Infection of Dendritic Cells With Mycobacterium avium subspecies hominissuis Exhibits a Functionally Tolerogenic Phenotype in Response to Toll-Like Receptor Agonists via IL-10/Cox2/PGE2/EP2 Axis.
Mycobacterium avium subspecies hominissuis (MAH) is the most common agent causing nontuberculous mycobacterial disease in humans. It mainly causes chronic and slowly progressive pulmonary disease (PD), which requires a long-term treatment and allows opportunistic co-infection by common pulmonary pathogens such as Pseudomonas aeruginosa, Staphylococcus aureus, and Aspergillus spp., thereby resulting in alteration of host immune response. In the present study, we investigated the phenotypical and functional alterations of dendritic cells (DCs), a bridge antigen-presenting cell between innate and adaptive immunity, following MAH infection in response to various toll-like receptor (TLR) agonists mimicking co-infection conditions, along with subsequent T cell response. Interestingly, MAH-infected DCs produced interleukin (IL)-10 significantly and decreased the level of IL-12p70 in response to Poly I:C and LPS, although not so in response to Pam3CSK4, imiquimod, or CpG oligodeoxynucleotide, thereby indicating that the TLR3 and TLR4 agonists functionally altered MAH-infected DCs toward a tolerogenic phenotype. Moreover, IL-10-producing tolerogenic DCs were remarkably induced by MAH and P. aeruginosa co-infection. To precisely elucidate how these TLR agonists induce tolerogenic DCs upon MAH infection, we sought to clarify the major mechanisms involved, using LPS, which caused the greatest increase in IL-10 production by the TLR agonists. Increased IL-10 stimulated the creation of tolerogenic DCs by significantly reducing MHC class II expression and MHC class II-antigen presentation, eventually inhibiting CD4+ T cell proliferation, along with decreased IFN-γ and IL-2. The tolerogenic phenotypes of MAH/LPS-treated DCs were restored by anti-IL-10 neutralization, validating the induction of tolerogenicity by IL-10. Interestingly, IL-10-producing-tolerogenic DCs were observed after infection with live MAH, rather than with inactivated or dead MAH. In addition, TLR2-/- and TLR4-/- DCs confirmed the association of IL-10 production with TLR2 and TLR4 signaling; IL-10 production synergistically increased when both TLR4 and TLR2 were involved. Expression of Cox2 and PGE2 increased along with IL-10 while that of IL-10 was inhibited by their selective inhibitors celecoxib and anti-EP2 antibody, respectively. Thus, the tolerogenic phenotypes of MAH/LPS-treated DCs were proven to be induced by Cox-2/PGE2-dependent EP2 signaling as the main mechanism. These findings may provide important clues that the tolerogenic cascade in MAH-infected DCs induced by TLR 4 signaling can alter host immune response.


'A world of competing sorrows': A mixed methods analysis of media reports of children with cancer abandoning conventional treatment.
We aimed to provide health practitioners greater insight into the public perception of traditional and complementary medicine (T&CM) use. Our objectives were to identify news media reports of children abandoning conventional treatment for traditional and complementary medicine, analyze the thematic content of these news articles and estimate the tonality portrayed. LexisNexis and Factiva were searched for terms related to cancer, children and T&CM. Inclusion criteria were children less than 18 years, in curative phase of treatment who attempted to abandon conventional therapy for any traditional and complementary medicine use. A secondary search was performed in LexisNexis, Factiva and Google News Archive with the names of children in identified cases. Qualitative analysis of news media reports was completed using a grounded theory approach. Quantitative analysis of article sentiment was performed using a linear support vector machine. Seventeen cases occurring between 2002 and 2016 were included. Five main themes were identified: treatment as torture, power imbalances, rights of parents, evidence versus beliefs and the rights of Indigenous Peoples. Sentiment analysis revealed an overall negative tone, as demonstrated by 73% of the articles. A better understanding of factors that lead to abandonment of conventional therapy for traditional and complementary medicine as portrayed in the news media may help healthcare providers prevent the occurrence of these cases.


Dual orexin and MCH neuron-ablated mice display severe sleep attacks and cataplexy.
Orexin/hypocretin-producing and melanin-concentrating hormone-producing (MCH) neurons are co-extensive in the hypothalamus and project throughout the brain to regulate sleep/wakefulness. Ablation of orexin neurons decreases wakefulness and results in a narcolepsy-like phenotype, whereas ablation of MCH neurons increases wakefulness. Since it is unclear how orexin and MCH neurons interact to regulate sleep/wakefulness, we generated transgenic mice in which both orexin and MCH neurons could be ablated. Double-ablated mice exhibited increased wakefulness and decreased both rapid eye movement (REM) and non-REM (NREM) sleep. Double-ablated mice showed severe cataplexy compared with orexin neuron-ablated mice, suggesting that MCH neurons normally suppress cataplexy. Double-ablated mice also showed frequent sleep attacks with elevated spectral power in the delta and theta range, a unique state that we call 'delta-theta sleep'. Together, these results indicate a functional interaction between orexin and MCH neurons in vivo that suggests the synergistic involvement of these neuronal populations in the sleep/wakefulness cycle.


Allosteric inhibition of the epithelial Na+ channel through peptide binding at peripheral finger and thumb domains.
The epithelial Na(+) channel (ENaC) mediates the rate-limiting step in transepithelial Na(+) transport in the distal segments of the nephron and in the lung. ENaC subunits are cleaved by proteases, resulting in channel activation due to the release of inhibitory tracts. Peptides derived from these tracts inhibit channel activity. The mechanism by which these intrinsic inhibitory tracts reduce channel activity is unknown, as are the sites where these tracts interact with other residues within the channel. We performed site-directed mutagenesis in large portions of the predicted periphery of the extracellular region of the α subunit and measured the effect of mutations on an 8-residue inhibitory tract-derived peptide. Our data show that the inhibitory peptide likely binds to specific residues within the finger and thumb domains of ENaC. Pairwise interactions between the peptide and the channel were identified by double mutant cycle experiments. Our data suggest that the inhibitory peptide has a specific peptide orientation within its binding site. Extended to the intrinsic inhibitory tract, our data suggest that proteases activate ENaC by removing residues that bind at the finger-thumb domain interface.


Does Intrawound Vancomycin Application During Spine Surgery Create Vancomycin-Resistant Organism?
Surgical site infection (SSI) following spine surgery is a morbid and expensive complication. The use of intrawound vancomycin is emerging as a solution to reduce SSI. The development of vancomycin-resistant pathogens is an understandable concern. To determine the occurrence of vancomycin-resistant SSI in patients with and without use of intrawound vancomycin. Patients undergoing elective spine surgery were dichotomized based on whether intrawound vancomycin was applied. Outcome was occurrence of SSI requiring return to the operating room within postoperative 90 days. The intrawound culture and vancomycin minimal inhibitory concentrations (MIC) were reviewed. Analyses were conducted to compare the pathogen profile and MIC for vancomycin in patients who received vancomycin and those who did not. Of the total 2802 patients, 43% (n = 1215) had intrawound vancomycin application during the index surgery. The use of vancomycin was associated with significantly lower deep SSI rates (1.6% [n = 20] vs 2.5% [n = 40], P = .02). The occurrence of Staphylococcus aureus SSI was significantly lower in the patients who had application of intrawound vancomycin (32% vs 65%, P = .003). None of the patients who had application of intrawound vancomycin powder, and subsequently developed an S aureus SSI, demonstrated pathogens with resistance to vancomycin. All patients had MIC < 2 μg/mL, the vancomycin susceptibility threshold. The occurrence of gram-negative SSI (28% vs 7%) and culture negative fluid collection (16% vs 5%) was higher in the vancomycin cohort. The use of intrawound vancomycin during the index spine surgery was protective against SSI following spine surgery. The application of intrawound vancomycin during index surgery does not appear to create vancomycin-resistant organisms in the event of an SSI.


Experience with NONMEM: analysis of routine phenytoin clinical pharmacokinetic data.
NONMEM, a program package the produces the extended least squares estimates of population parameters for a nonlinear mixed-effect model, has been applied to two data sets from patients routinely receiving phenytoin. A general model for the data is proposed. The models used in previous, standard-method analyses of each data set are compared to the general model using NONMEM. The comparison involves two questions: The first asks whether the parameters estimated previously agree with NONMEM estimates when the original model is used. We find that for fixed-effect parameters they generally do, while for interindividual random-effect parameters the previous methods' estimates appear upward biased relative to NONMEM. Second, the original model per se is compared to the general model by comparing the best fit to each. The general model is clearly superior. NONMEM's ability to distinguish among models, and to precisely estimate their parameters from sparse individual data, is illustrated and verified.


Strengthening the feedback culture in a postgraduate residency program.
Feedback is defined as specific information presented to a learner that facilitates professional development through the process of reflection. Timely provision of constructive feedback to learner is important in optimizing the learning curve. The aim of the current study was to see the effectiveness of various interventions on feedback practices of faculty members. This is a quasi-experimental study (pre- and postdesign). It was conducted from November 2009 to March 2011 at The Aga Khan University, Pakistan. Faculty development workshops, allotment of specified feedback time, and restructuring of residency feedback forms were done as interventions. Data collection was done pre- and postintervention. Resident's and faculty satisfaction regarding the feedback process were evaluated using a prepiloted questionnaire. Paired t-test was applied to assess the effect of interventions on faculty and resident's satisfaction. The mean satisfaction scores of residents were significantly improved (P < 0.05). Pre- and postintervention faculty satisfaction score also demonstrated significant difference in overall satisfaction level, from 47.88 ± 13.92 to 63.40 ± 8.72 (P < 0.05). This study showed improved faculty engagement and satisfaction for the provision of feedback to the trainee resident. Strengthening this, culture requires continuous reinforcement, individualized feedback to the faculty members regarding their feedback practices, and continuing faculty development initiatives.


Cholera in Bahrain: epidemiological characteristics of an outbreak.
In the period 10 August 1978-23 January 1979, 913 culture-confirmed cases of cholera caused by Vibrio cholerae, biotype El Tor, serotype Ogawa, occurred in Bahrain. After discovery of the initial cases, others occurred sporadically, and the incidence reached a peak of 25-35 cases per day during the seventh week of the outbreak (16-22 September). The overall attack rate (27 per 10 000) was low and the outbreak subsided without mass immunization campaigns or rigorous border control of persons and imports. Investigation of 746 culture-confirmed cases that occurred in the period 10 August-13 October 1978, showed that cases occurred throughout most areas of the country and mainly affected infants, young children, and adult working-age males. Symptoms were very mild; fewer than 20% of patients required specific rehydration therapy. The highest attack rate (84 per 10 000) occurred in infants less than 1 year of age. No common vehicle or mode of transmission was identified. A matched-pair study of 35 cases and controls showed that adult cases were more likely than controls to have consumed food or beverage outside of the home before becoming ill. V. cholerae was isolated from stored drinking water in the houses of 8 cases but not from numerous samples of food and tap-water. It was presumed that cholera transmission occurred through a complex interaction of mild and asymptomatically infected persons with food, water, and the environment.


Levels of Apolipoprotein A1, B100 and Lipoprotein (a) in Controlled and Uncontrolled Diabetic Patients and in Non-Diabetic Healthy People.
Diabetes Mellitus (DM) is always a multifactorial metabolic disorder having a wide range of abnormalities in carbohydrate, lipid and protein metabolism. Dyslipidemia is a natural process of DM causing abnormal variations of different lipoproteins and it is one of the significant risk factors for Cardiovascular Disorder (CVD). There is a need to closely evaluate newer approaches in case of DM because even if dyslipidemia is treated, there is always a risk of CVDs in DM patients because of the hyperglycemia itself. So, lipid abnormalities should be assessed aggressively and treated as part of diabetes care. Apolipoprotein B100 (Apo B100), Apolipoprotein A1 (Apo A1) and Lipoprotein (a) {Lp(a)} are newer markers which are always welcome and necessary as many of the reported cases with normal conventional lipid profile have developed cardiac events. Study the correlation between glycemic control and the levels of Apo A1, Apo B100 and Lp(a). Total 56 patients of (DM) diagnosed on the basis of American Diabetic Association guidelines were recruited, out of which 28 were identified as uncontrolled-diabetic patients and remaining 28 as controlled-diabetics on the basis of Glycosylated HbA1c (HbA1c). The control group consisted of normal healthy 28 individuals. Apo B100, Apo A1 and Lp(a) along with traditional lipid profile, Fasting Blood Sugar (FBS) and HbA1c were estimated in all the subjects. Apo B100/Apo A1 ratio and Lp(a) levels showed highly significant difference (p-value <0.001) between uncontrolled diabetics, controlled diabetics and healthy Controls. Apo B100/Apo A1 ratio and Lp(a) showed significant positive correlations with HbA1c (r= 0.494, p <0.0001) and with each other. Apo B100/Apo A1 ratio and Lp(a) show a highly significant positive relationship with glucose tolerance of the patients as reflected in the HbA1c values. If proper glycemic control is maintained, the levels of Apo B100/Apo A1 ratio and Lp(a) can be controlled as reflected by the lower levels of these parameters observed in controlled diabetics in comparison to uncontrolled diabetics.


Origins of netropsin binding affinity and specificity: correlations of thermodynamic and structural data.
We report complete thermodynamic profiles for netropsin binding to an oligomeric and to several polymeric DNA host duplexes. These data allow us to reach the following conclusions: netropsin binding by deep penetration into the minor groove is overwhelmingly enthalpy driven and exhibits a very high binding affinity (K approximately 10(9) at 25 degrees C); deep penetration into the minor groove is required to form those drug-DNA interactions responsible for the enthalpy-driven high binding affinity of netropsin; I-C base pairs form binding sites for netropsin that thermodynamically are equivalent to those formed by A-T base pairs; the positive binding entropies reflect entropic contributions from molecular events other than just water spine disruption; the thermodynamic binding data primarily reflect local netropsin-DNA interactions rather than long-range binding-induced conformational changes at regions distant from the binding site; the enhanced binding affinity associated with deep penetration of netropsin into the minor groove does not result from more favorable electrostatic interactions; the binding of netropsin to the central AATT core of the decamer duplex [d(GCGAATTCGC)]2 is thermodynamically modeled best by netropsin binding to the poly[d(AT)].poly[d(AT)] duplex rather than the poly(dA).poly(dT) duplex. We propose correlations between our thermodynamic data and specific molecular interactions defined by NMR and x-ray structural studies on similar and identical drug-DNA complexes.


Intestinal full-thickness needle-core biopsy via laparotomy is safe, rapid, and effective and less invasive than standard incisional biopsy in dogs and cats.
To describe the intestinal full-thickness needle-core biopsy technique via abdominal laparotomy outcomes and compare the histopathological and immunohistochemical diagnosis with standard incisional intestinal biopsy technique in dogs and cats. 3 dogs and 17 cats. Client-owned dogs and cats were prospectively enrolled if intestinal full-thickness biopsies were indicated for the diagnosis of diffuse chronic intestinal diseases following ultrasonography. The study period extended from June 2021 to December 2022. All animals underwent intestinal biopsies with both techniques (needle-core biopsy and standard incisional biopsy) via abdominal laparotomy. Data collected included clinical signs, biopsy collection times, complications, and histopathologic and immunohistochemical findings. A minimum follow-up of 14 days was required. The main clinical sign at presentation was chronic vomiting (65%). Mean needle-core biopsy collection time (262 seconds) was significantly shorter than standard incisional biopsy collection time (599 seconds; P < .000001). The incidence of minor complications was 10% (inflammation of the skin surgical site secondary to licking). One catastrophic complication occurred on a standard incisional biopsy site in 1 cat in a context of bile peritonitis (5% of all cases). There were no complications associated with the needle-core biopsy. All but 1 cat were discharged, with a median of 2 days (range, 1 to 4 days) after surgery. The diagnoses resulting from both techniques were 100% concordant for the distinction between inflammatory bowel disease and intestinal lymphoma via histopathology and immunochemistry. Needle-core biopsy is safe, rapid, and effective and is less invasive than standard incisional biopsy.


[Therapy of idiopathic sterility].
During the period 1996-2000, we investigated 900 couples with infertility. In 71 (7.9%) couples no cause of sterility was revealed. These patients were divided according to age, parity and infertility duration. We established the cumulative conception rate and cycle fecundity after three months without therapy, after three cycles of intrauterine insemination and after three cycles of in vitro fertilization (eventually intracytoplasmic sperm injection). Among patients with less than three years of infertility, cumulative conception rate was 34.8% after three months without therapy, 27.3% after three cycles of intrauterine insemination and 66.7% after three cycles of in vitro fertilization. In patients with infertility longer than three years, cumulative conception rate without therapy was 12.5%, with intrauterine insemination 16.1% and with in vitro fertilization 40.5%. Differences regarding the age of patients were significant. Based on these findings, we proposed an optimal therapy regimen: three months therapy delay is desirable in patients under 30 years of age with infertility duration under three years. If there is no pregnancy in that period, intrauterine insemination is performed in three to four cycles. In case of intrauterine insemination failure in these women, in vitro fertilization and embryo transfer should be applied. In the group of women over 30 years of age, with infertility longer than three years, the possibility for occurrence of pregnancy with intrauterine insemination is slightly increased. If more than 4 follicles develop during preparations for intrauterine insemination, it is advisable to change the course to in vitro fertilization. In women older than 35, with primary or secondary infertility, in vitro fertilization should be performed from the very beginning.


Parkinson's disease: implications for nutritional care.
Parkinson's Disease (PD) is a chronic, progressive, neurodegenerative disease. People with PD are particularly susceptible to weight loss and malnutrition. Involuntary movements associated with PD result in increased energy expenditure, while both disease symptoms and medication side-effects can limit food intake. In addition, patients with the disease may choose to follow unconventional nutritional therapies that exacerbate malnutrition. Dietitians play a key role in helping patients with PD to optimize their nutritional status and manage various nutrition-related symptoms and medication side-effects. To assume this role, dietitians need to have current knowledge about PD and its nutritional consequences, as well as strategies for managing a variety of nutrition-related symptoms.


S32212, a novel serotonin type 2C receptor inverse agonist/α2-adrenoceptor antagonist and potential antidepressant: I. A mechanistic characterization.
Although most antidepressants suppress serotonin (5-HT) and/or noradrenaline reuptake, blockade of 5-HT(2C) receptors and α(2)-adrenoceptors likewise enhances monoaminergic transmission. These sites are targeted by the urea derivative N- [4-methoxy-3-(4-methylpiperazin-1-yl)phenyl]-1,2-dihydro-3-H-benzo[e]indole-3-carboxamide (S32212). S32212 was devoid of affinity for monoamine reuptake sites, yet displayed pronounced affinity (pK(i), 8.2) for constitutively active human 5-HT(2CINI) (h5-HT(2CINI)) receptors, behaving as an inverse agonist in reducing basal Gα(q) activation, [(3)H]inositol-phosphate production, and the spontaneous association of h5-HT(2CINI)-Renilla luciferase receptors with β-arrestin2-yellow fluorescent protein. Furthermore, upon 18-h pretreatment, S32212 enhanced the plasma membrane expression of h5-HT(2CINI) receptors as visualized by confocal microscopy and quantified by enzyme-linked immunosorbent assay. Its actions were prevented by the neutral antagonist 6-chloro-5-methyl-N-[6-(2-methylpyridin-3-yloxy)pyridin-3-yl]indoline-1-carboxamide (SB242,084), which also impeded the induction by long-term exposure to S32212 of otherwise absent Ca(2+) mobilization in mouse cortical neurones. In vivo, S32212 blunted the inhibitory influence of the 5-HT(2C) agonist 2-(3-chlorobenzyloxy)-6-(1-piperazinyl)pyrazine (CP809,101) on ventrotegmental dopaminergic neurones. S32212 also blocked 5-HT-induced Gα(q) and phospholipase C activation at the h5-HT(2A) and, less potently, h5-HT(2B) receptors and suppressed the discriminative stimulus properties of the 5-HT(2A) agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane in rats. S32212 manifested marked affinity for human α(2A)- (pK(i) 7.2), α(2B)- (pK(i) 8.2), and α(2C)- (pK(i) 7.4) adrenoceptors, at which it abolished noradrenaline-induced recruitment of Gα(i3), Gα(o), adenylyl cyclase, and extracellular-regulated kinase1/2. Moreover, S32212 dose-dependently abolished the discriminative stimulus effects of the α(2)-adrenoceptor agonist (S)-spiro[(1-oxa-2-amino-3-azacyclopent-2-ene)-4,2'-(1',2',3',4'-tetrahydronaphthalene)] (S18616). Finally, S32212 displayed negligible affinity for α(1A)-adrenoceptors, histamine H(1) receptors, and muscarinic M(1) receptors. In conclusion, S32212 behaves as an inverse agonist at h5-HT(2C) receptors and as an antagonist at human α(2)-adrenoceptors (and h5-HT(2A) receptors). Its promising profile in preclinical models potentially relevant to the treatment of depression is described in J Pharmacol Exp Ther 340:765-780, 2012.


Does the prognostic value of dobutamine stress echocardiography differ among different age groups?
Age is associated with reduced exercise capacity and greater prevalence of coronary artery disease. Whether the prognostic information obtained from dobutamine stress echocardiography (DSE), a stress test commonly used for patients unable to perform an exercise test, provides differential information based on age is not well known. We studied 6,655 consecutive patients referred for DSE. Patients were divided into 3 age groups: (1) <60 years (n = 1,389), (2) 60 to 74 years (n = 2,978), and (3) ≥75 years (n = 2,288). Mean follow-up was 5.5 ± 2.8 years. End points included all-cause mortality and cardiac events, including myocardial infarction and late (>3 months) coronary revascularization. Peak stress wall motion score index was an independent predictor of cardiac events in all age groups (<60 years: hazard ratio [HR] 1.14, P = .02; 60-74 years: HR 1.70, P < .0001; ≥75 years: HR 1.10, P = .006). In patients ≥75 years, peak wall motion score index (HR 1.10, P < .0001) and abnormal left ventricular end-systolic volume response (HR 1.25, P = .03) were independent predictors of death. In patients aged 60 to 74 years, abnormal left ventricular end-systolic volume response (HR 1.43, P = .0003) was independently related to death, whereas in patients <60 years, the echocardiographic data assessed during stress were not a predictor. Dobutamine stress echocardiography provided independent information predictive of cardiac events among all age groups and death in patients ≥60 years. However, among patients <60 years, stress-induced echocardiographic abnormalities were not independently associated with mortality. Comorbidities, which have precluded exercise testing, may be most relevant in predicting mortality in patients <60 years undergoing DSE.


Impact of the COVID-19 Pandemic on the Urologist's clinical practice in Brazil: a management guideline proposal for low- and middle-income countries during the crisis period.
This letter to the Editor aims to provide suggestions and recommendations for the management of urological conditions in times of COVID-19 crisis in Brazil and other low- and middle-income countries. It is important to highlight that one of the main characteristics of this pandemic is the oversaturation of the health system capacity, mostly due to a high demand for personal protective equipment (PPE), Hospital/ICU beds, as well as ventilators. In places with limited resources and where the health care systems are already saturated, such consideration is even more worrisome. Therefore, most worldwide authorities are recommending to avoid, as much as possible, patient's elective visits to hospitals, as well as a judicious use of the operating room in order to mitigate the strain put on the health system. While efforts should be directed to the care of COVID-19 patients, other conditions (especially urgencies and oncological cases) must continue to be assisted. Thus, through a panel of experts, we have prepared a practical guide for Thus, through a panel of experts, we have prepared a practical guide for urologists based on the recommendations from the main Urologic Associations, as well as data from the literature to support the suggested management. We will try to follow the standard guideline recommendations from the American Urological Association (AUA) and European Association of Urology (EAU), with the aim of pursuing the best outcomes possible. However, some recommendations were based on the consensus of the panel, taking into consideration the reality of developing countries and the unprecedented situation caused by the COVID-19 crisis. Most importantly, all recommendations on this manuscript are based on the expectancy of a maximum 3-month duration of the crisis. If this period shall extended, these recommendations will be revised and updated.


Drug-eluting beads transarterial chemoembolization with CalliSpheres microspheres for treatment of unresectable intrahepatic cholangiocarcinoma.
Objective: This study aimed to evaluate the efficacy and safety of doxorubicin-loaded drug-eluting beads transarterial chemoembolization (DEB-TACE) with CalliSpheres microspheres (CSM) in treating unresectable intrahepatic cholangiocarcinoma (ICC). Methods: 88 unresectable ICC patients who received DEB-TACE treatment with CSM were retrospectively enrolled in this study. Information about treatment response, survival and adverse events were collected. The Kaplan-Meier curve was used to evaluate progression-free survival (PFS) and overall survival (OS), and factors affecting OS were determined by Cox's proportional hazards regression model. Results: Tumor response of the whole sample of 88 patients was partial response (PR) in 58 (65.9%) patients, stable disease (SD) in 19 (21.6%) and progressive disease (PD) in 11 (12.5%) at one month after therapy, with no complete responses (CR). The median PFS and OS were 3.0 months and 9.0 months respectively. Cox's proportional hazards regression analysis disclosed that subsequent treatment was an independent favorable prognostic factor, while cholangiectasis, extensive intrahepatic tumor burden and extrahepatic metastasis were the three prognostic factors associated with poor survival in ICC patients. Besides, common adverse events included nausea/vomiting, abdominal pain and transient elevation of liver transaminase in patients treated by DEB-TACE with CSM. Conclusion: DEB-TACE with CSM is safe and well-tolerated for unresectable ICC patients, with a low complication rate and a relative benefit in terms of survival. Subsequent treatments including systemic/loco-regional treatments is an independent favorable prognostic factor, but cholangiectasis, extensive intrahepatic tumor burden and extrahepatic metastases are the three prognostic factors associated with poor survival.


Improving the recognition and diagnosis of fibromyalgia.
Fibromyalgia (FM) is a chronic widespread pain disorder often seen in primary care practices. Advances in the understanding of FM pathophysiology and clinical presentation have improved the recognition and diagnosis of FM in clinical practice. Fibromyalgia is a clinical diagnosis based on signs and symptoms and is appropriate for primary care practitioners to make. The hallmark symptoms used to identify FM are chronic widespread pain, fatigue, and sleep disturbances. Awareness of common mimics of FM and comorbid disorders will increase confidence in establishing a diagnosis of FM.


Remarkably high-Q resonant nanostructures based on atomically thin two-dimensional materials.
Planar optical resonant structures with high quality (Q) factors play a crucial role in modern photonic technologies. In this paper, a type of remarkably high-Q resonant nanostructure based on atomically thin two-dimensional (2D) materials is proposed. It is shown theoretically and numerically that with the excitation of leaky modes in the proposed structures, guided mode resonant (GMR) gratings, can achieve resonances with extremely narrow linewidths down to 0.0005 nm and high Q-factors up to millions in the telecom range. The thickness of 2D materials and thus the high-Q resonances can be precisely controlled by changing the layer number of 2D materials, providing a versatile platform for strong light-matter interactions. As an example, dramatic nonlinear reflectance can be realized around the resonance at a power level of a few kW cm-2 with the Kerr effect. This new type of 2D material resonant nanostructure can be employed for a variety of applications ranging from lasers, filters and polarizers to nonlinear optical devices.


Long term anoxia in rainbow trout investigated by 2-DE and MS/MS.
Twenty-four hours of N(2) induced anoxia induced global perturbations on protein expression in rainbow trout hypodermal fibroblasts cell line. Anoxia was obtained by depleting the medium of O(2) by flushing with N(2), and protein changes were studied by 2-DE coupled with MS providing quantitative measurements of a large number of proteins in one single study. The anoxic insult changed the level of 33 protein spots: 22 of these were up-regulated compared to the control situation and 11 were down-regulated. Using MS/MS sequencing 19 of the 33 protein spots that changed were identified, corresponding to a success rate of more than 50%. The identified proteins included two proteins involved in energy metabolism namely phosphoglycerate mutase and isocitrate dehydrogenase. In addition we observed the up-regulation of a cluster of proteins that contribute to cytoskeleton function. These are calpain, EB1, and Rho GDP dissociation inhibitor (GDI). The up-regulation of Rho GDI was shown to develop in a time dependent manner with no significant increase for up to 8 h of anoxia. In conclusion, this study provides a thorough investigation of the effect of anoxia in a cell line from rainbow trout.


Transcriptional profiling of mechanically and genetically sink-limited soybeans.
The absence of a reproductive sink causes physiological and morphological changes in soybean plants. These include increased accumulation of nitrogen and starch in the leaves and delayed leaf senescence. To identify transcriptional changes that occur in leaves of these sink-limited plants, we used RNAseq to compare gene expression levels in trifoliate leaves from depodded and ms6 male-sterile soybean plants and control plants. In both sink-limited tissues, we observed a deferral of the expression of senescence-associated genes and a continued expression of genes associated with leaf maturity. Gene Ontology-terms (GO-terms) associated with growth and development and storage proteins were over-represented in genes that were differentially expressed in sink-limited tissues. We also identified basic helix-loop-helix, auxin response factor, and squamosa binding protein transcription factors expressed in sink-limited tissues, and the senescing control leaves expressed WRKY and NAC transcription factors. We identified genes that were not expressed during normal leaf development but that were highly expressed in sink-limited plants, including the SGR3b "non-yellowing" gene. These differences highlighted several metabolic pathways that were involved in distinct modes of resource partitioning of leaves with the "stay green" phenotype.


Substrate specificity of the lanthipeptide peptidase ElxP and the oxidoreductase ElxO.
The final step in lanthipeptide biosynthesis involves the proteolytic removal of an N-terminal leader peptide. In the class I lanthipeptide epilancin 15X, this step is performed by the subtilisin-like serine peptidase ElxP. Bioinformatic, kinetic, and mass spectrometric analysis revealed that ElxP recognizes the stretch of amino acids DLNPQS located near the proteolytic cleavage site of its substrate, ElxA. When the ElxP recognition motif was inserted into the noncognate lanthipeptide precursor NisA, ElxP was able to proteolytically remove the leader peptide from NisA. Proteolytic removal of the leader peptide by ElxP during the biosynthesis of epilancin 15X exposes an N-terminal dehydroalanine on the core peptide of ElxA that hydrolyzes to a pyruvyl group. The short-chain dehydrogenase ElxO reduces the pyruvyl group to a lactyl moiety in the final step of epilancin 15X maturation. Using synthetic peptides, we also investigated the substrate specificity of ElxO and determined the 1.85 Å resolution X-ray crystal structure of the enzyme.


Luteinizing hormone receptor mRNA down-regulation is mediated through ERK-dependent induction of RNA binding protein.
The ligand-induced down-regulation of LH receptor (LHR) expression in the ovaries, at least in part, is regulated by a posttranscriptional process mediated by a specific LH receptor mRNA binding protein (LRBP). The LH-mediated signaling pathways involved in this process were examined in primary cultures of human granulosa cells. Treatment with 10 IU human chorionic gonadotropin (hCG) for 12 h resulted in the down-regulation of LHR mRNA expression while producing an increase in LHR mRNA binding to LRBP as well as a 2-fold increase in LRBP levels. The activation of ERK1/2 pathway in LH-mediated LHR mRNA down-regulation was also established by demonstrating the translocation of ERK1/2 from the cytosol to the nucleus using confocal microcopy. Inhibition of protein kinase A using H-89 or ERK1/2 by U0126 abolished the LH-induced LHR mRNA down-regulation. These treatments also abrogated both the increases in LRBP levels as well as the LHR mRNA binding activity. The abolishment of the hCG-induced increase in LRBP levels and LHR mRNA binding activity was further confirmed by transfecting granulosa cells with ERK1/2 specific small interfering RNA. This treatment also reversed the hCG-induced down-regulation of LHR mRNA. These data show that LH-regulated ERK1/2 signaling is required for the LRBP-mediated down-regulation of LHR mRNA.


Association of the DNMT3B -579G>T polymorphism with risk of thymomas in patients with myasthenia gravis.
Increasing evidence suggests a contribution of epigenetic processes in promoting cancer and autoimmunity. Myasthenia gravis (MG) is an autoimmune disease mediated, in approximately 80% of the patients, by antibodies against the nicotinic acetylcholine receptor (AChR+). Moreover, epithelial tumours (thymomas) are present in about 10-20% of the patients, and there is indication that changes in DNA methylation might contribute to the risk and progression of thymomas. However, the role of epigenetics in MG is still not completely clarified. In the present study we investigated if a common polymorphism (-579G>T: rs1569686) in the promoter of the DNMT3B gene coding for the DNA methyltransferase 3B, an enzyme that mediates DNA methylation, increases the risk to develop MG or MG-associated thymomas. The study polymorphism was selected based on recent reports and a literature meta-analysis suggesting association with increased risk of various types of cancer. We screened 324 AChR+ MG patients (140 males and 184 females, mean age 56.0 ± 16.5 years) and 735 healthy matched controls (294 males and 441 females, mean age 57.3 ± 15.6 years). 94 of the total MG patients had a thymoma. While there was no association with the whole cohort of MG patients, we found a statistically significant association of the DNMT3B-579T allele (OR = 1.51; 95% CI=1.1-2.1, P = 0.01) and the TT homozygous genotype (OR = 2.59; 95% CI=1.4-4.9, P = 0.006) with the risk of thymoma. No association was observed in MG patients without thymoma, even after stratification into clinical subtypes. Present results suggest that the DNMT3B-579T allele might contribute to the risk of developing thymoma in MG patients, particularly in homozygous TT subjects.


Regulation of envelope protein composition during adaptation to osmotic stress in Escherichia coli.
Adaptation to osmotic stress alters the amounts of several specific proteins in the Escherichia coli K-12 envelope. The most striking feature of the response to elevated osmolarity was the strong induction of a periplasmic protein with an Mr of 31,000. This protein was absent in mutants with lambda plac Mu insertions in an osmotically inducible locus mapping near 58 min. The insertions are likely to be in proU, a locus encoding a transport activity for the osmoprotectants glycine betaine and proline. Factors affecting the extent of proU induction were identified by direct examination of periplasmic proteins on sodium dodecyl sulfate gels and by measuring beta-galactosidase activity from proU-lac fusions. Expression was stimulated by increasing additions of salt or sucrose to minimal medium, up to a maximum at 0.5 M NaCl. Exogenous glycine betaine acted as an osmoregulatory signal; its addition to the high-osmolarity medium substantially repressed the expression of the 31,000-dalton periplasmic protein and the proU-lac+ fusions. Elevated osmolarity also caused the appearance of a second periplasmic protein (Mr = 16,000), and severe reduction in the amounts of two others. In the outer membrane, the well-characterized repression of OmpF by high osmolarity was observed and was reversed by glycine betaine. Additional changes in membrane composition were also responsive to glycine betaine regulation.


[The bilateral interaction of the hypothalamus, amygdala and neocortex at different stages in conditioned reflex acquisition].
By averaged evoked potentials of the symmetric points of the cat's auditory cortex (Al), hypothalamus (Hl) and amygdala (Abl) in response to a sound was determined the character of the two-way interaction (Spearman coefficient of correlation) between the structures before and at different stages of instrumental food-procuring conditioning. Individual character of co-tuning of Al, Hl, and Abl as well as the emotional state connected with adaptation of an animal were found to be reflected in the correlation of evoked activity of the structures. Conditioning led to a formation of close interconnections between the structures eliminating individual peculiarities of correlation. Stabilization of the response was accompanied by a weakening of the interconnections between the cortex and the subcortical structures, internal symmetrical co-tuning of Al, Hl and Abl being unchanged.


Immunophenotypic study of basophils by multiparameter flow cytometry.
The immunophenotypic profile of basophils is not yet fully established, and the immunophenotypic changes in chronic myelogenous leukemia are not fully characterized. To establish a comprehensive immunophenotypic spectrum of normal basophils and to assess the range of immunophenotypic aberrations of basophils in chronic myelogenous leukemia. Using 4-color flow cytometry, we compared the immunophenotypic profile of basophils in peripheral blood or bone marrow samples from 20 patients with no evidence of neoplasia to basophils from 15 patients with chronic myelogenous leukemia. Basophils in control cases were all positive for CD9, CD13, CD22, CD25 (dim), CD33, CD36, CD38 (bright), CD45 (dimmer than lymphocytes and brighter than myeloblasts), and CD123 (bright), and were negative for CD19, CD34, CD64, CD117, and HLA-DR. Basophils in all chronic myelogenous leukemia patients possessed 1 to 5 immunophenotypic aberrancies. The most common aberrancies were underexpression of CD38, followed by aberrant expression of CD64 and underexpression of CD123. CD34 and CD117 were present in cases with basophilic precursors. Myeloblasts showed a distinct immunophenotypic profile, as they typically expressed CD34 and CD117, showed dimmer expression (compared with basophils) of CD38, CD45, and CD123, and lacked expression of CD22. Flow cytometric immunophenotyping can identify immunophenotypic aberrations of basophils in chronic myelogenous leukemia, and discriminate basophils from myeloblasts.


Genetic polymorphisms as prognostic factors for recurrent kidney stones: A systematic review and meta-analysis.
Genetic polymorphisms have been suggested as risk factors affecting the occurrence and recurrence of kidney stones, although findings regarding the latter remain inconclusive. We performed this systematic review and meta-analysis to clarify the associations between genetic polymorphisms and recurrent kidney stones. PubMed, SCOPUS, EMBASE, and Cochrane Library databases were searched through May 28th, 2020 to identify eligible studies. The Quality in prognostic studies (QUIPS) tool was used to evaluate bias risk. Allelic frequencies and different inheritance models were assessed. All analyses were performed using Review manager 5.4. A total of 14 studies were included for meta-analysis, assessing urokinase (ApaL1) and vitamin D receptor (VDR) (ApaI, BsmI, FokI, and TaqI) gene polymorphisms. The ApaLI polymorphism demonstrated protective association in the recessive model [odds ratio (OR) 0.45, P < 0.01] albeit higher risk among Caucasians in the heterozygous model (OR 16.03, P < 0.01). The VDR-ApaI polymorphism showed protective association in the dominant model (OR 0.60, P < 0.01). Among Asians, the VDR-FokI polymorphism recessive model showed significant positive association (OR 1.70, P < 0.01) and the VDR-TaqI polymorphism heterozygous model exhibited protective association (OR 0.72, P < 0.01). The VDR-BsmI polymorphism was not significantly associated with recurrent kidney stones in any model. Urokinase-ApaLI (recessive model), VDR-ApaI (dominant model), and VDR-TaqI (heterozygous model) polymorphisms were associated with decreased recurrent kidney stone risk whereas urokinase-ApaLI (heterozygous model) and VDR-FokI polymorphisms were associated with increased risk among Caucasians and Asians, respectively. These findings will assist in identifying individuals at risk of kidney stone recurrence.


Evaluating the impact of a single-day multidisciplinary clinic on the management of pancreatic cancer.
To evaluate the impact of a multidisciplinary clinic on the clinical care recommendations of patients with pancreatic cancer compared with the recommendations the patients received prior to review by the multidisciplinary tumor board. The records of 203 consecutive patients referred to the Johns Hopkins pancreatic multidisciplinary clinic were prospectively collected from November 2006 to October 2007. Cross-sectional imaging, pathology, and medical history were evaluated by a panel of medical/radiation oncologists, surgical oncologists, pathologists, diagnostic radiologists, and geneticists. Alterations in treatment recommendations between the outside institution and the multidisciplinary clinic were recorded and compared. On presentation, the outside computed tomography (CT) report described locally advanced/unresectable disease (34.9%), metastatic disease (17.7%), and locally advanced disease with metastasis (1.1%). On review of submitted imaging and imaging performed at Hopkins, 38 out of 203 (18.7%) patients had a change in the status of their clinical stage. Review of the histological slides by dedicated pancreatic pathologists resulted in changes in the interpretation for 7 of 203 patients (3.4%). Overall, 48 out of 203 (23.6%) patients had a change in their recommended management based on clinical review of their case by the multidisciplinary tumor board. Enrollment into the National Familial Pancreas Tumor Registry increased from 52 out of 106 (49.2%) patients in 2005 to 158 out of 203 (77.8%) with initiation of the multidisciplinary clinic. The single-day pancreatic multidisciplinary clinic provided a comprehensive and coordinated evaluation of patients that led to changes in therapeutic recommendations in close to one-quarter of patients.


Unprecedented CO2-promoted hydrogen permeation in Ni-BaZr0.1Ce0.7Y0.1Yb0.1O(3-δ) membrane.
Conventional Ni-BaCeO3-based membranes possess high hydrogen permeation flux but suffer serious flux degradation in CO2-containing atmosphere because of the formation of BaCO3 insulating layer. In this work, we report a novel Ni-BaZr0.1Ce0.7Y0.1Yb0.1O(3-δ) (Ni-BZCYYb) membrane, capable of both high hydrogen permeation flux and stable performance in CO2-containing atmosphere at 900 °C. Most importantly, the flux is found to be promoted rather than being diminished by CO2 normally observed for other high temperature proton conductors. The flux enhancement in Ni-BZCYYb membrane is attributed to the increase of moisture content in feed gas. When CO2 is introduced, the reverse water-gas shift reaction takes place generating H2O and CO. This work demonstrates that CO2 can be beneficial rather than detrimental for hydrogen permeation membranes that possess high chemical stability.


Comparative study between three analgesic agents for the pain management during extracorporeal shock wave lithotripsy.
To compare the clinical efficacy between locally applied diclofenac diethylamine gel, EMLA cream and systemically given diclofenac sodium for the pain relief during extracorporeal shock wave lithotripsy (SWL) using Dornier Delta Compact Lithotripter. One hundred five patients with renal stones were randomly divided in to 3 groups. Group A was given intramuscular diclofenac sodium (1 mg/kg), 45 minutes before the procedure. In group B, 10 gm of eutectic mixture of local anesthetic (EMLA) cream and in group C, 15 gm of diclofenac diethylamine gel was applied locally 45 minutes before the procedure. Ten-score linear and visual analogue scale (VAS) was used to assess the severity of pain during the procedure. Analysis of variance (ANOVA) test was used to compare various parameters and analyzed statistically. All the three groups were not statistically different with respect to age, weight, stone size, number of shock wave delivered and maximum voltage used (P > .05). The mean pain score in group A was 4.48, in group B was 3.60 and in group C was 3.95, which were not significantly different (P = 1.34). Complication like skin lesion was found only in injection diclofenac sodium group whereas cold sensation at the local site was typically found in diclofenac diethylamine gel group. Although not statistically significant, the mean pain score in locally applied analgesic agents (EMLA and diclofenac diethylamine gel) is lower as compared to intramuscularly given diclofenac sodium. Among these two locally acting drugs, diclofenac diethylamine gel is an equally effective alternative to EMLA.


Longitudinal development of central SBP in children with congenital heart disease.
Central SBP (cSBP) was shown to be increased already in children with congenital heart disease (CHD). However, its development over time has not yet been investigated. The aim of this study was to evaluate the natural course of cSBP over time from longitudinal assessment in children with CHD. In this longitudinal study, 306 children and adolescents (11.3 ± 2.9 years, 34% girls) with various CHD were prospectively examined from July 2014 to May 2022. Over a mean follow-up length of 30.1 ± 18.9 months, 467 follow-up assessments have been conducted. cSBP was noninvasively assessed by oscillometric measurement via Mobil-O-Graph. A linear mixed effect model was performed to examine the course of cSBP. cSBP increased significantly over time by 1.22 mmHg per year of age (P < 0.001). The longitudinal increase in cSBP over time remained significant when including sex (b = 0.68, P < 0.001), BMI (b = 1.12, P < 0.001), hypertensive medication (b = 1.13, P < 0.001), disease severity (b = 1.04, P < 0.001), and CHD type (b = 3.74, P = 0.03) in the model. Patients with transposition of the great arteries (TGA) after arterial switch had a significantly higher cSBP increase over time (b = 1.78, P < 0.001). The longitudinal cSBP increase was significantly higher in obese CHD children (b = 2.52, P = 0.005) and in boys (b = 0.85, P < 0.001). This study shows a longitudinal increase in cSBP in children with CHD. Whether observed trajectories of cSBP are normal or abnormal needs to be investigated in further studies. Monitoring of the vascular function with a special focus on patients with TGA and obese CHD children seems indicated.


Preparation and characterization of metformin hydrochloride loaded-Eudragit®RSPO and Eudragit®RSPO/PLGA nanoparticles.
The aim of the present study was to develop and characterize metformin HCl-loaded nanoparticle formulations. Nanoparticles were prepared by the nanoprecipitation method using both a single polymer (Eudragit(®)RSPO) and a polymer mixture (Eudragit/PLGA). The mean particle size ranged from 268.8 to 288 nm and the nanoparticle surface was positively charged (9.72 to 10.1 mV). The highest encapsulation efficiency was observed when Eudragit®RSPO was used. All formulations showed highly reproducible drug release profiles and the in vitro drug release in phosphate buffer (pH = 6.8) ranged from 92 to 100% in 12 h. These results suggest that Eudragit(®)RSPO or Eudragit/PLGA nanoparticles might represent a promising sustained-release oral formulation for metformin HCl, reducing the necessity of repeated administrations of high doses to maintain effective plasma concentrations, and thus, increasing patient compliance and reducing the incidence of side-effects.


ITIH5 inhibits proliferation, adipogenic differentiation, and secretion of inflammatory cytokines of human adipose stem cells-A new key in treating obesity?
Inter-α-trypsin inhibitor heavy chain 5 (ITIH5) is widely expressed in the human body, and it is detected to be particularly abundant in adipose tissue. ITIH5 expression is increased in people with obesity compared to lean persons and is decreased by diet-induced weight loss. This suggests that ITIH5 may be involved in the development of adiposity and clinical metabolic variables, although its exact function remains unknown. We measured the protein concentration of ITIH5 in adipose samples from patients undergoing abdominoplasty and tested for correlation with the subjects' BMI as well as inflammatory mediators. We stimulated human adipose stem cells (ASCs) with recombinant (r)ITIH5 protein and tested for an effect on proliferation, differentiation, and immunosuppressive properties when the cells were exposed to an artificial inflammatory environment. We found positive correlations between ITIH5 levels and the BMI (p < .001) as well as concentrations of inflammatory cytokines (TNF-α, IL-6, and MCP-1) in adipose tissue (p < .01). Application of the rITIH5 protein inhibited both proliferation (p < .001) and differentiation of ASCs. Especially, the development of mature adipocytes was reduced by over 50%. Moreover, rITIH5 decreased the release of IL-6 and MCP-1 when the cells were exposed to TNF-α and IL-1β (p < .001). Our data suggest that ITIH5 is an adipokine that is increasingly released during human adipose tissue development, acting as a regulator that inhibits proliferation and adipogenic differentiation of ASCs. ITIH5 thus presents itself as a positive regulator of adipose tissue homeostasis, possibly protecting against both hyperplasia and hypertrophy of adipose tissue and the associated chronic inflammation.


Spanning high-dimensional expression space using ribosome-binding site combinatorics.
Protein levels are a dominant factor shaping natural and synthetic biological systems. Although proper functioning of metabolic pathways relies on precise control of enzyme levels, the experimental ability to balance the levels of many genes in parallel is a major outstanding challenge. Here, we introduce a rapid and modular method to span the expression space of several proteins in parallel. By combinatorially pairing genes with a compact set of ribosome-binding sites, we modulate protein abundance by several orders of magnitude. We demonstrate our strategy by using a synthetic operon containing fluorescent proteins to span a 3D color space. Using the same approach, we modulate a recombinant carotenoid biosynthesis pathway in Escherichia coli to reveal a diversity of phenotypes, each characterized by a distinct carotenoid accumulation profile. In a single combinatorial assembly, we achieve a yield of the industrially valuable compound astaxanthin 4-fold higher than previously reported. The methodology presented here provides an efficient tool for exploring a high-dimensional expression space to locate desirable phenotypes.


[Immune reactivity in workers engaged in cement production].
The authors studied immune state of workers engaged in cement production. Ambivalent changes of immune parameters in apparently healthy workers depict various stages of adaptational protective reactions in body system. The studies support gradual changes in immune system under continued influence of occupational factors.


Regulation of pyruvate dehydrogenase (PDH) in the hibernating ground squirrel, (Ictidomys tridecemlineatus).
Pyruvate dehydrogenase (PDH) is a vital regulatory enzyme that catalyzes the conversion of pyruvate into acetyl-CoA and connects anaerobic glycolysis to aerobic TCA cycle. Post-translational inhibition of PDH activity via three serine phosphorylation sites (pS232, pS293, and pS300) regulate the metabolic flux through the TCA cycle, decrease glucose utilization, and facilitate lipid metabolism during times of nutrient deprivation. As metabolic readjustment is necessary to survive hibernation, the purpose of this study was to explore the post-translational regulation of pyruvate dehydrogenase and the expression levels of four mitochondrial serine/threonine kinases (PDHKs), during torpor-arousal cycles in liver, heart, and skeletal muscle of 13-lined ground squirrels. A combination of Luminex multiplex technology and western immunoblotting were used to measure the protein expression levels of total PDH, three phosphorylation sites, S232, 293, 300, and the expression levels of the corresponding PDH kinases (PDHK1-4) during euthermic control, entrance, late torpor, and interbout arousal. Liver and heart showed strong inhibitory PDH regulation, indicating a possible decrease in glucose utilization and a possible preference for β-oxidation of fatty acids during periods of low temperature and starvation. On the contrary, skeletal muscle showed limited PDH regulation via phosphorylation, possibly due to alternate controls. Phosphorylation of PDH may play an important role in regulating aerobic and anaerobic metabolic responses during hibernation in the 13-lined ground squirrel.


NanoLC-MS/MS analysis provides new insights into the phosphorylation pattern of Cdc25B in vivo: full overlap with sites of phosphorylation by Chk1 and Cdk1/cycB kinases in vitro.
NanoLC-MS/MS analysis was used to characterize the phosphorylation pattern in vivo of CDC25B3 (phosphatase splice variant 1) expressed in a human cell line and to compare it to the phosphorylation of CDC25B3 by Cdk1/cyclin B and Chk1 in vitro. Cellular CDC25B3 was purified from U2OS cells conditionally overexpressing the phosphatase. Eighteen sites were detectably phosphorylated in vivo. Nearly all existing (S/T)P sites were phosphorylated in vivo and in vitro. Eight non(S/T)P sites were phosphorylated in vivo. All these sites could be phosphorylated by kinase Chk1, which phosphorylated a total of 11 sites in vitro, with consensus sequence (R/K) X(2-3) (S/P)-non P. Nearly half of the sites identified in this study were not previously described and were not homologous to sites reported to be phosphorylated in other CDC25 species. We also show that in vivo a significant part of CDC25B molecules can be hyperphosphorylated, with up to 13 phosphates per phosphatase molecule.


vttRA and vttRB Encode ToxR family proteins that mediate bile-induced expression of type three secretion system genes in a non-O1/non-O139 Vibrio cholerae strain.
Strain AM-19226 is a pathogenic non-O1/non-O139 serogroup Vibrio cholerae strain that does not encode the toxin-coregulated pilus or cholera toxin but instead causes disease using a type three secretion system (T3SS). Two genes within the T3SS pathogenicity island, herein named vttR(A) (locus tag A33_1664) and vttR(B) (locus tag A33_1675), are predicted to encode proteins that show similarity to the transcriptional regulator ToxR, which is found in all strains of V. cholerae. Strains with a deletion of vttR(A) or vttR(B) showed attenuated colonization in vivo, indicating that the T3SS-encoded regulatory proteins play a role in virulence. lacZ transcriptional reporter fusions to intergenic regions upstream of genes encoding the T3SS structural components identified growth in the presence of bile as a condition that modulates gene expression. Under this condition, VttR(A) and VttR(B) were necessary for maximal gene expression. In contrast, growth in bile did not substantially alter the expression of a reporter fusion to the vopF gene, which encodes an effector protein. Increased vttR(B) reporter fusion activity was observed in a DeltavttR(B) strain background, suggesting that VttR(B) may regulate its own expression. The collective results are consistent with the hypothesis that T3SS-encoded regulatory proteins are essential for pathogenesis and control the expression of selected T3SS genes.


Training phonological awareness with and without explicit attention to articulation.
One hundred twenty-two second- to fifth-grade (7- to 11-year-old) children with reading difficulties studied phonological awareness with or without explicit attention to articulation and with or without manipulation of sounds. They all studied identical phonics and read stories on the computer with speech and decoding support for difficult words. Regular-instruction controls received regularly scheduled language-arts or reading activities. After 40 h of training, children in all three trained conditions outperformed controls on all tests except math. Conditions that manipulated sounds showed advantages over the condition without explicit practice manipulating sounds, but only on the two measures of phonological awareness. Articulatory awareness training yielded no unique benefits during this training period. Individual differences in response to treatment related to initial levels of phonological awareness, naming speed, IQ, and grade. The similar outcomes of the three conditions suggest that specific variations in good phonological training may be less important than once thought for most children with reading difficulties.


The association of angiotensin-converting enzyme gene insertion/deletion polymorphisms with OSA: a meta-analysis.
Obstructive sleep apnoea (OSA) is an independent risk factor for hypertension. Increased angiotensin-converting enzyme (ACE) activity may be a possible promoting mechanism with different ACE insertion/deletion (I/D) genotypes influencing this activity. Studies investigating the association of ACE I/D polymorphisms with OSA have shown conflicting results. We aimed to undertake a meta-analysis of existing studies exploring the association of ACE I/D polymorphisms with the risk of OSA and hypertension. 10 studies were included in a random effects meta-analysis, comprising 1,227 OSA subjects and 1,227 controls. The effect size was measured using the odds ratio. The risk of having OSA in carriers of the D allele was 0.92 (95% CI 0.69-1.23). There was statistically significant heterogeneity across the studies (I(2)=42%, p=0.08 and I(2)=74%, p<0.0001 for genotype and allele frequency, respectively). The association of D allele frequency with the risk of OSA remained nonsignificant after stratification based on ethnicity, source of population sample, and the presence of hypertension. Subgroup analysis failed to show any influence of genotype and allele frequency on OSA severity. This meta-analysis revealed no association between the ACE I/D polymorphisms and OSA susceptibility.


M-side electron transfer in reaction center mutants with a lysine near the nonphotoactive bacteriochlorophyll.
We report the primary charge separation events in a series of Rhodobacter capsulatus reaction centers (RCs) that have been genetically modified to contain a lysine near the bacteriochlorophyll molecule, BChl(M), on the nonphotoactive M-side of the RC. Using wild type and previously constructed mutants as templates, we substituted Lys for the native Ser residue at position 178 on the L polypeptide to make the S(L178)K single mutant, the S(L178)K/G(M201)D and S(L178)K/L(M212)H double mutants, and the S(L178)K/G(M201)D/L(M212)H triple mutant. In the triple mutant, the decay of the photoexcited primary electron donor (P) occurs with a time constant of 15 ps and is accompanied by 15% return to the ground state, 62% electron transfer to the L-side bacteriopheophytin, BPh(L), and 23% electron transfer to the M-side analogue, BPh(M). The data supporting electron transfer to the M-side include bleaching of the Q(X) band of BPh(M) at 528 nm and a spectrally and kinetically resolved anion band with a maximum at 640 nm assigned to BPh(M)(-). The decay of these features and concomitant approximately 20% decay of bleaching of the 850 nm band of P give a P(+)BPh(M)(-) lifetime on the order of 1-2 ns that reflects deactivation to give the ground state. These data and additional findings are compared to those from parallel experiments on the G(M201)D/L(M212)H double mutant, in which 15% electron transfer to BPh(M) has been reported previously and is reproduced here. We also compare the above results with the primary electron-transfer processes in S(L178)K, S(L178)K/G(M201)D, and S(L178)K /L(M212)H RCs and with those for the L(M212)H and G(M201)D single mutants and wild-type RCs. The comparison of extensive results that track the primary events in these eight RCs helps to elucidate key factors underlying the directionality and high yield of charge separation in the bacterial photosynthetic RC.


High dynamic range head mounted display based on dual-layer spatial modulation.
The human visual system can perceive a much wider range of contrast than the addressable dynamic range afforded by the state-of-art head mounted displays (HMD). Increasing the dynamic range of HMDs becomes critical especially for augmented reality applications where the dynamic range of outdoor scenes can be as large as 14 orders of magnitude. In this paper, we present the integrated work of the design, implementation, calibration, and image-rendering algorithm of a high dynamic range HMD system. By using a pair of LCoS microdisplays as the spatial light modulators, accompanied with the relay optics to optically overlay the modulation layers, we demonstrate the reconstruction of high dynamic range images with high accuracy.


Isoprene emission rates under elevated CO2 and O3 in two field-grown aspen clones differing in their sensitivity to O3.
Isoprene is the most important nonmethane hydrocarbon emitted by plants. The role of isoprene in the plant is not entirely understood but there is evidence that it might have a protective role against different oxidative stresses originating from heat shock and/or exposure to ozone (O(3)). Thus, plants under stress conditions might benefit by constitutively high or by higher stress-induced isoprene emission rates. In this study, measurements are presented of isoprene emission from aspen (Populus tremuloides) trees grown in the field for several years under elevated CO(2) and O(3). Two aspen clones were investigated: the O(3)-tolerant 271 and the O(3)-sensitive 42E. Isoprene emission decreased significantly both under elevated CO(2) and under elevated O(3) in the O(3)-sensitive clone, but only slightly in the O(3)-tolerant clone. This study demonstrates that long-term-adapted plants are not able to respond to O(3) stress by increasing their isoprene emission rates. However, O(3)-tolerant clones have the capacity to maintain higher amounts of isoprene emission. It is suggested that tolerance to O(3) is explained by a combination of different factors; while the reduction of O(3) uptake is likely to be the most important, the capacity to maintain higher amounts of isoprene is an important factor in strengthening this character.


Formulation design considerations for oral vaccines.
Whilst oral vaccination is a potentially preferred route in terms of patient adherence and mass vaccination, the ability to formulate effective oral vaccines remains a challenge. The primary barrier to oral vaccination is effective delivery of the vaccine through the GI tract owing to the many obstacles it presents, including low pH, enzyme degradation and bile-salt solubilization, which can result in breakdown/deactivation of a vaccine. For effective immune responses after oral administration, particulates need to be taken up bythe M cells however, these are few in number. To enhance M-cell uptake, particle characteristics can be optimized with particle size, surface charge, targeting groups and bioadhesive properties all being considerations. Yet improved uptake may not translate into enhanced immune responses and formulating particulates with inherent adjuvant properties can offer advantages. Within this article, we establish the options available for consideration when building effective oral particulate vaccines.


Regulation of E2F1-induced apoptosis by poly(ADP-ribosyl)ation.
The transcription factor adenovirus E2 promoter-binding factor (E2F)-1 normally enhances cell-cycle progression, but it also induces apoptosis under certain conditions, including DNA damage and serum deprivation. Although DNA damage facilitates the phosphorylation and stabilization of E2F1 to trigger apoptosis, how serum starvation renders cells vulnerable to E2F1-induced apoptosis remains unclear. Because poly(ADP-ribose) polymerase 1 (PARP1), a nuclear enzyme essential for genomic stability and chromatin remodeling, interacts directly with E2F1, we investigated the effects of PARP1 on E2F1-mediated functions in the presence and absence of serum. PARP1 attenuation, which increased E2F1 transactivation, induced G2/M cell-cycle arrest under normal growth conditions, but enhanced E2F1-induced apoptosis in serum-starved cells. Interestingly, basal PARP1 activity was sufficient to modify E2F1 by poly(ADP-ribosyl)ation, which stabilized the interaction between E2F1 and the BIN1 tumor suppressor in the nucleus. Accordingly, BIN1 acted as an RB1-independent E2F1 corepressor. Because E2F1 directly activates the BIN1 gene promoter, BIN1 curbed E2F1 activity through a negative-feedback mechanism. Conversely, when the BIN1-E2F1 interaction was abolished by PARP1 suppression, E2F1 continuously increased BIN1 levels. This is functionally germane, as PARP1-depletion-associated G2/M arrest was reversed by the transfection of BIN1 siRNA. Moreover, PARP-inhibitor-associated anti-transformation activity was compromised by the coexpression of dominant-negative BIN1. Because serum starvation massively reduced the E2F1 poly(ADP-ribosyl)ation, we conclude that the release of BIN1 from hypo-poly(ADP-ribosyl)ated E2F1 is a mechanism by which serum starvation promotes E2F1-induced apoptosis.


Expression of tryptophan 2,3-dioxygenase in mature granule cells of the adult mouse dentate gyrus.
New granule cells are continuously generated in the dentate gyrus of the adult hippocampus. During granule cell maturation, the mechanisms that differentiate new cells not only describe the degree of cell differentiation, but also crucially regulate the progression of cell differentiation. Here, we describe a gene, tryptophan 2,3-dioxygenase (TDO), whose expression distinguishes stem cells from more differentiated cells among the granule cells of the adult mouse dentate gyrus. The use of markers for proliferation, neural progenitors, and immature and mature granule cells indicated that TDO was expressed in mature cells and in some immature cells. In mice heterozygous for the alpha-isoform of calcium/calmodulin-dependent protein kinase II, in which dentate gyrus granule cells fail to mature normally, TDO immunoreactivity was substantially downregulated in the dentate gyrus granule cells. Moreover, a 5-bromo-2'-deoxyuridine labeling experiment revealed that new neurons began to express TDO between 2 and 4 wk after the neurons were generated, when the axons and dendrites of the granule cells developed and synaptogenesis occurred. These findings indicate that TDO might be required at a late-stage of granule cell development, such as during axonal and dendritic growth, synaptogenesis and its maturation.


NF-kappa B signature on the aging wall.
During the lifetime, our cells encounter many challenges to their own existence. The survival of organisms and cells depends on the interplay within a complex, yet precisely orchestrated network of proteins. Since the discovery of the Rel/NF-kappaB family of transcription factors, the research has never come to a stop. These proteins are best known for their key roles played in normal immune and inflammatory responses, also in the control of cellular proliferation, differentiation, apoptosis and oncogenesis. The recent years have witnessed an emerging research regarding NF-kappaB signalling pathways in aging and senescence as well. In this review we go deeply through NF-kappaB activity in the modern biogerontology, with the arguments around its possible roles in aging. Nevertheless, a detailed molecular action and mechanism of NF-kappaB regulation during senescence has yet to be determined. Elucidation of the exact role of NF-kappaB in aging will be crucial for developing new therapeutic strategies.


Novel drugs and current therapeutic approaches in the treatment of heart failure.
Treatment of heart failure attempts to reduce symptoms, increase functional capacity and prolong survival. Optimal therapy usually requires a combination of several drugs. At present, ACE inhibitors are the drugs of first choice, but must be combined with diuretics in symptomatic patients. Digitalis glycosides are still an important supplement to diuretics and ACE inhibitors. Specific angiotensin receptor antagonists such as losartan have an effect comparable with that of ACE inhibitors and may possess certain advantages because of their direct effect at the receptor level. Extensive research has been conducted in the treatment of heart failure. Newer direct acting vasodilators such as flosequinan and epoprostenol have demonstrated improved exercise tolerance but have an adverse effect on mortality. Positive inotropic agents consisting of a heterogeneous group of drugs have been evaluated. Although novel agents such as xamoterol, milrinone, pimobendan and vesnarinone have demonstrated improved haemodynamics and improved symptoms, they are not advisable at present due to increased mortality related to treatment or a high incidence of adverse events. beta-Blockers, used judiciously, may improve functional capacity as well as mortality and may be an important supplement to current conventional treatment. The new generation of beta-blockers with vasodilating properties such as carvedilol and bucindolol appear promising.


Mesenchymal stem cells reduce cigarette smoke-induced inflammation and airflow obstruction in rats via TGF-β1 signaling.
Cigarette smoke has been shown to cause chronic inflammation of the lungs, eventually leading to chronic obstructive pulmonary disease (COPD). Additionally, recent studies have suggested that mesenchymal stem cells (MSCs) can mediate local inflammatory responses in the lungs. Thus, the aim of the present study was to test the effects of rat MSCs (rMSCs) on inflammation of the lungs and destructive pulmonary function induced by cigarette smoke in rats. Rats were exposed to cigarette smoke for 7 weeks. rMSCs were cultured in vitro and infused intratracheally into cigarette smoke-exposed rats. The total and differential cell counts in the bronchoalveolar lavage fluid (BALF), histological changes, pro-inflammatory cytokines, transforming growth factor-β1 (TGF-β1) expression, and pulmonary function were evaluated. Additionally, human peripheral blood mononuclear cells and human MSCs were cocultured in vitro to detect cytokines and TGF-β1 levels. We found that rMSC administration resulted in downregulation of pro-inflammatory cytokines in the lungs while increasing TGF-β1 expression, reducing total inflammatory cell numbers in the BALF, and improving pulmonary histopathology and airflow obstruction. Coculture revealed that human MSCs mediated an anti-inflammatory effect partly via upregulation of TGF-β1. These findings suggested that MSCs may have therapeutic potential in cigarette smoke-induced inflammation and airflow obstruction, partly via upregulation of TGF-β1.


Increased frequency of thyroid nodules and breast fibroadenomas in women with uterine fibroids.
Fibroids or leiomyomas or myomas of the uterus are the most common benign gynecologic disease, while fibroadenomas of the breast are most frequently seen in young women, usually within 20 years after puberty. Multiple tumors in one or both breasts are found in 10-15% of patients. Single thyroid nodules are much more common in women than in men, and their prevalence increases with age. The aim of the present study was to determine cross-sectionally the incidence of solitary thyroid nodules and fibroadenomas of the breast in women with uterine fibroids. In women with uterine adenomas, the frequency of fibroadenomas of the breast was 65% and of thyroid nodules was 38.7%, while in women with a normal uterus, the frequency was 35% and 20%, respectively. Therefore, women with uterine fibroids have an increased incidence of thyroid nodules (t = 4.68, p = 0.030) and of fibroadenomas of the breast (t = 11.74, p = 0.001).


Global patterns and trends in human-wildlife conflict compensation.
Human-wildlife conflict is a major conservation challenge, and compensation for wildlife damage is a widely used economic tool to mitigate this conflict. The effectiveness of this management tool is widely debated. The relative importance of factors associated with compensation success is unclear, and little is known about global geographic or taxonomic differences in the application of compensation programs. We reviewed research on wildlife-damage compensation to determine geographic and taxonomic gaps, analyze patterns of positive and negative comments related to compensation, and assess the relative magnitude of global compensation payments. We analyzed 288 publications referencing wildlife compensation and identified 138 unique compensation programs. These publications reported US$222 million (adjusted for inflation) spent on compensation in 50 countries since 1980. Europeans published the most articles, and compensation funding was highest in Europe, where depredation by wolves and bears was the most frequently compensated damage. Authors of the publications we reviewed made twice as many negative comments as positive comments about compensation. Three-quarters of the negative comments related to program administration. Conversely, three-quarters of the positive comments related to program outcomes. The 3 most common suggestions to improve compensation programs included requiring claimants to employ damage-prevention practices, such as improving livestock husbandry or fencing of crops to receive compensation (n = 25, 15%); modifying ex post compensation schemes to some form of outcome-based performance payment (n = 21, 12%); and altering programs to make compensation payments more quickly (n = 14, 8%). We suggest that further understanding of the strengths and weaknesses of compensation as a conflict-mitigation tool will require more systematic evaluation of the factors driving these opinions and that differentiating process and outcomes and understanding linkages between them will result in more fruitful analyses and ultimately more effective conflict mitigation.


Croconaine-based nanoparticles enable efficient optoacoustic imaging of murine brain tumors.
Contrast enhancement in optoacoustic (photoacoustic) imaging can be achieved with agents that exhibit high absorption cross-sections, high photostability, low quantum yield, low toxicity, and preferential bio-distribution and clearance profiles. Based on advantageous photophysical properties of croconaine dyes, we explored croconaine-based nanoparticles (CR780RGD-NPs) as highly efficient contrast agents for targeted optoacoustic imaging of challenging preclinical tumor targets. Initial characterization of the CR780 dye was followed by modifications using polyethylene glycol and the cancer-targeting c(RGDyC) peptide, resulting in self-assembled ultrasmall particles with long circulation time and active tumor targeting. Preferential bio-distribution was demonstrated in orthotopic mouse brain tumor models by multispectral optoacoustic tomography (MSOT) imaging and histological analysis. Our findings showcase particle accumulation in brain tumors with sustainable strong optoacoustic signals and minimal toxic side effects. This work points to CR780RGD-NPs as a promising optoacoustic contrast agent for potential use in the diagnosis and image-guided resection of brain tumors.


Quantitative assessment of polymerization-binding mechanics of cyanoacrylates: model development and validation.
Although commonly acknowledged as paramount in significance, the mechanics of cyanoacrylate polymerization remain poorly characterized and quantified for clinical applications. This prompted development of a simplistic model for the systematic study of polymerization and binding behaviors of cyanoacrylates. A sliding bed apparatus was constructed that linked a strain gage with a vessel that could be filled with liquid medium, cyanoacrylate, and a microcatheter. As the cyanoacrylate polymerized, the microcatheter was mechanically drawn away very slowly from the fixed vessel, resulting in the development of forces that were recorded to characterize the dynamics of polymerization and binding. Optimization of the model required manipulation of several variables that could influence polymerization. Three different formulations of cyanoacrylate were also tested to determine whether there are significant differences in polymerization dynamics. After experimenting with a few basic physical parameters of the test apparatus, consistent measurements of binding forces during cyanoacrylate polymerization could be recorded and measured. Polymerization produced a multiphasic pattern of binding forces, in which three distinct phases were observed. Furthermore, the rates of polymerization were significantly influenced by a variety of parameters, including the type of fluid within the model vessel, geometry of the model vessel, and rate of injection of acrylic into the well. Furthermore, there were significant differences in the pattern of dynamic binding forces among the various formulations of cyanoacrylate tested. A standardized bench top testing apparatus has been developed, which can consistently show dynamic binding related to polymerization of cyanoacrylates. This preliminary study shows a clear multiphasic pattern of polymerization binding, which may have important clinical implications. The apparatus may be useful for gaining better insight into a variety of clinically important phenomena related to cyanoacrylate polymerization.


Impact of Reducing Time-to-Antibiotics on Sepsis Mortality, Antibiotic Use, and Adverse Events.
Shorter time-to-antibiotics is lifesaving in sepsis, but programs to hasten antibiotic delivery may increase unnecessary antibiotic use and adverse events. We sought to estimate both benefits and harms of shortening time-to-antibiotics for sepsis. Simulation study using a cohort of 1,559,523 hospitalizations via emergency department with ≥2 systemic inflammatory response syndrome criteria (2013-2018). Hospitalizations were classified as septic shock, sepsis, infection, antibiotics stopped early, and never treated (no antibiotics within 48 hours). We simulated the impact of a 50% reduction in time-to-antibiotics for sepsis across 12 hospital scenarios defined by sepsis prevalence (low, medium, high) and magnitude of "spillover" antibiotic prescribing to patients without infection (low, medium, high, very high). Outcomes included mortality and adverse events potentially attributable to antibiotics (allergy, organ dysfunction, Clostridiodes difficile infection, and culture with multi-drug resistant organism [MDRO]). 933,458 (59.9%) hospitalizations received antimicrobial therapy within 48 hours of presentation, including 38,572 (2.5%) with septic shock, 276,082 (17.7%) with sepsis, 370,705 (23.8%) with infection, and 248,099 (15.9%) with antibiotics stopped early. 199,937 (12.8%) hospitalizations experienced an adverse event, most commonly acute liver injury (5.6%), new MDRO (3.5%), and Clostridiodes difficile infection (1.7%). Across the scenarios, a 50% reduction in time-to-antibiotics for sepsis was associated with a median 1 to 180 additional antibiotic-treated patients and 0 to 7 additional adverse events per death averted from sepsis. The impacts of faster time-to-antibiotics for sepsis vary markedly across simulated hospital types. However, even in the worst-case scenario, new antibiotic-associated adverse events were rare. Primary Source of Funding: Agency for Healthcare Research and Quality (R01 HS026725).


Hematopoietic chimerism in sheep and nonhuman primates by in utero transplantation of fetal hematopoietic stem cells.
Bone marrow transplantation to reconstitute defective hematopoietic cell lines in children with congenital defects is limited by donor availability, graft rejection, and graft-versus-host disease (GVHD). These problems can be eliminated by transplanting normal preimmune fetal hematopoietic stem cells (HSC) into an unrelated preimmune fetal recipient. We show here that injections of allogeneic fetal stem cells into preimmune fetal lambs and monkeys result in long-term stable hematopoietic chimerism. HSCs harvested from the livers of preimmune fetal sheep and monkeys when injected into the peritoneal cavity of young unrelated fetal sheep and monkey recipients results in stable, long-term postnatal hematopoietic chimerism involving lymphoid, erythroid, and myeloid cells of donor origin. Donor cell engraftment was achieved without the use of cytoablative procedures and without the development of GVHD.


Nucleotide excision repair factor XPC enhances DNA damage-induced apoptosis by downregulating the antiapoptotic short isoform of caspase-2.
XPC protein is a critical DNA damage recognition factor in nucleotide excision repair for which genetic deficiency confers a predisposition to cancer. In this study, we show that XPC has a function that is independent of its canonical function in DNA repair, potentially altering the interpretation of how XPC deficiency leads to heightened cancer susceptibility. XPC enhances apoptosis induced by DNA damage in a p53 nullizygous background, acting downstream of mitochondrial permeabilization and upstream of caspase-9 activation in the DNA damage-induced apoptosis cascade. We found that deficiency in XPC upregulated production of the short isoform of caspase-2 (casp-2S). This upregulation occurred at both protein and mRNA levels through repression of the caspase-2 promoter by XPC protein. Targeted RNAi-mediated downregulation of casp-2S-enhanced UV-induced apoptosis as well as activation of caspase-9 and caspase-6 in XPC-deficient cells, but not in XPC-proficient cells. In addition, XPC overexpression in various p53-deficient cancer cells resistant to cisplatin improved their sensitivity to cisplatin-induced apoptosis. Given that casp-2S functions as an antiapoptotic protein, our findings suggest that XPC enhances DNA damage-induced apoptosis through inhibition of casp-2S transcription. Together, these findings offer a mechanistic foundation to overcome the resistance of highly prevalent p53-deficient tumors to cell death induced by DNA-damaging therapeutic agents, by targeting strategies that inhibit the expression or function of casp-2S.


[Percutaneous aortic aortic valve replacement: myth or reality?].
Aortic valve replacement with mechanical or biological prosthesis with extracorporeal circulation is the gold-standard for the treatment of calcific aortic stenosis. Although the results are excellent with the conventional approach some elderly patients, with multiple high-risk comorbid conditions, reoperations and severe left ventricular dysfunction have high surgical risk. During the last years percutaneous techniques have been developed. The present study aim to analyse the literature, since the experimental development until clinical application of this novel treatment in patients with high surgical risk aortic stenosis. Percutaneous implantation of aortic valve prosthesis is being done in some centers and the cardiovascular surgeon that treats valve disease should be involved in this development.


A preliminary proteomic characterisation of extracellular vesicles released by the ovine parasitic nematode, Teladorsagia circumcincta.
Teladorsagia circumcincta is a major cause of ovine parasitic gastroenteritis in temperate climatic regions. The development of high levels of anthelmintic resistance in this nematode species challenges its future control. Recent research indicates that many parasite species release extracellular vesicles into their environment, many of which have been classified as endocytic in origin, termed exosomes. These vesicles are considered to play important roles in the intercellular communication between parasites and their hosts, and thus represent potentially useful targets for novel control strategies. Here, we demonstrate that exosome-like extracellular vesicles can be isolated from excretory-secretory (ES) products released by T. circumcincta fourth stage larvae (Tci-L4ES). Furthermore, we perform a comparative proteomic analysis of vesicle-enriched and vesicle-free Tci-L4ES. Approximately 73% of the proteins identified in the vesicle-enriched fraction were unique to this fraction, whilst the remaining 27% were present in both vesicle-enriched and vesicle-free fraction. These unique proteins included structural proteins, nuclear proteins, metabolic proteins, proteolytic enzymes and activation-associated secreted proteins. Finally, we demonstrate that molecules present within the vesicles-enriched material are targets of the IgA and IgG response in T. circumcincta infected sheep, and could potentially represent useful targets for future vaccine intervention studies.


Corrigendum to: Does the C4 plant Trianthema portulacastrum (Aizoaceae) exhibit weakly expressed crassulacean acid metabolism (CAM)?
We examined whether crassulacean acid metabolism (CAM) is present in Trianthema portulacastrum L. (Aizoaceae), a pantropical, salt-tolerant C4 annual herb with atriplicoid-type Kranz anatomy in leaves but not in stems. The leaves of T. portulacastrum are slightly succulent and the stems are fleshy, similar to some species of Portulaca, the only genus known in which C4 and CAM co-occur. Low- level nocturnal acidification typical of weakly expressed, predominantly constitutive CAM was measured in plants grown for their entire life-cycle in an outdoor raised garden box. Acidification was greater in stems than in leaves. Plants showed net CO2 uptake only during the light irrespective of soil water availability. However, nocturnal traces of CO2 exchange exhibited curved kinetics of reduced CO2 loss during the middle of the night consistent with low-level CAM. Trianthema becomes the second genus of vascular land plants in which C4 and features of CAM have been demonstrated to co-occur in the same plant and the first C4 plant with CAM-type acidification described for the Aizoaceae. Traditionally the stems of herbs are not sampled in screening studies. Small herbs with mildly succulent leaves and fleshy stems might be a numerically significant component of CAM biodiversity.


Steroid hormone-related polymorphisms associate with the development of bone erosions in rheumatoid arthritis and help to predict disease progression: Results from the REPAIR consortium.
Here, we assessed whether 41 SNPs within steroid hormone genes associated with erosive disease. The most relevant finding was the rheumatoid factor (RF)-specific effect of the CYP1B1, CYP2C9, ESR2, FcγR3A, and SHBG SNPs to modulate the risk of bone erosions (P = 0.004, 0.0007, 0.0002, 0.013 and 0.015) that was confirmed through meta-analysis of our data with those from the DREAM registry (P = 0.000081, 0.0022, 0.00074, 0.0067 and 0.0087, respectively). Mechanistically, we also found a gender-specific correlation of the CYP2C9rs1799853T/T genotype with serum vitamin D3 levels (P = 0.00085) and a modest effect on IL1β levels after stimulation of PBMCs or blood with LPS and PHA (P = 0.0057 and P = 0.0058). An overall haplotype analysis also showed an association of 3 ESR1 haplotypes with a reduced risk of erosive arthritis (P = 0.009, P = 0.002, and P = 0.002). Furthermore, we observed that the ESR2, ESR1 and FcγR3A SNPs influenced the immune response after stimulation of PBMCs or macrophages with LPS or Pam3Cys (P = 0.002, 0.0008, 0.0011 and 1.97•10-7). Finally, we found that a model built with steroid hormone-related SNPs significantly improved the prediction of erosive disease in seropositive patients (PRF+ = 2.46•10-8) whereas no prediction was detected in seronegative patients (PRF- = 0.36). Although the predictive ability of the model was substantially lower in the replication population (PRF+ = 0.014), we could confirm that CYP1B1 and CYP2C9 SNPs help to predict erosive disease in seropositive patients. These results are the first to suggest a RF-specific association of steroid hormone-related polymorphisms with erosive disease.


[Peritonitis in diverticulitis: the Bern concept].
The colon is the most frequent origine for a diffuse peritonitis and diverticular perforation is again the most common source of a spontaneous secondary peritonitis. This paper first focuses on the treatment of peritonitis and secondly on the strategies of source control in peritonitis with special emphasis on the tactics (primary anastomosis vs. Hartmann procedure with colostomy) for surgical source control. Prospective analysis of 404 patients suffering from peritonitis (11/93-2/98), treated with an uniform treatment concept including early operation, source control and extensive intraoperative lavage (20 to 30 liters) as a standard procedure. Other treatment measures were added in special indications "on demand" only. Peritonitis was graded with the Mannheim Peritonitis Index (MPI). Tactics of source control in peritonitis due to diverticulitis were performed according to "general condition" respectively the MPI of the patient. The 404 patients averaged a MPI of 19 (0-35) in "local" peritonitis and a MPI of 26 (11-43) in "diffuse" peritonitis. The colon as a source of peritonitis resulted in MPI of 16 (0-33) in the case of "local" respectively 27 (11-43) in "diffuse" peritonitis. From 181 patients suffering from diverticulitis 144 needed an operation and in 78 (54%) peritonitis was present. Fourty-six percent (36) of the patients suffered from "local", 54% (42) from "diffuse" peritonitis. Resection with primary anastomosis was performed in 26% (20/78) whereas in 74% (58/78) of the patients a Hartmann procedure with colostomy was performed. The correlating MPI was 16 (0-28) vs. 23 (16-27) respectively. The analysis of complications and mortality based on the MPI showed a decent discrimination potential for primary anastomosis vs Hartmann procedure: morbidity 35% vs. 41%; reoperation 5% vs. 5%; mortality 0% vs. 14%. In case of peritonitis due to diverticulitis the treatment of peritonitis comes first. Thanks to advances in intensive care and improved anti-inflammatory care, a more conservative surgical concept nowadays is accepted. In the case of diverticulitis the MPI is helpful to choose between primary anastomosis vs. Hartmann procedure with colostomy as source control. The MPI includes the "general condition" of the patient into the tactical decision how to attain source control.


Prostaglandin E2-mediated attenuation of mesocortical dopaminergic pathway is critical for susceptibility to repeated social defeat stress in mice.
Various kinds of stress are thought to precipitate psychiatric disorders, such as major depression. Whereas studies in rodents have suggested a critical role of medial prefrontal cortex (mPFC) in stress susceptibility, the mechanism of how stress susceptibility is determined through mPFC remains unknown. Here we show a critical role of prostaglandin E(2) (PGE(2)), a bioactive lipid derived from arachidonic acid, in repeated social defeat stress in mice. Repeated social defeat increased the PGE(2) level in the subcortical region of the brain, and mice lacking either COX-1, a prostaglandin synthase, or EP1, a PGE receptor, were impaired in induction of social avoidance by repeated social defeat. Given the reported action of EP1 that augments GABAergic inputs to midbrain dopamine neurons, we analyzed dopaminergic response upon social defeat. Analyses of c-Fos expression of VTA dopamine neurons and dopamine turnover in mPFC showed that mesocortical dopaminergic pathway is activated upon social defeat and attenuated with repetition of social defeat in wild-type mice. EP1 deficiency abolished such repeated stress-induced attenuation of mesocortical dopaminergic pathway. Blockade of dopamine D1-like receptor during social defeat restored social avoidance in EP1-deficient mice, suggesting that disinhibited dopaminergic response during social defeat blocks induction of social avoidance. Furthermore, mPFC dopaminergic lesion by local injection of 6-hydroxydopamine, which mimicked the action of EP1 during repeated stress, facilitated induction of social avoidance upon social defeat. Taken together, our data suggest that PGE(2)-EP1 signaling is critical for susceptibility to repeated social defeat stress in mice through attenuation of mesocortical dopaminergic pathway.


Current status of three-dimensional echocardiography.
Volume rendered three-dimensional echocardiography (3DE), a rapidly emerging imaging method, offers clinicians and surgeons a new perspective for visualizing the heart. Using both transthoracic and transesophageal approach this new methodology enables qualitative and quantitative information of cardiac disorders to be obtained. 3DE image projections have allowed visualization of intracardiac structures and great vessels and delineation of their pathology in a comprehensive manner. Experimental and clinical studies suggest that 3DE is likely to play a valuable role in the evaluation of various cardiac and flow disorders and in planning interventions and surgery. In addition, 3DE has been applied to derive quantitative measurements of volume, mass and dimensions of the left and right ventricles and also other cardiac lesions, such as atrial and ventricular septal defects. Every aspect of 3DE is in continuous evolution. Faster and more highly automated image processing could make the technique more easily applicable in the clinical scenario.


Grafting synthetic transmembrane units to the engineered low-toxicity α-hemolysin to restore its hemolytic activity.
The chemical modification of proteins to provide desirable functions and/or structures broadens their possibilities for use in various applications. Usually, proteins can acquire new functions and characteristics, in addition to their original ones, via the introduction of synthetic functional moieties. Here, we adopted a more radical approach to protein modification, i.e., the replacement of a functional domain of proteins with alternative chemical compounds to build "cyborg proteins." As a proof of concept model, we chose staphylococcal α-hemolysin (Hla), which is a well-studied, pore-forming toxin. The hemolytic activity of Hla mutants was dramatically decreased by truncation of the stem domain, which forms a β-barrel pore in the membrane. However, the impaired hemolytic activity was significantly restored by attaching a pyrenyl-maleimide unit to the cysteine residue that was introduced in the remaining stem domain. In contrast, negatively charged fluorescein-maleimide completely abolished the remaining activity of the mutants.