A faster and simpler UPLC-MS/MS method for the simultaneous determination of trimethylamine N-oxide, trimethylamine and dimethylamine in different types of biological samples.
Gut microbiota-dependent metabolites trimethylamine N-oxide (TMAO), trimethylamine (TMA) and dimethylamine (DMA) from dietary methylamines have recently gained much attention due to their high association with chronic kidney disease risk. Hence a simpler and faster performance liquid chromatography-tandem mass spectrometry method was developed and validated. The quantitative analysis was achieved within 6 min by using Agilent 6460C UPLC-MS/MS with 10% methyl alcohol isocratic elution and was more simple, convenient and rapid than that of previously reported methods. Furthermore, method verification results showed that the method correlation coefficient was 0.99978293, 0.99997514 and 0.98784721, and the detection limit was 0.121, 8.063 and 0.797 μg L-1, and the precision of the retention time and peak area of analytes was less than 0.331 and 3.280, respectively. The method was applied to simultaneously determine TMAO, TMA and DMA in the urine and serum from mice treated with normal, high l-carnitine, or high choline diet. Quantitative recoveries of TMAO, TMA and DMA were in the range of 94.2%-101.0%, and the RSD values were lower than 5.17%. The proposed UPLC-MS/MS-based assay should be of value for further evaluating TMAO as a risk marker and for examining the effect of dietary factors on TMAO metabolism.


Clinical predictors of chronic renal allograft rejection.
Among both immune and nonimmune factors implicated in the pathogenesis of chronic renal allograft rejection, acute rejection episodes represent a strong and consistent predictor. However, all acute rejection episodes are not equally predictive of chronic renal allograft rejection. Early and mild acute rejection episodes do not usually cause chronic renal allograft rejection. On the other hand, late and severe acute rejection episodes occurring more than one year after transplantation are particularly strong predictors of chronic renal allograft rejection. The number of HLA mismatches is a risk factor, but its influence may not be independent of acute rejection and other risk factors. Proteinuria and recently hypoalbuminemia are also independent risk factors for chronic renal allograft rejection. However, whether these nonimmune factors are merely the result of chronic renal allograft rejection, or whether they contribute to the pathogenesis of renal injury in chronic renal allograft rejection is yet unclear. Better HLA matching, new strategies to decrease the severity of acute rejection, and measures to prevent late acute rejection, as well as prospective evaluation of the therapies to reduce proteinuria and other nonimmune risk factors for chronic renal allograft rejection are needed.


Six Degree-of-Freedom Haptic Simulation of a Stringed Musical Instrument for Triggering Sounds.
Six degree-of-freedom (DoF) haptic rendering of multi-region contacts between a moving hand avatar and varied-shaped components of a music instrument is fundamental to realizing interactive simulation of music playing. There are two aspects of computational challenges: first, some components have significantly small sizes in some dimensions, such as the strings on a seven-string plucked instrument (e.g., Guqin), which makes it challenging to avoid pop-through during multi-region contact scenarios. Second, deformable strings may produce high-frequency vibration, which requires simulating diversified and subtle force sensations when a hand interacts with strings in different ways. In this paper, we propose a constraint-based approach to haptic interaction and simulation between a moving hand avatar and various parts of a string instrument, using a cylinder model for the string that has a large length-radius ratio and a sphere-tree model for the other parts that have complex shapes. Collision response algorithms based on configuration-based optimization is adapted to solve for the contact configuration of the hand avatar interacting with thin strings without penetration. To simulate the deformation and vibration of a string, a cylindrical volume with variable diameters is defined with response to the interaction force applied by the operator. Experimental results have validated the stability and efficiency of the proposed approach. Subtle force feelings can be simulated to reflect varied interaction patterns, to differentiate collisions between the hand avatar with a static or vibrating string and the effects of various colliding forces and touch locations on the strings.


Epidemiology of community-associated Clostridium difficile infection, 2009 through 2011.
Clostridium difficile infection (CDI) has been increasingly reported among healthy individuals in the community. Recent data suggest that community-associated CDI represents one-third of all C difficile cases. The epidemiology and potential sources of C difficile in the community are not fully understood. To determine epidemiological and clinical characteristics of community-associated CDI and to explore potential sources of C difficile acquisition in the community. Active population-based and laboratory-based CDI surveillance in 8 US states. Medical records were reviewed and interviews performed to assess outpatient, household, and food exposures among patients with community-associated CDI (ie, toxin or molecular assay positive for C difficile and no overnight stay in a health care facility within 12 weeks). Molecular characterization of C difficile isolates was performed. Outpatient health care exposure in the prior 12 weeks among patients with community-associated CDI was a priori categorized into the following 3 levels: no exposure, low-level exposure (ie, outpatient visit with physician or dentist), or high-level exposure (ie, surgery, dialysis, emergency or urgent care visit, inpatient care with no overnight stay, or health care personnel with direct patient care). Prevalence of outpatient health care exposure among patients with community-associated CDI and identification of potential sources of C difficile by level of outpatient health care exposure. Of 984 patients with community-associated CDI, 353 (35.9%) did not receive antibiotics, 177 (18.0%) had no outpatient health care exposure, and 400 (40.7%) had low-level outpatient health care exposure. Thirty-one percent of patients without antibiotic exposure received proton pump inhibitors. Patients having CDI with no or low-level outpatient health care exposure were more likely to be exposed to infants younger than 1 year (P = .04) and to household members with active CDI (P = .05) compared with those having high-level outpatient health care exposure. No association between food exposure or animal exposure and level of outpatient health care exposure was observed. North American pulsed-field gel electrophoresis (NAP) 1 was the most common (21.7%) strain isolated; NAP7 and NAP8 were uncommon (6.7%). Most patients with community-associated CDI had recent outpatient health care exposure, and up to 36% would not be prevented by reduction of antibiotic use only. Our data support evaluation of additional strategies, including further examination of C difficile transmission in outpatient and household settings and reduction of proton pump inhibitor use.


Modulation of event-related desynchronization during motor imagery with transcranial direct current stimulation (tDCS) in patients with chronic hemiparetic stroke.
Electroencephalogram-based brain-computer interface (BCI) has been developed as a new neurorehabilitative tool for patients with severe hemiparesis. However, its application has been limited because of difficulty detecting stable brain signals from the affected hemisphere. It has been reported that transcranial direct current stimulation (tDCS) can modulate event-related desynchronization (ERD) in healthy persons. The objective of this study was to test the hypothesis that anodal tDCS could modulate ERD in patients with severe hemiparetic stroke. The participants were six patients with chronic hemiparetic stroke (mean age, 56.8 ± 9.5 years; mean time from the onset, 70.0 ± 19.6 months; Fugl-Meyer Assessment upper extremity motor score, 30.8 ± 16.5). We applied anodal tDCS (10 min, 1 mA) and sham stimulation over the affected primary motor cortex in a random order. ERD of the mu rhythm (mu ERD) with motor imagery of extension of the affected finger was assessed before and after anodal tDCS and sham stimulation. Mu ERD of the affected hemisphere increased significantly after anodal tDCS, whereas it did not change after sham stimulation. Our results show that anodal tDCS can increase mu ERD in patients with hemiparetic stroke, indicating that anodal tDCS could be used as a conditioning tool for BCI in stroke patients.


White matter microstructure correlates of inhibition and task-switching in adolescents.
Although protracted prefrontal gray matter development is associated with concomitant executive function (EF) development in adolescents, few studies have explored the relationship between white matter and EF. This study examined the relationship between white matter microstructure and two aspects of EF, inhibition and task-switching, in a sample of 84 adolescents using diffusion tensor imaging (DTI). Tract-Based Spatial Statistics (TBSS) were used to examine fractional anisotropy (FA) and mean diffusivity (MD). Adolescents completed the Color-Word Interference task from the Delis-Kaplan Executive Function System, a clinical version of the Stroop task. Inhibition and task-switching performance were group normalized and measured using both reaction time and errors. Performance and the interaction of age and performance were regressed on FA and MD white matter skeletons, controlling for age and IQ, separately for inhibition and task-switching. Follow up analyses examined the relative contributions of axial and radial diffusivities. Greater FA in the anterior corona radiata (ACR) was associated with better inhibition, independent of age. Greater FA in the SCR and precentral gyrus white matter were associated with better task-switching, regardless of age, whereas an association between FA in the ACR and task-switching was dependent on age. There were no significant associations between MD and performance. Results suggest better inhibition and task-switching are associated with greater integrity of white matter microstructure in regions supporting cross-cortical and cortical-subcortical connections stemming from the prefrontal cortex. These findings are consistent with functional studies of cognitive control and models of EF that propose separate, yet related, latent factors.


[Cardiovascular molecular genetics and gene therapy. Introduction to the terminology and technique].
The possible application of somatic gene therapy to the treatment of cardiovascular diseases is introduced; methods and delivery systems for transferring genes to the sites of application are discussed. Although cardiovascular diseases are mainly systemic disorders, critical lesions are frequently located at specific sites in the circulation and therefore accessible to the introduction of recombinant genes which may encode for proteins either inhibiting cell proliferation or promoting cell division. Cell-mediated gene transfer is compared to direct gene transfer in vivo; the advantages and disadvantages of the "ex vivo-in vitro-in vivo" and "direct in vivo" techniques are described. Several alternative gene transfer techniques are known. They include viral and retroviral vectors which have been made replication defective by genetic engineering, as well as non-viral systems such as DNA-liposome complexes, and physical methods such as the so-called biolistic technology originally developed to penetrate plant cell membranes. In summary, it is pointed out that even demonstrations of the few successful therapeutic applications to date do not conceal the fact that intensive experimental research using animal models is needed to prove the advantages and safety of the techniques used in gene therapy of the cardiovascular system.


Down-regulation of Inositol Polyphosphate 4-Phosphatase Type II Expression in Colorectal Carcinoma.
Aberrant expression of survival signaling pathways causes deregulation of cellular proliferation and resistance to apoptosis, and plays a crucial role in the development, progression and metastasis of cancer. Inositol polyphosphate 4-phosphatase type II (INPP4B) negatively regulates phosphatidylinositol 3-kinase signaling and has a tumor-suppressive role in several human malignancies. We analyzed the expression levels of INPP4B mRNA and protein in colorectal carcinoma (CRC) cell lines and tissue samples using western blot, quantitative real-time reverse-transcriptase polymerase chain reaction, and immunohistochemical staining. Western blot analysis revealed that the CRC cell lines HCT 116, SW620, DLD-1, and WiDr expressed significantly lower levels of INPP4B protein than the normal colonic epithelial cell lines CCD 841 CoTr and FHC. Consistent with these results, INPP4B mRNA expression in the CRC cell lines was significantly lower than in the normal colonic epithelial cells. Immunohistochemical staining revealed that normal colonic mucosa displayed uniform and strong-to-moderate INPP4B immunoreactivity, whereas 60.7% (71/117; p<0.001) and 76.5% (62/81; p<0.001) of the primary and metastatic CRC tissue samples exhibited reduced INPP4B expression, respectively. Our results indicate that INPP4B is down-regulated in CRC and that INPP4B is involved in the development and progression of CRC.


[Hygienic evaluation of ambient air pollution in Ust'-Kamenogorsk].
The article deals with hygienic evaluation of Ust-Kamenogorsk ambient air pollution with nitrogen dioxide, carbon dioxide, phenol and suspended solids. Ambient air samples were taken in 36 points over 3 days at 6 am, noon, 6 and 9 pm (local time) during warm season (August) and cold season (December). According to the results obtained, the authors divided Ust-Kamenogorsk territory into 2 areas, based on IAP4 (index of air pollution): conditionally dirty area--over 7; conditionally clean area--under 7. Findings are high pollution with suspended solids, nitrogen dioxide, sulphur dioxide and phenol over all territory of the city. But "conditionally dirty" area demonstrated the highest level of ambient air pollution.


Pramlintide in the management of insulin-using patients with type 2 and type 1 diabetes.
In patients with diabetes, dysregulation of multiple glucoregulatory hormones results in chronic hyperglycemia and an array of associated microvascular and macrovascular complications. Optimization of glycemic control, both overall (glycosylated hemoglobin [A1C]) and in the postprandial period, may reduce the risk of long-term vascular complications. However, despite significant recent therapeutic advances, most patients with diabetes are unable to attain and/or maintain normal or near-normal glycemia with insulin therapy alone. Pramlintide, an analog of amylin, is the first in a new class of pharmaceutical agents and is indicated as an adjunct to mealtime insulin for the treatment of patients with type 1 and type 2 diabetes. By mimicking the actions of the naturally occurring hormone amylin, pramlintide complements insulin by regulating the appearance of glucose into the circulation after meals via three primary mechanisms of action: slowing gastric emptying, suppressing inappropriate post-meal glucagon secretion, and increasing satiety. In long-term clinical trials, adjunctive pramlintide treatment resulted in improved postprandial glucose control and significantly reduced A1C and body weight compared with insulin alone. The combination of insulin and pramlintide may provide a more physiologically balanced approach to managing diabetes.


Modeling autosomal dominant Alzheimer's disease with machine learning.
Machine learning models were used to discover novel disease trajectories for autosomal dominant Alzheimer's disease. Longitudinal structural magnetic resonance imaging, amyloid positron emission tomography (PET), and fluorodeoxyglucose PET were acquired in 131 mutation carriers and 74 non-carriers from the Dominantly Inherited Alzheimer Network; the groups were matched for age, education, sex, and apolipoprotein ε4 (APOE ε4). A deep neural network was trained to predict disease progression for each modality. Relief algorithms identified the strongest predictors of mutation status. The Relief algorithm identified the caudate, cingulate, and precuneus as the strongest predictors among all modalities. The model yielded accurate results for predicting future Pittsburgh compound B (R2 = 0.95), fluorodeoxyglucose (R2 = 0.93), and atrophy (R2 = 0.95) in mutation carriers compared to non-carriers. Results suggest a sigmoidal trajectory for amyloid, a biphasic response for metabolism, and a gradual decrease in volume, with disease progression primarily in subcortical, middle frontal, and posterior parietal regions.


Effectiveness of Acupuncture in the Treatment of Parkinson's Disease: An Overview of Systematic Reviews.
Background: The effects of acupuncture on Parkinson's disease (PD) outcomes remain unclear. The aim of this overview was to comprehensively evaluate the methodological quality and applicability of the results of systematic reviews (SRs)/meta-analyses (MAs) that examined the use of acupuncture to treat PD. Methods: Eight databases were searched to retrieve SRs/MAs on the use of acupuncture for the treatment of PD. Two reviewers independently screened and extracted the data using the Assessing the Methodological Quality of Systematic Reviews 2 (AMSTAR-2) checklist to evaluate the methodological quality and using the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) criteria to assess the evidence quality of the included reviews. Results: A total of 11 SRs/MAs were included. According to the AMSTAR-2 checklist results, all included SRs/MAs were rated as very-low-quality studies. The GRADE criteria revealed 20 studies with very-low-quality evidence, 9 with low-quality evidence, 3 with moderate-quality evidence, and 0 with high-quality evidence. Descriptive analysis showed that acupuncture appears to be a clinically effective and safe treatment for PD. Conclusions: The use of acupuncture for the treatment of PD may be clinically effective and safe. This conclusion must be interpreted cautiously due to the generally low methodological quality and low quality of evidence of the included studies.


Neural decoding of electrocorticographic signals using dynamic mode decomposition.
Brain-computer interfaces (BCIs) using electrocorticographic (ECoG) signals have been developed to restore the communication function of severely paralyzed patients. However, the limited amount of information derived from ECoG signals hinders their clinical applications. We aimed to develop a method to decode ECoG signals using spatiotemporal patterns characterizing movement types to increase the amount of information gained from these signals. Previous studies have demonstrated that motor information could be decoded using powers of specific frequency bands of the ECoG signals estimated by fast Fourier transform (FFT) or wavelet analysis. However, because FFT is evaluated for each channel, the temporal and spatial patterns among channels are difficult to evaluate. Here, we used dynamic mode decomposition (DMD) to evaluate the spatiotemporal pattern of ECoG signals and evaluated the accuracy of motor decoding with the DMD modes. We used ECoG signals during three types of hand movements, which were recorded from 11 patients implanted with subdural electrodes. From the signals at the time of the movements, the modes and powers were evaluated by DMD and FFT and were decoded using support vector machine. We used the Grassmann kernel to evaluate the distance between modes estimated by DMD (DMD mode). In addition, we decoded the DMD modes, in which the phase components were shuffled, to compare the classification accuracy. The decoding accuracy using DMD modes was significantly better than that using FFT powers. The accuracy significantly decreased when the phases of the DMD mode were shuffled. Among the frequency bands, the DMD mode at approximately 100 Hz demonstrated the highest classification accuracy. DMD successfully captured the spatiotemporal patterns characterizing the movement types and contributed to improving the decoding accuracy. This method can be applied to improve BCIs to help severely paralyzed patients communicate.


Cell adhesion and intracellular calcium signaling in neurons.
Cell adhesion molecules (CAMs) play indispensable roles in the developing and mature brain by regulating neuronal migration and differentiation, neurite outgrowth, axonal fasciculation, synapse formation and synaptic plasticity. CAM-mediated changes in neuronal behavior depend on a number of intracellular signaling cascades including changes in various second messengers, among which CAM-dependent changes in intracellular Ca2+ levels play a prominent role. Ca2+ is an essential secondary intracellular signaling molecule that regulates fundamental cellular functions in various cell types, including neurons. We present a systematic review of the studies reporting changes in intracellular Ca2+ levels in response to activation of the immunoglobulin superfamily CAMs, cadherins and integrins in neurons. We also analyze current experimental evidence on the Ca2+ sources and channels involved in intracellular Ca2+ increases mediated by CAMs of these families, and systematically review the role of the voltage-dependent Ca2+ channels (VDCCs) in neurite outgrowth induced by activation of these CAMs. Molecular mechanisms linking CAMs to VDCCs and intracellular Ca2+ stores in neurons are discussed.


Peak pressures: crossover study of timolol and latanoprost.
To compare the diurnal efficacy and action on peak intraocular pressures (IOP) of 0.005% latanoprost and 0.5% timolol as primary therapy in 60 eyes having dark brown irides with primary open angle glaucoma (POAG). A prospective, comparative, observer-masked, crossover, interventional trial including the mean of both eyes of 30 patients with POAG who were randomly started on either latanoprost once daily or timolol twice daily. Three months after treatment with one drug, the second drug was substituted. A masked observer carried out diurnal assessments of IOP before the start of therapy and at 3 and 7 months. The fourth month was the washout period for the first drug. The average baseline IOP was 23.36 +/- 2.14 mm Hg, which was reduced by 8.8 +/- 2.2 mmHg with latanoprost (p < 0.01) and by 6.75 +/- 1.9 mm Hg with timolol (p = 0.01). The reduction was greater for latanoprost (p < 0.005). The average peak IOP at baseline was 27.6 +/- 2.22 mmHg. The effective fall in IOP at the time of new peaks in subsequent diurnal recordings of IOP compared to the baseline diurnal curve was 8.9 mm Hg with latanoprost (p < 0.005) and 5.77 mm Hg with timolol (p < 0.01). This difference in IOP reduction between the two drugs was statistically significant (p < 0.01). Latanoprost had a lower efficacy in peak IOP reduction in eyes with evening peak of IOP than in those with morning peak (p < 0.005). The efficacy of timolol was lower overall compared to latanoprost, but was similar in all circadian rhythms. The shift in timing of IOP peak was greater with latanoprost compared to timolol (4.34 hours vs -0.72 hours, p < .01). A total of 90% of patients on latanoprost and 33.3% on timolol achieved a reduction of > 30% in baseline mean IOP. The average of the trough IOP recorded in each of the individual baseline IOP curves was 19.05 +/- 2.05 mm Hg. Greater mean and peak IOP reduction was achieved with latanoprost compared to timolol. Dampening of the circadian rhythm was better with latanoprost. Latanoprost appears to be more effective than timolol at all points in time with greater efficacy in eyes with morning peaks compared to evening peaks.


Gender differences in occupational exposure patterns.
The authors conducted a population-based survey to examine gender differences in occupational exposure patterns and to investigate whether any observed differences are due to: (a) gender differences in occupational distribution; and/or (b) gender differences in tasks within occupations. Men and women aged 20-64 years were randomly selected from the Electoral Roll and invited to take part in a telephone interview, which collected information on self-reported occupational exposure to specific dusts and chemicals, physical exposures and organisational factors. The authors used logistic regression to calculate prevalence ORs and 95% CIs comparing the exposure prevalence of males (n=1431) and females (n=1572), adjusting for age. To investigate whether men and women in the same occupation were equally exposed, the authors also matched males to females on current occupation using the five-digit code (n=1208) and conducted conditional logistic regression adjusting for age. Overall, male workers were two to four times more likely to report exposure to dust and chemical substances, loud noise, irregular hours, night shifts and vibrating tools. Women were 30% more likely to report repetitive tasks and working at high speed, and more likely to report exposure to disinfectants, hair dyes and textile dust. When men were compared with women with the same occupation, gender differences were attenuated. However, males remained significantly more likely to report exposure to welding fumes, herbicides, wood dust, solvents, tools that vibrate, irregular hours and night-shift work. Women remained more likely to report repetitive tasks and working at high speed, and in addition were more likely to report awkward or tiring positions compared with men with the same occupation. This population-based study showed substantial differences in occupational exposure patterns between men and women, even within the same occupation. Thus, the influence of gender should not be overlooked in occupational health research.


The anticodon of the maize chloroplast gene for tRNA Leu UAA is split by a large intron.
The maize chloroplast gene encoding tRNA Leu UAA has been sequenced. It contains a 458 base pair intron between the first and second bases of the anticodon. The tRNA is 88 nucleotides long (the 3'-terminal CCA sequence included which, however, is not encoded by the gene) and differs in only four nucleotides (modified nucleotides are not considered) from the corresponding isoacceptor from bean chloroplasts. The unusual position of the intron in this maize chloroplast tRNA gene suggests a splicing model different from that generally accepted for eukaryotic split tRNA genes.


Tbr1 regulates regional and laminar identity of postmitotic neurons in developing neocortex.
Areas and layers of the cerebral cortex are specified by genetic programs that are initiated in progenitor cells and then, implemented in postmitotic neurons. Here, we report that Tbr1, a transcription factor expressed in postmitotic projection neurons, exerts positive and negative control over both regional (areal) and laminar identity. Tbr1 null mice exhibited profound defects of frontal cortex and layer 6 differentiation, as indicated by down-regulation of gene-expression markers such as Bcl6 and Cdh9. Conversely, genes that implement caudal cortex and layer 5 identity, such as Bhlhb5 and Fezf2, were up-regulated in Tbr1 mutants. Tbr1 implements frontal identity in part by direct promoter binding and activation of Auts2, a frontal cortex gene implicated in autism. Tbr1 regulates laminar identity in part by downstream activation or maintenance of Sox5, an important transcription factor controlling neuronal migration and corticofugal axon projections. Similar to Sox5 mutants, Tbr1 mutants exhibit ectopic axon projections to the hypothalamus and cerebral peduncle. Together, our findings show that Tbr1 coordinately regulates regional and laminar identity of postmitotic cortical neurons.


Transcriptional profiling of whole blood identifies a unique 5-gene signature for myelofibrosis and imminent myelofibrosis transformation.
Identifying a distinct gene signature for myelofibrosis may yield novel information of the genes, which are responsible for progression of essential thrombocythemia and polycythemia vera towards myelofibrosis. We aimed at identifying a simple gene signature - composed of a few genes - which were selectively and highly deregulated in myelofibrosis patients. Gene expression microarray studies have been performed on whole blood from 69 patients with myeloproliferative neoplasms. Amongst the top-20 of the most upregulated genes in PMF compared to controls, we identified 5 genes (DEFA4, ELA2, OLFM4, CTSG, and AZU1), which were highly significantly deregulated in PMF only. None of these genes were significantly regulated in ET and PV patients. However, hierarchical cluster analysis showed that these genes were also highly expressed in a subset of patients with ET (n = 1) and PV (n = 4) transforming towards myelofibrosis and/or being featured by an aggressive phenotype. We have identified a simple 5-gene signature, which is uniquely and highly significantly deregulated in patients in transitional stages of ET and PV towards myelofibrosis and in patients with PMF only. Some of these genes are considered to be responsible for the derangement of bone marrow stroma in myelofibrosis. Accordingly, this gene-signature may reflect key processes in the pathogenesis and pathophysiology of myelofibrosis development.


When, how, and why of advance directives: tools for the cardiovascular nurse.
In the U.S., cardiovascular disease is the number one cause of death. The chronicity and unpredictable nature of cardiovascular disease will affect thousands of patients and families who are often called upon to make difficult end of life choices. The federal legislation of the Patient Self Determination Act (PSDA) was passed in 1990 and went into affect in December of 1991. The essence of this legislation was to empower the public with the right to make end of life decisions. However, after almost 10 years of implementation there is little evidence of national acceptance by the health care industry as evident in the literature. Likewise, the public demonstrates limited knowledge regarding advance directives. The law applies to all health care facilities that receive any federal reimbursement for services and includes hospitals, nursing homes, home health agencies, and clinics. The PSDA requires health care institutions to establish mechanisms for disseminating information about advance directive, to patients and to designate personnel responsible for discussing advance directives with patients. In this paper, practical and clinical application to the special considerations of the patient and end of life choices will be discussed. A focus on facts, resources, and other information will be presented for the cardiovascular nurse in a variety of health care settings.


Molecular recognition of chymotrypsin by the serine protease inhibitor ecotin from Yersinia pestis.
Resistance to antibiotics is a problem not only in terms of healthcare but also biodefense. Engineering of resistance into a human pathogen could create an untreatable biothreat pathogen. One such pathogen is Yersinia pestis, the causative agent of plague. Previously, we have used a bioinformatic approach to identify proteins that may be suitable targets for antimicrobial therapy and in particular for the treatment of plague. The serine protease inhibitor ecotin was identified as one such target. We have carried out mutational analyses in the closely related Yersinia pseudotuberculosis, validating that the ecotin gene is a virulence-associated gene in this bacterium. Y. pestis ecotin inhibits chymotrypsin. Here, we present the structure of ecotin in complex with chymotrypsin to 2.74 Å resolution. The structure features a biologically relevant tetramer whereby an ecotin dimer binds to two chymotrypsin molecules, similar to what was observed in related serine protease inhibitor structures. However, the vast majority of the interactions in the present structure are distinctive, indicating that the broad specificity of the inhibitor for these proteases is based largely on its capacity to recognize features unique to each of them. These findings will have implications for the development of small ecotin inhibitors for therapeutic use.


(99m)Tc-radiolabelled peptides for tumour imaging: present and future.
Receptor-binding peptides have attracted an enormous interest in targeting molecules for the development of tumour specific radiopharmaceutical compounds. The overexpression of many receptors on human tumour makes such peptide-ligands attractive agents for diagnostic imaging and therapy of cancers. The use of solid-phase peptide synthesis and the availability of a wide range of bifunctional chelating agents for the radiolabelling of bioactive peptides with radionuclides have produced a wide variety of useful radiopharmaceutical molecules. For diagnostic purposes, techenetium - 99m is the ideal radionuclide thanks to its nuclear properties and the availability of a low cost portable generator system.


Metformin Hydrochloride Loaded Mucoadhesive Microspheres and Nanoparticles for Anti-Hyperglycemic and Anticancer Effects Using Factorial Experimental Design.
Metformin hydrochloride (HCl) microspheres and nanoparticles were formulated to enhance bioavailability and minimize side effects through sustained action and optimized drug-release characteristics. Initially, the same formulation design with different ratios of metformin HCl and Eudragit RSPO was used to formulate four batches of microspheres and nanoparticles using solvent evaporation and nanoprecipitation methods, respectively. The produced formulations were evaluated based on particle size and shape (particle size distribution (PSD), scanning electron microscope (SEM)), incompatibility (differential scanning calorimetry (DSC), Fourier-transform infrared (FTIR)), drug release pattern, permeation behavior, in vivo hypoglycemic effects, and in vitro anticancer potential. Compatibility studies concluded that there was minimal interaction between metformin HCl and the polymer, whereas SEM images revealed smoother, more spherical nanoparticles than microspheres. Drug release from the formulations was primarily controlled by the non-Fickian diffusion process, except for A1 and A4 by Fickian, and B3 by Super case II. Korsmeyer-Peppas was the best-fit model for the maximum formulations. The best formulations of microspheres and nanoparticles, based on greater drug release, drug entrapment, and compatibility characteristics, were attributed to the study of drug permeation by non-everted intestinal sacs, in vivo anti-hyperglycemic activity, and in vitro anticancer activity. This study suggests that the proposed metformin HCl formulation can dramatically reduce hyperglycemic conditions and may also have anticancer potential.


New Approaches for Absolute Quantification of Stable-Isotope-Labeled Peptide Standards for Targeted Proteomics Based on a UV Active Tag.
Targeted proteomics depends on the availability of stable isotope labeled (SIL) peptide standards, which for absolute protein quantification need to be absolutely quantified. In the present study, three new approaches for absolute quantification of SIL peptides are developed. All approaches rely on a quantification tag (Qtag) with a specific UV absorption. The Qtag is attached to the peptide during synthesis and is removed by tryptic digestion under standard proteomics workflow conditions. While one quantification method (method A) is designed to allow the fast and economic production of absolutely quantified SIL peptides, two other methods (methods B and C) are developed to enable the straightforward re-quantification of SIL peptides after reconstitution to control and monitor known problems related to peptide solubility, precipitation, and adhesion to vials. All methods yield consistent results when compared to each other and when compared to quantification by amino acid analysis. The precise quantitation methods are used to characterize the in vivo specificity of the H3 specific histone methyltransferase EZH2.


[Acquired paraneoplastic hypertrichosis lanuginosa associated with scleroderma].
Acquired hypertrichosis lanuginosa is universally recognized as an individual disease and seldom reported as a genuine paraneoplastic manifestation. We report the case of a 30-year old woman with acquired hypertrichosis lanuginosa. Due to the finding of a cervical lymph node metastasis, she was investigated for an internal neoplasm, but the original tumour could not be found by the usual methods. A bronchogenic carcinoma was discovered at autopsy. Beside hypertrichosis, this patient had other disorders not described in the literature as associated with that disease, viz.: progressive systemic scleroderma, fissured and hyperpigmented tongue, thrombocytopenia, galactorrhoea, axillary and pubic alopecia and overcurvature of toe nails. A review of similar cases in the literature provided clinical arguments in favour of the hormonal origin of this paraneoplastic hypertrichosis.


Antibodies to Herpesvirus hominis types 1 and 2 in malnourished Nigerian children.
Antibodies to Herpesvirus hominis (HVH) types 1 and 2 were determined by a micro-neutralisation method in 37 children with kwashiorkor, 16 with marasmus, and in 64 well-nourished control children. All the children were aged between 1 and 4 years. The prevalence of antibodies was similar in the two sexes and at different ages. HVH-1 antibodies were present in 51% of children with kwashiorkor, in 44% with marasmus, and in 26% of well-nourished children, reflecting the very poor socioeconomic conditions of malnourished children. HVH-2 antibodies too were present in about 19% of children with kwashiorkor, and in 2% of well-nourished controls; they were absent in marasmic children. It is suggested that HVH-2 infection in malnourished children is facilitated by the communal use of fomites--such as bedclothes and underwear.


Ameliorative effect of resveratrol against fluoride-induced alteration of thyroid function in male wistar rats.
Resveratrol (3,4,5-trihydroxystilbene), a polyphenol and well-known natural antioxidant has been evaluated for its protective effect against fluoride-induced metabolic dysfunctions in rat thyroid gland. Fluoride, the most abundant anions present in groundwater throughout the world, creates a major problem in safe drinking water and causes metabolic, structural, and functional injuries in different organ systems. Sub-acute exposure to sodium fluoride at a dose of 20 mg/kg b.w./day orally to rat for 30 days induces thyroidal dysfunction including suppressed synthetic machinery of the thyroid gland to produce nucleic acids and thyroid hormones, mainly T3 and T4. Other functional changes are alteration of certain metabolic enzyme activities like Na(+)-K(+)-ATPase, thyroid peroxidase, and 5,5'-deiodinase. Structural abnormality of thyroid follicles by fluoride intoxication clearly indicates its thyrotoxic manifestation. Resveratrol supplementation in fluoride-exposed animals appreciably prevented metabolic toxicity caused by fluoride and restored both functional status and ultra-structural organization of the thyroid gland towards normalcy. This study first establishes the therapeutic efficacy of resveratrol as a natural antioxidant in thyroprotection against toxic insult caused by fluoride.


Differential sensory fMRI signatures in autism and schizophrenia: Analysis of amplitude and trial-to-trial variability.
Autism and schizophrenia share multiple phenotypic and genotypic markers, and there is ongoing debate regarding the relationship of these two disorders. To examine whether cortical dynamics are similar across these disorders, we directly compared fMRI responses to visual, somatosensory and auditory stimuli in adults with autism (N=15), with schizophrenia (N=15), and matched controls (N=15). All participants completed a one-back letter detection task presented at fixation (to control attention) while task-irrelevant sensory stimulation was delivered to the different modalities. We focused specifically on the response amplitudes and the variability in sensory fMRI responses of the two groups, given the evidence of greater trial-to-trial variability in adults with autism. Both autism and schizophrenia individuals showed weaker signal-to-noise ratios (SNR) in sensory-evoked responses compared to controls (d>0.42), but for different reasons. For the autism group, the fMRI response amplitudes were indistinguishable from controls but were more variable trial-to-trial (d=0.47). For the schizophrenia group, response amplitudes were smaller compared to autism (d=0.44) and control groups (d=0.74), but were not significantly more variable (d<0.29). These differential group profiles suggest (1) that greater trial-to-trial variability in cortical responses may be specific to autism and is not a defining characteristic of schizophrenia, and (2) that blunted response amplitudes may be characteristic of schizophrenia. The relationship between the amplitude and the variability of cortical activity might serve as a specific signature differentiating these neurodevelopmental disorders. Identifying the neural basis of these responses and their relationship to the underlying genetic bases may substantially enlighten the understanding of both disorders.


How to estimate kinship.
The concept of kinship permeates many domains of fundamental and applied biology ranging from social evolution to conservation science to quantitative and human genetics. Until recently, pedigrees were the gold standard to infer kinship, but the advent of next-generation sequencing and the availability of dense genetic markers in many species make it a good time to (re)evaluate the usefulness of genetic markers in this context. Using three published data sets where both pedigrees and markers are available, we evaluate two common and a new genetic estimator of kinship. We show discrepancies between pedigree values and marker estimates of kinship and explore via simulations the possible reasons for these. We find these discrepancies are attributable to two main sources: pedigree errors and heterogeneity in the origin of founders. We also show that our new marker-based kinship estimator has very good statistical properties and behaviour and is particularly well suited for situations where the source population is of small size, as will often be the case in conservation biology, and where high levels of kinship are expected, as is typical in social evolution studies.


Similarities and dissimilarities between the movement ABC-2 and the Zurich neuromotor assessment in children with suspected developmental coordination disorder.
An established tool for the assessment of motor performance in children with developmental coordination disorder (DCD) is the Movement-ABC-2 (M-ABC-2). The Zurich Neuromotor Assessment (ZNA) is also widely used for the evaluation of children's motor performance, but has not been compared with the M-ABC-2. Fifty-one children (39 males) between 5 and 7 years of age with suspected DCD were assessed using the M-ABC-2 and the ZNA. Rank correlations between scores of different test components were calculated. The structure of the tests was explored using canonical-correlation analysis. The correlation between total scores of the two motor tests was reasonable (0.66; p<0.001). However, ZNA scores were generally lower than those of M-ABC-2, due to poor performance in the fine motor adaptive component and increased contralateral associated movements (CAM). The canonical-correlation analysis revealed that ZNA measures components like pure motor skills and CAM that are not represented in the M-ABC-2. Furthermore, there was also no equivalent for the aiming and catching items of the M-ABC-2 in ZNA. The two tests measure different motor characteristics in children with suspected DCD and, thus, can be used complementary for the diagnosis of the disorder.


Allelic loss of cyclophilin 40, an estrogen receptor-associated immunophilin, in breast carcinomas.
Cyclophilin 40 (CyP40) is an estrogen receptor-associated protein which appears to modify receptor function. The aim of this study was to determine the extent of allelic loss at the CyP40 locus in a panel of breast carcinomas using a newly characterized microsatellite marker located upstream of the CyP40 gene and then to correlate this with losses at chromosomal sites for cancer-associated genes. Allelic loss at CyP40 was determined from patients' matched tumor and normal breast tissue using Genescan 672 software analysis of fluorescently labeled, PAGE-separated PCR products incorporating the marker. For each patient, allelic loss at CyP40 was then assessed and compared with losses at markers for various cancer-associated genes. Allelic loss was detected in 30% of breast carcinomas from patients heterozygous for the CyP40 marker. All carcinomas demonstrating allelic loss were grade II or III invasive ductal carcinomas and generally showed multiple losses at other sites near known cancer-associated genes. The polymorphic marker which we characterized was useful in determining allelic loss at the CyP40 locus in breast cancer patients and when applied in these studies in conjunction with various cancer-associated gene markers, suggests that deletions in the region of the CyP40 gene might be a late event in breast tumor progression.


[Social representations of health and disease and the implications in nursing care: a structural analysis].
This study characterizes the social representations of the health-disease process of subjects resident in two districts of São Paulo, in order to identify the needs of health and the orientation of the nursing action. Free evocations recollections from 418 adults on the themes health and disease. Data analyse was developed thought a descriptive and structural analysis of the social representations, through the methodology of construction of the "picture of four houses", categorization and similitude analysis. The results show central senses of the representation, the possitiveness of the health, are anchored in a divine entity, and accompanied of notions that associate health to prevention of diseases, to biological needs, to activity and to which the attitude assumed to the disease. The representation structure of disease is similar, in inverse sense, to the one of the health: god is the center of the social representation, in its negative version, being expressed in the body through the pain, of the death and the inactivity and--in the spirit--through the sadness and of the depression. The discussion is that psychological character of health and of disease, that is expressed under the psychosocial needs committed, and the consequent need of rethinking the technological model of work in nursing.


Transfemoral tricuspid valve replacement and one-year outcomes: the TRISCEND study.
For patients with symptomatic, severe tricuspid regurgitation (TR), early results of transcatheter tricuspid valve (TV) intervention studies have shown significant improvements in functional status and quality of life associated with right-heart reverse remodelling. Longer-term follow-up is needed to confirm sustained improvements in these outcomes. The prospective, single-arm, multicentre TRISCEND study enrolled 176 patients to evaluate the safety and performance of transcatheter TV replacement in patients with ≥moderate, symptomatic TR despite medical therapy. Major adverse events, reduction in TR grade and haemodynamic outcomes by echocardiography, and clinical, functional, and quality-of-life parameters are reported to one year. Enrolled patients were 71.0% female, mean age 78.7 years, 88.0% ≥ severe TR, and 75.4% New York Heart Association classes III-IV. Tricuspid regurgitation was reduced to ≤mild in 97.6% (P < .001), with increases in stroke volume (10.5 ± 16.8 mL, P < .001) and cardiac output (0.6 ± 1.2 L/min, P < .001). New York Heart Association class I or II was achieved in 93.3% (P < .001), Kansas City Cardiomyopathy Questionnaire score increased by 25.7 points (P < .001), and six-minute walk distance increased by 56.2 m (P < .001). All-cause mortality was 9.1%, and 10.2% of patients were hospitalized for heart failure. In an elderly, highly comorbid population with ≥moderate TR, patients receiving transfemoral EVOQUE transcatheter TV replacement had sustained TR reduction, significant increases in stroke volume and cardiac output, and high survival and low hospitalization rates with improved clinical, functional, and quality-of-life outcomes to one year. Funded by Edwards Lifesciences, TRISCEND ClinicalTrials.gov number, NCT04221490.


Fecal Matrix Metalloprotease-9 and Lipocalin-2 as Biomarkers in Detecting Endoscopic Activity in Patients With Inflammatory Bowel Diseases.
Fecal biomarkers are emerging tools in the assessment of mucosal healing in inflammatory bowel diseases (IBD). We aimed to evaluate the accuracy of fecal matrix metalloprotease-9 (MMP-9) and fecal lipocalin-2 (LCN-2) compared with calprotectin in detecting endoscopic activity in IBD STUDY:: Overall, 86 IBD adults underwent colonoscopy consecutively and prospectively, with Crohn's disease Endoscopic Index of Severity (CDEIS) in Crohn's disease (CD) patients or Mayo endoscopic subscore calculation for ulcerative colitis (UC) patients, and stool collection. Fecal calprotectin was measured using quantitative immunochromatographic testing. Fecal MMP-9 and LCN-2 was quantified by enzyme-linked immunosorbent assay. MMP-9 and LCN-2 thresholds were determined using receiver operating curves. In 54 CD patients, fecal calprotectin, MMP-9 and LCN-2 correlated with CDEIS and were significantly increased in patients with endoscopic ulcerations. MMP-9 >350 ng/g detected endoscopic ulceration in CD with a sensitivity of 90.0% and a specificity of 63.6%, compared with fecal calprotectin >250 μg/g (sensitivity=90.5% and specificity=59.1%). Fecal LCN-2 demonstrated lower performances than the 2 other biomarkers (sensitivity=85.7% and specificity=45.5%).In 32 UC patients, fecal MMP-9, LCN-2, and calprotectin levels were significantly increased in patients with endoscopic activity. In UC patients, fecal MMP-9 >900 ng/g had the best efficacy to detect endoscopic activity (sensitivity=91.0% and specificity=80.0%, compared with fecal calprotectin >250 μg/g (sensitivity=86.4% and specificity=80.0%) and LCN-2 >6700 ng/g (sensitivity=82.0% and specificity=80.0%). Fecal MMP-9 is a reliable biomarker in detecting endoscopic activity in both UC and CD patients.


[The myoelectrical activity of the gastroduodenal area in ulcer formation due to serotonin].
The effects of prolonged subcutaneous administration of serotonin on the myoelectric activity of the gastroduodenal junction were investigated in conscious rabbits. Serotonin produced the duodenogastric discoordination by increasing duodenal activity and decreasing the activity of stomach and pylorus. This discoordination resulted in gastric ulceration. Atropine prevented both duodenogastric discoordination and ulceration. These results indicate that serotonin ulceration is related to duodenogastric discoordination.


Amino acid sequence of a new 2S albumin from Ricinus communis which is part of a 29-kDa precursor protein.
The isolation and sequence determination of a new 2S albumin storage protein from Ricinus communis seeds denoted 2S ASP-Ib are described. The fragment approach using selective enzymatic cleavage, Edman degradation, and mass spectrometry was used to demonstrate that the 11-kDa heterodimer protein linked by disulfide bridges has the following structure: short chain, GEREGSSSQQCRQEVQRKDLSSCERYLRQSSS; long chain, <QQQESQQLQQCCNQVKQVRDECQCEAIKYIAEDQIQQGQLHGEESERVAQRAGEIVSSCGVRCMR . The molecular weight of the intact protein, 11,140 +/- 2, determined by matrix-assisted laser desorption mass spectrometry was consistent with the assigned structure. The S- and L-chains are identical to residues 18-49 and 66-130 of the precursor protein predicted by S. D. Irwin, J. N. Keen, J. B. C. Findlay, and J. M. Lord [(1990) Mol. Gen. Genet. 222, 400-408], on the basis of the structure of a cDNA isolated using probes based on the sequence of another 2S albumin, described by F. S. Sharief and S. S. L. Li [(1982) J. Biol. Chem. 257, 14753-14759], which we denote 2S ASP-Ia. Three of the four termini could have been produced by posttranslational processing by endopeptidase(s) and carboxypeptidase(s) which utilized basic residues as the cleavage sites. Mass spectrometric evidence suggested that the protein presented microheterogeneity at its termini, i.e., truncated forms presumably due to processing heterogeneity. The present characterization of the 2S ASP-Ib protein, the second 2S albumin from Ricinus communis seeds, demonstrates that the 237-residue precursor protein codes for two different heterodimer proteins containing 97 and 99 residues each. This system should be useful for studying the posttranslational processing of plant storage proteins.


Protection against feline leukemia virus infection by use of an inactivated virus vaccine.
The protective immunity induced by 3 experimental FeLV vaccines were evaluated: Prototype inactivated FeLV vaccine developed from a molecularly cloned FeLV isolate (FeLV-FAIDS-61E-A); a mixture of immunodominant synthetic peptides corresponding to regions of the FeLV-Gardner-Arnstein-B (FeLV-GA-B) envelope proteins; and an adjuvant-disrupted but non-activated virus prepared from a non-cloned FeLV field isolate comprised of subgroup A and B viruses (FeLV-05821-AB). Included as controls were parallel groups of cats inoculated with adjuvants alone or with an established commercial FeLV vaccine. After each inoculation and after virulent virus challenge exposure, sera from all cats were assayed for ELISA-reactive antibody against purified FeLV, FeLV neutralizing (VN) antibody, and FeLV antigenemia/viremia--viral p27 antigen in serum and within circulating leukocytes. Immunity was challenged by oral/nasal exposure of vaccinated and control cats with FeLV-FAIDS-61E-A or FeLV-05821-AB, an infective, noncloned, tissue-origin, FeLV field isolate containing subgroup-A and -B viruses. Vaccine-induced immunity was assessed by comparing the postchallenge-exposure incidence of persistent viremia and the pre- and postchallenge exposure titers of VN and ELISA antibody in cats of the control and vaccine groups. The percentage of cats, that resisted development of persistent viremia after FeLV challenge exposure and the preventable fraction (PF) for the vaccine groups (which adjusts for the severity of the challenge and the degree of innate resistance in the controls) were as follows: adjuvant controls, 26%; FeLV-FAIDS-61E-A inactivated virus vaccine, 95% (PF = 93.2%); FeLV-GA-B peptide vaccine, 5% (-28.4%); FeLV-05821-AB noninactivated vaccine, 67% (55.4%); and commercial FeLV vaccine, 35% (12.2%). The prechallenge exposure mean VN antibody titer for each group was: less than 1:8 in the adjuvant controls; 1:43 in the FeLV-FAIDS-61E-A-vaccinated cats; less than 1:8 in the peptide-vaccinated cats; 1:38 in the noninactivated virus-vaccinated cats group; and 1:12 in the cats vaccinated with the commercial vaccine. Thus, induction of VN antibody in the vaccinated cats, although modest, appeared to be correlated with induction of protective immunity as defined by resistance to FeLV challenge exposure. Results of these studies indicate that inoculation of cats with an experimental inactivated virus vaccine prepared from a molecularly cloned FeLV isolate was most effective in stimulating protective immunity against heterologous and homologous FeLV challenge exposure.


Evaluation of meteorological and epidemiological characteristics of fatal pulmonary embolism.
The objective of the present study was to identify risk factors among epidemiological factors and meteorological conditions in connection with fatal pulmonary embolism. Information was collected from forensic autopsy records in sudden unexpected death cases where pulmonary embolism was the exact cause of death between 2001 and 2010 in Budapest. Meteorological parameters were detected during the investigated period. Gender, age, manner of death, cause of death, place of death, post-mortem pathomorphological changes and daily meteorological conditions (i.e. daily mean temperature and atmospheric pressure) were examined. We detected that the number of registered pulmonary embolism (No 467, 211 male) follows power law in time regardless of the manner of death. We first described that the number of registered fatal pulmonary embolism up to the nth day can be expressed as Y(n) = α ⋅ n (β) where Y denotes the number of fatal pulmonary embolisms up to the nth day and α > 0 and β > 1 are model parameters. We found that there is a definite link between the cold temperature and the increasing incidence of fatal pulmonary embolism. Cold temperature and the change of air pressure appear to be predisposing factors for fatal pulmonary embolism. Meteorological parameters should have provided additional information about the predisposing factors of thromboembolism.


The kinetics of ethylmorphine N-demethylation in isolated hepatocytes: the effect of drug transport and oxygen concentration.
In previous studies we have observed that the Km for ethylmorphine N-demethylase was only 50 to 100 microM in the isolated hepatocyte as opposed to 250 to 340 microM in isolated microsomes. These data suggested that either the drug is actively transported into the cell or that the enzymes in the cell had a reduced turnover number. Our current study indicates that ethylmorphine is transported into the hepatocyte with a Km of 250 microM and Tm of 20 nmol/min 10(6) cells. On the basis of temperature and inhibitor studies, this would appear to be active transport. When the turnover number of the ethylmorphine N-demethylase was decreased by reducing the oxygen from 20% of the gas phase to 5%, the activity was partially uncompetitively inhibited with the Km decreasing from 182 to 105 microM. The Vm decreased from 1.85 nmol/min 10(6) to 0.71 nmol/min 10(6) cells. These data suggest that the low Km we initially observed was probably due to active transport of the substrate into the hepatocyte. Further, our observations indicate that the effects of oxygen pressure on the kinetic parameters should be considered in predicting in vivo kinetics from in vitro studies.


Acute ethanol effects on local cerebral glucose utilization in select central nervous system regions of adolescent alcohol-preferring (P) and alcohol-nonpreferring (NP) rats.
Alcohol abuse among adolescents is a major health and developmental problem. The 2-[(14)C]deoxyglucose (2-DG) technique allows for the in vivo quantification of local cerebral glucose utilization (LCGU) as a measure of functional neuronal activity. Local cerebral glucose utilization rates were examined after acute ethanol administration within selected brain regions of adolescent alcohol-preferring (P) and -nonpreferring (NP) rats. Postnatal day 45 male P and NP rats were injected with saline or 1.0 g/kg ethanol, i.p., 10 minutes prior to an intravenous bolus of [(14)C]-2-deoxyglucose (125 microCi/kg). Image densities were determined using quantitative autoradiography and LCGU values calculated. Acute ethanol injection significantly decreased LCGU rates in select brain regions including the olfactory tubercles, the frontal cortex (Fr), and subregions of the posterior hippocampus (pCA1 and pCA3). Acute ethanol had no significant effects on LCGU rates in any region of the adolescent NP rats. Significant basal LCGU rate differences were apparent between the rat lines in a nearly global fashion with adolescent P rats having much higher basal LCGU rates compared with adolescent NP rats. These findings suggest that the adolescent P and NP rats are less sensitive to the effects of acute ethanol than their adult counterparts. The adolescent P rat is relatively more sensitive to the initial effects of acute ethanol in select brain regions as compared with the adolescent NP rat. Additionally, the innate hyper-excited state of the adolescent P central nervous system is a likely factor in the development of their high alcohol drinking behaviors.


Point-of-care testing with high-sensitivity cardiac troponin assays: the challenges and opportunities.
Methods to improve the safety, accuracy and efficiency of assessment of patients with suspected acute coronary symptoms have occupied decades of study and have supported significant changes in clinical practice. Much of the progress is reliant on results of laboratory-based high-sensitivity cardiac troponin assays that can detect low concentrations with high precision. Until recently, point-of-care (POC) platforms were unable to perform with similar analytical precision as laboratory-based assays, and recommendations for their use in accelerated assessment strategies for patients with suspected acute coronary syndrome has been limited. As POC assays can provide troponin results within 20 min, and can be used proximate to patient care, improvements in the efficiency of assessment of patients with suspected acute coronary syndrome is possible, particularly with new high-sensitivity assays.


Nitric oxide prevents H2O2-induced apoptosis in SK-N-MC human neuroblastoma cells.
Nitric oxide (NO) is a cellular signaling molecule in many physiological and pathological processes including neuroprotector. Here we examined the antiapoptotic effect of NO in SK-N-MC cells. H2O2 treatment (10-200 μM) induced cell death in a dose-dependent manner and pretreatment of cells with 100 μM S-nitroso-N-acetylpenicillamine (SNAP), an NO donor, attenuated the occurrence of H2O2-induced cell death. DAPI staining showed H2O2-induced nuclear fragmentation and NO treatment suppressed it. NO inhibited the proteolytic activation of caspase-3 and mitochondrial cytochrome c release. Treatment of soluble guanylyl cyclase inhibitor ODQ decreased the protective effect of SNAP on H2O2-treated cells and increased caspase 3-like enzyme activity and activation, cytochrome c release, PARP cleavage, and DNA fragmentation, indicating that cGMP is a key mediator in NO-mediated antiapoptosis. The cGMP analog 8-Br-cGMP blocked H2O2-induced apoptotic cell death; reduction of caspase-3 enzyme, cytochrome c release, and caspase-8 and -9. These preventive effects of SNAP and 8-Br-cGMP were suppressed by PKG inhibitor KT5823. Levels of PKGI, PKGII, and p-VASP proteins were increased by SNAP and 8-Br-cGMP and suppressed by KT5823 treatment. These results indicate that PKG is a downstream signal mediator in the suppression of apoptosis by NO and cGMP. Akt activation was inhibited the PI3K inhibitors LY294002 and Wortmannin, resulting in the inhibition of cell viability and increase of cytochrome c release. SNAP induced phosphorylation of Akt and Bad and then increased the interactions between 14-3-3β and p-Bad. These data suggest that the NO suppresses H2O2-induced SK-N-MC cell apoptosis by suppressing apoptosis signal mediating the interaction between 14-3-3β and Bad phosphorylation via PKG/PI3K/Akt.


Adrenomedullin is a potent stimulator of osteoblastic activity in vitro and in vivo.
Adrenomedullin is a 52-amino acid vasodilator peptide produced in many tissues, including bone. It has 20% sequence identity with amylin, a regulator of osteoblast growth, and circulates in picomolar concentrations. The present study assesses whether adrenomedullin also acts on osteoblasts. At concentrations of 10(-12) M and greater, adrenomedullin produced a dose-dependent increase in cell number and [3H]thymidine incorporation in cultures of fetal rat osteoblasts. This effect was also seen with adrenomedullin-(15-52), -(22-52), and -(27-52), but adrenomedullin-(40-52) was inactive. These effects were lost in the presence of amylin blockers, suggesting they were mediated by the amylin receptor. Adrenomedullin also increased [3H]thymidine incorporation into cultured neonatal mouse calvaria but, unlike amylin, did not reduce bone resorption in this model. Adrenomedullin stimulated phenylalanine incorporation into both isolated osteoblasts and calvaria. When injected daily for 5 days over the calvariae of adult mice, it increased indexes of bone formation two- to threefold (P < 0.0001) and increased mineralized bone area by 14% (P = 0.004). It is concluded that adrenomedullin regulates osteoblast function and that it increases bone mass in vivo. The potential of this family of peptides in the therapy of osteoporosis should be further evaluated.


Psychological factors associated with response to treatment in rheumatoid arthritis.
This paper presents a comprehensive review of research relating psychological domains with response to therapy in patients with rheumatoid arthritis. A holistic approach to the disease was adopted by incorporating not only disease activity but also dimensions of the impact of disease on patients' lives. Psychological distress, including depression and anxiety, is common among patients with rheumatoid arthritis and has a significant negative impact on response to therapy and on patients' abilities to cope with chronic illness. Evidence regarding the influence of positive psychological dimensions such as acceptance, optimism, and adaptive coping strategies is scarce. The mechanisms involved in these interactions are incompletely understood, although changes in neuro-endocrine-immune pathways, which are common to depression and rheumatoid arthritis, seem to play a central role. Indirect psychological influences on therapeutic efficacy and long-term effectiveness include a myriad of factors such as adherence, placebo effects, cognition, coping strategies, and family and social support. Data suggest that recognition and appropriate management of psychological distress may improve response to treatment and significantly reduce disease burden.


Caretaker or undertaker? The role of the proteasome in aging.
Despite intensive studies, the molecular basis of the decline of protein degradation with age still remains unresolved. It is suspected that the proteasome is one of the key factors controlling the age-dependent turnover of intracellular proteins. This hypothesis is based on the observation that the proteasome is a part of the ubiquitin-proteasome pathway, which together with the lysosomal pathway constitute the major mechanisms of protein degradation. While there are alterations in proteasome structure and function with age, the observed changes do not provide a clear mechanism for explaining the decline of protein degradation. In addition, there are no consistent changes in the ubiquitination system to account for this decline. On the other hand, because of the essential role played by the proteasome in the maintenance of cellular homeostasis, the observation of age-related changes in structure and function will ultimately be demonstrated to contribute to the aging process. The fact that food restriction, the only currently available experimental paradigm that can alter the aging process, modulates the age-related changes in proteasome structure and function provides presumptive evidence that the proteasome is involved in the aging process.


Should scatter be corrected in both transmission and emission data for accurate quantitation in cardiac SPET?
Ideally, reliable quantitation in single-photon emission tomography (SPET) requires both emission and transmission data to be scatter free. Although scatter in emission data has been extensively studied, it is not well known how scatter in transmission data affects relative and absolute quantitation in reconstructed images. We studied SPET quantitative accuracy for different amounts of scatter in emission and transmission data using a Utah phantom and a cardiac Data Spectrum phantom including different attenuating media. Acquisitions over 180 degrees were considered and three projection sets were derived: 20% images and Jaszczak and triple-energy-window scatter-corrected projections. Transmission data were acquired using gadolinium-153 line sources in a 90-110 keV window using a narrow or wide scanning window. The transmission scans were performed either simultaneously with the emission acquisition or 24 h later. Transmission maps were reconstructed using filtered backprojection and mu values were linearly scaled from 100 to 140 keV. Attenuation-corrected images were reconstructed using a conjugate gradient minimal residual algorithm. The mu value underestimation varied between 4% with a narrow transmission window in soft tissue and 22% with a wide window in a material simulating bone. Scatter in the emission and transmission data had little effect on the uniformity of activity distribution in the left ventricle wall and in a uniformly hot compartment of the Utah phantom. Correcting the transmission data for scatter had no impact on contrast between a hot and a cold region or on signal-to-noise ratio (SNR) in regions with uniform activity distribution, while correcting the emission data for scatter improved contrast and reduced SNR. For absolute quantitation, the most accurate results (bias <4% in both phantoms) were obtained when reducing scatter in both emission and transmission data. In conclusion, trying to obtain the same amount of scatter in emission and transmission data, in addition to being impractical because of the difficulty in knowing the precise scatter components, did not yield such accurate absolute activity quantitation as when emission and transmission scatter were reduced.


Multirelaxation-time interaction-potential-based lattice Boltzmann model for two-phase flow.
The application of the lattice Boltzmann method in two-phase flows is often restricted by the numerical instability at low viscosities. In this work, a multirelaxation-time (MRT) lattice Boltzmann model (LBM) is developed using the interaction potential approach. With the MRT collision term and a general force term, the new MRT model is able to significantly enhance the numerical stability at low viscosities, without appreciable increase in computation time or memory use. Advanced force formulation using the multirange potential can also be readily incorporated into the current MRT scheme. Numerical tests are first performed in two dimensions under equilibrium conditions. The MRT model is able to reduce the lowest stable viscosity by an order of magnitude compared to the single relaxation time LBM. In addition, the spurious velocity at the gas-liquid interface can also be significantly decreased by tuning the adjustable relaxation parameters. Then two sets of three-dimensional simulations are conducted to investigate the buoyant rise of a gas bubble in a low-viscosity liquid. In particular, millimeter air bubble in water, which is difficult for traditional two-phase LBM due to both low viscosity and high-surface tension, is successfully simulated using the MRT technique developed in this study. The simulated bubble shape and velocity are compared with the experimental results and empirical correlations in the literature, and the satisfactory agreement proves the validity of the MRT-LBM for two-phase flows.


Vibrational mode multiplexing of ultracold atoms.
Sending multiple messages on qubits encoded in different vibrational modes of cold atoms or ions along a transmission waveguide requires us to merge first and then separate the modes at input and output ends. Similarly, different qubits can be stored in the modes of a trap and be separated later. We design the fast splitting of a harmonic trap into an asymmetric double well so that the initial ground vibrational state becomes the ground state of one of two final wells, and the initial first excited state becomes the ground state of the other well. This might be done adiabatically by slowly deforming the trap. We speed up the process by inverse engineering a double-function trap using dynamical invariants. The separation (demultiplexing) followed by an inversion of the asymmetric bias and then by the reverse process (multiplexing) provides a population inversion protocol based solely on trap reshaping.


Use of Noncontrast Computed Tomography and Computed Tomographic Perfusion in Predicting Intracerebral Hemorrhage After Intravenous Alteplase Therapy.
Intracerebral hemorrhage is a feared complication of intravenous alteplase therapy in patients with acute ischemic stroke. We explore the use of multimodal computed tomography in predicting this complication. All patients were administered intravenous alteplase with/without intra-arterial therapy. An age- and sex-matched case-control design with classic and conditional logistic regression techniques was chosen for analyses. Outcome was parenchymal hemorrhage on 24- to 48-hour imaging. Exposure variables were imaging (noncontrast computed tomography hypoattenuation degree, relative volume of very low cerebral blood volume, relative volume of cerebral blood flow ≤7 mL/min·per 100 g, relative volume of Tmax ≥16 s with all volumes standardized to z axis coverage, mean permeability surface area product values within Tmax ≥8 s volume, and mean permeability surface area product values within ipsilesional hemisphere) and clinical variables (NIHSS [National Institutes of Health Stroke Scale], onset to imaging time, baseline systolic blood pressure, blood glucose, serum creatinine, treatment type, and reperfusion status). One-hundred eighteen subjects (22 patients with parenchymal hemorrhage versus 96 without, median baseline NIHSS score of 15) were included in the final analysis. In multivariable regression, noncontrast computed tomography hypoattenuation grade (P<0.006) and computerized tomography perfusion white matter relative volume of very low cerebral blood volume (P=0.04) were the only significant variables associated with parenchymal hemorrhage on follow-up imaging (area under the curve, 0.73; 95% confidence interval, 0.63-0.83). Interrater reliability for noncontrast computed tomography hypoattenuation grade was moderate (κ=0.6). Baseline hypoattenuation on noncontrast computed tomography and very low cerebral blood volume on computerized tomography perfusion are associated with development of parenchymal hemorrhage in patients with acute ischemic stroke receiving intravenous alteplase.


Differential Gene Expression of Malaria Parasite in Response to Red Blood Cell-Specific Glycolytic Intermediate 2,3-Diphosphoglycerate (2,3-DPG).
Innovative strategies to control malaria are urgently needed. Exploring the interplay between Plasmodium sp. parasites and host red blood cells (RBCs) offers opportunities for novel antimalarial interventions. Pyruvate kinase deficiency (PKD), characterized by heightened 2,3-diphosphoglycerate (2,3-DPG) concentration, has been associated with protection against malaria. Elevated levels of 2,3-DPG, a specific mammalian metabolite, may hinder glycolysis, prompting us to hypothesize its potential contribution to PKD-mediated protection. We investigated the impact of the extracellular supplementation of 2,3-DPG on the Plasmodium falciparum intraerythrocytic developmental cycle in vitro. The results showed an inhibition of parasite growth, resulting from significantly fewer progeny from 2,3-DPG-treated parasites. We analyzed differential gene expression and the transcriptomic profile of P. falciparum trophozoites, from in vitro cultures subjected or not subjected to the action of 2,3-DPG, using Nanopore Sequencing Technology. The presence of 2,3-DPG in the culture medium was associated with the significant differential expression of 71 genes, mostly associated with the GO terms nucleic acid binding, transcription or monoatomic anion channel. Further, several genes related to cell cycle control were downregulated in treated parasites. These findings suggest that the presence of this RBC-specific glycolytic metabolite impacts the expression of genes transcribed during the parasite trophozoite stage and the number of merozoites released from individual schizonts, which supports the potential role of 2,3-DPG in the mechanism of protection against malaria by PKD.


Lower- extremity biomechanics and maintenance of vertical-jump height during prolonged intermittent exercise.
Potential biomechanical compensations allowing for maintenance of maximal explosive performance during prolonged intermittent exercise, with respect to the corresponding rise in injury rates during the later stages of exercise or competition, are relatively unknown. To identify lower-extremity countermovement-jump (CMJ) biomechanical factors using a principal-components approach and then examine how these factors changed during a 90-min intermittent-exercise protocol (IEP) while maintaining maximal jump height. Mixed-model design. Laboratory. Fifty-nine intermittent-sport athletes (30 male, 29 female) participated in experimental and control conditions. Before and after a dynamic warm-up and every 15 min during the 1st and 2nd halves of an individually prescribed 90-min IEP, participants were assessed on rating of perceived exertion, sprint/cut speed, and 3-dimensional CMJ biomechanics (experimental). On a separate day, the same measures were obtained every 15 min during 90 min of quiet rest (control). Univariate piecewise growth models analyzed progressive changes in CMJ performance and biomechanical factors extracted from a principal-components analysis of the individual biomechanical dependent variables. While CMJ height was maintained during the 1st and 2nd halves, the body descended less and knee kinetic and energetic magnitudes decreased as the IEP progressed. The results indicate that vertical-jump performance is maintained along with progressive biomechanical changes commonly associated with decreased performance. A better understanding of lower-extremity biomechanics during explosive actions in response to IEP allows us to further develop and individualize performance training programs.


Effects of intraoperative reading on vigilance and workload during anesthesia care in an academic medical center.
During routine cases, anesthesia providers may divert their attention away from direct patient care to read clinical (e.g., medical records) and/or nonclinical materials. The authors sought to ascertain the incidence of intraoperative reading and measure its effects on clinicians' workload and vigilance. In 172 selected general anesthetic cases in an academic medical center, a trained observer categorized the anesthesia provider's activities into 37 possible tasks. Vigilance was assessed by the response time to a randomly illuminated alarm light. Observer- and subject-reported workload were scored at random intervals. Data from Reading and Non-Reading Periods of the same cases were compared to each other and to matched cases that contained no observed reading. The cases were matched before data analysis on the basis of case complexity and anesthesia type. Reading was observed in 35% of cases. In these 60 cases, providers read during 25 +/- 3% of maintenance but not during induction or emergence. While Non-Reading Cases (n = 112) and Non-Reading Periods of Reading Cases did not differ in workload, vigilance, or task distribution, they both had significantly higher workload than Reading Periods. Vigilance was not different among the three groups. When reading, clinicians spent less time performing manual tasks, conversing with others, and recordkeeping. Anesthesia providers, even when being observed, read during a significant percentage of the maintenance period in many cases. However, reading occurred when workload was low and did not appear to affect a measure of vigilance.


Sex-related differences in parental rearing patterns in young adults with bipolar disorder.
The aim of this study was to examine the parenting characteristics of young patients with bipolar disorder (BD) and explore the sex differences. The parental rearing pattern of young patients with BD was measured and compared with the healthy control of young adults. The EMBU scale was used to assess parental rearing patterns. Patients with BD reported significantly higher scores in the punishment and severity index, as well as of the rejection and denial index, but lower scores in the warmth & affectionate index in the paternal rearing pattern, compared with healthy controls. In addition, patients scored higher on the punishment and severity index and rejection and patterns index in maternal rearing patterns. More importantly, we found significant sex differences in maternal rearing patterns (pBonferroni < 0.05). Specifically, in the maternal rearing patterns, male patients had higher scores on the favoring index than male controls, whereas female patients had lower scores on the warmth & affectionate index than female controls. This study shows significant differences in parental rearing patterns between patients and control subjects. Male patients were overprotective by their mothers and female patients were overlooked by their mothers during upbringing.


Enhanced podocyte differentiation and changing drug toxicity sensitivity through pressure-controlled mechanical filtration stress on a glomerulus-on-a-chip.
Podocytes, localized in the glomerulus, are a prognostic factor of proteinuria in kidney disease and are exposed to distinct physiological stimuli from basal to apical filtration flow. Research studies on drug discovery and disease modeling for glomerulopathy have developed a glomerulus-on-a-chip and studied podocyte mechanobiology to realize alternative methods to animal experiments. However, the effect of filtration stimulus on podocytes has remained unclear. Herein, we report the successful development of a user-friendly filtration culture device and system that can precisely control the filtration flow using air pressure control by incorporating a commercially available culture insert. It allows mouse podocytes to be cultured under filtration conditions for three days with a guarantee of maintaining the integrity of the podocyte layer. Using our system, this study demonstrated that podocyte damage caused by hyperfiltration resulting from glomerular hypertension, a common pathophysiology of many glomerulopathies, was successfully recapitulated and that filtration stimulus promotes the maturation of podocytes in terms of their morphology and gene expression. Furthermore, we demonstrated that filtration stimulus induced different drug responsiveness in podocytes than those seen under static conditions, and that the difference in drug responsiveness was dependent on the pharmacological mechanism. Overall, this study has revealed differentiating and pharmacodynamic properties of filtration stimulus and brings new insights into the research field of podocyte mechanobiology towards the realization of glomerulus-on-a-chip.


Volume-induced natriuresis in healthy women: renal metabolism of prostacyclin and thromboxane, and physiological role of prostanoids.
In healthy women submitted to a short-term expansion in extracellular fluid volume we have evaluated the urinary excretory profile of the stable metabolites of prostaglandin(PG) I2 and thromboxane(TX) A2, 6-keto-PGF1 alpha(6KPGF) and TXB2 respectively, and assessed the physiological role played by the prostanoids in this experimental condition. Salt retention (SR group, n=9) was induced by repeated i.v. infusion of saline solution (0.9% NaCl). At the end of the treatment the body weight had increased by 0.7+/-0.2 kg (mean+/-SEM) (P<0.05). Renal functional exploration [clearance (cl.) method] was performed during hypotonic polyuria (induced by oral water load) and subsequent moderate antidiuresis (induced by low-dose infusion of an antidiuretic hormone analogue). Urinary 6KPGF and TXB2 concentrations were estimated by RIA method during polyuria (P cl. period), early and late antidiuresis (A1 and A2 cl. periods). Paired functional explorations were performed in absence (control study) and presence of indomethacin. Basal values of plasma sodium and potassium concentrations, plasma renin activity (PRA) and urinary aldosterone excretion were determined just before the control study. The results in salt retention were compared to those previously obtained in healthy women submitted to a moderate salt depletion (SD2 group, n=6), in absence and presence of the drug. Women in salt retention received 100 mg i.m. of the drug, whereas salt-depleted women received only a halved dose as in previous studies in salt depletion the full dose produced prolonged anuria. (I) Salt retention vs salt depletion. The basal values of PRA and urinary aldosterone excretion were significantly lower. During polyuria, urinary excretion of 6KPGF, 6KPGF/TXB2 ratio, urinary flow rate, creatinine cl. and absolute and fractional excretions of sodium and chloride were significantly higher. In salt retention during polyuria, significant positive correlations were found between 6KPGF excretion and functional excretory parameters. (II) Indomethacin in salt retention. The following effects were significant: (a) a reduction in prostanoid excretions in P and A1 cl. periods only; (b) during polyuria, an increase in arterial pressure, a reduction in urinary flow rate and creatinine cl. (saluresis showed not significant reduction). During polyuria significant positive correlations occurred between the absolute effects of indomethacin on 6KPGF excretion and those on functional excretory parameters. (III) Comparative effects of indomethacin in salt retention and salt depletion. Despite the double dosage of the drug, the significant reductions in urinary metabolite excretions were not significantly different during P cl. period and significantly lower in A1 cl. period compared to the corresponding significant reductions in salt depletion. During polyuria, the significant increase in arterial pressure was significantly different from the not significant effect in salt depletion; the not significant effect on saluresis was significantly different from the significant reduction in salt depletion. The results suggest the following conclusions: (1) The present model showed the functional pattern of the volume-natriuresis; (2) In salt retention, in contrast with salt depletion, indomethacin induced an increase in arterial pressure consistent with the inhibition of a PG-dependent vasodilator mechanism active at the systemic level; (3) In salt retention, in contrast with salt depletion, indomethacin failed to induce a significant reduction in saluresis. This failure can be attributed to the drug's blunted effectiveness in inhibiting the renal synthesis of saluretic PGs, and probably to the interference of the concurrent increase in arterial pressure in the renal treatment of sodium and chloride.


Multisensory integration in complete unawareness: evidence from audiovisual congruency priming.
Multisensory integration is thought to require conscious perception. Although previous studies have shown that an invisible stimulus could be integrated with an audible one, none have demonstrated integration of two subliminal stimuli of different modalities. Here, pairs of identical or different audiovisual target letters (the sound /b/ with the written letter "b" or "m," respectively) were preceded by pairs of masked identical or different audiovisual prime digits (the sound /6/ with the written digit "6" or "8," respectively). In three experiments, awareness of the audiovisual digit primes was manipulated, such that participants were either unaware of the visual digit, the auditory digit, or both. Priming of the semantic relations between the auditory and visual digits was found in all experiments. Moreover, a further experiment showed that unconscious multisensory integration was not obtained when participants did not undergo prior conscious training of the task. This suggests that following conscious learning, unconscious processing suffices for multisensory integration.


Seroprevalence of antibodies against serogroup C meningococci in England in the postvaccination era.
The United Kingdom introduced meningococcal serogroup C conjugate (MCC) vaccines in 1999, resulting in substantial declines in serogroup C disease and carriage. Here, we measured the age-specific prevalence of serum bactericidal antibodies (SBA) to Neisseria meningitidis serogroup C and immunoglobulin G (IgG) concentrations to serogroups A, C, W-135, and Y in 2,673 serum samples collected in England between 2000 and 2004. We compared the seroprevalence of SBA titers of > or =8 in the postvaccination era with results from an earlier prevaccination study conducted using the same methods. We found that the percentages of individuals with protective SBA titers were higher in 2000 to 2004 in all of the age groups targeted for MCC vaccination. In the postvaccine era, the prevalence of protective titers was high (75%) in children who had recently been offered routine immunization, but this fell to 36% more than 18 months after scheduled immunization. In the cohorts targeted in the catch-up campaign, the percentage achieving SBA titers of > or =8 was higher in children offered the vaccine at ages 5 to 17 years than in children offered the vaccine at ages 1 to 4 years. The geometric mean concentration (GMC) IgG for serogroup C followed a similar pattern, corresponding to the age at and time since scheduled MCC vaccination. Serogroup-specific IgG GMCs for W-135 and Y were low and showed little variation by age. Serogroup A IgG GMCs were higher, possibly reflecting exposure to cross-reacting antigens. Although the incidence of serogroup C disease remains low due to persisting herd effects, population antibody levels to serogroup C meningococci should be monitored so that potentially susceptible age groups can be identified should herd immunity wane.


Xenogenic cancer vaccines.
In the current review the latest literature data on design of cancer vaccines on the basis of xenogenic tumor-associated antigens and proteins that play an important role in tumor progression have been summarized. The main benefits and disadvantages of xenogenic cancer vaccines, the results of experimental studies and clinical trials, and perspectives of use are analyzed.


Determination of patterning in the pollen wall of Lilium henryi.
Centrifugation of living pollen mother cells of Lilium henryi has permitted the modifications of complex walls developed on the four daughter microspores. This experimental approach has not only provided an insight into the mechanism by which patterning is established, but has also identified the stages in cell development when wall determinants are present. These determinants can be first detected late in the meiotic prophase, but are not necessarily active at the cell surface at this early stage. The fact that treatment during cytokinesis may affect subsequent wall formation, combined with the obvious interference of the colpus with the spore patterning, indicates pattern determination to be a lengthy process, commencing during late prophase and reaching completion in the young tetrad. The study of walls formed on enucleate cell fragments suggests that neither the spindle nor the cytoskeleton is intimately involved in formation of the basic reticulate patterning. It is proposed that material is inserted into the mosaic of the plasma membrane and is reorganized biophysically to form contiguous 'plates', which then modify the properties of the membrane in such a way that the cytoplasmic protrusions that constitute the first stages of wall formation only develop in the interstices between them. Changes in the plasma membrane have been detected over this period and it seems that a population of coated vesicles is closely involved in these events. Examination of the cytoskeleton of the young spore suggests that microtubules play a critical rôle in the positioning of the colpus, and reveals ways by which only small changes in this skeleton could give rise to multicolporate grains.


An in vitro perfusion system to examine the responses of endothelial cells to simulated flow and inflammatory stimulation.
Atherosclerosis is a disease process which develops at the arterial branches and curvatures of medium to large arteries. Local haemodynamic flow patterns in these vessels play an essential role in the formation of atherosclerotic lesions. To simulate pro-atherogenic blood flow patterns, we have developed a perfusion system with the ability to simulate in vivo patterns of blood flow in vitro. In this system, human umbilical vein endothelial cells were seeded in y-shaped microslides, in which they were exposed to a variety of flow patterns. Besides being able to reproduce the disturbed flow involved in the development of pro-atherogenic events within the arterial wall, the system also permitted the assessment of the pre-conditioning/priming effect of oscillatory flow on endothelial cells. The system was further capable of integrating multi-endpoint assays relevant to cardiovascular disease. We show that oscillatory flow primed endothelial cells, making them more sensitive to subsequent treatments. The treatment of oscillatory flow primed cells with TNFalpha resulted in the detection of enhanced levels of pro-inflammatory and chemoattractant factors such as IL-8 and MCP-1. These measurements were facilitated by the small volumes of medium circulating within the perfusion system. Oscillatory flow also altered the characteristics of monocyte adhesion to the endothelial layer. In summary, this system allows the monitoring of multiple endpoints and biomarkers, and provides an alternative to the use of in vivo and ex vivo models of cardiovascular disease.


Use of the deoxyuridine suppression test to evaluate vitamin B12 and folate status.
The deoxyuridine suppression test (DUST), performed on bone marrow cells, or peripheral blood lymphocytes, provides a rapid, dynamic assessment of vitamin B12 and folate status. The principles of this test are described and the use of the DUST in haematological practice at Wellington Hospital is reviewed. The advantages of the test are the speed of obtaining results, and an accurate assessment of the patient's condition when other haematological tests may be misleading.


Human ocular biometry.
The aim of this study was to examine growth of the human eye globe and cornea from early in gestation to late in adult life. Globe antero-posterior length, horizontal and vertical diameters, corneal horizontal and vertical (white to white) diameters and posterior pole to limbus distances were measured using digital calipers (±0.01 mm) in 541 postmortem eyes. Additional pre- and postnatal data for some of the dimensions were obtained from the literature. All dimensions examined increase rapidly during prenatal development but postnatal growth differs. Growth of globe antero-posterior length, vertical and horizontal diameters as well as corneal vertical and horizontal diameters stops within 1 year after birth. Logistic analysis is consistent with an asymptotic prenatal growth mode and no further growth after its completion around 1 year after birth. Horizontal and vertical globe diameters are the same at all ages but the corneal horizontal diameter is always larger than the vertical diameter. No differences could be detected between males and females in any of the ocular dimensions. Globe and corneal growth take place primarily during the prenatal growth mode and dimensions reach their maxima, shortly after birth. It is suggested that cessation of a growth stimulating signal at birth marks the end of the prenatal growth mode and that the small increases over the next year are due to cells already stimulated. Male and female eyes of the same age have the same globe and cornea dimensions.


The promoter of the Arabidopsis thaliana Cel1 endo-1,4-beta glucanase gene is differentially expressed in plant feeding cells induced by root-knot and cyst nematodes.
SUMMARY Transgenic tobacco and Arabidopsis thaliana carrying the Arabidopsis endo-1,4-beta-glucanase (EC 188.8.131.52) Cel1 promoter fused to the beta-glucuronidase (GUS) reporter gene were infected with the root-knot nematode, Meloidogyne incognita, and either the tobacco cyst nematode, Globodera tabacum (tobacco), or beet cyst nematode, Heterodera schachtii (Arabidopsis). Cel1-driven GUS expression was detected in cell elongation zones of noninfected plants and within feeding sites (giant-cells) induced in roots of both plant hosts by M. incognita. The first detectable signs of Cel1 expression within developing giant-cells occurred at the onset of giant-cell formation and continued throughout the M. incognita life cycle. UidA (Gus) transcripts were detectable within giant-cells induced in tobacco roots at 11-13 days postinoculation with M. incognita as determined by in situ mRNA hybridization. By contrast, expression of the Cel1 promoter was not detected within developing syncytia induced in tobacco or Arabidopsis roots by G. tabacum and H. schachtii, respectively, at any time point. The results demonstrate specific regulation of cell wall-degrading enzymes that may be required for cell wall modifications during feeding cell formation by sedentary endoparasitic nematodes. Differential expression of Cel1 by cyst and root-knot nematodes further supports underlying mechanistic differences in giant-cell and syncytium formation.


Prediction breeding value and genetic parameter in Iranian Holstein bulls for milk production traits.
Data set included records of 43303 cows for first lactation. Records were collected from 2000 to 2006 in Animal Breeding Center of Iran Studied traits were yield of milk, fat, protein and percentage of fat and protein. Total number of animal was 197561 individual in pedigree. Genetic and phenotypic parameters were estimated with REML method under single trait Animal Model. Breeding values were predicted with BLUP procedure. The model for the analyses included the factors herd-year-season as fixed factor (1694 levels), animal as random effect and age at calving as co variable with minimum and maximum of 23 and 36 month. Respectively the estimated heritabilities were 0.35 (+/-0.02), 0.33 (+/-0.02), 0.31 (+/-0.017), 0.28 (+/-0.02), 0.27 (+/-0.016) for milk, fat, protein yield, percent of fat and protein. Mean of breeding values of sires were 180.2 (+/-28.2), 3.7 (+/-1.26), 2.3 (+/-1.06), -0.036 (+/-0.014) and -0.028 (+/-0.009) for milk, fat, protein yield, percent of fat and protein, respectively.