Predictors of incomplete aneurysm occlusion after treatment with the Pipeline Embolization Device: PREMIER trial 1 year analysis.
Flow diverters have revolutionized the treatment of intracranial aneurysms. Nevertheless, some aneurysms fail to occlude with flow diversion. The Prospective Study on Embolization of Intracranial Aneurysms with the Pipeline Device (PREMIER) was a prospective, multicenter and single-arm trial of small and medium wide-necked unruptured aneurysms. In the current study, we evaluate the predictors of treatment failure in the PREMIER cohort. We analyzed PREMIER patients who had incomplete occlusion (Raymond-Roy >1) at 1 year angiographic follow-up and compared them with those who achieved Raymond-Roy 1, aiming to identify predictors of treatment failure. 25 aneurysms demonstrated incomplete occlusion at 1 year. There was a median reduction of 0.9 mm (IQR 0.41-2.43) in maximum diameter between pre-procedure and 1 year measurements, with no aneurysmal hemorrhage. Patients with incomplete occlusion were significantly older than those with complete occlusion (p=0.011). Smoking (p=0.045) and C6 segment location (p=0.005) were significantly associated with complete occlusion, while location at V4 (p=0.01) and C7 (p=0.007) and involvement of a side branch (p<0.001) were significantly associated with incomplete occlusion. In multivariable logistic regression, significant predictors of incomplete occlusion were non-smoker status (adjusted OR 4.49, 95% CI 1.11 to 18.09; p=0.03) and side branch involvement (adjusted OR 11.68, 95% CI 3.84 to 35.50; p<0.0001), while C6 location had reduced odds of incomplete occlusion (adjusted OR 0.29, 95% CI 0.10 to 0.84; p=0.02). The results of our study are consistent with previous retrospective series and warrant consideration for technique adaptations to achieve higher occlusion rates. Further follow-up is needed to assess progression of aneurysm occlusion and clinical behavior in these cases.


Enrichment of Phosphatidylethanolamine in Viral Replication Compartments via Co-opting the Endosomal Rab5 Small GTPase by a Positive-Strand RNA Virus.
Positive-strand RNA viruses build extensive membranous replication compartments to support replication and protect the virus from antiviral responses by the host. These viruses require host factors and various lipids to form viral replication complexes (VRCs). The VRCs built by Tomato bushy stunt virus (TBSV) are enriched with phosphatidylethanolamine (PE) through a previously unknown pathway. To unravel the mechanism of PE enrichment within the TBSV replication compartment, in this paper, the authors demonstrate that TBSV co-opts the guanosine triphosphate (GTP)-bound active form of the endosomal Rab5 small GTPase via direct interaction with the viral replication protein. Deletion of Rab5 orthologs in a yeast model host or expression of dominant negative mutants of plant Rab5 greatly decreases TBSV replication and prevents the redistribution of PE to the sites of viral replication. We also show that enrichment of PE in the viral replication compartment is assisted by actin filaments. Interestingly, the closely related Carnation Italian ringspot virus, which replicates on the boundary membrane of mitochondria, uses a similar strategy to the peroxisomal TBSV to hijack the Rab5-positive endosomes into the viral replication compartments. Altogether, usurping the GTP-Rab5-positive endosomes allows TBSV to build a PE-enriched viral replication compartment, which is needed to support peak-level replication. Thus, the Rab family of small GTPases includes critical host factors assisting VRC assembly and genesis of the viral replication compartment.


Hypothalamic hamartoma: epilepsy and neurodevelopmental profiles in a clinical cohort
We aimed to determine the prevalence of epilepsy and neurodevelopmental disorders, including autism spectrum disorder, in children and adolescents with hypothalamic hamartoma (HH). We also sought to explore the relationship between these neurodevelopmental comorbidities and epilepsy and to establish the predictive value of structural characteristics of the hamartoma itself. We retrospectively studied a cohort of 62 children with HH, with neuroimaging reviewed at Great Ormond Street Hospital (GOSH) between 2008 and 2018. Clinical records were reviewed, cognitive and language data analysed, and MRI scans studied. We confirmed a high burden of epilepsy (56%), autism (19%) and other neurodevelopmental disorders. Although rates of some neurodevelopmental disorders were significantly higher in those with epilepsy, autistic features and/or early developmental concerns often predated the onset of seizures, in particular generalized seizures, or occurred independently of seizures. We found a significant correlation between certain structural characteristics of the hamartoma itself and both epilepsy and certain neurodevelopmental comorbidities. These findings suggest that although seizure burden clearly contributes to the cognitive and behavioural phenotypes seen, the hamartoma itself, and particular characteristics of it, are likely to be primary determinants of both the epilepsy and neurodevelopmental profiles. It is also probable that the underlying aetiology, likely genetic, directly contributes to the clinical profile, with epilepsy, neurodevelopmental impairment and the hamartoma itself representing markers of this aetiology. We propose that atypical neurodevelopmental profiles in HH could best be conceptualized as a developmental and epileptic encephalopathy. These findings have implications for counselling, monitoring and treatment.


A note on interactions between temperature, viscosity, body size and swimming energetics in fish larvae
In a previous study, it was shown that at a given speed the larvae of a species of freshwater fish, the Danube bleak Chalcalburnus chalcoides, expended considerably more metabolic energy at 15 degreesC than at 20 degreesC. We applied hydromechanical arguments to our previous data in order to determine whether the higher cost of swimming at the lower temperature might be due to the effects of viscous forces. However, even under the unrealistic assumption of the larvae swimming in the viscous regime at Reynolds numbers as high as 2000, we show here that hydromechanical forces cannot explain the high energy cost of swimming at 15 degreesC. Instead, we offer a new hypothesis that the 'two-gear system' of the swimming muscles operating in juvenile and adult fish is not yet functional in the larvae, with the consequence that, when these fish are swimming at high speeds in cold water, the muscle fibres have to operate over an increasingly inefficient range of shortening velocities.


Evaluation of the efficiency and effectiveness of independent dose calculation followed by machine log file analysis against conventional measurement based IMRT QA.
Experimental methods are commonly used for patient-specific IMRT delivery verification. There are a variety of IMRT QA techniques which have been proposed and clinically used with a common understanding that not one single method can detect all possible errors. The aim of this work was to compare the efficiency and effectiveness of independent dose calculation followed by machine log file analysis to conventional measurement-based methods in detecting errors in IMRT delivery. Sixteen IMRT treatment plans (5 head-and-neck, 3 rectum, 3 breast, and 5 prostate plans) created with a commercial treatment planning system (TPS) were recalculated on a QA phantom. All treatment plans underwent ion chamber (IC) and 2D diode array measurements. The same set of plans was also recomputed with another commercial treatment planning system and the two sets of calculations were compared. The deviations between dosimetric measurements and independent dose calculation were evaluated. The comparisons included evaluations of DVHs and point doses calculated by the two TPS systems. Machine log files were captured during pretreatment composite point dose measurements and analyzed to verify data transfer and performance of the delivery machine. Average deviation between IC measurements and point dose calculations with the two TPSs for head-and-neck plans were 1.2 ± 1.3% and 1.4 ± 1.6%, respectively. For 2D diode array measurements, the mean gamma value with 3% dose difference and 3 mm distance-to-agreement was within 1.5% for 13 of 16 plans. The mean 3D dose differences calculated from two TPSs were within 3% for head-and-neck cases and within 2% for other plans. The machine log file analysis showed that the gantry angle, jaw position, collimator angle, and MUs were consistent as planned, and maximal MLC position error was less than 0.5 mm. The independent dose calculation followed by the machine log analysis takes an average 47 ± 6 minutes, while the experimental approach (using IC and 2D diode array measurements) takes an average about 2 hours in our clinic. Independent dose calculation followed by machine log file analysis can be a reliable tool to verify IMRT treatments. Additionally, independent dose calculations have the potential to identify several problems (heterogeneity calculations, data corruptions, system failures) with the primary TPS, which generally are not identifiable with a measurement-based approach. Additionally, machine log file analysis can identify many problems (gantry, collimator, jaw setting) which also may not be detected with a measurement-based approach. Machine log file analysis could also detect performance problems for individual MLC leaves which could be masked in the analysis of a measured fluence.


[Prevalence and risk factors for diabetic retinopathy in type 2 diabetes patients in Jewish and Bedouin populations in southern Israel].
Diabetic retinopathy is the most common microvascular complication of diabetes. In recent years, there is a dramatic increase in the number of diabetic patients in the Bedouin population in the Negev region. To analyze the clinical features and find out the incidence and risk factors of diabetic retinopathy of type 2 diabetes patients in the Jewish and Bedouin populations. Data was collected from the files of 523 patients, who were examined by ophthalmologists at different clinics in southern Israel, and who were not previously diagnosed with diabetic retinopathy until that examination. All the data was analyzed by univariate analysis, and a multivariate model was built to predict the risk to develop diabetic retinopathy, separately for the Jewish and Bedouin population in the Negev. The average age was 64 +/- 10.3 years in the Jewish population and 58.6 +/- 12 years in the Bedouin population (P < 0.001). Treatment with insulin was administered to 14.5% of the Bedouin population, compared to 5.7% of the Jewish population (P < 0.001). Eye complications (diabetic retinopathy and/or maculopathy) were found in 13.4% of Jews, compared to 22% of Bedouins (P < 0.01). In the multivariate analysis we found 6 predicting factors for the development of diabetic retinopathy in the Jewish population: long duration of diabetes, older age, high HbA1c, insulin treatment, high levels of LDL and creatinine; and 4 predicting factors in the Bedouin population: long duration of diabetes, high HbA1c, insulin treatment and smoking. The Bedouin population in southern Israel suffers more from retinal diabetic complications compared to Jewish patients. Common risk factors for both populations are long duration of diabetes, high HbA1c and insulin treatment.


[Psychopathology of person misidentification--a multidimensional concept].
The misidentification of person or Personenverkennung (PV) is provisionally redefined in this paper by focusing on four important points of view offered by German-speaking authors, all more or less based on an analysis of content. These deal with the extent of "craziness" of a PV (Leonhard), the degree of personal relationship of the person misidentified with respect to the patient (Kahlbaum/Pauleikhoff), the extent of the incorrect identification (Jaspers), and, lastly, the stage of stability shown by a PV at a particular point in time (Conrad). On this basis it is suggested that it may be best to initially call the PV simply a "delusion" with all further differentiation of types of PV carried out in light of a multidimensional psychopathological model utilizing various combinations of these four aspects of content.


Effect of ramipril on walking times and quality of life among patients with peripheral artery disease and intermittent claudication: a randomized controlled trial.
Approximately one-third of patients with peripheral artery disease experience intermittent claudication, with consequent loss of quality of life. To determine the efficacy of ramipril for improving walking ability, patient-perceived walking performance, and quality of life in patients with claudication. Randomized, double-blind, placebo-controlled trial conducted among 212 patients with peripheral artery disease (mean age, 65.5 [SD, 6.2] years), initiated in May 2008 and completed in August 2011 and conducted at 3 hospitals in Australia. Patients were randomized to receive 10 mg/d of ramipril (n = 106) or matching placebo (n = 106) for 24 weeks. Maximum and pain-free walking times were recorded during a standard treadmill test. The Walking Impairment Questionnaire (WIQ) and Short-Form 36 Health Survey (SF-36) were used to assess walking ability and quality of life, respectively. At 6 months, relative to placebo, ramipril was associated with a 75-second (95% CI, 60-89 seconds) increase in mean pain-free walking time (P < .001) and a 255-second (95% CI, 215-295 seconds) increase in maximum walking time (P < .001). Relative to placebo, ramipril improved the WIQ median distance score by 13.8 (Hodges-Lehmann 95% CI, 12.2-15.5), speed score by 13.3 (95% CI, 11.9-15.2), and stair climbing score by 25.2 (95% CI, 25.1-29.4) (P < .001 for all). The overall SF-36 median Physical Component Summary score improved by 8.2 (Hodges-Lehmann 95% CI, 3.6-11.4; P = .02) in the ramipril group relative to placebo. Ramipril did not affect the overall SF-36 median Mental Component Summary score. Among patients with intermittent claudication, 24-week treatment with ramipril resulted in significant increases in pain-free and maximum treadmill walking times compared with placebo. This was associated with a significant increase in the physical functioning component of the SF-36 score. clinicaltrials.gov Identifier: NCT00681226.


Cardioprotection by H2S engages a cGMP-dependent protein kinase G/phospholamban pathway.
H2S is known to confer cardioprotection; however, the pathways mediating its effects in vivo remain incompletely understood. The purpose of the present study is to evaluate the contribution of cGMP-regulated pathways in the infarct-limiting effect of H2S in vivo. Anaesthetized rabbits were subjected to myocardial ischaemia (I)/reperfusion (R), and infarct size was determined in control or H2S-exposed groups. The H2S donor sodium hydrosulfide (NaHS, an agent that generates H2S) increased cardiac cGMP and reduced the infarct size. The cGMP-dependent protein kinase (PKG)-I inhibitor DT2 abrogated the protective effect of NaHS, whereas the control peptide TAT or l-nitroarginine methyl ester (l-NAME) did not alter the effect of NaHS. Moreover, the KATP channel inhibitor, glibenclamide, partially reversed the effects of NaHS, whereas inhibition of mitochondrial KATP did not modify the NaHS response. NaHS enhanced phosphorylation of phospholamban (PLN), in a PKG-dependent manner. To further investigate the role of PLN in H2S-mediated cardioprotection, wild-type and PLN KO mice underwent I/R. NaHS did not exert cardioprotection in PLN KO mice. Unlike what was observed in rabbits, genetic or pharmacological inhibition of eNOS abolished the infarct-limiting effect of NaHS in mice. Our findings demonstrate (i) that administration of NaHS induces cardioprotection via a cGMP/PKG/PLN pathway and (ii) contribution of nitric oxide to the H2S response is species-specific.


Solution-focused counselling in Canadian outpatient dietetic practice.
The use of solution-focused counselling (SFC) in dietetics was examined to determine the demand for an SFC teaching tool for registered dietitians. A cross-sectional web-based survey was conducted within the Dietitians of Canada cardiology, eating disorders, and consulting networks, with dietitians who practised in an outpatient setting. Dietitians were surveyed on the seven core components of SFC. Responses (n=58) were received from dietitians practising in seven provinces, who tended to be younger (mean age=44) and who had practised for a mean of 15 years. The entire sample reported use of at least four of the seven core components of SFC. All respondents were interested in receiving a counselling teaching tool for dietitians. All respondents were using components of SFC in their outpatient practices. The efficaciousness of SFC in dietetic practice has yet to be determined.


Investigation into the animal species contents of popular wet pet foods.
The use of the generic term "meat and animal derivatives" in declared ingredient lists of pet foods in the European Union is virtually universal. In the wake of the 2013 "horse meat scandal" in the human food chain, we examined the presence and authenticity of animal sources (cow, chicken, pig and horse) of proteins in a range of popular wet pet foods in the United Kingdom. Seventeen leading dog and cat foods were sampled for the relative presence of DNA from each of the four animal species by quantitative real-time polymerase chain reaction. No horse DNA was detected. However, there was detection at substantial levels of unspecified animal species in most products tested. In 14 out of 17 samples, bovine, porcine and chicken DNA were found in various proportions and combinations but were not explicitly identified on the product labels. Of the 7 products with prominent headline descriptions containing the term "with beef", only 2 were found to contain more bovine DNA (>50%) than pig and chicken DNA combined. There is a need for the pet food industry to show greater transparency to customers in the disclosure of the types of animal proteins (animal species and tissue types) in their products. Full disclosure of animal contents will (a) allow more informed choices to be made on purchases which are particularly important for pets with food allergies, (b) reduce the risk of product misinterpretation by shoppers, and


Ovarian cancer G protein-coupled receptor 1 deficiency exacerbates crystal deposition and kidney injury in oxalate nephropathy in female mice.
Ovarian cancer G protein-coupled receptor 1 (OGR1) (Gpr68) and G protein-coupled receptor 4 (GPR4) (Gpr4) are proton-activated G protein-coupled receptors that are stimulated upon increased extracellular acidity. These receptors have various physiological and pathophysiological roles in renal acid-base physiology, tissue inflammation, and fibrosis among others. Their function in injured renal tissue, however, remains mostly unclear. To address this, we investigated their role in crystalline nephropathy by increasing the oxalate intake of GPR4 KO and OGR1 KO mice. After 10 days of high-oxalate intake and 4 days of recovery, renal crystal content, histopathology, filtration function, and inflammation were assessed. While GPR4 deficiency did not show major alterations in disease progression, OGR1 KO mice had higher urinary calcium levels and exacerbated crystal accumulation accompanied by decreased creatinine clearance and urea excretion and a decreased presence of regulatory T (Treg) cells in kidney tissue. When lowering the severity of the kidney injury, OGR1 KO mice were more prone to develop crystalline nephropathy. In this setting, OGR1 KO mice displayed an increased activation of the immune system and a higher production of proinflammatory cytokines by T cells and macrophages. Taken together, in the acute setting of oxalate-induced nephropathy, the lack of the proton-activated G protein-coupled receptor (GPCR) GPR4 does not influence disease. OGR1 deficiency, however, increases crystal deposition leading to impaired kidney function. Thus, OGR1 may be important to limit kidney crystal deposition, which might subsequently be relevant for the pathophysiology of oxalate kidney stones or other crystallopathies.


Standardized cryopreservation of human primary cells.
Cryopreservation is the use of low temperatures to preserve structurally intact living cells. The cells that survive the thermodynamic journey from the 37 °C incubator to the -196 °C liquid nitrogen storage tank are free from the influences of time. Thus, cryopreservation is a critical component of cell culture and cell manufacturing protocols. Successful cryopreservation of human cells requires that the cells be derived from patient samples that are collected in a standardized manner, and carefully handled from blood draw through cell isolation. Furthermore, proper equipment must be in place to ensure consistency, reproducibility, and sterility. In addition, the correct choice and amount of cryoprotectant agent must be added at the correct temperature, and a controlled rate of freezing (most commonly 1 °C/min) must be applied prior to a standardized method of cryogenic storage. This appendix describes how human primary cells can be frozen for long-term storage and thawed for growth in a tissue culture vessel.


Structural basis for the bypass of the major oxaliplatin-DNA adducts by human DNA polymerase η.
Oxaliplatin, together with cisplatin, is among the most important drugs used in cancer chemotherapy. Oxaliplatin, which contains a bulky diaminocyclohexane (DACH) moiety, kills cancer cells mainly by producing (DACH)Pt-GpG intrastrand cross-links that impede transcription. The Pt-GpG tolerance by translesion DNA synthesis (TLS) polymerases contributes to the resistance of tumors to platinum-based chemotherapy. In particular, human DNA polymerase η (Polη) readily bypasses Pt-GpG adducts. While many structural studies have addressed how TLS polymerases interact with cisplatin-DNA adducts, a structure of DNA polymerase in complex with oxaliplatin-DNA adducts has not been reported, limiting our understanding of bypass of the bulky (DACH)Pt-GpG lesion by TLS polymerases. Herein, we report the first structure of DNA polymerase bound to oxaliplatinated DNA. We determined a crystal structure of Polη incorporating dCTP opposite the 3'G of the (DACH)Pt-GpG, which provides insights into accurate, efficient bypass of the oxaliplatin-GpG adducts by TLS polymerases. In the catalytic site of Polη, the 3'G of the (DACH)Pt-GpG formed three Watson-Crick hydrogen bonds with incoming dCTP and the primer terminus 3'-OH was optimally positioned for nucleotidyl transfer. To accommodate the bulky (DACH)Pt-GpG lesion, the Val59-Trp64 loop in the finger domain of Polη shifted from the positions observed in the corresponding Polη-cisplatin-GpG and undamaged structures, suggesting that the flexibility of the Val59-Trp64 loop allows the enzyme's bypass of the (DACH)Pt-GpG adducts. Overall, the Polη-oxaliplatin-GpG structure provides a structural basis for TLS-mediated bypass of the major oxaliplatin-DNA adducts and insights into resistance to platinum-based chemotherapy in humans.


Acclimatory responses of Arabidopsis to fluctuating light environment: comparison of different sunfleck regimes and accessions.
Acclimation to fluctuating light environment with short (lasting 20 s, at 650 or 1,250 μmol photons m(-2) s(-1), every 6 or 12 min) or long (for 40 min at 650 μmol photons m(-2) s(-1), once a day at midday) sunflecks was studied in Arabidopsis thaliana. The sunfleck treatments were applied in the background daytime light intensity of 50 μmol photons m(-2) s(-1). In order to distinguish the effects of sunflecks from those of increased daily irradiance, constant light treatments at 85 and 120 μmol photons m(-2) s(-1), which gave the same photosynthetically active radiation (PAR) per day as the different sunfleck treatments, were also included in the experiments. The increased daily total PAR in the two higher constant light treatments enhanced photosystem II electron transport and starch accumulation in mature leaves and promoted expansion of young leaves in Columbia-0 plants during the 7-day treatments. Compared to the plants remaining under 50 μmol photons m(-2) s(-1), application of long sunflecks caused upregulation of electron transport without affecting carbon gain in the form of starch accumulation and leaf growth or the capacity of non-photochemical quenching (NPQ). Mature leaves showed marked enhancement of the NPQ capacity under the conditions with short sunflecks, which preceded recovery and upregulation of electron transport, demonstrating the initial priority of photoprotection. The distinct acclimatory responses to constant PAR, long sunflecks, and different combinations of short sunflecks are consistent with acclimatory adjustment of the processes in photoprotection and carbon gain, depending on the duration, frequency, and intensity of light fluctuations. While the responses of leaf expansion to short sunflecks differed among the seven Arabidopsis accessions examined, all plants showed NPQ upregulation, suggesting limited ability of this species to utilize short sunflecks. The increase in the NPQ capacity was accompanied by reduced chlorophyll contents, higher levels of the xanthophyll-cycle pigments, faster light-induced de-epoxidation of violaxanthin to zeaxanthin and antheraxanthin, increased amounts of PsbS protein, as well as enhanced activity of superoxide dismutase. These acclimatory mechanisms, involving reorganization of pigment-protein complexes and upregulation of other photoprotective reactions, are probably essential for Arabidopsis plants to cope with photo-oxidative stress induced by short sunflecks without suffering from severe photoinhibition and lipid peroxidation.


Work flow policy and within-worker and between-workers variability in performance.
Work flow policies are shown to induce a change in average between-workers variability (worker heterogeneity) and within-worker variability in performance times. In a laboratory experiment, the authors measured the levels of worker heterogeneity and within-worker variability under an individual performance condition, a work sharing condition, and a fixed assignment condition. The work sharing policy increased the levels of worker heterogeneity and worker variability, whereas the fixed assignment policy decreased them. These effects, along with work flow policy main effects on mean performance times and variability are examined. This article represents an initial step in understanding effects that may be important in the selection of an operating policy, the ignorance of which may lead to costly misestimates of performance.


Diosgenin ameliorates testicular damage in streptozotocin-diabetic rats through attenuation of apoptosis, oxidative stress, and inflammation.
Diabetes mellitus (DM) is a prevalent metabolic disorder that is associated with development of some complications in male reproductive system including testicular damage, sexual dysfunction, abnormal spermatogenesis, and infertility. Diosgenin is a natural steroidal saponin with anti-diabetic, anti-oxidative, and anti-inflammatory effects. This research study was undertaken to explore the protective effect of diosgenin against diabetes-induced testicular damage in the rat. Ten days following streptozotocin (STZ; i.p.), diosgenin was daily administered for 6 weeks (p.o.). Diosgenin administration to diabetic rats significantly improved body weight and lowered serum glucose. In addition, diosgenin-treated diabetic group had a significantly lower level of malondialdehyde (MDA), protein carbonyl, greater level of glutathione (GSH), and higher activity of superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx) in addition to testicular improvement of ferric reducing antioxidant power (FRAP). Furthermore, diosgenin significantly improved serum insulin and testosterone level and alleviated testicular markers of inflammation including tumor necrosis factor α (TNFα) and interleukin 6 (IL-6) in diabetic rats. Moreover, apoptotic markers including caspase 3 activity, Annexin V, and DNA fragmentation decreased, mitochondrial membrane potential (MMP) accentuated, and myeloperoxidase (MPO) activity as a biomarker of neutrophil infiltration decreased in diosgenin-treated diabetic group. Additionally, diosgenin was capable to improve sperm count, motility, and viability in addition to prevention of damage to seminiferous tubules in diabetic animals. Collectively, diosgenin ameliorates testicular damage in DM, at least via partial suppression of apoptosis, oxidative stress, inflammation, and neutrophil infiltration and also via partial restoration of mitochondrial integrity.


Pyrazine analogs are active components of wolf urine that induce avoidance and fear-related behaviors in deer.
Our previous studies indicated that a cocktail of pyrazine analogs, identified in wolf urine, induced avoidance and fear behaviors in mice. The effects of the pyrazine cocktail on Hokkaido deer (Cervus nippon yesoensis) were investigated in field bioassays at a deer park in Hokkaido, Japan. A set of feeding bioassay trials tested the effects of the pyrazine cocktail odor on the behavior of the deer located around a feeding area in August and September 2013. This odor effectively suppressed the approach of the deer to the feeding area. In addition, the pyrazine cocktail odor provoked fear-related behaviors, such as "tail-flag", "flight" and "jump" actions, of the deer around the feeding area. This study is the first experimental demonstration that the pyrazine analogs in wolf urine have robust and continual fearful aversive effects on ungulates as well as mice. The pyrazine cocktail might be suitable for a chemical repellent that could limit damage to forests and agricultural crops by wild ungulates.


Lower risk of atopic disorders in whole cell pertussis-vaccinated children.
This study addressed whether whole cell pertussis-vaccinated children have a different risk of atopic disorders compared with children who did not receive this vaccination. Data on vaccination status, atopic disorders and child and family characteristics of the children of 700 families were collected in this retrospective study. A minority of these 700 families refused vaccinations for religious reasons. The relation between pertussis-vaccination status and atopic disorders was analysed by means of adjusted logistic regression for repeated measurements in order to account for the correlation between sibship members. The 700 families included 1,961 children. Data on vaccination status and atopic disorders were available for 1,724 children. Vaccinated children had a reduced risk of atopic disorders. Whole cell pertussis vaccination is associated with a lower risk of atopic disorders, though other vaccine components (diphtheria, tetanus, poliomyelitis) or other vaccinations may also be involved.


Exploring the psychosis functional connectome: aberrant intrinsic networks in schizophrenia and bipolar disorder.
Intrinsic functional brain networks (INs) are regions showing temporal coherence with one another. These INs are present in the context of a task (as opposed to an undirected task such as rest), albeit modulated to a degree both spatially and temporally. Prominent networks include the default mode, attentional fronto-parietal, executive control, bilateral temporal lobe, and motor networks. The characterization of INs has recently gained considerable momentum, however; most previous studies evaluate only a small subset of the INs (e.g., default mode). In this paper we use independent component analysis to study INs decomposed from functional magnetic resonance imaging data collected in a large group of schizophrenia patients, healthy controls, and individuals with bipolar disorder, while performing an auditory oddball task. Schizophrenia and bipolar disorder share significant overlap in clinical symptoms, brain characteristics, and risk genes which motivates our goal of identifying whether functional imaging data can differentiate the two disorders. We tested for group differences in properties of all identified INs including spatial maps, spectra, and functional network connectivity. A small set of default mode, temporal lobe, and frontal networks with default mode regions appearing to play a key role in all comparisons. Bipolar subjects showed more prominent changes in ventromedial and prefrontal default mode regions whereas schizophrenia patients showed changes in posterior default mode regions. Anti-correlations between left parietal areas and dorsolateral prefrontal cortical areas were different in bipolar and schizophrenia patients and amplitude was significantly different from healthy controls in both patient groups. Patients exhibited similar frequency behavior across multiple networks with decreased low frequency power. In summary, a comprehensive analysis of INs reveals a key role for the default mode in both schizophrenia and bipolar disorder.


Metabolism of glucose 1,6-P2. I. Enzymes involved in the synthesis of glucose 1,6-P2 in pig tissues.
Pig tissues show four enzymatic activities of glucose 1,6-P2 synthesis: (A) 2 [glucose 1-P]----glucose 1,6-P2 + glucose; (B) glucose 1-P + ATP----glucose 1,6-P2 + ADP; (C) glucose 1-P + fructose 1,6-P2----glucose 1,6-P2 + fructose 6-P; (D) glucose 1-P + glycerate 1,3-P2----glucose 1,6-P2 + glycerate 3-P. Brain is the tissue with highest capability of glucose 1,6-P2 synthesis. With the exception of skeletal muscle, activity "D" represents the highest activity of glucose 1,6-P2 synthesis. In muscle, activity "B" is the major activity. The existence of a specific glucose 1,6-P2 synthase which catalyzes reaction "D" is confirmed. Two peaks of such an enzyme are isolated by ion-exchange chromatography. There is an enzyme which specifically catalyzes reaction "C", not previously described. There is a glucose 1-P kinase not identical to phosphofructokinase.


[Transrectal recatheterization under echocardiography control].
We present the case of a 36 years-old woman, in whom antegrade mitral intraluminal valvulotomy was performed under transesophageal echocardiography control. Inadvertently, we retired the Inoue's catheter to the right atrium from the left atrium, before the dilation of the mitral valve was accomplished. Under echocardiographic control we reintroduced the transseptal catheter across the former septal orifice, avoiding a new septal puncture, and its possible complications. Now a days, the antegrade intraluminal mitral valvulotomy is easy performed, because the transesophageal echocardiography monitoring. The transesophageal echocardiography has modified the transseptal catheterization contraindications.


On the perception of time.
In this article, we review scientific work and present new results on the perception of time, that is, on the feeling of time as perceived by individuals. The phenomenon of time being felt passing faster with growing age is well known, and there are numerous interesting studies to shed light on the question why this is so. Many of these are based on studies in psychology and social sciences. Others range from symptoms of the ageing process to related symptoms of decreasing memory capacities. Again other explanations, quite different in nature from the preceding ones, involve event intensities in the life of individuals. The relative decrease of interesting new events as one grows older is seen as an important factor contributing to the feeling that time is thinned out. The last type of possible explanations can be made more explicit in a mathematical model. Quantitative conclusions about the rate of decrease of the feeling of time can be drawn, and, interestingly, without restrictive assumptions. It is shown that under this model the feeling of time is thinned out at least logarithmically. Numerical constants will depend on specific hypotheses which we discuss but the lower-bound logarithmic character of the thinning-out phenomenon does not depend much on these. The presented model can be generalized in several ways. In particular we prove that there are, a priori, no logical incompatibilities in a model leading to the very same distribution of time perception for individuals with completely different pace and style of life. Our model is built to explain long-time perception. No claim is made that the feeling of time being thinned out is omnipresent for very individual. However, this is typically the case and we explain why.


Unusual meningeal tuberculoma--case report.
A 60-year-old male presented with bilateral papilledema. Neuroimaging showed a densely enhanced flat tumor within the interhemispheric region in the frontal and parietal lobes with sulcal obliteration and enhancement. There was no history of tuberculous infection. Histological examination of the excised tissue from the flat tumor within the interhemispheric region was consistent with tuberculoma. Multiple antituberculous agent therapy resolved the papilledema and reduced the tumor.


Mechanism of Alkyne Hydroarylation Catalyzed by (P,C)-Cyclometalated Au(III) Complexes.
Over the last 5-10 years, gold(III) catalysis has developed rapidly. It often shows complementary if not unique features compared to gold(I) catalysis. While recent work has enabled major synthetic progress in terms of scope and efficiency, very little is yet known about the mechanism of Au(III)-catalyzed transformations and the relevant key intermediates have rarely been authenticated. Here, we report a detailed experimental/computational mechanistic study of the recently reported intermolecular hydroarylation of alkynes catalyzed by (P,C)-cyclometalated Au(III) complexes. The cationic (P,C)Au(OAcF)+ complex (OAcF = OCOCF3) was authenticated by mass spectrometry (MS) in the gas phase and multi-nuclear NMR spectroscopy in solution at low temperatures. According to density functional theory (DFT) calculations, the OAcF moiety is κ2-coordinated to gold in the ground state, but the corresponding κ1-forms featuring a vacant coordination site sit only slightly higher in energy. Side-on coordination of the alkyne to Au(III) then promotes nucleophilic addition of the arene. The energy profiles for the reaction between trimethoxybenzene (TMB) and diphenylacetylene (DPA) were computed by DFT. The activation barrier is significantly lower for the outer-sphere pathway than for the alternative inner-sphere mechanism involving C-H activation of the arene followed by migratory insertion. The π-complex of DPA was characterized by MS. An unprecedented σ-arene Au(III) complex with TMB was also authenticated both in the gas phase and in solution. The cationic complexes [(P,C)Au(OAcF)]+ and [(P,C)Au(OAcF)(σ-TMB)]+ stand as active species and off-cycle resting state during catalysis, respectively. This study provides a rational basis for the further development of Au(III) catalysis based on π-activation.


Silica xerogels as pharmaceutical drug carriers.
This review focuses on silica xerogels obtained by the sol-gel method and their application as drug delivery systems. SiO(2) xerogels are potential biomaterials to be used as matrix materials for the extended and controlled release of different kinds of biologically active agents administered by various routes. The article includes some representative examples that describe the encapsulation of bioactive molecules and model compounds inside a silica matrix produced by the conventional sol-gel method or by ultrasound hydrolysis. The drug release rate from xerogels could be modified by adjusting several parameters, such as the type of precursor, the concentration of the catalyst and drying temperature. In vitro and in vivo studies have shown the efficacy and biodegradability of these composites. The potential application of silica xerogels as drug carrier systems is critically analyzed and discussed.


Water exchange of collagenous tissues and of gelatin.
Water exchange of parenchymatous tissue, namely liver, kidney, or pancreas, occurs by osmosis, and the movement of water, as is well known, occurs in direct relation to the concentration of the surrounding solution. But the present work shows that the hydration of collagenous tissues, like that of gelatin, occurs in strong as well as in weak solutions of sodium chloride. The water intake of collagenous tissue in solutions of sodium chloride or of sucrose increases with increased density of the tissue and the sequence of changes is like that observed with gels of increasing gelatin content under the same conditions. Dense collagenous tissue, apparently impervious to the movement of water, exhibits a conspicuous ability to attract and hold it.


Neurophysiological Predictors of Response to Medication in Parkinson's Disease.
Background: Although dopaminergic medication has been the foundation of Parkinson's disease (PD) therapy for decades, sensitive and specific therapeutic response biomarkers that allow for better treatment optimization are lacking. Objective: We tested whether the features of Transcranial Magnetic Stimulation-based neurophysiological measures taken off-medication are associated with dopaminergic medication-induced clinical effects. Method: Motor cortex excitability [short-latency intracortical inhibition (SICI), intracortical facilitation (ICF), short-latency afferent inhibition (SAI), and input-output (IO) curve], and plasticity [paired associative stimulation (PAS) protocol] neurophysiological measures were examined in 23 PD patients off-medication. Clinical features were quantified by the motor section of the Unified Parkinson's Disease Scale (total score and lateralized total, bradykinesia, and rigidity sub-scores), and the differences between measures off-medication and on-medication (following the usual morning dose), were determined. Total daily dopaminergic medication dose (expressed as levodopa equivalent daily dose-LEDD), was also determined. Results: SICI significantly correlated with changes in lateralized UPDRS motor and bradykinesia sub-scores, suggesting that patients with stronger basal intracortical inhibition benefit more from dopaminergic treatment than patients with weaker intracortical inhibition. Also, ICF significantly negatively correlated with LEDD, suggesting that patients with stronger intracortical facilitation require less dopaminergic medication to achieve optimal therapeutic benefit. Both associations were independent of disease severity and duration. Conclusions: The results suggest variability of (patho) physiological phenotypes related to intracortical inhibitory and facilitatory mechanisms determining clinical response to dopaminergic medication in PD. Measures of intracortical excitability may help predict patients' response to dopaminergic therapy, thus potentially providing a background for developing personalized therapy in PD.


Microelectrodes in medicine.
The chief applications of microelectrodes in medicine are in experimental studies of tissue and cell behaviour and of metabolism. Microprobes have been developed for a wide range of specific substrates and for the detection of bioelectric potentials. They have provided information that now forms the basis of much of our understanding of biological phenomena. Their inherent characteristics, such as fragility and 'drift', render them unsuitable for general clinical application except at the surface of organs. The heterogeneity of tissues makes it necessary to specify the exact anatomical location of microelectrode measurements and the probes themselves must be of a form that neither distorts, damages, nor stimulates reactions from, the cells or blood vessels with which they are in contact.


Influence of Bushen Tiaochong Cycle Therapy on Th1/Th2 Deviation, Sex Hormone Level, and Pregnancy Outcome of Alloimmune Recurrent Spontaneous Abortion.
To analyze the influence of Bushen Tiaochong cycle therapy on Th1/Th2 deviation, sex hormone level, and pregnancy outcome of alloimmune recurrent spontaneous abortion (RSA). From August 2018 to April 2020, 130 patients with alloimmune RSA who met the inclusion criteria were randomly divided into the control group (n = 65) and the study group (n = 65). The former received lymphocyte immunotherapy (LIT), and the latter received Bushen Tiaochong cycle therapy on the basis of LIT. The treatment ended at 12 w of pregnancy. The total score of traditional Chinese medicine (TCM) syndromes, Th1 cytokine (IL-2), Th2 cytokine (IL-10), and related hormones (chorionic gonadotropin (hCG) and progesterone (P)) were compared between the two groups before and after treatment. The positive rate of blocking antibody (BA), pregnancy success rate, and preterm birth rate were counted. After treatment, the total score of TCM syndromes, IL-2 level, and Th1/Th2 ratio in the two groups decreased significantly, while the levels of IL-10, hCG, and P increased significantly, and the study group improved significantly compared with the control group (P < 0.05). The positive rate of BA and pregnancy success rate in the study group were higher than those in the control group (P < 0.05). There was no significant difference in the preterm birth rate between the two groups (P > 0.05). On the basis of routine western medicine treatment, a combined application of Bushen Tiaochong cycle therapy can significantly improve the Th1/Th2 deviation, serum sex hormone level, and pregnancy outcome in patients with alloimmune RSA.


Increased turnover of dopamine in caudate nucleus of detoxified alcoholic patients.
A previous study of the DOPA decarboxylase substrate 6-[(18)F]fluoro-L-DOPA (FDOPA) with positron emission tomography (PET) detected no difference of the net blood-brain transfer rate (Kin(app)) between detoxified alcoholic patients and healthy controls. Instead, the study revealed an inverse correlation between Kin (app) in left ventral striatum and alcohol craving scores. To resolve the influx and efflux phases of radiolabeled molecules, we independently estimated the unidirectional blood-brain FDOPA clearance rate (K) and the washout rate of [(18)F]fluorodopamine and its deaminated metabolites (k(loss)), and we also calculated the total distribution volume of decarboxylated metabolites and unmetabolized FDOPA as a steady-state index of the dopamine storage capacity (V(d)) in brain. The craving scores in the 12 alcoholics correlated positively with the rate of loss (k(loss)) in the left ventral striatum. We conclude that craving is most pronounced in the individuals with relatively rapid dopamine turnover in the left ventral striatum. The blood-brain clearance rate (K), corrected for subsequent loss of radiolabeled molecules from brain, was completely normal throughout the brain of the alcoholics, in whom the volume of distribution (V(d)) was found to be significantly lower in the left caudate nucleus. The magnitude of Vd in the left caudate head was reduced by 43% relative to the 16 controls, consistent with a 58% increase of k(loss). We interpret the findings as indicating that a trait for rapid dopamine turnover in the ventral striatum subserves craving and reward-dependence, leading to an acquired state of increased dopamine turnover in the dorsal striatum of detoxified alcoholic patients.


Epigallocatechin-3-gallate inhibits collagen production of nasal polyp-derived fibroblasts.
Nasal polyps are chronic inflammatory conditions characterized by myofibroblast differentiation and extracelluar matrix accumulation. The major catechin from green tea is (-)-epigallocatechin-3-gallate (EGCG), which has garnered attention for its potential to prevent oxidative stress-related diseases. The purpose of this study was twofold: (i) to determine the effect of EGCG on fibroblast differentiation into myofibroblasts and extracellular matrix accumulation in transforming growth factor (TGF)-β1-induced nasal polyp-derived fibroblasts (NPDFs) and (ii) to determine if the antioxidative effect of EGCG on reactive oxygen species (ROS) production in TGF-β1-induced NPDFs is involved in the aforementioned processes. TGF-β1-induced NPDFs were treated with or without EGCG. α-smooth muscle actin (α-SMA) and collagen type I mRNA were analyzed by reverse transcription-polymerase chain reaction. α-SMA protein was also detected using immunofluorescent staining. The amount of total soluble collagen was analyzed by Sircol collagen assay. ROS activity was measured by the nitroblue tetrazolium reduction assay and visualized by fluorescent microscopy. EGCG significantly inhibited expressions of α-SMA and collagen type I mRNA and reduced α-SMA and collagen protein levels at concentrations of 10-20 µg/mL. EGCG also inhibited TGF-β1-induced ROS production at the same concentrations. These results suggest the possibility that EGCG may be effective at inhibiting the development of nasal polyps through an anti-oxidant effect.


Efficient Mass Spectral Analysis of Active Transporters Overexpressed in Escherichia coli.
Structural analysis of purified active membrane proteins can be performed by mass spectrometry (MS). However, no large-scale expression systems for active eukaryotic membrane proteins are available. Moreover, because membrane proteins cannot easily be digested by trypsin and ionized, they are difficult to analyze by MS. We developed a method for mass spectral analysis of eukaryotic membrane proteins combined with an overexpression system in Escherichia coli. Vesicular glutamate transporter 2 (VGLUT2/SLC17A6) with a soluble α-helical protein and histidine tag on the N- and C-terminus, respectively, was overexpressed in E. coli, solubilized with detergent, and purified by Ni-NTA affinity chromatography. Proteoliposomes containing VGLUT2 retained glutamate transport activity. For MS analysis, the detergent was removed from purified VGLUT2 by trichloroacetic acid precipitation, and VGLUT2 was then subjected to reductive alkylation and tryptic digestion. The resulting peptides were detected with 88% coverage by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS with or without liquid chromatography. Vesicular excitatory amino acid transporter and vesicular acetylcholine transporter were also detected with similar coverage by the same method. Thus this methodology could be used to analyze purified eukaryotic active transporters. Structural analysis with chemical modifiers by MS could have applications in functional binding analysis for drug discovery.


Traumatic Optic Nerve Sheath Hematoma.
The aim of this report is to present a patient with traumatic optic nerve sheath hematoma (ONSH), a rare diagnosis with high potential for visual sequelae. This case involves a 41-year-old male who presented promptly following blunt trauma to the right eye and orbit that resulted in acute vision loss. Following computed tomography and ophthalmic examination, a diagnosis of ONSH was made and medical therapy with methylprednisolone was initiated. He reported significant improvements in visual symptoms following intravenous corticosteroid therapy. Although the patient reported significant improvements and had normal Snellen visual acuities in follow-up, he continued to have an inferior visual field defect at 1 week in the affected eye. ONSH causing subsequent localized compression of the optic nerve is a rare mechanism of traumatic optic neuropathy in patients following head trauma. The localized compartment syndrome of the optic nerve and subjective visual symptoms were relieved following corticosteroid therapy with no initial need for surgical decompression. Although central visual acuity returned to baseline, the patient had a persistent visual field defect and relative afferent pupillary defect.


Tracking of pitch probabilities in congenital amusia.
Auditory perception involves not only hearing a series of sounds but also making predictions about future ones. For typical listeners, these predictions are formed on the basis of long-term schematic knowledge, gained over a lifetime of exposure to the auditory environment. Individuals with a developmental disorder known as congenital amusia show marked difficulties with music perception and production. The current study investigated whether these difficulties can be explained, either by a failure to internalise the statistical regularities present in music, or by a failure to consciously access this information. Two versions of a melodic priming paradigm were used to probe participants' abilities to form melodic pitch expectations, in an implicit and an explicit manner. In the implicit version (Experiment 1), participants made speeded, forced-choice discriminations concerning the timbre of a cued target note. In the explicit version (Experiment 2), participants used a 1-7 rating scale to indicate the degree to which the pitch of the cued target note was expected or unexpected. Target notes were chosen to have high or low probability in the context of the melody, based on the predictions of a computational model of melodic expectation. Analysis of the data from the implicit task revealed a melodic priming effect in both amusic and control participants whereby both groups showed faster responses to high probability than low probability notes rendered in the same timbre as the context. However, analysis of the data from the explicit task revealed that amusic participants were significantly worse than controls at using explicit ratings to differentiate between high and low probability events in a melodic context. Taken together, findings from the current study make an important contribution in demonstrating that amusic individuals track melodic pitch probabilities at an implicit level despite an impairment, relative to controls, when required to make explicit judgments in this regard. However the unexpected finding that amusics nevertheless are able to use explicit ratings to distinguish between high and low probability notes (albeit not as well as controls) makes a similarly important contribution in revealing a sensitivity to musical structure that has not previously been demonstrated in these individuals.


Monoglyceride-based organogelator for broad-range oil uptake with high capacity.
Oil/water separation has been a worldwide subject because of increasing release of oil-containing wastewater as well as several marine oil spills. The phase-selective organogelators (PSOGs) are thought to offer a potential and effective implement for addressing this issue. An ideal PSOG for oil adsorption must fulfill some requirements involving effective gelation, easy synthesis, low cost, and recyclable for reuse. However, beyond those, the ability of gelation for a broad-range oil phase without selectivity is also important. However, most of the reported PSOGs have limitation in this respect thus far. In this paper, a new class of saturated 1-monoglyceride-derived organogelators can efficiently uptake almost all of the common fuel oils from water and gelate organic solvents with extremely low minimum gelation concentration (MGC). In addition, the oils in the gel and gelator molecules can be recovered quantitatively through simple vacuum distillation.


Impact of smoke-free housing policy lease exemptions on compliance, enforcement and smoking behavior: A qualitative study.
This paper investigates the impacts of smoke-free housing policies on compliance, enforcement and smoking behavior. From 2012 to 2014, we studied two affordable housing providers in Canada with comprehensive smoke-free policies: Waterloo Regional Housing that required new leases to be non-smoking and exempted existing leases, and Yukon Housing Corporation that required all leases (existing and new) to be non-smoking. Focus groups and key informant interviews were conducted with 31 housing and public health staff involved in policy development and implementation, and qualitative interviews with 56 tenants. Both types of smoke-free policies helped tenants to reduce and quit smoking. However, exempting existing tenants from the policy created challenges for monitoring compliance and enforcing the policy, and resulted in ongoing tobacco smoke exposure. Moreover, some new tenants were smoking in exempted units, which undermined the policy and maintained smoking behavior. Our findings support the implementation of complete smoke-free housing policies that do not exempt existing leases to avoid many of the problems experienced by staff and tenants. In jurisdictions where exempting existing leases is still required by law, adequate staff resources for monitoring and enforcement, along with consistent and clear communication (particularly regarding balconies, patios and outdoor spaces) will encourage compliance.


[Contrast-enhanced spectral mammography. Experience in 465 examinations].
Contrast-enhanced spectral mammography or "contrast mammography" has a better cost effectiveness than breast magnetic resonance for confirmation of suspicious lesions detected on breast screening programs. To report the experience of a single center in Santiago. All patients referred for contrast mammography between July 2015 and October 2017 were studied. We recorded the patient risk factors for breast cancer. In 85 patients with suspicious lesions, biopsy results were available. We analyzed 465 contrast mammographies. The most common clinical indications were suspicion of cancer and previous inconclusive studies. Mass type lesions were detected in 33% of the studies. Non-mass-type lesions were observed in 10% of cases and findings compatible with papillomatosis in 2%. Fifty five percent of the studies had no visible lesions. In the 85 patients with a pathological study of the biopsy, the sensitivity of the contrast mammography was 100%, with a diagnostic accuracy of 85%, positive and negative predictive values of 82 and 100% respectively. Contrast mammography can be of great use for the assessment of patients with an altered conventional mammography, before indicating a magnetic resonance imaging or a percutaneous biopsy.


Response of human prostate cancer cells and tumors to combining PARP inhibition with ionizing radiation.
Radiation therapy remains a promising modality for curative treatment of localized prostate cancer, but dose-limiting toxicities significantly limit its effectiveness. Agents that enhance efficacy at lower radiation doses might have considerable value in increasing tumor control without compromising organ function. Here, we tested the hypothesis that the PARP inhibitor ABT-888 (veliparib) can enhance the response of prostate cancer cells and tumors to ionizing radiation (IR). Following exposure of DU-145 and PC-3 prostate cancer cell lines to the combination of 10 μmol/L ABT-888 and 6 Gy, we observed similar persistence between both cell lines of DNA damage foci and in vitro radiosensitization. We have previously observed that persistent DNA damage foci formed after ABT-888 plus IR efficiently promote accelerated cell senescence, but only PC-3 cells displayed the expected senescent response of G(2)-M arrest, induction of p21 and β-galactosidase expression, and accumulation as large flat cells. In turn, combining ABT-888 with 6 Gy resulted in delayed tumor regrowth compared with either agent alone only in PC-3 xenograft tumors, whereas DU-145 tumors continued to grow. By 7 days after treatment with ABT-888 plus IR, PC-3 tumors contained abundant senescent cells displaying persistent DNA damage foci, but no evidence of senescence was noted in the DU-145 tumors. That equivalent radiosensitization by ABT-888 plus IR in vitro failed to predict comparable results with tumors in vivo suggests that the efficacy of PARP inhibitors may partially depend on a competent senescence response to accumulated DNA damage.


A model of chemotherapy education for novice oncology nurses that supports a culture of safety.
Chemotherapy education at a mid-sized community hospital was redesigned to help novice oncology nurses improve patient safety and their own practice by implementing error prevention techniques during chemotherapy administration. Using a proactive approach with multidisciplinary participation and open communication, a systems analysis was conducted to identify potential chemotherapy errors. Then, chemotherapy processes were devised or strengthened to avoid errors. The project required a philosophical shift from error measurement to safety promotion.


Influence of brain histamine on growth hormone secretion induced by alpha-2-receptor activation.
The effects of histamine (HA) and of (R) alpha-methyl-histamine (alpha-MeHA; a drug that decreases histaminergic tone through activation of presynaptic H3 receptors) on growth hormone (GH) secretion induced by clonidine (CLO), an alpha 2-adrenergic agonist, were examined in rats. HA (0.1 mumol/rat intracerebroventricularly) and alpha-MeHA (10 mg/kg i.a.) were administered 5 min and 3 h, respectively, before CLO (0.25 mg/kg i.a.). Blood samples for hormone determination were drawn from freely moving rats at various times before and after drug treatment. To characterize the mechanisms underlying the inhibition of GH secretion by HA, the effect of HA on GH induced by CLO was examined in rats depleted of catecholamine by pretreatment (2 h before) with alpha-methyl-p-tyrosine (alpha-MpT, 200 mg/kg s.c.) and in rats depleted of somatostatin by pretreatment (4 h before) with cysteamine (300 mg/kg s.c.). HA completely suppressed the GH-releasing effect of CLO whereas alpha-MeHA significantly enhanced the GH response to CLO. Neither alpha-MpT nor cysteamine pretreatment modified the inhibitory activity of HA on GH secretion elicited by CLO. The present results indicate that the inhibitory activity of brain HA on GH release consequent to activation of central alpha 2-receptors does not involve a modulatory action on catecholaminergic or somatostatinergic neurons but probably other mechanisms like the modulation of neurons synthetizing GH-releasing hormone.


Witnessing cyberbullying and suicidal ideation among middle school students.
Suicide is a significant public health concern school-aged youth. Although a growing body of literature has documented the association between cyberbullying and suicidal ideation, and the mediating effect of internalizing symptoms, there is no research to date examining the impact of witnessing cyberbullying and suicidal ideation. To address this gap, we conducted a cross-sectional study with middle school students (N = 130). Students completed questionnaires assessing witnessing cyberbullying, witnessing school bullying, depression, anxiety, and suicidal ideation. We used structural equation modeling to test a mediational model in which we hypothesized internalizing symptoms would mediate the unique relationship between witnessing cyberbullying and suicidal ideation (i.e., controlling for witnessing school bullying). Results supported the mediational model, indicating the frequency of witnessing cyberbullying was positively related to internalizing symptoms, which in turn were related to higher levels of suicidal ideation. Findings suggest the importance of providing programs to support middle school students who witness cyberbullying to reduce the mental health risks (i.e., internalizing symptoms and suicidal ideation) associated with being a cyberbullying bystander.


Hysteroscopic Findings Related with the Assessment and Treatment of Uterine Florid Cystic Endosalpingiosis: A Case Report and Review of All the Published Cases.
Endosalpingiosis is a rare benign condition characterized by the presence of tubal epithelium outside the Fallopian tube. The clinical presentation of endosalpingiosis is nonspecific, and the diagnosis is typically incidental in women undergoing surgery for pelvic pain, infertility, urinary symptoms, or a pelvic mass. It can only be confirmed with histopathological examination. We report the first case of uterine florid cystic endosalpingiosis, with unusual hysteroscopic findings. We reviewed all the published cases of uterine florid cystic endosalpingiosis and their clinical presentation including hysteroscopic characteristics. It is a rare benign condition, with only 32 cases described in the literature. This is the first hysteroscopic description of this condition to be made. The patient first underwent a hysteroscopy and a leiomyoma resection when she was 51 years old. At 55, she went through another hysteroscopy, and a polypoid lesion was excised. A third hysteroscopy, one year later, revealed a new polypoid lesion in a similar location. After the initial incisions, this polypoid lesion disappeared. By decreasing the intrauterine pressure, it became visible again, corresponding histologically to an endometrial polyp with tubal metaplasia. At last, she underwent a laparoscopic hysterectomy with a final histopathological diagnosis of uterine florid cystic endosalpingiosis. Florid cystic endosalpingiosis is a rare condition that may be associated with several bizarre hysteroscopic findings.


Association Between 3 Doses of mRNA COVID-19 Vaccine and Symptomatic Infection Caused by the SARS-CoV-2 Omicron and Delta Variants.
Assessing COVID-19 vaccine performance against the rapidly spreading SARS-CoV-2 Omicron variant is critical to inform public health guidance. To estimate the association between receipt of 3 doses of Pfizer-BioNTech BNT162b2 or Moderna mRNA-1273 vaccine and symptomatic SARS-CoV-2 infection, stratified by variant (Omicron and Delta). A test-negative case-control analysis among adults 18 years or older with COVID-like illness tested December 10, 2021, through January 1, 2022, by a national pharmacy-based testing program (4666 COVID-19 testing sites across 49 US states). Three doses of mRNA COVID-19 vaccine (third dose ≥14 days before test and ≥6 months after second dose) vs unvaccinated and vs 2 doses 6 months or more before test (ie, eligible for a booster dose). Association between symptomatic SARS-CoV-2 infection (stratified by Omicron or Delta variants defined using S-gene target failure) and vaccination (3 doses vs unvaccinated and 3 doses vs 2 doses). Associations were measured with multivariable multinomial regression. Among cases, a secondary outcome was median cycle threshold values (inversely proportional to the amount of target nucleic acid present) for 3 viral genes, stratified by variant and vaccination status. Overall, 23 391 cases (13 098 Omicron; 10 293 Delta) and 46 764 controls were included (mean age, 40.3 [SD, 15.6] years; 42 050 [60.1%] women). Prior receipt of 3 mRNA vaccine doses was reported for 18.6% (n = 2441) of Omicron cases, 6.6% (n = 679) of Delta cases, and 39.7% (n = 18 587) of controls; prior receipt of 2 mRNA vaccine doses was reported for 55.3% (n = 7245), 44.4% (n = 4570), and 41.6% (n = 19 456), respectively; and being unvaccinated was reported for 26.0% (n = 3412), 49.0% (n = 5044), and 18.6% (n = 8721), respectively. The adjusted odds ratio for 3 doses vs unvaccinated was 0.33 (95% CI, 0.31-0.35) for Omicron and 0.065 (95% CI, 0.059-0.071) for Delta; for 3 vaccine doses vs 2 doses the adjusted odds ratio was 0.34 (95% CI, 0.32-0.36) for Omicron and 0.16 (95% CI, 0.14-0.17) for Delta. Median cycle threshold values were significantly higher in cases with 3 doses vs 2 doses for both Omicron and Delta (Omicron N gene: 19.35 vs 18.52; Omicron ORF1ab gene: 19.25 vs 18.40; Delta N gene: 19.07 vs 17.52; Delta ORF1ab gene: 18.70 vs 17.28; Delta S gene: 23.62 vs 20.24). Among individuals seeking testing for COVID-like illness in the US in December 2021, receipt of 3 doses of mRNA COVID-19 vaccine (compared with unvaccinated and with receipt of 2 doses) was less likely among cases with symptomatic SARS-CoV-2 infection compared with test-negative controls. These findings suggest that receipt of 3 doses of mRNA vaccine, relative to being unvaccinated and to receipt of 2 doses, was associated with protection against both the Omicron and Delta variants, although the higher odds ratios for Omicron suggest less protection for Omicron than for Delta.


Neoadjuvant chemotherapy of locally advanced breast cancer: predicting response with in vivo (1)H MR spectroscopy--a pilot study at 4 T.
To determine if changes in the concentration of choline-containing compounds (tCho) from before primary systemic therapy (PST) to within 24 hours after the first treatment enable prediction of clinical response in patients with locally advanced breast cancer. Sixteen women with biopsy-confirmed locally advanced breast cancer scheduled to undergo doxorubicin-based PST were recruited. Magnetic resonance (MR) imaging and spectroscopy were performed at 4 T prior to treatment, within 24 hours after the first dose, and after the fourth dose. Lesion size was assessed by using gadolinium-enhanced MR imaging. Lesion tCho concentration was quantified by using single-voxel hydrogen 1 MR spectroscopy. Statistical analysis was performed by using the Pearson correlation coefficient and the Wilcoxon rank sum test. Fourteen of 16 patients completed the protocol. In one patient, the level of tCho was not measurable because of unfavorable lesion morphology for MR spectroscopy voxel placement. Of the remaining 13 patients, four had inflammatory breast cancer, six had invasive ductal carcinoma, two had invasive lobular carcinoma, and one had mixed invasive ductal and lobular carcinoma. On the basis of the Response Evaluation Criteria in Solid Tumors, eight of 13 patients had an objective response and five had no response. The change in concentration of tCho from baseline to within 24 hours after the first dose of PST showed significant positive correlation with the change in lesion size (R = 0.79, P = .001). Change in tCho concentration within 24 hours after first dose was significantly different between patients with objective response and those with no response (P = .007). These results suggest that the change in tCho concentration between baseline and 24 hours after the first dose of PST can serve as an indicator for predicting clinical response to doxorubicin-based chemotherapy in locally advanced breast cancer.


First Record of Dothistroma pini on Pinus nigra in Switzerland.
The mitosporic ascomycetes Dothistroma septosporum s.s. (Dorog.) Morelet and D. pini Hulbary are closely related species (1) causing red band needle blight on Pinus spp. D. septosporum (teleomorph Mycosphaerella pini Rostr.) is considered as a cosmopolitan species, whereas D. pini (no teleomorph known) seems to have a more restricted distribution area. Detected in the United States on Pinus nigra for the first time, it was later found in Russia, Ukraine, Hungary, and France on different pine species (P. radiata, P. mugo, P. pallasiana) (3). In Switzerland, Dothistroma sp. (species not further determined) was recorded in 1989 for the first time and since then only damages on planted pines (mainly P. mugo and P. nigra) in urban areas were reported (R. Engesser, personal communication). In September 2012 and in April 2013, several planted mature trees and naturally regenerated young trees of P. nigra with Dothistroma needle blight were detected on a climatically mild forest site on limestone at the shore of Lake Walensee (47°07'48.0″ N, 9°13'54.4″ E, 420 m asl). In 2012, symptomatic needles were collected from the litter under one planted mature P. nigra tree and in 2013, symptomatic needles were collected from green twigs from a 2 m tall naturally regenerated P. nigra specimen. Conidiomata were frequently observed in the red bands but no conidia were detected. For fungal isolation, the surface of infected needles was shortly disinfected with 95% ethanol. The epi- and hypo-dermis covering the still closed conidiomata was removed and small tissue samples from the mesophyll (less than 0.5 mm length) were placed on malt extract agar (15 g/liter agar, 20 g/liter malt extract) amended by 50 mg/liter oxytetracycline. Conidia were observed after one year at 4°C in the resulting pure colonies (3 to 4 cm diameter on malt extract agar medium). The conidia formed by strain OH_120923_2_1_1 (KJ878557 = D. pini) were hyaline, smooth, thin-walled, 2- to 4-celled, and 31.6 (22 to 37) × 2.8 (2 to 3.5) μm. While conidial morphology of both Dothistroma species overlap, DNA was extracted and the internal transcribed spacer (ITS) region (primers ITS 1 and ITS 4) sequenced (KJ878557 to 81). From the 25 obtained ITS sequences, seven were identical with AY808275 (D. septosporum from P. radiata, South Africa, CMW 684), three were identical with AY808302 (D. pini from P. nigra, Michigan, CMW 10951), and 15 were identical with DQ926964 (D. pini from P. pallasiana, Ukraine, CMW 23767). The North American and Ukrainian D. pini sequences (AY808302 and DQ926964) showed only 1 bp difference. In addition, mating type genes were amplified using the method described by Groenewald et al. (2) for D. pini and scored using gel electrophoresis. Analyses showed that both D. pini ITS-sequence variants (e.g., KJ878557 and KJ878558) and both mating types were sometimes present in the same needle. In two cases, both mating types and ITS-sequence variants were also present within the same lesion. Interestingly, D. pini and D. septosporum were found on the same tree but not on the same needles. This is the first report of D. pini in Switzerland. Although symptoms of red band needle blight (species not determined) were repeatedly observed on this site during the last 20 years, the disease level always remained low and no tree mortality was noted. However, due to the presence of two ITS-sequence variants and both mating types, the incidence of D. pini in Switzerland deserves attention. References: (1) I. Barnes et al. Stud. Mycol. 50:551, 2004. (2) M. Groenewald et al. Phytopathology 97:825, 2007. (3) D. Piou and R. Ioos. Plant Dis. 98:841, 2014.


Clinical Utilization Pattern of Liquid Biopsies (LB) to Detect Actionable Driver Mutations, Guide Treatment Decisions and Monitor Disease Burden During Treatment of 33 Metastatic Colorectal Cancer (mCRC) Patients (pts) at a Fox Chase Cancer Center GI Oncology Subspecialty Clinic.
Background: Liquid biopsy (LB) captures dynamic genomic alterations (alts) across metastatic colorectal cancer (mCRC) therapy and may complement tissue biopsy (TB). We sought to describe the utility of LB and better understand mCRC biology during therapy. Methods: Thirty-three patients (pts) with mCRC underwent LB. We used permutation-based t-tests to assess associations between alts, and clinical variables and used Kendall's tau to measure correlations. Results: Of 33 pts, 15 were women; 22 had colon, and the rest rectal cancer. Pts received a median of two lines of therapy before LB. Nineteen pts had limited testing on TB (RAS/RAF/TP53/APC), 11 extended NGS, and 3 no TB. Maxpct and alts correlated with CEA (p < 0.001, respectively). In 3/5 pts with serial LB, CEA correlated with maxpct trend, and CT tumor burden. In 6 pts, mutant RAS was seen in LB and not TB; 5/6 had received anti-EGFR therapy prior to LB, suggesting RAS alts developed post-therapy. In two pts RAS-mutated by TB, no RAS alts were detected on LB; these pts had low disease burden on CT at time of LB that also did not reveal APC or TP53 alts. In six patients who were KRAS wt based on TB, post anti-EGFR LB revealed subclonal KRAS mutations, likely a treatment effect. The median number of alts was higher post anti-EGFR LB (n = 12) vs. anti-EGFR naïve LB (n = 22) (9.5 vs. 5.5, p = 0.059) but not statistically significant. More alts were also noted in post anti-EGFR therapy LB vs. KRAS wt anti-EGFR-naïve LB (n = 6) (9.5 vs. 5) among patients with KRAS wild-type tumors, although the difference was not significant (p = 0.182). Conclusions: LB across mCRC therapy detects driver mutations, monitors disease burden, and identifies sub-clonal alts that reflect drug resistance, tumor evolution, and heterogeneity. Interpretation of LB results is impacted by clinical context.


SARS-CoV-2 and SARS-CoV Spike-Mediated Cell-Cell Fusion Differ in Their Requirements for Receptor Expression and Proteolytic Activation.
Severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) infects cells through interaction of its spike protein (SARS2-S) with angiotensin-converting enzyme 2 (ACE2) and activation by proteases, in particular transmembrane protease serine 2 (TMPRSS2). Viruses can also spread through fusion of infected with uninfected cells. We compared the requirements of ACE2 expression, proteolytic activation, and sensitivity to inhibitors for SARS2-S-mediated and SARS-CoV-S (SARS1-S)-mediated cell-cell fusion. SARS2-S-driven fusion was moderately increased by TMPRSS2 and strongly by ACE2, while SARS1-S-driven fusion was strongly increased by TMPRSS2 and less so by ACE2 expression. In contrast to that of SARS1-S, SARS2-S-mediated cell-cell fusion was efficiently activated by batimastat-sensitive metalloproteases. Mutation of the S1/S2 proteolytic cleavage site reduced effector cell-target cell fusion when ACE2 or TMPRSS2 was limiting and rendered SARS2-S-driven cell-cell fusion more dependent on TMPRSS2. When both ACE2 and TMPRSS2 were abundant, initial target cell-effector cell fusion was unaltered compared to that of wild-type (wt) SARS2-S, but syncytia remained smaller. Mutation of the S2 cleavage (S2') site specifically abrogated activation by TMPRSS2 for both cell-cell fusion and SARS2-S-driven pseudoparticle entry but still allowed for activation by metalloproteases for cell-cell fusion and by cathepsins for particle entry. Finally, we found that the TMPRSS2 inhibitor bromhexine, unlike the inhibitor camostat, was unable to reduce TMPRSS2-activated cell-cell fusion by SARS1-S and SARS2-S. Paradoxically, bromhexine enhanced cell-cell fusion in the presence of TMPRSS2, while its metabolite ambroxol exhibited inhibitory activity under some conditions. On Calu-3 lung cells, ambroxol weakly inhibited SARS2-S-driven lentiviral pseudoparticle entry, and both substances exhibited a dose-dependent trend toward weak inhibition of authentic SARS-CoV-2.IMPORTANCE Cell-cell fusion allows viruses to infect neighboring cells without the need to produce free virus and contributes to tissue damage by creating virus-infected syncytia. Our results demonstrate that the S2' cleavage site is essential for activation by TMPRSS2 and unravel important differences between SARS-CoV and SARS-CoV-2, among those, greater dependence of SARS-CoV-2 on ACE2 expression and activation by metalloproteases for cell-cell fusion. Bromhexine, reportedly an inhibitor of TMPRSS2, is currently being tested in clinical trials against coronavirus disease 2019. Our results indicate that bromhexine enhances fusion under some conditions. We therefore caution against the use of bromhexine in high dosages until its effects on SARS-CoV-2 spike activation are better understood. The related compound ambroxol, which similarly to bromhexine is clinically used as an expectorant, did not exhibit activating effects on cell-cell fusion. Both compounds exhibited weak inhibitory activity against SARS-CoV-2 infection at high concentrations, which might be clinically attainable for ambroxol.


Pseudotetrahedral Zn(II)-(R or S)-dihalogen-salicylaldiminato complexes with Λ- or Δ-chirality induction at-metal.
Reactions of enantiopure (S or R)-N-1-(phenyl)ethyl-2,4-X1,X2-salicylaldimine (S-H or R-H; X1, X2 = dihalogen) with Zn(II)-nitrate give bis[(S or R)-N-1-(phenyl)ethyl-2,4-X1,X2-salicylaldiminato-κ2N,O]-zinc(II), (Δ-ZnS or Λ-ZnR) with Δ/Λ-chirality induction at-metal in the C2-symmetric molecules. EI-mass spectra show parent ion peaks. X-ray structures indicate that two phenolate-oxygen and two imine-nitrogen atoms from two molecules of the Schiff bases coordinate to the Zn(II) ion in a pseudotetrahedral geometry. Structural analyses give evidence that the S- or R-ligand chirality gives only one diastereomer Δ-ZnS or Λ-ZnR in an enantiopure crystal. Gas-phase optimized structures suggest that the Δ-ZnS or Λ-ZnR diastereomers are slightly more stable than Λ-ZnS or Δ-ZnR by 1-2 kcal mol-1. The intramolecular interactions were analyzed with the Independent Gradient Model (IGM) using the program Multiwfn on the optimized structures and also indicate the diastereomeric preference of Δ-ZnS1 over Λ-ZnS1 (or Λ-ZnR1 over Δ-ZnR1). Variable time and temperature 1H NMR spectra support the presence of only one diastereomer Λ-ZnR or Δ-ZnS in the bulk samples, backed by the simulated spectra which exhibit no diastereomerization in solution. In contrast, the reported Zn(II)-(R or S)-salicylaldiminato/naphthaldiminato complexes show a diastereomeric mixture of both Δ- and Λ-forms and a Δ ⇄ Λ equilibrium in solution. Electronic circular dichroism (ECD) spectra in solution display expected mirror-image relationships for the (S or R)-Schiff base ligands and the (S or R)-ligated complexes. Combined analyses of experimental and simulated ECD spectra further support the notion of diastereomeric excess of Δ-ZnS or Λ-ZnR in solution. The overall results thus suggest the preservation of chirality at-zinc induced by S- or R-ligands in a solid or solution. Supramolecular packing analyses explore different kinds of intermolecular interactions with the strongest one for X⋯O. Only the halogen atom in the para position is involved in these interactions with Br⋯O > Cl⋯O. Hirshfeld surface analyses also support these interactions between two molecules at a distance shorter than the sum of the vdW radii. Comparison of the experimental and simulated PXRD patterns from the single-crystal X-ray structures shows a good matching and confirms the phase purity of the bulk samples.


Smad3 potentiates transforming growth factor beta (TGFbeta )-induced apoptosis and expression of the BH3-only protein Bim in WEHI 231 B lymphocytes.
Transforming growth factor-beta (TGFbeta) is a potent growth inhibitor and inducer of apoptosis in B lymphocytes and is essential for immune regulation and maintenance of self-tolerance. Here we show that exogenous overexpression of Smad3 potentiates TGFbeta-induced apoptosis and expression of the pro-apoptotic protein Bim in WEHI 231 B lymphocytes. Overexpression of dominant-negative forms of Smad3 abrogate these TGFbeta-induced responses. We also demonstrate that TGFbeta induces Bim protein expression concomitant with its induction of apoptosis in the mouse progenitor B lymphocyte cell line, Ba/F3. Enhanced expression of Bim protein induced by TGFbeta is associated with an increased association of Bim with Bcl-2 and a concomitant loss of mitochondrial membrane potential. Furthermore, we find that the anti-apoptotic effect of the pro-survival cytokine CD40 results in the abrogation of TGFbeta-mediated Bim induction. Our data provide the first evidence of Bim expression levels that are increased by the addition of a pro-apoptotic cytokine, TGFbeta, and also suggest that the TGFbeta-specific transcription factor Smad3 plays a role in mediating Bim expression levels and apoptosis.


Lack of consensus on corneal abrasion management: results of a national survey.
Our objective was to determine the practice patterns of Canadian emergency physicians with respect to the management of traumatic corneal abrasions. After developing our instrument and pilot testing it on a sample of emergency residents, we randomly surveyed 470 members of the Canadian Association of Emergency Physicians, using a modified Dillman technique. We distributed a pre-notification letter, an 18-item survey, and appropriate follow-up surveys to non-responders. Those members with an email address (n = 400) received a Web-based survey, and those without (n = 70) received a survey by post. The survey focused on the indications and utilization of analgesics (oral and topical), cycloplegics, eye patches and topical antibiotics. Our response rate was 64% (301/470), and the median age of respondents was 38 years. Most (77.7%) were male, 71.8% were full-time emergency physicians, 76.5% were emergency medicine certified, and 64.4% practised in teaching hospitals. Pain management preferences (offered usually or always) included oral analgesics (82.1%), cycloplegics (65.1%) and topical non-steroidal anti-inflammatory drugs (NSAIDs) (52.8%). Only 21.6% of respondents performed patching, and most (71.2%) prescribed topical antibiotics, particularly for contact lens wearers and patients with ocular foreign bodies. Two-thirds of the respondents provided tetanus toxoid if a foreign body was present, and 46.2% did so even if a foreign body was not present. Most respondents (88.0%) routinely arranged follow-up. This national survey of emergency physicians demonstrates a lack of consensus on the management of traumatic corneal abrasions. Further study is indicated to determine the optimal treatment, particularly regarding the use of topical NSAIDs.


Temporally unstable recurrence of earthquakes due to breaks in fractal scaling.
Observed sequences of large earthquakes are not consistent in either recurrence time or energy release; long-term prediction has been impossible even in areas, such as Parkfield, with well-defined recurrence intervals. The seismic gap hypothesis, which predicts characteristic earthquakes in areas of the circum-Pacific belt that have not produced recent great earthquakes, has also failed to predict the observed clustering of high-energy events. Models in which fractal scaling is broken at high magnitude predict that characteristic events and recurrence behavior will be unstable in time. The central predictions of these models are supported by recent observations at Landers and Big Bear in California.


Allosteric regulation of epidermal growth factor (EGF) receptor ligand binding by tyrosine kinase inhibitors.
The epidermal growth factor (EGF) receptor is a classical receptor tyrosine kinase with an extracellular ligand-binding domain and an intracellular kinase domain. Mutations in the EGF receptor have been shown to drive uncontrolled cell growth and are associated with a number of different tumors. Two different types of ATP-competitive EGF receptor tyrosine kinase inhibitors have been identified that bind to either the active (type I) or inactive (type II) conformation of the kinase domain. Despite the fact that both types of inhibitors block tyrosine kinase activity, they exhibit differential efficacies in different tumor types. Here, we show that in addition to inhibiting kinase activity, these inhibitors allosterically modulate ligand binding. Our data suggest that the conformations of the EGF receptor extracellular domain and intracellular kinase domain are coupled and that these conformations exist in equilibrium. Allosteric regulators, such as the small-molecule tyrosine kinase inhibitors, as well as mutations in the EGF receptor itself, shift the conformational equilibrium among the active and inactive species, leading to changes in EGF receptor-binding affinity. Our studies also reveal unexpected positive cooperativity between EGF receptor subunits in dimers formed in the presence of type II inhibitors. These findings indicate that there is strong functional coupling between the intracellular and extracellular domains of this receptor. Such coupling may impact the therapeutic synergy between small-molecule tyrosine kinase inhibitors and monoclonal antibodies in vivo.


COL11A1-Driven Epithelial-Mesenchymal Transition and Stemness of Pancreatic Cancer Cells Induce Cell Migration and Invasion by Modulating the AKT/GSK-3β/Snail Pathway.
Collagen type XI α1 (COL11A1) is associated with tumorigenesis and development in many human malignancies. Previous reports indicate that COL11A1 may be a significant diagnostic marker for pancreatic ductal adenocarcinoma (PDAC); however, its biological role in PDAC progression remains unclear. In this study, we investigated the influence of COL11A1 on the invasion and migration abilities of pancreatic cancer cells and explored its potential molecular mechanisms. Cell migration and invasion were assessed using Transwell assays in pancreatic cancer cells transfected with siCOL11A1 and pCNV3-COL11A1 plasmids. The protein and mRNA expression levels of N-cadherin, E-cadherin, Vimentin, cluster of differentiation (CD)-24, CD44, serine-threonine kinase (AKT), glycogen synthase kinase (GSK)-3β, phospho (p)-AKTSer473, p-GSK-3βSer9, and Snail were analyzed using Western blotting and real-time polymerase chain reaction (PCR). The effect of COL11A1 on cell stemness was tested using flow cytometry and clone formation assays. These results demonstrated that COL11A1 significantly promoted the invasion and migration abilities of PDAC cells. Furthermore, COL11A1 facilitated the occurrence of epithelial-mesenchymal transition (EMT) and cell stemness by upregulating the expression levels of p-AKTSer473, p-GSK-3βSer9, and Snail. This study suggests that the activation of the AKT/GSK-3β/Snail signaling pathway induced by COL11A1 plays a major role in the progression of PDAC. Therefore, COL11A1 could serve as a potential target for PDAC treatment.


Aberrant functional connectivity between reward and inhibitory control networks in pre-adolescent binge eating disorder.
Behavioral features of binge eating disorder (BED) suggest abnormalities in reward and inhibitory control. Studies of adult populations suggest functional abnormalities in reward and inhibitory control networks. Despite behavioral markers often developing in children, the neurobiology of pediatric BED remains unstudied. 58 pre-adolescent children (aged 9-10-years) with BED (mBMI = 25.05; s.d. = 5.40) and 66 age, BMI and developmentally matched control children (mBMI = 25.78; s.d. = 0.33) were extracted from the 3.0 baseline (Year 0) release of the Adolescent Brain Cognitive Development (ABCD) Study. We investigated group differences in resting-state functional MRI functional connectivity (FC) within and between reward and inhibitory control networks. A seed-based approach was employed to assess nodes in the reward [orbitofrontal cortex (OFC), nucleus accumbens, amygdala] and inhibitory control [dorsolateral prefrontal cortex, anterior cingulate cortex (ACC)] networks via hypothesis-driven seed-to-seed analyses, and secondary seed-to-voxel analyses. Findings revealed reduced FC between the dlPFC and amygdala, and between the ACC and OFC in pre-adolescent children with BED, relative to controls. These findings indicating aberrant connectivity between nodes of inhibitory control and reward networks were corroborated by the whole-brain FC analyses. Early-onset BED may be characterized by diffuse abnormalities in the functional synergy between reward and cognitive control networks, without perturbations within reward and inhibitory control networks, respectively. The decreased capacity to regulate a reward-driven pursuit of hedonic foods, which is characteristic of BED, may in part, rest on this dysconnectivity between reward and inhibitory control networks.


Heterodimer Binding Scaffolds Recognition via the Analysis of Kinetically Hot Residues.
Physical interactions between proteins are often difficult to decipher. The aim of this paper is to present an algorithm that is designed to recognize binding patches and supporting structural scaffolds of interacting heterodimer proteins using the Gaussian Network Model (GNM). The recognition is based on the (self) adjustable identification of kinetically hot residues and their connection to possible binding scaffolds. The kinetically hot residues are residues with the lowest entropy, i.e., the highest contribution to the weighted sum of the fastest modes per chain extracted via GNM. The algorithm adjusts the number of fast modes in the GNM's weighted sum calculation using the ratio of predicted and expected numbers of target residues (contact and the neighboring first-layer residues). This approach produces very good results when applied to dimers with high protein sequence length ratios. The protocol's ability to recognize near native decoys was compared to the ability of the residue-level statistical potential of Lu and Skolnick using the Sternberg and Vakser decoy dimers sets. The statistical potential produced better overall results, but in a number of cases its predicting ability was comparable, or even inferior, to the prediction ability of the adjustable GNM approach. The results presented in this paper suggest that in heterodimers at least one protein has interacting scaffold determined by the immovable, kinetically hot residues. In many cases, interacting proteins (especially if being of noticeably different sizes) either behave as a rigid lock and key or, presumably, exhibit the opposite dynamic behavior. While the binding surface of one protein is rigid and stable, its partner's interacting scaffold is more flexible and adaptable.


Healthcare worker experiences with Option B+ for prevention of mother-to-child HIV transmission in eSwatini: findings from a two-year follow-up study.
Prevention of mother-to-child transmission (PMTCT) across sub-Saharan Africa has rapidly shifted towards Option B+, an approach in which all HIV+ pregnant and breastfeeding women initiate lifelong antiretroviral therapy (ART) independent of CD4+ count. Healthcare workers (HCW) are critical to the success of Option B+, yet little is known regarding HCW acceptability of Option B+, particularly over time. Ten health facilities in the Manzini and Lubombo regions of eSwatini transitioned from Option A to Option B+ between 2013 and 2014 as part of the Safe Generations study examining PMTCT retention. Fifty HCWs (5 per facility) completed questionnaires assessing feasibility and acceptability: (1) prior to transitioning to Option B+, (2) two months post transition, and (3) approximately 2 years post Option B+ transition. This analysis describes HCW perceptions and experiences two years after transitioning to Option B+. Two years after transition, 80% of HCWs surveyed reported that Option B+ was easy for HCWs, noting that it was particularly easy to explain and coordinate. Immediate ART initiation also reduced delays by eliminating need for laboratory tests prior to ART initiation. Additionally, HCWs reported ease of patient follow-up (58%), documentation (56%), and counseling (58%) under Option B+. Findings also indicate that a majority of HCWs reported that their workloads increased under Option B+. Sixty-eight percent of HCWs at two years post-transition reported more work under Option B+, specifically noting increased involvement in adherence counseling, prescribing/monitoring medications, and appointment scheduling/tracking. Some HCWs attributed their higher workloads to increased client loads, now that all HIV-positive women were initiated on ART. New barriers to patient uptake, and issues related to retention, adherence, and follow-up were also noted as challenges face by HCW when implementing Option B+. Overall, HCWs found Option B+ to be acceptable and feasible while providing critical insights into the practical issues of universal ART. Further strengthening of the healthcare system may be necessary to alleviate worker burden and to ensure effective monitoring of client retention and adherence. HCW perceptions and experiences with Option B+ should be considered more broadly as countries implement Option B+ and consider universal treatment for all HIV+ individuals. http://clinicaltrials.gov NCT01891799 , registered on July 3, 2013.


A multi-disciplinary approach to the diagnosis and management of allergic diseases: An EAACI Task Force.
Guidelines for management of patients with allergic conditions are available, but the added value of nurses, allied healthcare professionals (AHPs), and general practitioners (GPs), in the management of allergic disease, has not been fully clarified. The European Academy of Allergy and Clinical Immunology (EAACI) appointed a task force to explore this issue. To investigate the added value of nurses, AHPs, and GPs in management of allergic diseases, in an integrated model of care. A search was made of peer-reviewed literature published between 2010 and December 2020 (Cochrane Library, PubMed, and CINAHL) on the involvement of the various specific healthcare providers (HCPs) in the management of allergic diseases. Facilitative models of care for patients with allergies can be achieved if HCP collaborates in the diagnosis and management. Working in multi-disciplinary teams (MDT) can increase patients' understanding of the disease, adherence to treatment, self-care capabilities, and ultimately improve quality of life. The MDT competencies and procedures can be improved and enhanced in a climate of mutual respect and shared values, and with inclusion of patients in the planning of care. Patient-centered communication among HCPs and emphasis on the added value of each profession can create an effective integrated model of care for patients with allergic diseases. Nurses, AHPs, and GPs, both individually and in collaboration, can contribute to the improvement of the management of patients with allergic disease. The interaction between the HCPs and the patients themselves can ensure maximum support for people with allergies.


Regulation of myogenesis by fibroblast growth factors requires beta-gamma subunits of pertussis toxin-sensitive G proteins.
Terminal differentiation of skeletal muscle cells in culture is inhibited by a number of different growth factors whose subsequent intracellular signaling events are poorly understood. In this study, we have investigated the role of heterotrimeric G proteins in mediating fibroblast growth factor (FGF)-dependent signals that regulate myogenic differentiation. Pertussis toxin, which ADP-ribosylates and inactivates susceptible G proteins, promotes terminal differentiation in the presence of FGF-2, suggesting that Galpha or Gbeta gamma subunits or both are involved in transducing the FGF-dependent signal(s) that inhibits myogenesis. We found that Gbetagamma subunits are likely to be involved since the expression of the C terminus of beta-adrenergic receptor kinase 1, a Gbetagamma subunit-sequestering agent, promotes differentiation in the presence of FGF-2, and expression of the free Gbeta gamma dimer can replace FGF-2, rescuing cells from pertussis toxin-induced differentiation. Addition of pertussis toxin also blocked FGF-2-mediated activation of mitogen-activated protein kinases (MAPKs). Ectopic expression of dominant active mutants in the Ras/MAPK pathway rescued cells from pertussis toxin-induced terminal differentiation, suggesting that the Gbeta gamma subunits act upstream of the Ras/MAPK pathway. It is unlikely that the pertussis toxin-sensitive pathway is activated by other, as yet unidentified FGF receptors since PDGF (platelet-derived growth factor)-stimulated MM14 cells expressing a chimeric receptor containing the FGF receptor-1 intracellular domain and the PDGF receptor extracellular domain were sensitive to pertussis toxin. Our data suggest that FGF-mediated signals involved in repression of myogenic differentiation are transduced by a pertussis toxin-sensitive G-protein-coupled mechanism. This signaling pathway requires the action of Gbeta gamma subunits and activation of MAPKs to repress skeletal muscle differentiation.