Treatment of chemotherapy-induced leukopenia in a rat model with aqueous extract from Uncaria tomentosa.
The Uncaria tomentosa water extracts (C-Med-100) depleted of indole alkaloids (< 0.05%, w/w) have been shown to induce apoptosis and inhibit proliferation in tumor cells in vitro and to enhance DNA repair, mitogenic response and white blood cells in vivo. In this study, the effect of C-Med-100 in the treatment of chemically induced leukopenia was evaluated in a rat model. W/Fu rats were treated first with doxorubicin (DXR) 2 mg/kg x 3 (i.p. injection at 24 hour-intervals) to induce leukopenia. Twenty-four hours after the last DXR treatment, the rats were daily gavaged with C-Med-100 for 16 consecutive days. As a positive control, Neupogen, a granulocyte colony stimulator was also administered by subcutaneous injection at a dose of 5 and 10 microg/ml for 10 consecutive days. The results showed that both C-Med-100 and Neupogen treatment groups recovered significantly sooner (p < 0.05 by Duncan test) than DXR group. However, the recovery by C-Med-100 treatment was a more natural process than Neupogen because all fractions of white blood cells were proportionally increased while Neupogen mainly elevated the neutrophil cells. These results were also confirmed by microscopic examination of the blood smears. The mechanism of the C-Med-100 effect on WBC is not known but other data showing enhanced effects on DNA repair and immune cell proliferative response support a general immune enhancement.


Growth Differentiation Factor 15 and NT-proBNP as Blood-Based Markers of Vascular Brain Injury and Dementia.
Background GDF15 (growth differentiation factor 15) and NT-proBNP (N-terminal pro-B-type natriuretic peptide) may offer promise as biomarkers for cognitive outcomes, including dementia. We determined the association of these biomarkers with cognitive outcomes in a community-based cohort. Methods and Results Plasma GDF15 (n=1603) and NT-proBNP levels (n=1590) (53% women; mean age, 68.7 years) were measured in dementia-free Framingham Offspring cohort participants at examination 7 (1998-2001). Participants were followed up for incident dementia. Secondary outcomes included Alzheimer disease dementia, magnetic resonance imaging structural brain measures, and neurocognitive performance. During a median 11.8-year follow-up, 131 participants developed dementia. On multivariable Cox proportional-hazards analysis, higher circulating GDF15 was associated with an increased risk of incident all-cause and Alzheimer disease dementia (hazard ratio [HR] per SD increment in natural log-transformed biomarker value, 1.54 [95% CI, 1.22-1.95] and 1.37 [95% CI, 1.03-1.81], respectively), whereas higher plasma NT-proBNP was also associated with an increased risk of all-cause dementia (HR, 1.32; 95% CI, 1.05-1.65). Elevated GDF15 was associated with lower total brain and hippocampal volumes, greater white matter hyperintensity volume, and poorer cognitive performance. Elevated NT-proBNP was associated with greater white matter hyperintensity volume and poorer cognitive performance. Addition of both biomarkers to a conventional risk factor model improved dementia risk classification (net reclassification improvement index, 0.25; 95% CI, 0.05-0.45). Conclusions Elevated plasma GDF15 and NT-proBNP were associated with vascular brain injury on magnetic resonance imaging, poorer neurocognitive performance, and increased risk of incident dementia in individuals aged >60 years. Both biomarkers improved dementia risk classification beyond that of traditional clinical risk factors, indicating their potential value in predicting incident dementia.


Cloning of human lymphocyte-specific interferon regulatory factor (hLSIRF/hIRF4) and mapping of the gene to 6p23-p25.
The interferon regulatory factor (IRF) genes encode a family of transcription factors involved in the transcriptional regulation of interferon and the interferon stimulated genes through recognition of the interferon stimulated response element. We previously reported the cloning of a murine lymphocyte-specific IRF (mLSIRF), which was rapidly induced following B- or T-cell receptor crosslinking. To study the role of LSIRF in human lymphocyte development, we have cloned the complete 5.3-kb cDNA for the human homolog (hLSIRF). hLSIRF is a protein of 450 amino acids with a predicted molecular weight of 51.6 kDa and possesses 92% identity at the amino acid level to mLSIRF, including near identity in the DNA-binding domain. In Northern blot analysis, a single transcript of approximately 5 kb was highly expressed in spleen and peripheral blood lymphocyte. hLSIRF mRNA was rapidly induced in peripheral T cells after crosslinking the T-cell receptor. Analysis of tumor cell lines showed that hLSIRF mRNA was basally expressed in most B- but not T-cell lines. Surprisingly hLSIRF mRNA was also found in the melanoma line G361 and is expressed in normal melanocytes as well. Sequence from a genomic clone for hLSIRF was compared to that from mouse and revealed an identical exon-intron structure and a conserved PU-1-binding motif in the promoter. By FISH analysis, hLSIRF was mapped to 6p23-p25.


The Epstein-Barr virus encoded LMP1 oncoprotein modulates cell adhesion via regulation of activin A/TGFβ and β1 integrin signalling.
Approximately 20% of global cancer incidence is causally linked to an infectious agent. Epstein-Barr virus (EBV) accounts for around 1% of all virus-associated cancers and is associated with nasopharyngeal carcinoma (NPC). Latent membrane protein 1 (LMP1), the major oncoprotein encoded by EBV, behaves as a constitutively active tumour necrosis factor (TNF) receptor activating a variety of signalling pathways, including the three classic MAPKs (ERK-MAPK, p38 MAPK and JNK/SAPK). The present study identifies novel signalling properties for this integral membrane protein via the induction and secretion of activin A and TGFβ1, which are both required for LMP1's ability to induce the expression of the extracellular matrix protein, fibronectin. However, it is evident that LMP1 is unable to activate the classic Smad-dependent TGFβ signalling pathway, but rather elicits its effects through the non-Smad arm of TGFβ signalling. In addition, there is a requirement for JNK/SAPK signalling in LMP1-mediated fibronectin induction. LMP1 also induces the expression and activation of the major fibronectin receptor, α5β1 integrin, an effect that is accompanied by increased focal adhesion formation and turnover. Taken together, these findings support the putative role for LMP1 in the pathogenesis of NPC by contributing to the metastatic potential of epithelial cells.


To reduce or abstain? Substance use goals in the treatment of veterans with substance use disorders and comorbid PTSD.
Posttraumatic stress disorder (PTSD) and substance use disorders (SUD) frequently co-occur. Previous research demonstrates the utility of goals in attaining improved SUD outcomes, however, no previous studies have examined goal choices in the context of integrated treatment for comorbid PTSD and SUD. The present study investigated correlates of treatment entry goals to either reduce or abstain from substance use. Participants (N = 60) were treatment-seeking veterans with current PTSD and SUD. Participants completed self-report and clinician-rated measures of substance use, PTSD, and affective symptoms as part of a larger randomized controlled trial. Half (30/60) of participants endorsed a treatment entry goal to reduce substance use (reducers). Compared to participants who endorsed a treatment entry goal of abstinence (abstainers), reducers were significantly younger, more likely to be employed, more likely to have served in recent military conflicts (Operations Enduring/Iraqi Freedom), and endorsed significantly fewer symptoms of alcohol dependence. The findings demonstrate clinically relevant differences based on treatment entry goals, suggesting that individuals are often able to choose conceivably appropriate treatment goals based, most notably, on the severity of their SUD. Collaboratively engaging patients in establishing treatment goals that are consistent with their beliefs and desires in conjunction with empirical findings is particularly relevant in the context of treatment for SUD and PTSD where many patients are ambivalent about treatment and attrition is common.


A multidisciplinary, comprehensive, ambulatory treatment scheme for diabetes mellitus in children.
A study has been carried out on 262 children with juvenile diabetes and their parents, treated up to 10 yr on an ambulatory basis by a multidisciplinary team composed of pediatric endocrinologist, nurse, dietitian, psychologist, and social worker. Comparison of the findings with those of a study performed before inception of the Counselling Center for Juvenile Diabetics revealed the following positive influences: the degree of control attained was both higher and sustained with greater regularity; there were fewer complications with no episodes of coma, brittle diabetes, or severe ketoacidosis and almost no need for hospitalization; the attitude of the affected child, his parents, and his teachers was found to be considerably improved; there was better understanding of the nature of the disease and its requirements; the child's motivation to maintain the diabetic regimen was greater and conflicts within the family circle were markedly reduced; the child's self-concept was much higher; and both scholastic achievements and social adjustment were greater. We concluded that psychological stability is a basic factor in the control of diabetes, and the value of the multidisciplinary approach in the treatment of this chronic disease is indicated.


Classification of Metastatic versus Non-Metastatic Axillary Nodes in Breast Cancer Patients: Value of Cortex-Hilum Area Ratio with Ultrasound.
To investigate the significance of the cortex-hilum (CH) area ratio and longitudinal-transverse (LT) axis ratio and the blood flow pattern for diagnosis of metastatic axillary lymph nodes by ultrasound in breast cancer patients. From October 2005 to July 2006, we prospectively evaluated axillary nodes with ultrasound in 205 consecutive patients who had category 4B, 4C or 5 breast lesions according to the Breast Imaging Reporting and Data System-Ultrasound (BI-RADS-Ultrasound®). Among the 205, there were 24 patients who had pathologic verification of breast cancer and axillary lymph node status. For a total of 80 axillary nodes we measured the areas of the cortex and hilum of lymph nodes and calculated the area ratio. We also measured the length of the longitudinal and transverse axis of the lymph nodes and calculated the length ratio. We evaluated the blood flow pattern on power Doppler imaging and classified each lymph node into a central or peripheral pattern. Diagnostic performance was analyzed according to positive criteria for lymph node metastasis (CH area ratio >2, LT axis ratio <2, peripheral type on power Doppler imaging). The sensitivity of the CH area ratio was superior to that of the LT axis ratio (94.1% vs. 82.3%, p=0.031) and to that of the blood flow pattern (94.1% vs. 29.4%, p=0.009). For specificity, all three evaluating parameters had high values (89.1-95.6%) and no significant differences were found (p=0.121). The CH area ratio had a better positive predictive value than the LT axis ratio (94.1% vs. 80.0%, p=0.030) and power Doppler imaging (94.1% vs. 66.6%, p=0.028). For the negative predictive value, the CH area ratio was superior to the LT axis ratio (95.6% vs. 86.6%, p=0.035) and the blood flow pattern (95.6% vs. 63.0%, p=0.027). We recommend the CH area ratio of an axillary lymph node on ultrasound as a quantitative indicator for the classification of lymph nodes. The CH area ratio can improve diagnostic performance when compared with the LT axis ratio or blood flow pattern.


Evaluation of changes in the tumor microenvironment after sorafenib therapy by sequential histology and 18F-fluoromisonidazole hypoxia imaging in renal cell carcinoma.
The mechanistic dissociation of 'tumor starvation' versus 'vascular normalization' following anti-angiogenic therapy is a subject of intense controversy in the field of experimental research. In addition, accurately evaluating changes of the tumor microenvironment after anti-angiogenic therapy is important for optimizing treatment strategy. Sorafenib has considerable anti-angiogenic effects that lead to tumor starvation and induce tumor hypoxia in the highly vascularized renal cell carcinoma (RCC) xenografts. 18F-fluoromisonidazole (18F‑FMISO) is a proven hypoxia imaging probe. Thus, to clarify early changes in the tumor microenvironment following anti-angiogenic therapy and whether 18F-FMISO imaging can detect those changes, we evaluated early changes in the tumor microenvironment after sorafenib treatment in an RCC xenograft by sequential histological analysis and 18F-FMISO autoradiography (ARG). A human RCC xenograft (A498) was established in nude mice, for histological studies and ARG, and further assigned to the control and sorafenib-treated groups (80 mg/kg, per os). Mice were sacrificed on Days 1, 2, 3 and 7 in the histological study, and on Days 3 and 7 in ARG after sorafenib treatment. Tumor volume was measured every day. 18F-FMISO and pimonidazole were injected intravenously 4 and 2 h before sacrifice, respectively. Tumor sections were stained with hematoxylin and eosin and immunohistochemically with pimonidazole and CD31. Intratumoral 18F-FMISO distribution was quantified in ARG. Tumor volume did not significantly change on Day 7 after sorafenib treatment. In the histological study, hypoxic fraction significantly increased on Day 2, mean vessel density significantly decreased on Day 1 and necrosis area significantly increased on Day 2 after sorafenib treatment. Intratumoral 18F-FMISO distribution significantly increased on Days 3 (10.2-fold, p<0.01) and 7 (4.1-fold, p<0.01) after sorafenib treatment. The sequential histological evaluation of the tumor microenvironment clarified tumor starvation in A498 xenografts treated with sorafenib. 18F-FMISO hypoxia imaging confirmed the tumor starvation. 18F-FMISO PET may contribute to determine an optimum treatment protocol after anti-angiogenic therapy.


Co-oxidation of 2-bromohydroquinone by renal prostaglandin synthase. Modulation of prostaglandin synthesis by 2-bromohydroquinone and glutathione.
Homogenates from rat renal papillae, a rich source of the prostaglandin (PG) H synthase system (PHS), metabolized [14C]2-bromohydroquinone, in the presence of arachidonic acid, to products which are covalently bound to protein. The co-oxidation of 2-bromohydroquinone caused a concentration-dependent stimulation in 6-keto-PGF1 alpha, thromboxane B2, PGF2 alpha, PGE2, and PGD2 formation. Glutathione (1 mM) caused a decrease in prostaglandin formation and inhibited the arachidonic acid-supported covalent binding of [14C]2-bromohydroquinone with the concomitant formation of [14C]2-bromohydroquinone-glutathione conjugates, oxidized glutathione, and an increase in the recovery of [14C]2-bromohydroquinone. NADPH also inhibited [14C]2-bromohydroquinone covalent binding, probably by reduction of the semiquinone radical back to the hydroquinone. Indomethacin and aspirin, inhibitors of the cyclooxygenase component of PHS, and propylthiouracil and methimazole, inhibitors of the hydroperoxidase component of PHS, inhibited the arachidonic acid-supported covalent binding of [14C]2-bromohydroquinone by 94%, 52%, 78%, and 79% respectively. These data suggest that 1) renal PHS may play a role in activating the nephrotoxin, 2-bromohydroquinone, and that 2) xenobiotic metabolism and its subsequent effects on glutathione levels can modulate renal prostaglandin synthesis.


Incidental findings in children with blunt head trauma evaluated with cranial CT scans.
Cranial computed tomography (CT) scans are frequently obtained in the evaluation of blunt head trauma in children. These scans may detect unexpected incidental findings. The objectives of this study were to determine the prevalence and significance of incidental findings on cranial CT scans in children evaluated for blunt head trauma. This was a secondary analysis of a multicenter study of pediatric blunt head trauma. Patients <18 years of age with blunt head trauma were eligible, with those undergoing cranial CT scan included in this substudy. Patients with coagulopathies, ventricular shunts, known previous brain surgery or abnormalities were excluded. We abstracted radiology reports for nontraumatic findings. We reviewed and categorized findings by their clinical urgency. Of the 43,904 head-injured children enrolled in the parent study, 15,831 underwent CT scans, and these latter patients serve as the study cohort. On 670 of these scans, nontraumatic findings were identified, with 16 excluded due to previously known abnormalities or surgeries. The remaining 654 represent a 4% prevalence of incidental findings. Of these, 195 (30%), representing 1% of the overall sample, warranted immediate intervention or outpatient follow-up. A small but important number of children evaluated with CT scans after blunt head trauma had incidental findings. Physicians who order cranial CTs must be prepared to interpret incidental findings, communicate with families, and ensure appropriate follow-up. There are ethical implications and potential health impacts of informing patients about incidental findings.


Yeast RAD26, a homolog of the human CSB gene, functions independently of nucleotide excision repair and base excision repair in promoting transcription through damaged bases.
RAD26 in the yeast Saccharomyces cerevisiae is the counterpart of the human Cockayne syndrome group B (CSB) gene. Both RAD26 and CSB act in the preferential repair of UV lesions on the transcribed strand, and in this process, they function together with the components of nucleotide excision repair (NER). Here, we examine the role of RAD26 in the repair of DNA lesions induced upon treatment with the alkylating agent methyl methanesulfonate (MMS). MMS-induced DNA lesions include base damages such as 3-methyl adenine and 7-methyl guanine, and these lesions are removed in yeast by the alternate competing pathways of base excision repair (BER), which is initiated by the action of MAG1-encoded N-methyl purine DNA glycosylase, and NER. Interestingly, a synergistic increase in MMS sensitivity was observed in the rad26 Delta strain upon inactivation of NER or BER, indicating that RAD26 promotes the survival of MMS-treated cells by a mechanism that acts independently of either of these repair pathways. The galactose-inducible transcription of the GAL2, GAL7, and GAL10 genes is reduced in MMS-treated rad26 Delta cells and also in mag1 Delta rad14 Delta cells, whereas a very severe reduction in transcription occurs in MMS-treated mag1 Delta rad14 Delta rad26 Delta cells. From these observations, we infer that RAD26 plays a role in promoting transcription by RNA polymerase II through damaged bases. The implications of these observations are discussed in this paper.


Structural requirements for substrate recognition of Mycobacterium tuberculosis 14 alpha-demethylase: implications for sterol biosynthesis.
Sterol 14 alpha-demethylase (14DM) is a cytochrome P-450 involved in sterol biosynthesis in eukaryotes. It was reported that Mycobacterium smegmatis also makes cholesterol and that cholesterol is essential to Mycobacterium tuberculosis (MT) infection, although the origin of the cholesterol is unknown. A protein product from MT having about 30% sequence identity with eukaryotic 14 alpha-demethylases has been found to convert sterols to their 14-demethyl products indicating that a sterol pathway might exist in MT. To determine the optimal sterol structure recognized by MT 14DM, binding of 28 sterol and sterol-like (triterpenoids) molecules to the purified recombinant 14 alpha-demethylase was examined. Like eukaryotic forms, a 3 beta-hydroxy group and a 14 alpha-methyl group are essential for substrate acceptability by the bacterial 14 alpha-demethylase. The high affinity binding of 31-norcycloartenol without detectable activity indicates that the Delta(8)-bond is required for activity but not for binding. As for plant 14 alpha-demethylases, 31-nor-sterols show a binding preference for MT 14DM. Similar to enzymes from mammals and yeast, a C24-alkyl group is not required for MT 14DM binding and activity, whereas it is for plant 14 alpha-demethylases.Thus, substrate binding to MT 14DM seems to share common features with all eukaryotic 14 alpha-demethylases, the MT form seemingly having the broadest substrate recognition of all forms of 14 alpha-demethylase studied so far. - Bellamine, A., A. T. Mangla, A. L. Dennis, W. D. Nes, and M. R. Waterman. Structural requirements for substrate recognition of Mycobacterium tuberculosis 14 alpha-demethylase: implications for sterol biosynthesis. J. Lipid Res. 2001. 42: 128;-136.


Lower-limb joint mechanics during maximum acceleration sprinting.
We explored how humans adjust the stance phase mechanical function of their major lower-limb joints (hip, knee, ankle) during maximum acceleration sprinting. Experimental data [motion capture and ground reaction force (GRF)] were recorded from eight participants as they performed overground sprinting trials. Six alternative starting locations were used to obtain a dataset that incorporated the majority of the acceleration phase. Experimental data were combined with an inverse-dynamics-based analysis to calculate lower-limb joint mechanical variables. As forward acceleration magnitude decreased, the vertical GRF impulse remained nearly unchanged whereas the net horizontal GRF impulse became smaller as a result of less propulsion and more braking. Mechanical function was adjusted at all three joints, although more dramatic changes were observed at the hip and ankle. The impulse from the ankle plantar-flexor moment was almost always larger than those from the hip and knee extensor moments. Forward acceleration magnitude was linearly related to the impulses from the hip extensor moment (R2=0.45) and the ankle plantar-flexor moment (R2=0.47). Forward acceleration magnitude was also linearly related to the net work done at all three joints, with the ankle displaying the strongest relationship (R2=0.64). The ankle produced the largest amount of positive work (1.55±0.17 J kg-1) of all the joints, and provided a significantly greater proportion of the summed amount of lower-limb positive work as running speed increased and forward acceleration magnitude decreased. We conclude that the hip and especially the ankle represent key sources of positive work during the stance phase of maximum acceleration sprinting.


Defective postsecretory maturation of MUC5B mucin in cystic fibrosis airways.
In cystic fibrosis (CF), airway mucus becomes thick and viscous, and its clearance from the airways is impaired. The gel-forming mucins undergo an ordered "unpacking/maturation" process after granular release that requires an optimum postsecretory environment, including hydration and pH. We hypothesized that this unpacking process is compromised in the CF lung due to abnormal transepithelial fluid transport that reduces airway surface hydration and alters ionic composition. Using human tracheobronchial epithelial cells derived from non-CF and CF donors and mucus samples from human subjects and domestic pigs, we investigated the process of postsecretory mucin unfolding/maturation, how these processes are defective in CF airways, and the probable mechanism underlying defective unfolding. First, we found that mucins released into a normal lung environment transform from a compact granular form to a linear form. Second, we demonstrated that this maturation process is defective in the CF airway environment. Finally, we demonstrated that independent of HCO3- and pH levels, airway surface dehydration was the major determinant of this abnormal unfolding process. This defective unfolding/maturation process after granular release suggests that the CF extracellular environment is ion/water depleted and likely contributes to abnormal mucus properties in CF airways prior to infection and inflammation.


Ion channel density regulates switches between regular and fast spiking in soma but not in axons.
The threshold firing frequency of a neuron is a characterizing feature of its dynamical behaviour, in turn determining its role in the oscillatory activity of the brain. Two main types of dynamics have been identified in brain neurons. Type 1 dynamics (regular spiking) shows a continuous relationship between frequency and stimulation current (f-I(stim)) and, thus, an arbitrarily low frequency at threshold current; Type 2 (fast spiking) shows a discontinuous f-I(stim) relationship and a minimum threshold frequency. In a previous study of a hippocampal neuron model, we demonstrated that its dynamics could be of both Type 1 and Type 2, depending on ion channel density. In the present study we analyse the effect of varying channel density on threshold firing frequency on two well-studied axon membranes, namely the frog myelinated axon and the squid giant axon. Moreover, we analyse the hippocampal neuron model in more detail. The models are all based on voltage-clamp studies, thus comprising experimentally measurable parameters. The choice of analysing effects of channel density modifications is due to their physiological and pharmacological relevance. We show, using bifurcation analysis, that both axon models display exclusively Type 2 dynamics, independently of ion channel density. Nevertheless, both models have a region in the channel-density plane characterized by an N-shaped steady-state current-voltage relationship (a prerequisite for Type 1 dynamics and associated with this type of dynamics in the hippocampal model). In summary, our results suggest that the hippocampal soma and the two axon membranes represent two distinct kinds of membranes; membranes with a channel-density dependent switching between Type 1 and 2 dynamics, and membranes with a channel-density independent dynamics. The difference between the two membrane types suggests functional differences, compatible with a more flexible role of the soma membrane than that of the axon membrane.


Triangulating the sexually dimorphic brain through high-resolution neuroimaging of murine sex chromosome aneuploidies.
Murine sex chromosome aneuploidies (SCAs) provide powerful models for charting sex chromosome influences on mammalian brain development. Here, building on prior work in X-monosomic (XO) mice, we use spatially non-biased high-resolution imaging to compare and contrast neuroanatomical alterations in XXY and XO mice relative to their wild-type XX and XY littermates. First, we show that carriage of a supernumerary X chromosome in XXY males (1) does not prevent normative volumetric masculinization of the bed nucleus of the stria terminalis (BNST) and medial amygdala, but (2) causes distributed anatomical alterations relative to XY males, which show a statistically unexpected tendency to be co-localized with and reciprocal to XO-XX differences in anatomy. These overlaps identify the lateral septum, BNST, ventral group thalamic nuclei and periaqueductal gray matter as regions with replicable sensitivity to X chromosome dose across two SCAs. We then harness anatomical variation across all four karyotype groups in our study--XO, XX, XY and XXY--to create an agnostic data-driven segmentation of the mouse brain into five distributed clusters which (1) recover fundamental properties of brain organization with high spatial precision, (2) define two previously uncharacterized systems of relative volume excess in females vs. males ("forebrain cholinergic" and "cerebelo-pontine-thalamo-cortical"), and (3) adopt stereotyped spatial motifs which delineate ordered gradients of sex chromosome and gonadal influences on volumetric brain development. Taken together, these data provide a new framework for the study of sexually dimorphic influences on brain development in health and disrupted brain development in SCA.


Green tea polyphenols inhibit human vascular smooth muscle cell proliferation stimulated by native low-density lipoprotein.
This study investigated whether human vascular smooth muscle cell proliferation induced by native low-density lipoprotein (LDL) is affected by green tea catechins. Furthermore, the effects of native LDL on extracellular signal-regulated kinase (ERK) 1/2 activity were determined. Cell proliferation stimulated by native LDL was concentration-dependently inhibited by epigallocatechin, epigallocatechin-3-gallate, green tea polyphenon, and the nonspecific antioxidant N-acetylcysteine (P<0.05). Combined treatment of green tea polyphenon and N-acetylcysteine markedly potentiated the effect of each drug on vascular smooth muscle cell proliferation. ERK1/2 activity was only partly inhibited by green tea catechins alone or in combination with N-acetylcysteine (P<0.05). These data suggest that green tea constituents inhibit proliferation of human vascular smooth muscle cells exposed to high levels of native LDL. Green tea constituents and antioxidants may exert vascular protection by inhibiting human vascular smooth muscle cell growth associated with hypercholesterolemia.


Acute aerobic exercise differentially alters acylated ghrelin and perceived fullness in normal-weight and obese individuals.
Adiposity alters acylated ghrelin concentrations, but it is unknown whether adiposity alters the effect of exercise and feeding on acylated ghrelin responses. Therefore, the purpose of this study was to determine whether adiposity [normal-weight (NW) vs. obese (Ob)] influences the effect of exercise and feeding on acylated ghrelin, hunger, and fullness. Fourteen NW and 14 Ob individuals completed two trials in a randomized counterbalanced fashion, including a prior exercise trial (EX) and a no exercise trial (NoEX). During the EX trial, the participants performed 1 h of treadmill walking (55-60% peak O2 uptake) during the evening, 12 h before a 4-h standardized mixed meal test. Frequent blood samples were taken and analyzed for acylated ghrelin, and a visual analog scale was used to assess perceived hunger and fullness. In NW individuals, EX, compared with NoEX, reduced fasting acylated ghrelin concentrations by 18% (P = 0.03), and, in response to feeding, the change in acylated ghrelin (P = 0.02) was attenuated by 39%, but perceived hunger and fullness were unaltered. In Ob individuals, despite no changes in fasting or postprandial acylated ghrelin concentrations with EX, postprandial fullness was attenuated by 46% compared with NoEX (P = 0.05). In summary, exercise performed the night before a meal suppresses acylated ghrelin concentrations in NW individuals without altering perceived hunger or fullness. In Ob individuals, despite no changes in acylated ghrelin concentrations, EX reduced the fullness response to the test meal. Acylated ghrelin and perceived fullness responses are differently altered by acute aerobic exercise in NW and Ob individuals.


Managing to improve quality: the relationship between accreditation standards, safety practices, and patient outcomes.
Given the trend toward eliminating reimbursement for "never events," hospital administrators are challenged to implement practices designed to prevent their occurrence. Little evidence exists, however, that patient safety practices, as evaluated using accreditation criteria, are related to the achievement of patient safety outcomes. The aim of this study was to examine the relationship between patient safety practices, as measured by accreditation standards, and patient safety outcomes as measured by hospital rates of infections, decubitus ulcers, postoperative respiratory failure, and failure to rescue. Secondary data were used to examine relationships between patient-safety-related accreditation standards and patient outcomes in U.S. acute care hospitals. Accreditation performance areas were reduced into subscores to represent patient safety practices. Outcome rates were calculated using the Agency for Healthcare Research and Quality Patient Safety Indicator software. Multivariate regression was performed to determine the significance of the relationships. Three of four multivariate models significantly explained variance in hospital patient safety indicator rates. Accreditation standards reflecting patient safety practices were related to some outcomes but not others. Rates of infections and decubitus ulcers occurred more frequently in hospitals with poorer performance in utilizing patient safety practices, but no differences were noted in rates of postoperative respiratory failure or failure to rescue. Certain adverse events, such as infections and decubiti, may be reduced by preventive protocols that are reflected in accreditation standards, whereas other events, such as failure to rescue and postoperative respiratory failure, may require multifaceted strategies that are less easily translated into protocols. Our approach may have influenced the observed associations yet represents progress toward assessing whether safety practices, as measured by accreditation standards, are related to patient outcomes.


Pax 2 expression in mesodermal segmentation and its relationship with EphA4 and Lunatic-fringe during chicken somitogenesis.
In the Pax gene family, which encodes DNA-binding proteins, Pax 2 has been known to play important roles in the formation of the midbrain/hindbrain boundary, eye, inner ear and kidney in vertebrates (Bioessays 19 (1997) 755). In this article, we report a segmentally regulated pattern of Pax 2 expression during chicken somitogenesis. Pax 2 mRNA is localized in the rostral end of the unsegmented presomitic mesoderm (PSM), abutting anteriorly on a prospective segmentation border. This pattern repeats every segmentation cycle (90 min) observed in ovo and also in the half embryo culture assay in which one half of PSM along the midline is fixed immediately while the other half is cultured for a given period. We also determined the sequence of changes in Pax 2 expression during a segmentation cycle by comparing the pattern of Pax 2 with that of Lunatic-fringe (L-fringe), known to cycle periodically in posterior PSM. A systematic comparison of the expression patterns between Pax 2, L-fringe and EphA4 further highlighted a close relationship between EphA4 and Pax 2 during a segmentation cycle. Lastly, Pax 2 is not segmentally expressed in mouse PSM, suggestive of species (avian)-specific mechanisms underlying somitic segmentation.


[Prevalence of depressive disorders in children and adolescents attending primary care. A survey with the Aquitaine Sentinelle Network].
Since depressive disorders in children and adolescents have not been widely studied in the context of gene-ral medicine, we conducted an epidemiological survey among general practitioners (GP's) consulted by young subjects aged 7 to 17 years for various reasons. The aims were the following: to estimate the prevalence of depressive disorders in general practice, to detect the eventual existence of particular clinical forms, to assess the frequency of comorbid disorders and to determine to what degree these disorders were diagnosed by GP's. The study was conducted over 6 months in concert with 45 practitioners of the Aquitaine Sentinelle Network because of their strong experience in the field of epidemiological surveys, especially regarding psychiatric disorders. The population included all consecutive attenders aged 7 to 17 years. Consent to participate was obtained from children and adolescents and their parents. Finally 155 patients took part. A two-stage epidemiologic strategy was used, including screening tests in the first stage and semi structured interview by clinician in the second stage for diagnostic confirmation. During the first stage, information was obtained from children and adolescents and general practitioners using three questionnaires. The self-report questionnaire Center for Epidemiological Studies Depression (CES-D) was used for screening depression in 13 to 17 years old adolescents and the 20 items of the scale were modified to make it more comprehensible and relevant for children aged 7 to 12. The cut-off of 21 used in France appeared to be the more appropriate in both males and females and was taken to indicate high likelihood of depressive disorder. Therefore people with score 21 or more were approached for the second stage. The Child Behavior Checklist (CBCL), an instrument of well-established validity and reliability, provided information from parents about the child's behavior and competencies. Demographic and environmental data, as well as the reason for the visit and the presence of associated psychological factors were collected from a questionnaire devised for the study and completed by the practitioner. The 21 patients initially detected were invited to take part in the second stage. A total of 18 agreed to meet the psychiatrist. Sex-ratio female/male of this sample was 1,25 and mean age was 12,5 years. All of them underwent the Schedule for Affective Disorders and Schizophrenia for School Aged Children (Kiddie-SADS), a semi structured research interview of established validity. Diagnoses were made according to the DSM IV criteria (American Psychiatric Association). Results showed that more than one child out of 10 aged less than 13 years had a depressive disorder, and that the prevalence in the adolescent sub-group was 5%. Major depressive episode was present in 6% of the children sample, dysthymia in 4% and maladjustment disorder with depressive mood in about 1%. All depressive disorders were moderate. Atypical depression (in the Anglosaxon sense of the term) was present in half of the depressed adolescents. Other disorders included anxiety disorders with a rate of about 4% overanxious in the adolescent sample, obsessive compulsive disorder, panic disorder. Disruptive disorders were considerably less common. Psychiatric comorbidity, usually involving different types of emotional disorders, was present in about 50% of psychiatric cases, with a prevalence of anxiety disorders. The reasons why depressed subjects consulted were not specific. The most common reasons for visiting the GP were the somatic complaints with a rate of 50% in both populations, whatever the CES-D's score was. A few per cent of patients attending primary care presented with mental health complaints, and the rate was similar in the two populations. Frequency of consultation was not a discriminant factor of depression. Familial cohesion and school performance were not associated with the CES-D's score, nor familial psychiatric history. Personal psychiatric history was related to depression, whereas the occurrence of bereavement made the CES-D score positive but was not significantly associated with fully- blown depression. Finally, we estimated that 70% of diagnoses of depression were not made during the consultation with GP's. No particular characteristic of depressed children consulting GP's could be established. These findings underline the importance of training GP's in the screening of depressive disorders in children and adolescents. A better knowledge that young general practice attenders have high rates of depressive disorders may facilitate more rapid referral for psychiatric assessment and treatment.


Early changes in carcinogenesis of colorectal adenomas.
Adenocarcinoma of the colon and rectum is the third most common cause of cancer deaths and the sixth most common cancer in the world. Adenomas are benign neoplastic lesions which can be transformed into carcinomas, but this is usually not the case. There should be some risk factors which lead to the development of carcinomas into adenomas. The aim of this study is to find out the early changes and high risk factors related to carcinogenesis in colonic polyps. In this study, we reviewed nearly 1000 colonoscopic biopsies and chose 72 biopsies. We developed three groups (tubular adenomas group 1, villous adenomas group 2, normal mucosa group 3); each group had 24 different biopsies. P53, Ki-67, bcl-2, cyclin D1, E-cadherin, c-erb B2 immunohistochemistry and human papillomavirus (HPV) in-situ hybridization were used for analysis. Five of the seventy-two cases were positive in HPV in-situ analysis. Four of them were villous adenomas and one was a tubular adenoma. Ki-67 expression was limited only to crypts in group 3 but in groups 1 and 2, Ki-67 expression was seen both in crypt epithelium and surface epithelium. Cyclin D1, c-erb B2, bcl-2 expression was significantly increased in neoplastic polyps. Ki-67 expression, both in the crypt and surface epithelium, and cyclin D1, c-erb B2, bcl-2 over-expression may be a clue of dysplastic epithelium and if the role of HPV is elucidated and shown to be important in colonic carcinogenesis, then vaccination might prevent carcinogenesis caused by HPV.


Relationship between lysosomal membrane destabilization and chemical body burden in eastern oysters (Crassostrea virginica) from Galveston Bay, Texas, USA.
Lysosomal destabilization was measured by using hemocytes of eastern oysters (Crassostrea virginica) collected along a chemical concentration gradient in Galveston Bay, Texas, USA. Results of the lysosomal response were compared to concentrations of organic compounds and trace elements in oyster tissue. Concentrations (on a dry-wt basis) ranged from 288 to 2,390 ng/g for polycyclic aromatic hydrocarbons (PAHs), 38 to 877 ng Sn/g for tri-n-butyltin (TBT), 60 to 562 ng/g for polyclorinated biphenyls (PCBs), and 7 to 71 ng/g for total DDT. Trace element concentrations (on a dry-wt basis) ranged from 1.1 to 4.0 microg/g for Cd, 105 to 229 microg/g for Cu, 212 to 868 microg/g for Al, and 1,200 to 8,180 microg/g for Zn. The percentage of destabilized lysosomes ranged from 34 to 81%. A significant positive correlation (p < 0.05) was observed between lysosomal destabilization and body burden of organic compounds (PAHs, PCBs, TBT, and chlorinated pesticides). No significant correlation was found between metal concentrations and lysosomal destabilization. Based on lysosomal destabilization, the study sites in Galveston Bay can be placed in one of three groups: healthy (Hanna Reef and Confederate Bay), moderately damaged (Offats Bayou and Todd's Dump), and highly damaged (Yacht Club and Ship Channel). Lysosomal destabilization that is consistent with toxic chemical body burdens supports previous observations that lysosomal membranes are damaged by toxic chemicals and indicates that this method can serve as an early screening tool to assess overall ecosystem health by using oysters.


Closing the equity gap as we move to the elimination of seclusion: Early results from a national quality improvement project.
Use of seclusion within mental health inpatient facilities is harmful for consumers and staff, but it is still used in many Aotearoa New Zealand and Australian facilities, at higher, inequitable rates for the indigenous populations of both countries. We report early results from a national programme to eliminate seclusion in mental health services in Aotearoa New Zealand, using a bicultural approach to reduce inequity for Māori. The 'Zero Seclusion: Safety and dignity for all' programme, with programme teams nationwide, developed a co-designed bicultural change package combining Māori cultural and Western clinical interventions, incorporating quality improvement methodologies. Outcome measures included seclusion rates, duration, and average number of episodes per person admitted, by ethnicity, with a focus on equity. Nationally, rates of seclusion for Māori reduced from the 12-month (to August 2019) baseline mean monthly rate of 7.5% to 6.8%, sustained from late 2020 to September 2022. The duration of seclusion for Māori reduced by 33% (4.5 h at baseline to 3.0). A focus on inequity for Māori in use of seclusion, and a bicultural approach with cultural and clinical interventions, has been associated with a national reduction in rates and duration of seclusion for Māori.


Reduction of visual acuity decreases capacity to evaluate radiographic image quality.
To determine the impact of reduced visual acuity on the evaluation of a test object and appendicular radiographs. Visual acuity was reduced by two different magnitudes using simulation glasses and compared to normal vision (no glasses). During phase one phantom images were produced for the purpose of counting objects by 13 observers and on phase 2 image appraisal of anatomical structures was performed on anonymized radiographic images by 7 observers. The monitors were calibrated (SMPTE RP133 test pattern) and the room lighting was maintained at 7 ± 1 lux. Image display and data on grading were managed using ViewDEX (v.2.0) and the area under the visual grading characteristic (AUCVGC) was calculated using VGC Analyzer (v1.0.2). Inferential statistics were calculated using SPSS. For the evaluation of appendicular radiographs the total interpretation time was longer when visual acuity was reduced with 2 pairs of simulation glasses (15.4 versus 8.9 min). Visual grading analysis showed that observers can lose the ability to detect anatomical and contrast differences when they have a simulated visual acuity reduction, being more challenging to differentiate low contrast details. No simulation glasses, compared to 1 pair gives an AUCVGC of 0.302 (0.280, 0.333), that decreases to 0.197 (0.175, 0.223) when using 2 pairs of glasses. Reduced visual acuity has a significant negative impact on the evaluation of test objects and clinical images. Further work is required to test the impact of reduced visual acuity on visual search, technical evaluation of a wider range of images as well as pathology detection/characterization performance. It seems that visual performance needs to be considered to reduce the risks associated with incomplete or incorrect diagnosis. If employers or professional bodies were to introduce regular eye tests into health screening it may reduce the risk of misinterpretation as a result of poor vision.


A Comparative Analysis of the Risk Profiles of Australian Young Offenders From Rural and Urban Communities.
Australian young people from rural areas, particularly Aboriginal young people, are overrepresented in the juvenile justice system. Apart from broad evidence regarding the entrenched social disadvantages experienced by young people in rural communities, the literature is limited in describing why this might be case. Due to these social disadvantages, it is hypothesised that young offenders from rural communities will have higher levels of offending risk factors, as measured by the Youth Level of Service/Case Management Inventory-Australian Adaption (YLS/CMI-AA). A total of 6,750 archival records were analysed, showing that significantly more Aboriginal young offenders live in rural areas. Contrary to the hypothesis, urban young offenders had significantly higher risk scores than rural young offenders. These findings suggest that there may be particular ecological factors that are not assessed in the current risk assessment instrument or that rural young people have a range of protective factors that may insulate against the broader context of social disadvantage.


Use of Actuator-Driven Pulsed Water Jet in Brain and Spinal Cord Cavernous Malformations Resection.
A piezo actuator-driven pulsed water jet (ADPJ) system is a novel surgical instrument that enables dissection of tissue without thermal damage. It can potentially resect intra-axial lesions while preserving neurological function. To report our first experience of applying an ADPJ system to brain and spinal cord cavernous malformations. Four patients (2 women and 2 men, mean age 44.5 years) with brain (n = 3) and spinal cord (n = 1) cavernous malformations were enrolled in the study. All surgeries were performed with the aid of the ADPJ system. Postoperative neurological function and radiological findings were evaluated. The ADPJ system was useful in dissecting boundaries between the lesion and surrounding brain/spinal cord tissues. The pulsed water jet provided a clear surgical view and helped surgeons follow the margins. Water jet dissection peeled off the brain and spinal cord tissues from the lesion wall. Surrounding gliotic tissue was preserved. As a consequence, the cavernous malformations were successfully removed. Postoperative magnetic resonance imaging confirmed total removal of lesions in all cases. Preoperative neurological symptoms completely resolved in 2 patients. The others experienced partial recovery. No patients developed new postoperative neurological deficits; facial palsy temporarily worsened in 1 patient who underwent a suprafacial colliculus approach for the brainstem lesion. The ADPJ provided a clear surgical field and enabled surgeons to dissect boundaries between lesions and surrounding brain and spinal cord gliotic tissue. The ADPJ system is a feasible option for cavernous malformation surgery, enabling successful tumor removal and preservation of neurological function.


P53 mutation in acute T cell lymphoblastic leukemia is of somatic origin and is stable during establishment of T cell acute lymphoblastic leukemia cell lines.
Samples donated by patients with T cell acute lymphoblastic leukemia (T-ALL) were screened for mutations of the p53 tumor suppressor gene. Peripheral blood cells of T-ALL relapse patient H.A. were found to possess a heterozygous point mutation at codon 175 of the p53 gene. To determine whether this was an inherited mutation, a B cell line (HABL) was established. Leukemic T cell lines (HATL) were concurrently established by growing peripheral blood leukemic T cells at low oxygen tension in medium supplemented with IGF-I. Previously we had shown that > 60% of leukemic T cell lines possessed mutations in the p53 gene (Cheng, J., and M. Hass. 1990. Mol. Cell. Biol. 10:5502), mutations that might have originated with the donor's leukemic cells, or might have been induced during establishment of the cell lines. To answer whether establishment of the HATL lines was associated with the induction of p53 mutations, cDNAs of the HATL and HABL lines were sequenced. The HATL lines retained the same heterozygous p53 mutation that was present in the patient's leukemic cells. The HABL line lacked p53 mutations. Immunoprecipitation with specific anti-p53 antibodies showed that HATL cells produced p53 proteins of mutant and wild type immunophenotype, while the HABL line synthesized only wild-type p53 protein. The HATL cells had an abnormal karyotype, while the HABL cells possessed a normal diploid karyotype. These experiments suggest that (a) p53 mutation occurred in the leukemic cells of relapse T-ALL patient HA; (b) the mutation was of somatic rather than hereditary origin; (c) the mutation was leukemia associated; and (d) establishment of human leukemia cell lines needs not be associated with in vitro induction of p53 mutations. It may be significant that patient HA belonged to a category of relapse T-ALL patients in whom a second remission could not be induced.


Rapid 4 to 6 hour detection of extended-spectrum beta-lactamases in a routine laboratory.
With the growing frequency of extended-spectrum beta-lactamases (ESBL) among Enterobacteriaceae, treatment of Gram-negative nosocomial infections requires rapid and reliable detection of this enzyme. Quicolor agar (QC agar) (Salubris Inc., Massachusetts, USA) is a novel chromogenic agar medium changing colour within 4 to 6 h due to the metabolic activity of growing bacteria. This study investigated the use of QC agar compared to Mueller Hinton agar (MH) for the detection of ESBL using disk diffusion and E-test. 100 Enterobacteriaceae isolated at Hacettepe University Hospital, of which 50 were predetermined to be ESBL positive and 50 as negative using the CLSI disk diffusion ESBL (phenotypic confirmatory test) criteria. For disk diffusion and E-test, cefotaxime+/-clavulanate (CT/CTL) and ceftazidime+/-clavulanate (TZ/TZL) were used, and for E-test, cefepime+/-clavulanate (PM/PML) was also used. QC agar rapid ESBL results for all strains were in agreement with the standard overnight procedure. All 50 ESBL positives were detected by both methods. For the 50 ESBL negatives, QC agar rapid results from E-test and disk diffusion were in complete accordance with the overnight MH results. Moreover, E-test detected 8 additional ESBL positive strains that disk diffusion missed. For disk diffusion, CT/CTL alone detected all 50 ESBL positives while TZ/TZL alone missed 5 ESBL positives. E-test CT/CTL alone confirmed all 50 ESBL positives and identified 4 additional ESBL-positive strains. When used together, E-test CT/CTL, TZ/TZL and PM/PML identified a total of 58 ESBL positives among the 100 strains tested. QC agar can be used for rapid and reliable ESBL detection within 4 to 6 h, using disk diffusion and E-test ESBL reagents. This rapid method should be further validated using genotype characterized ESBL and other beta-lactamase positive strains.


A C-terminal domain directs Kv3.3 channels to dendrites.
Pyramidal neurons of the electrosensory lateral line lobe (ELL) of Apteronotus leptorhynchus express Kv3-type voltage-gated potassium channels that give rise to high-threshold currents at the somatic and dendritic levels. Two members of the Kv3 channel family, AptKv3.1 and AptKv3.3, are coexpressed in these neurons. AptKv3.3 channels are expressed at uniformly high levels in each of four ELL segments, whereas AptKv3.1 channels appear to be expressed in a graded manner with higher levels of expression in segments that process high-frequency electrosensory signals. Immunohistochemical and recombinant channel expression studies show a differential distribution of these two channels in the dendrites of ELL pyramidal neurons. AptKv3.1 is concentrated in somas and proximal dendrites, whereas AptKv3.3 is distributed throughout the full extent of the large dendritic tree. Recombinant channel expression of AptKv3 channels through in vivo viral injections allowed directed retargeting of AptKv3 subtypes over the somadendritic axis, revealing that the sequence responsible for targeting channels to distal dendrites lies within the C-terminal domain of the AptKv3.3 protein. The targeting domain includes a consensus sequence predicted to bind to a PDZ (postsynaptic density-95/Discs large/zona occludens-1)-type protein-protein interaction motif. These findings reveal that different functional roles for Kv3 potassium channels at the somatic and dendritic level of a sensory neuron are attained through specific targeting that selectively distributes Kv3.3 channels to the dendritic compartment.


Characterization of antiviral activity of entecavir in transgenic mice expressing hepatitis B virus.
Entecavir (ETV), a cyclopentyl guanosine nucleoside analog, was evaluated in transgenic mice expressing hepatitis B virus (HBV). ETV administered orally once daily for 10 days at a dosage of 3.2mg/kg significantly (P<or=0.001) reduced liver HBV DNA in female mice from 5.9 to <0.82 pg of HBV DNA per microg of cellular DNA, and from 8.3 to <1.1 pg/microg in male mice. To compare the efficacy of ETV with other compounds previously evaluated in this model and with ETV activities in other animal models, the efficacy of serial one-half log dilutions of ETV were evaluated in both male and female mice to determine the minimal effective dose. End-point titration experiments resulted in a statistically significant HBV DNA reduction in the liver at concentrations of 0.032 and 0.1mg/kg per day in female and male mice, respectively. Viral liver RNA, and serum e (HBeAg), serum surface (HBsAg), and liver core antigens (HBcAg) were not affected by ETV treatment presumably because the antiviral target was viral polymerase activity and the HBV produced from the transgene was not capable of secondary rounds of infection in the mouse. ETV was well tolerated and no morbidity or mortality was observed during the 10-day study. Similar to other animal models, ETV displayed potent anti-HBV activity in this transgenic mouse model.


Optimal design of multiple-channel RF pulses under strict power and SAR constraints.
Parallel radio frequency transmission has recently been explored as a means of tailoring the spatial response of MR excitation. In particular, parallel transmission is increasingly used to accelerate radio frequency pulses that rely on time-varying gradient fields to achieve selectivity in multiple dimensions. The design of the underlying multiple-channel radio frequency waveforms is mostly based on regularized least-squares optimization in close analogy with image reconstruction in parallel imaging. However, this analogy has important limitations. Unlike image reconstruction, the design of radio frequency waveforms is subject to multiple strict constraints, which arise from technical power limits, as well as safety limits on local and global energy deposition in vivo. To optimize excitation profiles under such strict constraints, it is proposed to depart from the regularization strategy and rely on semidefinite programming instead. To render this approach fast, it is performed in a reduced search space, which is obtained by initial Lanczos iteration. The proposed algorithm is demonstrated to enable efficient pulse optimization within exactly the given constraints, including local specific absorption rate limits for multiple compartments. It is also shown that the proposed approach readily accommodates advanced forward models of the excitation process, including the effects of local off-resonance and transverse relaxation.


Structural analysis of Sindbis virus capsid mutants involving assembly and catalysis.
Sindbis virus core protein (SCP) has been isolated from virus and crystallized. The X-ray crystallographic structure showed that the amino-terminal 113 residues appeared to be either disordered or truncated during crystallization and that the carboxy-terminal residues 114 to 264 had a chymotrypsin-like structure. The carboxy-terminal residues 106 to 264 and 106 to 266 of SCP have now been expressed in Escherichia coli. Most crystal forms of the truncated proteins were isomorphous with those of the virally extracted protein. There are only small structural differences between the truncated recombinant protein and the ordered part of the wild-type virus-extracted protein. Hence, E. coli-expressed SCP can be used to study proteolytic properties and the contribution of SCP to nucleocapsid assembly, interaction with the E2 glycoprotein and interaction with RNA. The same dimer that was found in two different crystal forms of the virus-extracted SCP was present also in some of the crystals of the truncated recombinant protein. The monomer-monomer interface is maintained by two pairs of hydrogen bonds and by hydrophobic interactions. Removal of the hydrogen bonds by single substitutions did not prevent dimer formation. However, a mutation that reduced the hydrophobic contacts did inhibit dimer formation. The wild-type truncated SCP is active in E. coli, as evidenced by proteolytic processing of a series of progressively longer precursors that extend beyond residue 264. Unlike the virus-extracted capsid protein, the E. coli-expressed SCP described here is terminated following the carboxy-terminal residue and, therefore, does not require autocatalysis. Nevertheless, the E. coli-expressed protein folds with the carboxy-terminal tryptophan residue in the specificity pocket. Two crystallographically independent molecules of SCP(106 to 266), which had two additional downstream residues and had the essential S215 mutated to alanine, showed two distinct modes of binding the uncleaved carboxy-terminal residues. These may represent successive steps of binding substrate prior to catalytic cleavage. Refinement of the various crystal structures of SCP showed that the amino-terminal arm from residues 107 to 113 was not disordered, but is associated with neighboring molecules. Residues 108 to 111 bind into a hydrophobic pocket composed primarily of Y180, W247 and F166. It had been shown that the double mutant (Y180S; E183G), with the Y180S substitution in this pocket, produced a large number of non-infectious virions, possibly because of modification in the interaction of the glycoprotein spikes with core proteins. The crystal structure of this double mutant showed that there was a large positional change in the side-chain of W247, which moved into the space created by the replacement of Y180 with serine. These conformational changes may alter the stability of the virion and, thus, regulate its functional requirements during cell entry.


End of an era of administering erythropoiesis stimulating agents among Veterans Administration cancer patients with chemotherapy-induced anemia.
Erythropoisis stimulating agent (ESA) use was addressed in Food and Drug Administration (FDA) Oncology Drug Advisory Committee (ODAC) meetings between 2004 and 2008. FDA safety-focused regulatory actions occurred in 2007 and 2008. In 2007, black box warnings advised of early death and venous thromboembolism (VTE) risks with ESAs in oncology. In 2010, a Risk Evaluation Strategies (REMS) was initiated, with cancer patient consent that mortality and VTE risks were noted with ESAs. We report warnings and REMS impacts on ESA utilization among Veterans Administration (VA) cancer patients with chemotherapy-induced anemia (CIA). Data were from Veterans Affairs database (2003-2012). Epoetin and darbepoetin use were primary outcomes. Segmented linear regression was used to estimate changes in ESA use levels and trends, clinical appropriateness, and adverse events (VTEs) among chemotherapy-treated cancer patients. To estimate changes in level of drug prescription rate after policy actions, model-specific indicator variables as covariates based on specific actions were included. ESA use fell by 95% and 90% from 2005, for epoetin and darbepoetin, from 22% and 11%, respectively, to 1% and 1%, respectively, among cancer patients with CIA, respectively (p<0.01). Following REMS in 2010, mean hematocrit levels at ESA initiation decreased from 30% to 21% (p<0.01). Black box warnings preceded decreased ESA use among VA cancer patients with CIA. REMS was followed by reduced hematocrit levels at ESA initiation. Our findings contrast with privately- insured and Medicaid insured cancer patient data on chemotherapy-induced anemia where ESA use decreased to 3% to 7% by 2010-2012. By 2012, the era of ESA administration to VA to cancer patients had ended but the warnings remain relevant and significant. In 2019, oncology/hematology national guidelines (ASCO/ASH) recommend that cancer patients with chemotherapy-induced anemia should receive ESAs or red blood cell transfusions after risk-benefit evaluation.


A Human Endogenous Bornavirus-Like Nucleoprotein Encodes a Mitochondrial Protein Associated with Cell Viability.
Endogenous retroviruses (ERVs) are sequences in animal genomes that originated from ancient retrovirus infections; they provide genetic novelty in hosts by being coopted as functional genes or elements during evolution. Recently, we demonstrated that endogenous elements from not only from retroviruses but also nonretroviral RNA viruses are a possible source of functional genes in host animals. The remnants of ancient bornavirus infections, called endogenous bornavirus-like elements (EBLs), are present in the genomes of a wide variety of vertebrate species, and some express functional products in host cells. Previous studies have predicted that the human EBL locus derived from bornavirus nucleoprotein, termed hsEBLN-2, expresses mRNA encoding a protein, suggesting that hsEBLN-2 has acquired a cellular function during evolution. However, the detailed function of the hsEBLN-2-derived product remains to be elucidated. In this study, we show that the hsEBLN-2-derived protein E2 acts as a mitochondrial protein that interacts with mitochondrial host factors associated with apoptosis, such as HAX-1. We also demonstrate that knockdown of hsEBLN-2-derived RNA increased the levels of PARP and caspase-3 cleavage and markedly decreased cell viability. In contrast, overexpression of E2 enhanced cell viability, as well as the intracellular stability of HAX-1, under stress conditions. Our results suggest that hsEBLN-2 has been coopted as a host gene, the product of which is involved in cell viability by interacting with mitochondrial proteins. IMPORTANCE Our genomes contain molecular fossils of ancient viruses, called endogenous virus elements (EVEs). Mounting evidence suggests that EVEs derived from nonretroviral RNA viruses have acquired functions in host cells during evolution. Previous studies have revealed that a locus encoding a bornavirus-derived EVE, hsEBLN-2, which was generated approximately 43 million years ago in a human ancestor, may be linked to the development of some tumors. However, the function of hsEBLN-2 has not been determined. In this study, we found that the E2 protein, an expression product of hsEBLN-2, interacts with apoptosis-related host proteins as a mitochondrial protein and affects cell viability. This study suggests that nonretroviral RNA viral EVEs have been coopted by hosts with more diverse functions than previously thought, showing a pivotal role for RNA virus infection in evolution.


CD226 (DNAM-1) is associated with susceptibility to juvenile idiopathic arthritis.
Juvenile idiopathic arthritis (JIA) is considered a complex genetic autoimmune disease. We investigated the association of genetic variants previously implicated in JIA, autoimmunity and/or immunoregulation, with susceptibility to JIA. A genetic association study was performed in 639 JIA patients and 1613 healthy controls of northwest European descent. Ninety-three single nucleotide polymorphisms (SNP) were genotyped in a candidate gene approach. Results of the entire JIA patient group (all subtypes) were compared with results obtained, alternatively, with a clinically homogeneous patient group including only oligoarticular and rheumatoid factor (RF) negative polyarticular JIA patients (n=493). Meta-analyses were performed for all SNPs that have been typed in other Caucasian JIA cohorts before. SNPs in or near PTPN22, VTCN1, the IL2-IL21 region, ANKRD55 and TNFA were confirmed to be associated with JIA (p<0.05), strengthening the evidence for involvement of these genes in JIA. In the majority of these replicated SNPs, effect sizes were larger when analysing a homogeneous patient cohort than when analysing all subtypes. We identified two novel associations with oligoarticular and RF-negative polyarticular JIA: CD226 rs763361 (OR 1.30, 95% CI 1.12 to 1.51, p=0.0006) and CD28 rs1980422 (OR 1.29, 95% CI 1.07 to 1.55, p=0.008). Meta-analyses including reported studies confirmed the association of both SNPs with susceptibility to JIA (OR 1.16, p=0.001 and OR 1.18, p=0.001, for rs763361 and rs1980422, respectively). The CD226 gene has been identified as novel association with JIA, and a SNP near CD28 as a suggestive association. Both genes are probable candidate risk factors, since they are involved in costimulation of T cells.


How large are the nonspecific effects of acupuncture? A meta-analysis of randomized controlled trials.
While several recent large randomized trials found clinically relevant effects of acupuncture over no treatment or routine care, blinded trials comparing acupuncture to sham interventions often reported only minor or no differences. This raises the question whether (sham) acupuncture is associated with particularly potent nonspecific effects. We aimed to investigate the size of nonspecific effects associated with acupuncture interventions. MEDLINE, Embase, Cochrane Central Register of Controlled Clinical Trials and reference lists were searched up to April 2010 to identify randomized trials of acupuncture for any condition, including both sham and no acupuncture control groups. Data were extracted by one reviewer and verified by a second. Pooled standardized mean differences were calculated using a random effects model with the inverse variance method. Thirty-seven trials with a total of 5754 patients met the inclusion criteria. The included studies varied strongly regarding patients, interventions, outcome measures, methodological quality and effect sizes reported. Among the 32 trials reporting a continuous outcome measure, the random effects standardized mean difference between sham acupuncture and no acupuncture groups was -0.45 (95% confidence interval, -0.57, -0.34; I2 = 54%; Egger's test for funnel plot asymmetry, P = 0.25). Trials with larger effects of sham over no acupuncture reported smaller effects of acupuncture over sham intervention than trials with smaller nonspecific effects (β = -0.39, P = 0.029). Sham acupuncture interventions are often associated with moderately large nonspecific effects which could make it difficult to detect small additional specific effects. Compared to inert placebo interventions, effects associated with sham acupuncture might be larger, which would have considerable implications for the design and interpretation of clinical trials.


Rebound Effects Caused by Withdrawal of MET Kinase Inhibitor Are Quenched by a MET Therapeutic Antibody.
MET oncogene amplification is emerging as a major mechanism of acquired resistance to EGFR-directed therapy in lung and colorectal cancers. Furthermore, MET amplification predicts responsiveness to MET inhibitors currently in clinical trials. Among the anti-MET drugs available, ATP-competitive small-molecule kinase inhibitors abrogate receptor autophosphorylation and downstream activation of ERK1/2 and AKT, resulting in cell-cycle arrest. However, this antiproliferative effect allows persistence of a pool of cancer cells that are quiescent but alive. Once the inhibition is removed, rebound activation of MET-driven cell proliferative pathways and tumor growth may occur, an adverse event observed frequently in clinical settings after drug discontinuation. Here we show that inhibitor withdrawal prompts receptor phosphorylation to levels higher than those displayed at steady-state and generates a rebound effect pushing quiescent cancer cells back into the cell cycle, both in vitro and in experimental tumor models in vivo Mechanistically, we found that inhibitor treatment blocks MET endocytosis, causing a local increase in the number of receptors at the plasma membrane. Upon inhibitor washout, the receptor is readily rephosphorylated. The initial phosphorylation is not only increased but also prolonged in duration due to downmodulation of a phosphatase-mediated MET-negative feedback loop, which accompanies receptor internalization. Notably, treatment with a MET therapeutic antibody that induces proteolytic cleavage of the receptor at the cell surface substantially prevents this rebound effect, providing a rationale to combine or alternate these mechanistically different types of MET-targeted therapy. Cancer Res; 76(17); 5019-29. ©2016 AACR.


Contemporary cryptic sexuality in Trypanosoma cruzi.
Clonal propagation is considered to be the predominant mode of reproduction among many parasitic protozoa. However, this assumption may overlook unorthodox, infrequent or cryptic sexuality. Trypanosoma cruzi, which causes Chagas disease, is known to undergo non-Mendelian genetic exchange in the laboratory. In the field, evidence of extant genetic exchange is limited. In this study, we undertook intensive sampling of T. cruzi Discrete Typing Unit I in endemic eastern Colombia. Using Fluorescence-activated cell sorting, we generated 269 biological clones from 67 strains. Each clone was genotyped across 24 microsatellite loci. Subsequently, 100 representative clones were typed using 10 mitochondrial sequence targets (3.76 Kbp total). Clonal diversity among humans, reservoir hosts and vectors suggested complex patterns of superinfection and/or coinfection in oral and vector-borne Chagas disease cases. Clonal diversity between mother and foetus in a congenital case demonstrates that domestic TcI genotypes are infective in utero. Importantly, gross incongruence between nuclear and mitochondrial markers is strong evidence for widespread genetic exchange throughout the data set. Furthermore, a confirmed mosaic maxicircle sequence suggests intermolecular recombination between individuals as a further mechanism of genetic reassortment. Finally, robust dating based on mitochondrial DNA indicates that the emergence of a widespread domestic TcI clade that we now name TcI(DOM) (formerly TcIa/VEN(Dom)) occurred 23 000 ± 12 000 years ago and was followed by population expansion, broadly corresponding with the earliest human migration into the Americas.


Neuron-specific enolase concentrations in blood as a prognostic parameter in cerebrovascular diseases.
To investigate the clinical relevance of plasma concentrations of neuron-specific enolase (NSE) in patients with severe cerebrovascular diseases, serial analyses were performed during the first 10 days after the acute event. Plasma samples taken from 61 patients (30 with brain infarction, 13 with intracerebral hemorrhage, 11 with cardiogenic hypoxia-ischemia, and 7 with myocardial infarction [as control group]) were analyzed for NSE concentration using an enzyme immunoassay. The time course of plasma NSE was correlated with clinical findings, clinical outcome, cranial computed tomography, intracranial pressure, and other laboratory data. In cases of hypoxia-ischemia there was close correlation between plasma NSE values during the first 72 hours and the clinical outcome. In brain infarction and intracerebral hemorrhage, high plasma NSE mostly indicated an unfavorable outcome, but low values did not permit a reliable prognostic estimation. In cases of cerebral infarction and intracerebral hemorrhage with secondary neuronal destruction (for example, due to malignant edema), increasing NSE concentrations in plasma preceded the change of clinical or other diagnostic parameters. The course of plasma NSE levels is seen as a relevant parameter for assessing the prognosis of cerebral hypoxia-ischemia. Additionally, it may prove to be a useful tool for monitoring space-occupying brain infarctions and intracerebral hemorrhages and therefore may contribute to improved therapeutic management of severe cerebrovascular diseases.


Short- and long-term effects of conventional and artificial rearing strategies on the health and performance of growing lambs.
Artificial rearing of young animals represents a challenge in modern ruminant production systems. This work aims to evaluate the short- and long-term effects of the type of rearing on the animal's health, growth, feed utilization and carcass performance. A total of 24 pregnant ewes carrying triplets were used. Within each triplet set, lambs were randomly allocated to one experimental treatment: natural rearing on the ewe (NN); ewe colostrum for 24 h followed by artificial rearing with milk replacer (NA) and 50 g of colostrum alternative supplementation followed by artificial rearing (AA). Milk replacer, ryegrass hay and creep feed were offered ad libitum, and each experimental group was kept in independent pens until weaning at 45 days of age. After weaning all lambs were placed together on the same pasture for fattening for 4 months. Blood samples were taken at 24 h after birth, at weaning and at the end of the fattening period (23 weeks). Results showed that no failure in the passive immune transfer was detected across treatments. Although artificially reared lambs at weaning had lower plasma levels of β-hydroxy-butyrate (-62%), high-density lipoproteins (-13%) and amylase (-25%), and higher levels of low-density lipoproteins (+38%) and alkaline phosphatase (+30%), these differences disappeared during the fattening period. Only the greater levels of calcium and the lower levels of haemoglobin and white blood cells detected at weaning in artificially reared lambs (+7.2%, -2.8% and -17.8%) persisted by the end of the fattening period (+4.3%, -3.3% and -9.5%, respectively). Minor diarrheal events from weeks 2 to 5 were recorded with artificial rearing, leading to lower growth rates during the 1st month. However, these artificially reared lambs caught up towards the end of the milk feeding period and reached similar weaning weights to NN lambs. During the fattening period NN lambs had a greater growth rate (+16%) possibly as a result of their greater early rumen development, which allowed a higher feed digestibility during the fattening period in comparison to NA lambs (+5.9%). As a result, NN lambs had heavier final BWs (+7.0%), but tended to have lower dressing percentage (-5.7%) than artificially reared lambs, thus no differences were noted in either carcass weight or in carcass conformation across treatments. In conclusion, the use of a colostrum alternative and milk replacer facilitated the successful rearing of lambs, reaching similar productive parameters; however, special care must be taken to maximize the rumen development before weaning.


Bio-Inspired Balance Control Assistance Can Reduce Metabolic Energy Consumption in Human Walking.
The amount of research on developing exoskeletons for human gait assistance has been growing in the recent years. However, the control design of exoskeletons for assisting human walking remains unclear. This paper presents a novel bio-inspired reflex-based control for assisting human walking. In this approach, the leg force is used as a feedback signal to adjust hip compliance. The effects of modulating hip compliance on walking gait is investigated through joint kinematics, leg muscle activations and overall metabolic costs for eight healthy young subjects. Reduction in the average metabolic cost and muscle activation are achieved with fixed hip compliance. Compared to the fixed hip compliance, improved assistance as reflected in more consistent reduction in muscle activities and more natural kinematic behaviour are obtained using the leg force feedback. Furthermore, smoother motor torques and less peak power are two additional advantages obtained by compliance modulation. The results show that the proposed control method which is inspired by human posture control can not only facilitate the human gait, but also reduce the exoskeleton power consumption. This demonstrates that the proposed bio-inspired controller allows a synergistic interaction between human and robot.


Is preschool executive function causally related to academic achievement?
The primary objective of this study was to reevaluate the well-established result that preschoolers' performance on executive function tasks are positively associated with their performance on academic achievement tests. The current study replicated the previously established concurrent associations between children's performance on EF tasks and academic achievement tests. Specifically, children's performance on measures of inhibitory and motor control were positively associated with their performance on tests of reading, writing, and mathematics achievement (rs = .2-.5); moreover, although diminished in magnitude, most of these associations held up even after including an earlier measure of academic achievement as a covariate (rs = .1-.3). However, the application of an alternative analytic method, fixed effects analysis, a method that capitalizes on repeated measures data to control for all time stable measured and unmeasured covariates, rendered the apparent positive associations between executive function and academic achievement nonsignificant (rs = .0-.1). Taken together, these results suggest that the well-replicated association between executive function abilities and academic achievement may be spurious. Results are discussed with respect to the importance of utilizing analytic methods and research designs that facilitate strong causal inferences between executive function and academic achievement in early childhood, as well as the limitations of making curriculum development recommendations and/or public policy decisions based on studies that have failed to do so.


DNA and RNA-controlled switching of protein kinase activity.
Protein switches use the binding energy gained upon recognition of ligands to modulate the conformation and binding properties of protein segments. We explored whether the programmable nucleic acid mediated recognition might be used to design or mimic constraints that limit the conformational freedom of peptide segments. The aim was to design nucleic acid-peptide conjugates in which the peptide portion of the conjugate would change the affinity for a protein target upon hybridization. This approach was used to control the affinity of a PNA-phosphopeptide conjugate for the signal transduction protein Src kinase, which binds the cognate phosphopeptides in a linear conformation. Peptide-nucleic acid arms were attached to known peptide binders. The chimeric molecules were studied in three modes: 1) as single strands, 2) constrained by intermolecular hybridization (duplex formation) and 3) constrained by intramolecular hybridization (hairpin formation). Of note, duplexes that were designed to accommodate bulged peptide structures (for example, in hairpins or bulges) had lower binding affinities than duplexes in which the peptide was allowed to adopt a more relaxed conformation. Greater than 90-fold differences in binding affinities were observed. It was, thus, feasible to make use of DNA hybridization to reversibly switch from no to almost complete inhibition of Src-SH2-peptide binding, and vice versa. A series of DNA and PNA-based hybridization experiments revealed the importance of charges and conformational effects. Nucleic acid mediated switching was extended to the use of RNA; this enabled a regulation of the enzymatic activity of the Src kinase. The proof-of-principle results demonstrate for the first time that PNA-peptide chimeras can transduce changes of the concentration of a given RNA molecule to changes of the activity of a signal transduction enzyme.


The impact of common chronic conditions on health-related quality of life: a general population survey in Iran using EQ-5D-5L.
Diseases have undeniable effects on Health-Related Quality of Life (HRQoL). Chronic diseases, in particular, limit the productive potentials and HRQoL of individuals. EQ-5D is a very popular generic instrument, which can be used to estimate HRQoL scores in any diseases. The current study investigates mean HRQoL scores in certain chronic diseases and examines the relationship between utility scores and chronic diseases in Iran. This cross-sectional study was carried out among the general adult population of Tehran. 3060 individuals were chosen by a stratified probability sampling method. The EQ-5D-5L questionnaire was applied. The utility scores were estimated using the Iranian crosswalk-based value set. The effect of chronic diseases on the HRQoL scores was derived by the Ordinary Least Squares (OLS) method. Data was analyzed using Stata version 13 software. The mean ± standard deviation utility and EQ-VAS scores were 0.85 ± 0.14 and 76.73 ± 16.55 in the participants without any chronic conditions. The scores were 0.69 ± 0.17 and 61.14 ± 20.61 in the participants with chronic conditions. The highest and lowest mean utility scores were related to thyroid disease (0.70) and Stroke (0.54), respectively. Common chronic conditions had significant negative effects on the HRQoL scores. Stroke (0.204) and cancer (0.177) caused the most reduction in the EQ-5D-5L utility scores. Lumbar disc hernia, digestive diseases, osteoarthritis, breathing problems, and anxiety/nerves cause 0.133, 0.109, 0.108, 0.087, and 0.078 reductions, respectively, in the EQ-5D-5L utility scores. This study provides insight into some common chronic conditions and their effects on the HRQoL. Policymakers and planners should pay attention to the effects of chronic conditions especially high prevalence one. They should adopt effective interventions to control this issue and increase health. The results of this study can also be beneficial in economic evaluation studies.


Comparison of Cockcroft-Gault and modification of diet in renal disease formulas as predictors of cardiovascular outcomes in patients with myocardial infarction treated with primary percutaneous coronary intervention.
We prospectively assessed the value of estimated glomerular filtration rate (eGFR) by the Modification of Diet in Renal Disease (MDRD) and Cockcroft-Gault (C-G) equations in predicting inhospital adverse outcomes after primary coronary intervention for acute ST-segment elevation myocardial infarction. We classified 647 patients into 3 categories according to eGFR, <60, 60 to 90, and >90 mL/min/1.73 m(2). The eGFRC-G classified 17 patients in the >90 mL/min/1.73 m(2) subgroup and 6 and 11 patients in the 60 to 90 and <60 mL/min/1.73 m(2) subgroups, respectively. In multivariate analysis, patients with eGFRC-G < 60 mL/min/1.73 m(2) had 19.5-fold (95% confidence interval [CI] 1.55-178) higher mortality risk and 5.48-fold (95% CI 1.75-24.21) higher major adverse cardiac events risk compared to patients with eGFRC-G >90 mL/min/1.73 m(2) (P = .01 and P = .01, respectively); the eGFRMDRD was not predictive. Although the MDRD equation more accurately estimates GFR in certain populations, the CG formula may be a better predictor of adverse events.


Herpesvirus deconjugases inhibit the IFN response by promoting TRIM25 autoubiquitination and functional inactivation of the RIG-I signalosome.
The N-terminal domains of the herpesvirus large tegument proteins encode a conserved cysteine protease with ubiquitin- and NEDD8-specific deconjugase activity. The proteins are expressed during the productive virus cycle and are incorporated into infectious virus particles, being delivered to the target cells upon primary infection. Members of this viral enzyme family were shown to regulate different aspects of the virus life cycle and the innate anti-viral response. However, only few substrates have been identified and the mechanisms of these effects remain largely unknown. In order to gain insights on the substrates and signaling pathways targeted by the viral enzymes, we have used co-immunoprecipitation and mass spectrometry to identify cellular proteins that interact with the Epstein-Barr virus encoded homologue BPLF1. Several members of the 14-3-3-family of scaffold proteins were found amongst the top hits of the BPLF1 interactome, suggesting that, through this interaction, BPLF1 may regulate a variety of cellular signaling pathways. Analysis of the shared protein-interaction network revealed that BPLF1 promotes the assembly of a tri-molecular complex including, in addition to 14-3-3, the ubiquitin ligase TRIM25 that participates in the innate immune response via ubiquitination of cytosolic pattern recognition receptor, RIG-I. The involvement of BPLF1 in the regulation of this signaling pathway was confirmed by inhibition of the type-I IFN responses in cells transfected with a catalytically active BPLF1 N-terminal domain or expressing the endogenous protein upon reactivation of the productive virus cycle. We found that the active viral enzyme promotes the dimerization and autoubiquitination of TRIM25. Upon triggering of the IFN response, RIG-I is recruited to the complex but ubiquitination is severely impaired, which functionally inactivates the RIG-I signalosome. The capacity to bind to and functionally inactivate the RIG-I signalosome is shared by the homologues encoded by other human herpesviruses.


The CHANGE program: Exercise intervention in primary care.
Primary care settings require a feasible program for integrating lifestyle interventions, which can reverse metabolic abnormalities, for patients in practice. To integrate a lifestyle intervention program into existing primary care clinics with an interprofessional approach that includes dietitians and kinesiologists. Canadian Health Advanced by Nutrition and Graded Exercise (CHANGE) provides a personalized approach to nutrition and exercise modification focusing on patients with metabolic syndrome. With CHANGE, exercise intervention is individualized (ie, tailored to individual preferences) and graded (ie, intensity is built up slowly over time); supervision and implementation of the program is conducted in a collaborative fashion between the family physician and the kinesiologist. Patients undergo an initial fitness assessment that determines their baseline aerobic, strength, and flexibility scores, and the same assessment is performed at 3 months and at 12 months. The CHANGE program demonstrates how interprofessional primary care teams can support patients with metabolic syndrome in achieving their health goals. By including dietitians and kinesiologists in primary care settings to work alongside family doctors, many barriers to lifestyle interventions can be overcome. The team's collaborative understanding of the patient combined with the patient's own sense of urgency for change creates the opportunity for the formation of new healthy lifestyle habits. Although results are preliminary, CHANGE appears to be a feasible, implementable, and effective program.


General practice trainees face practice ownership with fear.
Objectives The aim of the present secondary analysis of data collected in a grounded theory study was to explore the perceptions of Registrars and new Fellows to practice ownership and management. Methods Focus groups and interviews with Registrars and recent Fellows were undertaken to explore the desire to become an owner, facilitators and barriers to practice ownership and delivery models for practice ownership education. A secondary thematic analysis was conducted to understand emerging concepts related to perceptions of general practice ownership. Results A surprisingly strong theme of fear was evident across focus group and interview participants. Expressed fear was specifically related to financial concerns, lack of relevant knowledge and skills and concern over balancing different roles. Moderating factors included previous life and educational experiences, as well as role modelling. Conclusions Graduation of a cohort of new general practitioners (GPs) who express fear towards practice ownership is concerning. Creating more positive learning environments and opportunity for open discussion regarding practice management and ownership is an important step in providing adequate support for new GPs to give serious consideration to career options. What is known about the topic? The traditional model of general practice ownership has been for a doctor to own and/or manage the practice. Fewer new GPs are taking on the role of owning a general practice, and disinterest has been presumed to play a significant role in this trend. It has been reported that current curricula provide insufficient focus on providing learning opportunities for general practice trainees on ownership and management models; however, recent research has shown that general practice trainees have a strong interest in receiving this knowledge during their training. What does this paper add? The present qualitative research evaluated the emotional response that general practice trainees (Registrars) expressed when they considered practice ownership. GP Registrars expressed fear when asked about the prospect of owning their own practice. This may be influenced by the role modelling they receive during their training. What are the implications for practitioners? Supervisors involved in general practice training should ensure their trainees have the opportunity to learn about practice ownership during their training. Supervisors should also be aware that their own behaviour and attitudes towards practice ownership is observed by their trainees and it can colour the emotional response the trainee has when considering owning a practice.


Copper removal from acid mine drainage-polluted water using glutaraldehyde-polyethyleneimine modified diatomaceous earth particles.
Mine waters and tailings generated from mining and mineral processing activities often have detrimental impact on the local environment. One example is acid mine drainage, in which sulphides in the mining waste react with water and oxygen to produce an acidic environment that subsequently dissolves host rock minerals from the waste containing toxic metals and trace elements. Copper is one such metal of significance, as it is mined at large volumes in sulphide containing ores. It has strong biocidal activity that greatly affects ecosystems. We have previously reported that glutaraldehyde (GA)-crosslinked polyethyleneimine (PEI) has strong affinity and selectivity for copper and that diatomaceous earth (DE) particles can be modified with the material to form a copper-extraction resin. In this study, the copper uptake of GA-PEI-DE particles was investigated from synthetic and real acid mine drainage samples under different pHs and their copper removal performance was compared with that of selected commercial resins. The results revealed that copper could effectively and preferentially bind to the material at pH 4, and that the copper could be completely eluted by lowering of the pH. In addition, effective copper uptake and elution was demonstrated using real legacy acid mine drainage water from Mount Lyell in Tasmania.


Financial incentives linked to self-assessment of prescribing patterns: a new approach for quality improvement of drug prescribing in primary care.
Financial incentives have been suggested to be effective in increasing the quality and efficiency of drug prescribing. Concern has been raised in relation to potential negative consequences on the quality of care. To describe and analyse the impact of an incentives model linking payment with adherence to drug and therapeutics committee (DTC) guidelines and self-reflection of prescribing pattern in a 'prescribing quality report'. The study was performed in the county of Stockholm, Sweden, with 139 (out of 154) primary healthcare centres (PHCs) participating in the project and 15 PHCs not participating. The study consisted of two parts: a quantitative observational study of prescribing patterns and a qualitative analysis of the submitted prescribing quality reports. All prescriptions issued from PHCs and dispensed at pharmacies during October to December 2005 and October to December 2006 were analysed, using adherence to the regional DTC guidelines as the main outcome measure. Adherence was assessed using the drug utilisation 90% methodology, i.e. focusing on drugs constituting 90% of the prescribed volume and the proportion of drugs included in the guidelines. The qualitative analysis focused on reports on the quality of drug prescribing submitted by each PHC in early 2007. The 139 PHCs participating in the programme accounted for 85% of all prescriptions issued in primary care during October to December 2006. Mean adherence to guidelines increased among participating practices by 3.3 percentage units (95% confidence interval (CI) 2.9-3.7%) to 83% (82.6-83.7%) during the year. The adherence among practices not participating increased by 3.1 percentage units (95% CI 1.7-4.4%) to 78.8% (95% CI 76.7-80.9%). The higher adherence achieved during the year corresponded to savings estimated at five times greater than the cost of running the programme including the financial incentives. In addition, many areas for improving prescribing were identified, such as limiting the prescribing of drugs with uncertain safety profiles and documentation as well as reporting adverse drug reactions. Although no causal effect can be attributed without a control group, we have shown the feasibility of a model linking payment to DTC adherence. This approach with its own quality assessment and goal setting offers an example to other regions and countries of how to increase the quality and efficiency of drug prescribing within limited resources.


Defining the T cell antigen proteome of wasp venom.
While modulation of T cell function is believed to be important in the successful acquisition of clinical tolerance during venom immunotherapy, little is known of the role of wasp venom specific T cell antigens. We sought comprehensively to characterize the T cell proteome for wasp venom to facilitate the future development of T cell-based immunotherapeutic approaches. Using peripheral blood mononuclear cells from wasp venom-allergic individuals and IL-4 ELISPOT analysis, we characterized T cell responses to whole venom and gel filtration/ion exchange-fractionated venom. Reactive fractions were purified and identified using highly sensitive electrospray ion-trap mass spectrometry. Wasp venom-allergic individuals have detectable whole wasp venom-specific T cells directly ex vivo, which show rapid IL-4 effector function. T cell responses to gel filtration/ion exchange fractionated venom were dominated by responses to phospholipase A(1), hyaluronidase and antigen 5. Although it is likely that there are many T cell antigens within wasp venom, the main responses are to proteins coincident with the known IgE-binding proteins.