Sequence analysis and protein interactions of Arabidopsis CIA2 and CIL proteins.
A previous screening of Arabidopsis thaliana for mutants exhibiting dysfunctional chloroplast protein transport identified the chloroplast import apparatus (cia) gene. The cia2 mutant has a pale green phenotype and reduced rate of protein import into chloroplasts, but leaf shape and size are similar to wild-type plants of the same developmental stage. Microarray analysis showed that nuclear CIA2 protein enhances expression of the Toc75, Toc33, CPN10 and cpRPs genes, thereby up-regulating protein import and synthesis efficiency in chloroplasts. CIA2-like (CIL) shares 65% sequence identity to CIA2, suggesting that CIL and CIA2 are homologous proteins in Arabidopsis. Here, we further assess the protein interactions and sequence features of CIA2 and CIL. Subcellular localizations of truncated CIA2 protein fragments in our onion transient assay demonstrate that CIA2 contains two nuclear localization signals (NLS) located at amino acids (aa) 62-65 and 291-308, whereas CIL has only one NLS at aa 47-50. We screened a yeast two-hybrid (Y2H) Arabidopsis cDNA library to search for putative CIA2-interacting proteins and identified 12 nuclear proteins, including itself, CIL, and flowering-control proteins (such as CO, NF-YB1, NF-YC1, NF-YC9 and ABI3). Additional Y2H experiments demonstrate that CIA2 and CIL mainly interact with flowering-control proteins via their N-termini, but preferentially form homo- or hetero-dimers through their C-termini. Moreover, sequence alignment showed that the N-terminal sequences of CIA2, CIL and NF-YA are highly conserved. Therefore, NF-YA in the NF-Y complex could be substituted by CIA2 or CIL. We show that Arabidopsis CIA2 and CIL can interact with CO and NF-Y complex, so not only may they contribute to regulate chloroplast function but also to modulate flower development.


Factors Impacting Physical Therapy Utilization for Patients With Nonspecific Low Back Pain: Retrospective Analysis of a Clinical Data Set.
Factors predicting physical therapy utilization for lower back pain (LBP) remain unclear, limiting the development of value-based initiatives. The purpose of this study was to identify important factors that impact the number of physical therapist visits per episode of care for US adults with nonspecific LBP. This study was a retrospective observational cohort study of a clinical dataset derived from 80 clinics of a single physical therapy provider organization. Research variables were categorized at the individual (patient) level and the organization (therapist, clinic) level. A hierarchical regression model was designed to identify factors influencing the number of physical therapist visits per episode of care. Higher out-of-pocket payments per visit, receipt of "active" physical therapy, longer average visit length, earlier use of physical therapy, and sex of the therapist (male) were found to predict fewer visits per episode of care. Percent change of function, prior receipt of physical therapy by the same provider organization, self-discharge from physical therapy, level of starting function, and therapist certification were found to predict more visits. Of the variance in number of visits, 8.0% was attributable to the health care organization. Individual factors, such as higher out-of-pocket payment, have a significant impact on reducing visits per episode of care and should be considered when developing value-based initiatives to optimize clinical and utilization outcomes. Payers use consumer-directed healthcare to reduce costs by discouraging utilization of low value services and encouraging use of low-cost providers. LBP is a costly condition for which physical therapy is a high-value treatment. This study shows that non-need factors predict the number of physical therapy visits per episode of care for patients with nonspecific LBP. Insurance benefit plans with lower out-of-pocket payments for physical therapy and higher reimbursement for active physical therapist interventions may facilitate appropriate utilization of high-value treatment for LBP.


Suggested components of the curriculum for nurses and midwives to enable them to develop essential knowledge and skills in genetics.
Nurses and midwives need to develop specific knowledge and skills in genetics to enable them to offer appropriate healthcare in a range of non-specialist settings. Studies on the topic indicate that while nurses acknowledged the importance of genetics knowledge to their work, both their knowledge and confidence in using such information are poor. Despite the existence of competence frameworks, it appears that educators have struggled with the need to integrate genetics into nursing and midwifery curricula. An expert workshop on genetics education was held to determine the essential components of genetics knowledge and skills that should be incorporated into the pre-registration nursing curriculum in European countries. In this paper we present the essential topics for nurse and midwife pre-registration education and suggest ways in which genetics might be incorporated into the nursing and midwifery curriculum.


The Mycobacterium avium complex.
Mycobacterium avium complex (MAC) disease emerged early in the epidemic of AIDS as one of the common opportunistic infections afflicting human immunodeficiency virus-infected patients. However, only over the past few years has a consensus developed about its significance to the morbidity and mortality of AIDS. M. avium was well known to mycobacteriologists decades before AIDS, and the MAC was known to cause disease, albeit uncommon, in humans and animals. The early interest in the MAC provided a basis for an explosion of studies over the past 10 years largely in response to the role of the MAC in AIDS opportunistic infection. Molecular techniques have been applied to the epidemiology of MAC disease as well as to a better understanding of the genetics of antimicrobial resistance. The interaction of the MAC with the immune system is complex, and putative MAC virulence factors appear to have a direct effect on the components of cellular immunity, including the regulation of cytokine expression and function. There now is compelling evidence that disseminated MAC disease in humans contributes to both a decrease in the quality of life and survival. Disseminated disease most commonly develops late in the course of AIDS as the CD4 cells are depleted below a critical threshold, but new therapies for prophylaxis and treatment offer considerable promise. These new therapeutic modalities are likely to be useful in the treatment of other forms of MAC disease in patients without AIDS. The laboratory diagnosis of MAC disease has focused on the detection of mycobacteria in the blood and tissues, and although the existing methods are largely adequate, there is need for improvement. Indeed, the successful treatment of MAC disease clearly will require an early and rapid detection of the MAC in clinical specimens long before the establishment of the characteristic overwhelming infection of bone marrow, liver, spleen, and other tissue. Also, a standard method of susceptibility testing is of increasing interest and importance as new effective antimicrobial agents are identified and evaluated. Antimicrobial resistance has already emerged as an important problem, and methods for circumventing resistance that use combination therapies are now being studied.


Lower extremity arterial disease: multidetector CT angiography meta-analysis.
To obtain the best available estimates of the diagnostic performance of multidetector computed tomographic (CT) angiography compared with that of digital subtraction angiography (DSA) in the assessment of symptomatic lower extremity arterial disease and to identify the most important sources of variation in diagnostic performance between studies. Reports of studies published from January 2000 through April 2006 in English, German, French, or Spanish were searched for by using the MEDLINE, EMBASE, and Cochrane databases. Studies were included if they allowed construction of 2 x 2 contingency tables for the detection of stenosis of 50% or greater at multidetector CT angiography compared with that at DSA -- the reference standard -- in patients with claudication or critical ischemia. Two observers extracted data about study design, patient characteristics, arterial tracts, and technical protocols. Random-effects summary receiver operating characteristic analysis was performed to examine the influence of these data on diagnostic performance. Of the 70 studies initially identified, 12 were included in which multidetector CT angiography was used to evaluate 9541 arterial segments in 436 patients. The pooled sensitivity and specificity for detecting a stenosis of at least 50% per segment were 92% (95% confidence interval: 89%, 95%) and 93% (95% confidence interval: 91%, 95%), respectively. Three studies provided data about the diagnostic performance of multidetector CT angiography in subdivisions of the arterial tract. The diagnostic performance of multidetector CT angiography in the infrapopliteal tract was lower than but not significantly different from that in the aortoiliac (P > .11) and femoropopliteal (P > .40) tracts. Regression analysis showed that diagnostic performance was not significantly influenced by differences in study characteristics. Multidetector CT angiography is an accurate diagnostic test in the assessment of arterial disease (> or =50% stenosis) of the entire lower extremity.


Food policy in the Canadian North: Is there a role for country food markets?
Food insecurity is widely reported to be at a crisis level in the Inuit territory of Nunavut, Canada. Various policies, programs, and initiatives have been proposed to tackle the problem, with increasing interest in developing a system of country food markets (CFMs) similar to Greenland. We examine if CFMs offer a feasible, sustainable, and effective model for strengthening food systems in Nunavut, examining the model of Greenland and drawing on semi-structured interviews with key informants (n = 45). The Greenland experience indicates that CFMs can provide access to sufficient, safe, and nutritious food on a regular basis, and can diversify locally available foods. These benefits are transferable to Nunavut, although knowledge gaps, regulatory and institutional conditions, and concerns over how CFMs might affect the cultural basis of food systems, underlies apprehension over their development in the territory. We conclude that Nunavut is not currently in the position to develop CFMs, but the role of such markets in potentially strengthening food systems should not be discounted. Future development would need to solicit community input on CFMs, resolve regulatory issues around wildlife management and harvesting, and study how future risks would affect sustainability and effectiveness.


NBS1 mediates ATR-dependent RPA hyperphosphorylation following replication-fork stall and collapse.
Post-translational phosphorylation of proteins provides a mechanism for cells to switch on or off many diverse processes, including responses to replication stress. Replication-stress-induced phosphorylation enables the rapid activation of numerous proteins involved in DNA replication, DNA repair and cell cycle checkpoints, including replication protein A (RPA). Here, we report that hydroxyurea (HU)-induced RPA phosphorylation requires both NBS1 (NBN) and NBS1 phosphorylation. Transfection of both phosphospecific and nonphosphospecific anti-NBS1 antibodies blocked hyperphosphorylation of RPA in HeLa cells. Nijmegen breakage syndrome (NBS) cells stably transfected with an empty vector or with S343A-NBS1 or S278A/S343A phospho-mutants were unable to hyperphosphorylate RPA in DNA-damage-associated foci following HU treatment. The stable transfection of fully functional NBS1 in NBS cells restored RPA hyperphosphorylation. Retention of ATR on chromatin in both NBS cells and in NBS cells expressing S278A/S343A NBS1 mutants decreased after DNA damage, suggesting that ATR is the kinase responsible for RPA phosphorylation. The importance of RPA hyperphosphorylation is demonstrated by the ability of cells expressing a phospho-mutant form of RPA32 (RPA2) to suppress and delay HU-induced apoptosis. Our findings suggest that RPA hyperphosphorylation requires NBS1 and is important for the cellular response to DNA damage.


Correlation of partition coefficients with antisickling activity of simple alcohols, amides and ureas.
The resultant increase of the solubility of hemoglobin S in the presence of various alcohols, amides, and ureas has been found to correlate with the individual partition coefficients of each agent. Comparisons have been made using the log of the slope of the curves obtained from standard ultracentrifugation gelation assays with the antisickling agents and is expressed in terms of log (slope Cs/[A]) = k log P + k' where Cs is the solubility of HbS in gm/dL, [A] is the concentration of the agent in moles/liter, P is the partition coefficient of A, and k (slope) and k' (y intercept) constants. These results are similar to other structure activity relationships developed by Corwin Hansch and co-workers [5-8, 10]. Such comparisons illustrate in part the quantitative binding capabilities of organic molecules to proteins based upon their inherent hydrophobic character which can be calculated and predicted a priori after a few initial observations. This paper demonstrates that the solubility increase of hemoglobin S by various simple alcohols, amides and ureas results directly from the hydrophobic qualities of each additive in a given series. It is further reasoned that these small molecules interact with hydrophobic areas on hemoglobin S and that this conjugal hydrophobic interaction explains in part their mechanism of action and polymer destabilization. The role of the polar group is also shown to be important for activity.


Drumming in immersive virtual reality: the body shapes the way we play.
It has been shown that it is possible to generate perceptual illusions of ownership in immersive virtual reality (IVR) over a virtual body seen from first person perspective, in other words over a body that visually substitutes the person's real body. This can occur even when the virtual body is quite different in appearance from the person's real body. However, investigation of the psychological, behavioral and attitudinal consequences of such body transformations remains an interesting problem with much to be discovered. Thirty six Caucasian people participated in a between-groups experiment where they played a West-African Djembe hand drum while immersed in IVR and with a virtual body that substituted their own. The virtual hand drum was registered with a physical drum. They were alongside a virtual character that played a drum in a supporting, accompanying role. In a baseline condition participants were represented only by plainly shaded white hands, so that they were able merely to play. In the experimental condition they were represented either by a casually dressed dark-skinned virtual body (Casual Dark-Skinned - CD) or by a formal suited light-skinned body (Formal Light-Skinned - FL). Although participants of both groups experienced a strong body ownership illusion towards the virtual body, only those with the CD representation showed significant increases in their movement patterns for drumming compared to the baseline condition and compared with those embodied in the FL body. Moreover, the stronger the illusion of body ownership in the CD condition, the greater this behavioral change. A path analysis showed that the observed behavioral changes were a function of the strength of the illusion of body ownership towards the virtual body and its perceived appropriateness for the drumming task. These results demonstrate that full body ownership illusions can lead to substantial behavioral and possibly cognitive changes depending on the appearance of the virtual body. This could be important for many applications such as learning, education, training, psychotherapy and rehabilitation using IVR.


Agavins Increase Neurotrophic Factors and Decrease Oxidative Stress in the Brains of High-Fat Diet-Induced Obese Mice.
Fructans obtained from agave, called agavins, have recently shown significant benefits for human health including obesity. Therefore, we evaluated the potential of agavins as neuroprotectors and antioxidants by determining their effect on brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF) as well as oxidative brain damage in of obese mice. Male C57BL/6J mice were fed a high-fat diet (HFD) and treated daily with 5% (HFD/A5) or 10% (HFD/A10) of agavins or a standard diet (SD) for 10 weeks. The levels of BDNF and GDNF were evaluated by ELISA. The oxidative stress was evaluated by lipid peroxidation (TBARS) and carbonyls. SCFAs were also measured with GC-FID. Differences between groups were assessed using ANOVA and by Tukey's test considering p < 0.05. The body weight gain and food intake of mice HFD/A10 group were significantly lower than those in the HFD group. Agavins restored BDNF levels in HFD/A5 group and GDNF levels of HFD/A5 and HFD/A10 groups in cerebellum. Interestingly, agavins decreased TBARS levels in HFD/A5 and HFD/A10 groups in the hippocampus, frontal cortex and cerebellum. Carbonyl levels were also lower in HFD/A5 and HFD/A10 for only the hippocampus and cerebellum. It was also found that agavins enhanced SCFAs production in feces. Agavins may act as bioactive ingredients with antioxidant and protective roles in the brain.


Physicians' attitudes, beliefs and knowledge concerning ovarian cancer.
To determine physicians' attitudes, beliefs and knowledge concerning surgical care of women with ovarian cancer. A survey was created from items generated from the literature, a focus group and individual interviews. The survey was mailed on two occasions to all practicing gynecologists, general surgeons and urologists in Ontario. 701 responses were received (overall response rate: 43.7%); 293 were eligible responses. The responses were analyzed in terms of four determinants of surgical care: knowledge, practice patterns, perceived goals of surgery and barriers to accessing surgical care. These variables would be influenced by the surgeon's specialty, access to an oncologist (medical or gynecologic) at one's facility and distance of one's facility to the nearest cancer center with a gynecologic oncologist. Surgeon's specialty and distance from the cancer center influenced both the intraoperative surgical plan and referral practices. The most important goals of surgery were survival and optimal debulking. The barriers to care included available operating time and surgical beds. We have shown that peer influence has reached a ceiling effect in ovarian cancer and novel approaches are required to ensure appropriate referrals, knowledge transfer and provincial resourcing to expert centers to provide optimal surgical care for women with ovarian cancer.


Glycation of glucagon-like peptide-1(7-36)amide: characterization and impaired action on rat insulin secreting cells.
Glucagon-like peptide-1(7-36)amide(truncated GLP-1, tGLP-1) is a potent insulin releasing hormone of the enteroinsular axis. This study has examined glycation of tGLP-1 and effects of such structural modification on insulin secretion. Monoglycated tGLP-1 (M(r) 3463.8, determined by plasma desorption mass spectrometry) was prepared by incubation with glucose under reducing conditions and purified by reversed-phase high performance liquid chromatography. Automated Edman degradation indicated that tGLP-1 was specifically glycated at the amino terminal His7 site. In extracts from mouse small intestine, glycated tGLP-1 represented approximately 14% of the total tGLP-1 content. Effects of glycated and non-glycated tGLP-1 on insulin secretion were examined using glucose-responsive clonal BRIN-BD11 cells. In acute (20 min) incubations, 10(-9) mol/l tGLP-1 enhanced insulin release by 2.2-fold and 1.5-fold at 5.6 and 11.1 mmol/l glucose, respectively. In contrast, 10(-9) mol/l glycated tGLP-1 failed to stimulate secretion and insulin output was decreased by 34-73% following glycation. At 5.6 mmol/l glucose, non-glycated tGLP-1 (3 x 10(-10) mol/l-10(-8) mol/l) exerted a 2.3-fold to 3.2-fold increase in insulin secretion compared with controls. The effect of glycated tGLP-1 at 10(-9) mol/l and 3 x 10(-9) mol/l was reduced by 51-55% compared with non-glycated peptide, and its insulinotropic action was effectively abolished. These data indicate that when tGLP-1 is glycated at the amino terminal His7, this modification substantially reduces the glucose-dependent insulinotropic action of the peptide.


Multiple-suture nonsyndromic craniosynostosis: early and effective management using endoscopic techniques.
The authors present the results of treating infants with multiple-suture nonsyndromic craniosynostosis in whom the authors used minimally invasive endoscopy-assisted techniques and postoperative cranial molding over an 11-year period. A total of 21 patients who presented with multiple-suture (nonsyndromic) craniosynostosis were treated using minimally invasive endoscopy-assisted craniectomies. Surgery was followed by treatment with custommade cranial orthoses for up to 12 months. A total of 48 sutures were treated. The most common was the coronal suture (38 cases) and this was followed by the sagittal (11 cases), metopic (6 cases), and lambdoid (3 cases) sutures. There were 13 male and 8 female pediatric patients. Their ages ranged between 3 weeks and 9 months (mean 3.2 months, median 2.5 months). The sagittal suture was treated with a wide vertex craniotomy via 2 incisions located behind the anterior fontanel and in front of the lambda. The metopic suture underwent a suturectomy as did the coronal and lambdoid sutures. The mean follow-up duration was 61 months (range 3-135 months). There were no deaths. In patients with bicoronal synostosis, brachycephaly was corrected. Patients presenting with vertical dystopia or nasal deviation had these deformities corrected as well. The mean blood loss was 42 ml (range 10-120 ml). The mean hospital length of stay was 1 day. The intraoperative transfusion rate was 0%. The results indicate that nonsyndromic multiple-suture synostosis can be safely and effectively treated using endoscopic techniques. Early treatment of complex multiple-suture synostosis with endoscopic techniques provides an excellent surgical alternative. The results of the present study indicate marked correction of skull base and craniofacial deformities. Endoscopy provides a safe and effective way to treat these patients.


Childhood Trauma, DNA Methylation of Stress-Related Genes, and Depression: Findings From Two Monozygotic Twin Studies.
DNA methylation has been associated with both early life stress and depression. This study examined the combined association of DNA methylation at multiple CpG probes in five stress-related genes with depressive symptoms and tested whether these genes methylation mediated the association between childhood trauma and depression in two monozygotic (MZ) twin studies. The current analysis comprised 119 MZ twin pairs (84 male pairs [mean = 55 years] and 35 female pairs [mean = 36 years]). Peripheral blood DNA methylation of five stress-related genes (BDNF, NR3C1, SLC6A4, MAOA, and MAOB) was quantified by bisulfite pyrosequencing or 450K BeadChip. We applied generalized Poisson linear-mixed models to examine the association between each single CpG methylation and depressive symptoms. The joint associations of multiple CpGs in a single gene or all five stress-related genes as a pathway were tested by weighted truncated product method. Mediation analysis was conducted to test the potential mediating effect of stress gene methylation on the relationship between childhood trauma and depressive symptoms. Multiple CpG probes showed nominal individual associations, but very few survived multiple testing. Gene-based or gene-set approach, however, revealed significant joint associations of DNA methylation in all five stress-related genes with depressive symptoms in both studies. Moreover, two CpG probes in the BDNF and NR3C1 mediated approximately 20% of the association between childhood trauma and depressive symptoms. DNA methylation at multiple CpG sites are jointly associated with depressive symptoms and partly mediates the association between childhood trauma and depression. Our results highlight the importance of testing the combined effects of multiple CpG loci on complex traits and may unravel a molecular mechanism through which adverse early life experiences are biologically embedded.


Prospective study of pneumonia hospitalizations and mortality of U.S. older people: the role of chronic conditions, health behaviors, and nutritional status.
Mortality and hospitalization rates for pneumonia have increased among older Americans during recent years (1979-86), despite a national commitment to the reduction of premature deaths from pneumonia. A prospective study of deaths and hospitalizations attributable to pneumonia was conducted among 5,474 subjects ages 55 and older who participated in the NHANES I Epidemiologic Followup Study. Prevalent chronic conditions, health behaviors, and nutritional status indicators, measured at baseline, were examined in relation to pneumonia hospitalization and death during 12 years of followup. Mortality and hospitalization rates for pneumonia were higher among men than women, and higher among those ages 65 and older than among those 55-64 of both sexes. Risk of pneumonia death was higher among subjects with a history of congestive heart failure, stroke, cancer, or diabetes. Risk of pneumonia hospitalization was higher among subjects with a history of chronic obstructive pulmonary disease and among men who were current smokers. Daily alcohol consumption did not increase risk of pneumonia in this study population. Four measures of nutritional status were examined taking age, prevalent chronic conditions, and cigarette smoking into account: body mass index, arm muscle area, and serum albumin and hemoglobin levels. Risk of pneumonia death was 2.6 times higher in men in the lowest quartile, compared with men in the highest quartile, of body mass index. Similarly, the risk was 4.5 times higher among men in the lowest quartile of arm muscle area. Risk of death from pneumonia was 3.6 times higher among women in the lowest quartile of serum albumin levels compared with women in the highest quartile. Relative risks for these nutritional status indicators remained elevated after adjusting for age and the medical history risk factors. These risk factors should be taken into account when designing and evaluating pneumonia vaccination trials and community prevention programs.


Interchromophoric Interactions Determine the Maximum Brightness Density in DNA Origami Structures.
An ideal point light source is as small and as bright as possible. For fluorescent point light sources, homogeneity of the light sources is important as well as that the fluorescent units inside the light source maintain their photophysical properties, which is compromised by dye aggregation. Here we propose DNA origami as a rigid scaffold to arrange dye molecules in a dense pixel array with high control of stoichiometry and dye-dye interactions. In order to find the highest labeling density in a DNA origami structure without influencing dye photophysics, we alter the distance of two ATTO647N dyes in single base pair steps and probe the dye-dye interactions on the single-molecule level. For small distances strong quenching in terms of intensity and fluorescence lifetime is observed. With increasing distance, we observe reduced quenching and molecular dynamics. However, energy transfer processes in the weak coupling regime still have a significant impact and can lead to quenching by singlet-dark-state-annihilation. Our study fills a gap of studying the interactions of dyes relevant for superresolution microscopy with dense labeling and for single-molecule biophysics. Incorporating these findings in a 3D DNA origami object will pave the way to bright and homogeneous DNA origami nanobeads.


The Protective Role of Garlic on Allergen-Induced Airway Inflammation in Mice.
Asthma is the most prevalent chronic respiratory disease worldwide. Garlic extracts have long been used as a food source and in traditional medicine. Crude extracts of garlic are used as an anti-inflammatory agent and have been reported to exhibit antiasthmatic properties. However, molecular mechanisms of garlic extracts in the context of antiasthmatic airway inflammation are still unclear. In this study, the antiasthmatic effect of garlic extracts on Th1, Th2, and Th3 cytokine profiles and immunoregulatory mechanism were explored using an animal model of allergic asthma. Garlic extracts significantly reduced total inflammatory cell counts and eosinophil infiltration and decreased the production of Dermatophagoides pteronyssinus IgE in serum and Th1/Th2/Th3 cytokine in bronchoalveolar fluid. Enzyme-linked immunosorbent assay analysis demonstrated that garlic extracts downregulated the levels of cytokines and chemokines, namely Th2-related IL-4, IL-5, and IL-13; but they simultaneously upregulated Th1-related IFN-γ, IL-12, and Th3-related IL-10 and TGF-β expression in BALF. The mechanism may be ascribed to the modulation of Th1-, Th2-, and Th3-related cytokine imbalance.


Comprehensive behavioral analysis of calcium/calmodulin-dependent protein kinase IV knockout mice.
Calcium-calmodulin dependent protein kinase IV (CaMKIV) is a protein kinase that activates the transcription factor CREB, the cyclic AMP-response element binding protein. CREB is a key transcription factor in synaptic plasticity and memory consolidation. To elucidate the behavioral effects of CaMKIV deficiency, we subjected CaMKIV knockout (CaMKIV KO) mice to a battery of behavioral tests. CaMKIV KO had no significant effects on locomotor activity, motor coordination, social interaction, pain sensitivity, prepulse inhibition, attention, or depression-like behavior. Consistent with previous reports, CaMKIV KO mice exhibited impaired retention in a fear conditioning test 28 days after training. In contrast, however, CaMKIV KO mice did not show any testing performance deficits in passive avoidance, one of the most commonly used fear memory paradigms, 28 days after training, suggesting that remote fear memory is intact. CaMKIV KO mice exhibited intact spatial reference memory learning in the Barnes circular maze, and normal spatial working memory in an eight-arm radial maze. CaMKIV KO mice also showed mildly decreased anxiety-like behavior, suggesting that CaMKIV is involved in regulating emotional behavior. These findings indicate that CaMKIV might not be essential for fear memory or spatial memory, although it is possible that the activities of other neural mechanisms or signaling pathways compensate for the CaMKIV deficiency.


MG53 nucleates assembly of cell membrane repair machinery.
Dynamic membrane repair and remodelling is an elemental process that maintains cell integrity and mediates efficient cellular function. Here we report that MG53, a muscle-specific tripartite motif family protein (TRIM72), is a component of the sarcolemmal membrane-repair machinery. MG53 interacts with phosphatidylserine to associate with intracellular vesicles that traffic to and fuse with sarcolemmal membranes. Mice null for MG53 show progressive myopathy and reduced exercise capability, associated with defective membrane-repair capacity. Injury of the sarcolemmal membrane leads to entry of the extracellular oxidative environment and MG53 oligomerization, resulting in recruitment of MG53-containing vesicles to the injury site. After vesicle translocation, entry of extracellular Ca(2+) facilitates vesicle fusion to reseal the membrane. Our data indicate that intracellular vesicle translocation and Ca(2+)-dependent membrane fusion are distinct steps involved in the repair of membrane damage and that MG53 may initiate the assembly of the membrane repair machinery in an oxidation-dependent manner.


Modeling the Development of Goal-Specificity in Mirror Neurons.
Neurophysiological studies have shown that parietal mirror neurons encode not only actions but also the goal of these actions. Although some mirror neurons will fire whenever a certain action is perceived (goal-independently), most will only fire if the motion is perceived as part of an action with a specific goal. This result is important for the action-understanding hypothesis as it provides a potential neurological basis for such a cognitive ability. It is also relevant for the design of artificial cognitive systems, in particular robotic systems that rely on computational models of the mirror system in their interaction with other agents. Yet, to date, no computational model has explicitly addressed the mechanisms that give rise to both goal-specific and goal-independent parietal mirror neurons. In the present paper, we present a computational model based on a self-organizing map, which receives artificial inputs representing information about both the observed or executed actions and the context in which they were executed. We show that the map develops a biologically plausible organization in which goal-specific mirror neurons emerge. We further show that the fundamental cause for both the appearance and the number of goal-specific neurons can be found in geometric relationships between the different inputs to the map. The results are important to the action-understanding hypothesis as they provide a mechanism for the emergence of goal-specific parietal mirror neurons and lead to a number of predictions: (1) Learning of new goals may mostly reassign existing goal-specific neurons rather than recruit new ones; (2) input differences between executed and observed actions can explain observed corresponding differences in the number of goal-specific neurons; and (3) the percentage of goal-specific neurons may differ between motion primitives.


Identification and characterization of a novel human myeloid inhibitory C-type lectin-like receptor (MICL) that is predominantly expressed on granulocytes and monocytes.
Inhibitory and activatory C-type lectin-like receptors play an important role in immunity through the regulation of leukocytes. Here, we report the identification and characterization of a novel myeloid inhibitory C-type lectin-like receptor (MICL) whose expression is primarily restricted to granulocytes and monocytes. This receptor, which contains a single C-type lectin-like domain and a cytoplasmic immunoreceptor tyrosine-based inhibitory motif, is related to LOX-1 (lectin-like receptor for oxidized low density lipoprotein-1) and the beta-glucan receptor (Dectin-1) and is variably spliced and highly N-glycosylated. We demonstrate that it preferentially associates with the signaling phosphatases SHP-1 and SHP-2, but not with SHIP. Novel chimeric analyses with a construct combining MICL and the beta-glucan receptor show that MICL can inhibit cellular activation through its cytoplasmic immunoreceptor tyrosine-based inhibitory motif. These data suggest that MICL is a negative regulator of granulocyte and monocyte function.


1,1,3-Trioxo-2H,4H-thieno[3,4-e][1,2,4]thiadiazine (TTD) derivatives: a new class of nonnucleoside human immunodeficiency virus type 1 (HIV-1) reverse transcriptase inhibitors with anti-HIV-1 activity.
We report the development of a new group of nonnucleoside reverse transcriptase inhibitors (NNRTIs). One of the most active congeners of this series of 1,1,3-trioxo-2H,4H-thieno[3,4-e] [1,2,4]thiadiazine (TTD) derivatives, i.e., 2-(3-fluorobenzyl)-4-cyanomethylen-l,1,3-trioxo-2H,4H- thieno [3,4-e] [1,2,4] thiadiazine) (QM96639) was found to inhibit human immunodeficiency virus (HIV) type 1 [HIV-1 (IIIB)] replication in MT-4 cells at a concentration of 0.09 microM. This compound was toxic for the host cells only at a 1,400-fold higher concentration. The TTD derivatives proved effective against a variety of HIV-1 strains, including those that are resistant to 3'-azido-3'-deoxythymidine (AZT), but not against HIV-2 (ROD) or simian immunodeficiency virus (SIV/ MAC251). HIV-1 strains containing the L100I, K103N, V106A, E138K, Y181C, or Y188H mutations in their reverse transcriptase (RT) displayed reduced sensitivity to the compounds. Their cross-resistance patterns correlated with that of nevirapine. 2-Benzyl-4-cyanomethylen-1,1,3-trioxo-2H,4H-thieno[3,4-e] [1,2,4]thiadiazine (QM96521) enhanced the anti-HIV-1 activity of AZT and didanosine in a subsynergistic manner. HIV-1-resistant virus containing the V179D mutation in the RT was selected after approximately six passages of HIV-1 (IIIB) in CEM cells in the presence of different concentrations of QM96521. From structure-activity relationship analysis of a wide variety of TTD derivatives, a number of restrictions appeared as to the chemical modifications that were compatible with anti-HIV activity. Modelling studies suggest that in contrast to most other NNRTIs, but akin to nevirapine, QM96521 does not act as a hydrogen bond donor in the RT-drug complex.


Haemodynamics of ACTH-induced hypertension in sheep.
1. Administration of adrenocorticotropic hormone (ACTH) to sheep produced increases in mean arterial pressure within 24 h associated with an increase in cardiac output and cardiac rate. Both cardiac output and blood pressure remained elevated over the 5 days of ACTH treatment. 2. Administration of ACTH during beta-adrenoreceptor blockade resulted in an increase in blood pressure without changes in cardiac output at 24 h. 3. Administration of a combined steroid infusion over 5 days produced increases in cardiac output identical with the effects of ACTH but with a substantially smaller effect on blood pressure. 4. These data suggest that the observed changes in cardiac output produced by ACTH treatment may be associated with high blood concentrations of adrenocortical steroids rather than being necessary for the development of the hypertension.


Cancer cell-expressed SLAMF7 is not required for CD47-mediated phagocytosis.
CD47 is a prominent new target in cancer immunotherapy, with antagonistic antibodies currently being evaluated in clinical trials. For effective evaluation of this strategy it is crucial to identify which patients are suited for CD47-targeted therapy. In this respect, expression of the pro-phagocytic signal SLAMF7 on both macrophages and cancer cells was recently reported to be a requisite for CD47 antibody-mediated phagocytosis. Here, we demonstrate that in fact SLAMF7 expression on cancer cells is not required and does not impact on CD47 antibody therapy. Moreover, SLAMF7 also does not impact on phagocytosis induction by CD20 antibody rituximab nor associates with overall survival of Diffuse Large B-Cell Lymphoma patients. In contrast, expression of CD47 negatively impacts on overall and progression free survival. In conclusion, cancer cell expression of SLAMF7 is not required for phagocytosis and, in contrast to CD47 expression, should not be used as selection criterion for CD47-targeted therapy.


Warfarin overdose: a 25-year experience.
Warfarin, a vitamin K antagonist, is widely used for the prophylaxis and treatment of thromboembolic disease. While guidelines exist for management of a supratherapeutic international normalized ratio following therapeutic warfarin use, these guidelines are not designed for management of the acute warfarin overdose. There is a paucity of literature describing the latter. The primary objective of this manuscript is to characterize the coagulopathy and describe the bleeding events that occur after a warfarin overdose. A secondary goal is to describe the amount of vitamin K administered to patients presenting with warfarin overdoses. A retrospective chart review of patients admitted with an acute warfarin overdose at two tertiary care medical centers in the USA was conducted. Clinical characteristics were abstracted, and bleeding categories (major, minor, trivial) were defined a priori. Twenty-three patients were admitted during the time period; males accounted for 15/23 (62.5 %) subjects. The median (interquartile range (IQR)) age was 43 (32-48.5) years. Seventeen subjects received vitamin K, with a median (IQR) dose of 15 (10-50) mg. The maximal total amount of vitamin K administered to a single patient during the index hospitalization was 110 mg. Three bleeding events occurred; one classified as major, and two as minor. All patients made a full recovery. In this case series of acute warfarin overdose, nearly all patients developed a coagulopathy, and nearly three-quarters of patients received vitamin K. Bleeding events occurred in a minority of patients.


Oomycete interactions with plants: infection strategies and resistance principles.
The Oomycota include many economically significant microbial pathogens of crop species. Understanding the mechanisms by which oomycetes infect plants and identifying methods to provide durable resistance are major research goals. Over the last few years, many elicitors that trigger plant immunity have been identified, as well as host genes that mediate susceptibility to oomycete pathogens. The mechanisms behind these processes have subsequently been investigated and many new discoveries made, marking a period of exciting research in the oomycete pathology field. This review provides an introduction to our current knowledge of the pathogenic mechanisms used by oomycetes, including elicitors and effectors, plus an overview of the major principles of host resistance: the established R gene hypothesis and the more recently defined susceptibility (S) gene model. Future directions for development of oomycete-resistant plants are discussed, along with ways that recent discoveries in the field of oomycete-plant interactions are generating novel means of studying how pathogen and symbiont colonizations overlap.


Renewable hydrogen from nonvolatile fuels by reactive flash volatilization.
Droplets of nonvolatile fuels such as soy oil and glucose-water solutions can be flash evaporated by catalytic partial oxidation to produce hydrogen in high yields with a total time in the reactor of less than 50 milliseconds. Pyrolysis, coupled with catalytic oxidation of the fuels and their fragments upon impact with a hot rhodium-cerium catalyst surface, avoids the formation of deactivating carbon layers on the catalyst. The catalytic reactions of these products generate approximately 1 megawatt of heat per square meter, which maintains the catalyst surface above 800 degrees C at high drop impact rates. At these temperatures, heavy fuels can be catalytically transformed directly into hydrogen, carbon monoxide, and other small molecules in very short contact times without the formation of carbon.


NK Cell-Mediated Antitumor Effects of a Folate-Conjugated Immunoglobulin Are Enhanced by Cytokines.
Optimally effective antitumor therapies would not only activate immune effector cells but also engage them at the tumor. Folate conjugated to immunoglobulin (F-IgG) could direct innate immune cells with Fc receptors to folate receptor-expressing cancer cells. F-IgG bound to human KB and HeLa cells, as well as murine L1210JF, a folate receptor (FR)-overexpressing cancer cell line, as determined by flow cytometry. Recognition of F-IgG by natural killer (NK) cell Fc receptors led to phosphorylation of the ERK transcription factor and increased NK cell expression of CD69. Lysis of KB tumor cells by NK cells increased by about 5-fold after treatment with F-IgG, an effect synergistically enhanced by treatment with IL2, IL12, IL15, or IL21 (P< 0.001). F-IgG also enhanced the lysis of chronic lymphocytic leukemia cells by autologous NK cells. NK cells significantly increased production of IFNγ, MIP-1α, and RANTES in response to F-IgG-coated KB target cells in the presence of the NK cell-activating cytokine IL12, and these coculture supernatants induced significant T-cell chemotaxis (P< 0.001). F-IgG-coated targets also stimulated FcR-mediated monocyte effector functions. Studies in a murine leukemia model confirmed the intratumoral localization and antitumor activity of F-IgG, as well as enhancement of its effects by IL12 (P =0.05). The antitumor effect of this combination was dependent on NK cells and led to decreased tumor cell proliferation in vivo Thus, F-IgG can induce an immune response against FR-positive tumor cells that is mediated by NK cells and can be augmented by cytokine therapy.


Contrast Sensitivity on 1/f Noise Is More Greatly Impacted by Older Age for the Fovea Than Parafovea.
Contrast sensitivity changes across the visual field with age and is often measured clinically with various forms of perimetry on plain backgrounds. In daily life, the visual scene is more complicated, and therefore, the standard clinical measures of contrast sensitivity may not predict a patient's visual experience in more natural environments. This study aims to determine whether contrast thresholds in older adults are different from younger adults when measured on a 1/f noise background (a nonuniform background whose spatial frequency content is similar to those present in the natural vision environments). Twenty younger (age range, 20 to 35 years) and 20 older adults (age range, 61 to 79 years) with normal ocular health were recruited. Contrast thresholds were measured for a Gabor patch of 6 cycles per degree (sine wave grating masked by a Gaussian envelope of standard deviation 0.17°) presented on 1/f noise background (root-mean-square contrast, 0.05 and 0.20) that subtended 15° diameter of the central visual field. The stimulus was presented at four eccentricities (0°, 2°, 4°, and 6°) along the 45° meridian in the noise background, and nine contrast levels were tested at each eccentricity. The proportion of correct responses for detecting the target at each eccentricity was obtained, and psychometric functions were fit to estimate the contrast threshold. Older adults demonstrate increased contrast thresholds compared with younger adults. There was an eccentricity-dependent interaction with age, with the difference between groups being highest in the fovea compared with other eccentricities. Performance was similar for the two noise backgrounds tested. Our results revealed a strong eccentricity dependence in performance between older and younger adults, highlighting age-related differences in the contrast detection mechanisms between fovea and parafovea for stimuli presented on nonuniform backgrounds.


Role of temperature on the development of hypoxia in blackwater from grass.
In eastern Australia the development of hypoxic blackwater/floodwater and its detrimental consequences are more common in summer than winter. This study examined the effect of temperature on the development of hypoxic conditions which was determined as biochemical oxygen demand (BOD) in floodwater when pasture grass (a source containing labile organic carbon) was inundated. Labile dissolved organic carbon (DOC) in blackwater is one of the main factors that contribute to the development of hypoxic conditions. Temperature plays a key role on the microbial mineralization of labile DOC and hence the development of hypoxic conditions. Prolonged flooding at different seasons (having different temperatures) was simulated in the laboratory by incubating fresh pasture grass cuttings with river water and soil at three different temperatures (20 °C, 27.5 °C and 35 °C) for 20 days. Although this study shows that elevated ambient temperatures can result in more rapid development of hypoxic conditions during the first week of flood peak, it is evident that blackwater formed at relatively moderate ambient temperatures (e.g. 20 °C) has a similar potential to deoxygenate the receiving water bodies, especially after one week's duration of flood peak.


Predictors of systemic embolism in patients with mitral stenosis. A prospective study.
Most studies of the predictors of systemic embolism in patients with mitral stenosis have been retrospective. To prospectively study factors associated with systemic embolism in mitral stenosis. Prospective cohort study. University-affiliated medical institution with 3000 beds. 534 consecutive patients with a mitral valve area of 2.0 cm2 or less; 132 patients were in sinus rhythm, and 402 were in atrial fibrillation. Nine clinical and 10 echocardiographic variables were assessed for prediction of systemic embolism over a mean (+/- SD) follow-up of 36.9 +/- 22.5 months. Diagnosis of systemic embolism was based on symptoms and signs (sudden onset of peripheral arterial ischemic or neurologic manifestations without prodromes) and on findings on computed tomography, angiography, and surgery. For patients in sinus rhythm, age (relative risk [RR], 1.12 [95% CI, 1.04 to 1.21]), the presence of a left atrial thrombus (RR, 37.1 [CI, 2.82 to 487.8]), mitral valve area (RR, 16.9 [CI, 1.53 to 187.0]), and the presence of significant aortic regurgitation (RR, 22.4 [CI, 2.72 to 184.8]) were positively associated with embolism. For patients in atrial fibrillation, previous embolism (RR, 3.11 [CI, 1.66 to 5.85]) was positively associated with embolism; percutaneous balloon mitral commissurotomy (RR, 0.37 [CI, 0.18 to 0.79]) was a negative predictor. It may be prudent to give anticoagulants not only to patients in atrial fibrillation and patients with previous systemic embolism but also to those showing a left atrial thrombus or significant aortic regurgitation on echocardiography. Early percutaneous balloon mitral commissurotomy may also help prevent systemic embolism in patients with mitral stenosis.


Graphene quantum dots: multifunctional nanoplatforms for anticancer therapy.
Graphene quantum dots, the next generation carbon based nanomaterials, due to their outstanding physical, chemical and biological properties, have shown potential in revolutionizing the future of nanomedicine and biotechnology. Their strong size-dependent photoluminescence (PL) and the presence of reactive groups on the GQD surface, which allow their multimodal conjugation with various functional groups and biologically active molecules, make them ideal candidates for cancer diagnosis and treatment. GQDs have been loaded with drugs and labeled with tumor-targeting ligand units that are able to specifically recognize cancer receptors exposed on the cancer cell surface by generating new therapies that are able to allow a more efficient targeted delivery of anticancer agents while minimizing their distribution in healthy tissues, as well as the development of new imaging agents for the in vitro and in vivo diagnosis of several types of cancer. Here, we review the recent advances in the study of the application of GQDs as nanoplatforms for anticancer therapy, taking into account the methods used for their synthesis and functionalization procedures, which can deeply affect their biocompatibility and their electronic and optical features. The biosafety and toxicity aspects of these nanomaterials at cellular and animal levels, mainly related to their size and the kind and degree of surface functionalization, are also discussed.


Deficiency of a surface membrane glycoprotein (Mo1) in man.
Deficiency of a granulocyte surface glycoprotein of 150,000-D had been associated with defective C3- and IgG-dependent phagocytosis in a patient with recurrent bacterial infections. By using monoclonal antibodies, we found that this patient's granulocytes, monocytes, and null cells were deficient in Mo1 (equivalent to OKM1 and Mac-1), a cell surface molecule consisting of two noncovalently linked glycoproteins of 155,000 and 94,000 D. The 155,000-D subunit is closely associated with the human complement receptor that recognizes C3bi and/or a further degradation product termed C3dg (C3bi receptor); the 94,000-D subunit has been shown to be shared, on normal cells, by two other surface membrane glycoproteins: lymphocyte function-associated antigen-1 (LFA-1) and P-150, 95. Both subunits of Mo1 were deficient on the patient's granulocytes as determined by immunoprecipitation with subunit-specific monoclonal antibodies as well as fluorescence analysis. Mol-deficient monocytes, like granulocytes, had defective C3-and IgG-dependent phagocytosis. Natural killing activity by the patient's peripheral blood leukocytes was normal. Mo1-deficient granulocytes and monocytes rosetted normally with sheep erythrocytes coated with C3bi. This rosetting was totally inhibited by a mixture of anti-Mo1 and anti-C3b (the major fragment of C3) receptor antibodies but not by either antibody alone. Since monoclonal antibodies to the 155,000-D subunit of Mo1 can inhibit C3bi receptor binding, immune phagocytosis, opsonized zymosan-induced degranulation, and superoxide generation by normal phagocytes (functions which are defective in Mo1-deficient cells), it appears likely that Mo1 deficiency may in part underlie the functional aberrations leading to recurrent bacterial infections in man.


Pharmacogenomics of endocrine therapy in breast cancer.
The treatment of breast cancer with selective estrogen receptor modulators such as tamoxifen and with aromatase inhibitors represents a major advance in cancer chemotherapy. However, there are large variations among patients in both the therapeutic efficacy and side effects of these drugs. Pharmacogenomics is the study of the role of inheritance in this variation and genetic variation in tamoxifen response represents one of the most striking examples of the potential clinical importance of pharmacogenomics. Tamoxifen requires "metabolic activation" catalyzed by cytochrome P450 2D6 (CYP2D6) to form hydroxylated metabolites-4-hydroxytamoxifen and endoxifen (N-desmethyl-4-hydroxytamoxifen)-both of which are much more potent than is the parent drug. However, CYP2D6 is genetically polymorphic. Approximately 5-8% of Caucasian subjects are CYP2D6 "poor metabolizers" on a genetic basis and, as a result, are relatively unable to catalyze tamoxifen hydroxylation. These same subjects appear to have poorer outcomes when treated with tamoxifen than do CYP2D6 "extensive metabolizers: These data led the US Food and Drug Administration (FDA) to hold public hearings in 2006 on the inclusion of this pharmacogenomic information in tamoxifen labeling. However, a series of important questions still remains to be addressed with regard to tamoxifen pharmacogenomics. There have also been preliminary attempts to study the pharmacogenomics ofaromatase inhibitors, including the application of a genotype-to-phenotype research strategy designed to explore the nature and extent of common DNA sequence variation in the CYP19 gene that encodes aromatase. Those results-together with our current level of understanding of tamoxifen pharmacogenomics-will be reviewed in this chapter and both will be placed within the context of the overall development of pharmacogenomics.


Design of chimeric expression elements that confer high-level gene activity in chromoplasts.
Non-green plastids, such as chromoplasts, generally have much lower activity of gene expression than chloroplasts in photosynthetically active tissues. Suppression of plastid genes in non-green tissues occurs through a complex interplay of transcriptional and translational control, with the contribution of regulation of transcript abundance versus translational activity being highly variable between genes. Here, we have investigated whether the low expression of the plastid genome in chromoplasts results from inherent limitations in gene expression capacity, or can be overcome by designing appropriate combinations of promoters and translation initiation signals in the 5' untranslated region (5'-UTR). We constructed chimeric expression elements that combine promoters and 5'-UTRs from plastid genes, which are suppressed during chloroplast-to-chromoplast conversion in Solanum lycopersicum (tomato) fruit ripening, either just at the translational level or just at the level of mRNA accumulation. These chimeric expression elements were introduced into the tomato plastid genome by stable chloroplast transformation. We report the identification of promoter-UTR combinations that confer high-level gene expression in chromoplasts of ripe tomato fruits, resulting in the accumulation of reporter protein GFP to up to 1% of total cellular protein. Our work demonstrates that non-green plastids are capable of expressing genes to high levels. Moreover, the chimeric cis-elements for chromoplasts developed here are widely applicable in basic and applied research using transplastomic methods.


In vivo development of daptomycin resistance in vancomycin-susceptible methicillin-resistant Staphylococcus aureus severe infections previously treated with glycopeptides.
Our aim was to describe the clinical and microbiological features of four cases of severe vancomycin-susceptible methicillin-resistant Staphylococcus aureus (MRSA) infections in which the vancomycin non-susceptibility development and daptomycin resistance occurred under therapy with teicoplanin (three cases) and daptomycin switched to vancomycin (one case). Clinical data were retrospectively reviewed. On nine clinical epidemiologically unrelated daptomycin-susceptible (DAP-S) and daptomycin-resistant (DAP-R) MRSA, we performed: (i) DAP-VAN-TEC-CFX-RIF minimum inhibitory concentrations (MICs); (ii) glycopeptide resistance detection (GRD) by δ-hemolysis; (iii) glycopeptide population analysis; (iv) molecular characterization by PFGE-MLST-SCCmec-agr-typing; (v) rpoB and mprF single nucleotide polymorphisms (SNPs); (vi) dltA-mprF-atl-sceD expression by real-time quantitative polymerase chain reaction (qPCR). Three out of the four patients did not survive despite salvage treatment; two died with active MRSA infection and one died because of Stenotrophomonas maltophilia sepsis. The fourth patient, in which a reversion to a DAP-S phenotype occurred, survived with daptomycin plus trimethoprim/sulfamethoxazole and oxacillin treatment, and endovascular device removal. Daptomycin resistance development was preceded by a stable heterogeneous vancomycin-intermediate S. aureus (hVISA) or VISA phenotype acquisition, while in one case, daptomycin resistance was preceded by an unstable daptomycin heteroresistance (hDAP) behavior reverting to DAP-S during vancomycin plus rifampin therapy followed by high doses of daptomycin. All DAP-R strains showed hVISA or DAP-R traits, including mutations and/or up-regulation of genes involved in cell wall turnover and cell membrane perturbation. In our study, daptomycin resistance arose during glycopeptide therapy. The emergence of DAP-R isolates was preceded by a stable VISA or hVISA phenotype or by instability reverting to a DAP-S heteroresistant phenotype. Daptomycin, as first-line therapy for the treatment of severe MRSA infections, should be used at optimal dosage combined with other agents such as beta-lactams, to prevent daptomycin resistance occurrence.


Dying trajectory in the last year of life: does cancer trajectory fit other diseases?
To examine differences in the pattern of functional decline among persons dying of cancer and other leading noncancer causes of death. Mortality followback survey of next of kin listed on death certificate. Probability sample of all deaths in the United States. Next of kin for 3,614 decedents that represented 914,335 deaths. Days of difficulty with activities of daily living and mobility in the last year of life. Relative to other decedents, patients with cancer experienced an increased rate of functional impairment beginning as late as 5 months prior to death. For example, only 13.9% of patients with cancer had difficulty getting out of bed or a chair 1-year prior to death. This increased from 22.2% to 63.0% in the last five months of life. In contrast, decedents from other diseases had higher rates of functional impairment 1 year prior to death (approximately 35% had difficulty getting out of bed or chair) and they manifested a more gradual increase in the level of functional decline (approximately 50% had difficulty getting out of bed). Precipitous functional decline was associated with hospice involvement and dying at home. Persons dying of cancer experienced sharp functional decline in the last months of life whereas other decedents' have a more gradual decline. The more precipitous functional decline was associated with hospice involvement and dying at home.


Characterization of the TLR Family in Branchiostoma lanceolatum and Discovery of a Novel TLR22-Like Involved in dsRNA Recognition in Amphioxus.
Toll-like receptors (TLRs) are important for raising innate immune responses in both invertebrates and vertebrates. Amphioxus belongs to an ancient chordate lineage which shares key features with vertebrates. The genomic research on TLR genes in Branchiostoma floridae and Branchiostoma belcheri reveals the expansion of TLRs in amphioxus. However, the repertoire of TLRs in Branchiostoma lanceolatum has not been studied and the functionality of amphioxus TLRs has not been reported. We have identified from transcriptomic data 30 new putative TLRs in B. lanceolatum and all of them are transcribed in adult amphioxus. Phylogenetic analysis showed that the repertoire of TLRs consists of both non-vertebrate and vertebrate-like TLRs. It also indicated a lineage-specific expansion in orthologous clusters of the vertebrate TLR11 family. We did not detect any representatives of the vertebrate TLR1, TLR3, TLR4, TLR5 and TLR7 families. To gain insight into these TLRs, we studied in depth a particular TLR highly similar to a B. belcheri gene annotated as bbtTLR1. The phylogenetic analysis of this novel BlTLR showed that it clusters with the vertebrate TLR11 family and it might be more related to TLR13 subfamily according to similar domain architecture. Transient and stable expression in HEK293 cells showed that the BlTLR localizes on the plasma membrane, but it did not respond to the most common mammalian TLR ligands. However, when the ectodomain of BlTLR is fused to the TIR domain of human TLR2, the chimeric protein could indeed induce NF-κB transactivation in response to the viral ligand Poly I:C, also indicating that in amphioxus, specific accessory proteins are needed for downstream activation. Based on the phylogenetic, subcellular localization and functional analysis, we propose that the novel BlTLR might be classified as an antiviral receptor sharing at least partly the functions performed by vertebrate TLR22. TLR22 is thought to be viral teleost-specific TLR but here we demonstrate that teleosts and amphioxus TLR22-like probably shared a common ancestor. Additional functional studies with other lancelet TLR genes will enrich our understanding of the immune response in amphioxus and will provide a unique perspective on the evolution of the immune system.


Dietary supplement with fermented soybeans, natto, improved the neurobehavioral deficits after sciatic nerve injury in rats.
Clearance of fibrin and associated inflammatory cytokines by tissue-type plasminogen activator (t-PA) is related to improved regeneration in neurological disorder. The biological activity of fermented soybean (natto) is very similar to that of t-PA. We investigated the effect of the dietary supplement of natto on peripheral nerve regeneration. The peripheral nerve injury was produced by crushing the left sciatic nerve with a vessel clamp in Sprague-Dawley rats. The injured animals were fed orally either with saline or natto (16 mg/day) for seven consecutive days after injury. Increased functional outcome such as sciatic nerve functional index, angle of ankle, compound muscle action potential and conduction latency were observed in natto-treated group. Histological examination demonstrated that natto treatment improved injury-induced vacuole formation, S-100 and vessel immunoreactivities and axon loss. Oral intake of natto prolonged prothrombin time and reduced fibrinogen but did not change activated partial thromboplastin time and bleeding time. Furthermore, natto decreased injury-induced fibrin deposition, indicating a tolerant fibrinolytic activity. The treatment of natto significantly improved injury-induced disruption of blood-nerve barrier and loss of matrix component such as laminin and fibronectin. Sciatic nerve crush injury induced elevation of tumor necrosis factor alpha (TNF-alpha) production and caused apoptosis. The increased production of TNF-alpha and apoptosis were attenuated by natto treatment. These findings indicate that oral intake of natto has the potential to augment regeneration in peripheral nerve injury, possibly mediated by the clearance of fibrin and decreased production of TNF-alpha.


The nonsense-mediated decay pathway maintains synapse architecture and synaptic vesicle cycle efficacy.
A systematic Drosophila forward genetic screen for photoreceptor synaptic transmission mutants identified no-on-and-no-off transient C (nonC) based on loss of retinal synaptic responses to light stimulation. The cloned gene encodes phosphatidylinositol-3-kinase-like kinase (PIKK) Smg1, a regulatory kinase of the nonsense-mediated decay (NMD) pathway. The Smg proteins act in an mRNA quality control surveillance mechanism to selectively degrade transcripts containing premature stop codons, thereby preventing the translation of truncated proteins with dominant-negative or deleterious gain-of-function activities. At the neuromuscular junction (NMJ) synapse, an extended allelic series of Smg1 mutants show impaired structural architecture, with decreased terminal arbor size, branching and synaptic bouton number. Functionally, loss of Smg1 results in a ~50% reduction in basal neurotransmission strength, as well as progressive transmission fatigue and greatly impaired synaptic vesicle recycling during high-frequency stimulation. Mutation of other NMD pathways genes (Upf2 and Smg6) similarly impairs neurotransmission and synaptic vesicle cycling. These findings suggest that the NMD pathway acts to regulate proper mRNA translation to safeguard synapse morphology and maintain the efficacy of synaptic function.


Portuguese Observational Study of Ischaemic Stroke in Patients Medicated with Non-Vitamin K Antagonist Oral Anticoagulants.
Clinical trials and subsequent meta-analyses showed advantages of non-vitamin K antagonists oral anticoagulants (NOACs) over vitamin K antagonists (VKAs) in patients with non-valvular atrial fibrillation. The impact of preadmission anticoagulation in acute ischaemic stroke (AIS) has not been established. To compare functional outcome of patients with AIS with preadmission NOACs vs. VKAs. A retrospective analysis was conducted on consecutive AIS patients under oral anticoagulation (VKAs or NOACs) admitted in 4 Portuguese hospitals within a period of 30 months. Two primary outcomes were defined and compared between VKA and NOAC groups: symptomatic intracerebral hemorrhage transformation (sICH) and modified Rankin Scale (mRS) at 3 months. Four hundred sixty-nine patients were included, of whom 332 (70.8%) were treated with VKA and 137 (29.2%) with NOAC. Patients' median age was 78.0 and 234 (49.9%) were male. NOAC-treated patients had a higher median CHA2DS2-VASc score than those under VKA (5.0 vs. 4.0, p = 0.023). The two primary outcomes showed no statistical differences between the VKAs' group and the NOACs' group (sICH: 5.4 vs. 5.4% [p = 0.911]; mRS at 3 months: 3.0 vs. 3.0 [p = 0.646], respectively). Preadmission anticoagulation with NOACs in AIS has a functional impact similar to that of VKAs.


Implant failure and subsequent treatment: report of a complex case.
In a 40-year-old woman, a central incisor was to be replaced with an implant-supported crown. When the implant was uncovered, the soft tissue around the implant showed an unfavorable profile: The adjacent lateral incisor showed significant loss of periodontal attachment. Subsequent soft tissue and connective tissue grafts failed to correct the problem, and an alternative treatment plan was formulated. Errors in the initial treatment, including incorrect implant position and failure to analyze the cause of the periodontal attachment loss are discussed in detail; the options for further treatment measures are outlined; and the treatment that was subsequently carried out is presented.


Play profiles of children with HIV/Aids: A comparative study.
Despite play being widely used in occupational therapy practice as a means and ends in intervention, no research has explored how the play of children with HIV/AIDS is affected. Therefore, the specific processes through which play can be used in therapy with children with HIV/AIDS remains underexplored, especially in low resourced settings. This study aims to compare the level of playfulness of children with HIV/AIDS to that of age and gender matched typically developing children living in the same low resourced community. The Test of Playfulness (ToP) was used to examine the differences and similarities of play between children aged 4-7 years who are typically developing children (n = 52) and children with HIV/AIDS who receive antiretroviral treatment (n = 44). Each child was video recorded playing with a familiar playmate both indoors and outdoors. The videos were scored by two independent raters who were blinded to all aspects of the study. Scores were subjected to Rasch analysis to calculate interval level measure scores. Independent t-tests were used to compare differences between the means of the groups. Typically developing children scored significantly higher when playing outdoors, as compared to children with HIV [t (94) = 3.57, p = .001]. There was no significant difference between mean measure scores for indoor play when comparing typically developing children and children with HIV. Children who are HIV positive scored significantly lower than typically developing children on items related to the ability to suspend reality and framing in both indoor and outdoor environments. Children with HIV/AIDS exhibited lower levels of playfulness when compared to typically developing children. This study provides insight on the play profiles of children with HIV/AIDS, highlighting play deficits and differences between indoor and outdoor play. Occupational therapy can play a significant role in promoting playfulness in children with HIV/AIDS.


Validation of glypican-3-specific scFv isolated from paired display/secretory yeast display library.
Glypican-3 (GPC3) is a heparan-sulfate proteoglycan frequently expressed on the cell membrane of malignant hepatocytes in hepatocellular carcinoma. The capacity for screening potential antibodies in vitro using human hepatocellular lines is critical to ensure binding to this highly post-translationally modified glycophosphatidylinositiol-linked protein. We hypothesized that we could utilize a recently described paired display/secretory yeast library to isolate human-derived scFv against glypican-3 for potential diagnostic and/or therapeutic application. Using two different biotinylated antigen targets, a synthesized 29mer fragment GPC3(550-558) and a truncated GPC3(368-548) fused with glutathione S-transferase (GST) we enriched the yeast display library to greater than 30% target-specific yeast with both positive selection and depletion of streptavidin- and GST-specific clones. After cloning of scFv cDNA from the enriched sub-library, scFv specificity was validated by ELISA for binding to recombinant protein from prokaryotic and eukaryotic sources and ultimately naturally presented human protein on the cell membrane of human hepatocellular cell lines. Specificity was confirmed using non-expressing cell lines and shRNA knockdown. Ultimately, five unique scFv with affinity EC(50) ranging from 5.0-110.9 nM were identified. Using a paired display/secretory yeast library, five novel and unique scFvs for potential humoral or chimeric therapeutic development in human hepatocellular carcinoma were isolated and characterized.


Atopic dermatitis and food allergy.
In order to determine the importance of food sensitization in the etiopathogenesis of atopic dermatitis, we performed a study on 74 patients who fulfilled a previously suggested diagnosis criteria. Of these patients, 17.5% presented allergic rhinitis and 62.2% had associated bronchial asthma. We found that in 64.9% of the patients there was a food sensitization, with milk (36.5%), egg (35.1%) and fish (21.6%) being the most frequently involved. We also observed that 34% of the patients were sensitized to Dermatophagoides pteronyssinus and 24.3% to pollen. These sensitizations were confirmed by means of skin tests, specific IgE and antigen-specific histamine release test. The patients underwent a 3-year follow-up in order to find out the clinical evolution once the causal food was avoided and/or a symptomatic treatment was prescribed. The group of patients with no food sensitization was significantly different from the group with food sensitization: in the first group only 20% of the patients presented a very good clinical evolution (asymptomatic), while in the second group, in 71.4% of the patients the symptoms completely stopped. Nevertheless, in the first year follow-up, we found no significant differences between the two groups. In conclusion, a diet avoiding the causal food combined with a suitable symptomatic treatment, led to an important remission of the skin manifestations in children diagnosed with atopic dermatitis.


The biobanking research infrastructure BBMRI_CZ: a critical tool to enhance translational cancer research.
We introduce the national research biobanking infrastructure, BBMRI_CZ. The infrastructure has been founded by the Ministry of Education and became a partner of the European biobanking infrastructure BBMRI.eu. It is designed as a network of individual biobanks where each biobank stores samples obtained from associated healthcare providers. The biobanks comprise long term storage (various types of tissues classified by diagnosis, serum at surgery, genomic DNA and RNA) and short term storage (longitudinally sampled patient sera). We discuss the operation workflow of the infrastructure that needs to be the distributed system: transfer of the samples to the biobank needs to be accompanied by extraction of data from the hospital information systems and this data must be stored in a central index serving mainly for sample lookup. Since BBMRI_CZ is designed solely for research purposes, the data is anonymised prior to their integration into the central BBMRI_CZ index. The index is then available for registered researchers to seek for samples of interest and to request the samples from biobank managers. The paper provides an overview of the structure of data stored in the index. We also discuss monitoring system for the biobanks, incorporated to ensure quality of the stored samples.


The extraosseous and intraosseous arterial anatomy of the adult elbow.
We investigated the extraosseous and intraosseous arterial anatomy of the human adult elbow. Twenty-two fresh adult cadaveric upper extremities were studied with a technique of combined India-ink and latex injection followed by chemical débridement. The intraosseous vascularity of twelve extremities was then evaluated with a rapid Spalteholz clearing technique. Our findings demonstrated consistent patterns of extraosseous and intraosseous vascular anatomy, which were organized into three vascular arcades: medial, lateral, and posterior. The medial arcade was formed by the superior and inferior ulnar collateral arteries and the posterior ulnar recurrent artery. The lateral arcade was formed by the radial and middle collateral, radial recurrent, and interosseous recurrent arteries. The posterior arcade was formed by the medial and lateral arcades and the middle collateral artery. The intraosseous circulation of the elbow, which was segmental in organization, appeared to be dependent on the local blood supply. The capitellum and the lateral aspect of the trochlea were supplied by posterior perforating vessels arising from the radial recurrent, radial collateral, and interosseous recurrent arteries. The medial aspect of the trochlea was supplied by a circumferential vascular ring originating from the inferior ulnar collateral artery. Watershed areas were apparent between the blood supplies to the medial and lateral aspects of the distal end of the humerus. The olecranon was richly supplied by vessels coursing along its medial aspect from the posterior ulnar recurrent artery and along its lateral aspect from the interosseous recurrent artery. The radial head had a dual extraosseous blood supply from a single branch of the radial recurrent artery, which supplied the head directly, and from additional vessels from both the radial and the interosseous recurrent artery, which penetrated the capsular insertion at the neck of the radius. Our findings demonstrate that arterial contributions to the intraosseous circulation of the elbow are more specific than previously appreciated. The intraosseous circulation of the elbow is derived mainly from perforating vessels that arise from neighboring extraosseous arteries. These perforating arteries may be damaged by trauma or by extensile dissection during reconstruction of the elbow. An understanding of the extraosseous and intraosseous circulation of the elbow may help to avoid iatrogenic injury to the intraosseous circulation.


Anisotropy in Stable Conformations of Hydroxylate Ions between the {001} and {110} Planes of TiO2 Rutile Crystals for Glycolate, Lactate, and 2-Hydroxybutyrate Ions Studied by Metadynamics Method.
Control over TiO2 rutile crystal growth and morphology using additives is essential for the development of functional materials. Computer simulation studies on the thermodynamically stable conformations of additives at the surfaces of rutile crystals contribute to understanding the mechanisms underlying this control. In this study, a metadynamics method was combined with molecular dynamics simulations to investigate the thermodynamically stable conformations of glycolate, lactate, and 2-hydroxybutyrate ions at the {001} and {110} planes of rutile crystals. Two simple atom-atom distances were selected as collective variables for the metadynamics method. At the {001} plane, a conformation in which the COO- group was oriented toward the surface was found to be the most stable for the lactate and 2-hydroxybutyrate ions, whereas a conformation in which the COO- group was oriented toward water was the most stable for the glycolate ion. At the {110} plane, a conformation in which the COO- group was oriented toward the surface was the most stable for all three hydroxylate ions, and a second most stable conformation was also observed for the lactate ion at positions close to the {110} plane. For all three hydroxylate ions (α-hydroxycarboxylate ions), the stability of the most stable conformation was higher for the {110} plane than for the {001} plane. At both planes, the stability of the most stable conformation was highest for the 2-hydroxybutyrate ion and lowest for the glycolate ion. Supposing that all three hydroxylate ions serve to decrease the surface free energy at the rutile surface and that a more stable conformation at the rutile surface leads to a greater decrease in the surface free energy, the present results partially explain experimentally observed differences in the changes in growth rate and morphology of rutile crystals in the presence of glycolic, lactic, and 2-hydroxybutyric acids.


Mortality in the uninsured compared with that in persons with public and private health insurance.
To compare mortality in persons with employer-provided health insurance, Medicare, Medicaid, military health benefits, other private health insurance, and no health insurance, before and after adjustment for income and employment status. Cohort study using national survey data containing information on social, economic, and demographic factors and health insurance, with deaths identified through matching to the National Death Index resulting in a mortality follow-up period of 5 years. Noninstitutionalized population of the United States. Approximately 150,000 respondents to national surveys conducted by the US Bureau of the Census (Current Population Surveys), aged 25 to 64 years. After adjustment for age and income, persons with Medicare and Medicaid had the highest mortality in comparison with those with employer-provided insurance, with relative risks generally greater than 2. With adjustment for age and income, persons without insurance had higher mortality than those with employer-provided insurance, with relative risks of 1.2 for white men and 1.5 for white women. These relationships held after adjustment for employment status, with the working uninsured showing mortality between 1.2 and 1.3 times higher than that of the working insured. Mortality was higher in those with lower incomes after adjustment for insurance status. Those with annual income of $10,000 or less per year had mortality about two times that of persons with incomes greater than $25,000 per year. Mortality was lowest in employed persons with employer-provided health insurance. The higher mortality in those with public insurance or with no insurance reflects an indeterminate mix of selection on existing health status and access to medical care.


Field study of early implementation of UV sources and their relative effectiveness for public health and safety.
The emergence of COVID-19 and its corresponding public health burden has prompted industries to rapidly implement traditional and novel control strategies to mitigate the likelihood of SARS-CoV-2 transmission, generating a surge of interest and application of ultraviolet germicidal irradiation (UVGI) sources as disinfection systems. With this increased attention the need to evaluate the efficacy and safety of these types of devices is paramount. A field study of the early implementation of UVGI devices was conducted at the Space Needle located in Seattle, Washington. Six devices were evaluated, including four low-pressure (LP) mercury-vapor lamp devices for air and surface sanitation not designed for human exposure and two krypton chloride (KrCl*) excimer lamp devices to be operated on and around humans. Emission spectra and ultraviolet (UV) irradiance at different locations from the UV devices were measured and germicidal effectiveness against SARS-CoV-2 was estimated. The human safety of KrCl* excimer devices was also evaluated based on measured irradiance and estimated exposure durations. Our results show all LP devices emitted UV radiation primarily at 254 nm as expected. Both KrCl* excimers emitted far UVC irradiation at 222 nm as advertised but also emitted at longer, more hazardous wavelengths (228 to 262 nm). All LP devices emitted strong UVC irradiance, which was estimated to achieve three log reduction of SARS-CoV-2 within 10 sec of exposure at reasonable working distances. KrCl* excimers, however, emitted much lower irradiance than needed for effective disinfection of SARS-CoV-2 (>90% inactivation) within the typical exposure times. UV fluence from KrCl* excimer devices for employees was below the American Conference of Governmental Industrial Hygienists (ACGIH) Threshold Limit Values (TLVs) under the reported device usage and work shifts. However, photosensitive individuals, human susceptibility, or exposure to multiple UV sources throughout a worker's day, were not accounted for in this study. Caution should be used when determining the acceptability of UV exposure to workers in this occupational setting and future work should focus on UVGI sources in public settings.


Early Heart Rate Variability and Electroencephalographic Abnormalities in Acutely Brain-Injured Children Who Progress to Brain Death.
Heart rate variability is controlled by the autonomic nervous system. After brain death, this autonomic control stops, and heart rate variability is significantly decreased. However, it is unknown if early changes in heart rate variability are predictive of progression to brain death. We hypothesized that in brain-injured children, lower heart rate variability is an early indicator of autonomic system failure, and it predicts progression to brain death. We additionally explored the association between heart rate variability and markers of brain dysfunction such as electroencephalogram and neurologic examination between brain-injured children who progressed to brain death and those who survived. Retrospective case-control study. PICU, single institution. Children up to 18 years with a Glasgow Coma Scale score of less than 8 admitted between August of 2016 and December of 2017, who had electrocardiographic data available for heart rate variability analysis, were included. patients who died of causes other than brain death. Twenty-three patients met inclusion criteria: six progressed to brain death (cases), and 17 survived (controls). Five-minute electrocardiogram segments were used to estimate heart rate variability in the time domain (SD of normal-normal intervals, root mean square successive differences), frequency domain (low frequency, high frequency, low frequency/high frequency ratio), Poincaré plots, and approximate entropy. None. Patients who progressed to brain death exhibited significantly lower heart rate variability in the time domain, frequency domain, and Poincaré plots (p < 0.01). The odds of death increased with decreasing low frequency (odds ratio, 4.0; 95% CI, 1.2-13.6) and high frequency (odds ratio, 2.5; 95% CI, 1.2-5.4) heart rate variability power (p < 0.03). Heart rate variability was significantly lower in those with discontinuous or attenuated/featureless electroencephalogram versus those with slow/disorganized background (p < 0.03). These results support the concept of autonomic system failure as an early indicator of impending brain death in brain-injured children. Furthermore, decreased heart rate variability is associated with markers of CNS dysfunction such as electroencephalogram abnormalities.


Infection with Pneumocystis carinii is prevalent in healthy Gambian children.
Pneumocystis pneumonia is rarely identified in the many immunosuppressed individuals with acquired immune deficiency syndrome (AIDS) and malnutrition in Africa. To test whether infection with Pneumocystis carinii occurs in the continent we conducted a comparative serological study, measuring by enzyme-linked immunosorbent assay antibodies to the parasite in 150 healthy young individuals from both Britain and the Gambian savanna. The prevalence of significant titres of antibody to P. carinii steadily increased with age and included more than 70% of both populations by 8 years of age. Infection with P. carinii is, therefore, common in the Gambia. Thus opportunistic pneumocystis pneumonia may be an important but largely unrecognized disease in the continent, though its impact is probably diminished by the prevalence of fatal tuberculous infection, particularly in the AIDS population.


Pharmacokinetic and pharmacodynamic studies of acute interaction between warfarin enantiomers and metronidazole in rats.
This investigation was designed to determine if the reported stereoselectivity of the pharmacokinetic interaction between warfarin and metronidazole in humans occurs also in rats and if the potentiation of the anticoagulant effect of warfarin can be ascribed solely to inhibition of warfarin metabolism by metronidazole. Metronidazole had no effect on the serum protein binding of racemic warfarin in vitro over a wide concentration range but decreased the protein binding of R-(+)-warfarin and S-(-)-warfarin in vivo, perhaps indirectly through metabolite(s). Treatment with i.p. metronidazole, 100 mg/kg every 6 hr, decreased the plasma clearance of free warfarin ("intrinsic clearance"). This inhibitory effect was more pronounced with S-(-)-warfarin than with R-(+)-warfarin (congruent to 60 and congruent to 30%, respectively). Metronidazole did not affect plasma prothrombin complex activity in vitro but reduced it in vivo. Metronidazole treatment increased the elimination rate constant for endogenous prothrombin complex activity and decreased the plasma concentration of free R-(+)-warfarin required to decrease prothrombin complex activity synthesis rate to one-half of normal (there were insufficient data to determine the effect of metronidazole on the activity of the other enantiomer). It is concluded that metronidazole preferentially inhibits the metabolism of S-(-)-warfarin in rats as in humans but that (at least in rats) the stereoselectivity is not absolute. Moreover, metronidazole has an indirect inhibitory effect on warfarin protein binding in vivo and also affects the pharmacodynamics of warfarin as well as the blood clotting process per se.


Modulation of corticotropin-releasing hormone type 2 receptor and urocortin 1 and urocortin 2 mRNA expression in the cardiovascular system of prairie voles following acute or chronic stress.
The purpose of this study was to compare the effects of an acute stressor (restraint) versus a chronic stressor (social isolation) on the expression of mRNAs for corticotropin-releasing hormone receptor type 2 (CRH-R2) and urocortin 1 (Ucn 1) and urocortin 2 (Ucn 2) in the cardiovascular system of socially monogamous prairie voles of both sexes. Acute restraint for 1 h was followed by a marked increase in plasma corticosterone, and when the animals were re-paired for 1 day, the increment of corticosterone was normalized. However, following chronic social isolation for 4 weeks, plasma corticosterone did not differ significantly from the levels measured in animals living in pairs. Restraint or isolation significantly decreased CRH-R2 mRNA in ventricle, atria, and aorta; however, when these animals were re-paired for 1 day, the modulation of CRH-R2 mRNA was normalized in restraint but not in isolated animals. Restraint stress increased the Ucn 1 mRNA expression in the heart of female and male prairie voles, and when the animals were re-paired, the modulation of Ucn 1 mRNA expression was normalized. However, chronic isolation showed no effect on cardiac Ucn 1 mRNA expression. Although acute restraint stress produced no effect on the cardiac Ucn 2 mRNA expression, chronic isolation was followed by an increased heart Ucn 2 mRNA expression in both sexes. When the isolated animals were re-paired for 1 day, the cardiac Ucn 2 mRNA expression remained upregulated. The results of the present study reveal that acute restraint as well as social isolation can have significant consequences for the modulation of gene expression for the CRH-R2 and the urocortin peptides in cardiovascular tissue in female and male prairie voles.


Chemical gas-generating nanoparticles for tumor-targeted ultrasound imaging and ultrasound-triggered drug delivery.
Although there is great versatility of ultrasound (US) technologies in the real clinical field, one main technical challenge is the compromising of high quality of echo properties and size engineering of ultrasound contrast agents (UCAs); a high echo property is offset by reducing particle size. Herein, a new strategy for overcoming the dilemma by devising chemical gas (CO2) generating carbonate copolymer nanoparticles (Gas-NPs), which are clearly distinguished from the conventional gas-encapsulated micro-sized UCAs. More importantly, Gas-NPs could be readily engineered to strengthen the desirable in vivo physicochemical properties for nano-sized drug carriers with higher tumor targeting ability, as well as the high quality of echo properties for tumor-targeted US imaging. In tumor-bearing mice, anticancer drug-loaded Gas-NPs showed the desirable theranostic functions for US-triggered drug delivery, even after i.v. injection. In this regard, and as demonstrated in the aforementioned study, our technology could serve a highly effective platform in building theranostic UCAs with great sophistication and therapeutic applicability in tumor-targeted US imaging and US-triggered drug delivery.


The transcriptional repressor DEC2 regulates sleep length in mammals.
Sleep deprivation can impair human health and performance. Habitual total sleep time and homeostatic sleep response to sleep deprivation are quantitative traits in humans. Genetic loci for these traits have been identified in model organisms, but none of these potential animal models have a corresponding human genotype and phenotype. We have identified a mutation in a transcriptional repressor (hDEC2-P385R) that is associated with a human short sleep phenotype. Activity profiles and sleep recordings of transgenic mice carrying this mutation showed increased vigilance time and less sleep time than control mice in a zeitgeber time- and sleep deprivation-dependent manner. These mice represent a model of human sleep homeostasis that provides an opportunity to probe the effect of sleep on human physical and mental health.


Biochemical evidence for a mitochondrial genetic modifier in the phenotypic manifestation of Leber's hereditary optic neuropathy-associated mitochondrial DNA mutation.
Leber's hereditary optic neuropathy (LHON) is the most common mitochondrial disease. Mitochondrial modifiers are proposed to modify the phenotypic expression of primary LHON-associated mitochondrial DNA (mtDNA) mutations. In this study, we demonstrated that the LHON susceptibility allele (m.14502T > C, p. 58I > V) in the ND6 gene modulated the phenotypic expression of primary LHON-associated m.11778G > A mutation. Twenty-two Han Chinese pedigrees carrying m.14502T > C and m.11778G > A mutations exhibited significantly higher penetrance of optic neuropathy than those carrying only m.11778G > A mutation. We performed functional assays using the cybrid cell models, generated by fusing mtDNA-less ρo cells with enucleated cells from LHON patients carrying both m.11778G > A and m.14502T > C mutations, only m.14502T > C or m.11778G > A mutation and a control belonging to the same mtDNA haplogroup. These cybrids cell lines bearing m.14502T > C mutation exhibited mild effects on mitochondrial functions compared with those carrying only m.11778G > A mutation. However, more severe mitochondrial dysfunctions were observed in cell lines bearing both m.14502T > C and m.11778G > A mutations than those carrying only m.11778G > A or m.14502T > C mutation. In particular, the m.14502T > C mutation altered assemble of complex I, thereby aggravating the respiratory phenotypes associated with m.11778G > A mutation, resulted in a more defective complex I. Furthermore, more reductions in the levels of mitochondrial ATP and increasing production of reactive oxygen species were also observed in mutant cells bearing both m.14502T > C and m.11778G > A mutation than those carrying only 11778G > A mutation. Our findings provided new insights into the pathophysiology of LHON that were manifested by interaction between primary and secondary mtDNA mutations.


Molecular prediction of disease risk and severity in a large Dutch Crohn's disease cohort.
Crohn's disease and ulcerative colitis have a complex genetic background. We assessed the risk for both the development and severity of the disease by combining information from genetic variants associated with inflammatory bowel disease (IBD). We studied 2804 patients (1684 with Crohn's disease and 1120 with ulcerative colitis) and 1350 controls from seven university hospitals. Details of the phenotype were available for 1600 patients with Crohn's disease and for 800 with ulcerative colitis. Genetic association for disease susceptibility was tested for the nucleotide-binding and oligomerisation domain 2 gene (NOD2), the IBD5 locus, the Drosophila discs large homologue 5 and autophagy-related 16-like 1 genes (DLG5 and ATG16L1) and the interleukin 23 receptor gene (IL23R). Interaction analysis was performed for Crohn's disease using the most associated single nucleotide polymorphism (SNP) for each locus. Odds ratios were calculated in an ordinal regression analysis with the number of risk alleles as an independent variable to analyse disease development and severity. Association with Crohn's disease was confirmed for NOD2, IBD5, DLG5, ATG16L1 and IL23R. Patients with Crohn's disease carry more risk alleles than controls (p = 3.85 x 10(-22)). Individuals carrying an increasing number of risk alleles have an increasing risk for Crohn's disease, consistent with an independent effects multiplicative model (trend analysis p = 4.25 x 10(-23)). Patients with Crohn's disease with a more severe disease course, operations or an age of onset below 40 years have more risk alleles compared to non-stricturing, non-penetrating behaviour (p = 0.0008), no operations (p = 0.02) or age of onset above 40 years (p = 0.028). Crohn's disease is a multigenic disorder. An increase in the number of risk alleles is associated with an increased risk for the development of Crohn's disease and with a more severe disease course. Combining information from the known common risk polymorphisms may enable clinicians to predict the course of Crohn's disease.