Novel method for identifying sequence-specific DNA-binding proteins.
We developed a general method for the enrichment and identification of sequence-specific DNA-binding proteins. A well-characterized protein-DNA interaction is used to isolate from crude cellular extracts or fractions thereof proteins which bind to specific DNA sequences; the method is based solely on this binding property of the proteins. The DNA sequence of interest, cloned adjacent to the lac operator DNA segment is incubated with a lac repressor-beta-galactosidase fusion protein which retains full operator and inducer binding properties. The DNA fragment bound to the lac repressor-beta-galactosidase fusion protein is precipitated by the addition of affinity-purified anti-beta-galactosidase immobilized on beads. This forms an affinity matrix for any proteins which might interact specifically with the DNA sequence cloned adjacent to the lac operator. When incubated with cellular extracts in the presence of excess competitor DNA, any protein(s) which specifically binds to the cloned DNA sequence of interest can be cleanly precipitated. When isopropyl-beta-D-thiogalactopyranoside is added, the lac repressor releases the bound DNA, and thus the protein-DNA complex consisting of the specific restriction fragment and any specific binding protein(s) is released, permitting the identification of the protein by standard biochemical techniques. We demonstrate the utility of this method with the lambda repressor, another well-characterized DNA-binding protein, as a model. In addition, with crude preparations of the yeast mitochondrial RNA polymerase, we identified a 70,000-molecular-weight peptide which binds specifically to the promoter region of the yeast mitochondrial 14S rRNA gene.


Long-time-storage mechanism for Tm:YAG in a magnetic field.
We obtained a long-time-storage mechanism for spectral features in thulium ions doped into YAG by applying a magnetic field that splits the electronic ground state. We show experimentally that the storage time can be more than 30 s, which is 3 orders of magnitude longer than that of the metastable state that normally is used for information storage in this material. Level splitting and storage lifetimes for various magnetic field strengths of as much as 5 T were investigated. This storage mechanism will be relevant in the many coherent transient-based signal-processing schemes in which Tm:YAG is being used, and we demonstrate long-time storage in a basic data storage application.


[Reaction of the thyroid gland to a single and prolonged administration of prolactin].
A study was made of the thyroid and hypophysis response to the administration of a summary analogous dose of prolactin injected in a single dose or in the course of 5 days. Experiments were made on female rats at the diestrus stage. The content of thyroid hormones and thyrotropic hormone (TTH) in the blood was determined by radioimmunoassay. The concentration of thyroid hormones and TTH was compared in the basal state and 30, 60, 120 and 180 min after prolactin injection. An acute prolactin administration was shown to result in an elevated TTH level on the 30th minute and T4 on the 60th minute. Five-day hyperprolactinemia caused a decrease in the content of thyroid hormones in the blood and disturbed positive correlative interrelationships between the thyroid hormones and negative ones between thyroxine and TTH. It is assumed that the preservation of a high prolactin level in the blood can maintain the hyperthyroid state.


[Lab-on-a-chip (microfluidics) technology].
Over the last decade, the lab-on-a-chip concept has received considerable attention. This technology promises significant advantages in terms of speed, cost, sample/reagent consumption, contamination, efficiency and automation. In this review, we focus on lab-on-a-chip with fluid flow along micro-scale channels (microfluidics) and its application on life science and biotechnology.


[Intravenous administration of ritodrine to pregnant insulin-dependent diabetics. Metabolic impact].
Seven diabetic patients who had been dependent on insulin before pregnancy (classes B, C, D of White) threatened to go into premature labour between the 31st and 36th week of the pregnancy. They were treated by intravenous administration of Ritodrine, the dosage varying between 100 and 400 micrograms per minute. The changes in the mother consisted in a rise in the blood glucose level which rose from 140 +/- 34 mg % to 212 +/- 56 mg % (p less than 0.01). This occurred in spite of a mean rise of 49% of insulin (p less than 0.0025) and a moderate ketonuria in two patients and a severe acido-ketosis in two further patients. In five patients there were no effects on the fetus as far as repeated observations of the fetal heart rate could detect. On the other hand, in the two mothers who developed acido-ketosis during the transfusion the fetal heart rythm showed signs of acute fetal distress, which however was temporary. Oscillations were reduced and there were late decelerations. These disappeared when the maternal metabolic state was brought back to equilibrium. 13.1 +/- 7.3 days were gained for the fetus by using tocolytic treatment, and this is considered a success. Discussion of these cases is accompanied by a complete review of the literature of the administration of beta-mimetic drugs in diabetic women. The authors point out that special precautions have to be taken and special conditions have to be fulfilled when this type of treatment is used. These consist of changes in the amounts of insulin that are given and particularly careful observation of the fetal heart rhythm before and during treatment.


Measuring actin dynamics in endothelial cells.
Cytoplasmic actin distributes between monomeric and filamentous phases in cells. As cells crawl, actin polymerizes near the plasma membrane of expanding peripheral cytoplasm and depolymerizes elsewhere. Thus, the finite actin filament lifetime, the diffusivity of actin monomer, and the distribution of actin between the polymer and monomer phases are key parameters in cell motility. The dynamics of cellular actin can be determined by following the evolution of fluorescence in the techniques of photoactivated fluorescence (PAF) or fluorescence recovery after photobleaching (FRAP) of microinjected actin derivatives. A mathematical model is discussed that measures monomer diffusion coefficients, filament turnover rates, and the fraction of actin polymerized from measurements of the evolution of fluorescence from a photoactivated band [Tardy et al. (1995) Biophys. J., 69:1674-1682; McGrath et al. (1998) Biophys. J., in press]. Applying this model to subconfluent endothelial cells shows that approximately 40% of the actin is polymer and that these filaments turn over on average every 6 minutes. This report discusses how PAF and FRAP can be combined with more traditional biochemistry to probe actin cytoskeleton remodeling in endothelial cells.


Grain boundaries at the surface of consolidated MgO nanocrystals and acid-base functionality.
The increase of the surface basicity-acidity of MgO material by factors of 1.8-3.0 due to consolidation of its nanocrystals was demonstrated by the indicator titration. It was shown that the parallel increase of surface acidity and basicity is attributed to the formation of grain boundaries (GB) after MgO aerogel densification. A simple model predicting the increase of surface acidity-basicity of MgO that correlates with the results of direct measurements was proposed. The model is based on the study of the fine atomic structure at GB surface areas in consolidated MgO nanocrystals in the framework of Density Functional Theory. It is found that the displacements of coordinatively unsaturated surface ions near the GB are significant at the distances ~3-4 atomic layers from the geometrical contact plane between nanocrystals. The detailed analysis of atomic positions inside GB demonstrated the coordination deficiency of surface atoms at the GB areas leading to the formation of stretched bonds and to creation of low coordinated surface ions due to splitting of coordination numbers of surface atoms belonging to GB areas. Density of states for electrons shows the existence of additional states in the band gap close to the bottom of the conduction band. The adsorption energy of CO2 molecules atop oxygen atoms exposed at surface GB areas is of the same order of magnitude as that reported for oxygen atoms at crystallographic edges and corners of MgO crystals. It provides additional options for bonding of molecules at the surface of nanocrystalline MgO increasing the adsorption capacity and catalytic activity.


Genetic engineering and coagulation factors.
It is unfortunate that we cannot report, in the area of coagulation, advances that have been seen in related fields such as thrombolytic therapy. The reported progress (Gold et al, 1984; Van de Werf et al, 1984) with human recombinant tissue plasminogen activator (Pennica et al, 1983) augers well for the application of recombinant technology to the problems faced by patients with coagulation defects. While plasminogen activator is being assessed in an acute therapeutic setting, its use signals a beginning of the application of the technology to abnormalities of the haemostatic mechanism. Chronic administration of coagulation factors for prophylaxis and replacement therapy would appear to be just one more step down the pathway illuminated by the biochemists, microbiologists and cell biologists who have preceded the clinicians in this promising area. There is no record of the use of genetically engineered materials in the treatment of coagulation defects, primarily because the body of knowledge and refined techniques have only recently been acquired. For this reason we have had to project developments in other areas onto the problems that exist for the haemostatically compromised patient. In describing the potential usefulness of these technologies, it is difficult to ascertain where the logical projection, from a fully investigated model system, diverges from flights of imaginative fancy. Cloning projects considered overly ambitious and grandiose at the beginning of this decade are already accomplished feats. The feasibility of gene therapy in the mammalian system has been demonstrated, and trade publications now discuss governmental approval for investigative use of this procedure in 1985. Panels of physicians, scientists and even politicians now seriously contemplate and promulgate views and regulations pertaining to the efficacy and ethics of the use of genetic engineering in the treatment of human disease. The haemophilias will certainly be among the first genetic diseases to be approached with the techniques of recombinant DNA technology. Diagnostic testing, using cloned DNA, is already underway and therapeutic trials are predicted for the near future. Every observer of this rapidly growing field has to define for himself when the future is. For the potential carrier of haemophilia B, the future is now. For the physician managing the patient with a haemophilic inhibitor, the future can't come soon enough. And, for those who are concerned with man's tampering with the gene pools of living things, from viruses to humans, the future comes too quickly to be dealt with in a rational and understanding way.


Mid- and Longer-term Follow up of Chimney and/or Periscope Grafts and Risk Factors for Failure.
The aim was to report on chimney and periscope grafts (CPGs) and their mid- and longer-term outcomes when they are used to preserve reno-visceral artery (RVA) perfusion in endovascular repair of pararenal (PRAAs) or thoraco-abdominal aortic aneurysm (TAAAs). In addition, factors associated with CPG failure are presented. Limited data exist on the outcomes of CPGs, and mid- and long-term results are generally not reported. This was a prospective study in a cohort of 100 patients with PRAA (69) or TAAA (31). A total of 224 (mean 2.24 per patient) RVAs were preserved with 136 (61%) chimney and 88 (39%) periscope grafts. CPGs were constructed mainly using self expandable stent grafts. Patients were followed by clinical examination, CTA (82%), and/or duplex (18%). Data were collected until February 2015. CPG immediate technical success was 99% (222/224 branches). Mean follow up was 29 months (range 0-65; SD 17); 59% patients were followed > 2 years, 30% > 3 years, and 16% > 4 years. Post-operatively, CPG occlusion was observed early (≤30 days) in three (1.3%) branches and during follow up in 10 (4.5%). At 36 and 48 months, the estimated primary patency was 93% and 93%. After corrective percutaneous (10) or surgical (3) re-interventions, the estimated secondary patency was 96% and 96%. Thirty day mortality was 2%; at 36 and 48 months the estimated patient survival was 79%. Significant shrinkage (72 [SD 23] vs. 62 [SD 24] mm; p < .001) was observed, with a substantial reduction (>5 mm) in 55 patients, and sac enlargement in four. Incomplete aneurysm sac sealing was treated successfully by a secondary intervention in 15 patients. Self expandable CPGs have proved to be a highly successful and durable treatment for RVA preservation up to 5 years. Incomplete CPG expansion, inadequate length, and CPG use in small and diseased target arteries were risk factors for occlusion. These mid- and longer-term results support CPG use to treat PRAAs or TAAAs in patients unfit for open surgery or fenestrated/branched stent grafts.


Elevated carbohydrate antigen 19-9 in patients with true (epithelial) splenic cysts--Rare or undiscovered?
Carbohydrate antigen 19-9 is a well known marker for pancreatic adenocarcinoma. However, its limitation is its nonspecificity, because elevated levels may be encountered in other gastrointestinal disorders, both benign and malignant. The following case is a patient with a true (epithelial) splenic cyst with elevated serum levels of carbohydrate antigen 19-9.


Pre-adolescents with multiple health complaints redeem more prescriptions: A follow-up study in the Danish National Birth Cohort.
Aims: This study aimed to examine the association between multiple health complaints (MHC) in pre-adolescence and prescription redemption in adolescence. Methods: This was a nationwide population-based study based on the Danish National Birth Cohort for an average of 6.9 years (2010-2018). A total of 96,382 children were invited at the age of 11. A modified version of the Health Behaviour in School Children Symptom Checklist was dichotomised into the World Health Organization's definition of MHC (⩾2 complaints, each with a frequency of at least weekly, yes/no). The number of prescriptions was retrieved from Danish registries. Negative binomial regression estimated incidence rate ratios (IRRs) comparing children with MHC to children without. Prescription redemption was further stratified by psychiatric/somatic medication and into subtypes of prescriptions. Results: A total of 47,365 (49.1%) children participated (Mage=11.2 years, 52% girls). MHC were reported by 10.3%. The unadjusted IRR (MHC vs. no MHC) of all types of redemptions was 1.57 (95% confidence interval (CI) 1.49-1.64). Results were robust to adjustment for socio-demographic variables and somatic/psychiatric morbidity at baseline (IRR=1.47; 95% CI 1.40-1.54). Associations were especially strong for psychiatric medication (adjusted IRR=3.88; 95% CI 3.43-4.40) and were modified by neither sex nor maternal education. Conclusions: MHC in pre-adolescents predict prescription redemption. This implies that changes in MHC might be indicative of changes in public health. This requires further study, as the cause of a change in reporting of symptoms might also cause a change in treatment response. The latter determines whether prescriptions are treating ill-being or needlessly medicalising subjective symptoms.


Impact of an Antenatal Counseling on Use of Modern Family Planning Methods in the Postpartum in Rural Guinea.
In Guinea, family planning (FP) uptake remains low. The objective of this study was to compare the impact of two types of antenatal counseling on modern FP uptake in the postpartum in rural Guinea. This was a two-group non-equivalent study comparing the impact of a reinforced antenatal counseling (intervention) to the routine antenatal counseling (control). The study included 404 pregnant women at five rural health centres in Forécariah district, Western Guinea. Each woman was followed up until the ninth month postpartum. The study was conducted from October 12, 2013 to December 30, 2014. Findings showed that at the ninth month postpartum, use of modern FP was significantly higher in the intervention group than in the control group (5.7% and 1.1%, respectively; p=0.024). However, 67.6% and 65.7% of women in the intervention group and the control group, respectively, abstained from sexual intercourse at the sixth month postpartum and had the intention to do so until the child walks. At the ninth month postpartum such women represented 70.5% and 59.5%, respectively. Therefore, a longer study period is recommended to assess the effect of antenatal counseling on use of modern FP in the postpartum in Guinea.


Extracellular Vesicles as Unique Signaling Messengers: Role in Lung Diseases.
Extracellular vesicles (EVs) are lipid bilayer-enclosed extracellular particles carrying rich cargo such as proteins, lipids, and microRNAs with distinct characteristics of their parental cells. EVs are emerging as an important form of cellular communication with the ability to selectively deliver a kit of directional instructions to nearby or distant cells to modulate their functions and phenotypes. According to their biogenesis, EVs can be divided into two groups: those of endocytic origin are called exosomes and those derived from outward budding of the plasma membrane are called microvesicles (also known as ectosomes or microparticles). Under physiological conditions, EVs are actively involved in maintenance of pulmonary hemostasis. However, EVs can contribute to the pathogenesis of diseases such as chronic obstructive pulmonary disease, asthma, acute lung injury/acute respiratory distress syndrome, interstitial lung disease, and pulmonary arterial hypertension. EVs, especially those derived from mesenchymal/stromal stem cells, can also be beneficial and can curb the development of lung diseases. Novel technologies are continuously being developed to minimize the undesirable effects of EVs and also to engineer EVs so that they may have beneficial effects and can be used as therapeutic agents in lung diseases. © 2021 American Physiological Society. Compr Physiol 11:1351-1369, 2021.


Quality of maternal and paternal care predicts later stress reactivity in the cooperatively-breeding marmoset (Callithrix geoffroyi).
Variation in the early postnatal social environment can have lasting effects on hypothalamic-pituitary-adrenal (HPA) axis stress responses. Both rats and macaque monkeys subjected to low quality or abusive maternal care during the early postnatal period have more pronounced HPA responses to environmental stressors throughout development and into adulthood compared to animals reared in higher quality early maternal environments. However, little is known about the relative contributions to HPA stress response styles in developing offspring in species in which offspring care is routinely provided by group members other than the mother, such as in cooperatively breeding mammals. Marmoset monkeys exhibit cooperative offspring rearing, with fathers and older siblings providing care in addition to that provided by the mother. We evaluated the effects of early maternal, paternal, and older sibling care on HPA responses to social separation across development in captive white-faced marmoset offspring (Callithrix geoffroyi). We monitored offspring care by mothers, fathers, and older siblings in marmosets for the first 60 days of life. Later in development, each marmoset experienced three standardized social separation/novelty exposure stressors at 6, 12, and 18 months of age. During separation, we collected urine samples and analyzed them via enzyme immunoassay for cortisol levels. Infants that received higher rates of rejections from the entire family group showed higher cortisol responses to social separation. This relationship was found when mothers, fathers, and older siblings, were analyzed separately as well. No differences in cortisol responses were found between offspring that received high and low rates of carrying or high and low rates of licking and grooming by any group member. In the cooperatively breeding marmoset, early social cues from multiple classes of caregivers may influence HPA stress responses throughout the lifespan.


Positron emission tomography in non-small-cell lung cancer: prediction of response to chemotherapy by quantitative assessment of glucose use.
To prospectively evaluate the use of positron emission tomography with the glucose analog fluorodeoxyglucose (FDG-PET) to predict response to chemotherapy in patients with advanced non-small-cell lung cancer (NSCLC). Patients with stage IIIB or IV NSCLC scheduled to undergo platinum-based chemotherapy were eligible for this study. Patients were studied by FDG-PET before and after the first cycle of therapy. Based on previous studies, a reduction of tumor FDG uptake by more than 20% as assessed by standardized uptake values (SUV) was used as a criterion for a metabolic response. Furthermore, changes in tumor SUVs were compared with changes in FDG net-influx constants (Ki) and tumor/muscle ratios (t/m). Fifty-seven patients were included in the study. There was a close correlation between metabolic response and best response to therapy according to Response Evaluation Criteria in Solid Tumors (P <.0001; sensitivity and specificity for prediction of best response, 95% and 74%, respectively). Median time to progression and overall survival were significantly longer for metabolic responders than for metabolic nonresponders (163 v 54 days and 252 days v 151 days, respectively). Similar results were obtained when Ki was used to assess tumor glucose use, whereas changes in t/m showed considerable overlap between responding and nonresponding tumors. In NSCLC, reduction of metabolic activity after one cycle of chemotherapy is closely correlated with final outcome of therapy. Using metabolic response as an end point may shorten the duration of phase II studies evaluating new cytotoxic drugs and may decrease the morbidity and costs of therapy in nonresponding patients.


Effects of B0 eddy currents on imaging isocenter shifts in 0.35-T MRI-guided radiotherapy (MR-IGRT) system.
The purpose of this study was to measure gantry angle-related eddy currents in a 0.35-T MRI-Linac and determine if B0 (zeroth order) eddy currents are the primary cause of gantry angle-dependent imaging isocenter shifts vs other potential causes like B0 inhomogeneities and gradient (first order) eddy currents. For conventional Cartesian acquisitions, B0 eddy currents can cause imaging isocenter shifts along both phase encode and readout directions. Gradient eddy currents can cause spatial distortion along both the phase encode and readout directions. Center frequency offsets can cause imaging isocenter shifts along the readout direction that vary with readout gradient polarity. MRI-related eddy currents and imaging isocenter shifts were measured on a 0.35-T MRI-Linac at gantry angles from 0° to 330° in increments of 30° . All measurements were made after gradient shimming and center frequency tuning at each planned gantry angle. Eddy current and field homogeneity measurements were conducted using a 24-cm diameter spherical phantom. Gradient and B0 eddy currents were calculated from the free induction decays (FIDs) resulting from selective excitation of slices located ±5 cm from isocenter. B0 eddy currents were also calculated from FIDs acquired with nonselective excitation and compared with B0 eddy current values derived using selective excitation. B0 inhomogeneities and center frequency offsets were measured by acquiring FIDs with nonselective excitation. Imaging isocenter shifts were measured using a 33x33x10.5 cm3 uniformity linearity (grid) phantom and a 3D true fast imaging with steady-state precession (TrueFISP) sequence used in MRI-guided radiation therapy. Eddy currents were compared to vendor specifications and correlated with the imaging isocenter shifts. Measurements were conducted before and after the MRI-Linac's waveguide was replaced with an updated design to reduce eddy currents. B0 eddy currents were highly correlated (r = 0.986, P << 0.001) for measurements made with vs without selective excitation. Transverse (X and Y) axis B0 eddy currents before and after the waveguide upgrade were out of specification (specification: ≤0.1 μT m/mT for delays < 10 ms) for most of the measured gantry angles. Gradient eddy currents before and after the upgrade were within specifications for the measured gantry angles (≤0.1% for delays < 10 ms). B0 eddy currents and imaging isocenter shifts were highly correlated (r = 0.965, P << 0.001). After the Linac waveguide upgrade, root mean square (RMS) peak B0 and gradient eddy currents dropped 45% and 11%, respectively, for delays <10 ms, while imaging isocenter shifts dropped 53%. Isocenter shifts were observed in both phase encode and readout directions. Center frequency offsets were <26 Hz while B0 inhomogeneities were <33 Hz full width at half maximum (FWHM). Imaging isocenter shifts measured in a 0.35-T MRI-Linac were highly correlated with B0 eddy currents. The eddy currents and imaging isocenter shifts decreased after the MRI-Linac's waveguide was replaced.


[Brazilian teenagers and beer advertising: relationship between exposure, positive response, and alcohol consumption].
Brazilian teenagers report problematic patterns of alcohol consumption. Alcohol advertising strategies are one of the main factors influencing adolescents' alcohol consumption. The aim of this study was to evaluate the relationship between positive responses to TV beer commercials, exposure, and alcohol consumption. Thirty-two recent TV commercials were shown to 133 high school students from public schools in São Bernardo do Campo, São Paulo State, Brazil. The subjects recorded how well they liked the ads and how often they had already watched each commercial. The teenagers also reported their alcohol consumption rates. The ten commercials analyzed in this article were the five most popular and the five least popular. The analysis showed that subjects had already seen the five most popular ads, but not the five least popular. In addition, the five most popular ads received higher scores from teenagers that reported having consumed beer during the previous month. The study found a positive relationship between enjoying beer advertising and exposure to beer ads, as well as between alcohol consumption and positive responses to alcohol commercials.


The additional benefit of weighted subjective global assessment (SGA) for the predictability of mortality in incident peritoneal dialysis patients: A prospective study.
Although subjective global assessment (SGA) is a widely used tool for nutritional investigation, the scores are dependent on the inspectors' subjective opinions, and there are only few studies that directly assessed the usefulness of SGA and modified SGA in incident peritoneal dialysis (PD) patients. A total of 365 incident PD patients between 2009 and 2015 were enrolled and measured with SGA and calculated using serum albumin and total iron binding capacity (TIBC) levels for weighted SGA. Cox analyses were performed to delineate the association between SGA or weighted SGA and all-cause mortality, and a receiver-operating characteristic was conducted to reveal the additional benefit of weighted SGA on predicting adverse clinical outcomes. The Kaplan-Meier curve showed that the cumulative survival rate in patients with "Good nutrition" (G1) was significantly higher compared to those with "Mild to severe malnutrition" (G2). G2 was significantly associated with an increase in the mortality even after adjusting for several covariates compared with G1. Moreover, a 1-unit increase in weighted SGA was also significantly correlated with mortality after adjustment of the same covariates, while G2 was not significantly associated with an increase in the mortality among young-aged (under 65 years) groups. Meanwhile, a 1-unit increase in weighted SGA was significantly related to an increase in mortality in all the subgroup analyses. Furthermore, the AUCs of weighted SGAs in all groups were significantly increased compared with those of SGA alone. In conclusions, the evaluation of nutritional status based on SGA in incident PD patients might be useful for predicting mortality. However, weighted SGA with serum albumin and TIBC can provide additional predictive power for mortality compared with SGA alone in incident PD patients.


Low-intensity violence and the social determinants of adolescent health among three East African pastoralist communities.
Recently, strong pleas have emerged to place the health of adolescents on the global health agenda. To reposition adolescence front and center, scholars argue that we must work toward a richly contextualized approach that considers the role that social environments play in shaping the final stages of growth and development. We aim to contribute to this deeper understanding of the social determinants of global adolescent health by offering a case study of three nomadic pastoralist communities from northern Kenya. In addition to noteworthy political and economic marginalization, East African pastoralist communities also contend with chronic, low intensity intercommunity conflict. Data collected over five extensive visits from 2008 to 2011, include the 10-19 year olds from 215 randomly sampled Pokot, Samburu, and Turkana households. Using a case/control design, we sampled two sites per ethnic community: one directly affected and one less affected by intercommunity violence. Our nutritional findings indicate that teens ages 15-19 years old had significantly higher anthropometric values compared to younger teens. Living in a wealthier household is associated with greater height, body mass indices, and summed skinfolds for boys but not for girls. Anthropometric measures were influenced by household and community variation in the mixed-effects, multi-level regression models. The Self-Report Questionnaire (SRQ-20) was used to assess psychosocial health, with higher scores associated with living in a community directly affected by violence and having lost a loved one due to violence. Our findings highlight the unique nature of adolescent health challenges but also the central role even subtle differences across communities and households play in shaping young people's experiences. With few studies to document the lived experience of pastoralist youth as they move toward adulthood, examining how such challenging socioeconomic environment shapes health seems long overdue.


Ovarian teratoma associated with anti-N-methyl D-aspartate receptor encephalitis: a report of 5 cases documenting prominent intratumoral lymphoid infiltrates.
Anti-N-methyl D-aspartate receptor (NMDAR) encephalitis is a recently described severe neurological disorder predominantly affecting young women, which presents with psychosis, memory deficits, seizures, and encephalopathy, often requiring prolonged hospitalization. The condition is frequently associated with an underlying neoplasm, most often an ovarian teratoma, and in such cases appears to be a para-neoplastic, immune-mediated encephalopathy. The histologic features of the teratomas associated with anti-NMDAR encephalitis have seldom been described in detail. Therefore, in this report, we have compared ovarian teratomas (4 mature and 1 immature) from 5 patients with anti-NMDAR encephalitis with 22 sporadic control teratomas (14 mature and 8 immature) that included neuroglial elements. The encephalitis-associated tumors ranged from 0.7 to 9.5 cm diameter, and 1 case was bilateral; the second teratoma was discovered 13 mo after the first when symptoms recurred. In comparison with control teratomas, the anti-NMDAR-associated tumors showed a more marked intratumoral lymphoid infiltrate that colocalized to the mature neuroglial elements. Reactive germinal centers (3 cases) and diffuse lymphoplasmacytic infiltrates within the neuroglial matrix (4 cases), and degenerative neuronal changes (2 cases), were seen only in the anti-NMDAR-positive cases. Pathologists encountering ovarian teratomas with these distinctive reactive lymphoid elements should consider the possibility of anti-NMDAR encephalitis, particularly because the neurological symptoms may develop after tumor resection. Careful histopathologic examination may be required to identify small, radiologically occult teratomas, and to demonstrate the presence of subtle neoplastic neuroglial components in teratomas associated with anti-NMDAR encephalitis.


Targeted nanopore sequencing by real-time mapping of raw electrical signal with UNCALLED.
Conventional targeted sequencing methods eliminate many of the benefits of nanopore sequencing, such as the ability to accurately detect structural variants or epigenetic modifications. The ReadUntil method allows nanopore devices to selectively eject reads from pores in real time, which could enable purely computational targeted sequencing. However, this requires rapid identification of on-target reads while most mapping methods require computationally intensive basecalling. We present UNCALLED ( https://github.com/skovaka/UNCALLED ), an open source mapper that rapidly matches streaming of nanopore current signals to a reference sequence. UNCALLED probabilistically considers k-mers that could be represented by the signal and then prunes the candidates based on the reference encoded within a Ferragina-Manzini index. We used UNCALLED to deplete sequencing of known bacterial genomes within a metagenomics community, enriching the remaining species 4.46-fold. UNCALLED also enriched 148 human genes associated with hereditary cancers to 29.6× coverage using one MinION flowcell, enabling accurate detection of single-nucleotide polymorphisms, insertions and deletions, structural variants and methylation in these genes.


Cadmium stress effects indicating marine pollution in different species of sea urchin employed as environmental bioindicators.
In recent years, researches about the defense strategies induced by cadmium stress have greatly increased, invading several fields of scientific research. Mechanisms of cadmium-induced toxicity continue to be of interest for researchers given its ubiquitous nature and environmental distribution, where it often plays the role of pollutant for numerous organisms. The presence in the environment of this heavy metal has been constantly increasing because of its large employment in several industrial and agricultural activities. Cadmium does not have any biological role and, since it cannot be degraded by living organisms, it is irreversibly accumulated into cells, interacting with cellular components and molecular targets. Cadmium is one of the most studied heavy metal inductors of stress and a potent modulator of several processes such as apoptosis, autophagy, reactive oxygen species, protein kinase and phosphatase, mitochondrial function, metallothioneins, and heat-shock proteins. Sea urchins (adults, gametes, embryos, and larvae) offer an optimal opportunity to investigate the possible adaptive response of cells exposed to cadmium, since these cells are known to accumulate contaminants. In this review, we will examine several responses to stress induced by cadmium in different sea urchin species, with a focus on Paracentrotus lividus embryos. The sea urchin embryo represents a suitable system, as it is not subjected to legislation on animal welfare and can be easily used for toxicological studies and as a bioindicator of environmental pollution. Recently, it has been included into the guidelines for the use and interpretation of assays to monitor autophagy.


Disposition of a mixed meal by conscious dogs after seven days of treatment with cyclosporine A and prednisone.
Combination immunosuppressive therapy, that often includes prednisone and cyclosporine A (CyA), is commonly used in the treatment of organ transplant patients. We hypothesized that CyA and prednisone treatment would alter the roles of the liver and peripheral tissues in the disposal of carbohydrates from a meal. Using the arteriovenous difference technique, we examined the disposition of an intragastrically delivered mixed meal in eight 24-hour fasted conscious dogs that had received CyA 15 mg x kg(-1) daily and prednisone 5 mg twice daily for 7 consecutive days before study (CyA-prednisone group). The results were compared with those from a group of 13 dogs (control group) receiving the same meal but no drugs. Neither arterial blood glucose concentrations nor arterial plasma insulin or glucagon concentrations differed significantly between the groups at any time. Cumulative net gut glucose output was equivalent to 43 +/- 9 vs 57% +/- 7% of the glucose in the meal in CyA-prednisone vs control (p = .12). The CyA-prednisone group exhibited greater (p < .05) mean net hepatic glucose uptakes (15.4 +/- 4.6 vs 4.3 +/- 2.2 micromol x kg(-1) x min(-1) and net hepatic fractional extractions of glucose (7.8 +/- 1.6 and 1.5% +/- 1.0%) than the control group. Arterial blood lactate concentrations and net hepatic lactate output were greater in the CyA-prednisone group than the control group (p < .05). Hepatic glycogen content at the end of the study was 2.5-fold greater in the CyA-prednisone group than in the control group (p < .05). The nonhepatic tissues disposed of approximately 91% of the absorbed glucose in the control group but only approximately 26% in the CyA-prednisone group (p < .05). CyA-prednisone treatment caused a marked shift in the carbohydrate disposal from a meal, enhancing the hepatic glucose uptake and decreasing peripheral glucose disposal.


Penetrative anal intercourse may require high axial loading forces.
Despite increases in the lifetime prevalence of anal intercourse, little is known about the safety of inflatable penile prosthesis use during anal intercourse or the forces required for anal penetration. We sought to determine the force required for anal penetration. A digital force gauge device was secured to a silicone phallus with a length and girth representative of the average male phallus in the United States and inserted into the anal canal of 6 male participants for a total of 5 measurements per participant. The maximum axial forces on insertion were recorded. Participants then completed a survey eliciting demographic information and past experience with receptive anal intercourse. The outcome measure was the force recorded during insertion of a force gauge device into the anal canal. The median maximal force recorded on anal penetration was 26.5 N (2.7 kg; range: 16.1-51.7 N or 1.64-5.27 kg). Participants who engaged in receptive anal intercourse more than once per month tended to use lesser median maximal forces (25.7 N or 2.62 kg) than participants who engaged less frequently in receptive anal intercourse (41.6 N or 4.24 kg). Compared with the forces required for vaginal penetration, anal penetration forces may cause additional mechanical stress to the phallus. While the study is limited primarily in its small sample size and in the use of a single device for anal penetration with a set size and rigidity, the resulting findings presented here are to our knowledge the first reported data pertaining the force required for anal penetration. As the forces required for anal penetration exceeded those for vaginal penetration, clinicians may need to carefully counsel patients on the safe use of an inflatable penile prosthesis for anal intercourse.


MEK/ERK signaling directly and indirectly contributes to the cyclical self-renewal of spermatogonial stem cells.
Coordination of stem cell fate is regulated by extrinsic niche signals and stem cell intrinsic factors. In mammalian testes, spermatogonial stem cells maintain constant production of abundant spermatozoa by alternating between self-renewal and differentiation at regular intervals according to a periodical program known as the seminiferous epithelial cycle. Although retinoic acid (RA) signaling has been suggested to direct the cyclical differentiation of spermatogonial stem cells, it remains largely unclear how their cycle-dependent self-renewal/proliferation is regulated. Here, we show that MEK/ERK signaling contributes to the cyclical activity of spermatogonial stem cells. We found that ERK1/2 is periodically activated in Sertoli cells during the stem cell self-renewal/proliferation phase, and that MEK/ERK signaling is required for the stage-related expression of the critical niche factor GDNF. In addition, ERK1/2 is activated in GFRα1-positive spermatogonial stem cells under the control of GDNF and prevent them from being differentiated. These results suggest that MEK/ERK signaling directly and indirectly maintains spermatogonial stem cells by mediating a signal that promotes their periodical self-renewal/proliferation. Conversely, RA signaling directly and indirectly induces differentiation of spermatogonial stem cells. We propose that temporally regulated activations of RA signaling and a signal regulating MEK/ERK antagonistically coordinates the cycle-related activity of spermatogonial stem cells.


Peripheral nerve lesions in hemophilia.
Between 1962 and 1986, eighty-one of the 1351 admissions of patients who had hemophilia to the Nuffield Orthopaedic Centre were for peripheral nerve lesions. Eighty-eight such lesions were identified in fifty-four patients, and thirty-nine of these patients (sixty-one lesions) had adequate follow-up (mean, 8.4 years; range, four months to eighteen years). The femoral nerve was most commonly involved, but involvement of other peripheral nerves also occurred. In thirty (49 per cent) of the sixty-one lesions, the nerve had full motor and sensory recovery; in twenty-one (34 per cent), a residual sensory deficit; and in ten (16 per cent), both a persistent motor and sensory deficit. Patients who had antibodies to factor VIII were significantly less likely to recover full motor or sensory function than were those who did not have such antibodies, and the time to full motor recovery in these patients was significantly longer.


Atomistic to continuum model for studying mechanical properties of RNA nanotubes.
With rapid advancements in the emerging field of RNA nanotechnology, its current and potential applications, new important problems arise in our quest to better understand properties of RNA nanocomplexes. In this paper, our focus is on the modeling of RNA nanotubes which are important for many biological processes. These RNA complexes are also important for human beings, with their theurapeutical and biomedical applications discussed vigorously in the literature over the recent years. Here, we develop a continuum model of RNA nanotubes, originally obtained from self assembly of RNA building blocks in the molecular dynamics simulation. Based on the finite element method, we calculate the elastic properties of these nanostructures and provide a relationship between stress and strain induced in the RNA nanotube. We also analyze the variations in the displacement vector along the assembly axis for RNA nanotubes of different sizes. In particular, we show that oscillations in the amplitudes of strains and displacements significantly differ for such RNA nanotubes. These findings are discussed in the context of atomistic simulations and experimental results in this field.


Epithelial stress and structural remodelling in childhood asthma.
In adult asthma the bronchial epithelium shows high expression of the epidermal growth factor receptor (EGFR) and the cyclin dependent kinase inhibitor, p21waf, linked to ongoing stress and injury. To determine if these are early markers of disease, sections of bronchial specimens obtained post mortem or by bronchoscopy from non-asthmatic (n = 7), moderate (n = 7), or severe (n = 9) asthmatic children aged 5-15 years were examined immunohistochemically. All severe and one moderately asthmatic children were receiving inhaled corticosteroids. The lamina reticularis of the asthmatic biopsy sections was found to be thicker (p = 0.01) than normal with increased deposition of collagen III (p = 0.007); submucosal eosinophil numbers did not differ between groups. As in adults, there was an asthma-related increase in epithelial EGFR (p<0.002) but there was no evidence of proliferation, with Ki67 being reduced (p = 0.001) and p21waf increased (p<0.004). The thickness of the lamina reticularis was significantly correlated with epithelial EGFR (rho = 0.77, p<0.001). These data provide evidence that, in asthmatic children, the epithelium is stressed or injured without significant eosinophilic inflammation. This change in the epithelial phenotype is associated with collagen deposition in the lamina reticularis, suggesting that the epithelial mesenchymal trophic unit is active early in, and may contribute to, the pathogenesis of asthma.


Adenoid cystic carcinoma of the buccal mucosa: a case report with review of literature.
Minor salivary gland neoplasms of the buccal mucosa are relatively uncommon. Adenoid cystic carcinoma (ACC), a well-defined entity, occurs most of the times in the parotid, submandibular glands and palate, as far as the intraoral site is concerned. Adenoid cystic carcinoma tends to have an indolent, extended clinical course with wide local infiltration and late distant metastases. We are presenting a case of an adenoid cystic carcinoma of the buccal mucosa in a 48-year-old female patient.


Sharing Positive Affective States Amongst Rodents.
Group living is thought to benefit from the ability to empathize with others. Much attention has been paid to empathy for the pain of others as an inhibitor of aggression. Empathizing with the positive affect of others has received less attention although it could promote helping by making it vicariously rewarding. Here, we review this latter, nascent literature to show that three components of the ability to empathize with positive emotions are already present in rodents, namely, the ability to perceive, share, and prefer actions that promote positive emotional states of conspecifics. While it has often been argued that empathy evolved as a motivation to care for others, we argue that these tendencies may have selfish benefits that could have stabilized their evolution: approaching others in a positive state can provide information about the source of valuable resources; becoming calmer and optimistic around animals in a calm or positive mood can help adapt to the socially sensed safety level in the environment; and preferring actions also benefiting others can optimize foraging, reduce aggression, and trigger reciprocity. Together, these findings illustrate an emerging field shedding light on the emotional world of rodents and on the biology and evolution of our ability to cooperate in groups.


Dye sensitized solar cells.
Dye sensitized solar cell (DSSC) is the only solar cell that can offer both the flexibility and transparency. Its efficiency is comparable to amorphous silicon solar cells but with a much lower cost. This review not only covers the fundamentals of DSSC but also the related cutting-edge research and its development for industrial applications. Most recent research topics on DSSC, for example, applications of nanostructured TiO(2), ZnO electrodes, ionic liquid electrolytes, carbon nanotubes, graphene and solid state DSSC have all been included and discussed.


Organoaxial partial rotation of duodenum with midgut malrotation in an adult.
Midgut malrotation includes a range of developmental abnormalities that occur during fetal intestinal rotation. Manifestations of intestinal malrotation are generally seen in the paediatric population and are uncommon in adults. Symptomatic patients may present with either acute abdominal pain due to midgut volvulus, or chronic abdominal pain due to proximal midgut partial obstruction in the presence of congenital bands. A limited number of paediatric cases of duodenal occlusion due to duodenal malrotation has been previously reported in the medical literature. We herein report the case of a 57-year-old woman who presented with duodenal obstruction due to organoaxial partial rotation of the distal duodenum associated with midgut malrotation. This is probably the first of such a case diagnosed in adulthood reported in the medical literature. Our patient underwent Roux-en-Y duodenojejunostomy and had symptomatic relief following the successful surgery.


Translocatable resistance to mercuric and phenylmercuric ions in soil bacteria.
Of a sample of 42 gram-negative Hg-resistant bacteria, three (a Pseudomonas fluorescens, a Klebsiella sp. and a Citrobacter sp.) contained translocatable elements conferring resistance to Hg2+ (all three) and to Hg2+ and phenylmercuric acetate (P. fluorescens). The discovery of transposable phenylmercuric acetate resistance extends the range of known resistance "transposons" from heavy metals and antibiotics to organometallic compounds.


[Superior vena cava syndrome caused by dialysis catheter].
The superior vena cava (SVC) syndrome is an uncommon complication due to permanent hemodialysis catheters. Herein we present a case with superior vena cava syndrome resulting from dialysis access catheter placed in the subclavian vein. The patient was admitted with typical signs and symptoms of superior vena cava syndrome. Angiography revealed obstruction of the superior vena cava with thrombosis. Surgical repair consisted of thrombectomy and patch repair of superior vena cava with autologous pericardium. The complaints and symptoms of the patient decreased dramatically following the operation. In view of this case, we do think that creating an early arteriovenous fistula should be kept in mind to minimize the use of dialysis catheter.


Carcass composition and meat quality of equally mature kids and lambs.
Carcass composition and meat quality attributes of 55 suckling kids (27 males and 28 females) and 57 suckling lambs (28 males and 29 females) of Portuguese native breeds were investigated. These suckling kid and lamb meats are European meat quality labels produced according to "Cabrito de Barroso- PGI" and "Borrego Terrincho-PDO" specifications, respectively. Female kids were slaughtered at 9.1 +/- 0.36 kg of BW, and male kids were slaughtered at 10.4 +/- 0.37 kg of BW, corresponding to 20.1 and 17.7% of maturity, respectively. Female lambs were slaughtered at 8.6 +/- 0.53 kg of BW, and male lambs were slaughtered at 9.9 +/- 0.23 kg of BW, corresponding to 19.9 and 17.1% of maturity, respectively. At 24 h postmortem, various yield and quality measurements were collected. The left sides of the carcasses were dissected into muscle, subcutaneous fat, intermuscular fat, and bone. Final pH, instrumental color (L*, a*, b*), carcass measurements, and kidney knob and pelvic fat were also determined. Samples of LM were taken from the lumbar and thoracic cuts for intramuscular and meat quality determinations. At 72 h postmortem, a sample of LM was used for cooking losses and Warner-Bratzler shear force determination. Suckling lambs had greater dressing proportion than suckling kids (P < 0.01). Carcass fatness was not affected by species (P > 0.05), but females had greater kidney knob and pelvic fat proportion than males (P < 0.01). Lambs had greater proportions of the highly valued leg cut and lower proportions of shoulder, anterior rib, and neck cuts than kids. Dissection results indicated that kid carcasses had greater muscle content and lower dissected fat and bone than lambs. Kids had greater (P < 0.001) muscle ultimate pH value than lambs (5.8 +/- 0.02 vs. 5.6 +/- 0.02). Males had greater (P < 0.05) muscle ultimate pH value than females (5.7 +/- 0.02 vs. 5.6 +/- 0.02). The kid meat was significantly lighter (P < 0.05) and less yellow (P < 0.001) than the lamb meat. Kids presented less cooking losses (P < 0.001) than lambs, and shear force value was significantly greater (P < 0.01) in lamb meat. The kid meat had significantly more moisture (P < 0.001) and less intramuscular fat content (P < 0.001) than lambs. At this maturity stage, there were significant differences on both carcass and meat quality attributes of suckling kids and lambs, possibly due to inherent differences between species.


Harnessing synthetic biology for kelp forest conservation1.
Environmental and climatic change is outpacing the ability of organisms to adapt, at an unprecedented level, resulting in range contractions and global ecosystem shifts to novel states. At the same time, scientific advances continue to accelerate, providing never-before imagined solutions to current and emerging environmental problems. Synthetic biology, the creation of novel and engineered genetic variation, is perhaps the fastest developing and transformative scientific field. Its application to solve extant and emerging environmental problems is vast, at times controversial, and technological advances have outpaced the social, ethical, and practical considerations of its use. Here, we discuss the potential direct and indirect applications of synthetic biology to kelp forest conservation. Rather than advocate or oppose its use, we identify where and when it may play a role in halting or reversing global kelp loss and discuss challenges and identify pathways of research needed to bridge the gap between technological advances and organismal biology and ecology. There is a pressing need for prompt collaboration and dialogue among synthetic biologists, ecologists, and conservationists to identify opportunities for use and ensure that extant research directions are set on trajectories to allow these currently disparate fields to converge toward practical environmental solutions.


Bothrops venom-induced hemostasis disorders in the rat: Between Scylla and Charybdis.
Hemostasis impairment represents the most threatening consequence of Viperidae envenoming, notably with Bothrops genus. In the French departments of America, B. atrox envenomation in French Guiana may lead to bleeding while B. lanceolatus envenomation in Martinique to thrombosis. Bleeding related to B. atrox envenomation is attributed to vascular damage mediated by venom metalloproteinases and blood uncoagulable state resulting from thrombocytopenia and consumptive coagulopathy. Thrombosis related to B. lanceolatus envenomation are poorly understood. We aimed to compare the effects of B. atrox and B. lanceolatus venoms in the rat to identify the determinants of the hemorrhagic versus thrombotic complications. Viscoelastometry (ROTEM), platelet count, plasma fibrinogen, thrombin generation assay, fibrinography, endothelial (von Willebrand factor, ADAMTS13 activity, ICAM-1, and soluble E-selectin), and inflammatory biomarkers (IL-1β, IL-6, TNF-α, MCP-1, and PAI-1) were determined in blood samples obtained at H3, H6, and H24 after the subcutaneous venom versus saline injection. In comparison to the control, initial fibrinogen consumption was observed with the two venoms while thrombocytopenia and reduction in the clot amplitude only with B. atrox venom. Moreover, we showed an increase in thrombin generation at H3 with the two venoms, an increase in fibrin generation accompanied with hyperfibrinogenemia at H24 and an increase in inflammatory biomarkers with B. lanceolatus venom. No endothelial damage was found with the two venoms. To conclude, our data support two-sided hemostasis complications in Bothrops envenoming with an initial risk of hemorrhage related to platelet consumption and hypocoagulability followed by an increased risk of thrombosis promoted by the activated inflammatory response and rapid-onset fibrinogen restoration.


Silencing of CD24 Enhances the PRIMA-1-Induced Restoration of Mutant p53 in Prostate Cancer Cells.
In prostate cancer cells, there is CD24-dependent inactivation of mutant p53, but the mechanism and its significance remain largely unknown. Here, we validated this observation and explored the therapeutic potential of targeting CD24 in TP53 mutant prostate cancer cells. Overall, 553 prostate cancers (522 formalin-fixed paraffin-embedded and 31 frozen tissues) were assessed for protein or mRNA expression of CD24 and TP53 The effects of CD24 on p53-dependent transcriptional regulation, cancer cell growth, the cell cycle, apoptosis, and mutant p53 restoration were also determined. As determined with three sample cohorts, CD24 and p53 were not expressed in prostate epithelial cells but in prostate cancer cells in 48% of cases for CD24 and 16% of cases for p53 (mutant form). Expressions of CD24 and mutant p53 were more frequently observed in late-stage and metastatic prostate tumors. Mutant p53 accompanied with CD24 was expressed in most cases (91.6%, 76/83). Silencing of CD24 increased the transcriptional activity of p53 target genes, such as CDKNA1, VDR, and TP53INP1, leading to suppression of p53-dependent cell growth, cell-cycle arrest, and apoptosis in most TP53-mutant prostate cancer cells. Silencing of CD24 enhanced restoration of PRIMA-1-induced mutant p53 in endogenous TP53(P223L/V274F) DU145 cells and in PC3 cells transfected with TP53(R273H) CONCLUSIONS: In human prostate cancers, there is CD24-dependent inactivation of mutant p53. The coexpression of CD24 and p53 may help identify aggressive cancers. Targeting CD24 provides a strategy to enhance mutant p53-restoring therapies, especially in patients with TP53(R273H) prostate cancer. Clin Cancer Res; 22(10); 2545-54. ©2015 AACR.


Selectivity Profiling and Biological Activity of Novel β-Carbolines as Potent and Selective DYRK1 Kinase Inhibitors.
DYRK1A is a pleiotropic protein kinase with diverse functions in cellular regulation, including cell cycle control, neuronal differentiation, and synaptic transmission. Enhanced activity and overexpression of DYRK1A have been linked to altered brain development and function in Down syndrome and neurodegenerative diseases such as Alzheimer's disease. The β-carboline alkaloid harmine is a high affinity inhibitor of DYRK1A but suffers from the drawback of inhibiting monoamine oxidase A (MAO-A) with even higher potency. Here we characterized a series of novel harmine analogs with minimal or absent MAO-A inhibitory activity. We identified several inhibitors with submicromolar potencies for DYRK1A and selectivity for DYRK1A and DYRK1B over the related kinases DYRK2 and HIPK2. An optimized inhibitor, AnnH75, inhibited CLK1, CLK4, and haspin/GSG2 as the only off-targets in a panel of 300 protein kinases. In cellular assays, AnnH75 dose-dependently reduced the phosphorylation of three known DYRK1A substrates (SF3B1, SEPT4, and tau) without negative effects on cell viability. AnnH75 inhibited the cotranslational tyrosine autophosphorylation of DYRK1A and threonine phosphorylation of an exogenous substrate protein with similar potency. In conclusion, we have characterized an optimized β-carboline inhibitor as a highly selective chemical probe that complies with desirable properties of drug-like molecules and is suitable to interrogate the function of DYRK1A in biological studies.


The Frog Skin-Derived Antimicrobial Peptide Esculentin-1a(1-21)NH2 Promotes the Migration of Human HaCaT Keratinocytes in an EGF Receptor-Dependent Manner: A Novel Promoter of Human Skin Wound Healing?
One of the many functions of skin is to protect the organism against a wide range of pathogens. Antimicrobial peptides (AMPs) produced by the skin epithelium provide an effective chemical shield against microbial pathogens. However, whereas antibacterial/antifungal activities of AMPs have been extensively characterized, much less is known regarding their wound healing-modulatory properties. By using an in vitro re-epithelialisation assay employing special cell-culture inserts, we detected that a derivative of the frog-skin AMP esculentin-1a, named esculentin-1a(1-21)NH2, significantly stimulates migration of immortalized human keratinocytes (HaCaT cells) over a wide range of peptide concentrations (0.025-4 μM), and this notably more efficiently than human cathelicidin (LL-37). This activity is preserved in primary human epidermal keratinocytes. By using appropriate inhibitors and an enzyme-linked immunosorbent assay we found that the peptide-induced cell migration involves activation of the epidermal growth factor receptor and STAT3 protein. These results suggest that esculentin-1a(1-21)NH2 now deserves to be tested in standard wound healing assays as a novel candidate promoter of skin re-epithelialisation. The established ability of esculentin-1a(1-21)NH2 to kill microbes without harming mammalian cells, namely its high anti-Pseudomonal activity, makes this AMP a particularly attractive candidate wound healing promoter, especially in the management of chronic, often Pseudomonas-infected, skin ulcers.


HIV drug resistance in HIV positive individuals under antiretroviral treatment in Shandong Province, China.
The efficacy of antiretroviral drugs is limited by the development of drug resistance. Therefore, it is important to examine HIV drug resistance following the nationwide implementation of drug resistance testing in China since 2009. We conducted drug resistance testing in patients who were already on or new to HIV antiretroviral therapy (ART) in Shandong Province, China, from 2011 to 2013, and grouped them based on the presence or absence of drug resistance to determine the effects of age, gender, ethnicity, marital status, educational level, route of transmission and treatment status on drug resistance. We then examined levels of drug resistance the following year. The drug resistance rates of HIV patients on ART in Shandong from 2011 to 2013 were 3.45% (21/608), 3.38% (31/916), and 4.29% (54/1259), per year, respectively. M184V was the most frequently found point mutation, conferring resistance to the nucleoside reverse transcriptase inhibitor, while Y181C, G190A, K103N and V179D/E/F were the most frequent point mutations conferring resistance to the non-nucleoside reverse transcriptase inhibitor. In addition, the protease inhibitor drug resistance mutations I54V and V82A were identified for the first time in Shandong Province. Primary resistance accounts for 20% of the impact factors for drug resistance. Furthermore, it was found that educational level and treatment regimen were high-risk factors for drug resistance in 2011 (P<0.05), while treatment regimen was a high risk factor for drug resistance in 2012 and 2013 (P<0.05). Among the 106 drug-resistant patients, 77 received immediate adjustment of treatment regimen following testing, and 69 (89.6%) showed a reduction in drug resistance the following year. HIV drug resistance has a low prevalence in Shandong Province. However, patients on second line ART regimens and those with low educational level need continuous monitoring. Active drug resistance testing can effectively prevent the development of drug resistance.


α-Amylase immobilization on amidoximated acrylic microfibres activated by cyanuric chloride.
Enzyme immobilization is one of the most important techniques for industrial applications. It makes the immobilized enzyme more stable and advantageous than the free form in different aspects. α-Amylase was immobilized on 4% cyanuric chloride-activated amidoximated acrylic fabric at pH 7.0 with (79%) maximum efficiency. A field emission scanning electron microscope and Fourier transform infrared were used to confirm the immobilization process. Even after being recycled 10 times, the immobilized enzyme lost just 28% of its initial activity. Owing to immobilization, the pH of the soluble α-amylase was shifted from 6.0 to 6.5. The immobilized α-amylases showed thermal stability at 60°C, and became more resistant to heavy metal ions. The k m values of the immobilized and soluble α-amylases were 9.6 and 3.8 mg starch ml-1, respectively. In conclusion, this method shows that the immobilized α-amylase proved to be more efficient than its soluble form, and hence could be used during saccharification of starch.


Cell-mediated haemolytic activity of haemolymph from the Colorado potato beetle (Leptinotarsa decemlineata Say.)
Haemolytic activity was identified in cell-free haemolymph from larval and imago stages of Leptinotarsa decemlineata. The haemolytically active fraction of the haemolymph was active against human, sheep, bull, toad and mouse erythrocytes. There was no haemolysis in the presence of 0.001 M EDTA and 0.5% glutathione. The titre of haemolytic activity did not increase after injury or vaccination of the larvae with Microccocus lysodeikticus. Haemolysin, a heat-labile protein was partially purified by ammonium sulphate precipitation, gel filtration, and ion-exchange separation. SDS PAGE, electrophoresis and immunoblotting showed that the active factor was a protein with a molecular weight of approximately 55 kD. It was not bactericidal for various micro-organisms but the antibacterial activity of the lysozyme increased in the presence of haemolysin only when M. lysodeikticus were used as target cells. Spherulocytes synthesized and released the haemolytic protein in vitro. The haemolytic activity increased in the presence of lipopolysaccharide from Escherichia coli and Ca++ ions. The physiological role of the haemolysin is as yet unknown.


Clostridium septicum infections in children: a case report and review of the literature.
Clostridium myonecrosis is a rare and deadly infection that progresses very rapidly; thus, prompt diagnosis and treatment is vital. In adults, clostridial myonecrosis used to be a well-known complication of war wounds. Today, it is usually seen in settings of trauma, surgery, malignancy, skin infections/burns, and septic abortions. More recently, cases of nontraumatic or spontaneous clostridial myonecrosis have been reported in both adults and children. Clostridium perfringens and Clostridium septicum are responsible for the majority of the clinically relevant infections. Higher mortality rates are seen when C septicum is the causative agent. Here we present a child who survived a severe case of C septicum myonecrosis involving both abdominal and thoracic cavities. This rare infection has a high mortality rate and might be easily misdiagnosed in children, even by experienced clinicians, because of its nonspecific presentation. We also review all reported pediatric cases of C septicum infection and myonecrosis and discuss the surgical and medical interventions associated with improved survival. We identified a total of 47 cases of C septicum infection; of these, 22 (47%) were cases of C septicum associated with myonecrosis. Several factors, if available, were analyzed for each case: age, gender, infection location, previous diagnoses, presenting signs and symptoms, neutropenia, gross pathology of the colon, antibiotic use, surgical intervention, and final outcome. We found that conditions related with C septicum infection in children can be grouped into 3 major categories: patients with neutrophil dysfunction; patients with associated bowel ischemia; and patients with a history of trauma. Malignancies were found in 49% of the cases, cyclic or congenital neutropenia in 21%, hemolytic-uremic syndrome in 11%, structural bowel ischemia in 4%, and local extremity trauma in 6%. In addition, 6% of the cases had no known underlying disorder. Abdominal symptoms including vomiting, diarrhea, blood per rectum, abdominal pain, anorexia, and/or acute abdomen, were reported in 85% of the children. Fever was also a common finding. The mainstay of treatment for C septicum infection was parenteral antibiotics and/or surgical intervention. The mortality rate for children with C septicum infection and myonecrosis was 57% and 59%, respectively. Although 82% of all cases received antibiotics, only 43% underwent therapeutic surgical intervention. Several clinical factors were found to be associated with improved survival. Only 35% of the children with gastrointestinal tract involvement survived, compared with 86% of the children without gastrointestinal tract involvement. The survival rates for other conditions ranged from 0% to 50%. One hundred percent survival was reported in patients with no previously diagnosed conditions and those with infections resulting from trauma to the extremities. All survivors received antibiotic treatment, compared with only 68% of the nonsurvivors. Most survivors (84%) underwent therapeutic surgical intervention, compared with only 12% of nonsurvivors. Other treatments were used adjunctively, including hyperbaric oxygen, granulocyte colony-stimulating factor, granulocyte transfusions, and intravenous immunoglobulin. C septicum infections in children are often fatal; thus, one needs to have a high index of suspicion in at-risk patients. This review describes who these patients are, their clinical presentation, and the therapeutic strategies associated with improved survival.


Bipolar disorder and the endocannabinoid system.
Bipolar disorder (BD) is a debilitating, lifelong neuropsychiatric illness characterised by unsteady mood states which vacillate from (hypo)mania to depression. Despite the availability of pharmaceutical agents which can be effective in ameliorating the acute affective symptoms and prevent episodic relapse, BD is inadequately treated in a subset of patients. The endocannabinoid system (ECS) is known to exert neuromodulatory effects on other neurotransmitter systems critical in governing emotions. Several studies ranging from clinical to molecular, as well as anecdotal evidence, have placed a spotlight on the potential role of the ECS in the pathophysiology of BD. In this perspective, we present advantages and disadvantages of cannabis use in the management of illness course of BD and provide mechanistic insights into how this system might contribute to the pathophysiology of BD. We highlight the putative role of selective cannabinoid receptor 2 (CB2) agonists in BD and briefly discuss findings which provide a rationale for targeting the ECS to assuage the symptoms of BD. Further, data encourage basic and clinical studies to determine how cannabis and cannabinoids (CBs) can affect mood and to investigate emerging CB-based options as probable treatment approaches. The probable role of the ECS has been almost neglected in BD; however, from data available which suggest a role of ECS in mood control, it is justified to support conducting comprehensive studies to determine whether ECS manipulation could positively affect BD. Based on the limited available data, we suggest that activation of CB2 may stabilise mood in this disorder.


Angiogenic activity of human soluble intercellular adhesion molecule-1.
Serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) are elevated in a number of pathological conditions associated with angiogenesis, including tumor growth. Because the increased levels of sICAM-1 suggested that it may be angiogenic, we tested the ability of sICAM-1 to promote angiogenesis. Human recombinant sICAM-1 stimulates chemokinetic endothelial cell migration, endothelial cell tube formation on Matrigel, and sprouting of aortic rings. sICAM-1 also mediates angiogenesis in the chick chorioallantoic membrane assay. Additionally, we found a Mr 49,000 molecule that binds to sICAM-1 that may be the surface ligand on endothelial cells. The evidence that sICAM-1 has angiogenic activity suggests a possible role linking inflammation and neovascularization. Furthermore, sICAM-1 may enhance tumor growth by promoting angiogenesis and escape from immunosurveillance.


The Utility of Real-Time Quantitative Polymerase Chain Reaction Genotype Detection in the Diagnosis of Urinary Tract Infections in Children.
Urinary tract infections (UTIs) are the most common serious bacterial infection in children with significant morbidity with delayed diagnosis. Polymerase chain reaction (PCR) is very accurate in detecting bacteria and widely available, but has never been evaluated to detect UTIs in children. To assess the utility of PCR as a rapid diagnostic tool, we conducted a prospective cohort study of 193 urine samples from children younger than 36 months undergoing evaluation for UTI in the emergency department over a 10-month period. A quantification cycle (Cq) threshold of 26.15 identified all Escherichia coli positive samples with sensitivity and specificity of 100% and 99.5%, respectively (95% CI = 71.5%-100% and 97.9%-99.5%, respectively). A Cq threshold of 19.03 identified E coli infections >100 000 colony forming units/mL with sensitivity and specificity of 100% (95% CI = 72.2%-100% and 98.6%-100%, respectively). PCR is very accurate in diagnosing E coli UTIs in young children and could be useful as a rapid diagnostic tool.


Reversal of Azole Resistance in Candida albicans by Sulfa Antibacterial Drugs.
Invasive candidiasis presents an emerging global public health challenge due to the emergence of resistance to the frontline treatment options, such as fluconazole. Hence, the identification of other compounds capable of pairing with fluconazole and averting azole resistance would potentially prolong the clinical utility of this important group. In an effort to repurpose drugs in the field of antifungal drug discovery, we explored sulfa antibacterial drugs for the purpose of reversing azole resistance in Candida In this study, we assembled and investigated a library of 21 sulfa antibacterial drugs for their ability to restore fluconazole sensitivity in Candida albicans Surprisingly, the majority of assayed sulfa drugs (15 of 21) were found to exhibit synergistic relationships with fluconazole by checkerboard assay with fractional inhibitory concentration index (ΣFIC) values ranging from <0.0312 to 0.25. Remarkably, five sulfa drugs were able to reverse azole resistance in a clinically achievable range. The structure-activity relationships (SARs) of the amino benzene sulfonamide scaffold as antifungal agents were studied. We also identified the possible mechanism of the synergistic interaction of sulfa antibacterial drugs with azole antifungal drugs. Furthermore, the ability of sulfa antibacterial drugs to inhibit Candida biofilm by 40% in vitro was confirmed. In addition, the effects of sulfa-fluconazole combinations on Candida growth kinetics and efflux machinery were explored. Finally, using a Caenorhabditis elegans infection model, we demonstrated that the sulfa-fluconazole combination does possess potent antifungal activity in vivo, reducing Candida in infected worms by ∼50% compared to the control.


PentaLyte decreases lung injury after aortic occlusion-reperfusion.
Lung injury often occurs after hepatoenteric ischemia, with xanthine oxidase (XO, an oxidant-generating enzyme), released from reperfusing liver and intestines, mediating a significant component of this injury. Since pentastarch administration decreases intestinal reperfusion injury, we determined whether resuscitation with PentaLyte (a pentastarch-containing solution) would decrease hepatoenteric reperfusion injury, xanthine oxidase release, and concomitant lung injury after aortic occlusion- reperfusion. Aortic occlusion was established in rabbits for 40 min, and was followed by 3 h of reperfusion, during which either PentaLyte or lactated Ringer's solution-based resuscitation was administered. Sham-operated animals served as controls. Hepatoenteric reperfusion injury, as manifested by release of the enzyme aspartate aminotransferase and decreased gastric intramucosal pH, was significantly (p < 0.0167) attenuated by PentaLyte administration after aortic occlusion-reperfusion, as compared with its occurrence in animals given lactated Ringer's solution. The release of XO after aortic occlusion-reperfusion was 4-fold smaller after PentaLyte administration than after resuscitation with lactated Ringer's solution (p < 0.05). Pulmonary injury, as defined by an increase in bronchoalveolar lavage fluid (BALF) protein content and lactate dehydrogenase (LDH) activity, was 4-fold less after PentaLyte administration following aortic occlusion-reperfusion than after administration of lactated Ringer's solution (p < 0.05). We conclude that remote pulmonary injury is significantly decreased by concomitant PentaLyte-mediated reduction of hepatoenteric reperfusion injury and XO release.


Anomaly in aortic arch alters pathological outcome of transient global ischemia in Rhesus macaques.
We investigated a non-human primate (NHP) transient global ischemia (TGI) model which was induced by clipping the arteries originating from the aortic arch. Previously we demonstrated that our TGI model in adult Rhesus macaques (Macaca mulatta) results in marked neuronal cell loss in the hippocampal region, specifically the cornu Ammonis (CA1) region. However, we observed varying degrees of hippocampal cell loss among animals. Here, we report for the first time an anomaly of the aortic arch in some Rhesus macaques that appears as a key surgical factor in ensuring the success of the TGI model in this particular NHP. Eleven adult Rhesus macaques underwent the TGI surgery, which involved 10-15-minute clipping of both innominate and subclavian arteries. Animals were allowed to survive between 1 day and 28 days after TGI. Because of our experience and knowledge that Japanese macaques exhibited only innominate and subclavian arteries arising from the aortic arch, macroscopic visualization of these two arteries alone in the Rhesus macaques initially assured us that clipping both arteries was sufficient to produce TGI. During the course of one TGI operation, however, we detected 3 arterial branches arising from the aortic arch, which prompted us to subsequently search for 3 branches in succeeding TGI surgeries. In addition, we performed post-mortem examination of the heart to confirm the number of arterial branches in the aortic arch. Finally, in order to reveal the pathological effect of the aortic arch anomaly, we compared the hippocampal cell loss between animals found to have 3 arterial branches but had all or only two branches clipped during TGI operation. Post-mortem examination revealed that eight NHPs had the typical two arterial aortic branches, but three NHPs displayed an extra arterial aortic branch, indicating that about 30% of Rhesus macaques had 3 arterial branches arising from the aorta. Histological analyses using Nissl staining showed that in NHPs with the aortic arch anomaly clipping only two of three arterial branches led to a partial cell loss and minimal alteration in number of cell layers in the hippocampal region when compared with clipping all three branches, with the hippocampal cell death in the latter resembling the pathological outcome achieved by clipping the two arterial branches in NHPs displaying the typical two-artery aortic arch. The finding that 3 of 11 NHPs exhibited an extra arterial aortic branch recognizes this aortic arch anomaly in Rhesus macaques that warrants a critical surgical maneuver in order to successfully produce consistent TGI-induced hippocampal cell loss.


Can resistance training impact MRI outcomes in relapsing-remitting multiple sclerosis?
Multiple sclerosis (MS) is characterised by accelerated brain atrophy, which relates to disease progression. Previous research shows that progressive resistance training (PRT) can counteract brain atrophy in other populations. To evaluate the effects of PRT by magnetic resonance imaging (MRI) and clinical measures of disease progression in people with MS. This study was a 24-week randomised controlled cross-over trial, including a Training ( n = 18, 24 weeks of PRT followed by self-guided physical activity) and Waitlist group ( n = 17, 24 weeks of habitual lifestyle followed by PRT). Assessments included disability measures and MRI (lesion load, global brain volume, percentage brain volume change (PBVC) and cortical thickness). While the MS Functional Composite score improved, Expanded Disability Status Scale, lesion load and global brain volumes did not differ between groups. PBVC tended to differ between groups and higher absolute cortical thickness values were observed in 19 of 74 investigated cortical regions after PRT. Observed changes were confirmed and reproduced when comparing relative cortical thickness changes between groups for four areas: anterior cingulate gyrus, temporal pole, orbital sulcus and inferior temporal sulcus. PRT seem to induce an increase in cortical thickness, indicating that PRT have a neuroprotective or even neuroregenerative effect in relapsing-remitting MS.


Consumption of crustaceans by megaherbivorous dinosaurs: dietary flexibility and dinosaur life history strategies.
Large plant-eating dinosaurs are usually presumed to have been strictly herbivorous, because their derived teeth and jaws were capable of processing fibrous plant foods. This inferred feeding behavior offers a generalized view of dinosaur food habits, but rare direct fossil evidence of diet provides more nuanced insights into feeding behavior. Here we describe fossilized feces (coprolites) that demonstrate recurring consumption of crustaceans and rotted wood by large Late Cretaceous dinosaurs. These multi-liter coprolites from the Kaiparowits Formation are primarily composed of comminuted conifer wood tissues that were fungally degraded before ingestion. Thick fragments of laminar crustacean cuticle are scattered within the coprolite contents and suggest that the dinosaurian defecators consumed sizeable crustaceans that sheltered in rotting logs. The diet of decayed wood and crustaceans offered a substantial supply of plant polysaccharides, with added dividends of animal protein and calcium. Nevertheless, it is unlikely that the fossilized fecal residues depict year-round feeding habits. It is more reasonable to infer that these coprolites reflected seasonal dietary shifts-possibly related to the dinosaurs' oviparous breeding activities. This surprising fossil evidence challenges conventional notions of herbivorous dinosaur diets and reveals a degree of dietary flexibility that is consistent with that of extant herbivorous birds.


Effect of early treatment with propranolol on left ventricular function four weeks after myocardial infarction.
Left ventricular function and exercise capacity were assessed in 79 patients randomised to receive intravenous and oral propranolol (n = 44) or conventional therapy (n = 35) within four hours of onset of their first myocardial infarction. Cineangiocardiography and exercise testing were performed four weeks after infarction to allow for maximum recovery of myocardial function. Left ventriculography showed no improvement in ejection fraction or preservation of regional contractile function in patients treated with propranolol compared with controls. A trend towards smaller end diastolic volumes was seen in the propranolol group (mean (SD) 151(42) ml) compared with controls (167(42) ml). Exercise duration and frequency of angina were not significantly different in the two groups. It is concluded that limitation of infarct size by propranolol does not lead to a significant improvement in ventricular systolic function, although left ventricular dilatation may be reduced. These findings are consistent with the known effect of early intravenous beta blockade which limits infarct size by preservation of subepicardial myocardium.


In vivo evaluation of limiting brain penetration of probes for α(2C)-adrenoceptor using small-animal positron emission tomography.
To evaluate in vivo brain penetration of α(2C)-adrenoceptor (α(2C)-AR) antagonists as a therapeutic agent, we synthesized two new (11)C-labeled selective α(2C)-AR antagonists 4-(6,7-dimethoxy-1,2,3,4-tetrahydroisoquinolin-2-yl)methyl-2-aryl-7-methoxybenzofuran ([(11)C]MBF) and acridin-9-yl-[4-(4-methylpiperazin-1-yl)phenyl]amine ([(11)C]JP-1302) as α(2C)-AR-selective positron emission tomography (PET) probes. The radiochemical yield, specific activity, and radiochemical purity of these probes was appropriate for injection. To evaluate whether the brain penetration of these probes is related to the function of two major drug efflux transporters, P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP), we performed PET studies using wild-type and P-gp/Bcrp knockout mice. In wild-type mice, the radioactivity level after injection with [(11)C]MBF initially increased and effluxed immediately from the brain, whereas that with [(11)C]JP-1302 was distributed throughout the brain. However, the regional distribution of radioactivity after injection with [(11)C]JP-1302 in the brain was different from that of α(2C)-ARs. In P-gp/Bcrp knockout mice, uptake of [(11)C]MBF was approximately 3.7-fold higher and that of [(11)C]JP-1302 was approximately 1.6-fold higher than those in wild-type mice. These results indicate that brain penetration of the two PET probes was affected by modulation of P-gp and Bcrp functions.


Biomedical challenges of human senescence: a review.
To summarise and discuss the progress made in the study of human senescence over the past one hundred years and assess the achievements to date. Published original research and reviews during the past one hundred years. The summary focused on those contributions that tested the various hypotheses that attempt to identify and explain the factors that are involved in the ageing process. Online and manual library searches provided a body of data on which the summaries and discussions were based. Specific questions were addressed: Why does ageing occur? What are the key mechanisms? To what extent are genetic and environmental factors involved in the ageing process? How does the immune system behave during ageing and especially against infectious agents? Answers to these questions were discussed against the background of major improvements in life expectancy in most parts of the world except for sub-Saharan Africa where the HIV/AIDS pandemic has reversed the trend. Biological and clinical studies over the past century clearly reflect a better understanding of the major factors involved in human senescence. It is appreciated that human life expectancy has improved dramatically over the period through achievements in public health, therapy, nutrition and general living standards. A great deal remains to be done through multidisciplinary research before the quality of life can be improved further.


Role of the precuneus in the detection of incongruency between tactile and visual texture information: A functional MRI study.
Visual clues as to the physical substance of manufactured objects can be misleading. For example, a plastic ring can appear to be made of gold. However, we can avoid misidentifying an object׳s substance by comparing visual and tactile information. As compared to the spatial properties of an object (e.g., orientation), however, little information regarding physical object properties (material properties) is shared between vision and touch. How can such different kinds of information be compared in the brain? One possibility is that the visuo-tactile comparison of material information is mediated by associations that are previously learned between the two modalities. Previous studies suggest that a cortical network involving the medial temporal lobe and precuneus plays a critical role in the retrieval of information from long-term memory. Here, we used functional magnetic resonance imaging (fMRI) to test whether these brain regions are involved in the visuo-tactile comparison of material properties. The stimuli consisted of surfaces in which an oriented plastic bar was placed on a background texture. Twenty-two healthy participants determined whether the orientations of visually- and tactually-presented bar stimuli were congruent in the orientation conditions, and whether visually- and tactually-presented background textures were congruent in the texture conditions. The texture conditions revealed greater activation of the fusiform gyrus, medial temporal lobe and lateral prefrontal cortex compared with the orientation conditions. In the texture conditions, the precuneus showed greater response to incongruent stimuli than to congruent stimuli. This incongruency effect was greater for the texture conditions than for the orientation conditions. These results suggest that the precuneus is involved in detecting incongruency between tactile and visual texture information in concert with the medial temporal lobe, which is tightly linked with long-term memory.


[Biochemical mechanisms of the antihypoxic effect of amichlorphen (diethylaminoethylamide parachlorphenoxyacetic acid)].
Test set up on albino rats demonstrated a preliminary introduction of amichlophene to prevent a hypoxic fall in the level of highly ergastic compounds in the tissues owing to stimulation of the mitochondrial respiration processes and increased energy-producting effectiveness of the latter. The beneficial effect of amichlophen on the bioenergetics of the tissues in acute hypoxia is largely caused by the continued structural and functional integrity of the mitochondria, lysosomes and other cellular organellae which is due to the inhibitory action of the compound on the prosses of lipids reoxidation and to the activity of phospholipase A.


Open ear hearing aids in tinnitus therapy: An efficacy comparison with sound generators.
This study aimed to compare the effectiveness of tinnitus retraining therapy (TRT) with sound generators or with open ear hearing aids in the rehabilitation of tinnitus for a group of subjects who, according to Jastreboff categories, can be treated with both approaches to sound therapy (borderline of Category 1 and 2). This study was a prospective data collection with a parallel-group design which entailed that each subject was randomly assigned to one of the two treatments group: half of the subjects were fitted binaurally with sound generators, and the other half with open ear hearing aids. Both groups received the same educational counselling sessions. Ninety-one subjects passed the screening criteria and were enrolled into the study. Structured interviews, with a variety of measures evaluated through the use of visual-analog scales and the tinnitus handicap inventory self-administered questionnaire, were performed before the therapy and at 3, 6, and 12 months during the therapy. Data showed a highly significant improvement in both tinnitus treatments starting from the first three months and up to one year of therapy, with a progressive and statistically significant decrease in the disability every three months. TRT was equally effective with sound generator or open ear hearing aids: they gave basically identical, statistically indistinguishable results.


Annulus upsizing for mitral valve re-replacement in children.
Mitral valve replacement remains the last resort for treatment of congenital mitral valve disease. Enlarging the mitral annulus at the time of mitral valve replacement may allow implantation of a larger prosthesis in children. All mitral valve replacement procedures from 1990 to 2012 were included, and mitral annulus enlargement techniques were analyzed. A control group of patients undergoing replacement of a previously implanted mechanical mitral valve prosthesis was compared with the cohort of patients who underwent annular enlarging procedures. Primary end points were increases in annulus size and Z score, and freedom from paced rhythm. Among 205 mitral valve replacement procedures, 16 (8%) included techniques to upsize the mitral annulus in 15 patients, all but 1 of whom had undergone prior mitral valve replacement. These patients were compared with a control cohort of 53 patients undergoing mitral valve re-replacement without annulus enlargement. The annulus was upsized by 18% ± 11% (compared with 16% ± 20% in controls, P = .5) using open balloon dilation in 4 patients, radial annular incisions in 5 patients, and patch augmentation of the aorto-mitral continuity in 7 patients. The mean valvar diameter Z score increased from -0.2 ± 1.3 to +1.1 ± 1.4 (P < .001), compared with an increase from -0.3 ± 1.2 to 0.9 ± 1.1 in controls (P < .001). During a mean follow-up of 6.5 ± 6.4 years, 4 of 12 patients required permanent pacing in the upsizing group (predominantly with patch augmentation), versus 5 of 53 patients in the control group. In children undergoing mitral valve replacement, various techniques can be used to upsize the mitral annulus and allow implantation of a larger prosthesis. There is a nontrivial risk of heart block with annulus upsizing, which deserves further study.


Slow membrane potential changes in skeletal muscle induced in Cl(-)-free medium.
1. Conventional microelectrode techniques were used to measure simultaneous changes in membrane potential (Vm) and conductance (Gm) induced by single electrical stimuli in muscles bathed in Cl(-)-free solution containing 40 mM of tetraethylammonium (TEA+). 2. Stimulation induced slow transient depolarizations (slow response) accompanied by increased calcium conductance, while the potassium conductance was first elevated and later reduced. 3. Stepwise elevation of [K+]0 from 2.5 to 5 or 10 mM during the slow response evoked an abrupt repolarization of 42.3 +/- 8.9 mV (n = 4; p less than 0.001), and 24.8 +/- 3.5 +/- mV (n = 5; p less than 0.001), respectively, while Gm was increased to 1.45 +/- 0.25-fold (n = 5; p less than 0.05). Neither the slow response nor K(+)-induced changes in Vm or Gm were sensitive to tetrodotoxin (3 microM), however, nifedipine (10 microM) abolised the slow response. 4. It was concluded that beyond the increase of calcium conductance, the ionic conductance of the inward rectifier K+ channel was reduced during the slow response, which could be restored by the elevation of [K+]0. The results suggest the possible contribution of these mechanisms to the electrical instability of myotonic muscles. Potential therapeutic consequences are discussed.


Absorb Bioresorbable Scaffold Versus Xience Metallic Stent for Prevention of Restenosis Following Percutaneous Coronary Intervention in Patients at High Risk of Restenosis: Rationale and Design of the COMPARE ABSORB Trial.
The advent of bioresorbable vascular scaffolds (BVS) was considered as a potential improvement in percutaneous coronary intervention (PCI) after the groundbreaking development of drug eluting stents (DES). However, the clinical performance, long-term safety and efficacy of BVS in complex coronary lesions remain uncertain. COMPARE ABSORB, a multicenter, single blind, prospective randomized trial, aims to compare the clinical outcomes between the Absorb BVS and Xience everolimus-eluting metallic stent (EES) in patients with coronary artery disease and a high risk of restenosis. COMPARE ABSORB is designed to enroll 2100 patients at up to 45 European sites. Enrolled patients will possess high risk for restenosis due to clinical profile or coronary lesion complexity and will undergo elective or emergent PCI. Once included in the study, patients will receive either Absorb BVS or Xience EES. Specific advice on implantation technique including mandatory pre-dilatation, sizing and post-dilatation (PSP), will be used in the Absorb BVS arm. The primary endpoint is target lesion failure (TLF), a device-oriented composite endpoint (cardiac death, target vessel myocardial infarction and clinically-indicated target lesion revascularization). The trial is powered to assess non-inferiority of Absorb BVS compared with Xience EES with a predetermined non-inferiority margin of 4.5% at 1 year after index procedure. The clinical follow-up will continue for 7 years. The prospective COMPARE ABSORB randomized trial (ClinicalTrials.govNCT02486068) will help to assess the long-term safety and efficacy of Absorb BVS compared with Xience EES in the treatments of patients with complex coronary artery disease and a high attendant risk of restenosis.


GPR55 regulates cannabinoid 2 receptor-mediated responses in human neutrophils.
The directional migration of neutrophils towards inflammatory mediators, such as chemokines and cannabinoids, occurs via the activation of seven transmembrane G protein coupled receptors (7TM/GPCRs) and is a highly organized process. A crucial role for controlling neutrophil migration has been ascribed to the cannabinoid CB(2) receptor (CB(2)R), but additional modulatory sites distinct from CB(2)R have recently been suggested to impact CB(2)R-mediated effector functions in neutrophils. Here, we provide evidence that the recently de-orphanized 7TM/GPCR GPR55 potently modulates CB(2)R-mediated responses. We show that GPR55 is expressed in human blood neutrophils and its activation augments the migratory response towards the CB(2)R agonist 2-arachidonoylglycerol (2-AG), while inhibiting neutrophil degranulation and reactive oxygen species (ROS) production. Using HEK293 and HL60 cell lines, along with primary neutrophils, we show that GPR55 and CB(2)R interfere with each other's signaling pathways at the level of small GTPases, such as Rac2 and Cdc42. This ultimately leads to cellular polarization and efficient migration as well as abrogation of degranulation and ROS formation in neutrophils. Therefore, GPR55 limits the tissue-injuring inflammatory responses mediated by CB(2)R, while it synergizes with CB(2)R in recruiting neutrophils to sites of inflammation.