Ethnicity and diagnosis in patients with affective disorders.
Clinically, African American psychiatric patients are disproportionately diagnosed with schizophrenia compared with white patients. Why this occurs is unknown. Extending prior work, the authors hypothesized that first-rank symptoms distract clinicians so that they fail to identify affective disorders in African Americans. 195 African American and white patients with at least 1 psychotic symptom (delusions, hallucinations, or prominent thought disorder) at admission were recruited from January 1, 1998, through May 31, 2001. Each patient received 3 independent DSM-IV diagnoses: a clinical diagnosis, a structured-interview diagnosis, and an expert-consensus diagnosis. The expert-consensus diagnoses were derived from the structured interviews, which were audiotaped and transcribed, and medical records. After reviewing edited transcripts and medical records from which ethnic cues had been eliminated, 2 psychiatrists assigned expert-consensus diagnoses and first-rank symptom ratings. For the 79 patients who received an expert-consensus diagnosis of an affective disorder, clinical variables, diagnoses, and first-rank symptoms were compared between African American (N = 39) and white (N = 40) patients. Seventy-nine (41%) of 195 patients were diagnosed with an affective disorder by expert consensus. African American men with an expert-consensus affective disorder were significantly (p <.03) more likely than other patients to be diagnosed with a schizophrenia spectrum disorder by clinical assessment and structured interview. Although first-rank symptoms were more commonly identified in African American men, this finding did not explain the difference in diagnoses. Post hoc analyses suggested that African American men diagnosed with a schizophrenia spectrum disorder were more likely than other patients to have been identified during structured interview as having psychotic symptoms in the absence of affective symptoms. The apparent misdiagnosis of schizophrenia in African-Americans with mood disorders cannot be ascribed to differences in first-rank symptoms. However, it may be due to a perception that psychotic symptoms are more chronic or persistent than affective symptoms in these patients.


Resveratrol protects against experimental stroke: putative neuroprotective role of heme oxygenase 1.
Epidemiological and experimental reports have linked mild-to-moderate wine and/or grape consumption to a lowered incidence of cardiovascular, cerebrovascular, and peripheral vascular risk. This study revealed that resveratrol, an enriched bioactive polyphenol in red wine, selectively induces heme oxygenase 1 (HO1) in a dose- and time-dependent manner in cultured mouse cortical neuronal cells and provides neuroprotection from free-radical or excitotoxicity damage. This protection was lost when cells were treated with a protein synthesis or heme oxygenase inhibitor, suggesting that HO1 induction is at least partially required for resveratrol's prophylactic properties. Furthermore, resveratrol pretreatment dose-dependently protected mice subjected to an optimized ischemic-reperfusion stroke model. Mice in which HO1 was selectively deleted lost most, if not all, of the beneficial effects. Together, the data suggest a potential intracellular pathway by which resveratrol can provide cell/organ resistance against neuropathological conditions.


Hypopituitarism after Gamma Knife surgery for postoperative nonfunctioning pituitary adenoma.
OBJECTIVEThis study investigated long-term follow-up data on the combined pituitary function test (CPFT) in patients who had undergone transsphenoidal surgery (TSS) for nonfunctioning pituitary adenoma (NFPA) to determine the clinical parameters indicative of hypopituitarism following postoperative Gamma Knife surgery (GKS).METHODSBetween 2001 and 2015, a total of 971 NFPA patients underwent TSS, and 76 of them (7.8%) underwent postoperative GKS. All 76 patients were evaluated with a CPFT before and after GKS. The hormonal states were analyzed based on the following parameters: relevant factors before GKS (age, sex, extent of resection, pre-GKS hormonal states, time interval between TSS and GKS), GKS-related factors (tumor volume; radiation dose to tumor, pituitary stalk, and normal gland; distance between tumor and stalk), and clinical outcomes (tumor control rate, changes in hormonal states, need for hormone-related medication due to hormonal changes).RESULTSOf the 971 NFPA patients, 797 had gross-total resection (GTR) and 174 had subtotal resection (STR). Twenty-five GTR patients (3.1%) and 51 STR patients (29.3%) underwent GKS. The average follow-up period after GKS was 53.5 ± 35.5 months, and the tumor control rate was 96%. Of the 76 patients who underwent GKS, 23 were excluded due to pre-GKS panhypopituitarism (22) or loss to follow-up (1). Hypopituitarism developed in 13 (24.5%) of the remaining 53 patients after GKS. A higher incidence of post-GKS hypopituitarism occurred in the patients with normal pre-GKS hormonal states (41.7%, 10/24) than in the patients with abnormal pre-GKS hormonal states (10.3%, 3/29; p = 0.024). Target tumor volume (4.7 ± 3.9 cm3), distance between tumor and pituitary stalk (2.0 ± 2.2 mm), stalk dose (cutoffs: mean dose 7.56 Gy, maximal dose 12.3 Gy), and normal gland dose (cutoffs: maximal dose 13.9 Gy, minimal dose 5.25 Gy) were factors predictive of post-GKS hypopituitarism (p < 0.05).CONCLUSIONSThis study analyzed the long-term follow-up CPFT data on hormonal changes in NFPA patients who underwent GKS after TSS. The authors propose a cutoff value for the radiation dose to the pituitary stalk and normal gland for the prevention of post-GKS hypopituitarism.


A systematic evaluation of multi-gene predictors for the pathological response of breast cancer patients to chemotherapy.
Previous studies have reported conflicting assessments of the ability of cell line-derived multi-gene predictors (MGPs) to forecast patient clinical outcomes in cancer patients, thereby warranting an investigation into their suitability for this task. Here, 42 breast cancer cell lines were evaluated by chemoresponse tests after treatment with either TFAC or FEC, two widely used standard combination chemotherapies for breast cancer. We used two different training cell line sets and two independent prediction methods, superPC and COXEN, to develop cell line-based MGPs, which were then validated in five patient cohorts treated with these chemotherapies. This evaluation yielded high prediction performances by these MGPs, regardless of the training set, chemotherapy, or prediction method. The MGPs were also able to predict patient clinical outcomes for the subgroup of estrogen receptor (ER)-negative patients, which has proven difficult in the past. These results demonstrated a potential of using an in vitro-based chemoresponse data as a model system in creating MGPs for stratifying patients' therapeutic responses. Clinical utility and applications of these MGPs will need to be carefully examined with relevant clinical outcome measurements and constraints in practical use.


Spatio-temporal patterns of genome evolution in allotetraploid species of the genus Oryza.
Despite knowledge that polyploidy is widespread and a major evolutionary force in flowering plant diversification, detailed comparative molecular studies on polyploidy have been confined to only a few species and families. The genus Oryza is composed of 23 species that are classified into ten distinct 'genome types' (six diploid and four polyploid), and is emerging as a powerful new model system to study polyploidy. Here we report the identification, sequence and comprehensive comparative annotation of eight homoeologous genomes from a single orthologous region (Adh1-Adh2) from four allopolyploid species representing each of the known Oryza genome types (BC, CD, HJ and KL). Detailed comparative phylogenomic analyses of these regions within and across species and ploidy levels provided several insights into the spatio-temporal dynamics of genome organization and evolution of this region in 'natural' polyploids of Oryza. The major findings of this study are that: (i) homoeologous genomic regions within the same nucleus experience both independent and parallel evolution, (ii) differential lineage-specific selection pressures do not occur between polyploids and their diploid progenitors, (iii) there have been no dramatic structural changes relative to the diploid ancestors, (iv) a variation in the molecular evolutionary rate exists between the two genomes in the BC complex species even though the BC and CD polyploid species appear to have arisen <2 million years ago, and (v) there are no clear distinctions in the patterns of genome evolution in the diploid versus polyploid species.


Detection of multidrug-resistant Salmonella enterica serovar typhimurium phage types DT102, DT104, and U302 by multiplex PCR.
Salmonella enterica serovar Typhimurium is a common cause of nontyphoidal salmonellosis in humans and animals. Multidrug-resistant serovar Typhimurium phage type DT104, which emerged in the 1990s, has become widely distributed in many countries. A total of 104 clinical isolates of Salmonella serogroup B were collected from three major hospitals in Taiwan during 1997 to 2003 and were examined by a multiplex PCR targeting the resistance genes and the spv gene of the virulence plasmid. A total of 51 isolates (49%) were resistant to all drugs (ACSSuT [resistance to ampicillin, chloramphenicol, streptomycin, sulfonamide, and tetracycline]), and all contained a 1.25-kb PCR fragment of integron that is part of the 43-kb Salmonella genomic island 1 (SGI1). The second group was resistant to SSu (28%), and the third was susceptible to all five drugs (13%). Fifty-nine isolates were serotyped to be serovar Typhimurium by the tube agglutination method using H antisera. The virulence plasmid was found in 54 (91.5%) of the 59 serovar Typhimurium isolates. A majority (94.1%) of the Salmonella serogroup B isolates with the ACSSuT resistance pattern harbored a virulence plasmid. Phage typing identified three major phage types: DT104, DT120, and U302. Analysis of the isolates by pulsed-field gel electrophoresis showed six genotypes. We found two genotypes in DT104 strains, two in DT120, and the other two in U302. The presence of a monophasic serovar (4,5,12:i:-) has added difficulty in the determination of the serovars of multidrug-resistant Salmonella serogroup B isolates. Nevertheless, the multiplex PCR devised in the present study appears to be efficient and useful in the rapid identification of ACSSuT-type serovar Typhimurium with SGI1, irrespective of their phage types.


Effectiveness of a School-Based Physical Activity Intervention on Cognitive Performance in Danish Adolescents: LCoMotion-Learning, Cognition and Motion - A Cluster Randomized Controlled Trial.
Physical activity is associated not only with health-related parameters, but also with cognitive and academic performance. However, no large scale school-based physical activity interventions have investigated effects on cognitive performance in adolescents. The aim of this study was to describe the effectiveness of a school-based physical activity intervention in enhancing cognitive performance in 12-14 years old adolescents. A 20 week cluster randomized controlled trial was conducted including seven intervention and seven control schools. A total of 632 students (mean (SD) age: 12.9 (0.6) years) completed the trial with baseline and follow-up data on primary or secondary outcomes (74% of randomized subjects). The intervention targeted physical activity during academic subjects, recess, school transportation and leisure-time. Cognitive performance was assessed using an executive functions test of inhibition (flanker task) with the primary outcomes being accuracy and reaction time on congruent and incongruent trials. Secondary outcomes included mathematics performance, physical activity levels, body-mass index, waist-circumference and cardiorespiratory fitness. No significant difference in change, comparing the intervention group to the control group, was observed on the primary outcomes (p's>0.05) or mathematics skills (p>0.05). An intervention effect was found for cardiorespiratory fitness in girls (21 meters (95% CI: 4.4-38.6) and body-mass index in boys (-0.22 kg/m2 (95% CI: -0.39-0.05). Contrary to our predictions, a significantly larger change in interference control for reaction time was found in favor of the control group (5.0 milliseconds (95% CI: 0-9). Baseline to mid-intervention changes in physical activity levels did not differ significantly between groups (all p's>0.05). No evidence was found for effectiveness of a 20-week multi-faceted school-based physical activity intervention for enhancing executive functioning or mathematics skills compared to a control group, but low implementation fidelity precludes interpretation of the causal relationship. www.ClinicalTrials.gov NCT02012881.


Cytokeratin 20-positive hepatocellular carcinoma.
The differential diagnosis between hepatocellular carcinoma (HCC), cholangiocarcinoma (CC) and metastatic colorectal adenocarcinoma (MCA) may be difficult when only based on morphology. For this purpose immunohistochemical analyses are often required, utilizing antibodies directed against CK8-18, Hep-Par1, glypican 3, CK7, CK19, CK20. Here we report a case of a 65-year-old man who presented with a clinical picture of decompensated cirrhosis. Ultrasonography revealed two nodular areas in the right liver lobe. Liver needle biopsy revealed micro-macronodular cirrhosis associated with HCC with trabecular and pseudoglandular patterns. Immunohistochemically, tumour cells were diffusely positive for CK8-18 and also diffusely immunostained by glypican 3 and Hep-Par1. Interestingly, a diffuse and strong staining for CK20 was detected in the vast majority of tumor cells, particularly in the areas showing a pseudo-glandular pattern. No immunostaining for CK7 and CK19 was found in the tumor cells. The tumor behaved aggressively, with a rapid diffusion to the whole liver. The patient died from the disease few months after presentation. These findings underline that the interpretation of the expression of CK20 alone in the differential diagnosis among HCC, CC and MCA should be done with caution because a diffuse immunoreactivity for CK20 alone may not rule out the diagnosis of HCC.


Vascular pseudoaneurysms in urology: clinical characteristics and management.
Vascular pseudoaneurysm is a well-documented complication that results after injury to an arterial wall. We review the urologic literature for all reported postprocedural and post-traumatic cases of pseudoaneurysm. A comprehensive review of the peer-reviewed literature was performed for reported cases of vascular pseudoaneurysm, with emphasis on clinical presentation and treatment options. Vascular pseudoaneurysm is a high-pressure collection of blood that communicates directly with an arterial vessel. The clinical presentation varies, depending on the location and size of the lesion, and can be quite dramatic and potentially life threatening. There may be an increasing incidence of pseudoaneurysms after minimally invasive and laparoscopic techniques. Selective and superselective embolization is the preferred treatment for patients with vascular pseudoaneurysms. Vascular pseudoaneurysm is an important but rare complication that is increasingly reported after minimally invasive urologic surgery. This diagnosis necessitates a high index of suspicion and radiologic acumen. Treatment is individually tailored to each patient, but selective embolization appears to be the standard of care.


Intermittent Cerebrospinal Leak After Inadvertent Dural Puncture During Epidural Catheter Placement for Postoperative Analgesia.
Regional anesthesia is being used more frequently in pediatric anesthesia practice, including the perioperative care of neonates and infants. Adverse effects may be encountered during epidural needle placement, with catheter advancement, or subsequently during infusion of local anesthetic agents. We present the rare occurrence of a persistent cerebrospinal fluid (CSF) leak following inadvertent dural puncture (wet tap) during attempted placement of an epidural catheter in a 6-year-old child. Potential adverse effects of epidural anesthesia in children are discussed, and options for treatment of a persistent CSF leak after inadvertent dural puncture are reviewed.


Fibrinogen alpha chain promotes the migration and invasion of human endometrial stromal cells in endometriosis through focal adhesion kinase/protein kinase B/matrix metallopeptidase 2 pathway†.
Fibrinogen alpha chain (FGA), a cell adhesion molecule, contains two arginyl-glycyl-aspartic acid (RGD) cell adhesion sequences. Our previous study demonstrated that FGA, as an up-regulated protein in endometriosis (EM), was closely related to disease severity and involved in the development of EM. However, the biological functions and underlying mechanism of FGA in EM have not been fully understood. To explore the roles of FGA in EM, we analyzed the effects of FGA on the biological behaviors of human primary eutopic endometrial stromal cells (EuESC). The results indicated FGA knockdown suppressed the migration and invasion ability of EuESC, which also altered the distribution of cytoskeletal filamentous and cell morphology. Western blot analysis demonstrated that knockdown of FGA attenuated the migration-related protein levels of vimentin and matrix metallopeptidase 2 (MMP-2), but not integrin subunit alpha V (ITGAV) and integrin subunit beta 3 (ITGB3). Meanwhile, integrin-linked transduction pathways were detected. We found FGA knockdown significantly suppressed the expression of focal adhesion kinase (FAK) level and protein kinase B (AKT) phosphorylation, without extracellular-signal-regulated kinase (ERK) dependent pathways. Treatment with the AKT inhibitor MK2206 or RGD antagonist highly decreased the effects of FGA on the migration and invasion of EuESC. RGD antagonist treatment strongly inhibited FAK- and AKT-dependent pathways, but not ERK pathways. Our data indicated that FGA may enhance the migration and invasion of EuESC through RGD sequences binding integrin and activating the FAK/AKT/MMP-2 signaling pathway. This novel finding suggests that FGA may provide a novel potential approach to the treatment of EM, which provides a new way to understand the pathogenesis of EM.


Characterization of two populations of ectopic cells isolated from the hearts of NGF transgenic mice.
We have observed sympathetic hyperinnervation and hyperplasia of an unknown cell population within the base of the hearts from transgenic mice in which nerve growth factor (NGF) expression was targeted to cardiac tissue (Hassankhani et al., 1993, 1995). To characterize this ectopic cell population, cells from the base of hearts of 3- to 4-week-old transgenic mice were dissociated and established in tissue culture, and their properties were analyzed using morphological, immunocytochemical, and physiological techniques. Morphological studies of the ectopic cells revealed the temporal expression of two different cellular phenotypes: (i) a spindle-shaped cell type, present by 1 to 2 days in vitro (DIV) and (ii) cells with a fiber-like morphology, detected by 7 DIV. In contrast to transgenic cardiac myocytes that express immunoreactivity to antibodies against sarcomeric myosin, beat spontaneously (approximately 60/min), and are electrically excitable, the ectopic cell types did not stain with an antibody against sarcomeric myosin, never showed contractile activity, and did not manifest membrane excitability. Moreover, these two types of ectopic cells demonstrated other distinctive characteristics. The spindle-shaped subpopulation typically formed small clusters of cells that were immunostained by antibodies to GFAP, vimentin, and low-affinity NGF receptor (LNGFR) and demonstrated a low incidence of dye coupling (30%), whereas the fiber-like cells aligned themselves along their long axes, immunostained for gp140trkA, LNGFR, and anti alpha-smooth muscle actin and showed extensive (100%) and diffuse dye spread to numerous other contiguous fiber-like cells. Dye spread was not observed between the adjacent fiber-like and the spindle-shaped cells and was reversibly blocked between homologous cells after treatment with halothane, a gap junction channel blocking agent. In comparison to cardiac myocytes, macroscopic junctional conductance of the spindle-shaped cells was more voltage-sensitive. At the single channel level, unitary junctional conductances of approximately 60 pS were predominant. Overall, these results indicate that these ectopic cells are likely derived from neural crest. The spindle-shaped cells appear to be immature Schwann cells, whereas the fiber-like cells may be related to the ectomesenchymal cells contributing to the morphogenesis of the cardiac outflow tract.


Survival times for cats with hyperthyroidism treated with iodine 131, methimazole, or both: 167 cases (1996-2003).
To compare survival times for cats with hyperthyroidism treated with iodine 131, methimazole, or both and identify factors associated with survival time. Retrospective case series. 167 cats. Medical records of cats in which hyperthyroidism had been confirmed on the basis of high serum thyroxine concentration, results of thyroid scintigraphy, or both were reviewed. 55 (33%) cats were treated with 131I alone, 65 (39%) were treated with methimazole followed by 131I, and 47 (28%) were treated with methimazole alone. Twenty-four of 166 (14%) cats had preexisting renal disease, and 115 (69%) had preexisting hepatic disease. Age was positively correlated (r = 0.4) with survival time, with older cats more likely to live longer. Cats with preexisting renal disease had significantly shorter survival times than did cats without preexisting renal disease. When cats with preexisting renal disease were excluded, median survival time for cats treated with methimazole alone (2.0 years; interquartile range [IQR], 1 to 3.9 years) was significantly shorter than median survival time for cats treated with 131I alone (4.0 years; IQR, 3.0 to 4.8 years) or methimazole followed by 131I (5.3 years; IQR, 2.2 to 6.5 years). Results suggest that age, preexisting renal disease, and treatment type were associated with survival time in cats undergoing medical treatment of hyperthyroidism.


Caring for Africa's sickle cell children: will we rise to the challenge?
Most of the world's sickle cell disease (SCD) burden is in Africa, where it is a major contributor to child morbidity and mortality. Despite the low cost of many preventive SCD interventions, insufficient resources have been allocated, and progress in alleviating the SCD burden has lagged behind other public-health efforts in Africa. The recent announcement of massive new funding for research into curative SCD therapies is encouraging in the long term, but over the next few decades, it is unlikely to help Africa's SCD children substantially. A major barrier to progress has been the absence of large-scale early-life screening. Most SCD deaths in Africa probably occur before cases are even diagnosed. In the last few years, novel inexpensive SCD point-of-care test kits have become widely available and have been deployed successfully in African field settings. These kits could potentially enable universal early SCD screening. Other recent developments are the expansion of the pneumococcal conjugate vaccine towards near-universal coverage, and the demonstrated safety, efficacy, and increasing availability and affordability of hydroxyurea across the continent. Most elements of standard healthcare for SCD children that are already proven to work in the West, could and should now be implemented at scale in Africa. National and continental SCD research and care networks in Africa have also made substantial progress, assembling care guidelines and enabling the deployment and scale-up of SCD public-health systems. Substantial logistical, cultural, and awareness barriers remain, but with sufficient financial and political will, similar barriers have already been overcome in efforts to control other diseases in Africa. Despite remaining challenges, several high-SCD-burden African countries have the political will and infrastructure for the rapid implementation and scale-up of comprehensive SCD childcare programs. A globally funded effort starting with these countries and expanding elsewhere in Africa and to other high-burden countries, including India, could transform the lives of SCD children worldwide and help countries to attain their Sustainable Development Goals. This endeavor would also require ongoing research focused on the unique needs and challenges of SCD patients, and children in particular, in regions of high prevalence.


Effect of phorbol esters on iron uptake in human hematopoietic cell lines.
We have investigated the effect of 12-O-tetradecanoylphorbol-13-acetate (TPA) on iron uptake into human hematopoietic cell lines K562, U937, and HL-60. TPA inhibited both cell growth and iron uptake by these cell lines. This effect was rapid, which is typical of phorbol esters which are biologically active, and it occurred at very low concentrations of TPA. This effect of TPA was dependent upon an inhibition of the transferrin-binding capacity as estimated on intact cells. However, experiments with transferrin binding on cell samples dissolved in 1% Triton X-100 showed that TPA-treated cells exhibited a transferrin-binding capacity similar to that of control cells. On the basis of this result, it is suggested that TPA modified a part of transferrin receptors present in the cells; as a result of this modification, these receptors became unavailable for binding transferrin, but they remained physically present in the cell. Other compounds capable of inducing the differentiation of leukemic cells, such as dimethyl sulfoxide, butyrate, retinoic acid, and 1 alpha,25-dihydroxy-vitamin D3, did not acutely inhibit iron uptake. We also investigated the effect of TPA on transferrin receptors in a cellular system in which phorbol esters stimulate cell proliferation. At 16 X 10(-9) M, TPA markedly stimulated the proliferation of T-lymphocytes. However, in spite of this marked stimulation of cell proliferation, TPA-stimulated lymphocytes exhibited a transferrin-binding capacity much inferior to cells stimulated by other mitogens, such as phytohemagglutinin.


Endogenous Stochastic Decoding of the CUG Codon by Competing Ser- and Leu-tRNAs in Ascoidea asiatica.
Although the "universal" genetic code is now known not to be universal, and stop codons can have multiple meanings, one regularity remains, namely that for a given sense codon there is a unique translation. Examining CUG usage in yeasts that have transferred CUG away from leucine, we here report the first example of dual coding: Ascoidea asiatica stochastically encodes CUG as both serine and leucine in approximately equal proportions. This is deleterious, as evidenced by CUG codons being rare, never at conserved serine or leucine residues, and predominantly in lowly expressed genes. Related yeasts solve the problem by loss of function of one of the two tRNAs. This dual coding is consistent with the tRNA-loss-driven codon reassignment hypothesis, and provides a unique example of a proteome that cannot be deterministically predicted. VIDEO ABSTRACT.


Compatibility between livestock databases used for quantitative biosecurity response in New Zealand.
To characterise New Zealand's livestock biosecurity databases, and investigate their compatibility and capacity to provide a single integrated data source for quantitative outbreak analysis. Contemporary snapshots of the data in three national livestock biosecurity databases, AgriBase, FarmsOnLine (FOL) and the National Animal Identification and Tracing Scheme (NAIT), were obtained on 16 September, 1 September and 30 April 2014, respectively, and loaded into a relational database. A frequency table of animal numbers per farm was calculated for the AgriBase and FOL datasets. A two dimensional kernel density estimate was calculated for farms reporting the presence of cattle, pigs, deer, and small ruminants in each database and the ratio of farm densities for AgriBase versus FOL calculated. The extent to which records in the three databases could be matched and linked was quantified, and the level of agreement amongst them for the presence of different species on properties assessed using Cohen's kappa statistic. AgriBase contained fewer records than FOL, but recorded animal numbers present on each farm, whereas FOL contained more records, but captured only presence/absence of animals. The ratio of farm densities in AgriBase relative to FOL for pigs and deer was reasonably homogeneous across New Zealand, with AgriBase having a farm density approximately 80% of FOL. For cattle and small ruminants, there was considerable heterogeneity, with AgriBase showing a density of cattle farms in the Central Otago region that was 20% of FOL, and a density of small ruminant farms in the central West Coast area that was twice that of FOL. Only 37% of records in FOL could be linked to AgriBase, but the level of agreement for the presence of different species between these databases was substantial (kappa>0.6). Both NAIT and FOL shared common farm identifiers which could be used to georeference animal movements, and there was a fair to substantial agreement (kappa 0.32-0.69) between these databases for the presence of cattle and deer on properties. The three databases broadly agreed with each other, but important differences existed in both species composition and spatial coverage which raises concern over their accuracy. Importantly, they cannot be reliably linked together to provide a single picture of New Zealand's livestock industry, limiting the ability to use advanced quantitative techniques to provide effective decision support during disease outbreaks. We recommend that a single integrated database be developed, with alignment of resources and legislation for its upkeep.


Turbidity, birefringence, and fluorescence changes in skeletal muscle coincident with the action potential.
Electrical stimulation of frog striated muscle was found to prduce trasient changes in turbidity. birefringence, and fluorescence (of fibers stained with pyronine B). The initial phase of these optical changes Was coinicident with the action potential. These findings suggest that there is a macromolecular conformational change in muscle membrane during excitation.


B7-DC-Ig enhances vaccine effect by a novel mechanism dependent on PD-1 expression level on T cell subsets.
Programmed death receptor 1 (PD-1) is an important signaling molecule often involved in tumor-mediated suppression of activated immune cells. Binding of this receptor to its ligands, B7-H1 (PD-L1) and B7-DC (PD-L2), attenuates T cell activation, reduces IL-2 and IFN-γ secretion, decreases proliferation and cytotoxicity, and induces apoptosis. B7-DC-Ig is a recombinant protein that binds and targets PD-1. It is composed of an extracellular domain of murine B7-DC fused to the Fc portion of murine IgG2a. In this study, we demonstrate that B7-DC-Ig can enhance the therapeutic efficacy of vaccine when combined with cyclophosphamide. We show that this combination significantly enhances Ag-specific immune responses and leads to complete eradication of established tumors in 60% of mice and that this effect is CD8 dependent. We identified a novel mechanism by which B7-DC-Ig exerts its therapeutic effect that is distinctly different from direct blocking of the PD-L1-PD-1 interaction. In this study, we demonstrate that there are significant differences between levels and timing of surface PD-1 expression on different T cell subsets. We found that these differences play critical roles in anti-tumor immune effect exhibited by B7-DC-Ig through inhibiting proliferation of PD-1(high) CD4 T cells, leading to a significant decrease in the level of these cells, which are enriched for regulatory T cells, within the tumor. In addition, it also leads to a decrease in PD-1(high) CD8 T cells, tipping the balance toward nonexhausted functional PD-1(low) CD8 T cells. We believe that the PD-1 expression level on T cells is a crucial factor that needs to be considered when designing PD-1-targeting immune therapies.


Recruitment and Retention of Community Preceptors.
Recruiting and retaining community-based pediatricians for teaching medical students has been explored through the lens of preceptors and educational leaders. The purpose of this study was to explore the perspective of pediatric department chairs, a key stakeholder group charged with maintaining teaching capacity among a faculty. In 2015, members of the Association of Medical School Pediatric Department Chairs and Council on Medical Student Education in Pediatrics joint task force disseminated a 20-item survey to pediatric department chairs in the United States and Canada. Topics included demographics, incentives offered to community pediatricians, and the perceived value and feasibility of such incentives. Data were analyzed using descriptive statistics and χ2 to compare categorical variables. Pediatric department chairs from 92 of 145 (63% response rate) medical schools returned the survey. Sixty-seven percent reported difficulty recruiting or retaining preceptors, and 51% reported high-reliance on preceptors for the ambulatory portion of the pediatrics clerkship. Almost all (92%) cited competition from other programs for the services of community preceptors. The provision of incentives was correlated with perceived feasibility (R2 = 0.65) but not their perceived value (R2 = 0.12). Few (21%) chairs reported providing financial compensation to preceptors. The provision of compensation was not related to reliance but did vary significantly by geographical region (P < .001). Pediatric departments rely heavily on community-based pediatricians but face competition from internal and external training programs. The perspective of department chairs is valuable in weighing interventions to facilitate continued recruitment and retention of community preceptors.


Comparing transcription rate and mRNA abundance as parameters for biochemical pathway and network analysis.
The cells adapt to extra- and intra-cellular signals by dynamic orchestration of activities of pathways in the biochemical networks. Dynamic control of the gene expression process represents a major mechanism for pathway activity regulation. Gene expression has thus been routinely measured, most frequently at steady-state mRNA abundance level using micro-array technology. The results are widely used in statistical inference of the structures of underlying biochemical networks, with the assumption that functionally related genes exhibit similar dynamic profiles. Steady-state mRNA abundance, however, is a composite of two factors: transcription rate and mRNA degradation rate. The question being asked here is therefore whether steady-state mRNA abundance or any of two factors is a more informative measurement target for studying network dynamics. The yeast S. cerevisiae was used as model organism and transcription rate was chosen out of the two factors in this study, because genome-wide determination of transcription rates has been reported for several physiological processes in this species. Our strategy is to test which one is a better measurement of functional relatedness between genes. The analysis was performed on those S. cerevisiae genes that have bacterial orthologs as identified by reciprocal BLAST analysis, so that functional relatedness of a gene pair can be measured by the frequency at which their bacterial orthologs co-occur in the same operon in the collection of bacterial genomes. It is found that transcription rate data is generally a better parameter for functional relatedness than steady state mRNA abundance, suggesting transcription rate data is more informative to use in deciphering the logics used by the cells in dynamic regulation of biochemical network behaviors. The significance of this finding for network and systems biology, as well as biomedical research in general, is discussed.


Heart and Neural Crest Derivatives Expressed Transcript 2 (HAND2): a novel biomarker for the identification of atypical hyperplasia and Type I endometrial carcinoma.
Progesterone inhibits the proliferative growth effects of estrogen in the endometrium and prevents the development of endometrial hyperplasia and Type I adenocarcinoma. The exact mechanism of this action is unknown. The progesterone-induced helix-loop-helix transcription factor Heart and Neural Crest Derivatives Expressed 2 (Hand2) was recently shown to suppress production of growth factors in the endometrium. In Hand2 knockout mice, continuous proliferation of the endometrium was observed. In this study, archival paraffin-embedded tissue from 56 hysterectomy specimens was examined by immunohistochemistry for the expression and localization of Hand2, estrogen receptor (ER), progesterone receptor (PR), and Ki-67. Diagnoses included disordered proliferative endometrium, simple and complex hyperplasia with or without atypia, and endometrioid adenocarcinoma. Hand2 expression is localized to endometrial stromal nuclei. In benign endometrium, Hand2 expression was moderate to strong (10/11; 91%), with weak Hand2 expression in only 1 case (1/11; 9%). Similar Hand2 expression patterns were observed in disordered proliferative endometrium and simple hyperplasia without atypia, with moderate to strong expression in 91% of cases (10/11) and weak expression in 9% of cases (1/11). In contrast, simple and complex hyperplasia with atypia exhibited moderate to strong Hand2 expression in 8% of cases (1/12) and a loss of expression or weak expression in 92% of cases (11/12). In endometrioid adenocarcinomas, Hand2 expression was absent in all cases (22/22). Hand2 is expressed in the stroma of benign endometrium, but expression is significantly reduced or lost in atypical hyperplasia and endometrioid carcinoma. Thus, the absence of Hand2 expression may be a useful biomarker for atypical hyperplasia and endometrioid carcinoma.


Management of trapeziometacarpal osteoarthritis pain and dysfunction using mobilization with movement technique in combination with kinesiology tape: a case report.
The purpose of this report is to describe the management of a patient with advanced trapeziometacarpal (TMC) osteoarthritis (OA) using mobilization with movement technique in combination with kinesiology tape to decrease pain and improve range of motion. A 52-year-old female seamstress (a career of 35 years' duration) presented to a physiotherapy clinic with pain in the dorsal aspect of the thumb carpometacarpal region of the right (dominant) hand. Examination revealed reduced ability to abduct the right thumb, significant loss of web space, weakness of pinch grip, and deterioration of hand function. Radiographs demonstrated OA of the TMC stage IV according to the Eaton-Littler-Burton classification, with instability and subluxation of the joint. A combined treatment protocol of mobilization with movement and kinesiology tape at the TMC joint for 12 weekly sessions was performed. Outcome measures were assessed at baseline, immediately upon completion of treatment, and at 2-month follow-up and included numeric pain rating scale, range of motion, pressure pain threshold, and tip pinch strength at the TMC joint. Treatment interventions were applied for 12 sessions over a period of 2 months. Outcome measures indicated significant reduction of the patient's subjective pain reports and considerable improvement in functional and occupational tasks. A follow-up visit at 4 months (2 months after last treatment) showed that the improvement was maintained. A combined program of mobilization with movement and kinesiology tape reduced pain, increased range of motion, and increased tip pinch strength in a patient with severe functional impairment related to dominant TMC OA.


Complications and Unfavorable Clinical Outcomes in Obese and Overweight Patients Treated for Adult Lumbar or Thoracolumbar Scoliosis With Combined Anterior/Posterior Surgery.
Combined anterior/posterior (A/P) spinal fusion with instrumentation has been used for many years in the treatment of adult thoracolumbar and lumbar (TL/L) scoliosis. However, the risk factors for complications and poor clinical outcomes with this procedure are not well known. To assess the risk factors for poor clinical outcomes in a series of adult lumbar or scoliosis patients undergoing combined A/P-instrumented spinal fusion. This study was a retrospective case series of surgically treated adult lumbar or thoracolumbar scoliosis patients. From 1998 to 2006, 57 patients with diagnoses of adult idiopathic scoliosis or degenerative TL/L scoliosis underwent combined A/P spinal instrumentation and fusion at 1 institution, performed by 1 senior author. The preoperative and postoperative outcome measurements included self-report measurements, physiological measurements, and functional measurements. A retrospective review of this patient group was performed to evaluate patient satisfaction, functional outcomes, pain, curve progression, and complications. Radiographic measurements included coronal balance, sagittal vertical axis, Cobb angle, thoracic kyphosis, lumbar lordosis, and pelvic incidence preoperatively, immediately postoperatively, and during follow-up. In terms of risk factors, bone mineral density, body mass index, age, kyphosis, and fusion to the sacrum were reviewed. Postoperative Scoliosis Research Society Patient Questionnaire outcome scores, Oswestry Disability Index (ODI), and anterior surgical site pain (ASSP) were also evaluated. Means were compared with the Student t test and the χ test. Logistic regression analyses were used to predict the probabilities and the odds ratios (ORs) of the risk factors for poor clinical outcomes. A P-value of <0.05 with a confidence interval of 95% was considered significant. Fifty patients had adult idiopathic scoliosis, and 7 patients had degenerative scoliosis. The average age at surgery was 53.8 years (34-74 y), and the average follow-up was 4.8 years (2-11 y). Coronal correction for thoracic, thoracolumbar, and lumbosacral curves improved significantly. The degree of sagittal curve and coronal and sagittal balance were not significantly changed after surgery or at the final follow-up. ODI, the pain intensity domain of the ODI, and ASSP were significantly worse in obese and overweight patients, whereas OR time, estimated blood loss, and number of fused vertebrae were not different in the entire group (P=0.03 for ODI, P=0.002 for pain domain of ODI, and P=0.003 for ASSP). Logistic regression analyses for the risk factors of poor clinical outcomes indicated obesity and overweight as risk factors for poor clinical outcomes (OR=6.25 for ODI and 5.88 for ODI pain intensity score). A significantly higher rate of major complications occurred in this group compared to the entire group (30.4%, P=0.04). Low bone mineral density, old age, kyphosis, and fusion to the sacrum were not risk factors for poor clinical outcomes. Despite the good function scores and acceptable pain levels in most patients, the ODI scores of obese and overweight patients were worse compared to the rest of the patients in this study. Significantly worse scores on the pain intensity domain of the ODI and ASSP differences were likely caused by extensive dissection of the abdominal wall and psoas muscles and the technical difficulty of achieving an anterior approach to the thoracolumbar spine. Radiographs revealed no progression of the TL/L curves. This study indicates that obesity and overweight are potential risk factors for combined A/P-instrumented spinal fusion in patients with adult TL/L scoliosis, perhaps due to the technical difficulty of achieving an anterior approach to the thoracolumbar spine.


Recipient vessel selection in the difficult neck: Outcomes of external carotid artery transposition and end-to-end microvascular anastomosis.
Selection of recipient vessels for head and neck microvascular surgery may be limited in the previously dissected or irradiated neck. When distal branches of the external carotid artery (ECA) are unavailable, additional options for arterial inflow are needed. Here we propose high ligation of the ECA and transposition toward the lower neck as an alternative. After obtaining institutional approval, patients who underwent head and neck tumor resection and simultaneous free flap reconstruction were identified over a 5-year period. Patients whose recipient artery was listed in the operative report were included. Chart review was performed to identify patient demographics, operative details, and patient and flap complications. In cases where the ECA was used, the artery was traced distally with care taken to protect the hypoglossal nerve. The ECA was then divided and transposed toward the lower neck where an end-to-end microvascular anastomosis was performed. The recipient artery used for head and neck microsurgery was available for 176 flaps, and the facial (n = 127, 72.2%) and external carotid (n = 19, 10.8%) arteries were most commonly used. There were 0 flap thromboses in the ECA group compared to 3 flap thromboses that occurred with other recipient arteries (P = 1.00). No cases of first bite syndrome or hypoglossal nerve injury were identified. The ECA may be transposed toward the lower neck and used for end-to-end microvascular anastomosis without complication of hypoglossal nerve injury or first bite syndrome. This method may be considered an alternative in patients with limited recipient vessel options for head and neck microsurgery. © 2015 Wiley Periodicals, Inc. Microsurgery 37:96-100, 2017.


What prevents midwifery quality care in Bangladesh? A focus group enquiry with midwifery students.
With professional midwives being introduced in Bangladesh in 2013, the aim of this study was to describe midwifery students perceptions on midwives' realities in Bangladesh, based on their own experiences. Data were collected through 14 focus group discussions that included a total of 67 third-year diploma midwifery students at public nursing institutes/colleges in different parts of Bangladesh. Data were analyzed deductively using an analytical framework identifying social, professional and economical barriers to the provision of quality care by midwifery personnel. The social barriers preventing midwifery quality care falls outside the parameters of Bangladeshi cultural norms that have been shaped by beliefs associated with religion, society, and gender norms. This puts midwives in a vulnerable position due to cultural prejudice. Professional barriers include heavy workloads with a shortage of staff who were not utilized to their full capacity within the health system. The reason for this was a lack of recognition in the medical hierarchy, leaving midwives with low levels of autonomy. Economical barriers were reflected by lack of supplies and hospital beds, midwives earning only low and/or irregular salaries, a lack of opportunities for recreation, and personal insecurity related to lack of housing and transportation. Without adequate support for midwives, to strengthen their self-confidence through education and through continuous professional and economic development, little can be achieved in terms of improving quality care of women during the period around early and late pregnancy including childbirth.The findings can be used for discussions aimed to mobilize a midwifery workforce across the continuum of care to deliver quality reproductive health care services. No matter how much adequate support is provided to midwives, to strengthen their self-confidence through education, continuous professional and economic development, addressing the social barriers is a prerequisite for provision of quality care.


Angiogenesis and lymphangiogenesis in parathyroid proliferative lesions.
Angiogenesis and lymphangiogenesis are involved in tumoral growth and metastatic spread. There is little information on angiogenesis and no available data on lymphangiogenesis in parathyroid glands (PTG). Using immunohistochemistry for CD34, LYVE-1 (specific markers for vascular and lymphatic endothelium, respectively), vascular endothelial growth factor (VEGF)-A, VEGF-C, and fibroblast growth factor (FGF)-2, this study analyzes microvascular density (MVD), lymphatic vascular density (LVD), and expression of angiogenic and lymphangiogenic growth factors in 13 normal PTG, 77 parathyroid adenomas (PTA), and 17 primary parathyroid hyperplasia (PPH). MVD was higher in PPH and PTA, compared with PTG (P < 0.001). There was no difference in VEGF-A expression among groups. In contrast, FGF-2 expression was higher in PPH, compared with PTA and PTG (P < 0.0001). FGF-2 scores and MVD were significantly correlated (r = 0.43). LVD did not differ among groups, and VEGF-C expression was unrelated to LVD. There was no relationship between MVD and tumor behavior (adenoma size, PTH, or calcium). In conclusion, this study shows increased angiogenesis in parathyroid proliferative lesions compared with normal glands and suggests that FGF-2 is proangiogenic in parathyroid tissue. In PTA, tumor behavior is not related to angiogenic phenotype. This is the first demonstration of lymphatic vessels in PTG, but the lack of correlation with VEGF-C expression suggests that VEGF-C is not the primary lymphangiogenic factor.


Meshed Architecture of Performance as a Model of Situated Cognition.
In this paper, we engage in a reciprocal analysis of situated cognition and the notion of "meshed architecture" as found in performance studies (Christensen et al., 2016). We start with an account of various conceptions of situated cognition using the distinction between functional integration, which characterizes how an agent dynamically organizes to couple with its environment, and task dependency, which specifies various constraints and structures imposed by the environment (see Slors, 2019). We then exploit the concept of a meshed architecture as a model that provides a more focused analysis of situated cognition and performance. Through this analysis, we show how the model of meshed architecture can be enhanced through (1) the involvement of a more complex set of cognitive processes, (2) a form of intrinsic control, (3) the influence of affective factors, and (4) the role of factors external to the performer. The aim of this paper, then, is twofold: first to work out an enhanced conception of the model of meshed architecture by taking into consideration a number of factors that clarify its situated nature, and second, to use this model to provide a richer and more definitive understanding of the meaning of situated cognition. Thus, we argue that this reciprocal analysis gives us a very productive way to think about how various elements come together in skilled action and performance but also a detailed way to characterize situated cognition.


Rates of sensitivity reactions to aspirin: problems in interpreting the data.
This work was done to determine the reasons for variation in the reported rates--ranging from less than 1% to greater than 50%--of sensitivity to aspirin and cross-reactivity to acetaminophen and ibuprofen. In 47 studies that reported rates of sensitivity and in 23 reports that contained series of sensitive patients, we examined the research setting, source of patients, clinical attributes of the study group, admission process, and selection, operational definition, and method of determining sensitivity reactions. In five studies with reasonably well-specified methods, the reported sensitivity rates to aspirin were lowest (0.3% to 0.9%) for patients without allergic tendencies, higher in asthmatics, and highest if patients had nasal polyps or severe atopy. Although not determined in any of these studies, the rate of sensitivity in a general (nonclinical) population would doubtlessly be substantially lower than the rate of three per 1000 reported for nonallergic patients. The admixture of different clinical groups, varying definitions, and ascertainment of a sensitivity reaction seem to be responsible for the variations in the reported rates of sensitivity and cross-reactivity.


Interleukin-10 polymorphism in position -1082 and acute respiratory distress syndrome.
The GG genotype of the interleukin (IL)-10 promoter polymorphism in position -1082 (-1082GG) has been associated with increased IL-10 production. The current authors hypothesised that the -1082GG genotype is associated with the development of, and outcomes in, acute respiratory distress syndrome (ARDS). A nested case-control study was conducted in 211 Caucasian cases of ARDS and 429 controls who were admitted to an intensive care unit with sepsis, trauma, aspiration or massive transfusions. Cases were followed for organ failure and 60-day mortality. The -1082GG genotype was associated with the development of ARDS, but only in the presence of a significant interaction between the -1082GG genotype and age. Among patients with ARDS, the -1082GG genotype was associated with decreased severity of illness on admission, lower daily organ dysfunction scores and lower 60-day mortality. In conclusion, the high interleukin-10-producing -1082GG genotype may be associated with variable odds for acute respiratory distress syndrome development depending on age. Among those with acute respiratory distress syndrome, the -1082GG genotype is associated with lower mortality and organ failure. Further studies are needed to confirm these findings.


Ribo-Pop: simple, cost-effective, and widely applicable ribosomal RNA depletion.
The measurement of RNA abundance derived from massively parallel sequencing experiments is an essential technique. Methods that reduce ribosomal RNA levels are usually required prior to sequencing library construction because ribosomal RNA typically comprises the vast majority of a total RNA sample. For some experiments, ribosomal RNA depletion is favored over poly(A) selection because it offers a more inclusive representation of the transcriptome. However, methods to deplete ribosomal RNA are generally proprietary, complex, inefficient, applicable to only specific species, or compatible with only a narrow range of RNA input levels. Here, we describe Ribo-Pop (ribosomal RNA depletion for popular use), a simple workflow and antisense oligo design strategy that we demonstrate works over a wide input range and can be easily adapted to any organism with a sequenced genome. We provide a computational pipeline for probe selection, a streamlined 20-min protocol, and ready-to-use oligo sequences for several organisms. We anticipate that our simple and generalizable "open source" design strategy would enable virtually any laboratory to pursue full transcriptome sequencing in their organism of interest with minimal time and resource investment.


Characterization of multiple [3H]ryanodine binding sites on the Ca2+ release channel of sarcoplasmic reticulum from skeletal and cardiac muscle: evidence for a sequential mechanism in ryanodine action.
Kinetic and equilibrium measurements of [3H]ryanodine binding to the Ca2+ release channel of rabbit skeletal and rat cardiac sarcoplasmic reticulum (SR) are examined to ascertain the nature of cooperative interactions among high and low affinity binding sites and to quantitate their distribution. Equilibrium studies reveal affinities of 1-4 nM for the highest affinity binding site and of 30-50 nM, 500-800 nM, and 2-4 microM for the lower affinity sites in both preparations, with Hill coefficients of significantly less than 1, and initial rates of association and dissociation increase with increasing concentrations of ryanodine. SR vesicles are actively loaded in the presence of pyrophosphate, and fluctuations in extravesicular Ca2+ are measured by the absorbance change of antipyrylazo III. The data demonstrate a biphasic, time- and concentration-dependent action of ryanodine on the release of Ca2+, with an initial activation and a subsequent inactivation phase. Kinetic analysis of the activation of Ca2+ release by ryanodine, in consonance with the binding data, demonstrates the existence of multiple binding sites for the alkaloid on the channel complex, with nanomolar to micromolar affinities. Based on the present findings obtained by receptor binding analysis and Ca2+ transport measurements, we suggest a model that describes four, most plausibly negatively cooperative, binding sites on the Ca2+ release channel. Occupation of ryanodine binding sites produces sequential activation followed by inactivation of the SR channel, revealing the strong possibility of an irreversible uncoupling of the native function of the receptor/channel complex by high concentrations of ryanodine. A model relating ryanodine receptor occupancy with SR Ca2+ release stresses two important new findings regarding the interaction of ryanodine with its receptor. First, ryanodine binds to four sites on the oligomeric channel complex with decreasing affinities, which can be best described by allosteric negative cooperativity. Second, binding of ryanodine to its receptor activates the Ca2+ release channel in a concentration-dependent and saturable manner in the range of 20 nM to 1 mM and produces a kinetically limited and sequential inactivation of the Ca2+ channel, with the concomitant attainment of full negative cooperativity. The results presented suggest that driving of the complex toward full negative cooperativity with high concentrations of ryanodine promotes a long-lived conformational state in which ryanodine is physically occluded and hindered from free diffusion from its binding site.


ALK and crizotinib: after the honeymoon…what else? Resistance mechanisms and new therapies to overcome it.
The last few decades have witnessed a silent revolution in the war against NSCLC, thanks to the discovery of "oncogenic drivers" and the subsequent development of targeted therapies. The discovery of the EML4-ALK fusion gene in a subgroup of patients with NSCLC and the subsequent clinical development of crizotinib has been an amazing success story in lung cancer translational-research, and its accelerated approval [only 4 years from the discovery of ALK rearrangement in NSCLC to the approval by the Food and Drug Administration (FDA)] marked the beginning of the new decade of targeted therapy. However, common to all targeted therapies, despite an initial benefit, patients inevitably experience tumor progression, due to the development of resistance. Several molecular mechanisms are responsible for acquired resistance, such as secondary mutations of ALK kinase domain or amplification of ALK fusion gene, or the activation of other oncogenic drivers, which may cause resistance independently of ALK genetic alterations. Pre-clinical data and early clinical trials showed the promising efficacy of a new class of ALK-inhibitors in overcoming acquired resistance. The inhibition of the molecular chaperone, HSP90, represents another promising strategy to overcome crizotinib resistance in ALK-rearranged NSCLC. Several molecules are currently under investigation in order to establish their specific role in the treatment of ALK-rearranged NSCLC.


Expression of mitochondrial regulatory genes parallels respiratory capacity and contractile function in a rat model of hypoxia-induced right ventricular hypertrophy.
Chronic hypobaric hypoxia (CHH) increases load on the right ventricle (RV) resulting in RV hypertrophy. We hypothesized that CHH elicits distinct responses, i.e., the hypertrophied RV, unlike the left ventricle (LV), displaying enhanced mitochondrial respiratory and contractile function. Wistar rats were exposed to 4 weeks CHH (11% O(2)) versus normoxic controls. RV/body weight ratio increased (P < 0.001 vs. control) while RV systolic and developed pressures were higher. However, LV systolic and developed pressures were significantly reduced. Mitochondrial O(2) consumption was sustained in the hypertrophied RV, ADP/O increased (P < 0.01 vs. control) and proton leak significantly decreased. Conversely, LV mitochondrial O(2) consumption was attenuated (P < 0.05 vs. control) and proton leak significantly increased. In parallel, expression of mitochondrial regulators was upregulated in the hypertrophied RV but not the LV. Our data show that the hypertrophied RV induces expression of mitochondrial regulatory genes linking respiratory capacity and enhanced efficiency to sustained contractile function.


Five-nanometer ZnSn2O4:Cr,Eu ultra-small nanoparticles as new near infrared-emitting persistent luminescent nanoprobes for cellular and deep tissue imaging at 800 nm.
Until now, the afterglow emissions of most developed near infrared (NIR)-emitting persistent luminescent nanoparticles (NPLNPs) were located at approximately 700 nm, at the edge of the first tissue transparency window (from 650 to 900 nm), which resulted in relatively low tissue penetration and signal-to-noise ratio (SNR) for in vivo imaging. Herein, 5 nm ZnSn2O4:Cr,Eu (ZSO) NPLNPs with NIR afterglow emission at 800 nm are synthesized via a direct aqueous-phase synthesis method. The longer NIR afterglow emission of ZSO NPLNPs can easily penetrate approximately 3 cm of pork tissue. Furthermore, even though the backbones blocked part of the NIR afterglow light, high SNR (25.5) in vivo images of the backs of mice can be observed and can be maintained for more than 15 min. The ZSO nanoprobes conjugated with folic acid exhibited excellent in vitro and in vivo tumor targeting capacity, which was advantageous for accurate tumor diagnosis. More importantly, the ZSO NPLNPs can be re-excited in situ and in vivo using NIR light to realize renewable near-infrared persistent luminescence in vivo, which was helpful for very long term and higher SNR in vitro and in vivo imaging.


[Long-term outcomes of ICU treatment].
Patients admitted to an intensive care unit (ICU) comprise of a heterogeneous population with substantial differences in admission diagnosis, length of stay and co-morbidity. Therefore, very often the prognosis for each patient differs. In the Netherlands, over 20% of the more than 80,000 patients treated in ICU annually will die within a year of admission. Some of those who survive and are discharged from ICU experience persistent physical, mental and cognitive health problems post-discharge; this is called post-intensive care syndrome (PICS). One year following discharge, circa 50% of patients continue to report physical symptoms, including muscle weakness and walking difficulties. Approximately one in five patients discharged from ICU will develop symptoms akin to post-traumatic stress disorder, and one third will experience depressive symptoms for some time. It remains unclear to what extent the actual ICU admission may potentially contribute to the decline in performance status and quality of life.


The social side of Homo economicus.
Many recent experiments in the field of behavioural economics appear to demonstrate a willingness of humans to behave altruistically, even when it is not in their interest to do so. This has led to the assertion that humans have evolved a special predisposition towards altruism. Recent studies have questioned this, and demonstrated that selfless cooperation does not hold up in controlled experiments. As I discuss here, this calls for more economic 'field experiments' and highlights the need for greater integration of the evolutionary and economic sciences.


The aqueous extract of Withania coagulans fruit partially reverses nicotinamide/streptozotocin-induced diabetes mellitus in rats.
Withania coagulans fruit has been shown to possess antihyperglycemic properties and is used in the traditional Indian system of medicine. However, there has no systematic study of its mechanism of action. In a rat model diabetes mellitus (DM) was induced by intraperitoneal injection of nicotinamide (230 mg/kg of body weight) followed by streptozotocin at 55 mg/kg of body weight. After 96 h, mildly diabetic (MD) (fasting plasma glucose [FPG]=7-11.1 mmol/L) and severely diabetic (SD) (FPG>11.1 mmol/L) rats were treated with aqueous extract of W. coagulans fruit at doses of 125, 250, and 500 mg/kg of body weight/day orally. FPG, postprandial plasma glucose (PPPG), glycosylated hemoglobin (HbA(1c)), plasma insulin, tissue glycogen, and glucose-metabolizing enzymes were assayed at Day 30. Treatment of diabetic animals (MD and SD) with different doses of aqueous W. coagulans resulted in significantly decreased FPG, PPPG, and HbA(1c) (P<.01), whereas serum insulin increased significantly compared with that in diabetic-untreated rats (P<.01). MD and SD animals treated with aqueous W. coagulans also showed significant increases in liver and muscle glycogen compared with diabetic-untreated animals (P<.01). Moreover, activities of glucokinase and phosphofructokinase were also significantly increased (P<.01), whereas glucose-6-phosphatase activity was significantly decreased (P<.01) in MD and SD groups treated with aqueous W. coagulans compared with diabetic-untreated groups. The most effective dose of aqueous W. coagulans was 250 mg/kg of body weight. These results show that the aqueous extract of W. coagulans fruit has significant antihyperglycemic effects, which may be through the modulation of insulin levels and related enzyme activities.


Biodegradable Amphipathic Peptide Hydrogels as Extended-Release System for Opioid Peptides.
Chronic pain is currently treated with opioids that offer unsatisfactory long-term analgesia and produce serious side effects. There is a clear need for alternative therapies. Herein, peptide-based hydrogels are used as extended-release drug delivery carriers. Two different formulations were developed: the drug is coformulated within the hydrogel; the drug is an integral part of the hydrogelator. Both strategies afford a prolonged and significant antinociception up to 72 h after subcutaneous administration in mice.


Glioma-derived mutations in IDH1 dominantly inhibit IDH1 catalytic activity and induce HIF-1alpha.
Heterozygous mutations in the gene encoding isocitrate dehydrogenase-1 (IDH1) occur in certain human brain tumors, but their mechanistic role in tumor development is unknown. We have shown that tumor-derived IDH1 mutations impair the enzyme's affinity for its substrate and dominantly inhibit wild-type IDH1 activity through the formation of catalytically inactive heterodimers. Forced expression of mutant IDH1 in cultured cells reduces formation of the enzyme product, alpha-ketoglutarate (alpha-KG), and increases the levels of hypoxia-inducible factor subunit HIF-1alpha, a transcription factor that facilitates tumor growth when oxygen is low and whose stability is regulated by alpha-KG. The rise in HIF-1alpha levels was reversible by an alpha-KG derivative. HIF-1alpha levels were higher in human gliomas harboring an IDH1 mutation than in tumors without a mutation. Thus, IDH1 appears to function as a tumor suppressor that, when mutationally inactivated, contributes to tumorigenesis in part through induction of the HIF-1 pathway.


The transporters associated with antigen presentation.
Two new members of the ABC superfamily of transporter genes have recently been identified within the Major Histocompatibility Complex in man, rat and mouse. Although the exact function of these genes is not known, they have been shown to be necessary for the presentation of peptides derived from the degradation of cytoplasmic protein antigens to the cellular immune system. For this reason they have been named TAP1 and TAP2 (for Transporter associated with Antigen Presentation). Each gene encodes one membrane spanning domain and one region homologous to the ATP binding domains that characterise the superfamily. The two proteins encoded by the TAP genes form a complex that is localised to the membranes of the endoplasmic reticulum and cis-golgi. Their most likely function is to transport short peptides, that lack signal sequences, from the cytoplasm to the endoplasmic reticulum, although the evidence for this is still indirect.


Size and shape changes during late fetal growth (137-157 gestational days) in the pigtailed macaque (Macaca nemestrina) craniofacial complex: an application using three-dimensional coordinate data and finite element scaling analysis.
Form changes within the fetal pigtailed macaque (Macaca nemestrina) craniofacial complex was documented using finite element scaling analysis (FESA) and three-dimensional (3D) coordinate data for 35 craniofacial landmarks. Coordinate data were digitized from 3D reconstructions of computed tomography (CT) images and 2D axial slices. Twenty-two fetal pigtailed macaques ranging in age from 137 to 157 gestational days were included (in this species, birth is estimated at 170 gestational days). The null hypothesis that the craniofacial complex grows with isometry during late fetal growth of the craniofacial complex was tested (P < 0.05), and the prediction that morphological change along an anteroposterior axis dominates late fetal growth was also investigated. The null hypothesis was rejected, indicating that allometric growth is present during late fetal growth. Growth along an anteroposterior axis is localized in the palate and mandible. The neurocranium grows along a superoinferior axis, while the neurofacial junction displays growth along both the anteroposterior and superoinferior axes. Mediolateral changes are localized between asterions, the external auditory meati, and maxillary and mandibular alveolar points. Finally, a 3D model of craniofacial growth for this species was created, localizing size and shape changes that occur during late fetal growth for each of the 35 craniofacial landmarks defined in this study.


Dynamic in-flight shifts of working memory resources across saccades.
Little is known about how memory resources are allocated in natural vision across sequential eye movements and fixations, as people actively extract information from the visual environment. Here, we used gaze-contingent eye tracking to examine how such resources are dynamically reallocated from old to new information entering working memory. As participants looked sequentially at items, we interrupted the process at different times by extinguishing the display as a saccade was initiated. After a brief interval, participants were probed on one of the items that had been presented. Paradoxically, across all experiments, the final (unfixated) saccade target was recalled more precisely when more items had previously been fixated, that is, with longer rather than shorter saccade sequences. This result is difficult to explain on current models of working memory because recall error, even for the final item, is typically higher as memory load increases. The findings could however be accounted for by a model that describes how resources are dynamically reallocated on a moment-by-moment basis. During each saccade, the target is encoded by consuming a proportion of currently available resources from a limited working memory, as well as by reallocating resources away from previously encoded items. These findings reveal how working memory resources are shifted across memoranda in active vision. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Within-subject variability of Mycobacterium tuberculosis-specific gamma interferon responses in German health care workers.
Gamma interferon (IFN-γ) release assays (IGRAs) are used increasingly for the periodic tuberculosis (TB) screening of health care workers (HCWs), although data regarding the reproducibility and interpretation of serial testing results in countries with a low incidence of TB are scarce. The present study evaluated and compared the within-subject variability of dichotomous and continuous results of two commercial IGRAs, the QuantiFERON-TB Gold In-Tube (QFT) and the T-SPOT.TB (T-SPOT), in German HCWs during a 4-week period. Thirty-five immunocompetent HCWs with low or medium TB screening risk and without known recent TB exposure or tuberculin skin test application were tested repeatedly with both IGRAs at weekly intervals. A total of 158 valid results were obtained for each IGRA. Changes of about ±70% (QFT) and ±60% (T-SPOT) from the mean IFN-γ response accounted for 95% of the within-subject variability. However, according to the manufacturers' cutoffs, inconsistent results were observed more frequently for the QFT (28.6%; four conversions, six reversions) than for the T-SPOT (8.6%; three reversions; P < 0.001). The overall agreement between the IGRAs was good. Regression toward the means accounted for a significant decline in mean IFN-γ responses of about 25% between successive visits for both IGRAs. Although both assays were highly reliable and reproducible, we observed substantial within-subject variability and regression toward the means during a 4-week period, which should be considered when interpreting serial testing results in comparable populations and settings. Our data support the use of borderline zones for the interpretation of serial IGRA results and the retesting of borderline positive results before offering preventive chemotherapy.


Planned Cesarean or planned vaginal delivery for twins: secondary analysis of randomized controlled trial.
To evaluate whether there is a differential benefit of planned Cesarean delivery (CD) over planned vaginal delivery (VD) in women with a twin pregnancy and the first twin in cephalic presentation, depending on prespecified baseline maternal and pregnancy characteristics, and/or gestational age (GA) at delivery. This was a secondary analysis of the Twin Birth Study, which included 2804 women with a twin pregnancy and the first twin (Twin A) in cephalic presentation between 32 + 0 and 38 + 6 weeks' gestation at 106 centers in 25 countries. Women were assigned randomly to either planned CD or planned VD. The main outcome measure was composite adverse perinatal outcome, defined as the occurrence of perinatal mortality or serious neonatal morbidity in at least one twin. The baseline maternal and pregnancy characteristics (markers) considered were maternal age, parity, history of CD, use of antenatal corticosteroids, estimated fetal weight (EFW) of Twin A, EFW of Twin B, > 25% difference in EFW between the twins, presentation of Twin B, chorionicity on ultrasound, method of conception, complications of pregnancy, ruptured membranes at randomization and GA at randomization. Separate logistic regression models were developed for each marker in order to model composite adverse perinatal outcome as a function of the specific marker, planned delivery mode and the interaction between these two terms. In addition, multivariable logistic regression analysis with backward variable elimination was performed separately in each arm of the trial. The association between planned mode of delivery and composite adverse perinatal outcome, according to GA at delivery, was assessed using logistic regression analysis. Of the 2804 women initially randomized, 1391 were included in each study arm. None of the studied baseline markers was associated with a differential benefit of planned CD over planned VD in the rate of composite adverse perinatal outcome. GA at delivery was associated differentially with composite adverse perinatal outcome in the treatment arms (P for interaction < 0.001). Among pregnancies delivered at 32 + 0 to 36 + 6 weeks, there was a trend towards a lower rate of composite adverse perinatal outcome in those in the planned-VD group compared with those in planned-CD group (29 (2.2%) vs 48 (3.6%) cases; odds ratio (OR) 0.62 (95% CI, 0.37-1.03)). In pregnancies delivered at or after 37 + 0 weeks, planned VD was associated with a significantly higher rate of composite adverse perinatal outcome, as compared with planned CD (23 (1.5%) vs 10 (0.7%) cases; OR, 2.25 (95% CI, 1.06-4.77)). The perinatal outcome of twin pregnancies with the first twin in cephalic presentation may differ depending on GA at delivery and planned mode of delivery. At 32-37 weeks, planned VD seems to be favorable, while, from around 37 weeks onwards, planned CD might be safer. The absolute risks of adverse perinatal outcomes at term are low and must be weighed against the increased maternal risks associated with planned CD. © 2019 The Authors. Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.


Potential of the histoculture drug-response assay to contribute to cancer patient survival.
The histoculture drug-response assay (HDRA) was recently evaluated in a retrospective clinical trial and was found to correlate to drug sensitivity, resistance, and patient survival. To further investigate the potential of HDRA to contribute to patient survival, 215 patients with gastric cancer from 45 medical centers were tested with the HDRA in a blinded study after resection of the primary lesion. One hundred sixty-eight patients received at least 20 mg/m2 of mitomycin C and a minimum of 30 g UFT, a mixture of tegafur and uracil at a molar ratio of 1:4, thereby making them eligible for the study. Of these cases 128 were evaluable by the HDRA. The evaluable patient tumors were tested by the HDRA with the [3H]thymidine incorporation end point measured by microautoradiography to be drug "sensitive" or "resistant." The in vitro conditions for distinguishing sensitivity and resistance that matched the response rates for historical controls for gastric carcinoma were 90% inhibition rate and 0.12 microgram/ml for mitomycin C and 70% inhibition rate and 1 microgram/ml for 5-fluorouracil, respectively. Most importantly in the blinded study, the overall and disease-free survival rates of the HDRA-sensitive group were found to be significantly higher than those of the HDRA-resistant group tested under the above conditions. The data further indicate the importance of three-dimensional tumor culture for obtaining accurate clinical information. The results demonstrate that the HDRA response correlates to patient survival, which suggests the potential of the HDRA to contribute to patient survival in gastric cancer when used prospectively.


Induction of Telomere Dysfunction Prolongs Disease Control of Therapy-Resistant Melanoma.
Purpose: Telomerase promoter mutations are highly prevalent in human tumors including melanoma. A subset of patients with metastatic melanoma often fail multiple therapies, and there is an unmet and urgent need to prolong disease control for those patients.Experimental Design: Numerous preclinical therapy-resistant models of human and mouse melanoma were used to test the efficacy of a telomerase-directed nucleoside, 6-thio-2'-deoxyguanosine (6-thio-dG). Integrated transcriptomics and proteomics approaches were used to identify genes and proteins that were significantly downregulated by 6-thio-dG.Results: We demonstrated the superior efficacy of 6-thio-dG both in vitro and in vivo that results in telomere dysfunction, leading to apoptosis and cell death in various preclinical models of therapy-resistant melanoma cells. 6-thio-dG concomitantly induces telomere dysfunction and inhibits the expression level of AXL.Conclusions: In summary, this study shows that indirectly targeting aberrant telomerase in melanoma cells with 6-thio-dG is a viable therapeutic approach in prolonging disease control and overcoming therapy resistance. Clin Cancer Res; 24(19); 4771-84. ©2018 AACR See related commentary by Teh and Aplin, p. 4629.


An updated review of the diagnostic utility of cervical facet joint injections.
Chronic persistent neck pain with or without upper extremity pain is common in the general adult population with a prevalence of 48% for women and 38% for men, with persistent complaints in 22% of women and 16% of men. Multiple modalities of treatment are exploding in managing chronic neck pain along with increasing prevalence. However, there is a paucity of evidence for all modalities of treatments in managing chronic neck pain. Controlled studies have supported the existence of cervical facet or zygapophysial joint pain in 36% to 60% in heterogenous population of these patients. However, these studies also have shown false-positive results in 27% to 63% of patients with a single diagnostic block. A systematic review of diagnostic cervical facet joint nerve blocks. To evaluate and update the accuracy of diagnostic facet joint nerve blocks in the diagnosis of facet joint pain. A methodological quality assessment of included studies was performed using Quality Appraisal of Reliability Studies (QAREL). Only diagnostic accuracy studies meeting at least 50% of the designated inclusion criteria were utilized for analysis. Studies scoring less than 50% are presented descriptively and critically analyzed. The level of evidence was classified as good, fair, and limited or poor based on the quality of evidence developed by the United States Preventive Services Task Force (USPSTF).Data sources included relevant literature identified through searches of PubMed and EMBASE from 1966 to June 2012, and manual searches of the bibliographies of known primary and review articles. Overall, a total of 26 manuscripts were considered for diagnostic accuracy evaluation and 9 manuscripts for studies evaluating various factors influencing the diagnostic validity of facet joint interventions. Based on 9 studies meeting the inclusion criteria utilizing 75% to 100% pain relief as the criterion standard with controlled blocks, the evidence is good for diagnostic accuracy of cervical facet joint pain, with a prevalence of 36% to 60% with a false-positive rate of 27% to 63% with a single block. Based on 2 studies from the same group of authors, the evidence for 75% to 100% pain relief as the criterion standard with a single block is limited. The evidence is limited for a single diagnostic block with 50% to 74% pain relief as the criterion standard, whereas no studies were available assessing the accuracy of 50% to 74% pain relief as the criterion standard with controlled blocks. The limitations of this systematic review include a paucity of literature on outcomes, randomized, placebo-controlled trials and a lack of consensus on a gold standard. Diagnostic cervical facet joint nerve blocks are safe, valid, and reliable. The strength of evidence for diagnostic facet joint nerve blocks is good with the utilization of controlled diagnostic blocks with at least 75% pain relief as the criterion standard; however, the evidence is limited for single blocks or dual blocks for relief of 50% to 74% and single blocks with at least 75% pain relief.


Nocturnal leg cramps: a common complaint in patients with lumbar spinal canal stenosis.
Questionnaire survey on leg cramps for patients with lumbar spinal canal stenosis (LCS). To evaluate the prevalence of leg cramps in patients with LCS treated surgically and the relationship between leg cramps and the surgical outcomes. Although it has been anecdotally reported that LCS patients have suffered from leg cramps, the true prevalence remains unknown. One hundred twenty LCS patients who underwent decompression surgery (men 85, women 35, mean age 73.5) and 370 elderly subjects from the general population (men 162, women 208, mean age 75.6) were enrolled in the study. The participants filled in a questionnaire regarding: (all participants) (1) experience of leg cramps, (2) frequency and time of the day of the cramp attacks; (for LCS patients only), (3) changes in cramps before and after surgery, (4) activities of daily living disturbance because of leg cramps, (5) satisfaction with surgery and walking ability, (6) the Roland-Morris Disability Questionnaire, and (7) the Oswestry Disability Index. Eighty-five (70.8%) patients with LCS and 137 (37.2%) of the control population experienced leg cramps (age and sex adjusted odds ratio; 4.6, P < 0.01). Leg cramps occurred once or twice a week in 34.9% of the LCS group and once in several months in 44.5% of the control group, and occurred nocturnally in 73.3% of the LCS patients and in 91.6% of the control group. In LCS patients, leg cramps improved after surgery in 18.2%, remained unchanged in 45.5%, and worsened in 26.1%, and activities of daily living were disturbed in 47.6%. There was no significant difference in satisfaction with surgery, the Oswestry Disability Index, the Roland-Morris Disability Questionnaire scores, or walking ability between the LCS patients with or without leg cramps. LCS patients had significantly more frequent attacks of nocturnal leg cramps than the control population, and leg cramps disturbed the quality of the patients' life, and they rarely improved after decompression surgery. Leg cramps should be recognized as one of the symptoms of LCS, which disturb the patients' quality of life.


Quantification of microaerobic growth of Geobacter sulfurreducens.
Geobacter sulfurreducens was originally considered a strict anaerobe. However, this bacterium was later shown to not only tolerate exposure to oxygen but also to use it as terminal electron acceptor. Research performed has so far only revealed the general ability of G. sulfurreducens to reduce oxygen, but the oxygen uptake rate has not been quantified yet, nor has evidence been provided as to how the bacterium achieves oxygen reduction. Therefore, microaerobic growth of G. sulfurreducens was investigated here with better defined operating conditions as previously performed and a transcriptome analysis was performed to elucidate possible metabolic mechanisms important for oxygen reduction in G. sulfurreducens. The investigations revealed that cell growth with oxygen is possible to the same extent as with fumarate if the maximum specific oxygen uptake rate (sOUR) of 95 mgO2 gCDW-1 h-1 is not surpassed. Hereby, the entire amount of introduced oxygen is reduced. When oxygen concentrations are too high, cell growth is completely inhibited and there is no partial oxygen consumption. Transcriptome analysis suggests a menaquinol oxidase to be the enzyme responsible for oxygen reduction. Transcriptome analysis has further revealed three different survival strategies, depending on the oxygen concentration present. When prompted with small amounts of oxygen, G. sulfurreducens will try to escape the microaerobic area; if oxygen concentrations are higher, cells will focus on rapid and complete oxygen reduction coupled to cell growth; and ultimately cells will form protective layers if a complete reduction becomes impossible. The results presented here have important implications for understanding how G. sulfurreducens survives exposure to oxygen.


Evaluation of Diffuse Reflection Infrared Spectrometry for End-of-Shift Measurement of α-quartz in Coal Dust Samples.
The inhalation of toxic substances is a major threat to the health of miners, and dust containing respirable crystalline silica (α-quartz) is of particular concern, due to the recent rise in cases of coal workers' pneumoconiosis and silicosis in some U.S. mining regions. Currently, there is no field-portable instrument that can measure airborne α-quartz and give miners timely feedback on their exposure. The U.S. National Institute for Occupational Safety and Health (NIOSH) is therefore conducting studies to investigate technologies capable of end-of-shift or real-time measurement of airborne quartz. The present study focuses on the potential application of Fourier transform infrared (FT-IR) spectrometry conducted in the diffuse reflection (DR) mode as a technique for measuring α-quartz in respirable mine dust. A DR accessory was used to analyze lab-generated respirable samples of Min-U-Sil 5 (which contains more than 90% α-quartz) and coal dust, at mass loadings in the ranges of 100-600 μg and 600-5300 μg, respectively. The dust samples were deposited onto three different types of filters, borosilicate fiberglass, nylon, and polyvinyl chloride (PVC). The reflectance, R, was calculated by the ratio of a blank filter and a filter with deposited mine dust. Results suggest that for coal and pure quartz dusts deposited on 37 mm PVC filters, measurements of -log R correlate linearly with known amounts of quartz on filters, with R(2) values of approximately 0.99 and 0.94, respectively, for samples loaded up to ∼4000 μg. Additional tests were conducted to measure quartz in coal dusts deposited onto the borosilicate fiberglass and nylon filter media used in the NIOSH-developed Personal Dust Monitor (PDM). The nylon filter was shown to be amenable to DR analysis, but quantification of quartz is more accurate when the filter is "free," as opposed to being mounted in the PDM filter holder. The borosilicate fiberglass filters were shown to produce excessive interference, making quartz quantification impossible. It was concluded that, while the DR/FT-IR method is potentially useful for on-filter measurement of quartz in dust samples, the use of PVC filters produced the most accurate results.


Developing a new interdisciplinary lab course for undergraduate and graduate students: Plant cells and proteins.
Studies of protein function increasingly use multifaceted approaches that span disciplines including recombinant DNA technology, cell biology, and analytical biochemistry. These studies rely on sophisticated equipment and methodologies including confocal fluorescence microscopy, mass spectrometry, and X-ray crystallography that are beyond the scope of traditional laboratory courses. To equip the advanced undergraduate and beginning graduate students with an enabling base of knowledge and initial experience with advanced protein research methodologies, a laboratory course entitled Plant Cells and Proteins was developed in a partnership between Washington University and the Donald Danforth Plant Science Center in St. Louis. In this one semester course, 10-12 students obtain hands-on experience with plant tissue culture, gene transformation, subcellular localization of fluorescent recombinant proteins using confocal microscopy, purification of affinity-tagged recombinant proteins, isolation of total protein extracts, enzymatic assays, one- and two-dimensional gel electrophoresis, MALDI-TOF and ESI-Q-TOF mass spectrometry, protein crystallization, and X-ray diffraction. The course is taught as a series of modules, each led by an expert researcher. Students are evaluated based on a series of graded written reports and tests of their mastery of key concepts, interpretations, and the limitations of the experimental methods.


Mutational and functional analysis reveals ADAMTS18 metalloproteinase as a novel driver in melanoma.
The disintegrin-metalloproteinases with thrombospondin domains (ADAMTS) genes have been suggested to function as tumor suppressors as several have been found to be epigenetically silenced in various cancers. We performed a mutational analysis of the ADAMTS gene family in human melanoma and identified a large fraction of melanomas to harbor somatic mutations. To evaluate the functional consequences of the most commonly mutated gene, ADAMTS18, six of its mutations were biologically examined. ADAMTS18 mutations had little effect on melanoma cell growth under standard conditions, but reduced cell dependence on growth factors. ADAMTS18 mutations also reduced adhesion to laminin and increased migration in vitro and metastasis in vivo. Melanoma cells expressing mutant ADAMTS18 had reduced cell migration after short hairpin RNA-mediated knockdown of ADAMTS18, suggesting that ADAMTS18 mutations promote growth, migration, and metastasis in melanoma.


A case-control study to investigate the risk of leukaemia associated with exposure to benzene in petroleum marketing and distribution workers in the United Kingdom.
To investigate the risk of leukaemia in workers in the petroleum distribution industry who were exposed to low levels of benzene. From the cohort of distribution workers, 91 cases were identified as having leukaemia on either a death certificate or on cancer registration. These cases were compared with controls (four per case) randomly selected from the cohort, who were from the same company as the respective case, matched for age, and alive and under follow up at the time of case occurrence. Work histories were collected for the cases and controls, together with information about the terminals at which they had worked, fuel compositions, and occupational hygiene measurements of benzene. These data were used to derive quantitative estimates of personal exposure to benzene. Odds ratios (OR) were calculated conditional on the matching, to identify those variables in the study which were associated with risk of leukaemia. Examination of the potential effects of confounding and other variables was carried out with conditional logistic regression. Analyses were carried out for all leukaemia and separately for acute lymphoblastic, chronic lymphocytic, acute myeloid and monocytic, and chronic myeloid leukaemias. There was no significant increase in the overall risk of all leukaemias with higher cumulative exposure to benzene or with intensity of exposure, but risk was consistently doubled in subjects employed in the industry for > 10 years. Acute lymphoblastic leukaemia tended to occur in workers employed after 1950, who started work after the age of 30, worked for a short duration, and experienced low cumulative exposure with few peaks. The ORs did not increase with increasing cumulative exposure. The risk of chronic lymphocytic leukaemia seemed to be related most closely to duration of employment and the highest risk occurred in white collar workers with long service. These workers had only background levels of benzene exposure. There was no evidence of an association of risk with any exposure variables, and no evidence of an increasing risk with increasing cumulative exposure, mean intensity, or maximum intensity of exposure. The patterns of risk for acute myeloid and monocytic leukaemia were different from those of the lymphoid subgroups, in which duration of employment was the variable most closely related to risk. Risk was increased to an OR of 2.8 (95% confidence interval (95% CI) 0.8 to 9.4) for a cumulative exposure between 4.5 and 45 ppm-years compared with < 0.45 ppm-years. For mean intensity between 0.2 and 0.4 ppm an OR of 2.8 (95% CI 0.9 to 8.5) was found compared with < 0.02 ppm. Risk did not increase with cumulative exposure, maximum intensity, or mean intensity of exposure when treated as continuous variables. Cases of acute myeloid and monocytic leukaemia were more often classified as having peaked exposures than controls, and when variables characterising peaks, particularly daily and weekly peaks, were included in the analysis these tended to dominate the other exposure variables. However, because of the small numbers it is not possible to distinguish the relative influence of peaked and unpeaked exposures on risk of acute myeloid and monocytic leukaemia. There was no evidence of an increased risk of chronic myeloid leukaemia with increases in cumulative exposure, maximum intensity, mean intensity, and duration of employment, either as continuous or categorical variables. Analyses exploring the sensitivity of the results to the source and quality of the work histories showed similar patterns in general. However, no increases in ORs for categories of cumulative exposure were found for acute myeloid and monocytic leukaemia in the data set which included work histories obtained from personnel records still in existence, although numbers were reduced. Analyses excluding the last five and 10 years of exposure showed a tendency for ORs to reduce for chronic lymphocytic leukaemia and chronic myeloid leukaemia, and to increase for acute myeloid and monocytic leukaemia. Limitations of the study include uncertainties and gaps in the information collected, and small numbers in subcategories of exposure which can lead to wide CIs around the risk estimates and poor fit of the mathematical models. There is no evidence in this study of an association between exposure to benzene and lymphoid leukaemia, either acute or chronic. There is some suggestion of a relation between exposure to benzene and myeloid leukaemia, in particular for acute myeloid and monocytic leukaemia. Peaked exposures seemed to be experienced for this disease. However, in view of the limitations of the study, doubt remains as to whether the risk of acute myeloid and monocytic leukaemia is increased by cumulative exposures of < 45 ppm-years. Further work is recommended to review the work histories and redefine their quality, to explore the discrepancies between results for categorical and continuous variables, and to develop ranges around the expose estimates to enable further sensitivity analyses to be carried out.


Hemolysis during cardiac extracorporeal membrane oxygenation: a case-control comparison of roller pumps and centrifugal pumps in a pediatric population.
Extracorporeal membrane oxygenation (ECMO) is a lifesaving therapy, which has been used for the support of children with a broad range of diseases. Two pumps of differing mechanisms have been used to generate the extracorporeal flow: roller-head pumps and centrifugal pumps. Seven patients supported during ECMO with Levitronix Centrimag (Centrimag group [CG]) were matched to 14 patients supported with Stockert-Shiley SIII (Stockert-Shiley group [SSG]) at a single institution from July 2007 to July 2009. We hypothesized that hemolysis as measured by plasma-free hemoglobin (PFH) is elevated in the SSG versus the CG during cardiac ECMO. Categorical data were analyzed using Fisher's exact test. Plasma-free hemoglobin differences between groups were analyzed using both Wilcoxon rank sum and beta regression. Overall, SSG patients had two times the odds of having a higher PFH than CG patients adjusting for repeated measures (odds ratio [OR] = 1.96, 95% confidence interval [CI]: [1.15-3.34], p < 0.014). Differences between circuit failure in the first 168 hours did not reach statistical significance (1/7 CG vs. 7/14 SSG; p = 0.174). In this population of cardiac patients requiring ECMO support, more hemolysis occurred in the SSG, a roller-head pump supported group, when compared with the CG, a centrifugal pump supported group. Differences in circuit life did not reach statistical significance. This pilot study contrasts with past studies, which have demonstrated more hemolysis occurring with centrifugal pumps when compared with roller-head pumps.