potential where the Coulomb repulsion was considered by introducing Hubbard parameter, U. The TB-mBJ+U method also reproduces the XPS spectrum well. The TB-mBJ+U band-structure calculations of LiGaS2 are found to be in good agreement with the XPS and XES experimental data. The accurate electronic structure of LiGaS2 allows further investigation of the optical properties. The relation between the photoluminescence of LiGaS2 and its electronic structure was revealed. Moreover, the theoretical results show the possibility of emissions at higher energy levels in LiGaS2, that has not been measured in experiments yet. Good phase-matching of LiGaS2 was expected to occur at energy levels of 5, 6, 6.2, 7, 7.2, and 8 eV.


Long-term survival rates of gravity-assisted, adjustable differential pressure valves in infants with hydrocephalus.
OBJECTIVE The use of adjustable differential pressure valves with gravity-assisted units in shunt therapy of children with hydrocephalus was reported to be feasible and promising as a way to avoid chronic overdrainage. In this single-center study, the authors' experiences in infants, who have higher rates of shunt complications, are presented. METHODS All data were collected from a cohort of infants (93 patients [37 girls and 56 boys], less than 1 year of age [mean age 4.1 ± 3.1 months]) who received their first adjustable pressure hydrocephalus shunt as either a primary or secondary implant between May 2007 and April 2012. Rates of valve and shunt failure were recorded for a total of 85 months until the end of the observation period in May 2014. RESULTS During a follow-up of 54.2 ± 15.9 months (range 26-85 months), the Kaplan-Meier rate of shunt survival was 69.2% at 1 year and 34.1% at 85 months; the Kaplan-Meier rate of valve survival was 77.8% at 1 year and 56% at 85 months. Survival rates of the shunt were significantly inferior if the patients had previous shunt surgery. During follow-up, 44 valves were exchanged in cases of infection (n = 19), occlusion (n = 14), dysfunction of the adjustment unit (n = 10), or to change the gravitational unit (n = 1). CONCLUSIONS Although a higher shunt complication rate is observed in infant populations compared with older children, reasonable survival rates demonstrate the feasibility of using this sophisticated valve technology. The gravitational unit of this valve is well tolerated and its adjustability offers the flexible application of opening pressure in an unpredictable cohort of patients. This may adequately address overdrainage-related complications from early in treatment.


Effects of spray drying on physicochemical properties of chitosan acid salts.
The effects of spray-drying process and acidic solvent system on physicochemical properties of chitosan salts were investigated. Chitosan used in spray dryings was obtained by deacetylation of chitin from lobster (Panulirus argus) origin. The chitosan acid salts were prepared in a laboratory-scale spray drier, and organic acetic acid, lactic acid, and citric acid were used as solvents in the process. The physicochemical properties of chitosan salts were investigated by means of solid-state CP-MAS (13)C nuclear magnetic resonance (NMR), X-ray powder diffraction (XRPD), differential scanning calorimetry, and Fourier transform infrared spectrometry (FTIR) and near-infrared spectroscopy. The morphology of spray-dried chitosan acid salts showed tendency toward higher sphericity when higher temperatures in a spray-drying process were applied. Analysis by XRPD indicated that all chitosan acid salts studied were amorphous solids. Solid-state (13)C NMR spectra revealed the evidence of the partial conversion of chitosan acetate to chitin and also conversion to acetyl amide form which appears to be dependent on the spray-drying process. The FTIR spectra suggested that the organic acids applied in spray drying may interact with chitosan at the position of amino groups to form chitosan salts. With all three chitosan acid salts, the FTIR bands at 1,597 and 1,615 cm(-1) were diminished suggesting that -NH groups are protonated. The FTIR spectra of all chitosan acid salts exhibited ammonium and carboxylate bands at 1,630 and 1,556 cm(-1), respectively. In conclusion, spray drying is a potential method of preparing acid salts from chitosan obtained by deacetylation of chitin from lobster (P. argus) origin.


Working memory and intelligence--their correlation and their relation: comment on Ackerman, Beier, and Boyle (2005).
On the basis of a meta-analysis of pairwise correlations between working memory tasks and cognitive ability measures, P. L. Ackerman, M. E. Beier, and M. O. Boyle (2005; see record 2004-22408-002) claimed that working memory capacity (WMC) shares less than 25% of its variance with general intelligence (g) and with reasoning ability. In this comment, the authors argue that this is an underestimation because of several methodological shortcomings and biases. A reanalysis of the data reported in Ackerman et al. using the correct statistical procedures demonstrates that g and WMC are very highly correlated. On a conceptual level, the authors point out that WMC should be regarded as an explanatory construct for intellectual abilities. Theories of working memory do not claim that WMC is isomorphic with intelligence factors but that it is a very strong predictor of reasoning ability and also predicts general fluid intelligence and g.


Communication of professional literature amongst European Acupuncturists affiliated to the ETCMA (European Traditional Chinese Medicine Association): explorative survey amongst Traditional Chinese Medicine practitioners in Europe.
The primary aim of the survey was to explore the information needs and information seeking behavior amongst the ETCMA members concerning professional literature (scientific as well as practical background knowledge). A web-based survey comprising of 18 questions with a total of 25 items was carried out in 15 affiliated associations in 14 countries in June 2012. The survey consisted out of 4 parts: (1) Demographics, (2) Level of interest in and availability of professional literature, (3) Insight, needs and opinions on EBM (Evidence Based Medicine), and (4) Awareness of the science workshop at the TCM Rothenburg Congress. 2590 (25%) from 10,428 members completed the questionnaire, of which 58.8% was female. More than 50% of the respondents from eleven out of fourteen countries indicate an interest in more education on reading scientific literature. Case studies (range 3.19/4-3.86/4) are preferred compared to scientific (range 2.78/4-3.59/4) or philosophical knowledge (range 3.0/4-3.56/4). Exchange with colleagues (range 2.95/4-3.64/4) is preferred compared to deepening knowledge (range 2.57/4-3.05/4) in the theoretical spectrum. 61% has no knowledge of the EBM model and base clinical decisions on personal experience (range 3.47-3.82) and practical skills (range 3.47-3.74) compared to clinical practice guidelines (range 2.6-3.27). Due to heterogeneity in structure and size of the affiliated associations no strict conclusions can be made. We can conclude though that TCM practitioners rely mostly on practical knowledge and have less tendency toward more scientifically oriented models like the EBM model. We find this reflected in information needs as well as information seeking behavior patterns.


Microbial diversity within early-stage cultured Panulirus ornatus phyllosomas.
A thorough understanding of the microorganisms and pathogens associated with the larval stage of the tropical ornate rock lobster, Panulirus ornatus, is required to overcome disease outbreaks that currently block aquaculture attempts. This study used microscopy in addition to culture and molecularly based microbiological techniques to characterize the bacterial community associated with cultured, developmental stage PI to PII P. ornatus phyllosomas. Scanning electron microscopy demonstrated colonization of phyllosomas by filamentous, rod-shaped, and coccus-shaped bacteria. A clone library constructed from dead phyllosomas sampled from the larval rearing tank on day 10 was dominated by Thiothrix-affiliated sequences (56% of clones). A comparable library from live phyllosomas also contained Thiothrix-affiliated sequences, though these only represented 19% of clones within the library. Fluorescent in situ hybridization (FISH) confirmed identification of the filamentous bacteria as Thiothrix sp., being present on dead phyllosomas. FISH also identified Leucothrix sp. and Vibrio sp., as well as a range of other rod- and coccus-shaped bacteria, colonizing both live and dead phyllosomas. The development of the microbial community associated with phyllosomas was monitored through a standard larval rearing run using denaturing gradient gel electrophoresis (DGGE). Vibrio sp.-affiliated bands dominated the profiles of live animals through the rearing period and dead phyllosomas sampled on selected days. The population of Vibrio sp. associated with phyllosomas was monitored with culture-based analysis on selective media and demonstrated to increase significantly on day 7, coinciding with the beginning of the larval molt. An isolated Vibrio harveyi strain demonstrated an identical 16S rRNA sequence with retrieved DGGE and clone library sequences. Colonization of phyllosomas with filamentous bacterial species potentially hinders the ability of the animals to molt and, combined with the added stress of the molt process, likely results in reduced immune function, allowing opportunistic pathogenic Vibrio sp. to cause larval mortalities.


Novel minicircle vector for gene therapy in murine myocardial infarction.
Conventional plasmids for gene therapy produce low-level and short-term gene expression. In this study, we develop a novel nonviral vector that robustly and persistently expresses the hypoxia-inducible factor-1 alpha (HIF-1alpha) therapeutic gene in the heart, leading to functional benefits after myocardial infarction. We first created minicircles (MC) carrying double-fusion reporter gene consisting of firefly luciferase and enhanced green fluorescent protein (Fluc-eGFP) for noninvasive measurement of transfection efficiency. Mouse C2C12 myoblasts and normal FVB/N mice were used for in vitro and in vivo confirmation, respectively. Bioluminescence imaging showed stable MC gene expression in the heart for >12 weeks and the activity level was 5.6+/-1.2-fold stronger than regular plasmid at day 4 (P<0.01). Next, we created MC carrying HIF-1alpha (MC-HIF-1alpha) therapeutic gene for treatment of myocardial infarction. Adult FVB/N mice underwent left anterior descending ligation and were injected intramyocardially with: (1) MC-HIF-1alpha; (2) regular plasmid carrying HIF-1alpha (PL-HIF-1alpha) as positive control; and (3) PBS as negative control (n=10/group). Echocardiographic study showed a significantly greater improvement of left ventricular ejection fraction in the MC group (51.3%+/-3.6%) compared to regular plasmid group (42.3%+/-4.1%) and saline group (30.5%+/-2.8%) at week 4 (P<0.05 for both). Histology demonstrated increased neoangiogenesis in both treatment groups. Finally, Western blot showed MC express >50% higher HIF-1alpha level than regular plasmid. Taken together, this is the first study to our knowledge to demonstrate that MC can significantly improve transfection efficiency, duration of transgene expression, and cardiac contractility. Given the serious drawbacks associated with most viral vectors, we believe this novel nonviral vector can be of great value for cardiac gene therapy protocols.


Identifying residual foci of Plasmodium falciparum infections for malaria elimination: the urban context of Khartoum, Sudan.
Identifying the location and size of residual foci of infections is critical where malaria elimination is the primary goal. Here the spatial heterogeneity of Plasmodium falciparum infections within the urban extent of Khartoum state in Sudan is investigated using data from cross-sectional surveys undertaken from 1999 to 2008 to inform the Khartoum Malaria Free Initiative (KMFI). From 1999-2008 the KMFI undertook cross-sectional surveys of 256 clusters across 203 random samples of residential blocks in the urban Khartoum state in September of each year. Within sampled blocks, at least five persons, including at least one child under the age of five years, were selected from each household. Blood smears were collected from the sampled individuals to examine the presence of P. falciparum parasites. Residential blocks were mapped. Data were analysed for spatial clustering using the Bernoulli model and the significance of clusters were tested using the Kulldorff scan statistic. A total of 128,510 malaria slide examinations were undertaken during the study period. In 1999, overall prevalence was 2.5%, rising to 3.2% in 2000 and consistently staying below 1% in subsequent years. From 2006, over 90% of all surveyed clusters reported no infections. Spatial clustering of infections was present in each year but not statistically significant in the years 2001, 2002, 2004 and 2008. Spatial clusters of high infection were often located at the junction of the Blue and White Niles. Persisting foci of malaria infection in Khartoum are likely to distort wide area assessments and disproportionately affect future transmission within the city limits. Improved investments in surveillance that combines both passive and active case detection linked to a geographic information system and a more detailed analysis of the location and stability of foci should be undertaken to facilitate and track malaria elimination in the state of Khartoum.


Measurement of complex Chi((3)) using degenerate four-wave mixing with an imaged 2-D phase grating.
We present a simple optical arrangement for phase sensitive detection of degenerate four-wave mixing (DFWM) to characterize the real and imaginary parts of Chi((3)) using an imaged 2-D phase grating. Phase sensitive coherent detection of DFWM signal is demonstrated. Phase stabilization of the interferometric arms is obtained passively with the 2-D grating. A processable polyacetylene sample is characterized at a wavelength of 1.5 microm using this technique. The observed nonlinearity is determined to be a fast (<250 fs) effect using a simple test.


Resveratrol and cardiovascular health--promising therapeutic or hopeless illusion?
Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a natural polyphenolic compound that exists in Polygonum cuspidatum, grapes, peanuts and berries, as well as their manufactured products, especially red wine. Resveratrol is a pharmacologically active compound that interacts with multiple targets in a variety of cardiovascular disease models to exert protective effects or induce a reduction in cardiovascular risks parameters. This review attempts to primarily serve to summarize the current research findings regarding the putative cardioprotective effects of resveratrol and the molecular pathways underlying these effects. One intent is to hopefully provide a relatively comprehensive resource for clues that may prompt ideas for additional mechanistic studies which might further elucidate and strengthen the role of the stilbene family of compounds in cardiovascular disease and cardioprotection. Model systems that incorporate a significant functional association with tissues outside of the cardiovascular system proper, such as adipose (cell culture, obesity models) and pancreatic (diabetes) tissues, were reviewed, and the molecular pathways and/or targets related to these models and influenced by resveratrol are discussed. Because the body of work encompassing the stilbenes and other phytochemicals in the context of longevity and the ability to presumably mitigate a plethora of afflictions is replete with conflicting information and controversy, especially so with respect to the human response, we tried to remain as neutral as possible in compiling and presenting the more current data with minimal commentary, permitting the reader free reign to extract the knowledge most helpful to their own investigations.


Growth charts for children with Down syndrome: 1 month to 18 years of age.
Centile charts for assessment of stature and weight reflecting expected deficient size and growth rate of home-reared children with Down syndrome are presented for two age intervals, 1 to 36 months and 2 to 18 years, based on 4650 observations on 730 children. Data were pooled and used to estimate five centiles which were smoothed using a flexible mathematical function. These data corroborate other studies of growth in children with Down syndrome demonstrating deficient growth rate throughout the growing period, but most marked in infancy and again at adolescence. Children with Down syndrome in the present sample were taller than those from institutionalized samples at all ages throughout the growing period. Children with moderate or severe congenital heart disease on average were 1.5 to 2.0 cm shorter and about 1 kg lighter than those without or with only mild disease. Mean weight and weight divided by stature squared show that children with Down syndrome have a tendency to be overweight beginning in late infancy and throughout the remainder of the growing years.


RIP1-dependent Bid cleavage mediates TNFα-induced but Caspase-3-independent cell death in L929 fibroblastoma cells.
L929 fibroblastoma cells (L929-A) and L929 fibrosarcoma cells (L929-N) are different cell lines that are commonly used to study the cytotoxicity of tumor necrosis factor alpha (TNFα). TNFα has been reported to induce necrosis in both of these cell lines. However, comparing the TNFα-induced cell death in these two cell lines, we found that, unlike the L929-N cells that show typical RIP3-dependent necrosis, TNFα-induced cell death in L929-A cells is pan-caspase inhibitor Z-VAD-FMK (Z-VAD)-sensitive, which does not depend on RIP3. We also confirmed that the cell death signal in the L929-A cells was initiated through cytosol-preassembled ripoptosome and that the knockdown of either Caspase-8 or RIP1 protein blocked cell death. Compared with the L929-N cells, the L929-A cell line had lower levels of constitutive and inducible TNFα autocrine production, and the pan-caspase inhibitors Z-VAD or Q-VD did not kill the L929-A cells as they affect the L929-N cells. Moreover, the L929-A cells expressed less RIP3 protein than the L929-N cells; therefore, TNFα failed to induce RIP3-dependent necroptosis. In addition, the ripoptosome-mediated cell death signal was transduced to the mitochondria through Caspase-8-mediated and RIP1 kinase activity-dependent Bid cleavage. The RIP1 kinase inhibitor Necrostatin-1 (Nec-1) or Caspase-8 knockdown completely blocked Bid cleavage, and the knockdown of Bid or Bax/Bak prevented TNFα-induced cell death in the L929-A cells. Although the activation of Bax/Bak decreased the mitochondrial membrane potential, the levels of mitochondrial intermembrane space proteins, including cytochrome-c (cyt-C) and Smac, declined, and western blotting and immunofluorescence staining analysis did not determine whether these proteins were redistributed to the cytosol. In addition, the mitochondrial outer membrane protein Tom20 was also reduced, indicating that the reduced mitochondria proteins may be induced by the reduced mitochondria numbers. No efficient cyt-C release was observed; therefore, the limited activation and cleavage of downstream caspases, including Caspase-9, Caspase-7, Caspase-6 and Caspase-3, was insufficient to kill the cells. The Caspase-9, Caspase-6 and Caspase-3/7 inhibitors or Caspase-9 and -3 knockdown also failed to block cell death, and the overexpression of Bcl-2 also did not abrogate cell death. Moreover, the dead cells showed necrotic-like but not apoptotic characteristics under transmission electronmicroscopy, and these features were significantly different from mitochondrial apoptosis, indicating that the effector caspases were not the executioners of cell death. These new discoveries show that TNFα-induced cell death in L929-A cells is different than typical RIP3-dependent necrosis and Caspase-8/Caspase-3-mediated apoptosis. These results highlight that caution is necessary when using different L929 cells as a model to investigate TNFα-induced cell death.


Effect of Morphology and Size of Halloysite Nanotubes on Functional Pectin Bionanocomposites for Food Packaging Applications.
Pectin bionanocomposite films filled with various concentrations of two different types of halloysite nanotubes were prepared and characterized in this study as potential films for food packaging applications. The two types of halloysite nanotubes were long and thin (patch) (200-30 000 nm length) and short and stubby (Matauri Bay) (50-3000 nm length) with different morphological, physical, and dispersibility properties. Both matrix (pectin) and reinforcer (halloysite nanotubes) used in this study are considered as biocompatible, natural, and low-cost materials. Various characterization tests including Fourier transform infrared spectroscopy, field emission scanning electron microscopy, release kinetics, contact angle, and dynamic mechanical analysis were performed to evaluate the performance of the pectin films. Exceptional thermal, tensile, and contact angle properties have been achieved for films reinforced by patch halloysite nanotubes due to the patchy and lengthy nature of these tubes, which form a bird nest structure in the pectin matrix. Matauri Bay halloysite nanotubes were dispersed uniformly and individually in the matrix in low and even high halloysite nanotube concentrations. Furthermore, salicylic acid as a biocidal agent was encapsulated in the halloysite nanotubes lumen to control its release kinetics. On this basis, halloysite nanotubes/salicylic acid hybrids were dispersed into the pectin matrix to develop functional biofilms with antimicrobial properties that can be extended over time. Results revealed that shorter nanotubes (Matauri Bay) had better ability for the encapsulation of salicylic acid into their lumen, while patchy structure and longer tubes of patch halloysite nanotubes made the encapsulation process more difficult, as they might need more time and energy to be fully loaded by salicylic acid. Moreover, antimicrobial activity of the films against four different strains of Gram-positive and Gram-negative bacteria indicated the effective antimicrobial properties of pectin/halloysite functionalized films and their potential to be used for food packaging applications.


Magnetic resonance imaging and intraocular foreign bodies.
We conducted in vitro and in vivo experiments to study magnetic resonance imaging of intraocular foreign bodies. Diamagnetic and paramagnetic foreign bodies were imaged without artifact and without movement during the imaging process, while ferromagnetic foreign bodies produced large amounts of artifact that prevented meaningful images. All fetromagnetic foreign bodies moved during in vitro imaging. During in vivo imaging, three of four ferromagnetic bodies moved, producing substantial retinal injury. We concluded that magnetic resonance imaging is contraindicated in traumatized eyes with suspected ferromagnetic foreign bodies.


Commiphora myrrha Resin Alcoholic Extract Ameliorates High Fat Diet Induced Obesity via Regulation of UCP1 and Adiponectin Proteins Expression in Rats.
This study was performed to evaluate anti-obesity potential of Commiphora myrrha resin ethanolic extract (CME) with the respect to expression of leptin, adiponectin and uncoupling protein 1 (UCP1) in rats. Control rats fed basal diet. Second group fed basal diet and administered CME (500 mg/kg bw) orally for 14 weeks. Third group fed high fat diet (HFD) for 14 weeks. Fourth group fed HFD and administered CME as second group. Fifth group fed HFD for 8 weeks then fed basal diet and administered CME as third group for another 6 weeks. Phytochemical analysis of CME identified the presence of germacrene B, 1,4-benzoquinone, benzofuran, hexadecanoic acid, 9,12-octadecnoic acid methyl ester, reynosin, 11, 14-eicosadienoic acid, isochiapin B, bisabolene epixod, elemene and 1-heptatriacotanol. High fat diet significantly increased food intake, body weight, hyperglycemia, serum levels of total cholesterol, triacylglycerol, low density lipoprotein and ketone bodies, AST and AST activities, concentration of malondialdehyde and histopathological changes in hepatic tissues. However, it significantly reduced serum levels of high density lipoprotein, leptin and adiponectin, activity of hepatic glutathione reductase (GR) and brown adipose tissue UCP1 protein expression. In contrast, CME ameliorated HFD increased body weight, hyperglycemia, dyslipidemia, ketonemia, hepatic tissues lipid peroxidation, restored hepatic tissue architecture and enhanced protein expression of leptin, adiponectin and UCP1 and activity of hepatic GR. This study indicated that CME ameliorated HFD induced hyperglycemia and dyslipidemia through normalization of HFD reduced leptin, adiponectin and UCP1 proteins production and antioxidant activity.


Yeast importin-β is required for nuclear import of the Mig2 repressor.
Mig2 has been described as a transcriptional factor that in the absence of Mig1 protein is required for glucose repression of the SUC2 gene. Recently it has been reported that Mig2 has two different subcellular localizations. In high-glucose conditions it is a nuclear modulator of several Mig1-regulated genes, but in low-glucose most of the Mig2 protein accumulates in mitochondria. Thus, the Mig2 protein enters and leaves the nucleus in a glucose regulated manner. However, the mechanism by which Mig2 enters into the nucleus was unknown until now. Here, we report that the Mig2 protein is an import substrate of the carrier Kap95 (importin-β). The Mig2 nuclear import mechanism bypasses the requirement for Kap60 (importin-α) as an adaptor protein, since Mig2 directly binds to Kap95 in the presence of Gsp1(GDP). We also show that the Mig2 nuclear import and the binding of Mig2 with Kap95 are not glucose-dependent processes and require a basic NLS motif, located between lysine-32 and arginine-37. Mig2 interaction with Kap95 was assessed in vitro using purified proteins, demonstrating that importin-β, together with the GTP-binding protein Gsp1, is able to mediate efficient Mig2-Kap95 interaction in the absence of the importin-α (Kap60). It was also demonstrated, that the directionality of Mig2 transport is regulated by association with the small GTPase Gsp1 in the GDP- or GTP-bound forms, which promote cargo recognition and release, respectively. The Mig2 protein accumulates in the nucleus through a Kap95 and NLS-dependent nuclear import pathway, which is independent of importin-α in Saccharomyces cerevisiae.


Climate-induced variations in global wildfire danger from 1979 to 2013.
Climate strongly influences global wildfire activity, and recent wildfire surges may signal fire weather-induced pyrogeographic shifts. Here we use three daily global climate data sets and three fire danger indices to develop a simple annual metric of fire weather season length, and map spatio-temporal trends from 1979 to 2013. We show that fire weather seasons have lengthened across 29.6 million km(2) (25.3%) of the Earth's vegetated surface, resulting in an 18.7% increase in global mean fire weather season length. We also show a doubling (108.1% increase) of global burnable area affected by long fire weather seasons (>1.0 σ above the historical mean) and an increased global frequency of long fire weather seasons across 62.4 million km(2) (53.4%) during the second half of the study period. If these fire weather changes are coupled with ignition sources and available fuel, they could markedly impact global ecosystems, societies, economies and climate.


Positive clinical outcomes derived from using a proprietary mixture of selected strains during pregnancy.
While the use of antibiotic prophylaxis is clearly advantageous to prevent streptococcal infection, it alters the composition of the gut microbiota in mothers and infants. Enterococcus faecium L3 is one of the best studied probiotic strains and shows strong antagonistic activity against Streptococcus agalactiae due to the production of bacteriocins able to inhibit common gut and vaginal pathogens. We tested an L3-based probiotic formula (iNatal®) on 127 pregnant women attending our gynaecological unit in 2015. We compared the study subjects with 279 pregnant women enrolled in the same year and with 892 other pregnant women who attended our gynaecological unit in 2013 and 2014. The findings demonstrate: (a) the safety profile of the product; (b) its ability to reduce gut disorders; (c) a 6% decrease in the incidence of streptococcal colonization; (d) an approximately 30% decline in episodes of premature rupture of membranes; (e) fewer caesarean sections during labour; and (f) a reduction in pathological umbilical cord blood pH. Our results demonstrate that a probiotic treatment during pregnancy could have unexpected but favourable clinical results. Further randomized, double-blind, placebo controlled studies are now needed to confirm our preliminary findings.


Prolonged psychosis after Amanita muscaria ingestion.
Amanita muscaria has a bright red or orange cap covered with small white plaques. It contains the isoxazole derivatives ibotenic acid, muscimol and muscazone and other toxins such as muscarine. The duration of clinical manifestations after A. muscaria ingestion does not usually exceed 24 hours; we report on a 5-day paranoid psychosis after A. muscaria ingestion. A 48-year-old man, with no previous medical history, gathered and ate mushrooms he presumed to be A. caesarea. Half an hour later he started to vomit and fell asleep. He was found comatose having a seizure-like episode. On admission four hours after ingestion he was comatose, but the remaining physical and neurological examinations were unremarkable. Creatine kinase was 8.33 microkat/l. Other laboratory results and brain CT scan were normal. Toxicology analysis did not find any drugs in his blood or urine. The mycologist identified A. muscaria among the remaining mushrooms. The patient was given activated charcoal. Ten hours after ingestion, he awoke and was completely orientated; 18 hours after ingestion his condition deteriorated again and he became confused and uncooperative. Afterwards paranoid psychosis with visual and auditory hallucinations appeared and persisted for five days. On the sixth day all symptoms of psychosis gradually disappeared. One year later he is not undergoing any therapy and has no symptoms of psychiatric disease. We conclude that paranoid psychosis with visual and auditory hallucinations can appear 18 hours after ingestion of A. muscaria and can last for up to five days.


Selective Granulocyte and Monocyte Apheresis as a Non-Pharmacological Option for Patients with Inflammatory Bowel Disease.
Ulcerative colitis and Crohn's disease are the two most prevalent inflammatory bowel diseases. In both cases, the medically refractory and steroid-dependent type presents a therapeutic challenge. To help resolve this problem, a mainly Japanese team developed a new therapeutic option. There are two systems, both of which are able to selectively remove the main mediators of the disease, namely the activated pro-inflammatory cytokine-producing granulocytes and monocytes/macrophages, from the patient's blood circulation (GMA = granulocyte monocyte apheresis). One of the two systems is the Adacolumn( (®) ) (Immunoresearch Laboratories, Takasaki, Japan) consisting of the ADA-monitor and a single-use column, which contains approximately 35,000 cellulose acetate beads. The exact mode of action is not yet sufficiently understood, but however, a modulation of the immune system takes place. As a result, less pro-inflammatory cytokines are released. Furthermore, the production of anti-inflammatory interleukin-1 receptor antagonist is increased, and the apoptosis of granulocytes boosted. The decreased LECAM-1-expression on leukocytes impedes the leukotaxis to the inflamed tissue, and CD10-negative immature granulocytes appear in the peripheral blood. Another effect to be mentioned is the removal of the peripheral dendritic cells and the leachate of regulatory T cells (T-regs). The second system is the Cellsorba( (®) ) FX Filter (Asahi Medical, Tokyo, Japan). The range of efficiency, the indication, and the procedure are very similar to the Adacolumn. Solely the additional removal of lymphocytes can possibly limit the implementation since lymphopenia can increase the risk of autoimmune disease. Both systems provide a low-risk therapy with few adverse reactions. ASFA recommendations for GMA in inflammatory bowel disease are 2B due to the fact that not enough randomized double-blind studies are available to proof the efficacy of this treatment.


A Comparison of Tapered and Nontapered Implants in the Minipig.
Tapered implants are thought to result in equivalent long-term stability and marginal peri-implant bone height in comparison to cylindrical implants. The goal of this study was to compare the primary stability and osseointegration of a novel bone-level tapered-wall implant to a control bone-level cylindrical implant with exactly the same surface characteristics, in a direct side-by-side comparison in an animal model. Mandibular premolars and first molars were extracted in 11 G.ttingen minipigs, and healing was allowed for 3 months. Six implants were placed with a 4-week surgical lag time between hemimandibles (three per side), and either immediately loaded (first implantation time point) or submerged (second implantation time point). Eight-mm-long × 4.1-mm-diameter titanium screw-type bone-level implants with tapered (two per side) and cylindrical bodies (one per side) were used (Institut Straumann); insertion torque and implant stability were measured, and the soft tissue was closed. Standardized radiographs were taken at implant placement and at sacrifice. Following a healing period of 1 month from the last surgical intervention (8 weeks total healing time for loaded implants; 4 weeks total healing time for nonloaded implants), the animals were sacrificed and mandibular blocks were harvested for nondecalcified histologic and histomorphometric analysis. All implants were integrated radiographically and osseointegrated histologically. Maximum insertion torque measurements and implant stability quotient values showed no significant difference between tapered and cylindrical implants. Histomorphometric analysis also resulted in comparable bone-to-implant contact values between the implant types and similarly limited marginal peri-implant bone resorption; no significant differences were observed regarding all the evaluated parameters between the groups, regardless of the loading regime. In a direct side-by-side comparison, in an intraoral minipig model, a novel bone-level tapered screw implant with an SLActive surface showed comparable clinical and histologic outcomes to a parallel-walled bone-level screw implant with an SLActive surface.


Multi-modal two-step floating catchment area analysis of primary health care accessibility.
Two-step floating catchment area (2SFCA) techniques are popular for measuring potential geographical accessibility to health care services. This paper proposes methodological enhancements to increase the sophistication of the 2SFCA methodology by incorporating both public and private transport modes using dedicated network datasets. The proposed model yields separate accessibility scores for each modal group at each demand point to better reflect the differential accessibility levels experienced by each cohort. An empirical study of primary health care facilities in South Wales, UK, is used to illustrate the approach. Outcomes suggest the bus-riding cohort of each census tract experience much lower accessibility levels than those estimated by an undifferentiated (car-only) model. Car drivers' accessibility may also be misrepresented in an undifferentiated model because they potentially profit from the lower demand placed upon service provision points by bus riders. The ability to specify independent catchment sizes for each cohort in the multi-modal model allows aspects of preparedness to travel to be investigated.


Culture and differentiation of rat neural stem/progenitor cells in a three-dimensional collagen scaffold.
A stable and fast method for constructing a neural-like tissue from rat neural stem/progenitor cells (rNS/PCs) based on three-dimensional (3D) collagen gel is described. First step, the collagen-embedded rNS/PCs expanded with the medium consisting of DMEM/F12/RPMI1640 (1:1:1) supplemented with EGF and bFGF was used to expand the cells in gel in 96-well plates until the average diameter of cell clusters was about 50-100 μm with the cell density higher than 10(7) cells/mL. In the second step, the initial medium was replaced with NB/B-27 supplemented with bFGF and BDNF. The results show that cells in collagen presented neural-like morphology and maintained live cell rate around 82 % in neural network pattern at least for 42 days under static conditions. The cell-collagen constructs were detected by immunofluorescence and immunohistochemistry test after 42 days of culture, part of cells still maintained the character of rNS/PCs, and others differentiated into neurons, astrocytes, and oligodendrocytes. Our 3D neural-like tissue construct was similar to the neural tissue in morphology and cell compositions. They thus have a potential to be used for drug screening, detection of environment toxins, and replacement therapy.