CryoEM structures of the human CLC-2 voltage gated chloride channel reveal a ball and chain gating mechanism.
CLC-2 is a voltage-gated chloride channel that contributes to electrical excitability and ion homeostasis in many different mammalian tissues and cell types. Among the nine mammalian CLC homologs, CLC-2 is uniquely activated by hyperpolarization, rather than depolarization, of the plasma membrane. The molecular basis for the divergence in polarity of voltage gating mechanisms among closely related CLC homologs has been a long-standing mystery, in part because few CLC channel structures are available, and those that exist exhibit high conformational similarity. Here, we report cryoEM structures of human CLC-2 at 2.46 - 2.76 Å, in the presence and absence of the potent and selective inhibitor AK-42. AK-42 binds within the extracellular entryway of the Cl - -permeation pathway, occupying a pocket previously proposed through computational docking studies. In the apo structure, we observed two distinct apo conformations of CLC-2 involving rotation of one of the cytoplasmic C-terminal domains (CTDs). In the absence of CTD rotation, an intracellular N-terminal 15-residue hairpin peptide nestles against the TM domain to physically occlude the Cl - -permeation pathway from the intracellular side. This peptide is highly conserved among species variants of CLC-2 but is not present in any other CLC homologs. Previous studies suggested that the N-terminal domain of CLC-2 influences channel properties via a "ball-and-chain" gating mechanism, but conflicting data cast doubt on such a mechanism, and thus the structure of the N-terminal domain and its interaction with the channel has been uncertain. Through electrophysiological studies of an N-terminal deletion mutant lacking the 15-residue hairpin peptide, we show that loss of this short sequence increases the magnitude and decreases the rectification of CLC-2 currents expressed in mammalian cells. Furthermore, we show that with repetitive hyperpolarization WT CLC-2 currents increase in resemblance to the hairpin-deleted CLC-2 currents. These functional results combined with our structural data support a model in which the N-terminal hairpin of CLC-2 stabilizes a closed state of the channel by blocking the cytoplasmic Cl - -permeation pathway.


HCMV Envelope Glycoprotein Diversity Demystified.
Human cytomegalovirus (HCMV) is the leading viral cause of congenital birth defects and is responsible for morbidity and mortality in immunosuppressed individuals. Considerable efforts have been deployed over the last decade to develop a vaccine capable of preventing HCMV infection. However, in recent clinical trials, vaccines showed at best modest efficacy in preventing infection. These findings might be explained by the high level of sequence polymorphism at the genomic level. To investigate if genomic variation also leads to antigenic variation, we performed a bioinformatic sequence analysis and evaluated the percentage of conservation at the amino acid level of all the proteins present in the virion envelope. Using more than two hundred sequences per envelope glycoprotein and analyzing their degree of conservation, we observe that antigenic variation is in large part limited to three proteins. In addition, we demonstrate that the two leading vaccine candidates, the pentamer and gB complexes, are well conserved at the amino acid level. These results suggest that despite genomic polymorphism, antigenic variability is not involved in the modest efficacy observed in the recent clinical trials for a HCMV vaccine. We therefore propose that next-generation vaccines should focus on stabilizing and refining the gB domains needed to induce a protective humoral response.


Neurons in ventrobasal region of cat thalamus selectively responsive to noxious mechanical stimulation.
1. A survey was made of neurons located in the ventral posterior lateral nucleus of the cat thalamus and its immediate vicinity for elements with specifically nociceptive properties. 2. Pipette microelectrodes filled with a dye solution were used to obtain extracellular recordings of unitary activity in 34 animals anesthetized with chloralose. 3. The great majority of the over 1,000 different single units responding to sciatic nerve stimulation noted in this series of experiments could also be excited by innocuous mechanical stimulation of skin or subcutaneous tissues. An infrequent but consistently noted group of units excited by A-alpha beta delta sciatic nerve volleys did not respond to innocuous mechanical manipulation or A-alpha beta sciatic nerve volleys; they were excited only by either noxious levels of mechanical stimulation or when volleys included the activity of more slowly conducting myelinated fibers. The latencies of such "high-threshold" units to sciatic volleys were longer than those of the other units. 4. Histologically identified recording sites marked by dye were recovered for 17 high-threshold units. Twelve of the 17 could be excited by noxious manipulations of restricted parts of the contralateral hindlimb. Nine of the 12 had cutaneous receptive fields, whereas 3 responded only to stimulation of subcutaneous tissues. None of the 17 high-threshold units evidenced additional discharges that could be correlated with the C-fiber component of sciatic nerve volleys. 5. The high-threshold units typically exhibited a low level of irregular background activity, which increased on repeated noxious stimulation of the peripheral receptive fields. Tactile units of the same or adjacent penetrations usually had a much greater degree of ongoing activity, often marked by bursts at a relatively high frequency. 6. The recording sites for the 17 high-threshold neurons were located dorsal and ventrolateral to the core of the ventrobasal nuclei and were not found in the midst of the low-threshold, cutaneous, mechanoreceptive population. During vertical stereotaxic penetrations, high-threshold units were noted dorsal or ventral to the location of ventrobasal tactile units in a pattern consistent with the core's somatotopic arrangement. 7. These results support the concept that the cat ventrolateral thalamus receives a small but distinct selectively nociceptive projection. The nociceptive neurons appear to be located in a shell that surrounds the main tactile projection to the ventral posterior lateral nucleus and that retains at least part of the topographic arrangement characteristic of the tactile core. Presumably, this projection is part of an organization identifying and localizing noxious stimulation.


Assessment of total phenolic and flavonoid content, antioxidant properties, and yield of aeroponically and conventionally grown leafy vegetables and fruit crops: a comparative study.
A comparison of the product yield, total phenolics, total flavonoids, and antioxidant properties was done in different leafy vegetables/herbs (basil, chard, parsley, and red kale) and fruit crops (bell pepper, cherry tomatoes, cucumber, and squash) grown in aeroponic growing systems (AG) and in the field (FG). An average increase of about 19%, 8%, 65%, 21%, 53%, 35%, 7%, and 50% in the yield was recorded for basil, chard, red kale, parsley, bell pepper, cherry tomatoes, cucumber, and squash, respectively, when grown in aeroponic systems, compared to that grown in the soil. Antioxidant properties of AG and FG crops were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DDPH) and cellular antioxidant (CAA) assays. In general, the study shows that the plants grown in the aeroponic system had a higher yield and comparable phenolics, flavonoids, and antioxidant properties as compared to those grown in the soil.


[Diaphorase reactions of lipoamide dehydrogenases from the adrenal ketoglutarate dehydrogenase complex].
Lipoamide dehydrogenase, a component of the bovine adrenal ketoglutarate dehydrogenase complex, catalyzes the oxidation of NADH by p-quinones and ferricyanide. The kinetics of oxidation obey the ping-pong mechanism. At pH 7.0, the constants for the active center oxidation by quinones (kox) are equal to 1.1 X 10(4)-5.3 X 10(5) M-1s-1 and increase as the acceptor potential rises. The values of kox for quinones change insignificantly within the pH range of 7.7-5.0, whereas that for ferricyanide increases 10-fold with a decrease of pH from 7.0 to 5.0. The value of the catalytic constant for the enzyme (kcat) reaches its maximum at pH 5.5. The quinones interact with the thiol groups of lipoamide dehydrogenase by inhibiting the fluorescence of FAD and diaphorease activity. The reaction is catalyzed by a basic amino acid (pK 6.7) within the composition of the enzyme.


Epigenetics of epithelial-to-mesenchymal transition in pancreatic carcinoma.
Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignant diseases with late diagnosis, rapid progression, high invasiveness, and early metastasis. Epithelial-to-mesenchymal transition (EMT) is a crucial step in metastasis that enables polarized immotile epithelial cells to gain fibroblast-like mesenchymal abilities such as enhanced motility. The dynamic process of EMT in PDAC with its powerful influence on disease progression and especially metastasis is of vigorous interest in biomedical research to elucidate its signaling pathways and regulation mechanisms. It is evident that epigenetics such as histone and DNA modification or noncoding RNAs such as microRNAs and long noncoding RNAs are of high importance in initiation and progress of EMT in PDAC. This review analyzes the latest research dealing with EMT and its epigenetic regulation in PDAC and summarizes its potentials in diagnostic, prognostic, and therapeutic management.


Lung dose calculations at kilovoltage x-ray energies using a model-based treatment planning system.
The determination of the dose to organs from diagnostic x rays has become important because of reports of radiation injury to patients from fluoroscopically guided interventional procedures. We have modified a convolution/superposition-based treatment planning system to compute the dose distribution for kilovoltage beams. We computed lung doses using this system and compared them to those calculated using the CDI3 organ dose calculation program. We also computed average lung doses from a simulated radiofrequency ablation procedure and compared our results to published doses for a similar procedure. Doses calculated using this system were an average of 20% lower for AP beams and 7% higher for PA beams than those obtained using CDI3. The ratio of the average dose to the lungs to the skin dose from the simulated ablation procedure ranged from 25% higher to 15% lower than that determined by other authors. Our results show that a treatment planning system designed for use in the megavoltage energy range can be used for calculating organ doses in the diagnostic energy range. Our doses compare well with those previously reported. Differences are partly due to variations in experimental techniques. Using a three-dimensional (3-D) treatment planning system to calculate dose also allows us to generate dose volume histograms (DVH) and compute normal tissue complication probabilities (NTCP) for diagnostic procedures.


Systematic Review, Meta-analysis, and Network Meta-analysis of the Cardiovascular Safety of Macrolides.
Studies reporting an increased risk for cardiac toxicities with macrolide antibiotics have raised concern regarding their cardiovascular safety. We sought to assess the cardiac safety of macrolide antibiotics as a class and of the individual agents by conducting a systematic review and network meta-analysis. Medline, Embase, and the Cochrane Library were searched up to February 2018 for studies reporting on cardiovascular outcomes with macrolides. We followed the PRISMA 2009 guidelines for data selection and extraction. Outcomes were pooled using random-effects models and odds ratios (OR), and 95% confidence intervals (CI) were calculated for arrhythmia, cardiovascular death, and myocardial infarction (MI). A total of 33 studies and data on 22,601,032 subjects were retrieved and included in the current meta-analyses. Macrolide use was not associated with the risk of arrhythmia or cardiovascular mortality. In the primary analysis, macrolide use was associated with a small but statistically significant 15% increase in risk for MI (OR = 1.15 [95% CI, 1.01 to 1.30]). In indirect network meta-analysis, erythromycin and clarithromycin were ranked considerably more likely to be associated with a higher risk for MI and significantly associated with increased risk of MI compared to azithromycin (OR = 1.58 [95% CI, 1.18 to 2.11] and OR = 1.41 [95% CI, 1.11 to 1.81], respectively). Our findings indicate that macrolide antibiotics as a group are associated with a significant risk for MI but not for arrhythmia and cardiovascular mortality. Among the macrolides, erythromycin and clarithromycin were associated with a greater risk of MI. However, it is possible that the association between macrolide use and risk of MI is the result of residual confounding.


Genetic hybridization of the leu-ilv region in bacilli.
Two auxotrophic strains of Bacillus subtilis 168 served as recipients for DNA extracted from various wild-type strains of B. subtilis and wild-type species of the genus Bacillus. Depending upon the DNA source, heterologous transformations of the linked try-his-tyr loci were either as efficient as those observed with donor DNA obtained from the wild type B. subtilis 168 strain or were undetectable. The order and relative distances of the three gene loci were the same for all active DNA preparations. Similar results were obtained in heterologous transformations of the linked leu-ilv loci, except that DNA preparations from the Bacillus globigii and B. subtilis var. niger species exhibited a reduced but detectable frequency of transformation. With the latter preparations a marked polarity of integration favouring the leu+ gene was observed, an effect not seen with homologous DNA. Six independent hybrid lines were obtained from transformation of B. subtilis leu- ilv- with DNA from B. subtilis var. niger leu+ ilv+. DNA extracted from these lines fell into two classes on the basis of activity in transforming the parental recipient strain: (i) indistinguishable from homologous DNA, and (ii) intermediate between homologous DNA and DNA from the original donor strain. With either class, polarity of integration was no longer observed in the leu-ilv region. The intermediate type of hybrid demonstrates that at least some of the inefficiency of heterospecific transformation must be due to heterology in nucleotide sequence between the different species at the leu-ilv loci.


Telomere length abnormalities in mammalian radiosensitive cells.
Telomere lengths in radiosensitive murine lymphoma cells L5178Y-S and parental radioresistant L5178Y cells were measured by quantitative fluorescence in situ hybridization. Results revealed a 7-fold reduction in telomere length in radiosensitive cells (7 kb) in comparison with radioresistant cells (48 kb). Therefore, it was reasoned that telomere length might be used as a marker for chromosomal radiosensitivity. In agreement with this hypothesis, a significant inverse correlation between telomere length and chromosomal radiosensitivity was observed in lymphocytes from 24 breast cancer patients and 5 normal individuals. In contrast, no chromosomal radiosensitivity was observed in mouse cell lines that showed shortened telomeres, possibly reflecting differences in radiation responses between primary cells and established cell lines. Telomere length abnormalities observed in radiosensitive cells suggest that these two phenotypes may be linked.


Lack of association between common endothelial nitric oxide synthase gene haplotypes and left ventricular hypertrophy in hypertension.
The endothelial nitric oxide synthase (NOS3) gene has been implicated in the pathogenesis of hypertension-related left ventricular hypertrophy (LVH). Candidate-gene studies have examined the role of NOS3 variation, but reported results are inconsistent. In this study, we investigated the association of three clinically relevant polymorphisms (promoter T786C, intronic 4a/b, and nonsynonymous G894T) in a case-control sample of 230 ethnically homogeneous (Caucasians) patients with essential hypertension, with (n = 64) and without (n = 166) clinically diagnosed LVH. Haplotype analysis was also performed. In single-marker analyses, no significant associations with LVH were detected by univariate and multivariate regression models. In the haplotype-based association analysis, no common haplotype was associated with the development of LVH. A rare haplotype consisting of the three mutant alleles (C-a-T*) was found to be present only in patients with LVH (3.4%) and not in control hypertensive patients. Despite the biological rationale for the involvement of the NOS3 gene in LVH, no evidence for a major role of common NOS3 haplotypic variation was found. Considering the totality of available evidence, single-gene analyses of the NOS3 gene have not uncovered detectable genetic effects, and pathway-based analyses that examine interactions of multiple loci may be more informative about the complex genetic etiology of LVH.


X-irradiation of Eimeria tenella oocysts provides direct evidence that sporozoite invasion and early schizont development induce a protective immune response(s).
Sporulated oocysts of the protozoan parasite Eimeria tenella were attenuated by exposure to various doses of X-radiation to inhibit intracellular replication and thus determine whether sporozoites alone can induce a protective immune response. Exposure to doses greater than 15-kilorads had a significant effect on development, as indicated by the absence of oocyst production in chickens infected with parasites treated with 20 or 30 kilorads of radiation. Infection with nonirradiated or 15-kilorad-exposed parasites led to either normal or reduced oocyst shedding. Equivalent protection was afforded chickens inoculated with a minimum immunizing dose of either nonirradiated or 20-kilorad-irradiated E. tenella oocysts. Immunofluorescence staining of cecal tissue from chickens inoculated with 10(7) nonirradiated or 20- or 30-kilorad-irradiated oocysts with stage-specific monoclonal antibodies showed no significant difference in sporozoite invasion between treatment groups. Normal merogonic development was observed at appropriate times (48, 60, 72, and 96 h) postinfection in chickens inoculated with nonirradiated oocysts. In contrast, irradiated parasites exhibited minimal merogonic development at 48 h postinfection. Furthermore, no merogonic stages were observed at times of otherwise peak merozoite development (60, 72, and 96 h) in cecal tissue from chickens inoculated with irradiated parasites. Infection of chicken cells with irradiated or nonirradiated parasites in vitro corroborated these findings and indicate that events early after sporozoite invasion induce a protective immune response against this parasite.


Characterization of 6-phosphofructo-2-kinase from foetal-rat liver.
Foetal and adult liver 6-phosphofructo-2-kinase (PFK-2) were purified by identical protocols. The native molecular masses of both enzymes were determined by gel filtration and were 89.1 and 100.0 kDa respectively. No differences were found in SDS/PAGE in 10%-acrylamide gel (55 kDa per subunit). The kinetic properties displayed by both enzymes were similar, except for the sensitivity to inhibition by sn-glycerol 3-phosphate. Foetal PFK-2 was a good substrate for phosphorylation by cyclic AMP-dependent protein kinase and protein kinase C, whereas the adult enzyme was phosphorylated only by cyclic AMP-dependent protein kinase. However, the phosphorylation affected only the kinetic properties of the adult enzyme, suggesting the presence in both enzymes of different sites of phosphorylation by cyclic AMP-dependent protein kinase. These differences in primary structure were consistent with the distinct chromatographic profiles of the phosphopeptides after digestion of the protein with CNBr. Western-blot analysis with antibodies specific for the N-terminal region of the liver-type PFK-2 poorly recognized the foetal enzyme, suggesting that both enzymes differ at least in the N-terminal sequence.


A two-cycle prospective audit of temporal bone computed tomography scan requests: improving the clinical applicability of radiology reports.
Radiologists require accurate clinical information to formulate reports. This is particularly relevant to computed tomography of the temporal bone, in which previous surgery can mimic disease. The information provided with temporal bone computed tomography scan requests was evaluated. The study aimed to minimise inappropriate requests and improve the clinical value of reports. A two-cycle prospective audit was undertaken using a proforma designed on the basis of national guidelines. Following the first cycle (in which the requests and reports of 100 scans were evaluated), new guidelines and training were implemented. A follow-up audit (of 50 scans) was then performed. Following intervention, the percentage of clinically relevant reports increased from 52 to 94 (p < 0.01), whilst unnecessary or inappropriate scan requests decreased from 11 to 2 per cent (p < 0.05). Optimising the clinical value of temporal bone computed tomography scan requests will have positive implications for patient care, time management and cost. The quality of the clinical information provided can have a significant impact on the clinical value of radiology reports, and can mean that unnecessary irradiation is avoided.


Distribution of ADH2 and ALDH2 genotypes in different populations.
The distribution of the human liver alcohol dehydrogenase, ADH2, and aldehyde dehydrogenase, ALDH2, genotypes in 21 different populations comprising Mongoloids, Caucasoids, and Negroids was determined by hybridization of the amplified genomic DNA with allele-specific oligonucleotide probes. Whereas the frequency of the ADH1(2) allele was found to be relatively high in the Caucasoids, Mexican Mestizos, Brazilian Indios, Swedish Lapps, Papua New Guineans and Negroids, the frequency of the ADH2(2) gene was considerably higher in the Mongoloids and Australian Aborigines. The atypical ALDH2 gene (ALDH2(2)) was found to be extremely rare in Caucasoids, Negroids, Papua New Guineans, Australian Aborigines and Aurocanians (South Chile). In contrast, this mutant gene was found to be widely prevalent among the Mongoloids. Individuals possessing the abnormal ALDH2 gene show alcohol-related sensitivity responses (e.g. facial flushing), have the tendency not to be habitual drinkers, and apparently suffer less from alcoholism and alcohol-related liver disease.


Copper-doped mesoporous hydroxyapatite microspheres synthesized by a microwave-hydrothermal method using creatine phosphate as an organic phosphorus source: application in drug delivery and enhanced bone regeneration.
The development of multifunctional biomaterials with drug delivery ability, and pro-osteogenic and pro-angiogenic activities has garnered increasing interest in the field of regenerative medicine. In the present study, hypoxia-mimicking copper (Cu)-doped mesoporous hydroxyapatite (HAP) microspheres (Cu-MHMs) were successfully synthesized through a microwave-hydrothermal method by using creatine phosphate as an organic phosphorus source. The Cu-MHMs doped with 0.2, 0.5 and 1 mol% Cu were prepared. The Cu-MHMs consisting of HAP nanorods or nanosheets exhibited a hierarchically mesoporous hollow structure and a high specific surface area. Then the Cu-MHMs were investigated as a drug nanocarrier using doxorubicin hydrochloride (DOX) as a model drug. The Cu-MHMs showed a relatively high drug-loading capacity and a pH-responsive drug release behavior. Furthermore, the Cu-MHMs were incorporated into a chitosan (CS) matrix to construct a biomimetic scaffold optimized for bone regeneration. The Cu-MHM/CS composite scaffolds maintained high degrees of porosity and showed a sustained release of Cu ions. More importantly, the Cu-MHM/CS scaffolds not only enhanced the osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (rBMSCs) but also promoted the migration and tube formation of EA.hy926 cells. When implanted in rat critical-sized calvarial defects, the Cu-MHM/CS scaffolds significantly enhanced bone regeneration accompanied by more new blood vessel formation at 8 weeks post-operation compared with the MHM/CS scaffolds. These results suggest that the hypoxia-mimicking Cu-MHM/CS scaffolds could encourage bone regeneration by enhancing osteogenesis and angiogenesis simultaneously, which bodes well for the reconstruction of vascularized tissue-engineered bone.


Bcl-2 mutants with restricted subcellular location reveal spatially distinct pathways for apoptosis in different cell types.
Human Bcl-2 is located in multiple intracellular membranes when expressed in MDCK and Rat-1/myc cells. We restricted expression to the endoplasmic reticulum or mitochondria by exchanging the Bcl-2 carboxy-terminal insertion sequence for an equivalent sequence from cytochrome b5 or ActA, respectively. MDCK cells are protected from serum deprivation-induced apoptosis by both wild-type Bcl-2 and the mutant targeted to mitochondria but not by the mutant targeted to endoplasmic reticulum. In contrast, when expressed in Rat-1/myc cells, the Bcl-2 mutant located at the endoplasmic reticulum is more effective than that targeted to mitochondria. In MDCK cells both mutants bind Bax as effectively as wild-type, demonstrating that Bax binding is not sufficient to prevent apoptosis.


Myelopathy and Quadriparesis due to Spinal Cord Compression of C1 Laminar Osteochondroma.
The aim of this paper is to show that osteochondromas of the cervical vertebrae can cause myelopathy and neck pain.The reported etiology, diagnosis, treatment and differential diagnosis were reviewed. Osteochondromas may present as a solitary lesion with no genetic component or as multiple lesions as a part of a genetic disorder known as hereditary multiple exostosis. Osteochondromas of the spine are rarely encountered in clinical practice. These lesions are reported more commonly with neural compression in cases associated with hereditary multiple exostosis. The authors describe a unusual clinical manifestation of a solitary osteochondroma located in the right posterior arch of the atlas. Complete removal of the tumor was performed resulting in the relief of neck pain and spastic quadriparesis. Although unusual, osteochondromas of the cervical spine must be considered in patients with persistent neck pain and progressive symptoms of myelopathy. Computed tomography and magnetic resonance imaging in conjunction with plain radiograms is the neuroradiological modality of choice. The diagnosis and surgical excision of these tumors are important because they can cause spinal stenosis resulting in neural tissue compression and myelopathy.


Mortality implications of lower DBP with lower achieved systolic pressures in coronary artery disease: long-term mortality results from the INternational VErapamil-trandolapril STudy US cohort.
A goal SBP 120 mmHg or less reduced mortality in high-risk Systolic Blood Pressure Intervention Trial patients; however, mortality implications of concomitant DBP lowering in coronary artery disease (CAD) are uncertain. We examined the relationship between DBP lowering and all-cause mortality with lower achieved SBPs in a large cohort. We categorized 17 131 hypertensive patients from the INternational VErapamil-trandolapril STudy US cohort, aged at least 50 years with CAD, by mean achieved SBP (<120, 120 to <130, 130 to <140, and ≥140 mmHg) and DBP tertiles (low, middle, and high per SBP category) during active follow-up. Long-term mortality was determined via National Death Index. Multivariable Cox regression was performed to investigate the impact of DBP lowering among all SBP categories and within each SBP category. There were 6031 deaths over mean follow-up of 11.6 years (198 352 patient-years). In unadjusted analyses, achieving DBP in the lowest tertile portended greatest mortality risk across all SBP categories. In multivariate analysis, using SBP 120 to less than 130 mmHg, DBP at least 79 mmHg as reference (mortality nadir), achieving DBP in the lowest tertile (DBP < 69 mmHg) was associated with excess mortality risk among those with SBP less than 120 mmHg (adjusted hazard ratio 1.60; 95% confidence interval, 1.33-1.91). However, among those with SBP 120 to less than 140 mmHg, adjusted mortality risk did not differ significantly with low DBPs. Among those with SBP at least 140 mmHg, mortality risk remained high regardless of DBP. In older CAD patients, the mortality risk related to excess DBP lowering is accentuated in those achieving intensive SBP control less than 120 mmHg, raising concerns about intensive SBP lowering in these patients.


Clinical and Laboratory Profile of COVID-19 Pneumonia Patients With a Complicated Post-Intensive Care Unit Hospital Course.
Characteristics of intensive care unit (ICU) downgrades who experience a complicated post-ICU ward course (ICU return or floor death) and the incidence of this phenomenon have not been examined in ICU survivors of coronavirus disease 2019 (COVID-19) pneumonia. The aim of the present study was to establish the rate of a complicated post-ICU ward course among survivors of COVID-19 pneumonia and describe the associated patient, ICU management, and serum biomarker characteristics. An additional aim was to compare these parameters between those who experienced a complicated post-ICU course and those who did not. This was a retrospective study of patients who were admitted to the ICU with COVID-19 pneumonia and were downgraded to a hospital floor at the end of their initial ICU stay. Patients were divided based on a complicated or uncomplicated post-ICU course. Groups were compared with respect to relevant clinical variables. Serum biomarker levels were compared on day of ICU exit and were trended in the days preceding the downgrade. Ward stay of patients who had a complicated course was examined for notable floor events surrounding their decompensation. Eighteen out of 99 downgraded patients (18%) experienced a complicated post-ICU course, among them there were 14 returns (14%) and four deaths (4%). They had higher Charlson Comorbidity Index, higher Acute Physiology and Chronic Health Evaluation (APACHE) IV score, as well as higher D-dimer and C-reactive protein (CRP) at ICU departure. They were less likely to have received therapeutic anticoagulation and convalescent plasma during their ICU stay. On multivariable analysis, these parameters except D-dimer remained independently associated with a complicated course. Review of biomarker trends preceding ICU exit demonstrated an upward trajectory of D-dimer, CRP, and lactate dehydrogenase (LDH) in the complicated course group not mirrored by the uncomplicated course group. Examination of notable floor events leading up to decompensation revealed that in 50% the ward course was characterized by new cardiac disturbances. Our rate of ward death among ICU downgrades was similar to pre-COVID data, but the rate of ICU return was higher. Complicated post-ICU course patients were exhibiting upward biomarker trends at ICU exit, and their ward stay was punctuated by acute cardiac abnormalities.


Classifying dengue: a review of the difficulties in using the WHO case classification for dengue haemorrhagic fever.
The current World Health Organisation (WHO) classification of dengue includes two distinct entities: dengue fever (DF) and dengue haemorrhagic fever (DHF)/dengue shock syndrome; it is largely based on pediatric cases in Southeast Asia. Dengue has extended to different tropical areas and older age groups. Variations from the original description of dengue manifestations are being reported. To analyse the experience of clinicians in using the dengue case classification and identify challenges in applying the criteria in routine clinical practice. Systematic literature review of post-1975 English-language publications on dengue classification. Thirty-seven papers were reviewed. Several studies had strictly applied all four WHO criteria in DHF cases; however, most clinicians reported difficulties in meeting all four criteria and used a modified classification. The positive tourniquet test representing the minimum requirement of a haemorrhagic manifestation did not distinguish between DHF and DF. In cases of DHF thrombocytopenia was observed in 8.6-96%, plasma leakage in 6-95% and haemorrhagic manifestations in 22-93%. The low sensitivity of classifying DHF could be due to failure to repeat the tests or physical examinations at the appropriate time, early intravenous fluid therapy, and lack of adequate resources in an epidemic situation and perhaps a considerable overlap of clinical manifestations in the different dengue entities. A prospective multi-centre study across dengue endemic regions, age groups and the health care system is required which describes the clinical presentation of dengue including simple laboratory parameters in order to review and if necessary modify the current dengue classification.


The Footfall Programme: participant experiences of a lower limb, foot and ankle exercise intervention for falls prevention - an exploratory study.
Despite growing evidence that foot and ankle exercise programmes are effective for falls prevention, little is known about older adults' views and preferences of programme components for long-term maintenance. The aims of this study were to explore the experiences and acceptability of Scottish and Portuguese older adults of undertaking a home-based foot, ankle and lower limb exercise intervention. Ten Scottish (mean age 76 years, 7 female) and fourteen Portuguese (mean age 66 years, 12 female) community-dwelling older adults undertook the programme for one week, followed by focus group discussions (2-6 people per group), guided by a semi-structured interview guide. Data was analysed using thematic analysis. Seven themes were identified:Assessment, Group exercise taster, Home based exercise; Footfall programme kit, Midweek phone call, Reasons for participation and the Research Process. Programme components, support telephone calls and research procedures were generally well accepted by participants and they valued having a contribution to the design. They preferred a blended home and intermittent group-based programme format for motivation and progression and recommended changes to some of the exercises and equipment to reduce barriers to participation. Some cultural differences emerged, including importance of the functional assessments for Portuguese participants, time issues and difficulty in completion of the exercise diary, reflecting lower literacy levels. Participants found the programme acceptable but preferred a blended home and occasional group-based programme for adherence and motivation. A strong educational component to improve health literacy and simple paperwork completion to avoid data loss in future studies with Portuguese older adults is important.


Subcortical heterotopia: a distinct clinicoradiologic entity.
To investigate the clinical syndrome and associated brain anomalies in a group of patients with subcortical heterotopia. The seizure history, developmental history, family history, electroencephalographic results, imaging studies, and pathologic results (where available) of 13 patients with subcortical heterotopia were reviewed retrospectively. The patients ranged in age from 1 month to 39 years (mean, 11 years; median, 3 years) at the time of the most recent update of their records. Fifteen hemispheres were involved in 13 patients. The cerebral cortex overlying the heterotopia was thin with shallow sulci and the affected part of the hemisphere was small as compared with the contralateral hemisphere. The corpus callosum was hypogenetic or agenetic in nine patients and the basal nuclei were dysplastic or hypoplastic in 11 patients. Most patients were developmentally delayed as children, had mild hemiplegia or hemihypesthesia contralateral to the affected hemisphere(s), and had partial epilepsy, most commonly evidenced by partial motor seizures in the body half contralateral to the malformation. Age at seizure onset varied from infancy to the second decade. Electroencephalographic results showed slow wave background with spike and spike wave complexes in the affected hemisphere. Subcortical heterotopia seems to have characteristic clinical, electrophysiological, and imaging manifestations. These results suggest that affected patients may be conveniently grouped together to study treatment outcomes.


Emerging Role of AMPK in Brown and Beige Adipose Tissue (BAT): Implications for Obesity, Insulin Resistance, and Type 2 Diabetes.
The global prevalence of type 2 diabetes (T2D) is escalating at alarming rates, demanding the development of additional classes of therapeutics to further reduce the burden of disease. Recent studies have indicated that increasing the metabolic activity of brown and beige adipose tissue may represent a novel means to reduce circulating glucose and lipids in people with T2D. The AMP-activated protein kinase (AMPK) is a cellular energy sensor that has recently been demonstrated to be important in potentially regulating the metabolic activity of brown and beige adipose tissue. The goal of this review is to summarize recent work describing the role of AMPK in brown and beige adipose tissue, focusing on its role in adipogenesis and non-shivering thermogenesis. Ablation of AMPK in mouse adipocytes results in cold intolerance, a reduction in non-shivering thermogenesis in brown adipose tissue (BAT), and the development of non-alcoholic fatty liver disease (NAFLD) and insulin resistance; effects associated with a defect in mitochondrial specific autophagy (mitophagy) within BAT. The effects of a β3-adrenergic agonist on the induction of BAT thermogenesis and the browning of white adipose tissue (WAT) are also blunted in mice lacking adipose tissue AMPK. A specific AMPK activator, A-769662, also results in the activation of BAT and the browning of WAT, effects which may involve demethylation of the PR domain containing 16 (Prdm16) promoter region, which is important for BAT development. AMPK plays an important role in the development and maintenance of brown and beige adipose tissue. Adipose tissue AMPK is reduced in people with insulin resistance, consistent with findings that mice lacking adipocyte AMPK develop greater NAFLD and insulin resistance. These data suggest that pharmacologically targeting adipose tissue AMPK may represent a promising strategy to enhance energy expenditure and reduce circulating glucose and lipids, which may be effective for the treatment of NAFLD and T2D.


Integrating ecosystem markets to co-ordinate landscape-scale public benefits from nature.
Ecosystem markets are proliferating around the world in response to increasing demand for climate change mitigation and provision of other public goods. However, this may lead to perverse outcomes, for example where public funding crowds out private investment or different schemes create trade-offs between the ecosystem services they each target. The integration of ecosystem markets could address some of these issues but to date there have been few attempts to do this, and there is limited understanding of either the opportunities or barriers to such integration. This paper reports on a comparative analysis of eleven ecosystem markets in operation or close to market in Europe, based on qualitative analysis of 25 interviews, scheme documentation and two focus groups. Our results indicate three distinct types of markets operating from the regional to national scale, with different modes of operation, funding and outcomes: regional ecosystem markets, national carbon markets and green finance. The typology provides new insights into the operation of ecosystem markets in practice, which may challenge traditionally held notions of Payment for Ecosystem Services. Regional ecosystem markets, in particular, represent a departure from traditional models, by using a risk-based funding model and aggregating both supply and demand to overcome issues of free-riding, ecosystem service trade-offs and land manager engagement. Central to all types of market were trusted intermediaries, brokers and platforms to aggregate supply and demand, build trust and lower transaction costs. The paper outlines six options for blending public and private funding for the provision of ecosystem services and proposes a framework for integrating national carbon markets and green finance with regional ecosystem markets. Such integration may significantly increase funding for regenerative agriculture and conservation across multiple habitats and services, whilst addressing issues of additionality and ecosystem service trade-offs between multiple schemes.


Anti-NMDA receptor encephalitis: an easily missed diagnosis in older patients.
Anti-N-methyl-D-aspartate (NMDA) receptor encephalitis is an important, treatable cause of encephalitis which remains under-recognised despite a growing body of the literature [1]. It is an immune-mediated syndrome which presents with a variety of neurological symptoms including headache, fever, personality change and seizures. Most case reports to date are of young adults, it is much less frequently reported in older adults. The syndrome has been associated with ovarian teratomas. The prognosis is good with early recognition and treatment, though may relapse. We present a case of NMDA encephalitis in an elderly patient who responded well to immunosuppressive therapy.


Two-year longitudinal analysis of a cluster randomized trial of physical activity promotion by general practitioners.
We evaluate the effectiveness of a physical activity promotion programme carried out by general practitioners with inactive patients in routine care. Pragmatic, cluster randomised clinical trial conducted in eleven public primary care centres in Spain. Fifty-six general practitioners (GPs) were randomly assigned to intervention (29) or standard care (27) groups. They assessed the physical activity level of a systematic sample of patients in routine practice and recruited 4317 individuals (2248 intervention and 2069 control) who did not meet minimum physical activity recommendations. Intervention GPs provided advice to all patients and a physical activity prescription to the subgroup attending an additional appointment (30%). A third of these prescriptions were opportunistically repeated. Control GPs provided standard care. Primary outcome measure was the change in self-reported physical activity from baseline to six, 12 and 24 months. Secondary outcomes included cardiorespiratory fitness and health-related quality of life. A total of 3691 patients (85%) were included in the longitudinal analysis and overall trends over the whole 24 month follow-up were significantly better in the intervention group (p<0.01). The greatest differences with the control group were observed at six months (adjusted difference 1.7 MET*hr/wk [95% CI, 0.8 to 2.6], 25 min/wk [95% CI, 11.3 to 38.4], and a 5.3% higher percentage of patients meeting minimum recommendations [95% CI: 2.1% to 8.8%] NNT = 19). These differences were not statistically significant at 12 and 24 months. No differences were found in secondary outcomes. A significant difference was maintained until 24 months in the proportion of patients achieving minimum recommendation in the subgroup that received a repeat prescription (adjusted difference 10.2%, 95% CI 1.5% to 19.4%). General practitioners are effective at increasing the level of physical activity among their inactive patients during the initial six-months of an intervention but this effect wears off at 12 and 24 months. Only in the subgroup of patients receiving repeat prescriptions of physical activity is the effect maintained in long-term. clinicaltrials.gov NCT00131079.


High-Density Lipoprotein Functionality as a New Pharmacological Target on Cardiovascular Disease: Unifying Mechanism That Explains High-Density Lipoprotein Protection Toward the Progression of Atherosclerosis.
The formation of the atherosclerotic plaque that is characterized by the accumulation of abnormal amounts of cholesterol-loaded macrophages in the artery wall is mediated by both inflammatory events and alterations of lipid/lipoprotein metabolism. Reverse transport of cholesterol opposes the formation and development of atherosclerotic plaque by promoting high density lipoprotein (HDL)-mediated removal of cholesterol from peripheral macrophages and its delivery back to the liver for excretion into the bile. Although an inverse association between HDL plasma levels and the risk of cardiovascular disease (CVD) has been demonstrated over the years, several studies have recently shown that the antiatherogenic functions of HDL seem to be mediated by their functionality, not always associated with their plasma concentrations. Therefore, assessment of HDL function, evaluated as the capacity to promote cell cholesterol efflux, may offer a better prediction of CVD than HDL levels alone. In agreement with this idea, it has recently been shown that the assessment of serum cholesterol efflux capacity (CEC), as a metric of HDL functionality, may represent a predictor of atherosclerosis extent in humans. The purpose of this narrative review is to summarize the current evidence concerning the role of cholesterol efflux capacity that is important for evaluating CVD risk, focusing on pharmacological evidences and its relationship with inflammation. We conclude that HDL therapeutics are a promising area of investigation but strategies for identifying efficacy must move beyond the idea of simply raising static HDL-cholesterol levels and toward methods of measuring the dynamics of HDL particle remodeling and the generation of lipid-free apolipoprotein A-I (apoA-I). In this way, apoA-I, unlike mature HDL, can promote the greatest extent of cholesterol efflux relieving cellular cholesterol toxicity and the inflammation it causes.


Adult neurogenesis in the short-lived teleost Nothobranchius furzeri: localization of neurogenic niches, molecular characterization and effects of aging.
We studied adult neurogenesis in the short-lived annual fish Nothobranchius furzeri and quantified the effects of aging on the mitotic activity of the neuronal progenitors and the expression of glial fibrillary acid protein (GFAP) in the radial glia. The distribution of neurogenic niches is substantially similar to that of zebrafish and adult stem cells generate neurons, which persist in the adult brain. As opposed to zebrafish, however, the N. furzeri genome contains a doublecortin (DCX) gene. Doublecortin is transiently expressed by newly generated neurons in the telencephalon and optic tectum (OT). We also analyzed the expression of the microRNA miR-9 and miR-124 and found that they have complementary expression domains: miR-9 is expressed in the neurogenic niches of the telencephalon and the radial glia of the OT, while miR-124 is expressed in differentiated neurons. The main finding of this paper is the demonstration of an age-dependent decay in adult neurogenesis. Using unbiased stereological estimates of cell numbers, we detected an almost fivefold decrease in the number of mitotically active cells in the OT between young and old age. This reduced mitotic activity is paralleled by a reduction in DCX labeling. Finally, we detected a dramatic up-regulation of GFAP in the radial glia of the aged brain. This up-regulation is not paralleled by a similar up-regulation of S100B and Musashi-1, two other markers of the radial glia. In summary, the brain of N. furzeri replicates two typical hallmarks of mammalian aging: gliosis and reduced adult neurogenesis.


How older people with incurable cancer experience daily living: A qualitative study from Norway.
An increasing number of older people are living with incurable cancer as a chronic disease, requiring palliative care from specialized healthcare for shorter or longer periods of time. The aim of our study was to describe how they experience daily living while receiving palliative care in specialized healthcare contexts. We conducted a qualitative research study with a phenomenological approach called "systematic text condensation." A total of 21 participants, 12 men and 9 women, aged 70-88, took part in semistructured interviews. They were recruited from two somatic hospitals in southeastern Norway. The participants experienced a strong link to life in terms of four subthemes: to acknowledge the need for close relationships; to maintain activities of normal daily life; to provide space for existential meaning-making and to name and handle decline and loss. In addition, they reported that specialized healthcare contexts strengthened the link to life by prioritizing and providing person-centered palliative care. Older people with incurable cancer are still strongly connected to life in their daily living. The knowledge that the potential for resilience remains despite aging and serious decline in health is considered a source of comfort for older people living with this disease. Insights into the processes of existential meaning-making and resilience are seen as useful in order to increase our understanding of how older people adapt to adversity, and how their responses may help to protect them from some of the difficulties inherent to aging. Healthcare professionals can make use of this information in treatment planning and for identification of psychosocial and sociocultural resources to support older people and to strengthen patients' life resources.


Distal molar movement using the pendulum appliance. Part 1: Clinical and radiological evaluation.
Intra-arch distal molar movement techniques have recently assumed an important role in clinical orthodontics. In this study, the dental and skeletal effects of the pendulum appliance, applying 200 to 250 g of force to the molars in 13 patients (age range 8 years to 13 years 5 months) were evaluated by means of cephalometric radiographs. The results showed that the pendulum appliance moved the molars distally without creating dental or skeletal bite opening and with little incisor anchorage loss. However, important molar tipping should be taken into consideration when using this appliance.


Liquid chromatography-tandem mass spectrometry metabolic profiling of nazartinib reveals the formation of unexpected reactive metabolites.
Nazartinib (EGF816, NZB) is a promising third-generation human epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor. This novel irreversible mutant-selective EGFR inhibitor targets EGFR containing both the resistance mutation (T790M) and the activating mutations (L858R and Del19), while it does not affect wild-type EGFR. However, the metabolic pathway and bioactivation mechanisms of NZB are still unexplored. Thus, using liquid chromatography-tandem mass spectrometry, we screened for products of NZB metabolism formed in vitro by human liver microsomal preparations and investigated the formation of reactive intermediates using potassium cyanide as a nucleophile trap. Unexpectedly, the azepane ring was not bioactivated. Instead, the carbon atom between the aliphatic linear tertiary amine and electron-withdrawing system (butenoyl amide group) was bioactivated, generating iminium intermediates as reactive species. Six NZB phase I metabolites, formed by hydroxylation, oxidation and N-demethylation, were characterized. Moreover, two reactive iminium ions were characterized and their corresponding bioactivation mechanisms were proposed. Based on our results, we speculate that bioactivation of NZB can be blocked by small sterically hindering groups, isosteric replacement or a spacer. This approach might reduce the toxicity of NZB by avoiding the generation of reactive species.


Continuous-flow enzymatic determination of total serum cholesterol and method standardization with CDC-calibrated pooled sera.
We compared Boehringer Mannheim's enzymatic kit for the continuous-flow (AutoAnalyzer II) determination of serum cholesterol with Technicon's N-24a extraction method. Results for patients' samples analyzed by the enzymatic method were higher than those by the comparison method. To evaluate accuracy in the cholesterol determinations, we enrolled the enzymatic method into the Center for Disease Control's (CDC) Lipid Standardization program. We calibrated the method by use of a pooled sera for which cholesterol content was assigned by CDC after analysis by their reference Abell-Kendall procedure. We discuss the difficulties with available calibration material and limitations in the application of some commercial control materials to the enzymatic cholesterol method. The continuous-flow variables, Michaelis-Menton constants, percent ester-hydrolase activity, and other factors affecting the performance of the enzyme-linked cholesterol method are evaluated. We believe pooled sera with an assigned value for cholesterol content is the best calibrator material.


A Link Between Nerve Growth Factor Metabolic Deregulation and Amyloid-β-Driven Inflammation in Down Syndrome.
In Alzheimer's disease and Down syndrome, cholinergic neurons of the basal forebrain progressively degenerate. This neurotransmitter system is the main source of acetylcholine to the cortex and hippocampus. In the mature and fully differentiated central nervous system, the phenotype of forebrain cholinergic neurons and their nerve terminals in cortex and hippocampus depend on the continuous endogenous supply of nerve growth factor (NGF). It has been recently demonstrated that NGF is secreted from cortical neurons in an activity-dependent manner as a precursor molecule, proNGF. Individuals with Alzheimer's disease and Down syndrome exhibit proNGF accumulation in cortex, yet cholinergic neurons become atrophic in both diseases, despite the apparent abundance of the NGF precursor. This review illustrates the recent evidence that NGF metabolism is affected both in Alzheimer's disease and in Down syndrome brains and also discusses a role for amyloid-β peptides and central nervous system inflammation in unleashing such deficits. It further considers the potential of the NGF metabolic pathway as a new pharmacological target to slow down the neurodegenerative process both in Alzheimer's disease and in individuals with Down syndrome.


Clinical outcomes of patients with pT0 bladder cancer after radical cystectomy: a single-institute experience.
To investigate the clinical outcomes of patients who underwent radical cystectomy for bladder cancer at a single institution and compare those who had pT0 specimens with those who had residual cancer. From January 1990 to December 2006, 186 patients underwent radical cystectomy with or without neoadjuvant chemotherapy for cT2 or higher stage urothelial carcinoma in the bladder in our hospital. We estimated the 5-year disease-free survival, cancer-specific survival and overall survival by the pathological stage. The median follow-up of the 186 patients was 38.5 months (0-194). Of these, 51 received neoadjuvant chemotherapy. For all subjects, the 5-year disease-free survival was 54.9%, cancer-specific survival 61.0% and overall survival 57.1%. Of the 186 patients, 24 (12.9%) had no residual cancer in the bladder specimen at radical cystectomy. Of the 24 patients with pT0, only 1 (4.2%) died of bladder cancer. The 5-year disease-free survival, cancer-specific survival and overall survival rates in patients with pT0 were ∼96.0%. We found pT0 histology in 11 of the 51 patients (21.6%) with neoadjuvant chemotherapy and in 13 of the 135 patients (9.6%) with radical cystectomy alone (P = 0.047). We demonstrated that the outcomes of patients who underwent radical cystectomy were similar to those in previous reports. Patients with pT0 showed favorable outcomes for disease-free survival, cancer-specific survival and overall survival in our study. However, they should be periodically followed up because pT0 does not always mean cure.


Motoneuron electrophysiological and muscle contractile properties of superior oblique motor units in cat.
Intracellular techniques were used to study single motor units of the trochlear nucleus and superior oblique muscle in the cat. Motoneuron electrophysiological properties were correlated with muscle-unit contractile characteristics assessed under isometric conditions. Two distinct motor-unit types were identified and designated as twitch and nontwitch. Nontwitch units made up 5% of the total population studied. They responded only to tetanic stimulation with graded force that increased as stimulus frequency was increased up to 300-400 Hz. These units made up a homogeneous population in that they were innervated by slowly conducting axons, produced weak tetanic tensions, and were extremely fatigue resistant. Twitch units made up the majority (95%) of units studied. These units responded to single pulse stimulation with typical twitch contractions. The contraction speed and tension ranges for these units were comparable with those obtained from other extraocular muscle single units. Superior oblique twitch units, mechanically comparable with multiply innervated conducting units, identified in the cat inferior oblique muscle (31) were not observed. The twitch-unit population was heterogeneous in terms of neuromuscular fatigue resistance. Unit fatigability was inversely related to maximal tetanic tension. Motoneuron conduction velocity was related to muscle-unit contractile properties in a way similar to that seen in extremity motor units. The slowest twitch units were weak, fatigue resistant, and innervated by slow conducting axons. The fastest units were, in general, innervated by faster conducting axons, produced greater tetanic tensions, and were more susceptible to fatigue. Correlations among input resistance, rheobase, and conduction velocity were also observed. At present, subdivisions of the twitch-unit population on the basis of any one or combination of unit properties does not seem appropriate.


Cadherins and catenins in dendrite and synapse morphogenesis.
Neurons are highly polarized specialized cells. Neuronal integrity and functional roles are critically dependent on dendritic architecture and synaptic structure, function and plasticity. The cadherins are glycosylated transmembrane proteins that form cell adhesion complexes in various tissues. They are associated with a group of cytosolic proteins, the catenins. While the functional roles of the complex have been extensively investigates in non-neuronal cells, it is becoming increasingly clear that components of the complex have critical roles in regulating dendritic and synaptic architecture, function and plasticity in neurons. Consistent with these functional roles, aberrations in components of the complex have been implicated in a variety of neurodevelopmental disorders. In this review, we discuss the roles of the classical cadherins and catenins in various aspects of dendrite and synapse architecture and function and their relevance to human neurological disorders. Cadherins are glycosylated transmembrane proteins that were initially identified as Ca(2+)-dependent cell adhesion molecules. They are present on plasma membrane of a variety of cell types from primitive metazoans to humans. In the past several years, it has become clear that in addition to providing mechanical adhesion between cells, cadherins play integral roles in tissue morphogenesis and homeostasis. The cadherin family is composed of more than 100 members and classified into several subfamilies, including classical cadherins and protocadherins. Several of these cadherin family members have been implicated in various aspects of neuronal development and function. (1-3) The classical cadherins are associated with a group of cytosolic proteins, collectively called the catenins. While the functional roles of the cadherin-catenin cell adhesion complex have been extensively investigated in epithelial cells, it is now clear that components of the complex are well expressed in central neurons at different stages during development. (4,5) Recent exciting studies have shed some light on the functional roles of cadherins and catenins in central neurons. In this review, we will provide a brief overview of the cadherin superfamily, describe cadherin family members expressed in central neurons, cadherin-catenin complexes in central neurons and then focus on role of the cadherin-catenin complex in dendrite morphogenesis and synapse morphogenesis, function and plasticity. The final section is dedicated to discussion of the emerging list of neural disorders linked to cadherins and catenins. While the roles of cadherins and catenins have been examined in several different types of neurons, the focus of this review is their role in mammalian central neurons, particularly those of the cortex and hippocampus. Accompanying this review is a series of excellent reviews targeting the roles of cadherins and protocadherins in other aspects of neural development.


Treatment of Subarachnoid Hemorrhage-associated Delayed Cerebral Ischemia With Milrinone: A Review and Proposal.
Delayed cerebral ischemia (DCI) following aneurysmal subarachnoid hemorrhage continues to be associated with high levels of morbidity and mortality. This complication had long been thought to occur secondary to severe cerebral vasospasm, but expert opinion now favors a multifactorial etiology, opening the possibility of new therapies. To date, no definitive treatment option for DCI has been recommended as standard of care, highlighting a need for further research into potential therapies. Milrinone has been identified as a promising therapeutic agent for DCI, possessing a mechanism of action for the reversal of cerebral vasospasm as well as potentially anti-inflammatory effects to treat the underlying etiology of DCI. Intra-arterial and intravenous administration of milrinone has been evaluated for the treatment of DCI in single-center case series and cohorts and appears safe and associated with improved clinical outcomes. Recent results have also brought attention to the potential outcome benefits of early, more aggressive dosing and titration of milrinone. Limitations exist within the available data, however, and questions remain about the generalizability of results across a broader spectrum of patients suffering from DCI. The development of a standardized protocol for milrinone use in DCI, specifically addressing areas requiring further clarification, is needed. Data generated from a standardized protocol may provide the impetus for a multicenter, randomized control trial. We review the current literature on milrinone for the treatment of DCI and propose a preliminary standardized protocol for further evaluation of both safety and efficacy of milrinone.


Effects of Cry1Ab transgenic maize on lifecycle and biomarker responses of the earthworm, Eisenia andrei.
A 28-day study was conducted to determine the effects of the Bacillus thuringiensis Cry1Ab toxin on the earthworm Eisenia andrei. Previously, investigations have been limited to life-cycle level effects of this protein on earthworms, and mostly on E. fetida. In this study several endpoints were compared which included biomass changes, cocoon production, hatching success, a cellular metal-stress biomarker (Neutral Red Retention Time; NRRT) and potential genotoxic effects in terms of Randomly Amplified Polymorphic DNA sequences (RAPDs). NRRT results indicated no differences between treatments (p > 0.36), and NRRT remained the same for both treatments at different times during the experiment (p = 0.18). Likewise, no significant differences were found for cocoon production (p = 0.32) or hatching success (p = 0.29). Conversely, biomass data indicated a significant difference between the control treatment and the Bt treatment from the second week onwards (p < 0.001), with the Bt treatment losing significantly more weight than the isoline treatment. Possible confounding factors were identified that might have affected the differences in weight loss between groups. From the RAPD profiles no conclusive data were obtained that could link observed genetic variation to exposure of E. andrei to Cry1Ab proteins produced by Bt maize.


A pyrene-modified cobalt salophen complex immobilized on multiwalled carbon nanotubes acting as a precursor for efficient electrocatalytic water oxidation.
Immobilization of earth-abundant water oxidation catalysts (WOCs) on carbon supports to produce functional electrodes for electrochemical water splitting is a crucial approach for future clean energy production. Herein we report the non-covalent immobilization of a pyrene-bearing cobalt(ii) Schiff base complex (2) on the surface of multiwalled carbon nanotubes (MWCNTs) to form a hybrid anode for electrocatalytic water oxidation. The 2/MWCNT anode displayed excellent catalytic activity and durability in neutral aqueous solution, and a catalytic current density of 1.0 mA cm-2 was achieved at 1.15 V vs. the normal hydrogen electrode (NHE), corresponding to a low overpotential of 330 mV. A Tafel slope of 96 mV per decade was obtained. The Faradaic efficiency of oxygen evolution was more than 90% by bulk electrolysis measurement. After bulk electrolysis, the hybrid anode characterization using X-ray photoelectron spectroscopy (XPS) confirmed that complex 2 decomposed to form heterogeneous cobalt hydroxides and the cobalt hydroxides should be true catalytic active species, which are responsible for electrocatalytic oxygen evolution.


[Kinetics of the ATP-activated ionic currents in the sensory neurons of the rat].
It is shown that ATP being applied to sensory neurons of the rat activates ionic currents with variable kinetics of desensitization. The rate constant of the most "rapid" component of these currents is about 30 ms. The rate constant of the most "slow" component is about 30 s. Pharmacology and ionic properties of different components of the ATP-activated current is, probably, identical.


Proteomic analysis reveals age-related changes in tendon matrix composition, with age- and injury-specific matrix fragmentation.
Energy storing tendons, such as the human Achilles and equine superficial digital flexor tendon (SDFT), are highly prone to injury, the incidence of which increases with aging. The cellular and molecular mechanisms that result in increased injury in aged tendons are not well established but are thought to result in altered matrix turnover. However, little attempt has been made to fully characterize the tendon proteome nor determine how the abundance of specific tendon proteins changes with aging and/or injury. The aim of this study was, therefore, to assess the protein profile of normal SDFTs from young and old horses using label-free relative quantification to identify differentially abundant proteins and peptide fragments between age groups. The protein profile of injured SDFTs from young and old horses was also assessed. The results demonstrate distinct proteomic profiles in young and old tendon, with alterations in the levels of proteins involved in matrix organization and regulation of cell tension. Furthermore, we identified several new peptide fragments (neopeptides) present in aged tendons, suggesting that there are age-specific cleavage patterns within the SDFT. Proteomic profile also differed between young and old injured tendon, with a greater number of neopeptides identified in young injured tendon. This study has increased the knowledge of molecular events associated with tendon aging and injury, suggesting that maintenance and repair of tendon tissue may be reduced in aged individuals and may help to explain why the risk of injury increases with aging.


Recombinant long-acting glycoPEGylated factor IX in hemophilia B: a multinational randomized phase 3 trial.
This multinational, randomized, single-blind trial investigated the safety and efficacy of nonacog beta pegol, a recombinant glycoPEGylated factor IX (FIX) with extended half-life, in 74 previously treated patients with hemophilia B (FIX activity ≤2 IU/dL). Patients received prophylaxis for 52 weeks, randomized to either 10 IU/kg or 40 IU/kg once weekly or to on-demand treatment of 28 weeks. No patients developed inhibitors, and no safety concerns were identified. Three hundred forty-five bleeding episodes were treated, with an estimated success rate of 92.2%. The median annualized bleeding rates (ABRs) were 1.04 in the 40 IU/kg prophylaxis group, 2.93 in the 10 IU/kg prophylaxis group, and 15.58 in the on-demand treatment group. In the 40 IU/kg group, 10 (66.7%) of 15 patients experienced no bleeding episodes into target joints compared with 1 (7.7%) of 13 patients in the 10 IU/kg group. Health-related quality of life (HR-QoL) assessed with the EuroQoL-5 Dimensions visual analog scale score improved from a median of 75 to 90 in the 40 IU/kg prophylaxis group. Nonacog beta pegol was well tolerated and efficacious for the treatment of bleeding episodes and was associated with low ABRs in patients receiving prophylaxis. Once-weekly prophylaxis with 40 IU/kg resolved target joint bleeds in 66.7% of the affected patients and improved HR-QoL. This trial was registered at www.clinicaltrials.gov as #NCT01333111.


Helicopter trimming and tracking control using direct neural dynamic programming.
This paper advances a neural-network-based approximate dynamic programming control mechanism that can be applied to complex control problems such as helicopter flight control design. Based on direct neural dynamic programming (DNDP), an approximate dynamic programming methodology, the control system is tailored to learn to maneuver a helicopter. The paper consists of a comprehensive treatise of this DNDP-based tracking control framework and extensive simulation studies for an Apache helicopter. A trim network is developed and seamlessly integrated into the neural dynamic programming (NDP) controller as part of a baseline structure for controlling complex nonlinear systems such as a helicopter. Design robustness is addressed by performing simulations under various disturbance conditions. All designs are tested using FLYRT, a sophisticated industrial scale nonlinear validated model of the Apache helicopter. This is probably the first time that an approximate dynamic programming methodology has been systematically applied to, and evaluated on, a complex, continuous state, multiple-input multiple-output nonlinear system with uncertainty. Though illustrated for helicopters, the DNDP control system framework should be applicable to general purpose tracking control.


A low-sodium diet supplemented with fish oil lowers blood pressure in the elderly.
To examine effects of dietary fish oil supplementation with sodium restriction on blood pressure in the elderly. In a double-blind dietary intervention lasting 4 weeks, parallel comparisons of blood pressure were made in volunteers assigned to one of four treatment groups: fish oil and low sodium; fish oil and normal sodium; sunflower oil and low sodium; or sunflower oil and normal sodium. Subjects lived at home and attended our nutrition research clinic at fortnightly intervals for dietary counselling and blood pressure measurement. Health volunteers aged 60-80 years were sought by advertisement. A total of 114 men and women were enrolled in two cohorts; 106, with an initial mean blood pressure of 132/77 mmHg, satisfactorily completed the study. All subjects adopted a low-sodium diet and dietary changes were effected by double-blind administration of slow-release sodium chloride or placebo tablets, along with capsules containing either fish or sunflower oil. The primary measure was the within-subject change in blood pressure after 4 weeks of intervention in each dietary treatment group. Urinary sodium excretion in subjects on low-sodium diets decreased whilst potassium excretion was unaffected. Systolic blood pressure (SBP) fell in the group taking sunflower oil with low sodium, but there was only a transient fall in diastolic blood pressure (DBP). In those taking fish oil with normal sodium, the change in blood pressure was not significant, except after adjustment for initial blood pressure and weight changes. When fish oil was combined with low sodium, however, both SBP and DBP were substantially reduced; the reduction in DBP was significantly greater than in the other treatment groups. Dietary fish oil and sodium restriction can interact to lower DBP in the elderly.


Transcription boundaries of U1 small nuclear RNA.
Transcription-proximal stages of U1 small nuclear RNA biosynthesis were studied by 32P labeling of nascent chains in isolated HeLa cell nuclei. Labeled RNA was hybridized to nitrocellulose-immobilized, single-stranded M13 DNA clones corresponding to regions within or flanking a human U1 RNA gene. Transcription of U1 RNA was inhibited by greater than 95% by alpha-amanitin at 1 microgram/ml, consistent with previous evidence that it is synthesized by RNA polymerase II. No hybridization to DNA immediately adjacent to the 5' end of mature U1 RNA (-6 to -105 nucleotides) was detected, indicating that, like all studied polymerase II initiation, transcription of U1 RNA starts at or very near the cap site. However, in contrast to previously described transcription units for mRNA, in which equimolar transcription occurs for hundreds or thousands of nucleotides beyond the mature 3' end of the mRNA, labeled U1 RNA hybridization dropped off sharply within a very short region (approximately 60 nucleotides) immediately downstream from the 3' end of mature U1 RNA. Also in contrast to pre-mRNA, which is assembled into ribonucleoprotein (RNP) particles while still nascent RNA chains, the U1 RNA transcribed in isolated nuclei did not form RNP complexes by the criterion of reaction with a monoclonal antibody for the small nuclear RNP Sm proteins. This suggests that, unlike pre-mRNA-RNP particle formation, U1 small nuclear RNP assembly does not occur until after the completion of transcription. These results show that, despite their common synthesis by RNA polymerase II, mRNA and U1 small nuclear RNA differ markedly both in their extents of 3' processing and their temporal patterns of RNP assembly.


A cell-specific enhancer that specifies lin-3 expression in the C. elegans anchor cell for vulval development.
During C. elegans vulval development, the anchor cell (AC) in the somatic gonad expresses lin-3, activating the EGF receptor signaling pathway in vulval precursor cells (VPCs) and thereby inducing and patterning VPCs. Previous studies with lin-3 mutants and transgene expression have revealed that the level of LIN-3 in the AC must be precisely regulated for proper vulval development. To understand how lin-3 expression is achieved in the AC, we identified a 59 bp lin-3 enhancer sufficient to activate lin-3 transcription solely in the AC. The enhancer contains two E-box elements, and one FTZ-F1 nuclear hormone receptor (NHR) binding site that is mutated in a vulvaless mutant, lin-3(e1417). Mutagenesis studies show that both E-boxes and the NHR binding site are necessary to express lin-3 in the AC. In vitro DNA-binding studies and in vivo functional assays indicate that distinct trans-acting factors, including the E-protein/Daughterless homolog HLH-2 and unidentified nuclear hormone receptor(s), are necessary for lin-3 transcription in the AC and thus are involved in vulval development.


Computer anxiety among nursing students, educators, staff, and administrators.
This article reports the results of a study of computer anxiety among a large (n = 638), diverse sample of nurses (undergraduate and graduate students, educators, staff nurses, and nurse managers). Data were collected using the Oetting Computer Anxiety Scale (COMPAS). Although there was significant variability within and between groups, most nurses exhibited only "mild" computer anxiety and expected to be using computers within three years. Correlates of computer anxiety are also discussed.


The effects of tool scarcity on group climate and affective meaning within the context of a stenciling activity.
Occupational therapists conducting activity analyses often consider the effects of supplies and tools on group process. This study compared three levels of tool supply within the context of a stenciling activity: high scarcity (one essential tool per three subjects), moderate scarcity (two essential tools per three subjects), and no scarcity (three essential tools per three subjects). Thirty-six female subjects rated the activity using Osgood's short-form semantic differential and MacKenzie's Group Climate Questionnaire-Short Form. The data supported the hypothesis of higher group engagement scores in the moderate level of supply than in the high-scarcity or no-scarcity conditions. Though low in all conditions, conflict scores were especially low in the moderate scarcity condition. An unexpected result was found: Subjects experiencing high scarcity had the shortest completion time. No differences between the three supply levels in terms of affective meaning were found. The findings are discussed in terms of the literature of small groups and occupational therapy theory.


Factors associated with immediate relapse among Bahraini heroin abusers.
Demographic characteristics and factors associated with immediate relapse to heroin use among 40 male Bahraini heroin abusers were studied 1 week after discharge from the Drug and Alcohol Rehabilitation Unit in Bahrain. The mean age of the patients was 32.7 years, the age at which drug abuse began ranged from 12 years to 31 years, and the age range of regular use was 15-37 years. More than half the patients were single, unemployed, unskilled labourers with secondary-school education. The vast majority used heroin intravenously. Negative emotional states and drug-related cues were seen by the majority of the subjects as influential in their immediate relapse after discharge. Findings suggest that the treatment and rehabilitation unit in Bahrain should look into the issues of after-care.


Discriminating micropathogen lineages and their reticulate evolution through graph theory-based network analysis: the case of Trypanosoma cruzi, the agent of Chagas disease.
Micropathogens (viruses, bacteria, fungi, parasitic protozoa) share a common trait, which is partial clonality, with wide variance in the respective influence of clonality and sexual recombination on the dynamics and evolution of taxa. The discrimination of distinct lineages and the reconstruction of their phylogenetic history are key information to infer their biomedical properties. However, the phylogenetic picture is often clouded by occasional events of recombination across divergent lineages, limiting the relevance of classical phylogenetic analysis and dichotomic trees. We have applied a network analysis based on graph theory to illustrate the relationships among genotypes of Trypanosoma cruzi, the parasitic protozoan responsible for Chagas disease, to identify major lineages and to unravel their past history of divergence and possible recombination events. At the scale of T. cruzi subspecific diversity, graph theory-based networks applied to 22 isoenzyme loci (262 distinct Multi-Locus-Enzyme-Electrophoresis -MLEE) and 19 microsatellite loci (66 Multi-Locus-Genotypes -MLG) fully confirms the high clustering of genotypes into major lineages or "near-clades". The release of the dichotomic constraint associated with phylogenetic reconstruction usually applied to Multilocus data allows identifying putative hybrids and their parental lineages. Reticulate topology suggests a slightly different history for some of the main "near-clades", and a possibly more complex origin for the putative hybrids than hitherto proposed. Finally the sub-network of the near-clade T. cruzi I (28 MLG) shows a clustering subdivision into three differentiated lesser near-clades ("Russian doll pattern"), which confirms the hypothesis recently proposed by other investigators. The present study broadens and clarifies the hypotheses previously obtained from classical markers on the same sets of data, which demonstrates the added value of this approach. This underlines the potential of graph theory-based network analysis for describing the nature and relationships of major pathogens, thereby opening stimulating prospects to unravel the organization, dynamics and history of major micropathogen lineages.


Reduced-intensity hematopoietic cell transplantation in older patients with AML/MDS: umbilical cord blood is a feasible option for patients without HLA-matched sibling donors.
Umbilical cord blood (UCB) has increased access to hematopoietic cell transplantation (HCT) for patients without HLA-matched sibling donors (MSD). We compared outcomes of HCT using MSD (N=38) or UCB (N=60) among older patients (age ≥ 55 years) with AML or myelodysplastic syndromes (MDS). All patients received a reduced intensity regimen consisting of CY, fludarabine and 200 cGy TBI. Median age at HCT was 63 years for MSD and 61 years for UCB recipients. Among UCB recipients, 95% received two UCB units and 88% received 1-2 locus HLA-mismatched units to optimize cell dose. OS at 3-years was 37% for MSD and 31% for UCB recipients (P=0.21). On multivariate analysis, donor source (MSD vs UCB) did not impact risks of OS, leukemia-free survival and relapse or treatment-related mortality. UCB is feasible as an alternative donor source for reduced-intensity conditioning HCT among older patients with AML and MDS who do not have a suitable MSD.


The decline of adult smallpox in eighteenth-century London.
Smallpox was probably the single most lethal disease in eighteenth-century Britain, but was a minor cause of death by the mid-nineteenth century. Although vaccination was crucial to the decline of smallpox, especially in urban areas, from the beginning of the nineteenth century, it remains disputed the extent to which smallpox mortality declined before vaccination. Analysis of age-specific changes in smallpox burials within the large west London parish of St Martin-in-the-Fields revealed a precipitous reduction in adult smallpox risk from the 1770s, and this pattern was duplicated in the east London parish of St Dunstan's. Most adult smallpox victims were rural migrants, and such a drop in their susceptibility is consistent with a sudden increase in exposure to smallpox in rural areas. We investigated whether this was due to the spread of inoculation, or an increase in smallpox transmission, using changes in the age patterns of child smallpox burials. Smallpox mortality rose among infants, and smallpox burials became concentrated at the youngest ages, suggesting a sudden increase in infectiousness of the smallpox virus. Such a change intensified the process of smallpox endemicization in the English population, but also made cities substantially safer for young adult migrants.


The Increased Expression of CCL20 and CCR6 in Rectal Mucosa Correlated to Severe Inflammation in Pediatric Ulcerative Colitis.
Background/Aims. The aim of this study is to clarify the differences of CCL20 and CCR6 expression, chemokine correlated to intestinal homeostasis, between pediatric and adult ulcerative colitis (UC) patients. Methods. Onehundred forty-one patients who underwent proctocolectomy were divided to two groups including childhood-onset UC (CUC, <16 years old, n = 24) and adult-onset UC (AUC, ≧16 years old, n = 117). A total of 141 formalin-fixed, paraffin-embedded tissue samples of rectum were obtained from these patients. Histological inflammation of rectum in resected specimen was evaluated by using Geboes histological assessment. In immunohistochemistry study, the CCL20 expression was evaluated by intensity and the stained area, and the CCR6 expression was evaluated by lymphocytes infiltration pattern. Results. CCL20 score and CCR6 positive lymphocytes infiltration pattern were statistically significantly correlated with histological inflammation severity of UC in all patients (P < 0.05). CCL20 and CCR6 expression in CUC were statistically significantly higher than that in AUC in all or pathologically severe cases (P < 0.05). Conclusions. CCL20 and CCR6 may play a significant role in local damage and pathological changes in UC especially pediatric patients. In the future, our understanding of the differences in CCL-CCR6 interaction between adults and children may lead to the pathogenesis of IBD.


Cervical Spine Deformity-Part 3: Posterior Techniques, Clinical Outcome, and Complications.
The goals of cervical deformity surgery include deformity correction, restoration of horizontal gaze, decompression of neural elements, spinal stabilization with a biomechanically sound construct, and meticulous arthrodesis technique to prevent pseudoarthrosis and minimizing surgical complications. Many different surgical options exist, but selecting the correct approach that ensures the optimal clinical outcome can be challenging and often controversial. In this last part of the cervical deformity review series, various posterior deformity correction techniques are discussed in detail, along with an overview of surgical outcome and postoperative complications.


Novel Organic Mineral Complex Prevents High-Fat Diet-Induced Changes in the Gut and Liver of Male Sprague-Dawley Rats.
Diet-induced obesity and metabolic syndrome are associated with the onset of gastrointestinal diseases, such as hepatic steatosis and gut inflammation. Prior research shows that a proprietary soil-derived organic mineral complex (OMC) prevents hyperglycemia, endotoxemia, and liver injury in rats fed a high-fat diet (HFD) for 10 weeks. The aim of this study was to further examine the effects of OMC on the liver and gastrointestinal health of these rats. Six-week-old male Sprague-Dawley rats (n = 36) were divided into two dietary groups: Chow or HFD fed for 10 weeks. Animals were further divided (n = 6/group) and administered 0, 0.6, or 3.0 mg/mL OMC in their drinking water. The 10-week HFD resulted in significant liver fat accumulation. Both OMC doses prevented hepatic increases in the glycation end product Nε-(carboxymethyl)lysine (CML) induced by HFD (p < 0.05). Low-dose OMC was associated with higher expression of occludin in the small intestine of rats fed either diet (two-way ANOVA, p < 0.042). Linear discriminant analysis (LDA) effect size (LEfSe) indicated significant differences in fecal microbial composition of untreated HFD-fed rats in comparison to untreated Chow rats at 10 weeks (LDA score > 2.0 : 18). After 10 weeks, untreated HFD-fed rats were also more abundant in bacteria associated with obesity and metabolic disease in comparison to corresponding week 0 samples (LDA score > 2.0 : 31), 10-week untreated Chow (LDA > 2.0 : 18), or 10-week OMC-treated HFD-fed rats (0.6 mg/mL; LDA > 2.0 : 80, 3.0 mg/mL; LDA > 2.0 : 8). Low-dose OMC prevented the HFD-induced increase in the Firmicutes-to-Bacteroidetes (F/B) ratio (p < 0.0416). Study animals treated with OMC exhibited no significant changes in the gut microbiota at week 10, although gut inflammatory biomarkers were not significantly altered by diet or OMC treatment. These results indicate that OMC supplementation ameliorates glycosylation reactions and modifies HFD-induced alterations in the intestinal microbiota.


[Association of Doppler flowmetry with perinatal outcome in patients with oligohydramnios].
Oligohydramnios is sometimes associated with poor perinatal outcome. Our aim was to determine the association of Doppler flowmetry with perinatal outcome in patients with oligohydramnios. We carried out a prospective cross-sectional study that included 130 patients with pregnancy between 30 to 41 weeks of gestation; we compared the measurement of the resistance index in the umbilical artery by Doppler flowmetry and the measurement of amniotic fluid in patients with oligohydramnios. A follow-up of patients after birth was made, recording the Apgar score and whether they required special attention. The data were analyzed using descriptive statistics and we calculated the sensitivity, specificity, accuracy, and predictive values of the tests. Sensitivity, specificity and accuracy were obtained for oligohydramnios associated to perinatal death and for five-minute Apgar, with results of 100, 0, 50 % and 0, 0, 0 % respectively. The resistance index of umbilical artery related to perinatal death had sensitivity, specificity and accuracy of 100, 96 and 98 %, while in the Apgar were 0, 95 and 47.5 %, respectively. Doppler flowmetry of the umbilical artery showed a higher sensitivity, specificity and accuracy to diagnose perinatal death than the measurement of oligohydramnios and the Apgar score.


Effects of a biomimetic superoxide dismutase on complete and multistage carcinogenesis in mouse skin.
The effects of a selective detoxifier of the proximate oxygen radical, superoxide anion, on the induction of tumors in the skin of CD-1 mice by either the initiation-promotion regimen or the complete carcinogenesis process were investigated. The principle agent of interest, copper (II) (3,5-diisopropylsalicylate)2 (CuDIPS), is a low molecular weight, lipophilic copper coordination complex that catalytically disproportionates superoxide anion at a rate comparable to native Cu-Zn superoxide dismutase (SOD). The protocols used to elicit tumors were: (i) a single application of 0.2 mumol of 7,12-dimethylbenz[a]anthracene (DMBA) followed by twice-weekly applications of 12-O-tetradecanoylphorbol-13-acetate (TPA) in an initiation-promotion study, and (ii) either a single application of 3.6 mumol DMBA followed by no further treatment or weekly applications of 0.2 mumol DMBA in complete carcinogenesis protocols. Application of 2 mumol CuDIPS 15 min prior to the initiating dose of DMBA was without significant effect on tumor yield or incidence, whereas application prior to each dose of TPA substantially reduced tumor incidence and yield. This anti-promoting property of CuDIPS can be attributed to its SOD-mimetic activity in as much as the corresponding zinc coordination complex lacking in SOD activity, zinc (II) (3,5-diisopropylsalicylate)2, was non-inhibitory. Significant reductions in tumor yield were also observed when CuDIPS was applied prior to DMBA in either of the complete carcinogenesis protocols. Additionally, covalent binding of [3H]DMBA to epidermal DNA was markedly reduced by CuDIPS pre-treatment, suggesting that the anti-carcinogenic properties may reflect a perturbation in superoxide anion-dependent metabolic activation of DMBA. The induction by DMBA of ornithine decarboxylase activity, a biochemical marker of tumor promoter activity, was not affected by CuDIPS; however, induction of ornithine decarboxylase by TPA was potently blocked. Collectively, these effects of a biomimetic SOD further implicate reactive oxygen species at multiple stages in chemical carcinogenesis.


Clinical Efficacy and Safety of Nivolumab in Malignant Non-Pleural Mesothelioma: A Multicenter, Open-Label, Single-Arm, Japanese Phase II Trial (Viola) Protocol.
There is no authorized treatment for malignant non-pleural mesothelioma (MNPM) worldwide. In contrast to malignant pleural mesothelioma, MNPM has not been investigated, and no treatment has been established due to its rarity. This multicenter, open-label, single-arm, Japanese phase II trial aims at evaluating the efficacy and safety of nivolumab, an immune checkpoint inhibitor, in advanced or metastatic MNPM treatment. This phase II trial commenced in October 2020. Twenty-three patients with advanced or metastatic MNPM who meet the inclusion and exclusion criteria were enrolled from five institutions within 2 years. Regardless of prior therapy, 240 mg of nivolumab will be administered intravenously to MNPM patients every 2 weeks to investigate its efficacy and safety until disease progression or unacceptable toxicities are detected, or the patient's condition meets the withdrawal criteria. The primary endpoint is the objective response rate by central assessment following the Response Evaluation Criteria in Solid Tumors version 1.1. The secondary endpoints include disease control rate, overall survival, progression-free survival, adverse events, and treatment-related adverse events. This is the first prospective investigator-initiated trial to evaluate the effect of nivolumab monotherapy for MNPM.


A novel bone conduction implant (BCI): engineering aspects and pre-clinical studies.
Percutaneous bone anchored hearing aids (BAHA) are today an important rehabilitation alternative for patients suffering from conductive or mixed hearing loss. Despite their success they are associated with drawbacks such as skin infections, accidental or spontaneous loss of the bone implant, and patient refusal for treatment due to stigma. A novel bone conduction implant (BCI) system has been proposed as an alternative to the BAHA system because it leaves the skin intact. Such a BCI system has now been developed and the encapsulated transducer uses a non-screw attachment to a hollow recess of the lateral portion of the temporal bone. The aim of this study is to describe the basic engineering principals and some preclinical results obtained with the new BCI system. Laser Doppler vibrometer measurements on three cadaver heads show that the new BCI system produces 0-10 dB higher maximum output acceleration level at the ipsilateral promontory relative to conventional ear-level BAHA at speech frequencies. At the contralateral promontory the maximum output acceleration level was considerably lower for the BCI than for the BAHA.


The problem of occlusal caries and its current management.
Occlusal caries has become relatively more important, due to the changing pattern of dental caries that has occurred in the developed countries of the world. Accurate diagnosis of the activity of a carious lesion is important, because arrest and remineralization are possible. Diagnosis of occlusal caries is difficult due to the morphology of the occlusal surface. Unfortunately, topical action of fluorides is thought to be making diagnosis increasingly difficult. The literature suggests that we should be examining for caries under good lighting and a dry field, the use of a blunt probe being reserved for gently removing debris from fissures to enable good visualisation. Bitewing radiography is valuable in the detection of occlusal as well as approximal caries, but good radiographic technique and careful viewing are essential. Electronic caries detection may well prove useful in the future. The traditional restorative approach to occlusal caries resulted in inconsistent treatment decisions, poor durability of restorations, and the unfortunate consequences of replacement restorations. It seems logical to take a more conservative, preventive approach in the management of dental caries. Fissure sealants should be used preventively for the caries-prone patient, and therapeutically for the suspect or early carious lesion. Where the caries has spread into dentine, as shown radiographically, then the sealant restoration may be more suitable. These techniques have been shown to be safe and effective, although careful case selection, appropriate manipulation of materials, recall, and maintenance are essential to long-term success.(ABSTRACT TRUNCATED AT 250 WORDS)


Performance Monitoring Applied to System Supervision.
Nowadays, automation is present in every aspect of our daily life and has some benefits. Nonetheless, empirical data suggest that traditional automation has many negative performance and safety consequences as it changed task performers into task supervisors. In this context, we propose to use recent insights into the anatomical and neurophysiological substrates of action monitoring in humans, to help further characterize performance monitoring during system supervision. Error monitoring is critical for humans to learn from the consequences of their actions. A wide variety of studies have shown that the error monitoring system is involved not only in our own errors, but also in the errors of others. We hypothesize that the neurobiological correlates of the self-performance monitoring activity can be applied to system supervision. At a larger scale, a better understanding of system supervision may allow its negative effects to be anticipated or even countered. This review is divided into three main parts. First, we assess the neurophysiological correlates of self-performance monitoring and their characteristics during error execution. Then, we extend these results to include performance monitoring and error observation of others or of systems. Finally, we provide further directions in the study of system supervision and assess the limits preventing us from studying a well-known phenomenon: the Out-Of-the-Loop (OOL) performance problem.


The central role of EED in the orchestration of polycomb group complexes.
Polycomb repressive complexes 1 and 2 (PRC1 and 2) play a critical role in the epigenetic regulation of transcription during cellular differentiation, stem cell pluripotency and neoplastic progression. Here we show that the polycomb group protein EED, a core component of PRC2, physically interacts with and functions as part of PRC1. Components of PRC1 and PRC2 compete for EED binding. EED functions to recruit PRC1 to H3K27me3 loci and enhances PRC1-mediated H2A ubiquitin E3 ligase activity. Taken together, we suggest an integral role for EED as an epigenetic exchange factor coordinating the activities of PRC1 and 2.


Evaluation of brucellin skin test for bovine brucellosis.
The diagnostic efficiency of the Brucellin skin test for bovine brucellosis was evaluated using cattle of known history. The test was negative in six out of 14 heifers (42.9%) infected with virulent Brucella abortus (Br. abortus) strain 544. In four cattle vaccinated with a reduced dose of Br. abortus strain 19 vaccine (5 X 10(9) live organisms) the skin test became positive in all the animals but two weeks after immunization. However, all the vaccinates became negative 14 weeks after vaccination, whereas nine out of 14 heifers (64.3%) vaccinated with killed Br. abortus 45/20 adjuvant vaccine (Duphavac vaccine) were still positive 18 months post immunization. Four control cattle were persistently negative. It was considered that the procedure would be most useful for testing non-exposed cattle and should be used as a screen test. It is not useful as a diagnostic test for cattle immunized with Duphavac vaccine. A recommendation is made for interpreting the Brucellin skin test for the diagnosis of bovine brucellosis in non-exposed herds.