Endocytic turnover of Rab8 controls cell polarization.
Adaptation of cell shape and polarization through the formation and retraction of cellular protrusions requires balancing of endocytosis and exocytosis combined with fine-tuning of the local activity of small GTPases like Rab8. Here, we show that endocytic turnover of the plasma membrane at protrusions is directly coupled to surface removal and inactivation of Rab8. Removal is induced by reduced membrane tension and mediated by the GTPase regulator associated with focal adhesion kinase-1 (GRAF1, also known as ARHGAP26), a regulator of clathrin-independent endocytosis. GRAF1-depleted cells were deficient in multi-directional spreading and displayed elevated levels of GTP-loaded Rab8, which was accumulated at the tips of static protrusions. Furthermore, GRAF1 depletion impaired lumen formation and spindle orientation in a 3D cell culture system, indicating that GRAF1 activity regulates polarity establishment. Our data suggest that GRAF1-mediated removal of Rab8 from the cell surface restricts its activity during protrusion formation, thereby facilitating dynamic adjustment of the polarity axis.


Sensorless freehand 3-D ultrasound using regression of the echo intensity.
In freehand 3-D ultrasound (US), a position sensor is attached to the probe of a 2-D US machine. The resulting 3-D data permit flexible visualisation and more accurate volume measurement than can be achieved using 2-D B-scans alone; however, the use of the position sensor can be inconvenient for the clinician. The objective is, thus, to replace the sensor with a technique for estimating the probe trajectory based on the B-scan images, themselves. One such technique exists, based on decorrelation algorithms. This paper presents an alternative approach based on linear regression of the echo-envelope intensity signal. A probabilistic analysis of the speckle characteristics of the US signal leads to a linear model, on which the regression algorithm is based. The gradient parameter of this model is shown to be directly related to probe motion. The viability of the new approach is demonstrated through simulations and in vitro and in vivo experiments.


Knowledge is power: Relationship between professional recommendations, maternal attitudes, and screen time exposure of infants.
The American Academy of Pediatrics (AAP) and the World Health Organization (WHO) recommend infants avoid screen media exposure, yet most infants are regularly exposed. This study aimed to explore screen exposure, maternal attitudes regarding screen media effects, and pediatricians' recommendations to better understand widespread screen media use with infants younger than 18 months of age in hopes of informing mitigation efforts. Surveys consisting of 10 Likert-style questions were distributed real-time to a purposive sample of 193 mothers with infants 18 months of age or younger in Georgia, Pennsylvania, and New York. Seventy-nine percent of infants exceeded AAP/WHO guidelines and 61% of respondents couldn't recall receiving pediatrician recommendations regarding screen exposure for their infant. Mothers with higher levels of education were associated with an accurate reflection of the adverse effects of screen usage on infant development (ANOVA; F = 10.122; df = 3; p < .001). Accurate maternal attitudes regarding adverse effects on infants was associated with less daily screen exposure (Spearman correlation; r = -0.428; p < .001). Findings suggest that knowledge about adverse side effects of screen media usage with infants is associated with less daily exposure; and, that pediatricians may not routinely review guidelines for infants with their families, underscoring the importance of other methods of knowledge dissemination. Nurses, as trusted healthcare providers, can provide counseling on infant screen media usage creating an opportunity for augmented knowledge dissemination across maternal demographics potentially leading to less screen exposure in this vulnerable population.


Phosphonate as a Stable Zinc-Binding Group for "Pathoblocker" Inhibitors of Clostridial Collagenase H (ColH).
Microbial infections are a significant threat to public health, and resistance is on the rise, so new antibiotics with novel modes of action are urgently needed. The extracellular zinc metalloprotease collagenase H (ColH) from Clostridium histolyticum is a virulence factor that catalyses tissue damage, leading to improved host invasion and colonisation. Besides the major role of ColH in pathogenicity, its extracellular localisation makes it a highly attractive target for the development of new antivirulence agents. Previously, we had found that a highly selective and potent thiol prodrug (with a hydrolytically cleavable thiocarbamate unit) provided efficient ColH inhibition. We now report the synthesis and biological evaluation of a range of zinc-binding group (ZBG) variants of this thiol-derived inhibitor, with the mercapto unit being replaced by other zinc ligands. Among these, an analogue with a phosphonate motif as ZBG showed promising activity against ColH, an improved selectivity profile, and significantly higher stability than the thiol reference compound, thus making it an attractive candidate for future drug development.


Acute particulate matter affects cardiovascular autonomic modulation and IFN-γ methylation in healthy volunteers.
Air particulate matter (PM) is associated with increased cardiovascular morbidity and mortality. Altered autonomic functions play a key role in PM-induced cardiovascular disease. However, previous studies have not address the impact of PM on sympathetic and parasympathetic control of heart function, independently, and using controlled conditions, i.e., increasing titration of PM of known composition, in absence of other potential confounding factors. To fill this gap, here we used symbolic analysis that is capable of detecting non-mutual changes of the two autonomic branches, thus considering them as independent, and concentrations of PM as they could be measured at peak levels in Milan during a polluted winter day. In this randomized, cross-over study, we enrolled 12 healthy subjects who underwent two random sessions: inhalation of filtered air mixture or inhalation of filtered air containing particulate mixture (PM 10, PM 2.5, PM 1.0 and PM 0.5µm). ECG and respiration for autonomic analysis and blood sample for DNA Methylation were collected at baseline (T1), after air exposure (T2) and after 2h (T3). Spectral and symbolic analysis of heart rate variability (HRV) were performed for autonomic control of cardiac function, while alterations in DNA methylation of candidate genes were used to index pro-inflammatory modifications. In the PM expose group, autonomic analysis revealed a significant decrease of 2UV%, index of parasympathetic modulation (14% vs 9%, p = 0.0309), while DNA analysis showed a significant increase of interferon γ (IFN- γ) methylation, from T1 to T3. In a mixed model using T1, T2 and T3, fine and ultrafine PM fractions showed significant associations with IFN- γ methylation and parasympathetic modulation. Our study shows, for the first time, that in healthy subjects, acute exposure to PM affects parasympathetic control of heart function and it increases methylation of a pro-inflammatory gene (i.e. methylation of interferon γ). Thus, our study suggests that, even in absence of other co-factors and in otherwise healthy individuals, PM per se is sufficient to trigger parasympathetic dysautonomia, independently from changes in sympathetic control, and inflammation, in a dose-dependent manner.


Tolevamer, a novel nonantibiotic polymer, compared with vancomycin in the treatment of mild to moderately severe Clostridium difficile-associated diarrhea.
Current antibiotic therapies for Clostridium difficile-associated diarrhea have limitations, including progression to severe disease, recurrent C. difficile-associated diarrhea, and selection for nosocomial pathogens. Tolevamer, a soluble, high-molecular weight, anionic polymer that binds C. difficile toxins A and B is a unique nonantibiotic treatment option. In this 3-arm, multicenter, randomized, double-blind, active-controlled, parallel-design phase II study, patients with mild to moderately severe C. difficile-associated diarrhea were randomized to receive 3 g of tolevamer per day (n = 97), 6 g of tolevamer per day (n = 95), or 500 mg of vancomycin per day (n = 97). The primary efficacy parameter was time to resolution of diarrhea, defined as the first day of 2 consecutive days when the patient had hard or formed stools (any number) or < or = 2 stools of loose or watery consistency. In the per-protocol study population, resolution of diarrhea was achieved in 48 (67%) of 72 patients receiving 3 g of tolevamer per day (median time to resolution of diarrhea, 4.0 days; 95% confidence interval, 2.0-6.0 days), in 58 (83%) of 70 patients receiving 6 g of tolevamer per day (median time to resolution of diarrhea, 2.5 days; 95% confidence interval, 2.0-3.0 days), and in 73 (91%) of 80 patients receiving vancomycin (median time to resolution of diarrhea, 2.0 days; 95% confidence interval, 1.0-3.0 days). Tolevamer administered at a dosage of 6 g per day was found to be noninferior to vancomycin administered at a dosage of 500 mg per day with regard to time to resolution of diarrhea (P = .02) and was associated with a trend toward a lower recurrence rate. Tolevamer was well tolerated but was associated with an increased risk of hypokalemia. Tolevamer, a novel polystyrene binder of C. difficile toxins A and B, effectively treats mild to moderate C. difficile diarrhea and merits further clinical development.


Wearable Fiber Optic Technology Based on Smart Textile: A Review.
Emerging smart textiles have enriched a variety of wearable technologies, including fiber optic technology. Optic fibers are widely applied in communication, sensing, and healthcare, and smart textiles enable fiber optic technology to be worn close to soft and curved human body parts for personalized functions. This review briefly introduces wearable fiber optic applications with various functions, including fashion and esthetics, vital signal monitoring, and disease treatment. The main working principles of side emission, wavelength modulation, and intensity modulation are summarized. In addition, textile fabrication techniques, including weaving and knitting, are discussed and illustrated as combination methods of embedding fiber optic technology into textile fabric. In conclusion, the combination of optical fibers and textiles has drawn considerable interest and developed rapidly. This work provides an overview of textile-based wearable fiber optic technology and discusses potential textile fabrication techniques for further improvement of wearable fiber optic applications.


Influence of the novel histamine H₃ receptor antagonist ST1283 on voluntary alcohol consumption and ethanol-induced place preference in mice.
Growing evidence supports a role for the central histaminergic system to have a modulatory influence on drug addiction in general and alcohol-use disorders in particular through histamine H3 receptors (H3R). In the present study, the effects of systemic injection of the newly synthesized H3R antagonist ST1283 on ethanol (EtOH) voluntary intake and EtOH-conditioned reward in mice have been investigated. Oral EtOH, saccharin, and quinine intake was assessed in a two-bottle choice paradigm using escalating concentrations of alcohol or tastant solutions. EtOH-induced place preference (CPP), EtOH-induced locomotor activity, and blood ethanol concentration (BEC) were also measured. Following administration of the H3R antagonist (2.5, 5, and 10 mg/kg, i.p.), there was a significant dose-dependent decrease in alcohol consumption and preference. Importantly, vehicle- and ST1283 (5 mg/kg)-treated mice showed similar consumption and preference to increasing concentration of both sweet and bitter tastes. More interestingly, systemic administration of ST1283 inhibited EtOH-CPP and EtOH-enhanced locomotion. This inhibition was blocked when mice were pretreated with the selective H3R agonist R-(alpha)-methyl-histamine (10 mg/kg). Finally, vehicle- and ST1283-treated mice had similar BECs. Our results show that ST1283 may decrease voluntary EtOH consumption and EtOH-CPP by altering its reinforcing effects, suggesting a novel role for histamine signaling in regulation of alcoholism. Lastly, the results add to the growing literature on H3R modulation in the pharmacotherapy of EtOH addiction.


Efficacy of treatment for hyperglycemic crisis in elderly diabetic patients in a day hospital.
The purpose of this prospective cohort study was to compare the costs of day hospital (DH) care for hyperglycemic crisis in elderly diabetic patients with those of conventional hospitalization (CH). Secondary objectives were to compare these two clinical scenarios in terms of glycemic control, number of emergency and outpatient visits, readmissions, hypoglycemic episodes, and nosocomial morbidity. The study population comprised diabetic patients aged >74 years consecutively admitted to a tertiary teaching hospital in Spain for hyperglycemic crisis (sustained hyperglycemia [>300 mg/dL] for at least 3 days with or without ketosis). The patients were assigned to DH or CH care according to time of admission and were followed for 6 months after discharge. Exclusion criteria were ketoacidosis, hyperosmolar crisis, hemodynamic instability, severe intercurrent illness, social deprivation, or Katz index >D. Sixty-four diabetic patients on DH care and 36 on CH care were included, with no differences in baseline characteristics. The average cost per patient was 1,345.1±793.6 € in the DH group and 2,212.4±982.5 € in the CH group (P<0.001). There were no differences in number of subjects with mild hypoglycemia during follow-up (45.3% DH versus 33.3% CH, P=0.24), nor in the percentage of patients achieving a glycated hemoglobin (HbA(1c)) <8% (67.2% DH versus 58.3% CH, P=0.375). Readmissions for hyperglycemic crisis and pressure ulcer rates were significantly higher in the CH group. DH care for hyperglycemic crises is more cost-effective than CH care, with a net saving of 1,418.4 € per case, lower number of readmissions and pressure ulcer rates, and similar short-term glycemic control and hypoglycemia rates.


Relationships of active smoking to asthma and asthma severity in the EGEA study. Epidemiological study on the Genetics and Environment of Asthma.
The role of smoking as potential risk factor, selection factor ("healthy smoker" effect) and modifying factor (severity) of asthma was studied in the Epidemiological study on the Genetics and Environment of Asthma, bronchial hyperresponsiveness and atopy (EGEA). The analysis involved 200 adult asthmatic cases recruited in chest clinics, 265 nonasthmatic controls and 586 relatives of asthmatics (147 with asthma). Asthma in childhood was not associated with a reduced take-up of smoking (odds ratio (OR)=1.06 in males and 0.98 in females), but smoker asthmatic cases quit more often than controls (OR = 2.20 (95% confidence interval (95% CI) 1.11-4.34) in males and 2.76 (1.19-6.42) in females). Adult onset asthma was unrelated to ever smoking (OR 1.07 in males and 1.02 in females). In asthmatic cases, active smoking was associated with asthma severity. Current smokers, compared to never and exsmokers, had more asthma symptoms, more frequent (> or =1 attack x day(-1)) asthma attacks (OR 2.39 (95% CI 1.06-5.36)) and higher asthma severity scores. No clear pattern regarding the relationships of smoking habits with asthma was observed in first degree relatives. It is concluded that active smoking is not a risk factor for asthma in adulthood, but that smoking increases asthma severity.


Biophysical, Biochemical, and Cell Based Approaches Used to Decipher the Role of Carbonic Anhydrases in Cancer and to Evaluate the Potency of Targeted Inhibitors.
Carbonic anhydrases (CAs) are thought to be important for regulating pH in the tumor microenvironment. A few of the CA isoforms are upregulated in cancer cells, with only limited expression in normal cells. For these reasons, there is interest in developing inhibitors that target these tumor-associated CA isoforms, with increased efficacy but limited nonspecific cytotoxicity. Here we present some of the biophysical, biochemical, and cell based techniques and approaches that can be used to evaluate the potency of CA targeted inhibitors and decipher the role of CAs in tumorigenesis, cancer progression, and metastatic processes. These techniques include esterase activity assays, stop flow kinetics, and mass inlet mass spectroscopy (MIMS), all of which measure enzymatic activity of purified protein, in the presence or absence of inhibitors. Also discussed is the application of X-ray crystallography and Cryo-EM as well as other structure-based techniques and thermal shift assays to the studies of CA structure and function. Further, large-scale genomic and proteomic analytical methods, as well as cell based techniques like those that measure cell growth, apoptosis, clonogenicity, and cell migration and invasion, are discussed. We conclude by reviewing approaches that test the metastatic potential of CAs and how the aforementioned techniques have contributed to the field of CA cancer research.


Calcium intake, calcium bioavailability and bone health.
Calcium accounts for 1-2 % of adult human body weight. Over 99 % of total body Ca is found in the teeth and bones. Therefore, in addition to the obvious structural role of the skeleton, it also serves as a reservoir for Ca. Dietary Ca intake has an important impact on bone metabolism and bone health. Chronic Ca deficiency resulting from inadequate intake or poor intestinal absorption is one of several important causes of reduced bone mass and osteoporosis. It is vital, therefore, that adequate dietary Ca is consumed at all stages of life - in early life so that the genetically programmed peak bone mass can be reached and in later adulthood so that the skeletal mass can be maintained and age-related bone loss minimised. Unfortunately, there is wide variation in the estimates of daily Ca requirements made by different expert authorities. Furthermore, there is evidence that many individuals are not consuming these recommended levels. The consequence of this for bone health will be discussed in the present review. Besides the amount of Ca in the diet, the absorption of dietary Ca in foods is also a critical factor in determining the availability of Ca for bone development and maintenance. Thus, there is a need to identify food components and/or functional food ingredients that may positively influence Ca absorption in order to ensure that Ca bioavailability from foods can be optimised. This approach may be of particular value in individuals who fail to achieve the dietary recommended level of Ca.


[The role of delta-sleep-inducing peptide in the normalization of cardiac electric stability in experimental emotional stress].
In chronic experiments on rabbits, a delta-sleeping inducing peptide was tested for effects on the cardiac electric stability and onset of ventricular premature beats during experimental emotional stress. The peptide in a dose of 60 nm/kg was found to normalize the thresholds of ventricular fibrillation and its predictors and to decrease or abolish spontaneous ventricular premature beats occurring during the stress. This suggests that the delta-sleep-inducing peptide may be used to prevent cardiac arrhythmias during stresses.


[Causal treatment of corneal vascularization (author's transl)].
Corneal vascularization is attributable to the swelling and increased lactate concentration in the cornea, which are caused by damage to the oxidizing metabolism. Invasion of leukocytes stimulated by prostaglandins also promotes the increase in the lactate concentration. The following causal treatment is possible: 40% glucose to combat the swelling, 3% cystein to improve the oxidizing metabolism and counteract the increased lactate concentration, and 0.5% indomethacin suspension to prevent the formation of prostaglandins and hence the invasion of leukozytes.


Endotoxin-induced renal failure. II. A role for tubular hypoxic damage.
Endotoxin-induced hypotension and altered renal microcirculation could lead to tubular injury, particularly at the physiologically hypoxic outer medulla. We explored this hypothesis in isolated perfused kidneys and in vivo in rats subjected to endotoxemia. Rat kidneys were removed 15 min after endotoxin injection in vivo (from Escherichia coli 0127:B8, 1 mg/kg i.p.) and perfused with oxygenated medium supplemented with 20 amino acids and endotoxin. Glomerular filtration rate and filtration fraction markedly declined (0.4 +/- 0. 1 ml/min and 1.1 +/- 0.1, respectively) as compared with control kidneys (0.7 +/- 0.1 ml/min and 1.8 +/- 0.1, n = 8-12 per group; p < 0.05). Hypoxic injury to medullary thick ascending limbs in the innermost outer medulla increased (47 +/- 9% of tubules vs. 16 +/- 8% in controls, p < 0.05). When rats were preconditioned with an additional endotoxin injection 16 h earlier (a manipulation that markedly reduces cortical and medullary blood flow), glomerular filtration rate and filtration fraction further declined to 0.1 +/- 0.0 ml/min and 0.4 +/- 0.1, respectively (p < 0.01), and tubular sodium reabsorption fell to 81 +/- 12 vs 98 +/- 0% in controls (p < 0.05). Tubular damage, however, did not increase (20 +/- 7%), probably reflecting a decline in reabsorptive workload and oxygen requirement. In rats subjected to a single or two repeated daily doses of endotoxin (1 mg/kg i.p.) plasma creatinine comparably rose 41% on the average over 24 h, creatinine clearance fell by 27% (p < 0.0001), but tubular damage was absent. By contrast, in rats preconditioned with indomethacin and the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (10 mg/kg), the addition of endotoxin markedly augmented outer medullary hypoxic tubular damage both in S(3) segments (27 +/- 10 vs 1 +/- 1%) and in medullary thick ascending limbs (38 +/- 11 vs. 10 +/- 5%, n = 7-8; p < 0.05). It is concluded that under special conditions, such as altered medullary oxygen balance or defective nitric oxide or prostaglandin synthesis, endotoxin may predispose to hypoxic outer medullary tubular damage.


Poor reliability of vaginal and endocervical cultures for evaluating microbiology of endometrial cavity in women with chronic endometritis.
Chronic endometritis (CE) is a subtle pathology causing infertility and abnormal uterine bleeding. We evaluated the reliability of vaginal and cervical cultures for detecting infectious agents at the endometrial level. In a prospective diagnostic study, 181 women diagnosed with CE and 100 controls underwent vaginal, endocervical and endometrial sampling. Cultures for common bacteria, Neisseria gonorrhoeae, yeast and Ureaplasma urealyticum and PCR for Chlamydia trachomatis were performed. The prevalent infectious agents at the endometrial level were common bacteria(59.7% of cases); U. urealyticum was detected in 11.0% and C. trachomatis in only 2.8%. The concordance rate between endocervical and endometrial specimens for common bacteria was 48.3%; 100% for C. trachomatis and 58.3% for U. urealyticum. The concordance rate between vaginal and endometrial cultures for common bacteria was 50.2%, only 16.7% for C. trachomatis and 48.8% for U. urealyticum. For common bacteria both vaginal and cervical cultures showed low sensitivities of 0.30 and 0.19, respectively. Common bacteria and U. urealyticum were the prevalent infectious agents in the uterine cavity of women diagnosed with CE. Both vaginal and endocervical cultures had low concordance with endometrial cultures. Only C. trachomatis test at cervical level had high concordance with endometrial findings.


DYRK1A suppression restrains Mcl-1 expression and sensitizes NSCLC cells to Bcl-2 inhibitors.
Objective: Mcl-1 overexpression confers acquired resistance to Bcl-2 inhibitors in non-small cell lung cancer (NSCLC), but no direct Mcl-1 inhibitor is currently available for clinical use. Thus, novel therapeutic strategies are urgently needed to target Mcl-1 and sensitize the anti-NSCLC activity of Bcl-2 inhibitors. Methods: Cell proliferation was measured using sulforhodamine B and colony formation assays, and apoptosis was detected with Annexin V-FITC staining. Gene expression was manipulated using siRNAs and plasmids. Real-time PCR and Western blot were used to measure mRNA and protein levels. Immunoprecipitation and immunofluorescence were used to analyze co-localization of dual specificity tyrosine-phosphorylation-regulated kinase 1A (DYRK1A) and Mcl-1. Results: Suppression of DYRK1A resulted in reduced Mcl-1 expression in NSCLC cells, whereas overexpression of DYRK1A significantly increased Mcl-1 expression. Suppression of DYRK1A did not alter Mcl-1 mRNA levels, but did result in an accelerated degradation of Mcl-1 protein in NSCLC cells. Furthermore, DYRK1A mediated proteasome-dependent degradation of Mcl-1 in NSCLC cells, and DYRK1A co-localized with Mcl-1 in NSCLC cells and was co-expressed with Mcl-1 in tumor samples from lung cancer patients, suggesting that Mcl-1 may be a novel DYRK1A substrate. We showed that combined therapy with harmine and Bcl-2 antagonists significantly inhibited cell proliferation and induced apoptosis in NSCLC cell lines as well as primary NSCLC cells. Conclusions: Mcl-1 is a novel DYRK1A substrate, and the role of DYRK1A in promoting Mcl-1 stability makes it an attractive target for decreasing Bcl-2 inhibitor resistance.


An experimental and theoretical investigation of loperamide hydrochloride-glutaric acid cocrystals.
Cocrystallization is a powerful method to improve the physicochemical properties of drugs. Loperamide hydrochloride is a topical analgesic for the gastrointestinal tract showing low and pH-dependent solubility; for this reason, an enhancement of its solubility or dissolution rate, particularly at the pH of the intestinal tract, could improve its local efficacy. Here we prepared cocrystals of this active principle with glutaric acid and so obtained a new crystalline solid representing a viable alternative to improve the physicochemical properties and thus the pharmaceutical behavior of the drug. Differential scanning calorimetry, X-ray powder diffraction, Fourier infrared spectroscopy, solid-state NMR, and scanning electron microscopy coupled to the energy-dispersive X-ray spectrometry were used to investigate the new solid-phase formation. DFT calculations at B3LYP/6-31G(d) level of theory, in the gas phase, including frequencies computation, provided a rationale for the interaction between loperamide hydrochloride and glutaric acid. The cocrystals showed improved water solubility in comparison with loperamide HCl, and the pharmaceutical formulation proposed was able to release the drug more rapidly in comparison with three reference commercial products when tested at neutral pH values.


Microcystin ecotypes in a perennial Planktothrix agardhii bloom.
The dynamics and microcystins (MC) concentrations of a perennial Planktothrix agardhii bloom were investigated in a eutrophic lake (Viry-Châtillon, France). A weak relationship was observed between P. agardhii population biomass and the MC concentrations in a 1-year survey. To further investigate the causes of MC concentration changes, we concurrently conducted experiments on 41 strains isolated from this lake. We first checked the clonal diversity of P. agardhii population (i) by molecular techniques, to assess the presence of MC synthetase gene (mcyB), (ii) by biochemical assay (PP2A inhibition assay), for MC production, and (iii) by mass spectrometry (MS), to identify the MC chemotypes. Our results illustrated the diversity of genotype and MC chemotypes within a P. agardhii natural population. Eleven chemotypes among the 16 possible ones were found by MS. Furthermore, we noticed major differences in the MC content of isolated strains (from 0.02 to 1.86 microg equiv. MC-LR mg DW(-1), n=25). Growth and MC production of one MC-producing strain and one non-MC-producing strain were also assessed at two temperatures (10 and 20 degrees C). We showed that growth capacities of these strains were similar at the two tested temperatures, and that the MC production rate was correlated to the growth rate for the MC-producing strain. On the basis of these results, several hypotheses are discussed to explain the weakness of relationships between natural P. agardhii biomass and MC concentration. One of the main reasons could lie in the proportion of MC-producing clones and non-MC-producing clones that may change during the sampling period. Also, the MC-producing clones may present different intracellular MC content due to (i) MC chemotypes diversity, (ii) changes in MC variants proportions within a strain, and (iii) changes in MC rate production depending on the physiological state of cells. Finally, we concluded that various biological organization levels have to be considered (population, cellular and molecular), through an integrative approach, in order to provide a better understanding of P. agardhii in situ MC production.


Adaptive filter design via a gradient thresholding algorithm for compressive spectral imaging.
Sensing a spectral image data cube has traditionally been a time-consuming task since it requires a scanning process. In contrast, compressive spectral imaging (CSI) has attracted widespread interest since it requires fewer samples than scanning systems to acquire the data cube, thus improving the sensing speed. CSI captures linear projections of the scene, and then a reconstruction algorithm estimates the underlying scene. One notable CSI architecture is the color coded aperture snapshot spectral imager (C-CASSI), which employs pixelated filter arrays as the coding patterns to spatially and spectrally encode the incoming light. Up to date works on C-CASSI have used non-adaptive color coded apertures. Non-adaptive sampling ignores prior information about the signal to design the coding patterns. Therefore, this work proposes a method to adaptively design the color coded aperture, such that the quality of image reconstruction is improved. In more detail, this work introduces a gradient thresholding algorithm, which computes the consecutive color coded aperture from a rapidly reconstructed low-resolution version of the data cube. The successive adaptive patterns enable recovering a data cube in the presence of Gaussian noise with higher image quality. Real reconstructions and simulations evidence an improvement of up to 3 dB in the quality of image reconstruction of the proposed method in comparison with state-of-the-art non-adaptive techniques.


Vibrio cholerae OmpR Contributes to Virulence Repression and Fitness at Alkaline pH.
Vibrio cholerae is a Gram-negative human pathogen and the causative agent of the life-threatening disease cholera. V. cholerae is a natural inhabitant of marine environments and enters humans through the consumption of contaminated food or water. The ability to transition between aquatic ecosystems and the human host is paramount to the pathogenic success of V. cholerae The transition between these two disparate environments requires the expression of adaptive responses, and such responses are most often regulated by two-component regulatory systems such as the EnvZ/OmpR system, which responds to osmolarity and acidic pH in many Gram-negative bacteria. Previous work in our laboratory indicated that V. cholerae OmpR functioned as a virulence regulator through repression of the LysR-family transcriptional regulator aphB; however, the role of OmpR in V. cholerae biology outside virulence regulation remained unknown. In this work, we sought to further investigate the function of OmpR in V. cholerae biology by defining the OmpR regulon through RNA sequencing. This led to the discovery that V. choleraeompR was induced at alkaline pH to repress genes involved in acid tolerance and virulence factor production. In addition, OmpR was required for V. cholerae fitness during growth under alkaline conditions. These findings indicate that V. cholerae OmpR has evolved the ability to respond to novel signals during pathogenesis, which may play a role in the regulation of adaptive responses to aid in the transition between the human gastrointestinal tract and the marine ecosystem.


Effect of polymorph derived oxidants on IgG in relation to rheumatoid factor binding.
Immunoglobulin G from rheumatoid patients is denatured around the hinge region. This has been proposed as an explanation for the presence of circulating autoantibodies to IgG in these patients. It has previously been suggested that oxygen radicals (OR) derived from activated polymorphs may play a role in denaturation in vivo. Using sera from rheumatoid patients and age-matched controls in a modified ELISA technique, we have investigated the potential for polyclonal rheumatoid factors (RF) to bind to OR denatured IgG. Three model systems were used to generate OR in vitro: (a) purified PMN s activated by the cell surface stimulant PMA, (b) radiolysis of IgG in solution to generate specifically the superoxide radical and, in a separate system, the hydroxyl radical, (OH.), (c) purified myeloperoxide in the presence of H2O2 and halide ions. 1. The binding of both IgA and IgM RF s to PMN denatured IgG increased dose dependently for seropositive sera only. 2. The OH. radical but not the superoxide radical significantly increased the binding of IgA and M RF, again only for seropositive sera. 3. The myeloperoxidase enzyme system did not increase RF binding. 4. IgG incubated with elastase was not found to be a better antigen than native IgG. These results indicate that IgG is denatured by OR released from activated PMN, thereby producing an antigen for polyclonal RF s.


Phenotypic determinism and stochasticity in antibody repertoires of clonally expanded plasma cells.
The capacity of humoral B cell-mediated immunity to effectively respond to and protect against pathogenic infections is largely driven by the presence of a diverse repertoire of polyclonal antibodies in the serum, which are produced by plasma cells (PCs). Recent studies have started to reveal the balance between deterministic mechanisms and stochasticity of antibody repertoires on a genotypic level (i.e., clonal diversity, somatic hypermutation, and germline gene usage). However, it remains unclear if clonal selection and expansion of PCs follow any deterministic rules or are stochastic with regards to phenotypic antibody properties (i.e., antigen-binding, affinity, and epitope specificity). Here, we report on the in-depth genotypic and phenotypic characterization of clonally expanded PC antibody repertoires following protein immunization. We find that clonal expansion drives antigen specificity of the most expanded clones (top ∼10), whereas among the rest of the clonal repertoire antigen specificity is stochastic. Furthermore, we report both on a polyclonal repertoire and clonal lineage level that antibody-antigen binding affinity does not correlate with clonal expansion or somatic hypermutation. Last, we provide evidence for convergence toward targeting dominant epitopes despite clonal sequence diversity among the most expanded clones. Our results highlight the extent to which clonal expansion can be ascribed to antigen binding, affinity, and epitope specificity, and they have implications for the assessment of effective vaccines.


Rap1-B is phosphorylated by protein kinase A in intact human platelets.
Agonists that increase cAMP levels in platelets promote the phosphorylation of a 24 kDa GTP-binding protein that is immunoreactive with a monoclonal antibody (M90) to the H-ras p21 protein. Evidence is presented which indicates that this protein is rap-1b, not rap1-a as previously suggested (Ohmori, T., Kikuchi, A., Yamamoto, K., Kawata, M., Kondo, J. and Takai, Y. (1988) Biochem. Biophys. Res. Commun. 157, 670-676). The amino acid sequence of labeled peptides obtained by proteolytic cleavage of the purified phosphorylated protein was identical with that of rap-1b. Furthermore, a comparison of the kinetics of phosphorylation of synthetic peptides corresponding to the C-terminal region of rap-1a and rap-1b proteins indicated that rap-1b is the preferred substrate for phosphorylation by cAMP-dependent protein kinase.


Hepatitis B virus infection testing and prevalence among Asian and Pacific Islanders.
Asian and Pacific Islanders (APIs) constitute less than 6% of the US population, but account for more than half of Americans with chronic hepatitis B virus (HBV) infection. We sought to examine the effect of country of origin on HBV testing and chronic HBV infection prevalence among APIs. We analyzed demographic and clinical data collected for adults from Kaiser Permanente Hawaii with 1 or more healthcare encounters during 2006 to 2008, 12 months or more of follow-up before 2009, and no HBV-related diagnosis within 6 months of enrollment. Persons who received a test and a positive test result for HBV surface antigen or HBV DNA were classified "tested" and with "chronic HBV infection," respectively. Of 92,687 eligible APIs, 53,573 (58%) had country-of-origin data available. Among those, 41,263 were US born; 28.3% were tested; and 1.8% of those tested had chronic HBV infection. Of 12,310 foreign-born APIs, 30.5% were tested and 7.4% of those tested had chronic HBV infection. Foreignborn APIs had higher odds of being tested (odds ratio [OR] = 1.15) and testing positive (OR = 4.18) compared with US-born APIs. Persons with 2 or more abnormal tests for alanine aminotransferase (ALT) levels had higher odds of getting tested (OR = 6.12) and of testing positive (OR = 1.86) compared with persons with other ALT levels. Less than one-third of this managed care API population (29% of 53,573) was tested, yet the prevalence of chronic HBV infection (3.2%) was 12 times higher than that of the general US population. These findings underscore the importance of adherence to HBV testing guidelines to identify persons with infection so they may be linked to care.


Effect of Diameter of Saphenous Vein on Stump Length after Radiofrequency Ablation for Varicose Vein.
Radiofrequency ablation (RFA) has gained popularity for treatment of varicose veins. The diameter of the saphenous vein should be considered before RFA because occlusion of the vein may differ depending on its diameter. Until now, however, there have been few data about the correlation between the diameter of the saphenous vein and the stump length after RFA. The purpose of our study was to investigate its correlation. A retrospective review was performed from prospectively collected data of RFA patients between March 2009 and December 2011. Preoperatively, the saphenous vein diameter was measured. Ablation was initiated 2 cm distal from the junction. Postoperatively, stump length was measured at 1 week and 6 months. After 2 years, we measured the length from the saphenofemoral junction to the leading point of occlusion for great saphenous vein, and length from the saphenopopliteal junction to the leading point of occlusion for small saphenous vein. The paired t-test, independent t-test, and correlation analysis were used for statistical analysis. P-value <0.05 was considered statistically significant. During the study period, RFA was performed in 201 patients. Endovenous heat-induced thrombosis developed in 3 patients (1.5%). After 2 years, the stump length was obtained in 74 limbs. The mean diameter and stump length of the saphenous vein were 6.7±1.8 mm and 12.5±8.5 mm, respectively. Correlation analysis showed that the Pearson correlation coefficient of these factors was -0.017. There was no correlation between the diameter of saphenous vein and stump length.


Copulation duration and fertilization success in a damselfly: an example of cryptic female choice?
Copulation duration is highly variable (0.5-3 h) in the damselfly, Ceriagrion tenellum (Coenagrionidae). Using laboratory experiments, we tested four adaptive hypotheses to explain this variation: the effect of time constraints, in-copula mate guarding, sperm displacement and cryptic female choice. Copulation duration was negatively correlated with time of day, as predicted by the first two hypotheses, and positively correlated with male density, as predicted by the mate-guarding hypothesis. Males prolonged copulation in response to the volume of sperm stored by females, suggesting they were able to detect and quantify the amount of sperm stored. This behaviour is not explained by mate guarding or time constraint effects. Males removed all the sperm from the bursa copulatrix in just 10 min. Our results also suggest that, because the duct is too narrow to allow male genitalia to enter, males do not remove spermathecal sperm. Therefore, direct sperm removal could not explain long copulations. Prolonged copulations could also have evolved as a result of cryptic female choice if they increase male fertilization success by female-mediated processes. Our results support this idea: male fertilization success was greater after long copulations. Apparently, male copulatory behaviour elicits female responses that increase male fertilization success. Copyright 2000 The Association for the Study of Animal Behaviour.


Primary intramuscular cold abscess in the left deltoid region: a case report.
A case of primary intramuscular cold abscess in the left deltoid region is described. There was no underlying bone lesion in relation to cervical or thoracic spine, shoulder joint or upper humerus, and clavicle. It is postulated that the cold abscess might have developed as a result of reactivation of BCG lesion when the patient received a fresh dose of tubercular bacilli which produced only lymph node enlargement in the mediastinum because of the partial immunity the patient had.


Introital ultrasound in the diagnosis of lower urinary tract symptoms following anti-incontinence surgery using a synthetic midurethral tape.
Surgical treatment of stress urinary incontinence in women using a synthetic midurethral tape has become a standard procedure. One of the complications observed postoperatively are lower urinary tract symptoms (LUTS). The aim was to analyze the role of introital ultrasound in the identification of patients at risk for developing LUTS after surgical treatment using synthetic tape. A group of 50 patients suffering from LUTS following anti-incontinence surgery using synthetic tape was included in this study. The patients with pelvic organ prolapse and coexisting overactive bladder-wet before surgery were excluded. The control group consisted of 50 patients after the same treatment without any complications and with a good outcome. Tape visualization was performed using introital two-dimensional ultrasound. The assessment of the Tape Index (T/U) enabled us to divide the study group into the two subgroups with the tape index value of 0.375 as a borderline. The correlation between the tape position and the occurrence of LUTS was evaluated using a Chi-squared test. In the group of patients suffering from LUTS, the tape was found to be closer to the bladder neck (the lower edge of the tape was more than 37.5% of the urethral length) and it was statistically significant (Chi-squared = 19.87, p < 0.001). The tape position in the proximal urethra may have an impact on the postoperative occurrence of LUTS. The simple method of introital ultrasound could allow the identification of patients at risk for the development of LUTS after anti-incontinence surgery using synthetic tape.


Phenotypic detection of metallo-β-lactamase in imipenem-resistant Pseudomonas aeruginosa.
Carbapenems are the primary choice of treatment for severe Pseudomonas aeruginosa infection. However, the emergence of carbapenem resistance due to the production of metallo-β-lactamases (MBLs) is of global concern. In this study, 90 imipenem- (IPM- or IP-) resistant P. aeruginosa (IRPA) isolates, including 32 previously tested positive and genotyped for MBL genes by PCR, were subjected to double-disk synergy test (DDST), combined disk test (CDT), and imipenem/imipenem-inhibitor (IP/IPI) E-test to evaluate their MBLs detection capability. All three methods were shown to have a sensitivity of 100%. However, DDST was the most specific of the three (96.6%), followed by IP/IPI E-test interpreted based on the single criteria of IP/IPI ≥8 as positive (62.1%), and CDT was the least specific (43.1%). Based on the data from this evaluation, we propose that only IRPA with IP MIC >16 μg/mL and IP/IPI ≥8 by IP/IPI E-test should be taken as positive for MBL activity. With the new dual interpretation criteria, the MBL IP/IPI E-test was shown to achieve 100% sensitivity as well as specificity for the IRPA in this study. Therefore, the IP/IPI E-test is a viable alternative phenotypic assay to detect MBL production in IRPA in our population in circumstances where PCR detection is not a feasible option.


Do risk factors for problem behaviour act in a cumulative manner? An examination of ethnic minority and majority children through an ecological perspective.
Extensive research has identified risk factors for problem behaviour in childhood. However, most of this research has focused on isolated variables, ignoring possible additive influences. The purpose of this study was to examine whether risk factors for problem behaviour act in a cumulative manner, and to investigate whether cumulative risk stemming from distinct ecological levels (Bronfenbrenner, 1979) differentially influences the manifestation of problem behaviours in middle childhood. In addition, ethnic differences between minority (i.e., Indian) and majority (i.e., English) families were examined. The sample consisted of 125 children (59 English and 66 of Indian origin) between the ages of 7 and 9.6 (M = 8.51, SD = 0.62) and their parents. Both mothers and fathers completed questionnaires regarding the children's problem behaviour and provided reports of the children's characteristics and environment. Children were also assessed and provided reports about themselves and their relationships. Finally, parent-child mutuality and parenting behaviour were coded from a videotaped parent-child interaction task. Risk factors acted in a cumulative manner - the more risk children experienced, the more problem behaviour they exhibited. Total problem behaviour was predicted by all three levels: individual, microsystem and exosystem. However, externalising problems were mainly predicted by microsystem-level cumulative risk, whereas internalising problems were predicted by both individual-level cumulative risk and exosystem-level cumulative risk. These results were similar for both ethnic groups. The support for the cumulative hypothesis highlights the importance of having a broad picture of children's characteristics and environmental components when analysing children's adjustment. The distinct influence of risk stemming from the different ecological levels suggests that the trajectories of internalising, externalising and total problem behaviour may be different.


Internet-Delivered Versus Face-to-Face Cognitive Behavior Therapy for Anxiety Disorders: Systematic Review and Meta-Analysis.
Over the last 20 years, internet-delivered cognitive behavior therapy (ICBT) has been tested in a large number of randomized controlled trials, often with positive results. However, it is not widely known about the efficacy of ICBT as compared to face-to-face cognitive behavior therapy (CBT). In the present systematic review and meta-analysis, ICBT for treatment of anxiety disorders was directly compared to face- to-face CBT within the same trial. This study aimed to reinvestigate the effect of ICBT compared to face-to-face CBT for anxiety disorders. A total of 8 studies out of the 236 articles screened met all the inclusion criteria. The included studies targeting five different anxiety disorders, social anxiety disorder, adolescent anxiety, panic disorder, spider phobia, and fear of public speaking, had been carried out in Australia, Spain, and Sweden. The total number of participants was 348 in ICBT and 316 in face-to-face conditions. The results of our meta-analysis are interesting both from theoretical and practical standpoints, which showed a pooled effect size posttreatment with Hedges' g = 0.01 (95% CI: -0.16 to 0.18). ICBT and face-to-face CBT created equivalent overall effects. in treatment of anxiety disorders. Since there have been similar systematic reviews about anxiety disorders so far, and in majority of them, ICBT has not been compared against face-to-face treatment. More research is needed to establish the general equivalence of the two treatment formats. Also, understanding what makes ICBT work is a challenge for future research.


[Studies of ceftizoxime in perinatal period].
Bacteriological studies and clinical evaluations of ceftizoxime (CZX) in perinatal period were carried out, and results are summarised as follows: Antibacterial activities of CZX in amniotic fluid were determined using the broth dilution method, and bactericidal effect on Streptococcus agalactiae, Staphylococcus aureus and Escherichia coli were demonstrated. The bactericidal effect of CZX increased in amniotic fluid and remarkable increases of activities against resistant strains were demonstrated. The penetration of CZX into mother's milk was low, and it was speculated that the drug transfer to the newborn through breast feeding was very little. Clinically, CZX was effective in the treatment of perinatal infections without any side effect. The above results has demonstrated that CZX is a clinically useful antibiotic for the prophylaxis and the treatment of perinatal infections.


Osteoarticular brucellosis in children.
The findings in 40 children (24 female, 16 male) with osteoarticular complications of brucellosis are presented. Raw milk was the main source of infection. Most patients had acute onset of fever, arthralgia, and myalgia. Arthritis was the presenting symptom in all patients, of whom two also had osteomyelitis. Arthritis was monoarticular in 28 (70%) of 40 cases with predilection for large weight-bearing joints. Spine and small joints were spared. Brucella melitensis was detected in 23/31 (74%) cases. Mild anaemia, leucopenia, increased liver enzymes, positive acute phase reactants, and low titres of autoantibodies were prominent non-specific laboratory findings. Bone scintigraphy was more helpful than conventional radiography in detecting hip and sacroiliac joint disease. Treatment with a combination of antibiotics for six weeks or more resulted in a cure rate of 92.5%. Early recognition of infection, prolonged treatment, and long term follow up should improve the outcome of patients.


Hormone-induced rise in cytosolic Ca2+ in axolotl hepatocytes: properties of the Ca2+ influx channel.
Calcium entry in nonexcitable cells occurs through Ca(2+)-selective channels activated secondarily to store depletion and/or through receptor- or second messenger-operated channels. In amphibian liver, hormones that stimulate the production of adenosine 3',5'-cyclic monophosphate (cAMP) also regulate the opening of an ion gate in the plasma membrane, which allows a noncapacitative inflow of Ca2+. To characterize this Ca2+ channel, we studied the effects of inhibitors of voltage-dependent Ca2+ channels and of nonselective cation channels on 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP)-dependent Ca2+ entry in single axolotl hepatocytes. Ca2+ entry provoked by 8-BrcAMP in the presence of physiological Ca2+ followed first-order kinetics (apparent Michaelis constant = 43 microM at the cell surface). Maximal values of cytosolic Ca2+ (increment approximately 300%) were reached within 15 s, and the effect was transient (half time of 56 s). We report a strong inhibition of cAMP-dependent Ca2+ entry by nifedipine [half-maximal inhibitory concentration (IC50) = 0.8 microM], by verapamil (IC50 = 22 microM), and by SK&F-96365 (IC50 = 1.8 microM). Depolarizing concentrations of K+ were without effect. Gadolinium and the anti-inflammatory compound niflumate, both inhibitors of nonselective cation channels, suppressed Ca2+ influx. This "profile" indicates a novel mechanism of Ca2+ entry in nonexcitable cells.


Radiological Assessment of Paediatric Fungal Infections: A Pictorial Review With Focus on PET/MRI.
Paediatric invasive fungal infections have significantly increased over the past few decades, in particular among the immunocompromised population. Candida and Aspergillus spp. are still the most commonly isolated organisms. Image-based assessment of fungal infections can indeed be challenging especially in oncological patients where the differential diagnosis relative to other infections and neoplastic lesions cannot be often obvious. Therefore, the knowledge of the main radiological features associated with fungal infections is crucial to achieve an early correct diagnosis and address the most appropriate therapeutic approach. Thus, our aim was to review the main radiological features of paediatric fungal infections with particular focus on positron emission tomography/magnetic resonance imaging (PET/MRI), referring to the experience of our tertiary level hospital.


An estimate of the burden of serious fungal diseases in Greece.
Data on the epidemiology of serious fungal infections in Greece are scarce. Our aim was to calculate the burden of serious fungal diseases in Greece. A thorough literature search for papers reporting epidemiological data on serious fungal diseases in Greece was performed. Where no Greek data existed, we used a structured set of assumptions to estimate fungal disease burden, based on specific high-risk populations. Of the 10.8 million population, 85.5 % are adults and 27 % are over 60 years of age. The annual fungal disease estimates are as follows: 142,337 Greek women get recurrent vaginal thrush (2,632 cases/100,000 females); there are 889 cases of esophageal candidiasis (8.2 cases/100,000); annual incidence of Pneumocystis pneumonia is 112 cases; chronic pulmonary aspergillosis prevalence is 386 cases; there are 20,843 patients with allergic bronchopulmonary aspergillosis and 27,744 with severe asthma with fungal sensitization; candidaemia incidence is 541 cases (5.0/100,000); there are 81 cases of Candida peritonitis; invasive aspergillosis occurs in 1,125 patients. According to our calculations, 194,067 individuals (1.79 cases/100,000) in Greece suffer from serious fungal diseases each year. This is the first attempt to determine the burden of fungal diseases in Greece, and provides a crude estimate on its impact on public health.


Initial radiographic features as outcome predictor of adult respiratory syncytial virus respiratory tract infection.
The purposes of this study were to evaluate the chest radiographic features of adult patients hospitalized for respiratory syncytial virus respiratory tract infections and to assess whether initial chest radiographic findings help predict clinical outcome. All adult patients hospitalized from January 2009 to December 2011 with laboratory-confirmed respiratory syncytial virus infection were included in the study. Patient clinical data and admission chest radiographs were retrospectively reviewed. Adverse outcomes included need for supplemental oxygen, need for assisted ventilation, and death. Of 285 patients (mean age, 74 ± 16 years) included, 199 (69.8%) had abnormal chest radiographic findings: 49.5% (141/285) had acute changes, and 47.7% (136/285) had chronic changes. Consolidation (68/141 [48.2%]) and ground-glass opacity (57/141 [40.4%]) were the predominant types of acute changes and were most common in unilateral single-lower-zone involvement. Consolidation, ground-glass opacity, and chronic changes occurred with significantly higher frequency in patients with adverse outcomes. The presence of acute (odds ratio, 3.6) and chronic (odds ratio, 2.2) changes were independent risk factors for mortality. A large proportion of adult patients hospitalized with respiratory syncytial virus respiratory tract infection had changes on initial chest radiographs. Consolidation or ground-glass opacity in a unilateral single-lower-zone distribution were the most common findings. The presence of acute and chronic radiographic lung changes was associated with adverse outcomes.


EUS-guided biliary drainage: A systematic review and meta-analysis.
ERCP is the current procedure of choice for patients with jaundice caused by biliary obstruction. EUS-guided biliary drainage (EUS-BD) has emerged as an alternative to ERCP in patients requiring biliary drainage. The aim of the study was to conduct a systematic review and meta-analysis to report the overall efficacy and safety of EUS-BD. We conducted a comprehensive search of several databases including PubMed, EMBASE, Web of Science, Google Scholar, and LILACS databases (earliest inception to June 2018) to identify studies that reported EUS-BD in patients. The primary outcome was to look at the technical and clinical success of the procedure. The secondary analysis focused on calculating the pooled rate of re-interventions and all adverse-events, along with the commonly reported adverse-event subtypes. Twenty-three studies reporting on 1437 patients were identified undergoing 1444 procedures. Majority of the patient population were male (53.86%), with an average age of 67.22 years. The pooled technical success rates and clinical success rates were 91.5% (95% confidence interval [CI]: 87.7-94.2, I[2] = 76.5) and 87% (95% CI: 82.3-90.6, I[2] = 72.4), respectively. The total adverse event rates were 17.9% (95% CI: 14.3-22.2, I[2] = 69.1). Subgroup analysis of three major individual adverse events was bile leak: 4.1% (2.7-6.2, I[2] = 46.7), stent migration: 3.9% (2.5-6.2, I[2] = 43.5), and infection: 3.8% (2.8-5.1, I[2] = 0) Substantial heterogeneity was noted in the analysis. EUS-BD has high technical and clinical success rate and hence a very effective procedure. Concerns about publication bias exist. Careful consideration should be given to the adverse events and weighing the risks and benefits of the alternative nonsurgical/surgical approaches.


Vav1 and mutant K-Ras synergize in the early development of pancreatic ductal adenocarcinoma in mice.
To explore the contribution of Vav1, a hematopoietic signal transducer, to pancreatic ductal adenocarcinoma (PDAC) development, we generated transgenic mouse lines expressing, Vav1, K-RasG12D, or both K-RasG12D and Vav1 in pancreatic acinar cells. Co-expression of Vav1 and K-RasG12D synergistically enhanced acinar-to-ductal metaplasia (ADM) formation, far exceeding the number of lesions developed in K-RasG12D mice. Mice expressing only Vav1 did not develop ADM. Moreover, the incidence of PDAC in K-RasG12D/Vav1 was significantly higher than in K-RasG12D mice. Discontinuing Vav1 expression in K-RasG12D/Vav1 mice elicited a marked regression of malignant lesions in the pancreas, demonstrating Vav1 is required for generation and maintenance of ADM. Rac1-GTP levels in the K-RasG12D/Vav1 mice pancreas clearly demonstrated an increase in Rac1 activity. Treatment of K-RasG12D and K-RasG12D/Vav1 mice with azathioprine, an immune-suppressor drug which inhibits Vav1's activity as a GDP/GTP exchange factor, dramatically reduced the number of malignant lesions. These results suggest that Vav1 plays a role in the development of PDAC when co-expressed with K-RasG12D via its activity as a GEF for Rac1GTPase.


Development and validation of the Generalized Anxiety Disorder Inventory (GADI).
The psychometric tools used for the assessment of generalized anxiety disorder (GAD) either do not conform to the current concept of the condition or have important limitations. We aimed to develop and validate a new questionnaire for the assessment of symptom profile and severity of GAD. An original pool of potential scale items was subjected to a series of studies in non-clinical and clinical populations, in order to determine the final composition of the scale. The psychometric properties of the new scale, the Generalized Anxiety Disorder Inventory (GADI), were evaluated using a factor analytic model suitable for ordinal data and the Graded Response Model. The precision of measurement of the GADI was quantified through the item information functions.A total of 197 outpatients and 522 non-clinical subjects participated in four studies and completed the GADI. The final 18-item scale was derived from an original pool of 30 potential items. The GADI showed good reliability, convergent and divergent validity. The scale comprises three factors, relating to cognitive, somatic and sleep symptoms. It accurately distinguished GAD patients from non-patient controls. The cognitive factor also distinguished GAD from other anxiety disorders and depression. The GADI is a useful tool in the assessment of the breadth of symptoms and the severity of generalized anxiety disorder in clinical settings.


Surgical resection plus adjuvant radiotherapy is superior to surgery or radiotherapy alone in the prevention of neurological decline in a rat metastatic spinal tumor model.
The optimal management of spinal column metastatic disease is controversial. Furthermore, the literature lacks an accurate animal model to study the efficacy of surgical treatment options for spinal column metastases. We compared the efficacy of surgery, radiotherapy, or surgery plus adjuvant radiotherapy in a rat model of metastatic epidural spinal cord compression. Thirty-two Fischer 344 rats underwent a transabdominal approach for implantation of a CRL-1666 breast adenocarcinoma cell line within the vertebral body of L6. Animals were randomly assigned to receive one of four treatments (n = 8 per group) 7 days after tumor implantation: 1) control: no treatment; 2) external beam radiation therapy (XRT) (total 20 Gy in 400-cGy daily fractions); 3) surgery: L6 vertebral corpectomy, tumor resection, and polymethyl methacrylate reconstruction; and 4) surgery + XRT: corpectomy and tumor resection followed by XRT (total 20 Gy in 400-cGy daily fractions) 72 hours after surgery. Hind-limb function was tested daily after treatment using the Basso-Beattie-Bresnahan (BBB) scale (range, 1-21). All animals (n = 32) demonstrated normal hind-limb function (BBB score, 21) on posttreatment Day 1. The XRT, surgery, and surgery + XRT groups all experienced a delay in onset of paresis versus the control group. Compared to the XRT group, the surgery group demonstrated greater median BBB scores on Days 3 (21 versus 20, P = 0.02) through 9 (12 versus 8, P = 0.002) after treatment. Compared with the surgery group, the surgery + XRT group demonstrated even greater median BBB scores on Days 6 (21 versus 19, P = 0.0008) through 11 (16 versus 8, P = 0.0001) after treatment. Median time to loss of ambulation (BBB <or= 7) was greatest in the surgery + XRT group (15 d) when compared with the surgery (12 d, P = 0.001), XRT (9 d, P = 0.001), or control groups (7 d, P = 0.0005). In a rat model of metastatic epidural spinal cord compression, decompressive surgery followed by radiotherapy yielded the greatest efficacy in the prevention of neurological decline when compared with surgery or radiotherapy alone. Radiotherapy alone attenuated neurological decline but was the least efficacious treatment in this model. These results support this animal model as an effective platform to investigate novel interventions for metastatic spine tumors.


Transgene excision in zebrafish using the phiC31 integrase.
Genome engineering strategies employing site-specific recombinases (SSRs) have become invaluable to the study of gene function in model organisms. One such SSR, the integrase encoded by the Streptomyces bacteriophage phiC31, promotes recombination between heterotypic attP and attB sites. In the present study I have examined the feasibility of the use of phiC31 integrase for intramolecular recombination strategies in zebrafish embryos. I report here that (1) phiC31 integrase is functional in zebrafish cells, (2) phiC31 integrase can excise a transgene cassette flanked by an attB and an attP site, analogous to a common use of the Cre/lox SSR system, (3) phiC31 integrase functions in the zebrafish germline, and (4) a phiC31 integrase-estrogen receptor hormone-binding domain variant fusion protein catalyzes attB-attP recombination in zebrafish embryos in a 4-hydroxytamoxifen-dependent manner, albeit less efficiently than phiC31 alone. These features should make this a useful approach for genome manipulations in the zebrafish.


The origin and evolution of mutations in acute myeloid leukemia.
Most mutations in cancer genomes are thought to be acquired after the initiating event, which may cause genomic instability and drive clonal evolution. However, for acute myeloid leukemia (AML), normal karyotypes are common, and genomic instability is unusual. To better understand clonal evolution in AML, we sequenced the genomes of M3-AML samples with a known initiating event (PML-RARA) versus the genomes of normal karyotype M1-AML samples and the exomes of hematopoietic stem/progenitor cells (HSPCs) from healthy people. Collectively, the data suggest that most of the mutations found in AML genomes are actually random events that occurred in HSPCs before they acquired the initiating mutation; the mutational history of that cell is "captured" as the clone expands. In many cases, only one or two additional, cooperating mutations are needed to generate the malignant founding clone. Cells from the founding clone can acquire additional cooperating mutations, yielding subclones that can contribute to disease progression and/or relapse.


Progression in multiple sclerosis is associated with low endogenous NCAM.
Multiple sclerosis (MS) is a CNS disorder characterized by demyelination and neurodegeneration. Although hallmarks of recovery (remyelination and repair) have been documented in early MS, the regenerative capacity of the adult CNS per se remains uncertain with the wide held belief that it is either limited or non-existent. The neural cell adhesion molecule (NCAM) is a cell adhesion molecule that has been widely implicated in axonal outgrowth, guidance and fasciculation. Here, we used in vitro and in vivo of MS to investigate the role of NCAM in disease progression. We show that in health NCAM levels decrease over time, but this occurs acutely after demyelination and remains reduced in chronic disease. Our findings suggest that depletion of NCAM is one of the factors associated with or possibly responsible for disease progression in MS.


GPR Expression in Intestinal Biopsies From SCT Patients Is Upregulated in GvHD and Is Suppressed by Broad-Spectrum Antibiotics.
Microbiota can exert immunomodulatory effects by short-chain fatty acids (SCFA) in experimental models of graft-versus-host disease (GvHD) after allogeneic hematopoietic stem cell transplantation (allo-SCT). Therefore we aimed to analyze the expression of SCFAs sensing G-protein coupled receptor GPR109A and GPR43 by quantitative PCR in 338 gastrointestinal (GI) biopsies obtained from 199 adult patients undergoing allo-SCT and assessed the interaction of GPR with FOXP3 expression and regulatory T cell infiltrates. GPR expression was strongly upregulated in patients with stage II-IV GvHD (p=0.000 for GPR109A, p=0.01 for GPR43) and at the onset of GvHD (p 0.000 for GPR109A, p=0.006 for GPR43) and correlated strongly with FOXP3 and NLRP3 expression. The use of broad-spectrum antibiotics (Abx) drastically suppressed GPR expression as well as FOXP3 expression in patients' gut biopsies (p=0.000 for GPRs, FOXP3 mRNA and FOXP3+ cellular infiltrates). Logistic regression analysis revealed treatment with Abx as an independent factor associated with GPR and FOXP3 loss. The upregulation of GPRs was evident only in the absence of Abx (p=0.001 for GPR109A, p=0.014 for GPR43) at GvHD onset. Thus, GPR expression seems to be upregulated in the presence of commensal bacteria and associates with infiltration of FOXP3+ T regs, suggesting a protective, regenerative immunomodulatory response. However, Abx, which has been shown to induce dysbiosis, interferes with this protective response.


Intestinal dysplasia and adenoma in transgenic mice after overexpression of an activated beta-catenin.
Mutations in the adenomatous polyposis coli gene or activating mutations in the beta-catenin gene itself are thought to be responsible for the excessive beta-catenin signaling involved in intestinal carcinogenesis. We generated transgenic mice that expressed large amounts of a NH2-terminally truncated mutant beta-catenin (deltaN131beta-catenin) in the intestine. These mice had multifocal dysplastic lesions in the small intestine, reminiscent of the early lesions observed in the mouse models of familial adenomatous polyposis. The number of apoptotic cells in the villi of these transgenic mice was 3-4-fold higher than in nontransgenic mice. Expression of the truncated beta-catenin mutant in the kidney led to the development of severe polycystic kidney disease. Our findings support the concept that deregulation of the beta-catenin signaling pathway is the major oncogenic consequence of adenomatous polyposis coli mutations in intestinal neoplasia.


MOPC 104E plasmacytoma functional heterogeneity and maturational potential in culture.
MOPC 104E plasmacytoma secretes an immunoglobulin M (IgM) paraprotein which reacts specifically with dextran B-1355. We separated plasmacytoma cells into subfractions by density gradient centrifugation. The majority of cells isolated from the peritoneal washes 3 days following i.p. transplantation of tumor were in active cell cycle and had a density of 1.055 or 1.065 g/ml. Cells isolated from 9-day-old ascites were confined to the heavy-density fraction (1.085 g/ml). The majority of these cells were not in proliferative phase. Functional analysis of the fractions using rosette formation and plaque formation assays indicated that cells in active cell cycle had less surface IgM and secreted less IgM than did cells not in cell cycle. An attempt was made to establish a cultured cell line of MOPC 104E plasmacytoma. The majority of the cells of the continuously cultured cell line were in active cell cycle, had less surface IgM, and secreted less IgM. Cultured cells acquired more cell surface IgM and actively secreted IgM following their secondary colonization in 0.8% methylcellulose. These studies showed that the new line of MOPC 104E retained the properties of the original ascites tumor. An important feature of these studies is that the density of cells within a colony is a stable property and is probably not related to cell cycle.


Infection prevention and control in the operating theatre: reducing the risk of surgical site infections (SSIs).
Continuing advances in surgical techniques, asepsis, operating theatre protocols and ventilation systems that ensure an uninterrupted supply of clean air, should allow all patients to undergo both invasive and minimally-invasive procedures with reduced risk. Patients having surgery in the United Kingdom are probably less vulnerable to surgical site infections (SSIs) than ever before--despite persisting concerns about meticillin-resistant Staphylococcus aureus (MRSA) and increasing antibiotic resistance in other organisms such as vancomycin-resistant Enterococci (VRE).


My method of extracapsular cataract extraction with implantation of a posterior chamber intraocular lens.
Simplified manual forms of extracapsular cataract extraction appear to be more economical, yet as safe and simple as techniques employing more complex instrumentation. The following method, employed in 40,000 cases, seems to the author to be an efficient and effective way of performing cataract extraction. One hour prior to surgery, modified retrobulbar anesthesia alone is employed using the technique described, and provides adequate akinesia as well as anesthesia. Positive pressure is applied to the eye preoperatively to assure softness of the globe at the time of surgery. An anterior capsulectomy is made with scissors, using the open-sky technique. Following the delivery of the lens nucleus and as much cortex as possible with a lens loop, the remaining cortex is removed with an angled, 23-gauge, double irrigation-aspiration cannula. Filtered balance salt solution, containing gentamycin flows into the eye through the side port (inflow) and a 3-cc syringe with 2 cc BSS for irrigation-aspiration is connected to the other (aspiration) port. A single, 25-gauge, angled irrigation cannula is used to clean up the finger cortical remains. A firm-loop intraocular lens is inserted into the capsular bag. The posterior capsule is left intact.


Recombinant versus urinary human chorionic gonadotrophin for final oocyte maturation triggering in IVF and ICSI cycles.
For the last few decades urinary human chorionic gonadotrophin (uhCG) has been used to trigger final oocyte maturation in cycles of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). Recombinant technology has allowed the production of two drugs, recombinant human chorionic gonadotrophin (rhCG) and recombinant luteinising hormone (rLH), that can be used for the same purpose, to mimic the endogenous luteinising hormone (LH) surge. This allows commercial manufacturers to adjust production according to market requirements and to remove all urinary contaminants, facilitating the safe subcutaneous administration of a compound with less batch-to-batch variation. However, prior to a change in practice, it is necessary to compare the effectiveness of the recombinant drugs to the currently used urinary human chorionic gonadotrophin (uhCG). To assess the effects of subcutaneous rhCG and high dose rLH versus uhCG for inducing final oocyte maturation in subfertile women undergoing IVF and ICSI cycles. We searched the Cochrane Menstrual Disorders and Subfertility Group Trials Register (April 2015), the Cochrane Central Register of Controlled Trials (CENTRAL) (2015, Issue 3), MEDLINE (1946 to April 2015), EMBASE (1980 to April 2015) and PsycINFO (1806 to April 2015) as well as trial registers at ClinicalTrials.gov on 13 May 2015 and the World Health Organization International Clinical Trials Registry Platform (WHO ICTRP) search portal on 14 May 2015. Two review authors independently scanned titles and abstracts and selected those that appeared relevant for collection of the full paper. We included randomised controlled trials comparing rhCG and rLH with urinary hCG for final oocyte maturation triggering in IVF and ICSI cycles for treatment of infertility in normogonadotropic women. Two authors independently performed assessment for inclusion or exclusion, quality assessment and data extraction. We discussed any discrepancies in the presence of a third author to reach a consensus. The primary review outcomes were ongoing pregnancy/live birth and incidence of ovarian hyperstimulation syndrome (OHSS). Clinical pregnancy, miscarriage rate, number of oocytes retrieved and adverse events were secondary outcomes. We combined data to calculate pooled odds ratios (ORs) and 95% confidence intervals (CIs) and assessed statistical heterogeneity using the I(2) statistic. We evaluated the overall quality of the evidence for the main comparisons using GRADE methods. We included 18 RCTs involving 2952 participants; 15 compared rhCG with uhCG, and 3 compared rhLH with uhCG. The evidence for different comparisons ranged from very low to high quality: limitations were poor reporting of study methods and imprecision. Pharmaceutical companies funded 9 of the 18 studies, and 5 studies did not clearly report funding source. Ongoing pregnancy/live birthThere was no conclusive evidence of a difference between rhCG and uhCG (OR 1.15, 95% CI 0.89 to 1.49; 7 RCTs, N = 1136, I(2) = 0%, moderate quality evidence) or between rhLH and uhCG (OR 0.95, 95% CI 0.51 to 1.78, 2 RCTs, N = 289, I(2) = 0%, very low quality evidence) for ongoing pregnancy/live birth rates. OHSS There was no evidence of a difference between rhCG and uhCG in the incidence of OHSS: moderate to severe OHSS (OR 1.76, 95% CI 0.37 to 8.45; 3 RCTs, N = 417, I(2) = 0%, low quality evidence), moderate OHSS (OR 0.78, 95% CI 0.27 to 2.27; 1 RCT, N = 243, I(2) = 0%, low quality evidence), mild to moderate OHSS (OR 1.00, 95% CI 0.42 to 2.38; 2 RCTs, N = 320, I(2) = 0%, low quality evidence) or undefined OHSS (OR 1.18, 95% CI 0.50 to 2.78; 3 RCTs, N = 495, I(2) = 0%, low quality evidence). Likewise, there was no evidence of a difference between rhLH and uhCG in OHSS rates for moderate OHSS (OR 0.82, 95% CI 0.39 to 1.69, 2 RCTs, N = 280, I(2) = 5%, very low quality evidence). Other adverse events There was no evidence of a difference in miscarriage rates between rhCG and uhCG (OR 0.72, 95% CI 0.41 to 1.25; 8 RCTs, N = 1196, I(2) = 0%, low quality evidence) or between rhLH and uhCG (OR 0.95, 95% CI 0.38 to 2.40; 2 RCTs, N = 289, I(2) = 0%, very low quality evidence). For other adverse effects (most commonly injection-site reactions) rhCG was associated with a lower number of adverse events than uhCG (OR 0.52, 95% CI 0.35 to 0.76; 5 RCTS, N = 561; I(2) = 67%, moderate quality evidence). However, when we used a random-effects model due to substantial statistical heterogeneity, there was no evidence of a difference between the groups (OR 0.56, 95% CI 0.27 to 1.13). Only one study comparing rLH and uhCG reported other adverse events, and it was impossible to draw conclusions. We conclude that there is no evidence of a difference between rhCG or rhLH and uhCG for live birth or ongoing pregnancy rates or rates of OHSS.


Osseointegration around titanium screws placed into the areas between guided bone augmented sites compared with osseointegration around guided bone graft augmented sites in rabbit tibia.
The aim of this study was to compare the degree of osseointegration around titanium screws placed into the areas between guided bone augmented sites and guided bone graft augmented sites in 8 Japanese white rabbit tibia. The skin and periosteum were incised and lifted to expose the inner surfaces of both tibia. Eight rabbits were divided into 2 groups: 1) a guided bone augmentation (GBA) group of 2 rabbits, and 2) a guided bone graft augmentation (GBCA) group of 6 rabbits. In the GBA group, the bone marrow was penetrated several times in both tibial bones. Three titanium screws were inserted into the bone so that the top of each screw was 3 mm above the bone surface and covered with an expanded polytetrafluoroethylene membrane (ePTFE). In the GBGA group autogenous cortical bone (10 mm x 10 mm x 1 mm) was taken from the left tibia and grafted on the inner surface of the right tibial bone, titanium screws being similarly inserted. The surgical site was covered by ePTFE. After healing for 2 months, the animals were euthanatized, and the experimental area was prepared for histological investigation. New bone had formed on the surgical sites in the GBA group and grafted bone had survived in the GBGA group, with no sign of infection or membrane exposure. The degree of osseointegration around the screw was 67.6 +/- 11.2% (mean +/- SD) in the GBA group and 78.8 +/- 11.9% in the GBGA group. There was no significant difference between the groups (p = 0.29). The ratio of mineralized bone to bone marrow was 45.7 +/- 6.2% in the GBA group and 64.4 +/- 4.4% in the GBGA group. There was a significant difference between the groups (p < 0.05). Although there was no significant difference for osseointegration around the screw placed into the GBA and GBGA sites, the quality of newly generated bone or grafted bone was quite different. Therefore, we suggest that predictable bone augmentation beyond the bone surface can be achieved by either the GBA or the GBGA procedure with ePTFE. However, it was difficult to conclude which procedure was superior with respect to the osseointegration around the titanium screw.


Seroprevalence of Toxoplasma gondii in household and stray cats of Riyadh, Saudi Arabia.
This study was undertaken to determine the seroprevalence of Toxoplasma gondii in cats in the area of Riyadh, Saudi Arabia. We examined 200 serum samples collected from stray and household cats for T. gondii antibodies by ELISA. The overall seroprevalence was 26%. Seroprevalence was significantly higher (p < 0.05) in stray cats (39%) compared with household cats (13%). The prevalence in male and female cats was 31.4% and 20.4%, respectively. The seroprevalence increased with age and was higher in cats over 6 years of age (43%) as opposed to cats less than 4 years old (33%). Seropositivity varied according to the breed. The highest was recorded among cats of American breed (38.5%), followed by Persian (27%), Himalayan (21%), Bengali (11.5%), and Siamese (2%). Antibodies were not reported from the Turkish breed. Overall seroprevalence among cats did not vary significantly with season or with the localities within the Riyadh municipality. We also examined 100 faecal samples from stray and household cats by flotation technique, which revealed an overall prevalence of 12% of T. gondii oocycts.


[Atrial septostomy in pulmonary hypertension].
Despite the fact that remarkable improvements have been achieved in quality of life and survival through increasing number of specific medical treatment alternatives in severe pulmonary hypertension, many patients have resistance to medical treatment. Atrial septostomy is considered as a palliative procedure in severe pulmonary hypertension to relieve symptoms of severe pulmonary hypertension by reducing right ventricular preload and increasing systemic flow. Gradual balloon dilatation and blade balloon atrial septostomy are the most frequently used methods. If atrial septostomy is performed in experienced medical centers, it provides clinical improvement and even has tendency to extend life expectancy in patients with pulmonary hypertension resistant to medical treatment.


On pose recovery for generalized visual sensors.
With the advances in imaging technologies for robot or machine vision, new imaging devices are being developed for robot navigation or image-based rendering. However, to satisfy some design criterion, such as image resolution or viewing ranges, these devices are not necessarily being designed to follow the perspective rule and, thus, the imaging rays may not pass through a common point. Such generalized imaging devices may not be perspective and, therefore, their poses cannot be estimated with traditional techniques. In this paper, we propose a systematic method for pose estimation of such a generalized imaging device. We formulate it as a nonperspective n point (NPnP) problem. The case with exact solutions, n=3, is investigated comprehensively. Approximate solutions can be found for n>3 in a least-squared-error manner by combining an initial-pose-estimation procedure and an orthogonally iterative procedure. This proposed method can be applied not only to nonperspective imaging devices but also perspective ones. Results from experiments show that our approach can solve the NPnP problem accurately.


Folic acid and colorectal cancer prevention: molecular mechanisms and epidemiological evidence (Review).
Epidemiological studies suggest that a diet rich in fruits and vegetables protects against colorectal cancer. This effect may result from their high levels of folic acid (pteroylglutamic acid). Folic acid holds a key position in DNA synthesis and mitosis as well as DNA methylation and regulation of gene expression. Folic acid deficiency has been associated with site- and gene specific DNA hypo- and hypermethylation. Furthermore thymidylate synthesis is restricted by folic acid deficiency which causes misincorporation of nucleotides and DNA strand breaks. Much epidemiological evidence supports the hypothesis that insufficient folic acid supply favors the development of colorectal tumors, particularly prospective studies have supported this connection. However, the data from case-control studies are less consistent. Functional polymorphisms in folate-metabolizing genes, especially the methylenetetrahydrofolate reductase (MTHFR) are capable of modifying the risk of colorectal cancer. Observational studies show that individuals with the homozygote genotype for the MTHFR (677C-->T) polymorphism are at higher risk when folic acid supply is low. Currently there are only few human intervention trials which show that folic acid can modify and inhibit the development of colorectal tumors. Additional studies are required in order to determine whether folic acid will be a useful agent in colorectal cancer prevention.


Evaluation of a hemagglutination test for human leptospirosis.
An indirect hemagglutination test for the diagnosis of leptospirosis is described; the test uses a soluble antigen from serotype patoc to sensitize sheep erythrocytes which are then fixed with glutaraldehyde. Evaluation of this procedure indicates that it is more reliable than the conventional macroscopic agglutination test and, in contrast with both microscopic and macroscopic agglutination tests, is positive only with sera from persons with current leptospiral illness. The test is simple and convenient and sensitized fixed cells may be stored for at least a year. In comparison with the macroscopic and microscopic tests, only a single antigen is required.


Dynamic adaptation of cardiac oxidative phosphorylation is not mediated by simple feedback control.
The classic idea about regulation of cardiac oxidative phosphorylation (OxPhos) was that breakdown products of ATP (ADP and P(i)) diffuse freely to the mitochondria to stimulate OxPhos. On the basis of this metabolic feedback control system, the response time of OxPhos (t(mito)) is predicted to be inversely proportional to the mitochondrial aerobic capacity (MAC). We determined t(mito) during steps in heart rate in isolated perfused rabbit hearts (n = 16) before and after reducing MAC with nonsaturating doses of oligomycin. The reduction of MAC was quantified in mitochondria isolated from each perfused heart, dividing oligomycin-sensitive, ADP-stimulated state 3 respiration by oligomycin-insensitive uncoupled respiration. The t(mito) to heart rate steps from 60 to 70 and 80 beats/min was 5. 6 +/- 0.6 and 7.2 +/- 0.8 s (means +/- SE) and increased an estimated 34 and 40% for a 50% decrease in MAC (P < 0.05), respectively, which is much less than the 100% predicted by the feedback hypothesis. For steps to 100 or 120 beats/min, t(mito) was 8.3 +/- 0.5 and 11.2 +/- 0.6 s and was not reduced with decreases in MAC (P > 0.05). We conclude that immediate feedback control by quickly diffusing ADP and P(i) cannot explain the dynamic regulation of cardiac OxPhos. Because calcium entry into the mitochondria also cannot explain the first fast phase of OxPhos activation, we propose that delay of the energy-related signal in the cytoplasm dominates the response time of OxPhos.


Tethered nanoparticle-polymer composites: phase stability and curvature.
Phase behavior of poly(ethylene glycol) (PEG) tethered silica nanoparticles dispersed in PEG hosts is investigated using small-angle X-ray scattering. Phase separation in dispersions of densely grafted nanoparticles is found to display strikingly different small-angle X-ray scattering signatures in comparison to phase-separated composites comprised of bare or sparsely grafted nanoparticles. A general diagram for the dispersion state and phase stability of polymer tethered nanoparticle-polymer composites incorporating results from this as well as various other contemporary studies is presented. We show that in the range of moderate to high grafting densities the dispersion state of nanoparticles in composites is largely insensitive to the grafting density of the tethered chains and chemistry of the polymer host. Instead, the ratio of the particle diameter to the size of the tethered chain and the ratio of the molecular weights of the host and tethered polymer chains (P/N) are shown to play a dominant role. Additionally, we find that well-functionalized nanoparticles form stable dispersions in their polymer host beyond the P/N limit that demarcates the wetting/dewetting transition in polymer brushes on flat substrates interacting with polymer melts. A general strategy for achieving uniform nanoparticle dispersion in polymers is proposed.


ChatGPT for phenotypes extraction: one model to rule them all?
Information Extraction (IE) is a core task in Natural Language Processing (NLP) where the objective is to identify factual knowledge in textual documents (often unstructured), and feed downstream use cases with the resulting output. In genomic medicine for instance, being able to extract the most precise list of phenotypes associated to a patient allows to improve genetic disease diagnostic, which represents a vital step in the modern deep phenotyping approach. As most of the phenotypic information lies in clinical reports, the challenge is to build an IE pipeline to automatically recognize phenotype concepts from free-text notes. A new machine learning paradigm around large language models (LLM) has given rise of an increasing number of academic works on this topic lately, where sophisticated combinations of different technics have been employed to improve the phenotypes extraction accuracy. Even more recently released, the ChatGPT1 application nevertheless raises the question of the relevance of these approches compared to this new generic one based on an instruction-oriented LLM. In this paper, we propose a rigorous evaluation of ChatGPT and the current state-of-the-art solutions on this specific task, and discuss the possible impacts and the technical evolutions to consider in the medical domain.Clinical relevance- Deep phenotyping on electronic health records has proven its ability to improve genetic diagnosis by clinical exomes [10]. Thus, comparing state-of-the-art solutions in order to derive insights and improving research paths is essential.


Hemodynamic effects of platelet-activating factor in nonpregnant and pregnant sheep.
The present study was designed to assess the dose-related effects of platelet-activating factor (PAF) on systemic, renal, and uterine hemodynamics in nonpregnant sheep and to evaluate how pregnancy might alter these responses. Nonpregnant and pregnant (110 +/- 5 days gestation) ewes were instrumented for conscious measurements of maternal mean arterial pressure (MAP), renal blood flow (RBF), uterine blood flow (UBF), hematocrit, and urinary protein concentration. After recovery, dose-response curves to PAF were generated by systemic infusion at 10, 30, and 100 ng. kg(-1). min(-1) (15 min/dose) into the maternal femoral vein. The above parameters were measured, and renal and uterine vascular resistances (RVR and UVR, respectively) were calculated. In pregnant sheep, PAF increased MAP, RVR, UVR, and urinary protein concentration. We also observed increases in hematocrit, indicative of reduced blood volume secondary to increased systemic microvascular protein permeability. These responses were similar in nonpregnant sheep, with the exception of UVR in nonpregnant ewes being decreased (and thus UBF was increased), whereas in pregnant sheep, UVR was increased, which resulted in decreased UBF. This suggests that pregnancy alters the mechanism of action of PAF within the uterine vasculature in a way that can reduce UBF and thereby potentially compromise placental perfusion.


The benefits of drinking water quality regulation--England and Wales.
This paper aims to demonstrate that the regulation of drinking water quality in England and Wales has been successful in securing the improvements to drinking water quality resulting in better performance against EU and national standards. The water industry in England and Wales went through a major change in 1989 when suppliers were privatised and the government set up a robust regulatory regime. The regime was necessary as the industry was, as a result of privatisation, a monopoly with customers having no choice of supplier, unlike what was later available with other utilities such as gas or electricity. The regime would protect the interests of the consumer, the environment and public health through the quality of the product. The Drinking Water Inspectorate (DWI), as established in 1990, had to ensure the implementation of the European Drinking Water Directive (DWD) that had been transposed into national legislation. The aim of the DWD is to ensure that all EU Member States provide drinking water of a prescribed quality. In England and Wales, a body was required to oversee the performance of the industry against those standards, reporting to the Government and the European Commission. Through acts and legislation, the set up of the industry, the duties of the suppliers and regulators and the powers available to the regulators were established. The improvements to drinking water quality since privatisation were achieved by massive investment of the privatised water industry overseen by an independent regulator with clear duties and the powers to inspect, enforce and prosecute. The DWI's achievements show that to improve quality performance with the ability to report in detail how the improvements were made with extensive data evidence, a special regulator is required. The DWI advises policy departments and Ministers and when there are serious concerns regarding a threat to human health through drinking water, the highest level of regulatory power is the creation of new legislation, for example, the Cryptosporidium regulations that are unique to the UK. The DWI is more than what is traditionally thought of as a regulator because it has a single remit--drinking water quality--and its style of regulation has been key to improved drinking water quality in England and Wales.


Radiation myelitis after hypofractionated radiotherapy with concomitant gefitinib.
We describe the case of a 71-year-old Caucasian female with primary disseminated non-small cell cancer of the lung, presented for palliative radiotherapy of metastatic spread to the 9th and 11th thoracic vertebrae without intramedullary growth. Palliative radiotherapy with daily fractions of 3 Gy and a cumulative dose of 36 Gy to thoracic vertebrae 8-12 was performed. The patient received concomitantly 250 mg gefitinib daily. After a latent period of 16 months, the patient developed symptoms of myelitis. Magnetic resonance imaging (MRI) did not reveal any bony or intraspinal tumor progression, but spinal cord signal alteration. No response to steroids was achieved. The neurological symptoms were progressive in August 2013 with the right leg being completely plegic. The left leg was incompletely paralyzed. Deep and superficial sensitivity was also diminished bilaterally. The patient was completely urinary and anally incontinent. Contrary to the clinical findings, a follow-up MRI (July 2013) showed amelioration of the former signal alterations in the spinal cord. The diagnosis of paraneoplastic myelopathy was refuted by a negative test for autologous antibodies. At the last clinical visit in May 2014, the neurological symptoms were stable. The last tumor-specific treatment the patient is receiving is erlotinib 125 mg/d.We reviewed the literature and found no reported cases of radiation myelopathy after the treatment in such a setting. The calculated probability of such complication after radiotherapy alone is statistically measurable at the level of 0.02%. We suppose that gefitinib could also play a role in the development of this rare complication.


Cranialization of the frontal sinus--the final remedy for refractory chronic frontal sinusitis.
Chronic sinusitis can be a debilitating disease with significant impact on quality of life. Frontal sinusitis has a relatively low prevalence, but complications can be severe due to its anatomical location. After failure of conservative measures, typically endoscopic procedures are performed to improve the drainage of the frontal sinus. The cranialization of the frontal sinus is the final surgical measure, in which the affected frontal sinus is truly removed. In this study the authors describe the surgical technique of cranialization of the frontal sinus for refractory chronic frontal sinusitis, systematically search the literature for its application, and assess patient satisfaction in a cohort of consecutively treated patients after long-term follow-up. A consecutive cohort of 15 patients with refractory chronic frontal sinusitis was treated by cranialization of the frontal sinus and followed over a 20-year period (1989-2008) for the direct results and complications of the surgery. Long-term follow-up (mean 6.5 years) was obtained to assess the long-term effects of the cranialization. In all patients the signs and symptoms of chronic frontal sinusitis responded very well to the cranialization. Five patients had surgical complications, of which 2 were serious. One patient died of an unrelated cause and 1 patient was lost to follow-up. The remaining 13 patients had a long-term follow-up, which revealed that 12 of them thought that their life was better after the surgical procedure. Cranialization of the frontal sinus deserves consideration as the final remedy for refractory chronic frontal sinusitis after definite failure of other options.


Electroacupuncture Inhibits Neuronal Autophagy and Apoptosis via the PI3K/AKT Pathway Following Ischemic Stroke.
Electroacupuncture (EA) is a safe and effective therapy for ischemic stroke in both clinical and laboratory settings. However, the underlying mechanism behind EA treatment for stroke remains unclear. Here, we aimed to evaluate whether EA treatment at the acupoints of Zusanli (ST36) and Quchi (LI11) exerted a neuroprotective effect on ischemic stroke rats by modulating autophagy and apoptosis via the PI3K/AKT/mTOR signaling pathway. EA was performed at 24 h following brain ischemia/reperfusion (I/R) for 30 min per day for 3 days. Our results indicated that EA treatment significantly decreased neurological deficits and cerebral infarct volume in ischemic stroke rats. Also, EA intervention markedly reduced neuronal apoptosis by suppressing the activation of cleaved caspase-3 (CCAS3) at 72 h following I/R, as shown by a Western blot analysis. Furthermore, EA treatment after ischemic stroke suppressed the ischemia activated expression level of LC3II/I and Atg7 and increased the ischemia inhibited expression level of PI3K, phosphorylation of mTOR, phosphorylation of AKT, P62 and LAMP1, hence mediating the autophagy level of the neurocyte, which was reversed by the PI3K inhibitor Dactolisib. In summary, our results indicate that the protective effects of EA treatment at points of Quchi (LI11) and Zusanli (ST36) in rats following cerebral I/R injury was associated with the inhibition of neuronal apoptosis and autophagy via activating the PI3K/AKT/mTOR signaling pathway.