Workplace interventions for smoking cessation.
The workplace has potential as a setting through which large groups of people can be reached to encourage smoking cessation. To categorise workplace interventions for smoking cessation tested in controlled studies and to determine the extent to which they help workers to stop smoking or to reduce tobacco consumption. We searched the Tobacco Addiction Review Group trials register in November 2002, Medline (1966 - November 2002), EMBASE (1985 - November 2002) and PsycINFO (to November 2002). We searched abstracts from international conferences on tobacco and we checked the bibliographies of identified studies and reviews for additional references. We categorised interventions into two groups: a) Interventions aimed at the individual to promote smoking cessation and b) interventions aimed at the workplace as a whole. We applied different inclusion criteria for the different types of study. For interventions aimed at helping individuals to stop smoking, we included only randomised controlled trials allocating individuals, workplaces or companies to intervention or control conditions. For studies of smoking restrictions and bans in the workplace, we also included controlled trials with baseline and post-intervention outcomes and interrupted times series studies. Information relating to the characteristics and content of all kinds of interventions, participants, outcomes and methods of the study was abstracted by one reviewer and checked by two others. Because of heterogeneity in the design and content of the included studies, we did not attempt formal meta-analysis, and evaluated the studies using qualitative narrative synthesis. Workplace interventions aimed at helping individuals to stop smoking included nine studies of group therapy, three studies of individual counselling, eight studies of self-help materials and four studies of nicotine replacement therapy. The results were consistent with those found in other settings. Group programmes, individual counselling and nicotine replacement therapy increased cessation rates in comparison to no treatment or minimal intervention controls. Self-help materials were less effective. Workplace interventions aimed at the workforce as a whole included 13 studies of tobacco bans, two studies of social support, four studies of environmental support, four studies of incentives, six studies of comprehensive (multi-component) programmes and two studies of competitions and recruitment. Tobacco bans decreased cigarette consumption during the working day but their effect on total consumption was less certain. We failed to detect an increase in quit rates from adding social and environmental support to these programmes. There was a lack of evidence that comprehensive programmes reduced the prevalence of smoking. Competitions and incentives increased attempts to stop smoking, though there was less evidence that they increased the rate of actual quitting. We found 1. Strong evidence that interventions directed towards individual smokers increase the likelihood of quitting smoking. These include advice from a health professional, individual and group counselling and pharmacological treatment to overcome nicotine addiction. Self-help interventions are less effective. All these interventions are effective whether offered in the workplace or elsewhere. Although people taking up these interventions are more likely to stop, the absolute numbers who quit are low. 2. Limited evidence that participation in programmes can be increased by competitions and incentives organised by the employer. 3. Consistent evidence that workplace tobacco policies and bans can decrease cigarette consumption during the working day by smokers and exposure of non-smoking employees to environmental tobacco smoke at work, but conflicting evidence about whether they decrease prevalence of smoking or overall consumption of tobacco by smokers. 4. A lack of evidence that comprehensive approaches reduce the prevalence of smoking, despite the strong theoretical rationale for their use. 5. A lack of evidence about the cost-effectiveness of workplace programmes.


Allogeneic compact bone-derived mesenchymal stem cell transplantation increases survival of mice exposed to lethal total body irradiation: a potential immunological mechanism.
Radiation-induced injury after accidental or therapeutic total body exposure to ionizing radiation has serious pathophysiological consequences, and currently no effective therapy exists. This study was designed to investigate whether transplantation of allogeneic murine compact bone derived-mesenchymal stem cells (CB-MSCs) could improve the survival of mice exposed to lethal dosage total body irradiation (TBI), and to explore the potential immunoprotective role of MSCs. BALB/c mice were treated with 8 Gy TBI, and then some were administered CB-MSCs isolated from C57BL/6 mice. Survival rates and body weight were analyzed for 14 days post-irradiation. At three days post-irradiation, we evaluated IFN-γ and IL-4 concentrations; CD4(+)CD25(+)Foxp3(+) regulatory T cell (Treg) percentage; CXCR3, CCR5, and CCR7 expressions on CD3(+) T cells; and splenocyte T-bet and GATA-3 mRNA levels. CB-MSC effects on bone marrow hemopoiesis were assessed via colony-forming unit granulocyte/macrophage (CFU-GM) assay. After lethal TBI, compared to non-transplanted mice, CB-MSC-transplanted mice exhibited significantly increased survival, body weight, and CFU-GM counts of bone marrow cells (P < 0.05), as well as higher Treg percentages, reduced IFN-γ, CXCR3 and CCR5 down-regulation, and CCR7 up-regulation. CB-MSC transplantation suppressed Th1 immunity. Irradiated splenocytes directly suppressed CFU-GM formation from bone marrow cells, and CB-MSC co-culture reversed this inhibition. Allogeneic CB-MSC transplantation attenuated radiation-induced hematopoietic toxicity, and provided immunoprotection by alleviating lymphocyte-mediated CFU-GM inhibition, expanding Tregs, regulating T cell chemokine receptor expressions, and skewing the Th1/Th2 balance toward anti-inflammatory Th2 polarization.


Emotion detection deficits and changes in personality traits linked to loss of white matter integrity in primary progressive aphasia.
Non-cognitive features including personality changes are increasingly recognized in the three PPA variants (semantic-svPPA, non fluent-nfvPPA, and logopenic-lvPPA). However, differences in emotion processing among the PPA variants and its association with white matter tracts are unknown. We compared emotion detection across the three PPA variants and healthy controls (HC), and related them to white matter tract integrity and cortical degeneration. Personality traits in the PPA group were also examined in relation to white matter tracts. Thirty-three patients with svPPA, nfvPPA, lvPPA, and 32 HC underwent neuropsychological assessment, emotion evaluation task (EET), and MRI scan. Patients' study partners were interviewed on the Clinical Dementia Rating Scale (CDR) and completed an interpersonal traits assessment, the Interpersonal Adjective Scale (IAS). Diffusion tensor imaging of uncinate fasciculus (UF), superior longitudinal fasciculus (SLF) and inferior longitudinal fasciculus (ILF), and voxel-based morphometry to derive gray matter volumes for orbitofrontal cortex (OFC), anterior temporal lobe (ATL) regions were performed. In addition, gray matter volumes of white matter tract-associated regions were also calculated: inferior frontal gyrus (IFG), posterior temporal lobe (PTL), inferior parietal lobe (IPL) and occipital lobe (OL). ANCOVA was used to compare EET performance. Partial correlation and multivariate linear regression were conducted to examine association between EET and neuroanatomical regions affected in PPA. All three variants of PPA performed significantly worse than HC on EET, and the svPPA group was least accurate at recognizing emotions. Performance on EET was related to the right UF, SLF, and ILF integrity. Regression analysis revealed EET performance primarily relates to the right UF integrity. The IAS subdomain, cold-hearted, was also associated with right UF integrity. Disease-specific emotion recognition and personality changes occur in the three PPA variants and are likely associated with disease-specific neuroanatomical changes. Loss of white matter integrity contributes as significantly as focal atrophy in behavioral changes in PPA.


Involvement of Arg-328, Arg-334 and Arg-342 of DnaA protein in the functional interaction with acidic phospholipids.
We reported previously that three basic amino acids (Arg-360, Arg-364 and Lys-372) of DnaA protein are essential for its functional interaction with cardiolipin. In this study, we examined the effect of mutation of some basic amino acids in a potential amphipathic helix (from Lys-327 to Ile-345) of DnaA protein on this interaction. ATP binding to the mutant DnaA protein, in which Arg-328, Arg-334 and Arg-342 were changed to acidic amino acids, was less inhibited by cardiolipin than that of the wild-type protein, as was the case for mutant DnaA protein with mutations of Arg-360, Arg-364 and Lys-372. A mutant DnaA protein with mutations of all six basic amino acids showed the most resistance to the inhibition of ATP binding by cardiolipin. These results suggest that Arg-328, Arg-334 and Arg-342, like Arg-360, Arg-364 and Lys-372, are also involved in the functional interaction between DnaA protein and acidic phospholipids.


The PHD3 domain of MLL acts as a CYP33-regulated switch between MLL-mediated activation and repression .
The mixed lineage leukemia (MLL) gene plays a critical role in epigenetic regulation of gene expression and is a frequent target of chromosomal translocations leading to leukemia. MLL plant homeodomain 3 (PHD3) is lost in all MLL translocation products, and reinsertion of PHD3 into MLL fusion proteins abrogates their transforming activity. PHD3 has been shown to interact with the RNA-recognition motif (RRM) domain of human nuclear Cyclophilin33 (CYP33). Here, we show that CYP33 mediates downregulation of the expression of MLL target genes HOXC8, HOXA9, CDKN1B, and C-MYC, in a proline isomerase-dependent manner. This downregulation correlates with the reduction of trimethylated lysine 4 of histone H3 (H3K4me3) and histone H3 acetylation. We have structurally characterized both the PHD3 and CYP33 RRM domains and analyzed their binding to one another. The PHD3 domain binds H3K4me3 (preferentially) and the CYP33 RRM domain at distinct sites. Our binding data show that binding of H3K4me3 to PHD3 and binding of the CYP33 RRM domain to PHD3 are mutually inhibitory, implying that PHD3 is a molecular switch for the transition between activation and repression of target genes. To explore the possible mechanism of CYP33/PHD3-mediated repression, we have analyzed the CYP33 proline isomerase activity on various H3 and H4 peptides and shown selectivity for two sites in H3. Our results provide a possible mechanism for the MLL PHD3 domain to act as a switch between activation and repression.


The microinflammatory state in uremia: causes and potential consequences.
Mortality is markedly elevated in patients with end-stage renal disease. The leading cause of death is cardiovascular disease. Lipoprotein levels are only slightly elevated in dialysis patients, and cardiovascular risk is inversely correlated with serum cholesterol, suggesting that a process other than hyperlipidemia plays a role in the incidence of cardiovascular disease. Hypoalbuminemia, ascribed to malnutrition, has been one of the most powerful risk factors that predict all-cause and cardiovascular mortality in dialysis patients. The presence of inflammation, as evidenced by increased levels of specific cytokines (interleukin-6 and tumor necrosis factor alpha) or acute-phase proteins (C-reactive protein and serum amyloid A), however, has been found to be associated with vascular disease in the general population as well as in dialysis patients. The process of inflammation, also called the acute-phase response, additionally causes loss of muscle mass and changes in plasma composition-decreases in serum albumin, prealbumin, and transferrin levels, also associated with malnutrition. Inflammation alters lipoprotein structure and function as well as endothelial structure and function to favor atherogenesis and increases the concentration of atherogenic proteins in serum, such as fibrinogen and lipoprotein (a). Inflammation in dialysis patients is episodic. The causes are likely to be multifactorial and include vascular access infection, less-than-sterile dialysate, dialysate back leak, and nonbiocompatible membranes in addition to clinically apparent infection. In addition, proinflammatory compounds, such as advanced glycation end products, accumulate in renal failure, and defense mechanisms against oxidative injury are reduced, contributing to inflammation and to its effect on the vascular endothelium.


Do satellites detect trends in surface solar radiation?
Long-term variations in solar radiation at Earth's surface (S) can affect our climate, the hydrological cycle, plant photosynthesis, and solar power. Sustained decreases in S have been widely reported from about the year 1960 to 1990. Here we present an estimate of global temporal variations in S by using the longest available satellite record. We observed an overall increase in S from 1983 to 2001 at a rate of 0.16 watts per square meter (0.10%) per year; this change is a combination of a decrease until about 1990, followed by a sustained increase. The global-scale findings are consistent with recent independent satellite observations but differ in sign and magnitude from previously reported ground observations. Unlike ground stations, satellites can uniformly sample the entire globe.


Intravenous fluid therapy in the perioperative and critical care setting: Executive summary of the International Fluid Academy (IFA).
Intravenous fluid administration should be considered as any other pharmacological prescription. There are three main indications: resuscitation, replacement, and maintenance. Moreover, the impact of fluid administration as drug diluent or to preserve catheter patency, i.e., fluid creep, should also be considered. As for antibiotics, intravenous fluid administration should follow the four Ds: drug, dosing, duration, de-escalation. Among crystalloids, balanced solutions limit acid-base alterations and chloride load and should be preferred, as this likely prevents renal dysfunction. Among colloids, albumin, the only available natural colloid, may have beneficial effects. The last decade has seen growing interest in the potential harms related to fluid overloading. In the perioperative setting, appropriate fluid management that maintains adequate organ perfusion while limiting fluid administration should represent the standard of care. Protocols including a restrictive continuous fluid administration alongside bolus administration to achieve hemodynamic targets have been proposed. A similar approach should be considered also for critically ill patients, in whom increased endothelial permeability makes this strategy more relevant. Active de-escalation protocols may be necessary in a later phase. The R.O.S.E. conceptual model (Resuscitation, Optimization, Stabilization, Evacuation) summarizes accurately a dynamic approach to fluid therapy, maximizing benefits and minimizing harms. Even in specific categories of critically ill patients, i.e., with trauma or burns, fluid therapy should be carefully applied, considering the importance of their specific aims; maintaining peripheral oxygen delivery, while avoiding the consequences of fluid overload.


Sugar-sweetened beverages, weight gain, and incidence of type 2 diabetes in young and middle-aged women.
Sugar-sweetened beverages like soft drinks and fruit punches contain large amounts of readily absorbable sugars and may contribute to weight gain and an increased risk of type 2 diabetes, but these relationships have been minimally addressed in adults. To examine the association between consumption of sugar-sweetened beverages and weight change and risk of type 2 diabetes in women. Prospective cohort analyses conducted from 1991 to 1999 among women in the Nurses' Health Study II. The diabetes analysis included 91,249 women free of diabetes and other major chronic diseases at baseline in 1991. The weight change analysis included 51,603 women for whom complete dietary information and body weight were ascertained in 1991, 1995, and 1999. We identified 741 incident cases of confirmed type 2 diabetes during 716,300 person-years of follow-up. Weight gain and incidence of type 2 diabetes. Those with stable consumption patterns had no difference in weight gain, but weight gain over a 4-year period was highest among women who increased their sugar-sweetened soft drink consumption from 1 or fewer drinks per week to 1 or more drinks per day (multivariate-adjusted means, 4.69 kg for 1991 to 1995 and 4.20 kg for 1995 to 1999) and was smallest among women who decreased their intake (1.34 and 0.15 kg for the 2 periods, respectively) after adjusting for lifestyle and dietary confounders. Increased consumption of fruit punch was also associated with greater weight gain compared with decreased consumption. After adjustment for potential confounders, women consuming 1 or more sugar-sweetened soft drinks per day had a relative risk [RR] of type 2 diabetes of 1.83 (95% confidence interval [CI], 1.42-2.36; P<.001 for trend) compared with those who consumed less than 1 of these beverages per month. Similarly, consumption of fruit punch was associated with increased diabetes risk (RR for > or =1 drink per day compared with <1 drink per month, 2.00; 95% CI, 1.33-3.03; P =.001). Higher consumption of sugar-sweetened beverages is associated with a greater magnitude of weight gain and an increased risk for development of type 2 diabetes in women, possibly by providing excessive calories and large amounts of rapidly absorbable sugars.


Perioperative outcomes and complication predictors associated with open and minimally invasive nephroureterectomy.
Minimally invasive nephroureterectomy (MINU) and open nephroureterectomy (ONU) have similar oncological outcomes for treatment of upper tract urothelial carcinoma (UTUC). We investigated perioperative outcomes and predictors of complications associated with MINU and ONU. Using the National Surgical Quality Improvement Program (NSQIP) database, 912 patients were identified that underwent radical nephroureterectomy for UTUC between 2005 and 2013. Logistic regression and contingency table methods used preoperative covariates to predict rates of major (Clavien-Dindo grade ≥ 3) and 16 common perioperative complications. Additional comparisons between treatment groups were performed using unpaired t-tests, Wilcoxon rank-sum tests, or Fisher's Exact tests. P values were adjusted to maintain an experiment-wise p < 0.05. A total of 625 (69%) and 287 (31%) patients underwent MINU and ONU, respectively. ONU was associated with a higher rate of major complications (OR: 2.5, CI: 1.2-5.1, p < 0.03). The incidence of pulmonary embolism (bias adjusted OR: 24, CI: 1.3-441, p < 0.003), postoperative pneumonia (OR: 4.9, CI: 1.7-16, p < 0.0016), and transfusion (OR: 2.7, CI: 1.8-4.0, p < 0.0001) was higher for ONU compared to MINU. There were no significant differences in the incidence of other complications. MINU took longer on average (median 223 versus 213 mins, p < 0.02). Time to discharge was longer for ONU (median 5 versus 4 days, p < 0.0001). No other covariates were independent predictors of major complications regardless of surgical approach. Occurrence of major complications were higher for ONU compared to MINU. These data suggest that MINU is an acceptable surgical option with lower morbidity compared to ONU for the management of UTUC.


Do e-mail alerts of new research increase knowledge translation? A "Nephrology Now" randomized control trial.
As the volume of medical literature increases exponentially, maintaining current clinical practice is becoming more difficult. Multiple, Internet-based journal clubs and alert services have recently emerged. The purpose of this study is to determine whether the use of the e-mail alert service, Nephrology Now, increases knowledge translation regarding current nephrology literature. Nephrology Now is a nonprofit, monthly e-mail alert service that highlights clinically relevant articles in nephrology. In 2007-2008, the authors randomized 1,683 subscribers into two different groups receiving select intervention articles, and then they used an online survey to assess both groups on their familiarity with the articles and their acquisition of knowledge. Of the randomized subscribers, 803 (47.7%) completed surveys, and the two groups had a similar number of responses (401 and 402, respectively). The authors noted no differences in baseline characteristics between the two groups. Familiarity increased as a result of the Nephrology Now alerts (0.23 ± 0.087 units on a familiarity scale; 95% confidence interval [CI]: 0.06-0.41; P = .007) especially in physicians (multivariate odds ratio 1.83; P = .0002). No detectable improvement in knowledge occurred (0.03 ± 0.083 units on a knowledge scale; 95% CI: -0.13 to 0.20; P = .687). An e-mail alert service of new literature improved a component of knowledge translation--familiarity--but not knowledge acquisition in a large, randomized, international population.


A "poly-portfolio" for secondary prevention: a strategy to reduce subsequent events by up to 97% over five years.
A "polypill" for the primary prevention of cardiovascular disease has been proposed. We estimated the projected benefit of a secondary prevention "poly-portfolio" strategy, including pharmacologic and lifestyle approaches for those with coronary heart disease (CHD) or stroke. Based on recent clinical trial results and clinical guidelines, combinations of a high-dose statin, low to standard doses of antihypertensive therapy, aspirin, omega-3 fish oil, cardiac rehabilitation, and diet were evaluated. Patients with CHD, post-myocardial infarction (MI), or stroke were projected to experience 84%, 91%, and 77% reductions, respectively, in CHD events from a pharmacologic approach. Numbers of those needed to treat (NNT) for 5 years were 9 to 11 to prevent 1 CHD event, and 21 to prevent 1 stroke. Post-MI patients were projected to experience a 93% reduction in the risk of CHD death (NNT 16) from a pharmacologic approach and a 97% reduction in the risk of CHD death (NNT 15) with the addition of lifestyle changes. A secondary prevention polyportfolio holds great promise for reducing the burden of cardiovascular disease in the highest risk patients.


A poxvirus-derived vector that directs high levels of expression of cloned genes in mammalian cells.
High levels of expression of cloned genes have been obtained in mammalian cells by using poxvirus-derived insertion/expression vectors. These vectors employ the cis-acting element (CAE I) that directs the transcription of one of the most strongly expressed genes of cowpox virus. This gene (the 160K gene) encodes the 160-kDa protein that is the major component of the A-type cytoplasmic inclusions. Its counterpart in vaccinia virus (VV) is the 94K gene contained in the HindIII A fragment of the viral DNA. Two insertion vectors have been constructed; each is designed to allow cloned genes to be placed immediately downstream of a modified version of CAE I within a poxvirus genome. One vector, p1200, enables the CAE I-cloned-gene constructs to be inserted into the thymidine kinase gene of VV. This vector was used to create a VV recombinant that directed expression of the chloramphenicol acetyltransferase (CAT) gene. The other vector, p2101, enables the CAE I-cloned-gene constructs to be inserted into the VV 94K gene. The prototype of this vector was used to create a VV recombinant that directed expression of a hybrid CAT-lacZ gene. Infection of cultured human cells with these recombinants led to high levels of synthesis of either the CAT gene product or the CAT-lacZ gene product. Each of these proteins was produced in quantities that were easily detected by Coomassie blue staining of total cell proteins resolved by polyacrylamide gel electrophoresis. We estimate that these vectors are capable of directing the synthesis of milligram amounts of gene product per 10(9) mammalian cells.


Impaired development of Th2 cells in IL-13-deficient mice.
We report that Th2 cell cultures generated using T cells or splenocytes from IL-13-deficient mice produce significantly reduced levels of IL-4, IL-5, and IL-10 compared with wild-type. In contrast, IL-4 and IL-5 production by mast cells stimulated in vitro with PMA, ionomycin, or IgE cross-linking are unaffected. In vitro Th2 cell differentiation cannot be rescued by the addition of exogenous factors, but in vivo antigen challenge and administration of IL-13 can increase Th2-like cytokine responses as can infection with the parasitic nematode Nippostrongylus brasiliensis. IL-13-deficient mice also have lower basal levels of serum IgE and biased antigen-specific immunoglobulin responses. Thus, IL-13 is an important regulator of Th2 commitment and may therefore play a central role in atopy and infectious diseases.


An in vitro investigation into the surface roughness of 2 glazed, unglazed, and refinished ceramic materials.
It is common practice for clinicians to adjust the porcelain surfaces of dental prostheses. The change in surface roughness after different surface finishing techniques has attracted the attention of prosthodontists. The present study was undertaken to investigate the average surface roughness of 2 dental ceramics: IPS Empress 2 (layering glass-ceramic, Ivoclar Vivadent) and In-Ceram/Vitadur Alpha (Vita), either glazed, unglazed, or refinished using different techniques. Disks of 6-mm diameter and 2-mm thickness were prepared from IPS Empress 2 and In-Ceram Alumina/Vitadur Alpha ceramics. Specimens were finished for 1 minute to the various stages provided in the sequence of instruments in the Shofu porcelain veneer kit. The average roughness measurements were taken from each specimen with a surface roughness tester. Significant differences in surface roughness were observed between the ceramics and among the finishing techniques used for both IPS Empress 2 and In-Ceram/Vitadur Alpha. It was found that unglazed IPS Empress 2 is rougher than unglazed In-Ceram/Vitadur Alpha. Regardless of the type of ceramic or pretreatment, any adjusted ceramic restoration should be reglazed or subjected to a finishing sequence that is followed through to a final stage of polishing with diamond paste.


Tracking local anesthetic effects using a novel perceptual reference approach.
Drug effects of loco-regional anesthetics are commonly measured by unidimensional pain rating scales. These scales require subjects to transform their perceptual correlates of stimulus intensities onto a visual, verbal, or numerical construct that uses a unitless cognitive reference frame. The conceptual understanding and execution of this magnitude estimation task may vary among individuals and populations. To circumvent inherent shortcomings of conventional experimental pain scales, this study used a novel perceptual reference approach to track subjective sensory perceptions during onset of an analgesic nerve block. In 34 male subjects, nociceptive electric stimuli of 1-ms duration were repetitively applied to left (target) and right (reference) mandibular canines every 5 s for 600 s, with a side latency of 1 ms. Stimulus strength to the target canine was programmed to evoke a tolerable pain intensity perception and remained constant at this level throughout the experiment. A dose of 0.6 ml of articaine 4% was submucosally injected at the left mental foramen. Subjects then reported drug effects by adjusting the stimulus strength (in milliamperes) to the reference tooth, so that the perceived intensity in the reference tooth was equi-intense to the target tooth. Pain and stimulus perception offsets were indicated by subjects. Thus, the current approach for matching the sensory experience in one anatomic location after regional anesthesia allows detailed tracking of evolving perceptual changes in another location. This novel perceptual reference approach facilitates direct and accurate quantification of analgesic effects with high temporal resolution. We propose using this method for future experimental investigations of analgesic/anesthetic drug efficacy.


Pseudouridine monophosphate glycosidase: a new glycosidase mechanism.
Pseudouridine (Ψ), the most abundant modification in RNA, is synthesized in situ using Ψ synthase. Recently, a pathway for the degradation of Ψ was described [Preumont, A., Snoussi, K., Stroobant, V., Collet, J. F., and Van Schaftingen, E. (2008) J. Biol. Chem. 283, 25238-25246]. In this pathway, Ψ is first converted to Ψ 5'-monophosphate (ΨMP) by Ψ kinase and then ΨMP is degraded by ΨMP glycosidase to uracil and ribose 5-phosphate. ΨMP glycosidase is the first example of a mechanistically characterized enzyme that cleaves a C-C glycosidic bond. Here we report X-ray crystal structures of Escherichia coli ΨMP glycosidase and a complex of the K166A mutant with ΨMP. We also report the structures of a ring-opened ribose 5-phosphate adduct and a ring-opened ribose ΨMP adduct. These structures provide four snapshots along the reaction coordinate. The structural studies suggested that the reaction utilizes a Lys166 adduct during catalysis. Biochemical and mass spectrometry data further confirmed the existence of a lysine adduct. We used site-directed mutagenesis combined with kinetic analysis to identify roles for specific active site residues. Together, these data suggest that ΨMP glycosidase catalyzes the cleavage of the C-C glycosidic bond through a novel ribose ring-opening mechanism.


ICOS co-stimulatory receptor is essential for T-cell activation and function.
T-lymphocyte activation and immune function are regulated by co-stimulatory molecules. CD28, a receptor for B7 gene products, has a chief role in initiating T-cell immune responses. CTLA4, which binds B7 with a higher affinity, is induced after T-cell activation and is involved in downregulating T-cell responses. The inducible co-stimulatory molecule (ICOS), a third member of the CD28/CTLA4 family, is expressed on activated T cells. Its ligand B7H/B7RP-1 is expressed on B cells and in non-immune tissues after injection of lipopolysaccharide into animals. To understand the role of ICOS in T-cell activation and function, we generated and analysed ICOS-deficient mice. Here we show that T-cell activation and proliferation are defective in the absence of ICOS. In addition, ICOS -/- T cells fail to produce interleukin-4 when differentiated in vitro or when primed in vivo. ICOS is required for humoral immune responses after immunization with several antigens. ICOS-/- mice showed greatly enhanced susceptibility to experimental autoimmune encephalomyelitis, indicating that ICOS has a protective role in inflammatory autoimmune diseases.


Expanding and Validating a Typology of Intimate Partner Violence: Intersections of Violence and Control Within Relationships.
This research explored an expanded typology of intimate partner violence (IPV), intersecting violent and controlling behaviors of partners. Secondary data were analyzed ( n = 714). Cluster analyses indicated that elements of IPV (control perpetration, control victimization, violence victimization, and violence perpetration) all clustered in high/low patterns, which intersected to form 10 unique categories of IPV. Support for all of Johnson's categories of IPV was found, but there was also evidence for the three control-related categories: Unidirectional Control, Bidirectional Control, and Control Resistance. This expanded typology can move research, practice, and policies beyond the gender symmetry debate.


Moderation of the Relationship Between Reward Expectancy and Prediction Error-Related Ventral Striatal Reactivity by Anhedonia in Unmedicated Major Depressive Disorder: Findings From the EMBARC Study.
Anhedonia, disrupted reward processing, is a core symptom of major depressive disorder. Recent findings demonstrate altered reward-related ventral striatal reactivity in depressed individuals, but the extent to which this is specific to anhedonia remains poorly understood. The authors examined the effect of anhedonia on reward expectancy (expected outcome value) and prediction error- (discrepancy between expected and actual outcome) related ventral striatal reactivity, as well as the relationship between these measures. A total of 148 unmedicated individuals with major depressive disorder and 31 healthy comparison individuals recruited for the multisite EMBARC (Establishing Moderators and Biosignatures of Antidepressant Response in Clinical Care) study underwent functional MRI during a well-validated reward task. Region of interest and whole-brain data were examined in the first- (N=78) and second- (N=70) recruited cohorts, as well as the total sample, of depressed individuals, and in healthy individuals. Healthy, but not depressed, individuals showed a significant inverse relationship between reward expectancy and prediction error-related right ventral striatal reactivity. Across all participants, and in depressed individuals only, greater anhedonia severity was associated with a reduced reward expectancy-prediction error inverse relationship, even after controlling for other symptoms. The normal reward expectancy and prediction error-related ventral striatal reactivity inverse relationship concords with conditioning models, predicting a shift in ventral striatal responding from reward outcomes to reward cues. This study shows, for the first time, an absence of this relationship in two cohorts of unmedicated depressed individuals and a moderation of this relationship by anhedonia, suggesting reduced reward-contingency learning with greater anhedonia. These findings help elucidate neural mechanisms of anhedonia, as a step toward identifying potential biosignatures of treatment response.


The Hsp90 chaperone machinery: conformational dynamics and regulation by co-chaperones.
Hsp90 is a dimeric molecular chaperone required for the activation and stabilization of numerous client proteins many of which are involved in essential cellular processes like signal transduction pathways. This activation process is regulated by ATP-induced large conformational changes, co-chaperones and posttranslational modifications. For some co-chaperones, a detailed picture on their structures and functions exists, for others their contributions to the Hsp90 system is still unclear. Recent progress on the conformational dynamics of Hsp90 and how co-chaperones affect the Hsp90 chaperone cycle significantly increased our understanding of the gearings of this complex molecular machinery. This article is part of a Special Issue entitled: Heat Shock Protein 90 (Hsp90).


Haemodynamic and biochemical responses to L-arginine and L-lysine infusions in normal subjects: L-arginine-induced vasodilatation cannot be explained by non-specific effects of cationic amino acids.
1. Pharmacological stimulation of the synthesis of nitric oxide (NO) may be important in the prevention or treatment of cardiovascular diseases. 2. There is much discussion as to whether the precursor of NO, L-arginine, is able to stimulate basal endothelial NO production. L-Arginine is known to have vasodilating effects. However, it is not clear whether L-arginine-induced vasodilatation is attributable to an increase in NO production or to other systemic effects of L-arginine. 3. To investigate further the mechanisms of the L-arginine-induced vasodilatation, we compared the responses to L-arginine with those to saline and L-lysine in healthy subjects. L-Lysine is not a substrate for NO synthesis, but shares many of L-arginine's other properties. 4. During L-arginine infusion, blood pressure decreased [systolic blood pressure from 120.2 (SD 8.8) to 117.3 (12.1) mmHg (P = 0.05); diastolic blood pressure from 65.3 (5.9) to 61.6 (7.9) mmHg (P < 0.01)], and heart rate and extracellular fluid volume increased. The total peripheral vascular resistance decreased during L-arginine infusion by 18.0 (11.4)% (P < or = 0.05 compared with baseline and compared with L-lysine infusion). These results indicate vasodilation. No changes were observed during L-lysine and saline infusion. 5. Plasma cyclic GMP (the second messenger for NO) increased during L-arginine but also during L-lysine infusion [from 5.7 (1.2) to 6.8 (1.7) nmol/l (P < 0.01), and from 5.8 (1.8) to 7.0 (2.9) nmol/l (P < 0.05) respectively]. Plasma L-citrulline (a by-product of NO synthesis from L-arginine) increased during L-arginine infusion from 30.6 (7.5) to 47.1 (9.9) mumol/l (P < 0.001), but also during L-lysine infusion from 32.7 (6.5) to 42.0 (8.3) mumol/l (P < 0.001). 6. Plasma electrolytes and atrial natriuretic peptide concentrations responded similarly to L-arginine and L-lysine infusion, indicating similar effects on osmolality, plasma volume expansion and potassium distribution. 7. In conclusion, although L-lysine infusion had effects that were similar to those of L-arginine infusion, no vasodilatation was observed. Therefore, these effects cannot account for the L-arginine-induced vasodilatation. This finding indirectly supports the hypothesis that the vasodilatation during L-arginine infusion might be mediated by an increase in NO synthesis. If so, our data suggest that the presumed markers for NO synthesis, plasma cyclic GMP and L-citrulline concentrations, do not accurately reflect this increase. Instead, the rise in plasma cyclic GMP may be related to the rise in ANP. The rise in L-citrulline may be related to competition with L-arginine for the same cell membrane transport mechanism and to stimulation of the urea cycle.


Cytogenetic and molecular analysis of inv dup(15) chromosomes observed in two patients with autistic disorder and mental retardation.
A variety of distinct phenotypes has been associated with supernumerary inv dup(15) chromosomes. Although different cytogenetic rearrangements have been associated with distinguishable clinical syndromes, precise genotype-phenotype correlations have not been determined. However, the availability of chromosome 15 DNA markers provides a means to characterize inv dup(15) chromosomes in detail to facilitate the determination of specific genotype-phenotype associations. We describe 2 patients with an autistic disorder, mental retardation, developmental delay, seizures, and supernumerary inv dup(15) chromosomes. Conventional and molecular cytogenetic studies confirmed the chromosomal origin of the supernumerary chromosomes and showed that the duplicated region extended to at least band 15q13. An analysis of chromosome 15 microsatellite CA polymorphisms suggested a maternal origin of the inv dup(15) chromosomes and biparental inheritance of the two intact chromosome 15 homologs. The results of this study add to the existing literature which suggests that the clinical phenotype of patients with a supernumerary inv dup(15) chromosome is determined not only by the extent of the duplicated region, but by the dosage of genes located within band 15q13 and the origin of the normal chromosomes 15.


Spirometry: tool for pharmacy practitioners to expand direct patient care services.
To introduce pharmacy practitioners to spirometry testing and provide strategies for integrating this "value-added" tool with other direct patient care pharmacy services. Spirometry literature and resources obtained through search strategies including Ovid, PubMed, and Google Scholar. Pharmacists are distinctive members of the multidisciplinary patient care team and can contribute by performing spirometry services for pulmonary patients. Pharmacists have been largely absent from performing this much needed service, perhaps as a result of lack of training or because this testing may be perceived as irrelevant to the pharmacist scope of practice. However, pharmacists are actively integrated with many aspects of pulmonary patient care, including recommending and adjusting inhaled pharmacological agents, monitoring for potential drug-drug and drug-disease interactions, recommending smoking cessation, assessing patient prescription insurance coverage, and educating patients, caregivers, and health care providers on use of prescribed respiratory delivery devices. Adding quality spirometry services, based on American Thoracic Society guidelines for accuracy, would increase the breadth and depth of services for pharmacy practitioners. Spirometry testing is an added tool for expanding direct patient care pharmacy services. Physician support, appropriate pharmacist training, and understanding of reimbursement of spirometry services are essential in providing quality spirometry testing. Future studies are needed to assess the role of pharmacists in performing spirometry and measuring the performance outcomes of pulmonary patients.


Further characterization of Clostridium perfringens small acid soluble protein-4 (Ssp4) properties and expression.
Clostridium perfringens type A food poisoning (FP) is usually caused by C. perfringens type A strains that carry a chromosomal enterotoxin gene (cpe) and produce spores with exceptional resistance against heat and nitrites. Previous studies showed that the extreme resistance of spores made by most FP strains is mediated, in large part, by a variant of small acid soluble protein 4 (Ssp4) that has Asp at residue 36; in contrast, the sensitive spores made by other C. perfringens type A isolates contain an Ssp4 variant with Gly at residue 36. The current study has further characterized Ssp4 properties and expression. Spores made by cpe-positive type C and D strains were found to contain the Ssp4 variant with Gly at residue 36 and were shown to be heat- and nitrite-sensitive; this finding may help to explain why cpe-positive type C and D isolates rarely cause food poisoning. Saturation mutagenesis indicated that both amino acid size and charge at Ssp4 residue 36 are important for DNA binding and for spore resistance. C. perfringens Ssp2 was shown to bind preferentially to GC-rich DNA on gel-shift assays, while Ssp4 preferred binding to AT-rich DNA sequences. Maximal spore heat and nitrite resistance required production of all four C. perfringens Ssps, indicating that these Ssps act cooperatively to protect the spore's DNA, perhaps by binding to different chromosomal sequences. The Ssp4 variant with Asp at residue 36 was also shown to facilitate exceptional spore survival at freezer and refrigerator temperatures. Finally, Ssp4 expression was shown to be dependent upon Spo0A, a master regulator. Collectively, these results provide additional support for the importance of Ssps, particularly the Ssp4 variant with Asp at residue 36, for the extreme spore resistance phenotype that likely contributes to C. perfringens type A food poisoning transmission.


Impact of VIP and cAMP on the regulation of TNF-alpha and IL-10 production: implications for rheumatoid arthritis.
Vasoactive intestinal peptide (VIP) is an anti-inflammatory immunomodulatory neuropeptide with therapeutic potential demonstrated for collagen-induced arthritis. The aim of this study was to characterise its potential anti-arthritic effect on human monocytes, macrophages, T cells, and rheumatoid arthritis synovial membrane cells. Monocytes, macrophages, and T cells derived from human peripheral blood were treated with VIP and compared with other cAMP-elevating drugs for a range of activating stimuli. Cytokine production was assessed for cell cultures and, in addition, the ability of VIPs to activate cAMP response element binding protein. VIP partially suppressed monocyte- and macrophage-derived tumour necrosis factor alpha (TNF-alpha) with no effect on IL-10, whereas VIP fails to regulate IL-10 and TNF-alpha production by T lymphocytes. No such modulation of cytokine profile was observed for rheumatoid arthritis synovial membrane cells. Elevation of intracellular cAMP, on the other hand, potently suppressed macrophage TNF-alpha production and modulated T-cell response by inhibiting TNF-alpha and IFN-gamma. VIP's lack of effect on IL-10 and its slight effect on TNF-alpha results from cAMP being rapidly degraded as the phosphodiesterase IV inhibitor, rolipram, rescues cAMP-dependent activation of cAMP response element binding protein. Interestingly, macrophages stimulated with phorbol 12-myristate 13-acetate/ionomycin displayed an augmented IL-10 response upon addition of dibutyryl cAMP, with corresponding downregulation in TNF-alpha, suggesting a complex interaction between protein kinase C and protein kinase A in cytokine regulation. In conclusion, VIP may represent an efficaceous anti-arthritic treatment modulating macrophage and T-cell cytokine profiles when used alongside a phosphodiesterase inhibitor.


Change in incidence and aetiology of intracerebral haemorrhage in Oxfordshire, UK, between 1981 and 2006: a population-based study.
UK stroke mortality data suggest that the incidence of haemorrhagic stroke has fallen in the past 20 years, but these data do not include deaths of individuals aged 75 years or over. Trends in the older population might differ, since cause varies with age. Our aim was to investigate changes in the population-based incidence of intracerebral haemorrhage according to age and likely aetiology. We used data from the Oxford Community Stroke Project (OCSP; 1981-86) and the Oxford Vascular Study (OXVASC; 2002-06) to investigate changes in the incidence of intracerebral haemorrhage with time, above and below age 75 years, together with associated risk factors and premorbid medications. Incidences were standardised to the 2001 census population of England and Wales. In the population aged under 75 years the incidence of intracerebral haemorrhage decreased substantially (rate ratio 0.53, 95% CI 0.29-0.95; p=0.03), but the number of cases of intracerebral haemorrhage at all ages were similar in OXVASC and OCSP (52 vs 55 cases) as the proportion of cases occurring at 75 years and over tended to increase (2.0, 0.8-4.6; p=0.09). The incidence of intracerebral haemorrhage associated with premorbid hypertension (blood pressure >or=160/100 mm Hg) fell overall (0.37, 0.20-0.69; p=0.002), but the incidence of intracerebral haemorrhage associated with antithrombotic use was increased (7.4, 1.7-32; p=0.007). Above age 75 years the proportion of cases who were non-hypertensive with lobar bleeds and presumed to have had mainly amyloid-related haemorrhages, also increased (4.0, 1.1-17; p=0.003). There has been a substantial fall in hypertension-associated intracerebral haemorrhage over the past 25 years, but not in the overall number of cases of intracerebral haemorrhage in older age-groups, in part due to a rise in intracerebral haemorrhage associated with antithrombotic use. These trends, along with the expected increase in prevalence of amyloid angiopathy with the ageing population, suggest that, in contrast to projections based on mortality data below age 75 years, absolute number of cases of intracerebral haemorrhage might increase in future.


Prevalence of parasitic contamination of raw vegetables in villages of Qazvin Province, Iran.
We conducted a study to determine the extent of parasitic contamination of raw vegetables distributed by wholesalers in villages of Qazvin Province, Iran. Vegetables collected for the study were leek, parsley, lettuce, coriander, radish, spring onion, tarragon, basil, spearmint, and cress. Samples of each vegetable were collected at monthly intervals for 6 months from May to October 2008, yielding a total of 654 samples. Samples of each vegetable collected at each monthly interval were selected randomly and allocated to be washed or to remain unwashed before being analyzed for parasitic contamination. Of the 654 samples collected over the study period, 218 samples were analyzed for parasitic contamination without washing and 436 were washed before being analyzed, with the wash procedure consisting of submersion in a solution of 200 ppm of calcium hypochlorite and rinsing in an automated vegetable washer for 10 minutes. The samples were analyzed according to the method used by the U.S. Food and Drug Administration for identifying parasitic contamination of raw fruits and vegetables, with specimens weighing 200 g each being prepared from the vegetable samples, followed by sonication of each specimen for 10 minutes in 1.5 L of detergent wash solution, collection and centrifugation of a 50-mL aliquot of the wash solution for 15 minutes at 1500 g, and examination of the resulting sediment for parasites by light microscopy. No parasitic contamination was found on any of the 436 washed samples of vegetables. Of the 218 unwashed samples of vegetables, 82 (37.6%) were found to be contaminated with parasites, of which 69 (31.6%) were contaminated with metazoa (helminth eggs in 9.6% and rhabditoid larvae in 22.0%), and 13 (6.0%) were contaminated with protozoa (Entamoeba coli cysts in 2.8%, Entamoeba histolytica/E. dispar cysts in 1.4%, and Giardia lamblia cysts in 1.8%). The difference in parasitic contamination of the washed and unwashed groups of vegetables was significant at p < 0.05. Helminth eggs detected in unwashed samples included those of nematodes (Ascaris lumbricoides [2.3%], Trichuris trichiura [0.9%], and Trichostrongylus spp. [2.8%]), trematodes (Dicrocoelium dendriticum [1.4%]), and cestodes (Taenia spp. [1.8%] and Hymenolepis nana [0.5%]). The eggs recovered in greatest number were those of nematodes (n = 13), followed by those of cestodes (n = 5) and trematodes (n = 3). No helminth eggs were found on unwashed samples of spearmint, tarragon, coriander, or radish. The percent parasitic contamination of unwashed samples was highest for leek (66.7%) and lowest for radish (20.7%). The highest percent parasitic contamination was with rhabditoid larvae (22.0%) and the lowest with H. nana eggs (0.5%). These findings emphasize the importance of properly washing and disinfecting raw vegetables before they are consumed.


Up-regulation of vascular endothelial growth factor expression in a rat glioma is conferred by two distinct hypoxia-driven mechanisms.
Up-regulation of vascular endothelial growth factor (VEGF) expression is a major event leading to neovascularization in malignant gliomas. Hypoxia is believed to be the crucial environmental stimulus for this up-regulation. To critically assess this hypothesis, we asked whether the mechanisms defined previously for hypoxia-induced VEGF expression in vitro are similarly involved and sufficient for up-regulation of VEGF gene expression in vivo, using a lacZ reporter gene under the control of VEGF regulatory sequences in an experimental glioma model. Inclusion of the binding site for hypoxia-inducible factor 1 (HIF 1) in the 5' regulatory sequences used in the hybrid gene produced weak beta-galactosidase staining in a special tumor cell subtype, the so-called perinecrotic palisading (PNP) cells that flank necrotic regions within the tumor. Deletion of the HIF 1 binding site abolished reporter gene expression in the PNP cells, indicating that transcriptional activation of VEGF expression in gliomas is mediated by HIF 1. Inclusion of 3' untranslated sequences from the VEGF gene in the reporter constructs resulted in an increased beta-galactosidase staining in the PNP cells, suggesting that mRNA stabilization also contributes to VEGF up-regulation in glioblastoma cells growing as solid tumors. Combination of the 5' flanking region including the HIF 1 site along with 3' untranslated sequences produced increased levels of beta-galactosidase expression in PNP cells. EF 5 immunostaining for regions of low oxygen partial pressure covered the same PNP cells that were stained for beta-galactosidase. Collectively, the data provide experimental evidence that VEGF gene expression is activated in a distinct tumor cell subpopulation, the perinecrotic palisading cells of gliomas, by two distinct hypoxia-driven regulatory mechanisms.


Dynamics of attention during the initiation of smooth pursuit eye movements.
Many studies indicate that saccades are necessarily preceded by a shift of attention to the target location. There is no direct evidence for the same coupling during smooth pursuit. If smooth pursuit and attention were coupled, pursuit onset should be delayed whenever attention is focused on a stationary, non-target location. To test this hypothesis, observers were instructed to shift their attention to a peripheral location according to a location cue (Experiments 1 and 2) or a symbolic cue (Experiment 3) around the time of smooth pursuit initiation. Attending to static targets had only negligible effects on smooth pursuit latencies and the early open-loop response but lowered pursuit velocity substantially about the onset of closed-loop pursuit. Around this time, eye velocity reflected the competition between the to-be-tracked and to-be-attended object motion, entailing a reduction of eye velocity by 50% compared to the single task condition. The precise time course of attentional modulation of smooth pursuit initiation was at odds with the idea that an attention shift must precede any voluntary eye movement. Finally, the initial catch-up saccades were strongly delayed with attention diverted from the pursuit target. Implications for models of target selection for pursuit and saccades are discussed.


Low-dose hypersensitivity in Chinese hamster V79 cells targeted with counted protons using a charged-particle microbeam.
The Gray Laboratory charged-particle microbeam has been used to assess the clonogenic ability of Chinese hamster V79 cells after irradiation of their nuclei with a precisely defined number of protons with energies of 1.0 and 3.2 MeV. The microbeam uses a 1-microm silica capillary collimator to deliver protons to subcellular targets with high accuracy. The detection system is based on a miniature photomultiplier tube positioned above the cell dish, which detects the photons generated by the passage of the charged particles through an 18-microm-thick scintillator placed below the cells. With this system, a detection efficiency of greater than 99% is achieved. The cells are plated on specially designed dishes (3-microm-thick Mylar base), and the nuclei are identified by fluorescence microscopy. After an incubation period of 3 days, the cells are revisited individually to assess the formation of colonies from the surviving cells. For each energy investigated, the survival curve obtained for the microbeam shows a significant deviation below 1 Gy from a response extrapolated using the LQ model for the survival data above 1 Gy. The data are well fitted by a model that supports the hypothesis that radioresistance is induced by low-dose hypersensitivity. These studies demonstrate the potential of the microbeam for performing studies of the effects of single charged particles on cells in vitro. The hypersensitive responses observed are comparable with those reported by others using different radiations and techniques.


Impact of orbitofrontal lesions on electrophysiological signals in a stop signal task.
Behavioral inhibition and performance monitoring are critical cognitive functions supported by distributed neural networks including the pFC. We examined neurophysiological correlates of motor response inhibition and action monitoring in patients with focal orbitofrontal (OFC) lesions (n = 12) after resection of a primary intracranial tumor or contusion because of traumatic brain injury. Healthy participants served as controls (n = 14). Participants performed a visual stop signal task. We analyzed behavioral performance as well as event-related brain potentials and oscillations. Inhibition difficulty was adjusted individually to yield an equal amount of successful inhibitions across participants. RTs of patients and controls did not differ significantly in go trials or in failed stop trials, and no differences were observed in estimated stop signal RT. However, electrophysiological response patterns during task performance distinguished the groups. Patients with OFC lesions had enhanced P3 amplitudes to congruent condition go signals and to stop signals. In stop trials, patients had attenuated N2 and error-related negativity, but enhanced error positivity. Patients also showed enhanced and prolonged post-error beta band increases for stop errors. This effect was particularly evident in patients whose lesion extended to the subgenual cingulate cortex. In summary, although response inhibition was not impaired, the diminished stop N2 and ERN support a critical role of the OFC in action monitoring. Moreover, the increased stop P3, error positivity, and post-error beta response indicate that OFC injury affected action outcome evaluation and support the notion that the OFC is relevant for the processing of abstract reinforcers such as performing correctly in the task.


Enhancing versus suppressive effects of stress on immune function: implications for immunoprotection and immunopathology.
Stress is known to suppress immune function and increase susceptibility to infections and cancer. Paradoxically, stress is also known to exacerbate asthma, and allergic, autoimmune and inflammatory diseases, although such diseases should be ameliorated by immunosuppression. Moreover, the short-term fight-or-flight stress response is one of nature's fundamental defense mechanisms that enables the cardiovascular and musculoskeletal systems to promote survival, and it is unlikely that this response would suppress immune function at a time when it is most required for survival (e.g. in response to wounding and infection by a predator or aggressor). These observations suggest that stress may suppress immune function under some conditions while enhancing it under others. The effects of stress are likely to be beneficial or harmful depending on the type (immunoprotective, immunoregulatory/inhibitory, or immunopathological) of immune response that is affected. Studies have shown that several critical factors influence the direction (enhancing vs. suppressive) of the effects of stress or stress hormones on immune function: (1) Duration (acute vs. chronic) of stress: Acute or short-term stress experienced at the time of immune activation can enhance innate and adaptive immune responses. Chronic or long-term stress can suppress immunity by decreasing immune cell numbers and function and/or increasing active immunosuppressive mechanisms (e.g. regulatory T cells). Chronic stress can also dysregulate immune function by promoting proinflammatory and type-2 cytokine-driven responses. (2) Effects of stress on leukocyte distribution: Compartments that are enriched with immune cells during acute stress show immunoenhancement, while those that are depleted of leukocytes, show immunosuppression. (3) The differential effects of physiologic versus pharmacologic concentrations of glucocorticoids, and the differential effects of endogenous versus synthetic glucocorticoids: Endogenous hormones in physiological concentrations can have immunoenhancing effects. Endogenous hormones at pharmacologic concentrations, and synthetic hormones, are immunosuppressive. (4) The timing of stressor or stress hormone exposure relative to the time of activation and time course of the immune response: Immunoenhancement is observed when acute stress is experienced at early stages of immune activation, while immunosuppression may be observed at late stages of the immune response. We propose that it is important to study and, if possible, to clinically harness the immunoenhancing effects of the acute stress response, that evolution has finely sculpted as a survival mechanism, just as we study its maladaptive ramifications (chronic stress) that evolution has yet to resolve. In view of the ubiquitous nature of stress and its significant effects on immunoprotection as well as immunopathology, it is important to further elucidate the mechanisms mediating stress-immune interactions and to meaningfully translate findings from bench to bedside.


Risk factors for dental caries in small rural and regional Australian communities.
Dental decay (caries) can cause pain, infection and tooth loss, negatively affecting eating, speaking and general health. People living in rural and regional Australian communities have more caries, more severe caries and more untreated caries than those in the city. The unique environmental conditions and population groups in these communities may contribute to the higher caries burden. In particular, some towns lack community water fluoridation, and some have a high proportion of Aboriginal people, who have significantly worse oral health than their non-Aboriginal counterparts. Because of these and other unique circumstances, mainstream research on caries risk factors may not apply in these settings. This study aimed to gather contemporary oral health data from small rural or regional Australian communities, and investigate caries risk factors in these communities. A cross-sectional survey consisting of a standardized dental examination and questionnaire was used to measure the oral health of 434 children (32% Aboriginal) aged 3-12 years in three small rural or regional areas. Oral health was determined as the deciduous and permanent decayed, missing and filled teeth (dmft/DMFT), and the proportion of children without caries. Risk factors were investigated by logistic regression. The dmft/DMFT for children in this study was 1.5 for 5-6 year olds and 1.0 for 11-12 year olds (index groups reported). Independent predictors of having caries (Yes/No) were age group, holding a concession card (OR=2.45, 95%CI=1.58-3.80) and tooth-brushing less than twice per day (OR=2.11, 95% CI=1.34-3.34). Aboriginal status also became a significant variable under sensitivity analyses (OR 1.9, CI 1.12-3.24) when the tooth-brushing variable was removed. Gender, water fluoridation and parental education were not significant predictors of caries in these communities. The rural/remote children in this study had worse oral health than either state or national average in both the 5-6 year old and 11-12 year age group. Socioeconomic status, tooth-brushing and Aboriginal status were significantly associated with caries in these communities. To close the substantial gap in oral health outcomes between rural and metropolitan residents, approaches that target rural areas, Aboriginal people and those from low socioeconomic backgrounds are needed.


Total synthesis of the reported structure of ceanothine D via a novel macrocyclization strategy.
The first total synthesis of the reported structure of ceanothine D, a cyclopeptide alkaloid found in red root, was achieved using a highly convergent synthetic strategy. Highlights of the synthesis include the first concomitant macrocyclization and formation of the unique chiral tertiary alkyl-aryl ether bond with complete regio- and stereo-control in the presence of a sensitive Z-enamide moiety to access the strained para-cyclophane present in its structure. This synthetic strategy may be broadly applicable in the generation of other structurally similar cyclopeptide alkaloids, enabling further biological and chemical investigations.


Cluster analysis of fMRI data using dendrogram sharpening.
The major disadvantage of hierarchical clustering in fMRI data analysis is that an appropriate clustering threshold needs to be specified. Upon grouping data into a hierarchical tree, clusters are identified either by specifying their number or by choosing an appropriate inconsistency coefficient. Since the number of clusters present in the data is not known beforehand, even a slight variation of the inconsistency coefficient can significantly affect the results. To address these limitations, the dendrogram sharpening method, combined with a hierarchical clustering algorithm, is used in this work to identify modality regions, which are, in essence, areas of activation in the human brain during an fMRI experiment. The objective of the algorithm is to remove data from the low-density regions in order to obtain a clearer representation of the data structure. Once cluster cores are identified, the classification algorithm is run on voxels, set aside during sharpening, attempting to reassign them to the detected groups. When applied to a paced motor paradigm, task-related activations in the motor cortex are detected. In order to evaluate the performance of the algorithm, the obtained clusters are compared to standard activation maps where the expected hemodynamic response function is specified as a regressor. The obtained patterns of both methods have a high concordance (correlation coefficient = 0.91). Furthermore, the dependence of the clustering results on the sharpening parameters is investigated and recommendations on the appropriate choice of these variables are offered. Hum. Brain Mapping 20:201-219, 2003.


Impacts of climate variability and adaptation strategies on crop yields and soil organic carbon in the US Midwest.
Climate change is likely to increase the frequency of drought and more extreme precipitation events. The objectives of this study were i) to assess the impact of extended drought followed by heavy precipitation events on yield and soil organic carbon (SOC) under historical and future climate, and ii) to evaluate the effectiveness of climate adaptation strategies (no-tillage and new cultivars) in mitigating impacts of increased frequencies of extreme events and warming. We used the validated SALUS crop model to simulate long-term maize and wheat yield and SOC changes of maize-soybean-wheat rotation cropping systems in the northern Midwest USA under conventional tillage and no-till for three climate change scenarios (one historical and two projected climates under the Representative Concentration Path (RCP) 4.5 and RCP6) and two precipitation changes (extreme precipitation occurring early or late season). Extended drought events caused additional yield reduction when they occurred later in the season (10-22% for maize and 5-13% for wheat) rather than in early season (5-17% for maize and 2-18% for wheat). We found maize grain yield declined under the projected climates, whereas wheat grain yield increased. No-tillage is able to reduce yield loss compared to conventional tillage and increased SOC levels (1.4-2.0 t/ha under the three climates), but could not reverse the adverse impact of climate change, unless early and new improved maize cultivars are introduced to increase yield and SOC under climate change. This study demonstrated the need to consider extreme weather events, particularly drought and extreme precipitation events, in climate impact assessment on crop yield and adaptation through no-tillage and new genetics reduces yield losses.


Hepatitis C virus envelope glycoprotein E2 glycans modulate entry, CD81 binding, and neutralization.
Hepatitis C virus (HCV) is a major human pathogen that causes serious liver disease, including cirrhosis and hepatocellular carcinoma. The primary target cells of HCV are hepatocytes, and entry is restricted by interactions of the envelope glycoproteins, E1 and E2, with cellular receptors. E1 and E2 form noncovalently linked heterodimers and are heavily glycosylated. Glycans contribute to protein folding and transport as well as protein function. In addition, glycans associated with viral envelopes mask important functional domains from the immune system and attenuate viral immunogenicity. Here, we explored the role of N- and O-linked glycans on E2, which is the receptor binding subunit of the HCV envelope. We identified a number of glycans that are critical for viral entry. Importantly, we showed that the removal of several glycans significantly increased the inhibition of entry by sera from HCV-positive individuals. Only some of the glycans that affected entry and neutralization were also important for CD81 binding. Our results show that HCV envelope-associated glycans play a crucial role in masking functionally important regions of E2 and suggest a new strategy for eliciting highly neutralizing antibodies against this virus.


Millennial-scale temperature change velocity in the continental northern Neotropics.
Climate has been inherently linked to global diversity patterns, and yet no empirical data are available to put modern climate change into a millennial-scale context. High tropical species diversity has been linked to slow rates of climate change during the Quaternary, an assumption that lacks an empirical foundation. Thus, there is the need for quantifying the velocity at which the bioclimatic space changed during the Quaternary in the tropics. Here we present rates of climate change for the late Pleistocene and Holocene from Mexico and Guatemala. An extensive modern pollen survey and fossil pollen data from two long sedimentary records (30,000 and 86,000 years for highlands and lowlands, respectively) were used to estimate past temperatures. Derived temperature profiles show a parallel long-term trend and a similar cooling during the Last Glacial Maximum in the Guatemalan lowlands and the Mexican highlands. Temperature estimates and digital elevation models were used to calculate the velocity of isotherm displacement (temperature change velocity) for the time period contained in each record. Our analyses showed that temperature change velocities in Mesoamerica during the late Quaternary were at least four times slower than values reported for the last 50 years, but also at least twice as fast as those obtained from recent models. Our data demonstrate that, given extremely high temperature change velocities, species survival must have relied on either microrefugial populations or persistence of suppressed individuals. Contrary to the usual expectation of stable climates being associated with high diversity, our results suggest that Quaternary tropical diversity was probably maintained by centennial-scale oscillatory climatic variability that forestalled competitive exclusion. As humans have simplified modern landscapes, thereby removing potential microrefugia, and climate change is occurring monotonically at a very high velocity, extinction risk for tropical species is higher than at any time in the last 86,000 years.


Long-Term Health Outcomes and Health-Related Quality of Life in Adolescents from a Cohort of Extremely Premature Infants Born at Less Than 27 Weeks of Gestation in Northern Germany.
Background Little is known about the psychosocial development and health-related quality of life (HRQOL) of extremely preterm infants once they are adolescents. Methods The regional population-based study cohort included 90 extremely premature infants (< 27+0 gestational weeks) born between January 1997 and December 1999 in the German state of Schleswig-Holstein. In addition to a neurological and cognitive Wechsler Intelligence Scale for Children, 4th edition assessment, self- and parent-reported psychological problems (Strengths and Difficulties Questionnaire), and health-related quality of life (KINDLR) were obtained and compared with a general population of 3,737 adolescents using data from a German Health Interview and Examination Survey for Children and Adolescents (KiGGS survey 2003-2006, Robert Koch Institute, Germany). Results Overall, 72 of the 90 surviving adolescents and their families (80.0%) participated in this study (mean age: 15.7 years; range: 14.2-17.2). A total of 22 adolescents (30.6%) did not have any neurosensory or cognitive limitations, whereas 31 adolescents (43.1%) were affected by multiple impairments. Parent-reported psychological problems were more common in the study cohort for the total difficulties score (11.3 points [standard deviation, SD: 5.9] vs. 7.7 points [SD: 5.0], p < 0.001) and for three of the five domains (emotional symptoms, hyperactivity-inattention, peer relationship problems) compared with the KiGGS reference population. In contrast, the rate of self-reported "abnormal" psychological problems was similar to the reference population. Parent-reported HRQOL was significantly lower in the study cohort for the total score (-3.7 points, p < 0.05) and for three of six subscales (emotional wellbeing, p < 0.05; self-esteem, p < 0.001, and wellbeing with regard to friends/friendship, p < 0.001). The adolescents themselves reported an HRQOL similar to that of the KiGGS reference population, though they rated their HRQOL higher for wellbeing with regard to families (p < 0.001) and wellbeing with regard to school/all-day function (p < 0.001). Overall, the parent- and self-reported HRQOL results were not associated with maternal education or disability status but were associated with adolescents' psychological problems. Conclusion Concordant with the high rate of functional and cognitive limitations, parents of adolescents who were born extremely prematurely faced more psychological problems in their children and reported a lower HRQOL compared with the KiGGS reference population. The adolescents themselves did not recognize these differences. However, having psychological problems was associated with lower self- and parent-reported HRQOL results.


Laetoli footprints preserve earliest direct evidence of human-like bipedal biomechanics.
Debates over the evolution of hominin bipedalism, a defining human characteristic, revolve around whether early bipeds walked more like humans, with energetically efficient extended hind limbs, or more like apes with flexed hind limbs. The 3.6 million year old hominin footprints at Laetoli, Tanzania represent the earliest direct evidence of hominin bipedalism. Determining the kinematics of Laetoli hominins will allow us to understand whether selection acted to decrease energy costs of bipedalism by 3.6 Ma. Using an experimental design, we show that the Laetoli hominins walked with weight transfer most similar to the economical extended limb bipedalism of humans. Humans walked through a sand trackway using both extended limb bipedalism, and more flexed limb bipedalism. Footprint morphology from extended limb trials matches weight distribution patterns found in the Laetoli footprints. These results provide us with the earliest direct evidence of kinematically human-like bipedalism currently known, and show that extended limb bipedalism evolved long before the appearance of the genus Homo. Since extended-limb bipedalism is more energetically economical than ape-like bipedalism, energy expenditure was likely an important selection pressure on hominin bipeds by 3.6 Ma.


Left ventricular size, output, and structure during guinea pig pregnancy.
We investigated left ventricular (LV) hemodynamics, pressure-volume relations, and morphometry to determine what cardiac changes characterize pregnancy in the guinea pig. Time-bred virgin guinea pigs were paired by weight with unbred controls. Hemodynamic studies and LV pressure-volume relations were obtained on days 59-68 of the 68-day gestation. Weight of control sows increased from 817 to 902 g (P less than 0.01) and pregnant sows from 810 to 1,251 g (P less than 0.01). LV weights were not different. When indexed for maternal weight minus uterus and contents, pregnancy produced increases in O2 consumption, +48% (P less than 0.01); cardiac output, +32% (P less than 0.05); and stroke volume, +46% (P less than 0.025). Passive LV pressure-volume curves (dP/dV) were shifted to the right (P less than 0.025), but dP/dV at constant pressure was unchanged. Using a thin-walled spherical model, elastic modulus at constant stress was not different. The percent LV inter- and intracellular volumes and myocyte myofibril and organelle volumes were unchanged during pregnancy. In the guinea pig, pregnancy increases LV output, stroke volume, and size without changes in LV mass, morphometry, or elastic modulus.


Sex determination from the ribs of contemporary Turks.
There have been several in-depth studies showing that the sternal extremity of the fourth rib can be used in estimating age as well as in determining sex, as illustrated in North American whites and blacks. Yet biological differences between populations may preclude the cross-application of standards. To test this hypothesis and develop a sex determination standard for the Turkish population, a sample of the sternal ends of the fourth rib was collected from 294 individuals autopsied in Istanbul, Turkey. Dimensions from the superior edge to the inferior one (SI) and the anterior edge to its posterior edge (AP) were measured with a caliper. The sample was divided into three age groups, "young," "old," and "total." Using discriminant function analysis, three formulae were developed, one for each group. The results indicated that SI height is the most dimorphic dimension and that both dimensions together can given an accuracy of 86% to 90%. To reach such an accuracy, the age of the unknown should be estimated, at least whether it is phase 4 or above or below, using the age standards for ribs developed by Işcan and associates. When cross-validation tests are carried out, incorrect formulae reduce accuracy by as much as 20%. Furthermore, it was observed that North American based white sex determination formulae assign many Turkish males into a female category. In conclusion, it is important to note that population and age specificity are essential in determining sex from the rib. Yet its accuracy is as good as, if not better than, many bones of the postcranial skeleton.


Ebp2 and Brx1 function cooperatively in 60S ribosomal subunit assembly in Saccharomyces cerevisiae.
The yeast protein Ebp2 is required for early steps in production of 60S ribosomal subunits. To search for cofactors with which Ebp2 functions, or substrates on which it acts, we screened for mutants that were synthetically lethal (sl) with the ebp2-14 mutation. Four different mutant alleles of the 60S ribosomal subunit assembly factor Brx1 were found. To investigate defects of the double mutant, we constructed strains conditional for the ebp2-14 brx1- synthetic lethal phenotype. These ebp2-14 brx1 mutants were defective in processing of 27S pre-rRNA and production of 60S subunits, under conditions where each single mutant was not. Ebp2 and Brx1 exhibit a strong two-hybrid interaction, which is eliminated by some combinations of brx1 and ebp2 mutations. In one such mutant, Ebp2 and Brx1 can still associate with pre-ribosomes, but subunit maturation is perturbed. Depletion of either Ebp2 or Brx1 revealed that Brx1 requires Ebp2 for its stable association with pre-ribosomes, but Ebp2 does not depend on the presence of Brx1 to enter pre-ribosomes. These results suggest that assembly of 60S ribosomal subunits requires cooperation of Ebp2 with Brx1, together with other molecules present in pre-ribosomes, potentially including several found in assembly subcomplexes with Brx1 and Ebp2.


Effectiveness of the diabetic foot risk classification system of the International Working Group on the Diabetic Foot.
To evaluate the effectiveness of a diabetic foot risk classification system by the International Working Group on the Diabetic Foot to predict clinical outcomes. A total of 225 diabetic patients were initially evaluated as part of a prospective case-control study at the University of Texas Health Science Center at San Antonio. Complete records were available for 213 patients for follow-up evaluation after 29 months. Upon enrollment, subjects were stratified into four risk groups based on the presence of risk factors according to the consensus of the International Working Group on the Diabetic Foot. Group 0 consisted of subjects without neuropathy, group 1 consisted of patients with neuropathy but without deformity or peripheral vascular disease (PVD), group 2 consisted of subjects with neuropathy and deformity or PVD, and group 3 consisted of patients with a history of foot ulceration or a lower-extremity amputation. Upon enrollment, patients in higher-risk groups had longer duration of diabetes, worse glycemic control, vascular and neuropathic variables, and more systemic complications of diabetes. During 3 years of follow-up, ulceration occurred in 5.1, 14.3, 18.8, and 55.8% of the patients in groups 0, 1, 2, and 3, respectively (linear-by-linear association, P < 0.001). All amputations were found in Groups 2 and 3 (3.1 and 20.9%, P < 0.001). The foot risk classification of the International Working Group on the Diabetic Foot predicts ulceration and amputation and can function as a tool to prevent lower-extremity complications of diabetes.


Caspase-8 Deficient Osteoblastic Cells Display Alterations in Non-Apoptotic Pathways.
Caspase-8 is the key component of the receptor-mediated (extrinsic) apoptotic pathway. Immunological localization of active caspase-8 showed its presence in osteoblasts, including non-apoptotic ones. Further in vivo exploration of caspase-8 functions in the bone is hindered by the fact that the caspase-8 knock-out is lethal prenatally. Examinations were thus performed using individual cell populations in vitro. In this study, caspase-8 was eliminated by the CRISPR/cas9 technology in MC3T3-E1 cells, the most common in vitro model of osteoblastic populations. The aim of the work was to specify the consequences of caspase-8 deficiency on non-apoptotic pathways. The impact on the osteogenic gene expression of the osteoblastic cells along with alterations in proliferation, caspase cascades and rapamycin induced autophagy response were evaluated. Osteogenic differentiation of caspase-8 deficient cells was inhibited as these cells displayed a decreased level of mineralization and lower activity of alkaline phosphatase. Among affected osteogenic genes, based on the PCR Array, major changes were observed for Ctsk, as down-regulated, and Gdf10, as up-regulated. Other significantly down-regulated genes included those coding osteocalcin, bone morphogenetic proteins (-3, -4 and -7), collagens (-1a1, -14a1) or Phex. The formation of autophagosomes was not altered in rapamycin-treated caspase-8 deficient cells, but expression of some autophagy-related genes, including Tnfsf10, Cxcr4, Dapk1 and Igf1, was significantly downregulated. These data provide new insight into the effects of caspase-8 on non-apoptotic osteogenic pathways.


A Plasma Biomarker Panel to Identify Surgically Resectable Early-Stage Pancreatic Cancer.
Blood-based biomarkers for early detection of pancreatic ductal adenocarcinoma (PDAC) are urgently needed. Current biomarkers lack high sensitivity and specificity for population screening. The gold-standard biomarker, CA 19-9, also fails to demonstrate the predictive value necessary for early detection. To validate a functional genomics-based plasma migration signature biomarker panel, plasma tissue factor pathway inhibitor (TFPI), tenascin C (TNC-FN III-C), and CA 19-9 levels were measured by enzyme-linked immunosorbent assays in three early-stage PDAC plasma cohorts, including two independent blinded validation cohorts containing a total of 43 stage I, 163 stage II, 86 chronic pancreatitis, 31 acute biliary obstruction, and 108 controls. Logistic regression models developed classification rules combining TFPI and/or TNC-FN III-C with CA 19-9 for patient cases and control subjects, with or without adjustment for age and diabetes status. Model classification performance was evaluated and analyses repeated among subpopulations without diabetes and pancreatitis history. Two-sided P values were calculated using bootstrap method. The TFPI/TNC-FN III-C/CA 19-9 panel improved CA 19-9 performance in all early-stage cohorts, including discriminating stage IA/IB/IIA, stage IIB, and all early-stage cancer from healthy controls. Statistical significance was reached for a number of subcohorts, including for all early-stage cancer vs healthy controls (cohort 1 AUC = 0.92, 95% CI = 0.86 to 0.96, P = .04; cohort 3 AUC = 0.83, 95% CI = 0.76 to 0.89, P = .045). Among subcohorts without diabetes and pancreatitis history, the panel approaches potential clinical utility for early detection to discriminate early-stage PDAC from healthy controls including an area under the curve (AUC) of 0.87 (95% CI = 0.77 to 0.95) for stage I/IIA, an AUC of 0.93 (95% CI = 0.87 to 0.98) for stage IIB, and a statistically significant AUC of 0.89 (95% CI = 0.82 to 0.95) for all early-stage cancer ( P = .03). TFPI/TNC-FN III-C migration signature adds statistically significantly to CA 19-9's predictive power to detect early-stage PDAC and may have clinical utility for early detection of surgically resectable PDAC, as well as for enhanced survival from this routinely lethal cancer.


Fruit growth in Arabidopsis occurs via DELLA-dependent and DELLA-independent gibberellin responses.
Fruit growth and development depend on highly coordinated hormonal activities. The phytohormone gibberellin (GA) promotes growth by inducing degradation of the growth-repressing DELLA proteins; however, the extent to which DELLA proteins contribute to GA-mediated gynoecium and fruit development remains to be clarified. Here, we provide an in-depth characterization of the role of DELLA proteins in Arabidopsis thaliana fruit growth. We show that DELLA proteins are key regulators of reproductive organ size and important for ensuring optimal fertilization. We demonstrate that the seedless fruit growth (parthenocarpy) observed in della mutants can be directly attributed to the constitutive activation of GA signaling. It has been known for >75 years that another hormone, auxin, can induce formation of seedless fruits. Using mutants with complete lack of DELLA activity, we show here that auxin-induced parthenocarpy occurs entirely through GA signaling in Arabidopsis. Finally, we uncover the existence of a DELLA-independent GA response that promotes fruit growth. This response requires GIBBERELLIN-INSENSITIVE DWARF1-mediated GA perception and a functional 26S proteasome and involves the basic helix-loop-helix protein SPATULA as a key component. Taken together, our results describe additional complexities in GA signaling during fruit development, which may be particularly important to optimize the conditions for successful reproduction.


Preclinical challenges for developing long acting intravitreal medicines.
The majority of blinding conditions arise due to chronic pathologies in the retina. During the last two decades, antibody-based medicines administered by intravitreal injection directly into the back of the eye have revolutionised the treatment of chronic retinal diseases characterised by uncontrolled blood vessel growth, e.g. wet age-related macular degeneration (wAMD), diabetic retinopathy (DR) and choroidal neovascularisation (CNV). Although intravitreal injections have become a commonly performed ophthalmic procedure that provides a reproducible dose to maximise drug exposure in the back of the eye, there is a need to minimise the frequency and cumulative number of intravitreal injections. Developing longer acting intraocular therapies is one key strategy that is being pursued. Pharmaceutical preclinical development of intraocular medicines is heavily reliant on the use of animal models to determine ocular tolerability, pharmacokinetics, biodistribution and drug stability. Animal eyes are different from human eyes, such as the anatomy, organisation of vitreous macromolecular structure, aqueous outflow and immune response; all which impacts the ability to translate preclinical data into a clinical product. The development of longer acting protein formulations using animals is also limited because animals reject human proteins. Preclinical strategies also do not account for differences in the vitreous due to ageing and whether a vitrectomy has been performed. Intraocular formulations must reside and clear from the vitreous body, so there is a need for the formulation scientist to have knowledge about vitreous structure and physiology to facilitate preclinical development strategies. Preclinical pharmaceutical development paradigms used to create therapies for other routes of administration (e.g. oral, subcutaneous, pulmonary and intravenous) are grounded on the use of preclinical in vitro models. Analogous pharmaceutical strategies with appropriately designed in vitro models that can account for intraocular mass transfer to estimate pharmacokinetic profiles can be used to develop in vitro-in vivo correlations (IVIVCs) to accelerate the preclinical optimisation of long-acting intraocular formulations. Data obtained can then inform preclinical in vivo and clinical studies. With the now widespread use of intravitreal injections, it is also important during early preclinical studies to ensure there is a viable regulatory pathway for new therapies. Knowledge of the physiological, pharmaceutical and regulatory factors will help in the development of long-acting intravitreal medicines, which is rapidly evolving into a distinct pharmaceutical discipline.


Assessing the anti-tumour properties of Iraqi propolis in vitro and in vivo.
The study was designed to evaluate anti-tumour properties of Iraqi propolis collected from Mosul region (M) on HL-60 and HCT-116 cell lines and on HCT-116 in vivo. M induced an inhibitory effect against the proliferation of HL-60 and colony potential of HCT-116 cells. The apoptosis in HL-60 cells was associated with down-regulation of Bcl-2 and activation of Bax, while in HCT-116 cells, necrotic features were observed; size of cells was dramatically increased by swelling of cytoplasm and loss of membrane integrity, cell rupture and release of cellular contents. Analysis of BrdU/DNA cell cycle in both cell lines showed that M induced cell cycle perturbations in both BrdU positive and BrdU negative cells. The exposure of HL-60 to M caused γ-H2AX in a dose dependent manner and was associated with induction of apoptosis. The experiments in HCT-116 tumor-bearing mice showed that oral administration of propolis at doses that caused no detectable toxicity was associated with a decrease in mitotic cells and an increase in endoreduplications, increased p53 and decreased Ki-67 expression of cells in tumor sections. This study provides the rationale to investigate the potential beneficial effect of propolis in the diet of patients receiving anti-cancer therapies.


Computational study of scattering from healthy and diseased red blood cells.
We present a comparative study of scattering from healthy red blood cells (RBCs) and diseased RBCs with deformed shapes. Scattering problems involving three-dimensional RBCs are formulated accurately with the electric and magnetic current combined-field integral equation and solved efficiently by the multilevel fast multipole algorithm. We compare scattering cross section values obtained for different RBC shapes and different orientations. In this way, we determine strict guidelines to distinguish deformed RBCs from healthy RBCs and to diagnose various diseases using scattering cross section values. The results may be useful for designing new and improved flow cytometry procedures.


Multiple layers of chimerism in a single-stranded DNA virus discovered by deep sequencing.
Viruses with single-stranded (ss) DNA genomes infect hosts in all three domains of life and include many medically, ecologically, and economically important pathogens. Recently, a new group of ssDNA viruses with chimeric genomes has been discovered through viral metagenomics. These chimeric viruses combine capsid protein genes and replicative protein genes that, respectively, appear to have been inherited from viruses with positive-strand RNA genomes, such as tombusviruses, and ssDNA genomes, such as circoviruses, nanoviruses or geminiviruses. Here, we describe the genome sequence of a new representative of this virus group and reveal an additional layer of chimerism among ssDNA viruses. We show that not only do these viruses encompass genes for capsid proteins and replicative proteins that have distinct evolutionary histories, but also the replicative genes themselves are chimeras of functional domains inherited from viruses of different families. Our results underscore the importance of horizontal gene transfer in the evolution of ssDNA viruses and the role of genetic recombination in the emergence of novel virus groups.


Risk of Reactivation of Hepatitis B Virus (HBV) and Tuberculosis (TB) and Complications of Hepatitis C Virus (HCV) Following Tocilizumab Therapy: A Systematic Review to Inform Risk Assessment in the COVID-19 Era.
Objectives: Tocilizumab (TCZ), an IL-6 receptor antagonist, is used in the treatment of severe COVID-19 caused by infection with SARS-CoV-2. However, unintended consequences of TCZ therapy include reactivation of tuberculosis (TB) or hepatitis B virus (HBV), and worsening of hepatitis C virus (HCV). We set out to assimilate existing data for these complications, in order to help inform evidence-based risk assessments for the use of TCZ, and thus to reduce the risk of serious but preventable complications. Methods: We searched the global WHO database of Individual Case Safety Reports (ICSRs) and adverse drug reactions (ADRs) ("VigiBase") and undertook a systematic literature review, in accordance with PRISMA guidelines. We generated mean cumulative incidence estimates for infection complications. Results: Mean cumulative incidence of HBV and TB were 3.3 and 4.3%, respectively, in patients receiving TCZ. Insufficient data were available to generate estimates for HCV. These estimates derive from heterogeneous studies pre-dating SARS-CoV-2, with differing epidemiology and varied approaches to screening and prophylaxis, so formal meta-analysis was not possible. Conclusions: We underline the need for careful individual risk assessment prior to TCZ prescription, and present an algorithm to guide clinical stratification. There is an urgent need for ongoing collation of safety data as TCZ therapy is used in COVID.


The prevalence of confirmed maltreatment among US children, 2004 to 2011.
Child maltreatment is a risk factor for poor health throughout the life course. Existing estimates of the proportion of the US population maltreated during childhood are based on retrospective self-reports. Records of officially confirmed maltreatment have been used to produce annual rather than cumulative counts of maltreated individuals. To estimate the proportion of US children with a report of maltreatment (abuse or neglect) that was indicated or substantiated by Child Protective Services (referred to as confirmed maltreatment) by 18 years of age. The National Child Abuse and Neglect Data System (NCANDS) Child File includes information on all US children with a confirmed report of maltreatment, totaling 5,689,900 children (2004-2011). We developed synthetic cohort life tables to estimate the cumulative prevalence of confirmed childhood maltreatment by 18 years of age. The cumulative prevalence of confirmed child maltreatment by race/ethnicity, sex, and year. At 2011 rates, 12.5% (95% CI, 12.5%-12.6%) of US children will experience a confirmed case of maltreatment by 18 years of age. Girls have a higher cumulative prevalence (13.0% [95% CI, 12.9%-13.0%]) than boys (12.0% [12.0%-12.1%]). Black (20.9% [95% CI, 20.8%-21.1%]), Native American (14.5% [14.2%-14.9%]), and Hispanic (13.0% [12.9%-13.1%]) children have higher prevalences than white (10.7% [10.6%-10.8%]) or Asian/Pacific Islander (3.8% [3.7%-3.8%]) children. The risk for maltreatment is highest in the first few years of life; 2.1% (95% CI, 2.1%-2.1%) of children have confirmed maltreatment by 1 year of age, and 5.8% (5.8%-5.9%), by 5 years of age. Estimates from 2011 were consistent with those from 2004 through 2010. Annual rates of confirmed child maltreatment dramatically understate the cumulative number of children confirmed to be maltreated during childhood. Our findings indicate that maltreatment will be confirmed for 1 in 8 US children by 18 years of age, far greater than the 1 in 100 children whose maltreatment is confirmed annually. For black children, the cumulative prevalence is 1 in 5; for Native American children, 1 in 7.


Chemical Modifications Mark Alternatively Spliced and Uncapped Messenger RNAs in Arabidopsis.
Posttranscriptional chemical modification of RNA bases is a widespread and physiologically relevant regulator of RNA maturation, stability, and function. While modifications are best characterized in short, noncoding RNAs such as tRNAs, growing evidence indicates that mRNAs and long noncoding RNAs (lncRNAs) are likewise modified. Here, we apply our high-throughput annotation of modified ribonucleotides (HAMR) pipeline to identify and classify modifications that affect Watson-Crick base pairing at three different levels of the Arabidopsis thaliana transcriptome (polyadenylated, small, and degrading RNAs). We find this type of modifications primarily within uncapped, degrading mRNAs and lncRNAs, suggesting they are the cause or consequence of RNA turnover. Additionally, modifications within stable mRNAs tend to occur in alternatively spliced introns, suggesting they regulate splicing. Furthermore, these modifications target mRNAs with coherent functions, including stress responses. Thus, our comprehensive analysis across multiple RNA classes yields insights into the functions of covalent RNA modifications in plant transcriptomes.


Continual development of a personalized decision support system.
Improving the safety, quality, and efficiency of care with the help of clinical decision support tools is one of the core objectives in the meaningful use of Electronic Health Records. Successful adoption of support systems depends on the quality of delivered information, its relevance to the clinical task and individual patient, integration of the system with the entire clinical workplace, and ease of use of the system. This paper presents continuous development and evaluation, as well as lessons learned in development and maintenance of an evidence-based system that supports development of individualized patient care plans. Since its deployment in August 2009, the Evidence-Based Practice InfoBot (EBP InfoBot) system is in daily use at the NIH Clinical Center and responds to 21 requests a day, on average.


Tight packaging of erbium-doped waveguide amplifiers.
A numerical model for erbium-doped waveguide amplifiers (EDWAs) containing bent waveguides was developed. The model uses rate-propagation equations and takes into account bend-induced losses in the waveguide with a varying radius of curvature. It is applied to the design of tightly packed erbium-doped waveguide amplifiers under two constraints: minimization of the area required to obtain a predetermined gain and maximization of the gain available from a given area. Numerical calculations based on realistic waveguide parameters demonstrate the possibility of fabricating high-gain, small-size EDWAs. Simple design rules, based on geometric formulas and consideration of bend losses, are given and compared with the results of numerical calculations.