A comparison of macrophage colony-stimulating factor (M-CSF) gene expression in primary and immortalized endothelial cells.
M-CSF is produced by a wide variety of cell types, including EC, fibroblasts, and monocyte/macrophages, where it functions as a survival factor and a chemotactic agent for monocytes. An early event in the development of atherosclerosis is the infiltration of monocytes into the artery wall. Local expression of M-CSF by EC lining the blood vessels is thought to promote the growth and survival of lesional monocytes and macrophages, thus enhancing lesion development and disease progression. Primary cultures of EC are difficult to maintain for long periods of time, which complicates their use for biochemical and molecular analysis. As a step toward identifying a representative endothelial-like cell line, serum-dependent and IL-1-dependent changes in M-CSF gene expression in two endothelial-like cell lines were compared to that detected in primary EC cultures. The data presented here demonstrate that the two endothelial-like cell lines, like primary cultures of EC, express the M-CSF gene under basal conditions. In both types of cell cultures, IL-1alpha stimulation increased M-CSF mRNA levels 2-7-fold, whereas serum stimulation elicited a more modest effect (2-3-fold increase). The IL-1alpha-induced change in M-CSF gene expression is mediated at the transcriptional level, and M-CSF promoter activity is, in part, dependent on the activity of the NF-kappaB-inducing kinase. Collectively, our results demonstrate that either endothelial-like cell line would be a representative model in which endothelial-specific changes in M-CSF gene expression could be identified.


Combined Analysis of RNA Sequence and Microarray Data Reveals a Competing Endogenous RNA Network as Novel Prognostic Markers in Malignant Pleural Mesothelioma.
Malignant pleural mesothelioma (MPM) is a highly aggressive cancer with short survival time. Unbalanced competing endogenous RNAs (ceRNAs) have been shown to participate in the tumor pathogenesis and served as biomarkers for the clinical prognosis. However, the comprehensive analyses of the ceRNA network in the prognosis of MPM are still rarely reported. In this study, we obtained the transcriptome data of the MPM and the normal samples from TCGA, EGA, and GEO databases and identified the differentially expressed (DE) mRNAs, lncRNAs, and miRNAs. The functions of the prognostic genes and the overlapped DEmRNAs were further annotated by the multiple enrichment analyses. Then, the targeting relationships among lncRNA-miRNA and miRNA-mRNA were predicted and calculated, and a prognostic ceRNA regulatory network was established. We included the prognostic 73 mRNAs and 13 miRNAs and 26 lncRNAs into the ceRNA network. Moreover, 33 mRNAs, three miRNAs, and seven lncRNAs were finally associated with prognosis, and a model including seven mRNAs, two lincRNAs, and some clinical factors was finally established and validated by two independent cohorts, where CDK6 and SGMS1-AS1 were significant to be independent prognostic factors. In addition, the identified co-expressed modules associated with the prognosis were overrepresented in the ceRNA network. Multiple enrichment analyses showed the important roles of the extracellular matrix components and cell division dysfunction in the invasion of MPM potentially. In summary, the prognostic ceRNA network of MPM was established and analyzed for the first time and these findings shed light on the function of ceRNAs and revealed the potential prognostic and therapeutic biomarkers of MPM.


Potential effect of herbs on diabetic hypertension: alternative medicine treatment modalities.
Diabetes mellitus is commonly known to lead to life threatening complications. Among them, hypertension is one complication which may be highlighted. In modern medical society, several drugs and treatment regimen have developed to treat diabetes mellitus. Although they obtain positive impacts, yet there are certain limitations encountered in the management of the disease due to their adverse effects and non-compliance by the patients. Herbs have been valued as a supplement in treating chronic oxidative stress disorder like diabetes mellitus. In the present review, some of the herbs which possess both anti-hyperglycemic and anti-hypertensive properties are being highlighted. Potential herbs which include Andrographis Paniculata, Ammi Visnaga, Allium sativum, Phyllanthus amarus, Ginkgo biloba, Solanum tuberosum, Tuberosum L. Piper sarmentosum and Lamiaceae family are known to possess antihypertensive properties. Various researches have been conducted on these herbs and positive results have been obtained. Based on these results, the present review article discusses the alternative management of diabetic hypertension with the herbal medicine. In conclusion, the present review article stresses on the need to ensure global awareness about the traditional medicines while treating diabetic hypertension.


[Effect of microbial aerosols on the human].
Based on numerous publications dealing with the effects of microbial emissions on workers in waste processing plants and on few papers concerning the assessment of the environmental health relevance of microbial aerosols on people living in the vicinity of composting facilities, the current state of the art in this field is presented. With regard to occupational exposure the possible health effects like infections, toxicity and allergies are specified. Since to date only few studies have been made of populations exposed to microbial aerosols in ambient air, the environmental health aspects are reviewed in the context of a study of three compost plants in Hesse, where ambient air measurements as well as epidemiological investigations were carried out. Final recommendations are given and the research needs regarding the environmental health significance of microbial aerosols are formulated.


Appropriate first-line regimens to combat Helicobacter pylori antibiotic resistance: an Asian perspective.
Asia has the largest population of any continent and the highest incidence of gastric cancer in the world, making it very important in the context of Helicobacter pylori infection. According to current guidelines, standard triple therapy containing a proton pump inhibitor (PPI) and two antibiotics; amoxicillin (AMX) and clarithromycin (CAM) or metronidazole (MNZ), is still the preferred first-line regimen for treatment of H. pylori infection. However, the efficacy of legacy triple regimens has been seriously challenged, and they are gradually becoming ineffective. Moreover, some regions in Asia show patterns of emerging antimicrobial resistance. More effective regimens including the bismuth and non-bismuth quadruple, sequential, and dual-concomitant (hybrid) regimens are now replacing standard triple therapies as empirical first-line treatments on the basis of the understanding of the local prevalence of H. pylori antimicrobial resistance. Selection of PPI metabolized by the non-enzymatic pathway or minimal first pass metabolism and/or increasing dose of PPI are important to increase H. pylori eradication rates. Therefore, local antibiotic resistance surveillance updates, selection of appropriate first-line regimens with non-enzymatic PPI and/or increased doses of PPI, and detailed evaluation of patients' prior antibiotic usage are all essential information to combat H. pylori antibiotic resistance in Asia.


Genome-wide identification and functional analysis of genes expressed ubiquitously in rice.
Genes that are expressed ubiquitously throughout all developmental stages are thought to be necessary for basic biological or cellular functions. Therefore, determining their biological roles is a great challenge. We identified 4034 of these genes in rice after studying the results of Agilent 44K and Affymetrix meta-anatomical expression profiles. Among 105 genes that were characterized by loss-of-function analysis, 79 were classified as members of gene families, the majority of which were predominantly expressed. Using T-DNA insertional mutants, we examined 43 genes and found that loss of expression of six genes caused developing seed- or seedling-defective phenotypes. Of these, three are singletons without similar family members and defective phenotypes are expected from mutations. Phylogenomic analyses integrating genome-wide transcriptome data revealed the functional dominance of three ubiquitously expressed family genes. Among them, we investigated the function of Os03g19890, which is involved in ATP generation within the mitochondria during endosperm development. We also created and evaluated functional networks associated with this gene to understand the molecular mechanism. Our study provides a useful strategy for pheonome analysis of ubiquitously expressed genes in rice.


Detection of mutations in the DNA polymerase delta gene of human sporadic colorectal cancers and colon cancer cell lines.
To test the hypothesis whether DNA polymerases acquire mutator properties during tumor development (mutator hypothesis), we examined DNA polymerase delta mRNA in 6 colon cancer cell lines (DLD-1, HCT116, SW48, HT29, SW480 and SW620) and 7 sporadic human colorectal cancers. For analysis we used amplification of cDNA by polymerase chain reaction, single-strand conformation polymorphism and sequencing techniques. In 5 of the cell lines, 9 mutations leading to changes of the amino acid sequence of DNA polymerase delta were detected. Most mutations were found in the cell lines DLD-1, HCT116 and SW48 for which defects in mismatch repair genes had been identified previously. In the majority of cases, wild type and mutated sequences were present. In 2 cell lines (HCT116 and SW48), a single-nucleotide deletion occurred at the same position. This resulted in a premature termination codon by which the DNA interaction domain of the enzyme was eliminated. Furthermore, sequence deviations were found in the tumor tissues of 4 colon cancer patients. Wild-type and altered sequences were present simultaneously. The deviations included missense mutations (2 cases) and silent mutations (2 cases). The missense mutations and one of the silent mutations were found in normal mucosa as well. In addition, the mutation clustered region of a tumor suppressor gene, often found to be defective in colon cancer, the adenomatous polyposis coli (APC) gene, was investigated in surgical specimens and cell lines. One carcinoma and 2 cell lines exhibited amino acid changes in both the DNA polymerase delta gene and in the mutation clustered region of the APC gene. Since most of the mutations detected in the DNA polymerase delta mRNA are likely to alter the structure of the protein, the enzyme is expected to be functionally impaired. In particular, copying fidelity might be decreased, thus contributing to the high mutation rate observed in colorectal cancer.


FUNDC1/PFKP-mediated mitophagy induced by KD025 ameliorates cartilage degeneration in osteoarthritis.
Osteoarthritis (OA) is the most common joint disease, but no disease-modifying drugs have been approved for OA treatment. Mitophagy participates in mitochondrial homeostasis regulation by selectively clearing dysfunctional mitochondria, which might contribute to cartilage degeneration in OA. Here, we provide evidence of impaired mitophagy in OA chondrocytes, which exacerbates chondrocyte degeneration. Among the several classic mitophagy-regulating pathways and receptors, we found that FUNDC1 plays a key role in preserving chondrocyte homeostasis by inducing mitophagy. FUNDC1 knockdown in vitro and knockout in vivo decreased mitophagy and exacerbated mitochondrial dysfunction, exacerbating chondrocyte degeneration and OA progression. FUNDC1 overexpression via intra-articular injection of adeno-associated virus alleviated cartilage degeneration in OA. Mechanistically, our study demonstrated that PFKP interacts with and dephosphorylates FUNDC1 to induce mitophagy in chondrocytes. Further analysis identified KD025 as a candidate drug for restoring chondrocyte mitophagy by increasing the FUNDC1-PFKP interaction and thus alleviating cartilage degeneration in mice with DMM-induced OA. Our study highlights the role of the FUNDC1-PFKP interaction in chondrocyte homeostasis via mitophagy induction and identifies KD025 as a promising agent for treating OA by increasing chondrocyte mitophagy.


Calcium dynamics regulating the timing of decision-making in C. elegans.
Brains regulate behavioral responses with distinct timings. Here we investigate the cellular and molecular mechanisms underlying the timing of decision-making during olfactory navigation in Caenorhabditis elegans. We find that, based on subtle changes in odor concentrations, the animals appear to choose the appropriate migratory direction from multiple trials as a form of behavioral decision-making. Through optophysiological, mathematical and genetic analyses of neural activity under virtual odor gradients, we further find that odor concentration information is temporally integrated for a decision by a gradual increase in intracellular calcium concentration ([Ca2+]i), which occurs via L-type voltage-gated calcium channels in a pair of olfactory neurons. In contrast, for a reflex-like behavioral response, [Ca2+]i rapidly increases via multiple types of calcium channels in a pair of nociceptive neurons. Thus, the timing of neuronal responses is determined by cell type-dependent involvement of calcium channels, which may serve as a cellular basis for decision-making.


Investigation of pseudo-urogenital myiasis and errors in diagnosis.
Cases of genitourinary myiasis in humans caused by larvae of flies are reported from time to time. In our study, 11 patients, who reported worms in their urine and brought these worms to our laboratory, were investigated. The larvae were found to belong to the subfamily Psychodinae. By taking urine samples carefully from the patients, examining and evaluating their history, it was concluded that they did not actually have myiasis. We endeavour to describe the pitfalls in this diagnosis.


Are one's attachment avoidance toward a particular person and his/her placement of this particular person in the attachment hierarchy inversely overlapping? Four bifactor-analysis studies.
Do one's hierarchical preference for attachment support from a particular person over other people (attachment hierarchy) and his/her discomfort with closeness and uneasiness about being dependent on that particular person (attachment avoidance) inversely overlap? These two constructs have been distinctly conceptualized. Attachment hierarchy has been regarded as a normative characteristic of attachment relationships, while attachment avoidance has been considered to reflect an individual difference of relationship quality. Employing bifactor analyses, we demonstrated a unidimensional general factor of these two concepts in four studies exploring Czech young adults' relationships with mother, father, friends, and romantic partner (Study 1); U.S. young adults' relationships with a romantic partner (Study 2); Czech adolescents' relationships with mother, father, and friends (Study 3); and Japanese young adults' relationships with mother, father, and romantic partner (Study 4). These convergent results provide the replicable and generalizable evidence that one's attachment avoidance toward a particular person and her/his placement of that particular person in the attachment hierarchy are inversely overlapping.


[Chorea subsequent to acute rheumatic fever in a 9-year-old girl].
A nine-year-old girl had acute choreatic symptoms in her face and limbs, after a throat infection 6 weeks previously. On auscultation of the heart a systolic murmur was found and echocardiography showed mitral valve incompetence. There was a positive anti-deoxyribonuclease B titre in the serum, providing evidence of a previously contracted streptococcal infection. Both chorea and acquired carditis are major criteria for the diagnosis of acute rheumatic fever. The course was characterized--as it usually is--by spontaneous, gradual resolution of the symptoms. Protracted penicillin prophylaxis is indicated to prevent recurrence of acute rheumatic fever and cardiac valvular damage.


Differences between young adults and elderly in thermal comfort, productivity, and thermal physiology in response to a moderate temperature drift and a steady-state condition.
Results from naturally ventilated buildings show that allowing the indoor temperature to drift does not necessarily result in thermal discomfort and may allow for a reduction in energy use. However, for stationary conditions, several studies indicate that the thermal neutral temperature and optimum thermal condition differ between young adults and elderly. There is a lack of studies that describe the effect of aging on thermal comfort and productivity during a moderate temperature drift. In this study, the effect of a moderate temperature drift on physiological responses, thermal comfort, and productivity of eight young adults (age 22-25 year) and eight older subjects (age 67-73 year) was investigated. They were exposed to two different conditions: S1-a control condition; constant temperature of 21.5 degrees C; duration: 8 h; and S2-a transient condition; temperature range: 17-25 degrees C, duration: 8 h, temperature drift: first 4 h: +2 K/h, last 4 h: -2 K/h. The results indicate that thermal sensation of the elderly was, in general, 0.5 scale units lower in comparison with their younger counterparts. Furthermore, the elderly showed more distal vasoconstriction during both conditions. Nevertheless, TS of the elderly was related to air temperature only, while TS of the younger adults also was related to skin temperature. During the constant temperature session, the elderly preferred a higher temperature in comparison with the young adults. Because the stock of fossil fuels is limited, energy savings play an important role. Thermal comfort is one of the most important performance indicators to successfully apply measures to reduce the energy need in buildings. Allowing drifts in indoor temperature is one of the options to reduce the energy demand. This study contributes to the knowledge concerning the effects of a moderate temperature drift and the age of the inhabitants on their thermal comfort.


Experimental entanglement activation from discord in a programmable quantum measurement.
In quantum mechanics, observing is not a passive act. Consider a system of two quantum particles A and B: if a measurement apparatus M is used to make an observation on B, the overall state of the system AB will typically be altered. When this happens, no matter which local measurement is performed, the two objects A and B are revealed to possess peculiar correlations known as quantum discord. Here, we demonstrate experimentally that the very act of local observation gives rise to an activation protocol which converts discord into distillable entanglement, a stronger and more useful form of quantum correlations, between the apparatus M and the composite system AB. We adopt a flexible two-photon setup to realize a three-qubit system (A, B, M) with programmable degrees of initial correlations, measurement interaction, and characterization processes. Our experiment demonstrates the fundamental mechanism underpinning the ubiquitous act of observing the quantum world and establishes the potential of discord in entanglement generation.


Critical evaluation of the effect of valerian extract on sleep structure and sleep quality.
A carefully designed study assessed the short-term (single dose) and long-term (14 days with multiple dosage) effects of a valerian extract on both objective and subjective sleep parameters. The investigation was performed as a randomised, double-blind, placebo-controlled, cross-over study. Sixteen patients (4 male, 12 female) with previously established psychophysiological insomnia (ICSD-code 1.A.1.), and with a median age of 49 (range: 22 to 55), were included in the study. The main inclusion criteria were reported primary insomnia according to ICSD criteria, which was confirmed by polysomnographic recording, and the absence of acute diseases. During the study, the patients underwent 8 polysomnographic recordings: i.e., 2 recordings (baseline and study night) at each time point at which the short and long-term effects of placebo and valerian were tested. The target variable of the study was sleep efficiency. Other parameters describing objective sleep structure were the usual features of sleep-stage analysis, based on the rules of Rechtschaffen and Kales (1968), and the arousal index (scored according to ASDA criteria, 1992) as a sleep microstructure parameter. Subjective parameters such as sleep quality, morning feeling, daytime performance, subjectively perceived duration of sleep latency, and sleep period time were assessed by means of questionnaires. After a single dose of valerian, no effects on sleep structure and subjective sleep assessment were observed. After multiple-dose treatment, sleep efficiency showed a significant increase for both the placebo and the valerian condition in comparison with baseline polysomnography. We confirmed significant differences between valerian and placebo for parameters describing slow-wave sleep. In comparison with the placebo, slow-wave sleep latency was reduced after administration of valerian (21.3 vs. 13.5 min respectively, p<0.05). The SWS percentage of time in bed (TIB) was increased after long-term valerian treatment, in comparison to baseline (9.8 vs. 8.1% respectively, p<0.05). At the same time point, a tendency for shorter subjective sleep latency, as well as a higher correlation coefficient between subjective and objective sleep latencies, were observed under valerian treatment. Other improvements in sleep structure - such as an increase in REM percentage and a decrease in NREM1 percentage - took place simultaneously under placebo and valerian treatment. A remarkable finding of the study was the extremely low number of adverse events during the valerian treatment periods (3 vs. 18 in the placebo period). In conclusion, treatment with a herbal extract of radix valerianae demonstrated positive effects on sleep structure and sleep perception of insomnia patients, and can therefore be recommended for the treatment of patients with mild psychophysiological insomnia.


Templated DNA ligation with thiol chemistry.
We describe two DNA-templated ligation strategies: native chemical ligation (NCL), and thiol-disulfide exchange. Both systems result in successful ligation in the presence of a DNA template. The stability of the product from the NCL reaction relies on exogenous thiol, while the thiol-disulfide reaction proceeds in a catalyst-free manner.


Discovery of Novel Small-Molecule Inhibitors of LIM Domain Kinase for Inhibiting HIV-1.
A dynamic actin cytoskeleton is necessary for viral entry, intracellular migration, and virion release. For HIV-1 infection, during entry, the virus triggers early actin activity by hijacking chemokine coreceptor signaling, which activates a host dependency factor, cofilin, and its kinase, the LIM domain kinase (LIMK). Although knockdown of human LIM domain kinase 1 (LIMK1) with short hairpin RNA (shRNA) inhibits HIV infection, no specific small-molecule inhibitor of LIMK has been available. Here, we describe the design and discovery of novel classes of small-molecule inhibitors of LIMK for inhibiting HIV infection. We identified R10015 as a lead compound that blocks LIMK activity by binding to the ATP-binding pocket. R10015 specifically blocks viral DNA synthesis, nuclear migration, and virion release. In addition, R10015 inhibits multiple viruses, including Zaire ebolavirus (EBOV), Rift Valley fever virus (RVFV), Venezuelan equine encephalitis virus (VEEV), and herpes simplex virus 1 (HSV-1), suggesting that LIMK inhibitors could be developed as a new class of broad-spectrum antiviral drugs.IMPORTANCE The actin cytoskeleton is a structure that gives the cell shape and the ability to migrate. Viruses frequently rely on actin dynamics for entry and intracellular migration. In cells, actin dynamics are regulated by kinases, such as the LIM domain kinase (LIMK), which regulates actin activity through phosphorylation of cofilin, an actin-depolymerizing factor. Recent studies have found that LIMK/cofilin are targeted by viruses such as HIV-1 for propelling viral intracellular migration. Although inhibiting LIMK1 expression blocks HIV-1 infection, no highly specific LIMK inhibitor is available. This study describes the design, medicinal synthesis, and discovery of small-molecule LIMK inhibitors for blocking HIV-1 and several other viruses and emphasizes the feasibility of developing LIMK inhibitors as broad-spectrum antiviral drugs.


Insulin up-regulates vascular endothelial growth factor and stabilizes its messengers in endometrial adenocarcinoma cells.
Angiogenesis is crucial for tumor growth and dissemination. Vascular endothelial growth factor (VEGF) is a potent angiogenic factor that promotes vascular growth and therefore tumoral growth and metastasis. Overweight, frequently associated with hyperinsulinemia, constitutes the major risk factor for endometrial carcinoma. Thus, elevated insulin levels may partly explain the increased risk of endometrial cancer observed in obese postmenopausal women. The aim of the present work was to test the role of insulin in the control of VEGF expression in endometrial carcinoma cells (HEC-1A). We have shown that insulin induced a biphasic expression of VEGF messenger ribonucleic acid, with an early, but low, induction (4 h of stimulation) and a delayed, but high, induction (24 h). The delayed effect of insulin on VEGF expression involved transcriptional and posttranscriptional regulation, as evidenced by the increased rate of VEGF transcription and the prolonged half-life of VEGF messenger ribonucleic acid. Simultaneously we observed higher levels of VEGF protein in the conditioned medium of stimulated cells compared with unstimulated ones. Therefore, insulin could contribute to the increased risk of endometrial carcinoma due to its ability to induce VEGF expression and thus participate in the maintenance of an angiogenic phenotype.


Chelation in metal intoxication. XLIV: Efficacy of alpha-mercapto-beta-(5-substituted, 2-furyl) acrylic acids in mobilizing intracellularly bound cadmium in rat.
The efficacy of alpha-mercapto-beta-(2-furyl) acrylic acid (MFA), alpha-mercapto-beta-(5-sodiumsulfonate, 2-furyl) acrylic acid (MSFA) and alpha-mercapto-beta-(5-acetoxymethyl, 2-furyl) acrylic acid (MAFA) to mobilize intracellularly bound cadmium in liver and kidney was investigated in rats pre-exposed to cadmium. MFA was effective in reducing cadmium levels of hepatic and renal supernatant cytosolic fraction (SCF) while MSFA and MAFA were effective in lowering cadmium levels of renal SCF and hepatic SCF respectively. All the chelating agents also enhanced the excretion of cadmium more in feces than in urine. However, substitution on the furan ring lowered cadmium mobilizing efficacy of the parent compound, MFA. The treatment with MFA did not affect the status of endogenous zinc and copper while the treatment with MSFA and MAFA enhanced their excretion. MSFA increased hepatic and renal zinc and renal copper while MAFA increased their copper levels.


[Actiprotective and antihypoxic action of new heteroaromatic antioxidants].
Expernents with mice showed that most of 15 new heteroaromatic antioxidant compounds possess aciprotective and antixopixic properties. Based on results of treadmill and swimming tests, actiprotective action of IBKhF-1, 11 and 14 surpassed greatly bemythil and bromanthane in ordinary conditions. Inhibitor of gluconeogenase tryptophan cancelled largely the stimulatting action of highly effective and active IBKhF-1, 2 and 11 on physical performance during treadmill exercise. Consequently, gluconeogenesis activation is one of the major components of the actiprotective action of these antioxidants. In addition, IBKhF-1, 11 and 14 excelled bemythil and bromanthane in the extreme conditions of running in hyperthermia and swimming in acute hypoxia combined with hypercapnia. IBKhF-2 and 14 were better than amtisol (standard antihypoxic agent) and bemythil against acute hypoxia in pressure chamber, whereas IBKhF-4 and 14 excelled these agents in thermal chamber.


Estimation and identification in blood plasma of paracetamol (N-acetyl-p-aminophenol) in the presence of barbiturates.
Barbiturates are a source of interference in the estimation of paracetamol by both UV spectrophotometry and gas-liquid chromatography (GLC). Experiments have shown that the interference by barbiturates on GLC can be avoided by benzoylation (in aqueous solution) of the paracetamol. This O-benzoyl derivative can be N-silylated to give an additional identification parameter. The method, which requires ml of sample, will quantitatively and qualitatively estimate "free" paracetamol in plasma and in post mortem blood in the range of 2-40 mug of drug per millilitre of sample.


Preparation and characterization of monoclonal antibody against melatonin.
Anti-melatonin monoclonal antibodies (MAb) were prepared following coupling melatonin to bovine serum albumin (BSA) by Mannich reaction. Balb/c mice were immunized via injection of the melatonin-BSA intraperitonally. The spleen cells producing high titer of antibody were fused with myeloma cells of SP2/0 origin. After two limiting dilutions, two stable clones (AS-H10 and AS-D26) exhibiting best properties were selected for further studies. The class and subclass of two MAbs were found to be IgG(1) and IgG(2a) with lambda and kappa light chains, respectively. Antibodies secreted by these two clones showed high affinity of about 10(9)M(1). Study of the specificity criteria showed that these clones had no cross reactivity with indolic, aromatic, and imidazole ring-containing compounds, and had high specificity towards melatonin. The calibration curve was constructed with a sensitivity range of 10 ng/mL to 10 microg/mL. In conclusion, these MAbs may be useful for immunoassay of melatonin.


MOOCs and Medical Education: Hope or Hype? .
This article was migrated. The article was marked as recommended. The use of Massive Open Online Courses (MOOCs) in medical education is evolving and the potential of these learning and teaching resources has not yet been fully explored. Having previously integrated MOOCs within our medical undergraduate curriculum in a variety of contexts, in this study we focused on the attitudes of staff and students to the perceived utility of MOOCs in both undergraduate and postgraduate medical education. The results of an online survey demonstrate an openness to, yet in many cases, an unfamiliarity with, this technology in both educators and students. Further exploration of the data reveals a desire for greater levels of guidance in this ever expanding field, and specific curricular areas were highlighted where potential for the use of MOOCs seems particularly enticing. These insights may be helpful in the ongoing debate as to the direction of the field as educators seek to maximise the effectiveness of a blended approach to medical education.


Late results after splenectomy in adult idiopathic thrombocytopenic purpura.
We performed a retrospective study on patients with idiopathic thrombocytopenic purpura (ITP) to evaluate the response to splenectomy in relation to preoperative platelet count. Two groups of patients operated on with laparoscopic or open splenectomy for ITP, with a platelet count ≤30,000/μL (study group: 22 patients) and >30,000/μL (control group: 18 patients), respectively, were compared. The two groups were homogeneous in relation to age, sex, length of preoperative steroid therapy, and time interval between diagnosis and surgery (Student t test with P > .1). The results of surgery were evaluated at one year after splenectomy. Positive response to surgery, according to the American Society of Hematologic Guidelines, was considered in patients with a postoperative platelet count ≥100,000/μL or in patients with a postoperative platelet count ≥30,000/μL and a twofold increase in platelet count from baseline, in the absence of bleeding. The postoperative platelet count increase rate was statistically related to preoperative platelet count in both the study and control groups. Statistical analysis was performed using the Student's t test for independent sample and the Pearson correlation in a 2-tailed test. No relationship between preoperative platelet count and postoperative platelet percent increase was observed in the control group (r = -0.41; P = .089), whereas a significant negative correlation (r = -0.68; P = .0004) was found in the study group. A higher increase of postoperative percent platelet count may be predicted in patients with a low preoperative platelet count.


Genetic screening for PRA-associated mutations in multiple dog breeds shows that PRA is heterogeneous within and between breeds.
To assess the extent of progressive retinal atrophy (PRA) genetic heterogeneity within and between domestic dog breeds. DNA from 231 dogs with PRA, representing 36 breeds, was screened for 17 mutations previously associated with PRA in at least one breed of dog. Screening methods included amplified fragment size discrimination using gel electrophoresis or detection of fluorescence, (TaqMan(®) ; Life Technologies, Carlsbad, CA, USA) allelic discrimination, and Sanger sequencing. Of the 231 dogs screened, 129 were homozygous for a PRA-associated mutation, 29 dogs were carriers, and 73 were homozygous for the wild-type allele at all loci tested. In two of the 129 dogs, homozygous mutations were identified that had not previously been observed in the respective breeds: one Chinese Crested dog was homozygous for the RCD3-associated mutation usually found in the Cardigan Welsh Corgi, and one Standard Poodle was homozygous for the RCD4-associated mutation previously reported to segregate in Gordon and Irish Setters. In the majority of the breeds (15/21) in which a PRA-associated mutation is known to segregate, cases were identified that did not carry any of the known PRA-associated mutations. Progressive retinal atrophy in the dog displays significant genetic heterogeneity within as well as between breeds. There are also several instances where PRA-associated mutations segregate among breeds with no known close ancestry.


Resist shaping for replication of micro-optical elements with continuous relief in fused silica.
To improve the replicating quality in producing micro-optical elements with continuous relief, the resist layer on substrate is shaped against the negative stamp by dry etching to enable it to have the same geometry as the pattern area of the negative stamp. The negative stamp is then aligned with and imprinted into the shaped resist. The produced continuous relief is transferred into the substrate by dry etching as well. Experiment results indicate that this method has both the long service life of a negative stamp and easy filling of a positive stamp.


Auramine O interaction with DNA: a combined spectroscopic and TD-DFT analysis.
In this work, the interaction between an Auramine O (AuO) fluorescent molecular rotor and natural DNA, its thermodynamic aspects and the resulting variation of the optical properties upon binding are addressed by a combined spectroscopic (UV-vis and fluorescence) and computational approach. DNA binding causes a shift in the maximum of absorption from 432 nm to 444 nm, a decrease of the extinction coefficient and a dramatic enhancement of fluorescence emission, these results being in agreement with intercalation into the polynucleotide helix. Intercalation is indeed confirmed by the thermodynamic parameters for the binding reaction (in particular, the highly negative ΔH). Theoretical modelling at the TD-DFT level was done on a simplified model system consisting of the AuO molecule intercalated between two DNA base pairs. The evolution of the calculated vertical transitions quantitatively reproduces the experimentally observed hypo- and bathochromic shifts, thus confirming the intercalation hypothesis.


Sterile syringe access conditions and variations in HIV risk among drug injectors in three cities.
Better sterile syringe access should be associated with a lower likelihood of syringe re-use and receptive syringe sharing, although few empirical studies have examined gradients in syringe access using both individual and ecological data. In this study, we compare syringe re-use and receptive syringe sharing among injection drug users (IDUs) with syringe exchange program (SEP) and legal over-the-counter pharmacy access with limits on syringes that can be purchased, exchanged or possessed to IDUs with no pharmacy sales but unlimited syringe access through SEPs. We address three questions: (1) Does residing in an area with no legal syringe possession increase the likelihood of police contact related to possessing drug paraphernalia? (2) Among direct SEP users, is use of more permissive SEPs associated with less likelihood of syringe re-use and receptive syringe sharing? (3) Among non-SEP users, is residing in an area with pharmacy access associated with lower likelihood of syringe re-use and receptive syringe sharing? Quantitative survey of IDUs recruited from SEPs, subject nomination and outreach methods. Multivariate analyses compared police contact, syringe re-use and receptive syringe sharing among IDUs recruited in three cities. In multivariate analyses, we found that police contact was associated independently with residing in the area with no legal possession of syringes; among SEP users, those with access to SEPs without limits had lower syringe re-use but not lower syringe sharing; and that among non-SEP users, no significant differences in injection risk were observed among IDUs with and without pharmacy access to syringes. We found that greater legal access to syringes, if accompanied by limits on the number of syringes that can be exchanged, purchased and possessed, may not have the intended impacts on injection-related infectious disease risk among IDUs.


Early diagnosis of fibrodysplasia ossificans progressiva.
Fibrodysplasia ossificans progressiva is a rare and disabling genetic condition characterized by congenital malformation of the great toes and by progressive heterotopic ossification in specific anatomic patterns. Most patients with fibrodysplasia ossificans progressiva are misdiagnosed early in life before the appearance of heterotopic ossification and undergo diagnostic procedures that can cause lifelong disability. Recently, the genetic cause of fibrodysplasia ossificans progressiva was identified, and definitive genetic testing for fibrodysplasia ossificans progressiva is now available before the appearance of heterotopic ossification. We recently evaluated 7 children for diagnosis of fibrodysplasia ossificans progressiva before the onset of heterotopic ossification. A medical history, physical examination, and skeletal survey were obtained on all of the patients, as well as clinical genetic testing for the canonical fibrodysplasia ossificans progressiva mutation. All 7 of the children (4 girls and 3 boys; ages 3 months to 6 years) had congenital malformations of the great toes, but none had radiographic evidence of heterotopic ossification at the time of evaluation. Five of the 7 children had soft tissue lesions of the neck and back, suggestive of early fibrodysplasia ossificans progressiva flare-ups, 3 of whom had undergone invasive diagnostic procedures that exacerbated their condition. Two children had no history or signs of soft tissue swelling or flare-ups. DNA sequence analysis found that all 7 of the children had the recurrent fibrodysplasia ossificans progressiva missense mutation, a single nucleotide substitution (c.617G>A) at codon 206 in the glycine-serine activation domain of activin receptor IA, a bone morphogenetic protein type 1 receptor. Clinical suspicion of fibrodysplasia ossificans progressiva early in life on the basis of malformed great toes can lead to early clinical diagnosis, confirmatory diagnostic genetic testing, and the avoidance of additional harmful diagnostic and treatment procedures. This is the first report of genetic confirmation of fibrodysplasia ossificans progressiva before the appearance of heterotopic ossification. Pediatricians should be aware of the early diagnostic features of fibrodysplasia ossificans progressiva, even before the appearance of heterotopic ossification. This awareness should prompt early genetic consultation and testing and the institution of assiduous precautions to prevent iatrogenic harm.


Molecular characterization of group A rotaviruses in Mukuru slums Kenya: detection of novel strains circulating in children below 5 years of age.
Gastroenteritis is a public health concern due to high morbidity and mortality among children. Rotaviruses are the leading etiological agents of severe gastroenteritis in children and accounts for more than half a million deaths per year in Africa. The study aimed at investigating the rotavirus genotypes that were circulating in children aged 5 years and below in and around Mukuru slums in Nairobi County Kenya. A purposive cross sectional sampling method was applied where 166 samples were collected from children below 5 years of age and taken to Kenya Medical Research Institute virology laboratory. Presence of rotaviruses was determined using reverse transcription polymerase chain reaction, while extraction was done using ZR Soil/Fecal RNA MicroPrep™ extraction kit. This was followed by reverse transcription and genotyping using various group A rotavirus primers. The G type was successfully determined in 37 (92.5%), while the P type was successfully determined in 35 (87.5%) of the 40 (24%) page positive samples. Type G1 was the most predominant of the G types (40.5%), and the incidences of G3 and G9 were 21.6 and 32.4% respectively. Mixed types G3/G9 were detected at 5.4%. Three P types existed in Mukuru slums, P[8] (60%), P[6] (22.9%), P[4] (11.4) and their relative incidence varied over the 15 months of this study. The G types and P types detected in this study are important causes of acute gastroenteritis in Mukuru slums Nairobi Kenya. An indication that the prevalence of certain genotypes may change over a rotavirus season is significant and mirrors observations from studies in other tropical climates. Thus monitoring of the genotypic changes among circulating viruses should be encouraged over the coming years.


A multiscale modelling approach to assess the impact of metabolic zonation and microperfusion on the hepatic carbohydrate metabolism.
The capacity of the liver to convert the metabolic input received from the incoming portal and arterial blood into the metabolic output of the outgoing venous blood has three major determinants: The intra-hepatic blood flow, the transport of metabolites between blood vessels (sinusoids) and hepatocytes and the metabolic capacity of hepatocytes. These determinants are not constant across the organ: Even in the normal organ, but much more pronounced in the fibrotic and cirrhotic liver, regional variability of the capillary blood pressure, tissue architecture and the expression level of metabolic enzymes (zonation) have been reported. Understanding how this variability may affect the regional metabolic capacity of the liver is important for the interpretation of functional liver tests and planning of pharmacological and surgical interventions. Here we present a mathematical model of the sinusoidal tissue unit (STU) that is composed of a single sinusoid surrounded by the space of Disse and a monolayer of hepatocytes. The total metabolic output of the liver (arterio-venous glucose difference) is obtained by integration across the metabolic output of a representative number of STUs. Application of the model to the hepatic glucose metabolism provided the following insights: (i) At portal glucose concentrations between 6-8 mM, an intra-sinusoidal glucose cycle may occur which is constituted by glucose producing periportal hepatocytes and glucose consuming pericentral hepatocytes, (ii) Regional variability of hepatic blood flow is higher than the corresponding regional variability of the metabolic output, (iii) a spatially resolved metabolic functiogram of the liver is constructed. Variations of tissue parameters are equally important as variations of enzyme activities for the control of the arterio-venous glucose difference.


Maternal sexuality during first pregnancy and after childbirth.
Information about sexual activity, enjoyment and libido was obtained at intervals from 119 primiparous women during a longitudinal survey of maternal emotional health in pregnancy and for a year after delivery. Most subjects described some reduction in the frequency of sexual intercourse and a diminution of libido and sexual enjoyment during pregnancy; this was most marked in the third trimester. After delivery, about a third of subjects had resumed intercourse by six weeks and nearly everyone had done so by three months. Nevertheless, 77% and 57% of the women were having intercourse less often at three and 12 months after delivery respectively, in comparison with the month before they became pregnant. Selected variables were examined for relationship with a low, or reduced frequency of intercourse and with a lack of enjoyment. Significant associations were found with aspects of maternal personality and childhood relationships, marital conflict, maternal depression, previous miscarriages, difficulties in conceiving and fears of harming the fetus. Nausea and vomiting during pregnancy, the mode of delivery and related obstetric and medical variables, breast-feeding and characteristics of the baby, did not appear to significantly influence maternal sexuality.


Chromosomal location of a gene (nmpA) involved in expression of a major outer membrane protein in Escherichia coli.
The phenotypic expression of protein E, a recently described major outer membrane protein, is associated with a mutation at a locus on the Escherichia coli chromosome that we call nmpA. nmpA is located between rbsK and uncA at 82.7 min on the E. coli linkage map. The nmpA locus is also the site of the mutations which lead to the formation of major outer membrane proteins Ic or e. It is likely proteins E, Ic, and e are closely related or identical. The mutant nmpA allele is dominant.


Evolutionary mechanisms shaping the genetic population structure of marine fishes; lessons from the European flounder (Platichthys flesus L.).
A number of evolutionary mechanisms have been suggested for generating low but significant genetic structuring among marine fish populations. We used nine microsatellite loci and recently developed methods in landscape genetics and coalescence-based estimation of historical gene flow and effective population sizes to assess temporal and spatial dynamics of the population structure in European flounder (Platichthys flesus L.). We collected 1062 flounders from 13 localities in the northeast Atlantic and Baltic Seas and found temporally stable and highly significant genetic differentiation among samples covering a large part of the species' range (global F(ST) = 0.024, P < 0.0001). In addition to historical processes, a number of contemporary acting evolutionary mechanisms were associated with genetic structuring. Physical forces, such as oceanographic and bathymetric barriers, were most likely related with the extreme isolation of the island population at the Faroe Islands. A sharp genetic break was associated with a change in life history from pelagic to benthic spawners in the Baltic Sea. Partial Mantel tests showed that geographical distance per se was not related with genetic structuring among Atlantic and western Baltic Sea samples. Alternative factors, such as dispersal potential and/or environmental gradients, could be important for generating genetic divergence in this region. The results show that the magnitude and scale of structuring generated by a specific mechanism depend critically on its interplay with other evolutionary mechanisms, highlighting the importance of investigating species with wide geographical and ecological distributions to increase our understanding of evolution in the marine environment.


Lasting neuron depression induced by high potassium and its prevention by low calcium and NMDA receptor blockade.
Spreading depression-like neuron depolarization was induced in CA1 of hippocampal tissue slices by irrigation with artificial cerebrospinal fluid containing 133.5 mM K+ for 8-40 min. Evoked responses disappeared during irrigation with high-K+ solution. Following 8-20 min irrigation orthodromic responses showed a triphasic recovery cycle: early partial return with evidence of neuron hyperexcitability, then secondary depression and finally slow partial recovery. After 30 min or more of high-K+ exposure, ortho- and antidromic responses remained severely depressed for at least 5.5 to 6.5 h. When, however, the tissue was deprived of calcium, or N-methyl-D-aspartate (NMDA) receptors were blocked by 10 microM 3-((+-)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP), then evoked responses recovered partially after a 30- or 40-min high-K+ exposure. Post-exposure hyperexcitability was not prevented by CPP. We conclude that prolonged depolarization by elevated K+ causes irreversible neuron damage, which is triggered or accelerated by influx of calcium ions into neurons, mediated in part by NMDA receptor activation.


The Problem of Occult Pyelonephritis: Acute Pyelonephritis in Patients Presenting Only with Lower Urinary Tract Symptoms.
Pyelonephritis is a prevalent diagnosis of emergency department visits. It commonly presents as flank pain and costovertebral tenderness with urinary symptoms. However, some cases occur without typical symptoms. Our study highlights the dubious presentation of pyelonephritis with lower urinary tract symptoms (LUTS) only. This study was conducted at a tertiary care hospital, where charts and files were reviewed from January 11, 2018 to February 28, 2019 for all the patients with a diagnosis of acute pyelonephritis from medical records. In our study, 521 patients were included and 492 (94%) of the participants were suffering from pyelonephritis. Approximately 22.8% of the patients showed the absence of both flank pain and costovertebral tenderness but were diagnosed with pyelonephritis based on computed tomography (CT) and magnetic resonance imaging (MRI). Moreover, 27% of the patients reported upper urinary tract symptoms only and were diagnosed by CT or MRI findings. Out of that only 24% and 16% of the patients reported flank pain and costovertebral tenderness, respectively. Insignificant associations with pyelonephritis were found for age, gender and other comorbidities. Our study showed a significant number of patients with pyelonephritis without any upper urinary tract symptoms. Patients with LUTS should be evaluated further by imaging if they belong to the high-risk population.


Exploring the role of striatal D1 and D2 medium spiny neurons in action selection using a virtual robotic framework.
The basal ganglia have been hypothesized to be involved in action selection, i.e. resolving competition between simultaneously activated motor programs. It has been shown that the direct pathway facilitates action execution whereas the indirect pathway inhibits it. However, as the pathways are both active during an action, it remains unclear whether their role is co-operative or competitive. In order to investigate this issue, we developed a striatal model consisting of D1 and D2 medium spiny neurons (MSNs) and interfaced it to a simulated robot moving in an environment. We demonstrate that this model is able to reproduce key behavioral features of several experiments involving optogenetic manipulation of the striatum, such as freezing and ambulation. We then investigate the interaction of D1- and D2-MSNs. We find that their fundamental relationship is co-operative within a channel and competitive between channels; this turns out to be crucial for action selection. However, individual pairs of D1- and D2-MSNs may exhibit predominantly competition or co-operation depending on their distance, and D1- and D2-MSNs population activity can alternate between co-operation and competition modes during a stimulation. Additionally, our results show that D2-D2 connectivity between channels is necessary for effective resolution of competition; in its absence, a conflict of two motor programs typically results in neither being selected.


Targeting the insulin-like growth factor axis as a cancer therapy.
The insulin-like growth factor (IGF) axis has been implicated in malignant transformation and in tumor cell biology. Human population studies have demonstrated that high levels of circulating IGF-I are associated with an increased risk of certain malignancies. Many model systems show that IGFs stimulate tumor cell proliferation, survival and metastasis. In a new era of anticancer treatments aimed at tumor-specific targets, efforts are in progress for the development of novel anti-IGF therapies. Disrupting type I IGF-receptor (IGF-IR) function in vitro and in vivo results in tumor growth inhibition in several model systems. Antireceptor therapies in particular have provided encouraging results leading to the approval of the first Phase I human clinical trial targeting IGF-IR. Additional methods to decrease levels of circulating IGF-I and II have also been developed. In principle, a benefit of targeted therapies could be their relative lack of toxicity compared with conventional chemotherapy. Anti-IGF-IR therapies, however, raise theoretical concerns for the development of serious side effects, including diabetes. As targeted therapies against the IGF axis continue to be developed, efforts will need to be made to minimize the side effects that result from blocking normal ligand and receptor-induced functions.


[Use of PCR for the identification of representatives of Lactobacillus and Streptococcus].
REP- and ERIC-PCR genome analysis of lactic acid bacteria of genera Lactobacillus and Streptococcus have shown the presence of REP- and ERIC-repetitive sequences with high degree of homology. Amplification products which separation in agar gel results in formation of a specific fingerprint are obtained under REP- and ERIC-PCR. It is shown that REP- and ERIC-specific primers can be used for PCR identification of both enterobacteria and lactic acid bacteria isolated from different ecological niches.


Coordinated series of studies to evaluate characteristics and mechanisms of acute coronary syndromes in high-risk patients randomly assigned to enoxaparin or unfractionated heparin: design and rationale of the SYNERGY Library.
Clinical trials and accompanying substudies in patients with acute coronary syndromes (ACS) have over the last several years yielded a wealth of knowledge about the pathophysiology and management of this high-risk condition. The Superior Yield of the New strategy of Enoxaparin, Revascularization, and GlYcoprotein IIb/IIIa inhibitors (SYNERGY) trial is a large-scale, randomized, controlled trial evaluating the effect of enoxaparin and unfractionated heparin on death and myocardial infarction in high-risk patients presenting with non-ST-segment elevation ACS. The SYNERGY Library has been designed as a coordinated series of investigations with simultaneous data acquisition on the same cohort of approximately 500 SYNERGY patients at 60 centers in North America. Specifically, electrocardiograms, coronary arteriograms, inflammatory markers, coagulation studies, and genetic samples will be collected and processed at core laboratory facilities, and the results will be stored in a central repository. This novel strategy for substudy investigation is unprecedented in cardiovascular clinical trials. The goal is to gain significant understanding about this patient population, discover new principles of pathophysiology, identify novel pharmacologic targets, and streamline further drug development. It is hoped that the SYNERGY Library will serve as a model for future substudy design to maximize academic insight within the framework of a large-scale, multicenter trial.


Epigenetic alterations in human prostate cancers.
Human prostate cancer cells carry a myriad of genome defects, including both genetic and epigenetic alterations. These changes, which can be maintained through mitosis, generate malignant phenotypes capable of selective growth, survival, invasion, and metastasis. During prostatic carcinogenesis, epigenetic changes arise earlier than genetic defects, linking the appearance of epigenetic alterations in some way to disease etiology. The most common genetic defect thus far described, leading to fusion transcripts between the androgen-regulated gene TMPRSS2 and genes from the ETS family of transcription factors, likely endows prostate cancer cells with the ability to co-opt androgen signaling, the major prostate differentiation pathway, to support the malignant phenotype. Whether epigenetic changes promote the appearance of TMPRSS2-ETS family fusion transcripts or collaborate with fusion transcript expression in the pathogenesis of prostate cancer has not been established. However, a growing list of epigenetic alterations has provided new opportunities for clinical tests that might aid in prostate cancer screening, detection, diagnosis, staging, and risk stratification. The epigenetic changes appear to be more attractive than genetic changes as prostate cancer biomarkers because epigenetic alterations are present in a greater fraction of prostate cancer cases than any of the known genetic defects. In addition, an emerging generation of assay strategies for detection of specific DNA sequences carrying (5-me)C, the major epigenetic genome mark, has pushed somatic epigenetic alterations to the forefront of molecular biomarker assay development for cancer. Finally, a growing portfolio of epigenetic drugs, capable of reversing the phenotypic consequences of somatic epigenetic defects, has entered clinical trials for prostate cancer in the search for a new rational therapy for the disease.


The burden of treatment failure in type 2 diabetes.
In type 2 diabetes, therapies to maintain blood glucose control usually fail after several years. We estimated the glycemic burden that accumulates from treatment failure and describe the time course and predictors of failure. A prospective, population-based study using retrospective observational data. We identified all 7208 complete courses of treatment with nondrug therapy, sulfonylurea monotherapy, metformin monotherapy, and combination oral antihyperglycemic therapy between 1994 and 2002, inclusive, among members of the Kaiser Permanente Northwest Region. We calculated mean cumulative glycemic burden, defined as HbA(1c)-months >8.0 or 7.0% for each treatment. We then measured the likelihood that the next HbA(1c) would exceed 8.0 and 7.0% after HbA(1c) exceeded each of ten hypothetical treatment thresholds. Finally, we estimated multivariate logistic regression models to predict when HbA(1c) would continue to deteriorate. In this well-controlled population, the average patient accumulated nearly 5 HbA(1c)-years of excess glycemic burden >8.0% from diagnosis until starting insulin and about 10 HbA(1c)-years of burden >7.0%. Whenever patients crossed the American Diabetes Association-recommended treatment threshold of 8.0%, their next HbA(1c) result was as likely to be <8.0 as >8.0%. Multivariate prediction models had highly statistically significant coefficients, but predicted <10% of the variation in future HbA(1c) results. Clinicians should change glucose-lowering treatments in type 2 diabetes much sooner or use treatments that are less likely to fail. An action point at 7.0% or lower is more likely to prevent additional deterioration than the traditional action point of 8.0%.


Sleep Deprivation Disrupts Recall of Conditioned Fear Extinction.
Learned fear is crucial in the development and maintenance of posttraumatic stress disorder (PTSD) and other anxiety disorders, and extinction of learned fear is necessary for response to exposure-based treatments. In humans, research suggests disrupted sleep impairs consolidation of extinction, though no studies have examined this experimentally using total sleep deprivation. Seventy-one healthy controls underwent a paradigm to acquire conditioned fear to a visual cue. Twenty-four hours after fear conditioning, participants underwent extinction learning. Twenty-four hours after extinction learning, participants underwent extinction recall. Participants were randomized to three groups: 1) well-rested throughout testing ("normal sleep"; n = 21); 2) 36 hours total sleep deprivation before extinction learning ("pre-extinction deprivation"; n = 25); or 3) 36 hours total sleep deprivation after extinction learning and before extinction recall ("post-extinction deprivation"; n = 25). The groups were compared on blink EMG reactivity to the condition stimulus during extinction learning and recall. There were no differences among the three groups during extinction learning. During extinction recall, the pre-extinction deprivation group demonstrated significantly less extinction recall than the normal sleep group. There was no significant difference between the normal sleep and post-extinction deprivation group during extinction recall. Results indicated sleep deprivation prior to extinction training significantly disrupts extinction recall. These findings suggest that (1) sleep deprivation in the immediate aftermath of trauma could be a potential contributor to PTSD development and maintenance via interference with natural extinction processes and (2) management of sleep symptoms should be considered during extinction-based therapy.


Prevalence of hypertension and obesity in patients with type 2 diabetes mellitus in observational studies: a systematic literature review.
Hypertension and obesity are known to contribute, directly or indirectly, to the development of long-term complications of type 2 diabetes mellitus (T2DM). Knowing the prevalence of these comorbidities is important for determining the size of the population that may benefit from strategies that reduce blood pressure and weight while controlling blood glucose. In this systematic literature review, electronic searches of PubMed, Embase, and the Cochrane Library were conducted to identify observational studies of hypertension and/or obesity prevalence in patients with T2DM throughout the world. The searches were limited to studies reported in English from January 1, 2001 to February 16, 2012. From a total of 2,688 studies, 92 observational studies provided prevalence rates for hypertension and/or obesity specifically in adults with T2DM. Fifteen studies of specific subtypes of hypertension or subpopulations with T2DM were subsequently excluded, leaving 78 studies (in 77 articles) for inclusion in this article. Of these, 61studies reported hypertension prevalence, 44 reported obesity prevalence, and 12 reported the prevalence of hypertension with obesity. Most studies had a low risk of bias regarding diagnosis of T2DM (70/78), hypertension (59/69), or obesity (45/47). The continental regions with the most observational studies of hypertension or obesity prevalence were Europe (n = 30) and Asia (n = 26). Hypertension rates typically were high in all regions; most studies presented rates above 50%, and many presented rates above 75%. Obesity rates exceeded 30% in 38 of 44 studies and 50% in 14 of 44 studies, especially those assessing central obesity (based on waist circumference). Among obese adults, hypertension rates were at or above 70% in Asia and above 80% in Europe; rates were lower in North and South America but still above 30%. Around the world, hypertension and obesity, separately or together, are common comorbidities in adults with T2DM.


The peculiar challenges of blindness prevention in Nigeria: a review article.
To describe the challenges peculiar to Nigeria in the implementation of vision 2020: the right to sight and to proffer solutions as to the way forward A review of the recently completed national blindness survey, current literature, and the advocacy experience of the Ophthalmological Society of Nigeria. The prevalence of blindness in Nigeria is 0.78%. Over 43% of these are blind from cataracts and a further 9% from uncorrected aphakia or complications of couching. 50% of all cataract interventions are carried out by itinerant couchers. Other major causes of blindness are glaucoma (16%) corneal opacities (12%), trachoma (4%), optic atrophy (3%) onchocerciasis (1%) and macular disease (1%). 70% of these are either preventable or reversible. Nigeria has a relatively favourable ophthalmologist/population ratio of about 2.8 per million, but has a low Cataract surgical rate of 300 per million per year. The reasons for this include a lack of ownership of blindness prevention programs, a lack of political will and parlous state of funding for vision 2020. There is an abdication of responsibility for both training and services on the part of government to the International NonGovernmental organisations. Teaching hospitals no longer generate enough patient surgical load to support training. We estimate it would cost N8.5 billion ($56.8 million) to sustain the WHO recommended Cataract Surgical Rate of 3000 per million per year in Nigeria. Nigeria is not headed in the direction of meeting Vision 2020 targets. Advocacy involving funding through the MDGs, needs to be intensified.


Prediction of daily and lactation yields of milk, fat, and protein using an autoregressive repeatability test day model.
We evaluated the accuracy of an autoregressive multiple-lactation test day (ATD) model to predict missing test day yields of milk, fat, and protein to obtain cumulative 305-d records for cows with incomplete or in-progress lactations. The data consisted of more than one million observations of daily yields on test days in the first 3 lactations of over 75,000 Portuguese Holstein cows. Differences between actual (estimates from complete lactations using the test interval method) and ATD-predicted 305-d yields were negligible and smaller than those predicted by the test interval method. The ATD procedure tended to slightly underestimate cumulative lactation yields, whereas the test interval method substantially overestimated them. Smaller differences obtained by the ATD procedure resulted in less biased estimates of lactation yield, which also implies greater accuracy. As expected, the correlations between actual and predicted lactation yields increased with the number of test days from 0.831 to 0.997. Average correlations (by parity) between actual and ATD-predicted yields ranged from 0.977 to 0.984. Correlations between actual test day yields and corresponding predicted yields exceeded 0.5 for up to 7 time-intervals from the last test day yield used to predict cumulative yield of projected lactations. These correlations indicate the good predictive ability of the ATD method. From a producer's viewpoint, these advantages underwrite management because most on-farm selection decisions are based on the producing abilities of cows. Implementation of ATD methodology does not require special computing capability and is easily transferable to the farm level.


Initial cyclosporin A but not glucocorticoid treatment promotes recovery of striatal dopamine concentration in 6-hydroxydopamine lesioned mice.
We examined the effects of cyclosporin A (CsA) and glucocorticoid (GC; hydrocortisone sodium succinate) in a mouse model of experimental parkisonism. GC or CsA was administered 30 or 60 min, respectively, prior to intracerebroventricular injection of 6-hydroxydopamine, followed by injection of a similar dose of each drug 3 h later. CsA reduced the extent of depletion of striatal concentrations of dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) associated with dopaminergic neuronal degeneration. GC reduced the extent of homovanillic acid (HVA) depletion in the same region. A combination treatment with CsA and GC did not produce a further enhancement of the recovery of striatal concentrations of monoamines observed with CsA only. Our findings demonstrated the beneficial effects of initial CsA treatment in experimental models of parkinsonism and further support the usefulness of CsA in the treatment of Parkinson's disease.


Bradycardia following carotid endarterectomy.
This study was performed to determine whether bradycardia complicates the postoperative course of patients undergoing carotid endarterectomy (CEA). The records of 216 patients undergoing 233 CEAs over a 2-year period were reviewed. Patients were divided into two groups based on their lowest Surgical Intensive Care Unit (SICU) heart rate (HR). Those with HR < 60 were in the Bradycardic (BRADY) group and those with HR > or = 60 were in the Non-Bradycardic (NON-BRADY) group. One hundred and sixteen patients developed bradycardia, with a mean (+/- SEM) HR of 51.1 +/- 0.5, compared with 117 NON-BRADY patients with a mean HR of 70.6 +/- 0.9 (P < 0.0005). There were no significant differences between the groups in age, use of cardioactive drugs, SICU severity of illness, or length of SICU stay. The systolic blood pressure for BRADY patients averaged 144 +/- 2.2 on admission and 144 +/- 2.2 (P = NS) in the SICU, while that of NON-BRADY patients rose from 143 +/- 2.3 on admission to 156 +/- 2.5 (P = 0.001). Fifty-four patients receiving a second CEA had a SICU HR not significantly different from those patients undergoing a first CEA. Of 17 patients who underwent bilateral CEAs during the study period, SICU HRs averaged 65.1 +/- 3.7 after the first procedure and 64.7 +/- 3.6 after the second (P = NS). The authors conclude that bradycardia following CEA is a frequent but benign postoperative finding that does not affect outcome, cause significant hypotension, or prolong the SICU stay.


HIV infection of astrocytes compromises inter-organelle interactions and inositol phosphate metabolism: A potential mechanism of bystander damage and viral reservoir survival.
HIV-associated neurological dysfunction is observed in more than half of the HIV-infected population, even in the current antiretroviral era. The mechanisms by which HIV mediates CNS dysfunction are not well understood but have been associated with the presence of long-lasting HIV reservoirs. In the CNS, macrophage/microglia and a small population of astrocytes harbor the virus. However, the low number of HIV-infected cells does not correlate with the high degree of damage, suggesting that mechanisms of damage amplification may be involved. Here, we demonstrate that the survival mechanism of HIV-infected cells and the apoptosis of surrounding uninfected cells is regulated by inter-organelle interactions among the mitochondria/Golgi/endoplasmic reticulum system and the associated signaling mediated by IP3 and calcium. We identified that latently HIV-infected astrocytes had elevated intracellular levels of IP3, a master regulator second messenger, which diffuses via gap junctions into neighboring uninfected astrocytes resulting in their apoptosis. In addition, using laser capture microdissection, we confirmed that bystander apoptosis of uninfected astrocytes and the survival of HIV-infected astrocytes were dependent on mitochondrial function, intracellular calcium, and IP3 signaling. Blocking gap junction channels did not prevent an increase in IP3 or inter-organelle dysfunction in HIV-infected cells but reduced the amplification of apoptosis into uninfected neighboring cells. Our data provide a mechanistic explanation for bystander damage induced by surviving infected cells that serve as viral reservoirs and provide potential targets for interventions to reduce the devastating consequences of HIV within the brain.


Understanding the Perceived Threat of the Risk of Graft Rejections: A Middle-Range Theory.
From a clinical viewpoint, graft rejection is one of the greatest threats faced by an organ transplant recipient (OTR). We propose a middle-range theory (MRT) of Perceived Threat of the Risk of Graft Rejection (PTRGR) as a contribution to the practice of transplant nursing. It could also apply to the detection of risky protective behavior, that is, isolation, avoidance, or non-adherence. The proposed MRT covers the following concepts and the relationship between them: transplant care needs, threat reducing interventions, intervening variables, level of PTRGR, protective strategies, and evidence-based practice. Parts of this theory have been empirically tested and support the suggested relationship between some of the concepts. Further tests are needed to strengthen the theoretical links. The conceptual framework might serve as a guide for transplant nurses in their efforts to promote post-transplant health and reduce threat-induced emotions.


The persistence of emamectin benzoate in marine sediments with different organic matter regimes, temperature conditions, and antibiotic presence.
The objectives of this 318-day study are to determine half-lives of the anti-sea lice medication emamectin benzoate (EMB) under conditions present in sediments at aquaculture sites and document the degradation of EMB into its main metabolite desmethyl emamectin benzoate (DES). Tested conditions include different matrix types (sand, mud), two temperatures (4, 10 degrees), organic matter presence (fish feed waste and feces), and the presence of oxytetracycline. We document a transformation ratio of EMB to DES of 0.16 to 4.4 % and show that the co-presence of oxytetracycline increases EMB calculated half-lives to values >6000 days for mud matrices. EMB incubated in high organic enrichment regimes was not observed to degrade at 4 degrees. Multivariate analyses show interactions between sediment conditions (matrix, temperature, organic matter [OM], oxytetracycline) influence EMB persistence and DES:EMB ratios. Ranges of EMB half-lives and information on metabolites can be used to anticipate potential effects on marine communities.


Okadaic acid inhibits activation of K-Cl cotransport in red blood cells containing hemoglobins S and C.
The sensitivity of red blood cells containing hemoglobins S and C to activation of K-Cl cotransport by osmotic swelling and acidification was reduced by okadaic acid, a specific protein phosphatase inhibitor. The dose-response curve for okadaic acid suggests its action is on a type 1 protein phosphatase. Okadaic acid has been previously shown to inhibit swelling-induced activation of K-Cl cotransport in red blood cells from rabbits, normal humans, and dogs. The present work confirms the observation that okadaic acid blunts the stimulation of K-Cl cotransport by cell swelling. The new information is that okadaic acid reduces the effects of hemoglobins S and C on the volume and pH sensitivity of K-Cl cotransport. Thus the influences of cell volume, pH, and mutant hemoglobins may all be mediated via a common mechanisms that affects the phosphorylation state, either of the K-Cl. cotransporter itself or of a protein that regulates its function.


Severe cerebral white matter involvement in a case of dentatorubropallidoluysian atrophy studied at autopsy.
The pathophysiology of white matter involvement in dentatorubropallidoluysian atrophy (DRPLA) is controversial. Moreover, the clinical repercussions and evolution of these lesions have not been well documented. To describe a case of DRPLA with severe cerebellar white matter involvement. Case report. Patient A 62-year-old woman with DRPLA. When the genetic diagnosis was made, the patient manifested severe ataxia, slight dysarthria, and subcortical cognitive impairment. Cranial magnetic resonance imaging showed atrophy of the cerebellum and brainstem and moderate high-intensity signal alterations in the periventricular cerebral white matter in T2-weighted sequences. In the following 5 years, she developed uncontrolled head movements associated with severe bruxism and tetraparesis, and became deeply demented. New magnetic resonance imaging showed severe diffuse cerebral white matter alterations in T2 sequences with only slight progression of brainstem and cerebellar atrophy. After her death at 67 years of age, the autopsy study showed diffuse myelin pallor, axonal preservation, and reactive astrogliosis in the cerebral white matter, with only mild atherosclerotic changes, and moderate neuronal loss in the cerebellum and brainstem. Leukoencephalopathy could be a prominent finding in some patients with DRPLA, explaining, at least in part, their clinical evolution. In our case, the disproportion between the severity of white matter damage and vascular changes does not support a cardinal role for ischemic mechanisms in leukoencephalopathy.


Aluminium transport in blood serum. Binding of aluminium by human transferrin in the presence of human albumin and citrate.
The binding of Al3+ by human serum transferrin has been investigated by u.v.-visible difference spectroscopy. In the presence of 25 mM-HCO3- at pH 7.4, the apparent association constants were found to be 1.69 x 10(12) M-1 and 5.36 x 10(11) M-1. These association constants are pH-dependent, reducing with both increasing and decreasing pH. The apparent pKa values were found to be 6.7 and 8.2. Competitive assays of binding of Al3+ to transferrin in the presence of citrate and human serum albumin at molar ratios corresponding to those found in normal plasma showed that a considerable amount of Al3+ was not bound to transferrin. Taking a concentration of 5 microM as a typical value observed for the plasma of patients on haemodialysis [Harris & Sheldon (1990) Inorg. Chem. 29, 119-124] the competitive binding assay indicate that approximately 60% of it is bound to transferrin, approximately 34% to albumin and the remainder to citrate. These results therefore suggest that, although transferrin at pH 7.4 is the major Al(3+)-binding component of plasma, an appreciable amount of Al3+ present in patients on haemodialysis may be bound to albumin.


Humidity-dependent reversible aggregation of rhodamine 6G dye immobilized within layered niobate K4Nb6O17.
The spectroscopic behavior of rhodamine 6G (R6G) dye intercalated in layered hexaniobate K4Nb6O17 was investigated. R6G cations were intercalated into the niobate through displacement of preintercalated alkylammonium ions. Powder X-ray diffraction and elemental analysis indicated that the dye molecules were densely accommodated in the interlayer spaces of niobate. The spectroscopic behavior of intercalated R6G was characterized by humidity-dependent aggregation at room temperature. The dye molecules were present dominantly as monomers under humid conditions (93% relative humidity (RH)), while they formed dimers under relatively dry conditions (20% RH). The aggregation-deaggregation of dye occurred reversibly depending on the humidity. The reversible aggregation was not accompanied by a large alteration of the interlayer structure of the sample, because only a small amount of water was adsorbed/desorbed with a small change in the basal spacing of the intercalation compound during the humidity change.


Integrating Machine Learning with Biomedical Signal Processing and Systems Analysis: An Applications-based Course.
The growing importance of data analytics in biomedicine is increasingly becoming recognized in biomedical engineering curricula through the introduction of machine learning classes that generally run in parallel to, but separately from, more traditional engineering courses, such as signal and systems analysis. We propose a new approach that systematically integrates signal processing and systems analysis with key techniques in machine learning. In the proposed course, the student obtains hands-on experience in applying algorithms that can be applied to practical problems of physiological signal conditioning, analysis and interpretation. This is achieved by exposing the student to a sequence of 4 applications-based modules that represent different biomedical engineering problems: human activity recognition from wearable devices, epileptic seizure detection, quantification of dynamic respiratory-cardiac coupling in humans under different conditions, and detection of sleep apnea episodes from heart rate variability data. Within each module, the student gains the experience of working with the data in question "from the ground up". We also introduce a general plan for assessment of student learning, and discuss the expected outcomes and limitations of this integrative approach to teaching.Clinical Relevance- The proposed course is targeted at biomedical engineering students at the senior undergraduate or first-year graduate level who are interested in learning how to analyze physiological signals. The course would also be suitable for clinician-scientists who have prior training in statistics with some exposure to engineering mathematics.


[Regulation of respiration and pulmonary blood circulation in experimental pneumonia].
The experiments on anesthetized cats were carried out to determine the functional state of respiration centre (impulse activity of genuine respiration neurons and electrical activity of major and additional respiration muscles). Functional peculiarities of macro- and microcirculation of the lung tissue were also studied in the open and closed chest in normal subjects and in conditions of experimental pneumonia.


Relating Redox Properties of Polyvinylamine-g-TEMPO/Laccase Hydrogel Complexes to Cellulose Oxidation.
The structure and electrochemical properties of adsorbed complexes based on mixtures of polyvinylamine-g-TEMPO (PVAm-T) and laccase were related to the ability of the adsorbed complexes to oxidize cellulose. PVAm-T10 with 10% of the amines bearing TEMPO moieties (i.e., DS = 10%), adsorbed onto gold sulfonate EQCM-D sensor surfaces giving a hydrogel film that was 7 nm thick, 89% water, and encasing laccase (200 mM) and TEMPO moieties (33 mM). For DS values >10%, all of the TEMPOs in the hydrogel film were redox-active in that they could be oxidized by the electrode. With hydrogel layers made with lower-DS PVAm-Ts, only about half of the TEMPOs were redox-active; 10% DS appears to be a percolation threshold for complete TEMPO-to-TEMPO electron transport. In parallel experiments with hydrogel complexes adsorbed onto regenerated cellulose films, the aldehyde concentrations increased monotonically with the density of redox-active TEMPO moieties in the adsorbed hydrogel. The maximum density of aldehydes was 0.24 μmol/m2, about 10 times less than the theoretical concentration of primary hydroxyl groups exposed on crystalline cellulose surfaces. Previous work showed that PVAm-T/laccase complexes are effective adhesives between wet cellulose surfaces when the DS is >10%. This work supports the explanation that TEMPO-to-TEMPO electron transport is required for the generation of aldehydes necessary for wet adhesion to PVAm.


Knowledge on Bone Banking among Participants in an Orthopaedic Conference: A Preliminary Survey.
Despite increasing use of bone graft in Malaysia, there was still lack of data to quantify knowledge level on bone banking among orthopaedic community who are involved in transplantation related work. Therefore, a survey on awareness in tissue banking specifically bone banking, usage and choice of bone grafts was conducted. From 80 respondents, 82.5% were aware about tissue banking however only 12.5% knew of the existence of tissue banks in Malaysia. Femoral head was the bone allograft most often used as a substitute to autograft. Only 34.8% respondents preferred irradiated bone grafts whilst 46.9% preferred nonirradiated, indicating the need to educate the importance of radiation for sterilising tissues. Exhibition was the most preferred medium for awareness programme to disseminate information about bone banking in the orthopaedic community. The professional awareness is necessary to increase the knowledge on the use of bone graft, hence to increase bone transplantation for musculoskeletal surgeries in the country.


Prescription History Before Opioid Overdose Death: PDMP Data and Responsible Prescribing.
As the opioid epidemic continues, state legislatures and clinicians increasingly utilize Prescription Drug Monitoring Programs (PDMPs). These programs record dates prescribed and filled for all controlled substances, attempting to identify high-risk prescribing. The aims of this study were to (i) examine data from individuals who died of accidental opioid overdose and (ii) compare differences between those with prescriptions documented in Kentucky's PDMP with individuals without recorded prescriptions. This was a retrospective, observational cohort study conducted in Jefferson County, Kentucky. We reviewed records for all opioid overdose death subjects from 2017 and 2018, cross-referencing with prescriptions in Kentucky's PDMP (Kentucky All Schedule Prescription Electronic Reporting System [KASPER]) back to 2014. We performed χ2 analyses for categorical variable comparisons and a separate univariate analysis for age. Of the 575 individuals who died of accidental opioid overdose in Jefferson County during the study period, 379 (65.9%) had prescriptions documented in KASPER. Individuals had a high prevalence of fentanyl on postmortem toxicology. Only one individual had postmortem toxicology positive for buprenorphine, a medication for opioid use disorder (MOUD). Several subjects experienced what we termed see-saw MOUD prescribing (prescriptions alternating between MOUD and other controlled substances including full agonists), and multiple prescriptions were apparently written and/or filled for deceased subjects. Review of PDMP data in deceased patients can prevent unnecessary opioid prescribing and optimize clinical practice. Buprenorphine may have a protective effect in opioid dependence, but access must be consistent. Providers should be aware of see-saw MOUD prescribing and understand the effects on patient care. In response to the prescriptions filled for deceased individuals, legislators could enact a policy such as Void All Prescriptions or VAP alerts to cancel all prescriptions for individuals who have died, reducing drug diversion. It is vital that providers routinely use PDMP data along with counseling and other treatment strategies to optimize patient care.


Attenuated inspiratory muscle metaboreflex in endurance-trained individuals.
The inspiratory metaboreflex is activated during loaded breathing to task failure and induces sympathetic activation and peripheral vasoconstriction that may limit exercise performance. Inspiratory muscle training appears to attenuate the inspiratory metaboreflex in healthy subjects. Since whole body aerobic exercise training improves breathing endurance and inspiratory muscle strength, we hypothesized that endurance-trained individuals would demonstrate a blunted inspiratory muscle metaboreflex in comparison to sedentary individuals. We studied 9 runners (23±0.7 years; maximal oxygen uptake [VO2 max] = 53 ± 4 ml kg(-1) min(-1)) and 9 sedentary healthy volunteers (24±0.7 years; VO2 max = 37 ±2 ml kg(-1) min(-1)). The inspiratory muscle metaboreflex was induced by breathing against an inspiratory load of 60% of maximal inspiratory pressure (MIP), with prolonged duty cycle. Arterial pressure, popliteal blood flow, and heart rate were measured throughout the protocol. Loaded breathing to task failure increased mean arterial pressure in both sedentary and endurance-trained individuals (96±3 to 100±4 mmHg and 101±3 to 110±5 mmHg). Popliteal blood flow decreased in sedentary but not in trained individuals (0.179±0.01 to 0.141±0.01 cm/s, and 0.211±0.02 to 0.214±0.02 cm/s). Similarly, popliteal vascular resistance increased in sedentary but not in trained individuals (559±35 to 757±56 mmHg s/cm, and 528±69 to 558±64 mmHg s/cm). These data demonstrate that endurance-trained individuals have an attenuated inspiratory muscle metaboreflex.


Paravertebral blockade with propofol sedation versus general anesthesia for elective endovascular abdominal aortic aneurysm repair.
The objective of this study was to evaluate the applicability of paravertebral blockade (PVB) for endovascular abdominal aortic aneurysm repair compared with general anesthesia (GA). Data from patients who underwent elective infrarenal endovascular abdominal aortic aneurysm repair between August 2001 and July 2002 using PVB or GA were retrospectively reviewed and compared with respect to risk factors, intraoperative hemodynamic characteristics, operative outcome, and complications. Ten patients underwent elective infrarenal endovascular abdominal aortic aneurysm repair under PVB, whereas 15 patients were operated on under GA. One conversion from PVB to GA was necessary for block failure. The perioperative (< 30 days) cardiovascular morbidity and overall mortality were zero in both groups. The PVB group benefited significantly with respect to the incidence of intraoperative hypotension (p < .05) and blood pressure lability (p < .01), as well as postoperative nausea (p < .01). Our preliminary results indicate that PVB is feasible and can be performed safely in a patient population with significant comorbidities.


Evaluating the Implementation of Ontario's Organ and Tissue Donation Physician Leadership Model: Mapping a Way Forward.
The demand for solid organ transplantation has spurred countries around the world to search for innovative policies and practices to increase the supply of organs. Spain has become a global reference point for organ donation with the highest transplantation rates. In Ontario, Canada the Ontario Trillium Gift of Life (TGLN) has sought to replicate some of the successes in Spain. In particular, TGLN's implementation of the Physician Leadership Model has been viewed as a promising strategy to improve donation conversion rates. The objective of this study was to evaluate the implementation of TGLNs (TGLN) Physician Leadership Model by examining critical implementation process variables (education/training, communication, satisfaction, participation and reach). This mixed-method implementation evaluation included data from all members of the Physician Leadership Model including the Chief Medical Officer, five Regional Medical Leads (RMLs), and the 52 Hospital Donation Physicians (HDPs). Social Network Analysis (SNA) surveys were sent to all 52 HDPs and yielded an 85% rate. Analysis included constructing sociograms and qualitatively analyzing interviews. TGLN's PLM was poised for success by utilizing the existing RMLs' network as a foundation. The social network analysis measures, particularly participation and reach, indicated the PLM was quite dense (ie, the degree to which members are connected) at baseline. HDPs reported communication to be facilitated by their connections to their RMLs. Early evaluative data indicated that lack of education and training was viewed by HDPs as a barrier, and thus more capacity would need to be directed to this issue. Overall, HDPs reported that various intended outcomes were being met. We have demonstrated that an implementation evaluation helps us to understand which elements of the PLM were successful and which elements required immediate attention. This evaluation helped to highlight the successes and challenges in implementing the TGLN Physician Leadership Model in Ontario. Social network analysis of publicly funded capacity building systems has been identified as a promising area for health program evaluation to answer questions at a system level, such as identifying service provisions among information exchange networks and ultimately better health care.


Targeted metabolomics reveals a male pheromone and sex-specific ascaroside biosynthesis in Caenorhabditis elegans.
In the model organism Caenorhabditis elegans, a class of small molecule signals called ascarosides regulate development, mating, and social behaviors. Ascaroside production has been studied in the predominant sex, the hermaphrodite, but not in males, which account for less than 1% of wild-type worms grown under typical laboratory conditions. Using HPLC-MS-based targeted metabolomics, we show that males also produce ascarosides and that their ascaroside profile differs markedly from that of hermaphrodites. Whereas hermaphrodite ascaroside profiles are dominated by ascr#3, containing an α,β-unsaturated fatty acid, males predominantly produce the corresponding dihydro-derivative ascr#10. This small structural modification profoundly affects signaling properties: hermaphrodites are retained by attomole-amounts of male-produced ascr#10, whereas hermaphrodite-produced ascr#3 repels hermaphrodites and attracts males. Male production of ascr#10 is population density-dependent, indicating sensory regulation of ascaroside biosynthesis. Analysis of gene expression data supports a model in which sex-specific regulation of peroxisomal β-oxidation produces functionally different ascaroside profiles.


How to Build a Dichoptic Presentation System That Includes an Eye Tracker.
The presentation of different stimuli to the two eyes, dichoptic presentation, is essential for studies involving 3D vision and interocular suppression. There is a growing literature on the unique experimental value of pupillary and oculomotor measures, especially for research on interocular suppression. Although obtaining eye-tracking measures would thus benefit studies that use dichoptic presentation, the hardware essential for dichoptic presentation (e.g. mirrors) often interferes with high-quality eye tracking, especially when using a video-based eye tracker. We recently described an experimental setup that combines a standard dichoptic presentation system with an infrared eye tracker by using infrared-transparent mirrors1. The setup is compatible with standard monitors and eye trackers, easy to implement, and affordable (on the order of US$1,000). Relative to existing methods it has the benefits of not requiring special equipment and posing few limits on the nature and quality of the visual stimulus. Here we provide a visual guide to the construction and use of our setup.


Computed tomography of synovial sarcoma of the foot.
We report two patients with synovial sarcoma of the foot. Both presented with a soft-tissue mass that exhibited calcification. Although the radiologic findings are non-specific, the presence of foci of calcification within the soft-tissue mass is suggestive of synovial sarcoma. Computed tomography shows the extent of the mass, soft-tissue calcification and bone destruction thus helping in earlier detection and more accurate staging.


Continuous Monitoring of Pulse Oximetry During the 6-Minute Walk Test Improves Clinical Outcomes Prediction in COPD.
Continuous monitoring of SpO2 throughout the 6-min walk test (6MWT) unveiled that some patients with respiratory diseases may present values across the test lower than SpO2 measured at the end of the test. Nevertheless, it remains unclear whether this approach improves the yield of walk-induced desaturation detection in predicting mortality and hospitalizations in patients with COPD. Four hundred twenty-one subjects (51% males) with mild-very severe COPD underwent a 6MWT with continuous measurement of SpO2 . Exercise desaturation was defined as a fall in SpO2 ≥ 4%. All-cause mortality was assessed up to 6 y of follow-up and the rate of hospitalizations in the year succeeding the 6MWT. One hundred forty-nine subjects (35.4%) died during a mean (interquartile) follow-up of 55.5 (30.2-64.1) months. Desaturation was observed in 299/421 (71.0%). SpO2 along the test was < end SpO2 (88 [82-92]% vs 90 [84-93]%, P < .001). Desaturation detected only during (but not at the end of) the test was found in 81/421 (19.2%) participants. Multivariate Cox regression model adjusted for sex, body composition, FEV1, residual volume/total lung capacity ratio, walk distance, O2 supplementation during the test, and comorbidities retained the presence of desaturation either at the end (1.85 [95% CI 1.02-3.36]) or only along the test (2.08 [95% CI 1.09-4.01]) as significant predictors of mortality. The rate of hospitalizations was higher in those presenting with any kind of desaturation compared to those without exercise desaturation. Logistic regression analysis revealed that walking interruption and diffusing capacity of the lung for carbon monoxide predicted desaturation observed only during the test. O2 desaturation missed by end-exercise SpO2 but exposed by measurements during the test was independently associated with all-cause mortality and hospitalizations in subjects with COPD.


Emergence of human pandemic O25:H4-ST131 CTX-M-15 extended-spectrum-beta-lactamase-producing Escherichia coli among companion animals.
In view of the intercontinental emergence of Escherichia coli clone O25:H4-ST131 producing CTX-M-15 extended-spectrum beta-lactamase (ESBL) in human clinical settings it would be of great interest to explore its existence in animals to unravel a possible reservoir function and the origin and transmission of this group of multiresistant strains. A total of 177 clinical phenotypically ESBL-producing E. coli isolates, mainly obtained from companion animals with urinary tract infections, wound infections and diarrhoea, were collected in a veterinary diagnostic laboratory covering a European-wide service area. They were screened for molecular subtype O25b and multilocus sequence type 131. O25b-ST131 isolates were subsequently tested for ESBL types, and phenotypic and genotypic resistance determinants. Further characterization of the strains was performed by PFGE and virulence gene typing. Ten (5.6%) of 177 phenotypically ESBL-producing E. coli isolates, nine strains from dogs and one strain from a horse, were allocated to the B2-O25b-ST131 lineage. Nine of these isolates harboured a CTX-M-15-type beta-lactamase enzyme while one strain possessed an SHV-12-type ESBL. Macrorestriction analysis revealed a cluster formation of six of the animal CTX-M-15-type ESBL-producing strains from five different European countries together with a human control strain constituting a group of clonally related strains at a similarity value of 87.0%. Our findings demonstrate that the group of clonally related human B2-O25:H4-ST131 CTX-M-15-type ESBL-producing E. coli strains is present in companion animals from various European countries. This highlights the possibility of inter-species transmission of these multiresistant strains from human to animal and vice versa.