Involvement of 5-hydroxytryptamine-containing neurons in antinociception produced by injection of morphine into nucleus raphe magnus or onto spinal cord.
We studied whether antinociception produced by injection of morphine into the nucleus raphe magnus (NRM) or superfusion onto the spinal cord involved serotonergic neurons that descend from brainstem to spinal cord. Involvement of 5-hydroxytryptamine (5-HT)-containing neurons was determined by correlating morphine-induced analgesia with an increase in turnover of 5-HT and by determining if depletion of cord 5-HT with the neurotoxin, 5,7-dihydroxytryptamine (5,7-DHT) could attenuate the antinociceptive effects of morphine. When injected directly into the NRM, 10 micrograms of morphine produced profound analgesia as measured by the paw-pressure technique, and significantly increased the turnover of 5-HT in both posterior medulla and spinal cord. Depletion of cord 5-HT to less than 10% of control concentrations attenuated the antinociceptive effect of morphine injected into the NRM. When various concentrations of morphine (1, 10 or 50 micrograms) were injected directly into the spinal subarachnoid space, a dose-dependent analgesia was observed. No change in 5-HT turnover in spinal cord was observed with any dose of morphine superfused onto the cord. In addition, depletion of cord 5-HT with 5,7-DHT did not alter the analgesic response to either 1 or 10 micrograms of intrathecal morphine. These results suggest that although 5-HT-containing neurons descending from brainstem into spinal cord are involved with analgesia produced by morphine injection into the NRM, they are not involved in the analgesia induced by applying morphine directly to the cord.


Chemical constituents isolated from Polygala japonica leaves and their inhibitory effect on nitric oxide production in vitro.
A methanolic extract of dried leaves of Polygala japonica Houtt (Polygalaceae) significantly attenuated nitric oxide production in lipopolysaccharide-simulated BV2 microglia. Five anthraquinones chrysophanol (1), emodin (2), aloe-emodin (3), emodin 8-O-beta-D-glucopyranoside (4) and trihydroxy anthraquinone (5), and four flavonoids kaempferol (6), chrysoeriol (7), kaempferol 3-gentiobioside (8) and isorhamnetin (9) were isolated from the methanolic extract using bioactivity-guided fractionation. Among them, compounds 1-4, 6 and 7 showed significant inhibitory effect on lipopolysaccharide-induced nitric oxide production in BV2 microglia at the concentrations ranging from 1.0 to 100.0 microM.


The first international standard for human urinary FSH and for human urinary LH (ICSH), for bioassay.
This paper describes the preparation and nature of the First International Standard for Human Urinary Follicle Stimulating Hormone and for Human Urinary Luteinizing Hormone, for Bioassay, and of two batches of working standard which were prepared from the same material. A collaborative study of these materials was carried out by six laboratories in six different countries. The FSH and LH activities of the Standard were assayed in terms of those of the Second International Reference Preparation of Human Menopausal Gonadotrophins, Urinary, for Bioassay, which it replaces. The results from 20 valid FSH assays and 30 valid LH assays (using four different methods) obtained in this way gave a weighted combined potency estimate for FSH of 53.7 IU, with 95 % fiducial limits of 47.2-61.1 IU, and for LH of 46.2 IU, with 95 % fiducial limits of 43.3-49.3 IU. Accelerated degradation studies of the Standard stored at elevated temperatures suggested that the stability of both FSH and LH activities under normal storage conditions would be satisfactory. The FSH and LH activities of the two batches of working standard (WS-A and WS-B) were compared with those of the Standard and were not found to differ significantly, except for the LH activity of WS-B which appeared to be slightly higher than that of the Standard. Accelerated degradation studies did not show any significant differences in stability between the Standard and batches of working standards. On the basis of these results the Standard has been established by WHO and allocated a potency for FSH of 54 IU per ampoule and for LH 46 IU per ampoule. The International Units for FSH, Human Urinary, for Bioassay and for LH (ICSH), Human Urinary, for Bioassay are thus defined as the activities contained in 0.1138 mg and 0.13369 mg of the International Standard, respectively.


Properties of a class C beta-lactamase from Serratia marcescens.
A beta-lactamase produced by a penicillin-resistant strain of Serratia marcescens was isolated and purified. The kcat. value for benzylpenicillin was about 5% of that observed for the best cephalosporin substrates. However, the low Km of the penam resulted in a high catalytic efficiency (kcat./Km) and the classification of the enzyme as a cephalosporinase might not be completely justified. It also exhibited a low but measurable activity against cefotaxime, cefuroxime, cefoxitin and moxalactam. Substrate-induced inactivation was observed both with a very good (cephalothin) or a very bad (moxalactam) substrate. The active site was labelled by beta-iodopenicillanate. Trypsin digestion produced a 19-residue active-site peptide whose sequence clearly allowed the classification of the enzyme as a class C beta-lactamase.


Fast variational alignment of non-flat 1D displacements for applications in neuroimaging.
In the context of signal analysis and pattern matching, alignment of 1D signals for the comparison of signal morphologies is an important problem. For image processing and computer vision, 2D optical flow (OF) methods find wide application for motion analysis and image registration and variational OF methods have been continuously improved over the past decades. We propose a variational method for the alignment and displacement estimation of 1D signals. We pose the estimation of non-flat displacements as an optimization problem with a similarity and smoothness term similar to variational OF estimation. To this end, we can make use of efficient optimization strategies that allow real-time applications on consumer grade hardware. We apply our method to two applications from functional neuroimaging: The alignment of 2-photon imaging line scan recordings and the denoising of evoked and event-related potentials in single trial matrices. We can report state of the art results in terms of alignment quality and computing speeds. Existing methods for 1D alignment target mostly constant displacements, do not allow native subsample precision or precise control over regularization or are slower than the proposed method. Our method is implemented as a MATLAB toolbox and is online available. It is suitable for 1D alignment problems, where high accuracy and high speed is needed and non-constant displacements occur.


The relative importance of salinization in lowland stream zooplankton: Implications of the ecosystem nutrient status.
Salinization of aquatic systems is predicted to increase due to climate and land use changes. Nevertheless, community responses may be different according to the ecosystem characteristics and contextual scenarios. Small flowing waters are particularly vulnerable to salinization, which may impact on the biodiversity and ecosystem processes, but this remains unclear. We conducted a study in 42 lowland streams characterized by overall high nutrient levels along a salinity gradient between 2 and 160 g L-1 to analyze changes in zooplankton structural and functional metrics, and the grazing effects of zooplankton on phytoplankton affecting the energy transfer. Generalized additive models revealed that the analyzed metrics were relatively influenced by salinity, with factors related to trophic conditions playing an important role as well. Total abundance and biomass decreased along the salinity gradient while increasing at intermediate soluble reactive phosphorous concentrations (SRP) in the former and with a linear increase in the SRP in the latter case. Taxonomic richness decreased with salinity and dissolved inorganic nitrogen, with species replacement toward saline-tolerant ones according to the compositional and optimums analyses. In opposite, functional richness did not display any specific trend within the environmental gradients. This explains why zooplankton compositional changes were not reflected into shifts in the grazing pattern on phytoplankton, which was in turn driven by SRP and dissolved oxygen concentrations. Further research is a critical requirement in these poorly studied ecosystems for planning mitigation actions to the co-occurrence of eutrophication and salinization in a fast changing world.


Real-world trends in pre-dialysis blood pressure levels of patients undergoing dialysis in Japan using a web-based national database.
The prevalence of hypertension is high among patients undergoing dialysis. We extracted data of patients undergoing dialysis between 2012 and 2020 with recorded pre-dialysis systolic blood pressure (SBP) using a web-based national database in Japan. Following the 2019 Japanese Society of Hypertension guidelines, we classified SBP and assessed its trends over time based on sex, age, diabetes status, and the anti-hypertensive medication use. Using the 2020 database, we examined 336,759 Japanese patients undergoing dialysis (114,249 female; 222,510 male). The mean age was 69.4 ± 12.5 years, and the mean SBP was 152.3 ± 24.7 mm Hg. The prevalence rate of pre-dialysis hypertension was 70.2%, with 32.5%, 24.5%, and 13.2% of patients having grade I, grade II, and grade III hypertension, respectively. From 2014 to 2020, prevalence rate of pre-dialysis hypertension and absolute values of pre-dialysis SBP were higher in dialysis patients with diabetes than in those without diabetes across all age groups and sexes. Younger patients with diabetes or those on anti-hypertensive medication exhibited an SBP of approximately 160 mm Hg. Cerebrovascular death in patients with diabetes was associated with a higher rate of pre-dialysis hypertension than that in those without diabetes, and there was a significant difference in the prevalence of grade III hypertension between the two groups. In conclusion, the mean pre-dialysis SBP among patients undergoing dialysis remained high, and younger patients with diabetes or those receiving anti-hypertensive medications had poor blood pressure control. Optimal blood pressure management may be necessary to reduce the risk of cardiovascular mortality.


Theoretical studies on CO2 capture behavior of quaternary ammonium-based polymeric ionic liquids.
Quaternary ammonium-based polymeric ionic liquids (PILs) are novel CO2 sorbents as they have high capacity, high stability and high binding energy. Moreover, the binding energy of ionic pairs to CO2 is tunable by changing the hydration state so that the sorbent can be regenerated through humidity adjustment. In this study, theoretical calculations were conducted to reveal the mechanism of the humidity swing CO2 adsorption, based on model compounds of quaternary ammonium cation and carbonate anions. The electrostatic potential map demonstrates the anion, rather than the cation, is chemically preferential for CO2 adsorption. Further, the proton transfer process from water to carbonate at the sorbent interface is successfully depicted with an intermediate which has a higher energy state. By determining the CO2 adsorption energy and activation energy at different hydration states, it is discovered that water could promote CO2 adsorption by reducing the energy barrier of proton transfer. The adsorption/desorption equilibrium would shift to desorption by adding water, which constitutes the theoretical basis for humidity swing. By analyzing the hydrogen bonding and structure of the water molecules, it is interesting to find that the CO2 adsorption weakens the hydrophilicity of the sorbent and results in release of water. The requirement of latent heat for the phase change of water could significantly reduce the heat of adsorption. The special "self-cooling" effect during gas adsorption can lower the temperature of the sorbent and benefit the adsorption isotherms.


Predicting bulk mechanical properties of cellularized collagen gels using multiphoton microscopy.
Cellularized collagen gels are a common model in tissue engineering, but the relationship between the microstructure and bulk mechanical properties is only partially understood. Multiphoton microscopy (MPM) is an ideal non-invasive tool for examining collagen microstructure, cellularity and crosslink content in these gels. In order to identify robust image parameters that characterize microstructural determinants of the bulk elastic modulus, we performed serial MPM and mechanical tests on acellular and cellularized (normal human lung fibroblasts) collagen hydrogels, before and after glutaraldehyde crosslinking. Following gel contraction over 16 days, cellularized collagen gel content approached that of native connective tissues (∼200 mg ml⁻¹). Young's modulus (E) measurements from acellular collagen gels (range 0.5-12 kPa) exhibited a power-law concentration dependence (range 3-9 mg ml⁻¹) with exponents from 2.1 to 2.2, similar to other semiflexible biopolymer networks such as fibrin and actin. In contrast, cellularized collagen gel stiffness (range 0.5-27 kPa) produced concentration-dependent exponents of 0.7 uncrosslinked and 1.1 crosslinked (range ∼5-200 mg ml⁻¹). The variation in E of cellularized collagen hydrogels can be explained by a power-law dependence on robust image parameters: either the second harmonic generation (SHG) and two-photon fluorescence (TPF) (matrix component) skewness (R²=0.75, exponents of -1.0 and -0.6, respectively); or alternatively the SHG and TPF (matrix component) speckle contrast (R²=0.83, exponents of -0.7 and -1.8, respectively). Image parameters based on the cellular component of TPF signal did not improve the fits. The concentration dependence of E suggests enhanced stress relaxation in cellularized vs. acellular gels. SHG and TPF image skewness and speckle contrast from cellularized collagen gels can predict E by capturing mechanically relevant information on collagen fiber, cell and crosslink density.


Streptococcus mutans promotes tumor progression in oral squamous cell carcinoma.
Oral squamous cell carcinoma (OSCC) is an aggressive head and neck cancer. Evidence showed that some pathogenic bacteria are associated with periodontitis and oral cancer. The change in oral microbiome composition and the role of the specific periodontal pathogen Streptococcus mutans in OSCC were investigated. We analyzed the microbiome of oral biofilms to identify if the oral microbiome composition was associated with OSCC. The role of S. mutans with clinical prognosis for OSCC was also examined. We further examined the role of S. mutans infection in OSCC progression in preclinical experiments. The microbiome assay by oral biofilms revealed that there was different microbiota composition between OSCC patients and health participants. Furthermore, the microbiota profiles showed that S. mutans abundance was associated with the development of OSCC development. Using the 16S rRNA PCR analysis, the presence of S. mutans was associated with advanced clinical stage and poor disease control. Furthermore, in the 4-nitroquinoline 1-oxide-induced mouse model, the presence of S. mutans was associated with elevated invasive oral cancer incidence. By cellular and xenograft tumor model using oral cancer cells, S. mutans infection was associated with the increased tumor aggressiveness, the epithelial-mesenchymal transition and interleukin-6 (IL-6) production; it also correlated with the recruitment of myeloid-derived-suppressor cells. When IL-6 signaling inhibited, the effects of S. mutans on tumor aggressiveness were attenuated. In conclusion, S. mutans may have the additive effect on oral cancer development and progression. Good oral hygiene to eradicate S. mutans or targeting IL-6 signaling could be a promising strategy for OSCC associated with S.mutans infection.


A review and phylogeny of Scarabaeine dung beetle fossils (Coleoptera: Scarabaeidae: Scarabaeinae), with the description of two Canthochilum species from Dominican amber.
Despite the increasing rate of systematic research on scarabaeine dung beetles (Coleoptera: Scarabaeidae: Scarabaeinae), their fossil record has remained largely unrevised. In this paper, we review all 33 named scarabaeine fossils and describe two new species from Dominican amber (Canthochilum alleni sp.n., Canthochilum philipsivieorum sp.n.). We provide a catalogue of all fossil Scarabaeinae and evaluate their assignment to this subfamily, based primarily on the original descriptions but also, where possible, by examining the type specimens. We suggest that only 21 fossil taxa can be reliably assigned to the Scarabaeinae, while the remaining 14 should be treated as doubtful Scarabaeinae. The doubtful scarabaeines include the two oldest dung beetle fossils known from the Cretaceous and we suggest excluding them from any assessments of the minimum age of scarabaeine dung beetles. The earliest reliably described scarabaeine fossil appears to be Lobateuchus parisii, known from Oise amber (France), which shifts the minimum age of the Scarabaeinae to the Eocene (53 Ma). We scored the best-preserved fossils, namely Lobateuchus and the two Canthochilum species described herein, into the character matrix used in a recent morphology-based study of dung beetles, and then inferred their phylogenetic relationships with Bayesian and parsimony methods. All analyses yielded consistent phylogenies where the two fossil Canthochilum are placed in a clade with the extant species of Canthochilum, and Lobateuchus is recovered in a clade with the extant genera Ateuchus and Aphengium. Additionally, we evaluated the distribution of dung beetle fossils in the light of current global dung beetle phylogenetic hypotheses, geological time and biogeography. The presence of only extant genera in the late Oligocene and all later records suggests that the main present-day dung beetle lineages had already been established by the late Oligocene-mid Miocene.


An integrated model of care for inflammatory bowel disease sufferers in Australia: development and the effects of its implementation.
Psychological comorbidities are associated with poor outcome and increased healthcare utilization in patients with inflammatory bowel disease (IBD). However, a model of care addressing the biopsychosocial dimension of disease is not routinely applied in IBD. This review describes the development of such a model and the effects of its implementation in a hospital-based cohort of patients with IBD. Three different approaches were used: 1) collecting baseline epidemiological data on mental health comorbidities; 2) raising awareness of and targeting mental health problems; 3) examining the effects of the model implementation. High rates of anxiety and depressive symptoms (36% and 13%, respectively) that are maintained over time were identified in IBD patients presenting at a metropolitan teaching hospital. Patients with documented psychological comorbidities were more likely to be hospitalized than those without (odds ratio [OR] = 4.13, 95% confidence interval [CI]: 1.25, 13.61). Improvements in disease activity, anxiety, depression, quality of life, and coping have been noted when cognitive-behavioral therapy (CBT) was provided to patients. A drop in the use of opiates (P = 0.037) and hospitalization rates (from 48% to 30%) in IBD patients has been noted as a result of introduction of the changed model of care. In addition, the mean total cost of inpatient care was lower for IBD patients than controls (US$12,857.48 [US$15,236.79] vs. US$ 30,467.78 [US$ 53,760.20], P = 0.005). Our data to date suggest that an integrated model of care for patients with IBD may yield superior long-term outcomes in terms of medication use and hospitalization rates and reduce healthcare costs.


What Potentially Modifiable Factors are Associated With Treatment Nonadherence in Pediatric Growth Hormone Deficiency? A Quantitative Study.
A recent systematic review reported that up to 71% of patients with growth hormone deficiency and their families are nonadherent to treatment as prescribed. Nonadherence to growth hormone treatment presents a substantial and costly problem for the patient, health care provider, and health care system. The current study uniquely investigated the potentially modifiable factors associated with treatment nonadherence in this endocrine disorder. The cross-sectional study was conducted among 82 parent/caregivers of children with growth hormone deficiency who were receiving growth hormone treatment. Self-report questionnaires investigated parent/caregiver perceptions and experiences of their child's condition and prescribed treatment, in addition to their perceived relationship with their health care professional. The 8-item Morisky medication adherence scale was used for the assessment of treatment adherence. Sixty-two percent of parents/caregivers were found to be nonadherent to growth hormone treatment as prescribed. Illness perceptions (consequences, identity, and coherence) and treatment concerns were found to be significantly associated with treatment adherence, as was the quality of the health care professional-parent/caregiver relationship. The study confirmed the extent of the adherence problem evident among the pediatric growth hormone deficiency population. In addition, it presented an insight into the explanatory factors that underpin nonadherence to growth hormone treatment. Our findings can be used to inform the development of adherence-focused interventions, with the purpose of supporting patients and their families and improving the use of prescribed growth hormone treatment within endocrine clinical practice.


Competing regimes of motion of 1D mobile impurities.
We show that a distinguishable mobile impurity inside a one-dimensional many-body state at zero temperature generally does not behave like a quasiparticle. Instead, both the impurity dynamics as well as the ground state of the bath are fundamentally transformed by a diverging number of zero-energy excitations being generated, leading to what we call infrared-dominated (ID) dynamics. Combining analytics and density matrix renormalization group numerics, we provide a general formula for the power law governing ID dynamics at zero momentum, discuss a threshold beyond which quasiparticle dynamics may occur again, and study the competition between the ID and quasiparticle universality classes at larger impurity momenta.


Vitamin D receptor gene polymorphism predicts height and bone size, rather than bone density in children and young adults.
Peak bone mass is considered to be under strong genetic control. We studied the association among anthropometry, bone density and vitamin D receptor (VDR) genotype in an ethnically homogeneous group of 148 Caucasian children and young adults. Bone density was measured by dual energy X-ray absorptiometry (DXA) and VDR genotype was determined by a direct haplotyping procedure of the BsmI, ApaI, and TaqI restriction fragment length polymorphisms. A second DXA measurement was made after approximately 4 years. Results are expressed as age- and sex-adjusted standard deviation scores (SDS). Previously, the collagen IA1 Sp1 polymorphism was studied in this population. We found VDR genotype to be associated with a 0.4 SDS increased height per allele copy of haplotype '3' (P = 0.04) and a 0.4 SDS increased width of the lumbar vertebral body in the haplotype '3' allele carriers (P = 0.05). We observed a trend towards a 0.3 SDS decreased bone mineral apparent density of lumbar spine (BMAD) per copy of haplotype '3' allele (P = 0.10). In contrast, no association with areal bone mineral density (BMD) was observed. In the follow-up analyses, no differences in height or bone gain among the VDR genotypes were demonstrated. By combining the risk alleles of VDR and collagen IA1 Sp1 genotype, an additive genotype effect on height (P = 0.006) and vertebral body width (P = 0.001) was found. In this exploratory study we found VDR genotype to be associated with frame size and BMAD. The VDR genotype effects on stature and bone size seem to neutralize the effect on areal BMD.


A two-stage subsurface vertical flow constructed wetland for high-rate nitrogen removal.
By using a two-stage constructed wetland (CW) system operated with an organic load of 40 gCOD.m(-2).d(-1) (2 m2 per person equivalent) average nitrogen removal efficiencies of about 50% and average nitrogen elimination rates of 980 g N.m(-2).yr(-1) could be achieved. Two vertical flow beds with intermittent loading have been operated in series. The first stage uses sand with a grain size of 2-3.2 mm for the main layer and has a drainage layer that is impounded; the second stage sand with a grain size of 0.06-4 mm and a drainage layer with free drainage. The high nitrogen removal can be achieved without recirculation thus it is possible to operate the two-stage CW system without energy input. The paper shows performance data for the two-stage CW system regarding removal of organic matter and nitrogen for the two year operating period of the system. Additionally, its efficiency is compared with the efficiency of a single-stage vertical flow CW system designed and operated according to the Austrian design standards with 4 m2 per person equivalent. The comparison shows that a higher effluent quality could be reached with the two-stage system although the two-stage CW system is operated with the double organic load or half the specific surface area requirement, respectively. Another advantage is that the specific investment costs of the two-stage CW system amount to 1,200 EUR per person (without mechanical pre-treatment) and are only about 60% of the specific investment costs of the singe-stage CW system.


Neural Stem/Progenitor Cell (NSPC) Extraction and Culture.
Neural stem-progenitor cells (NSPCs) are multipotent, self-renewing cells that generate radial glial cells (RGC). RGCs then give rise to neurons and glia during neural development. Here, we describe the process of NSPC isolation and culturing to form clonal aggregates termed neurospheres. There are multiple assays outlined in this chapter that allow us to quantify differences in proliferation, self-renewal potential, and differentiation of these cells.


Glucose counterregulatory hormones in the 72-hour fast.
To attempt to establish a reference range of glucagon concentrations during hypoglycemia. We measured glucagon, cortisol, and growth hormone responses in 65 patients with insulinoma and 29 normal control subjects who underwent a 72-hour fast. For comparison, we also assessed these responses in eight patients with noninsulinoma pancreatogenous hypoglycemia syndrome. At the end of the fasts, median serum cortisol (19.0 mg/dL [range, 3.7 to 44.0] versus 11.0 mg/dL [range, 5.0 to 28.0], respectively; P<0.001) and growth hormone levels (3.5 ng/mL [range, 0.1 to 46.0] versus 1.2 ng/mL [range, 0.1 to 34.0], respectively; P = 0.021) were higher, whereas plasma glucagon (54.5 pg/mL [range, 11.0 to 170.0] versus 75.0 pg/mL [range, 17.0 to 940.0], respectively; P = 0.012) was lower in patients with insulinoma (serum glucose level, 39 mg/dL [range, 14 to 58]) than in control subjects (serum glucose level, 63 mg/dL [range, 47 to 89]). In contrast, the 8 patients with noninsulinoma pancreatogenous hypoglycemia syndrome, a disorder of postprandial hyperinsulinemic hypoglycemia with normal findings on a 72-hour fast (serum glucose level, 71.5 mg/dL [range, 48 to 82]), had concentrations of glucagon (81.0 pg/mL [range, 47.0 to 150.0]), cortisol (10.5 mg/dL [range, 2.7 to 17.0]), and growth hormone (1.5 ng/mL [range, 0.8 to 6.9]) similar to those in the control subjects. On multivariate analysis, the duration of the fast, baseline glucagon concentration, and male gender (but not age, body mass index, or concentrations of glucose and insulin) were correlated with end-of-fast glucagon concentration. Defective glucagon secretion in hypoglycemic disorders applies to the stimulus of hypoglycemia but not to food deprivation. A conservative estimate for glucagon deficiency based on the minimal observed glucagon response could be a level of 10 pg/mL during hypoglycemia.


Temporal trends in prevalence of bacteria isolated from foals with sepsis: 1979-2010.
Sepsis is an important cause of death in foals. Knowledge of which pathogens are likely to be involved is important for selection of antimicrobial drugs for initial treatment. To identify temporal trends in prevalence of bacteria isolated from foals with sepsis between 1979 and 2010. Retrospective review of medical records. All foals ≤30 days of age presented to the Veterinary Medical Teaching Hospital (VMTH) at the University of California, Davis between 1979 and 2010, with a diagnosis of sepsis confirmed by culture of bacteria from blood or internal organs (antemortem or at necropsy), were included in the study. Conventional microbiological methods were used to identify isolated organisms. The Cochran-Armitage trend test was used for statistical analysis. The percentage of Gram-positive isolates increased significantly over the years. The percentage Enterobacteriacea, and Klebsiella spp. in particular, decreased over time. Enterococcus spp. isolates were cultured more often in recent years. Whereas Gram-negative bacteria, particularly Enterobacteriaceae, remain the most common isolates from neonatal foals with sepsis, the prevalence of Gram-positive bacteria is increasing. This trend underlines the importance of including antimicrobial drugs active against both Gram-positive and Gram-negative bacteria in treatment protocols while awaiting the results of bacteriological culture and susceptibility tests. The increased prevalence of Enterococcus spp. is of concern because antimicrobial susceptibility patterns for enterococci are unpredictable and enterococci can also act as donors of antimicrobial resistance genes to other bacteria.


Flow cytometry based enumeration and functional characterization of CD8 T regulatory cells in patients with multiple myeloma before and after lenalidomide plus dexamethasone treatment.
Multiple myeloma (MM) is a malignancy of plasma cells frequently associated with immune abnormalities. Several studies have confirmed that in MM immune deregulation can be mediated by increased numbers of CD4 T regulatory (Treg) cells, and these cells were also associated with poor outcome. In the present study, we aimed to study CD8 Treg cells before and after lenalidomide plus dexamethasone (len-dex) treatment in MM patients. Using flow cytometry, we enumerated and assessed suppressive function of CD8 Treg cells in 16 MM patients before and after len-dex treatment. Numbers of CD8 Treg cells (CD8+CD25hi+FoxP3+) (P<0.01) were significantly increased in MM patients (before treatment) compared to healthy donors. However, no significant changes were observed in CD4 and CD8 T cells. A significant increase in CD8 Treg cells was observed after len-dex treatment compared to pre-treatment but no significant difference was observed in CD4 and CD8 T cells. Proliferation assay data showed that CD8 Treg cells inhibited proliferation of CD4 T cells and IFN-γ secretion in a concentration dependent manner. Suppressive activity of CD8 Treg cells did not differ significantly between healthy donors, untreated and len-dex treated MM patients. A significant abnormal level of IL-10 was observed from proliferation assays of untreated and len-dex treated MM patients compared to healthy donors (P≤0.03). Using flow cytometry, we have shown that suppressive CD8 Treg cells are increased in MM patients and len-dex treatment is unable to control these suppressive CD8 Treg cells. © 2013 Clinical Cytometry Society.


Voltage-gated sodium channels: biophysics, pharmacology, and related channelopathies.
Voltage-gated sodium channels (VGSC) are multi-molecular protein complexes expressed in both excitable and non-excitable cells. They are primarily formed by a pore-forming multi-spanning integral membrane glycoprotein (α-subunit) that can be associated with one or more regulatory β-subunits. The latter are single-span integral membrane proteins that modulate the sodium current (I(Na)) and can also function as cell adhesion molecules. In vitro some of the cell-adhesive functions of the β-subunits may play important physiological roles independently of the α-subunits. Other endogenous regulatory proteins named "channel partners" or "channel interacting proteins" (ChiPs) like caveolin-3 and calmodulin/calmodulin kinase II (CaMKII) can also interact and modulate the expression and/or function of VGSC. In addition to their physiological roles in cell excitability and cell adhesion, VGSC are the site of action of toxins (like tetrodotoxin and saxitoxin), and pharmacologic agents (like antiarrhythmic drugs, local anesthetics, antiepileptic drugs, and newly developed analgesics). Mutations in genes that encode α- and/or β-subunits as well as the ChiPs can affect the structure and biophysical properties of VGSC, leading to the development of diseases termed sodium "channelopathies". This review will outline the structure, function, and biophysical properties of VGSC as well as their pharmacology and associated channelopathies and highlight some of the recent advances in this field.


The Relationship Between Lesion Localization and Dysphagia in Acute Stroke.
Factors that can facilitate early identification of individuals at risk of dysphagia such as stroke location are potentially of great benefit. The aim of this study was to examine the role of hemisphere and lesion location in assessing dysphagia pattern and airway invasion as identified through the use of validated, standardized interpretation measures for the videofluoroscopic swallowing study. Consecutive patients (N = 80) presenting with stroke symptoms who had a first-time acute ischemic stroke confirmed on diffusion-weighted magnetic resonance imaging (DW-MRI) scan participated. Three swallowing outcome variables were assessed using regression models: modified barium swallow impairment profile (MBSImP) oral impairment (OI) and pharyngeal impairment (PI) scores and penetration aspiration scale (PAS) score. Subjects were primarily male and demonstrated mild stroke and mild to moderate dysphagia. There was a significantly higher likelihood of abnormal PAS scores for infratentorial lesions compared to right hemisphere location (Odds ratio: 3.1, SE: 1.8, p = 0.046) and for Whites compared to African Americans (Odds ratio: 5.5, SE: 2.6, p = <0.001). However, OI scores were higher (worse) in African Americans compared to Whites (Beta = -1.2; SE: 0.56; p = 0.037). PI scores had no significant association with race or lesion location. Using DW-MRI to identify infratentorial stroke can help identify individuals at risk of airway invasion; however, imaging information concerning supratentorial infarct hemisphere and location may not be useful to predict which individuals with mild stroke are at risk for dysphagia and aspiration when admitted with acute stroke symptoms. Future studies should explore the role of race in the development of stroke-related dysphagia.


PhiC31 integrase-mediated cassette exchange in silkworm embryos.
To construct an effective site-specific integration system in the silkworm, we examined if phiC31 integrase works in silkworm embryos. As an assay system, we constructed an extrachromosomal cassette exchange reaction system between two attP sites of an acceptor plasmid and two attB sites of a donor plasmid. To evaluate the activity, integrase mRNAs synthesized from three different plasmids were used. We injected a mixture of the acceptor and donor plasmids with the mRNA synthesized in vitro from one of the three plasmids into silkworm embryos at 4-6 h after oviposition and recovered plasmid DNAs from the embryos 3 days after injection. The resultant plasmids were transformed into Escherichia coli and spread on selection medium plates containing the appropriate antibiotics. A colony-forming assay and restriction enzyme digestion of the plasmids purified from the colonies showed that the phiC31 integrase worked very efficiently in the silkworm embryos. Notably, a phiC31 integrase mRNA synthesized from two of the plasmids produced cassette exchange plasmids at a high frequency, suggesting that the mRNA can be used to construct a targeted integration system in silkworms.


Lack of survival benefit following adjuvant radiation in patients with retroperitoneal sarcoma: a SEER analysis.
The benefit of radiation therapy (RT) among patients with retroperitoneal sarcoma (RPS) is controversial. We performed a retrospective analysis of the effect of RT on survival among RPS patients using a nationwide cancer registry. Utilizing data from the Surveillance, Epidemiology, and End Results (SEER) database, we identified 2308 cases of RPS from 1988 to 2004. We excluded 773 cases for age < 18, identification by autopsy only, absence of histologic confirmation, presence of metastatic disease, or lack of surgical intervention. Overall survival (OS) and disease-specific survival (DSS) were estimated using the Kaplan-Meier method. Multivariate analysis was performed using a Cox proportional hazards model, adjusting for significant covariables. Among 1535 patients who met entry criteria, RT was administered to 373 patients (24.3%). The majority of RT (n = 300, 80.4%) was administered postoperatively. Median OS was 60 and 60 mo, respectively, for patients receiving and not receiving RT (P = 0.59). Median DSS was 86 and 117 mo, respectively, for patients receiving and not receiving RT (P = 0.84). On multivariate analysis, younger age, female gender, low and intermediate histologic grade, liposarcoma histology, tumor size 5-10 cm, and completeness of resection all independently predicted better OS and DSS, while RT did not (HR for OS with RT 0.92, 95% CI 0.78-1.09 and HR for DSS with RT 0.96, 95% CI 0.78-1.17). On subgroup analysis by histology, patients with malignant fibrous histiocytoma (MFH) receiving RT demonstrated statistically improved OS (P = 0.002) and DSS (P = 0.01), respectively. With the possible exception of MFH, postoperative RT offers no survival benefit in RPS. Further studies are necessary to determine if the selective application of RT is indicated.


[Craniocerebral trauma and skiing: the ABC of the general surgeon].
This paper describes the classical encephalic lesions due to ski accidents, after a short review of the normal cerebral physiology. It analyses the main steps of the management of the head injury patient by the general surgeon taking history, general and neurologic state, diagnosis, first aid, initial medication, transport conditions and information of the neurosurgical center.


Machine learning as the new approach in understanding biomarkers of suicidal behavior.
In psychiatry, compared to other medical fields, the identification of biological markers that would complement current clinical interview, and enable more objective and faster clinical diagnosis, implement accurate monitoring of treatment response and remission, is grave. Current technological development enables analyses of various biological marks in high throughput scale at reasonable costs, and therefore 'omic' studies are entering the psychiatry research. However, big data demands a whole new plethora of skills in data processing, before clinically useful information can be extracted. So far the classical approach to data analysis did not really contribute to identification of biomarkers in psychiatry, but the extensive amounts of data might get to a higher level, if artificial intelligence in the shape of machine learning algorithms would be applied. Not many studies on machine learning in psychiatry have been published, but we can already see from that handful of studies that the potential to build a screening portfolio of biomarkers for different psychopathologies, including suicide, exists.


Safe Return to Exercise after COVID-19 Infection.
COVID-19 has been posing a significant challenge to the athletic community. A wide spectrum of clinical features was reported in athletes with confirmed COVID-19 infection having mild to severe symptoms requiring hospitalisation and intensive medical care. Possibly, a lack of exercise may negatively impact the immune system, leading to an increased susceptibility to infection and comorbidities. Determining safe return to exercise after COVID-19 infection is a challenge for sports physicians. A gradual return to exercise under the guidance of a specialised medical team to achieve the pre-infection fitness level is of utmost importance. Individualised exercise programmes must be initiated at low intensity which could be gradually made more rigorous based on the pertinent metabolic equivalent. Thus, there is a clear need for clinical recommendations before athletes may return to exercise after COVID-19 infection.


Strengthening and sustainability of national immunization technical advisory groups (NITAGs) globally: Lessons and recommendations from the founding meeting of the global NITAG network.
National Immunization Technical Advisory Groups (NITAGs) provide independent, evidence-informed advice to assist their governments in immunization policy formation. However, many NITAGs face challenges in fulfilling their roles. Hence the many requests for formation of a network linking NITAGs together so they can learn from each other. To address this request, the Health Policy and Institutional Development (HPID) Center (a WHO Collaborating Center at the Agence de Médecine Préventive - AMP), in collaboration with WHO, organized a meeting in Veyrier-du-Lac, France, on 11 and 12 May 2016, to establish a Global NITAG Network (GNN). The meeting focused on two areas: the requirements for (a) the establishment of a global NITAG collaborative network; and (b) the global assessment/evaluation of the performance of NITAGs. 35 participants from 26 countries reviewed the proposed GNN framework documents and NITAG performance evaluation. Participants recommended that a GNN should be established, agreed on its governance, function, scope and a proposed work plan as well as setting a framework for NITAG evaluation.


Functional prostaglandin-endoperoxide synthase 2 polymorphism predicts poor outcome in sarcoidosis.
The majority of patients with sarcoidosis resolve their condition; however 5-10% of patients with sarcoidosis develop pulmonary fibrosis with poor prognosis. Prostaglandin-endoperoxide synthase 2 (PTGS2) is a key regulatory enzyme in the synthesis of the antifibrotic agent prostaglandin E(2) and is reduced in sarcoidosis lung. A promoter polymorphism in PTGS2, -765G>C, is reported to reduce its expression. To investigate if -765G>C is associated with susceptibility to, and poorer outcome within, sarcoidosis and to examine a possible mechanism by which -765G>C reduces PTGS2 expression. We used a case-control design study and genotyped -765G>C in a white British population of 198 patients with sarcoidosis and 166 control subjects. Patients with sarcoidosis were classified before genotyping as having persistent or nonpersistent disease using clinical criteria that included chest radiography staging, need for treatment, lung function, and longitudinal follow-up. Electrophoretic mobility shift assays were used to identify changes in transcription factor binding caused by the -765G>C polymorphism. Carriage of the -765C allele was strongly associated with susceptibility to sarcoidosis (odds ratio, 2.50; 95% confidence interval, 1.51-4.13; p=0.006) and, within this disease, with poorer outcome (odds ratio, 3.11; 95% confidence interval, 1.35-7.13; p=0.008). The association with sarcoidosis was replicated in a second Austrian population. Electrophoretic mobility shift assays revealed that the -765C allele causes a loss of Sp1/Sp3 transcription factor binding and an increase in Egr-1 binding to the region. Our data suggest that the -765G>C polymorphism identifies individuals who are susceptible to sarcoidosis and, more importantly, at risk of pulmonary fibrotic disease. An altered Sp1/Sp3 binding to the -765 region may contribute to the mechanism by which -765G>C reduces PTGS2 expression.


[Genotoxic effects of tonarol].
Genotoxic effects of 2,6-di-tret-butyl-4-methylphenol (tonarol) were studied using four test systems: the Ames test, the SOS chromotest, the cytogenetic test with rootlets of onion (Allium cepa), and the in vivo micronucleus test. Tonarol did not affect gene mutation induction in Salmonella typhimurium tester strains, the SOS response in the Escherichia coli strain PQ37, chromosomal aberrations in cells of onion (Allium cepa) rootlets, and micronuclei in erythrocytes of peripheral blood of CBA x C5713 L/G mice. Tonarol induced cell division in A.


Early Fasciotomy and Limb Salvage and Complications in Military Lower Extremity Vascular Injury.
Military guidelines endorse early fasciotomy after revascularization of lower extremity injuries to prevent compartment syndrome, but the real-world impact is unknown. We assessed the association between fasciotomy and amputation and limb complications among lower extremitys with vascular injury. A retrospectively collected lower extremity injury database was queried for limbs undergoing attempted salvage with vascular procedure (2004-2012). Limbs were categorized as having undergone fasciotomy or not. Injury and treatment characteristics were collected, as were intervention timing data when available. The primary outcome measure was amputation. Multivariate models examined the impact of fasciotomy on limb outcomes. Inclusion criteria were met by 515 limbs, 335 (65%) with fasciotomy (median 7.7 h postinjury). Of 212 limbs, 174 (84%) with timing data had fasciotomy within 30 min of initial surgery. Compartment syndrome and suspicion of elevated pressure was documented in 127 limbs (25%; 122 had fasciotomy). Tourniquet and shunt use, fracture, multiple arterial and combined arteriovenous injuries, popliteal involvement, and graft reconstruction were more common in fasciotomy limbs. Isolated venous injury and vascular ligation were more common in nonfasciotomy limbs. Fasciotomy timing was not associated with amputation. Controlling for limb injury severity, fasciotomy was not associated with amputation but was associated with limb infection, motor dysfunction, and contracture. Sixty-three percent of fasciotomies were open for >7 d, and 43% had multiple closure procedures. Fasciotomy revision (17%) was not associated with increased amputation or complications. Fasciotomy after military lower extremity vascular injury is predominantly performed early, frequently without documented compartment pressure elevation. Early fasciotomy is generally performed in severely injured limbs with a subsequent high rate of limb complications.


Interaction of calponin with phospholipids.
The interaction between chicken gizzard calponin and phospholipids was examined by sedimentation assay and affinity chromatography. Calponin was sedimented with phosphatidylserine (PS) and phosphatidylinositol (PI) vesicles but not with phosphatidylcholine (PC) vesicles. The apparent Kd values of calponin to PS and PI were calculated to be 1.3 x 10(6) and 1.5 x 10(6) M-1, respectively. Domain mapping with chymotryptic digestion showed that the phospholipid-binding site resided within the N-terminal 22-kDa fragment, in which the bindings of actin, calmodulin, S100, and tropomyosin also occur. The amount of calponin bound to PS and PI vesicles decreased with increasing ionic strength or Ca2+ concentrations. The presence of MgCl2 was needed for the calponin-PS vesicle interaction. Calponin-binding proteins including actin, calmodulin, and S100 inhibited calponin binding to the phospholipid vesicles in a concentration-dependent manner, while tropomyosin had little effect on the binding. The inhibitory effects of calmodulin and S100 were found only in the presence of CaCl2. Neither caldesmon nor SM22 affected the binding.


Secondary angle closure caused by air migrating behind the pupil in descemet stripping endothelial keratoplasty.
To report secondary angle closure caused by air migrating behind the pupil in the context of intraocular pressure (IOP) elevation in the early postoperative period after Descemet stripping endothelial keratoplasty (DSEK). A retrospective case series was conducted on 100 consecutive DSEK cases from 90 patients undergoing DSEK because of corneal disease from Fuchs corneal dystrophy, pseudophakic bullous keratopathy, aphakic bullous keratopathy, and iridocorneal endothelial syndrome. Preoperative and postoperative slit-lamp examinations and IOP measurements were ascertained for all 100 eyes. Main outcome measures included preoperative and postoperative IOP. Thirteen of 100 eyes developed an IOP rise of greater than 30 mm Hg on the first postoperative day. Six of these 13 patients developed angle closure from air migrating posterior to the iris and causing iridocorneal adhesions. One of these 13 patients developed pupillary block from air anterior to iris. Six of 13 patients developed increased IOP without pupillary block or iridocorneal adhesions and had a history of preexisting primary or secondary glaucoma. A secondary angle closure associated with DSEK is reported with air migrating behind the iris, resulting in extensive iridocorneal adhesions. An acute increase in IOP after DSEK can also be induced by air anterior to the iris causing pupillary block. IOP spikes are much more common in the first few postoperative days after DSEK. Medical treatment can occasionally resolve air posterior to the iris, but if iridocorneal adhesions are extensive and persistent, air removal and angle reformation may be necessary.


Nodular amyloidoma and primary pulmonary lymphoma with amyloid production: a differential diagnostic problem.
Nodular amyloidomas (NA) of the lung are non-neoplastic inflammatory nodules containing eosinophilic amyloid deposits and a lymphoplasmacytic infiltrate. In some instances, the extensive amyloid deposits may obscure an underlying lymphoproliferative disorder. The histologic and immunohistologic features that discriminate these two differential diagnostic possibilities were studied in this series of six cases of NA and five cases of primary low-grade malignant lymphomas of lung with secondary amyloid deposits (ML). Two of lymphoma cases showed histopathologic and immunophenotypic features of B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma (B-cell CLL/SLL), and three cases were low-grade B-cell lymphoma derived from mucosa associated lymphoid tissue (MALT lymphoma). Key discriminating morphologic features between NA and ML included lymphatic tracking of the cellular infiltrate (3/5 ML; 1/6 NA), pleural infiltration (3/5 ML; 0/6 NA), sheet-like masses of plasma cells (5/5 ML; 0/6 NA) and reactive follicles (4/5 ML; 1/6 NA). Lesional circumscription, vascular and bronchial destruction, lymphoepithelial lesions, and granulomas were not helpful discriminators. Immunohistochemical features indicating a dominant CD20+, CD79a+ B-cell population (5/5 ML; 0/6 NA), light chain restriction (4/5 ML; 0/6 NA), and aberrant antigen expression of CD20/CD43 (2/5 ML; 0/6 NA) were helpful. Amyloid tumors with a reactive lymphoplasmacytic infiltrate can be separated from low-grade malignant lymphomas utilizing both histologic and immunohistochemical features.


Effect of chronic erythropoietin administration on plasma iron in newborn lambs.
Erythropoietin, the primary stimulator of erythropoiesis, represents an important potential therapy for the anemia of prematurity. Enhancement of the therapeutic benefit of recombinant human erythropoietin (rhEp) in very-low-birth-weight infants will require a better understanding of rhEp's pharmacodynamic effects including its interaction with iron in stimulating erythropoiesis. The purpose of this study was to determine the effects of chronic rhEp administration on plasma iron levels and hematopoiesis using a twin lamb model. Nine pairs of twin lambs in which one twin was randomized to receive rhEp, and the other saline, were studied during a 1-week baseline and a subsequent 4- to 5-week treatment period. The effects of therapy on plasma iron levels and erythropoiesis were measured by integrating the areas under the concentration-time curves (AUC) of the study variables. During the rhEp treatment period, significantly greater negative daily AUCs were observed in the rhEp-treated lambs for plasma iron concentration (p = 0.0008), while significantly greater positive daily AUCs were observed for hemoglobin concentration (p = 0.04) and reticulocyte count (p = 0.02). In the rhEp-treated group, pretreatment iron concentrations were directly associated with the magnitude of the iron response during treatment such that the greater the pretreatment iron, the greater the daily AUC below the plasma iron concentration-time plot (r = -0.66, p = 0.05). For the placebo-treated group, this association tended toward, but did not achieve, statistical significance (r = -0.52, p = n.s.). These observations suggest that treatment of rapidly growing newborn lambs with rhEp results in increased iron utilization due to increased erythropoiesis and depends on iron status at the initiation of rhEp treatment. Use of the term neonatal lamb model offers advantages over studies in human infants for more detailed or invasive examinations of the interaction of iron and rhEp treatment.


Hydroxychloroquine. Seven-year experience.
Ninety-nine patients were treated with hydroxychloroquine sulfate for connective tissue diseases during a seven year period. All patients received the medication for at least one year and most were treated with a daily dosage of 400 mg. Four patients experienced toxic effects, but no patient sustained permanent loss of visual acuity. Toxicity did not seem to represent a cumulative dosage effect. Funduscopic and visual field examinations at six-month intervals are sufficient to detect the occasional adverse reactions and prevent serious retinal toxic effects when a maximum daily dosage of 400 mg is maintained.


Quantized gravitational responses, the sign problem, and quantum complexity.
It is believed that not all quantum systems can be simulated efficiently using classical computational resources. This notion is supported by the fact that it is not known how to express the partition function in a sign-free manner in quantum Monte Carlo (QMC) simulations for a large number of important problems. The answer to the question-whether there is a fundamental obstruction to such a sign-free representation in generic quantum systems-remains unclear. Focusing on systems with bosonic degrees of freedom, we show that quantized gravitational responses appear as obstructions to local sign-free QMC. In condensed matter physics settings, these responses, such as thermal Hall conductance, are associated with fractional quantum Hall effects. We show that similar arguments also hold in the case of spontaneously broken time-reversal (TR) symmetry such as in the chiral phase of a perturbed quantum Kagome antiferromagnet. The connection between quantized gravitational responses and the sign problem is also manifested in certain vertex models, where TR symmetry is preserved.


A potent gelatinase inhibitor with anti-tumor-invasive activity and its metabolic disposition.
Metastatic tumors lead to more than 90% fatality. Despite the importance of invasiveness of tumors to poor disease outcome, no anti-invasive compounds have been commercialized. We describe herein the synthesis and evaluation of 4-(4-(thiiranylmethylsulfonyl)phenoxy)-phenyl methanesulfonate (compound 2) as a potent and selective inhibitor of gelatinases (matrix metalloproteinases-2 and -9), two enzymes implicated in invasiveness of tumors. It was demonstrated that compound 2 significantly attenuated the invasiveness of human fibrosarcoma cells (HT1080). The metabolism of compound 2 involved hydroxylation at the alpha-methylene, which generates sulfinic acid, thiirane ring-opening, followed by methylation and oxidation, and cysteine conjugation of both the thiirane and phenyl rings.


[Cytoflavin in the treatment of patients with chronic abacterial prostatitis and erectile dysfunction].
The problem of chronic prostatitis (CP) and erectile dysfunction (ED) involves not only their high prevalence, but also the insufficient effectiveness of their treatments. In this regard, there is need for administration the pathogenetic drugs with antihypoxic, antioxidant and neuroprotective effects and improving blood flow to the genitals. The study included 60 men with CP associated with ED, aged 22 to 60 years. Patients were randomized into 2 groups of 30 people. Patients of comparison group received baseline therapy (alpha1-adrenoblockers, non-specific anti-inflammatory drugs, digital prostate massage and vacuum phallostimulation). Antibiotics were applied on the basis of their potential effectiveness in chronic abacterial prostatitis. In addition to the above treatment, patients of main group received step-down therapy with cytoflavin (in infusion, then oral administration). Positive dynamics was noted in both groups of patients; however, according to the indicators of the severity of pain and dysuria, as well as quality of life, positive dynamics in the main group of patients was more significant. Similarly, the dynamics of objective criteria for inflammation in the prostate gland was more pronounced when using cytoflavin. After treatment, the rigid phase of erection during vacuum fallotest occurred within 2-3 min from the beginning of the procedure in 16 (53.3%) patients of main group and only in 9 (30%) patients of comparison group. During follow-up examination at 6 months after treatment, stable remission was found in 75% of patients of main group. Thus, the inclusion of cytoflavin in the scheme of complex treatment of patients with abacterial CP associated with ED is pathogenetically justified, makes it more efficient and provides good DFS.


Approximal caries increment in adolescents in a low caries prevalence area in Sweden after a 3.5-year school-based fluoride varnish programme with Bifluorid 12 and Duraphat.
To evaluate approximal caries increment among 12- to 16-year-olds in a low caries prevalence area in Sweden after a 3.5-year school-based fluoride (F) varnish programme with Bifluorid 12 and Duraphat. The design was a RCT study with 1365 adolescents, divided into following four groups: Group 1 Bifluorid 12 two applications/year; Group 2 Duraphat two applications/year; Group 3 Bifluorid 12 four applications/year and Group 4 no F varnish at school. 1143 children (84%) completed the study. Approximal caries was registered on bitewing radiographs. There were no statistically significant differences in caries prevalence among the groups either at baseline or after 3.5 years . The caries increment for Group 1 was 1.34 ± 2.99 (mean ± SD), 1.24 ± 2.84 for Group 2, 1.07 ± 2.66 for Group 3 and 1.25 ± 2.75 for Group 4, with no statically significant differences either between Bifluorid 12 and Duraphat with the same frequency of F varnish applications or between the F groups and the control group. In an area with low caries prevalence in Sweden, the supplementary caries-preventive effect of school-based F varnish applications, to regular use of F toothpaste at home and to regular caries prevention given at the Public Dental Clinics, appears to be nonsignificant regarding approximal caries increment.


Resveratrol Enhances Chemosensitivity in Mouse Melanoma Model Through Connexin 43 Upregulation.
Although current studies indicate that resveratrol exhibits potential antitumor activities, the precise mechanisms of its beneficial effects combined with chemotherapy are not fully understood. This work is warranted to elucidate the underlying mechanism of antitumor effects by the combination therapy of resveratrol and cisplatin. The presence of functional gap junctions is highly relevant for the success of chemotherapy. Gap junctions mediate cell communication by allowing the passage of molecules from one cell to another. Connexin (Cx) 43 is ubiquitous and reduced in a variety of tumor cells. Cx43 may influence the response of tumor cells to treatments by facilitating the passage of antitumor drugs or death signals between neighboring tumor cells. Following resveratrol treatment, dose-dependent upregulation of Cx43 expressions was observed. In addition, gap junction intercellular communication was increased. To study the mechanism underlying these resveratrol-induced Cx43 expressions, we found that resveratrol induced a significant increase in mitogen-activated protein kinases (MAPK) signaling pathways. The MAPK inhibitors significantly reduced the expression of Cx43 protein after resveratrol treatment. Specific knockdown of Cx43 resulted in a reduction of cell death after resveratrol and cisplatin treatment. Our results suggest that treatment of resveratrol in tumor leads to increase Cx43 gap junction communication and enhances the combination of resveratrol and cisplatin therapeutic effects. © 2014 Wiley Periodicals, Inc. Environ Toxicol 30: 877-886, 2015.


[The role and significance of the most important known cytokines in inflammatory bowel diseases].
Several theories have already been postulated in connection with the pathogenesis of inflammatory bowel diseases, yet none of them has been approved. Recently increasing attention has been payed to different cytokines, playing central role in the development of inflammatory processes. In the intestinal mucosa of patients suffering from inflammatory bowel diseases increasing amounts of interleukin-1 (IL-1), tumor necrosis factor (TNF) and platelet activating factor (PAF) could be measured. On the other hand, antiinflammatory cytokines seem to be ineffective, or being present in insufficient amount (IL-4 and IL-10 respectively). It is therefore probable, that altered ratios of cytokines, or pathologic regulation of their production lead to progression of inflammation in IBD. Influence of cytokine production may open new therapeutic approach, e.g. IL-10 enema proved to be effective in the treatment of some cases of steroid-resistant ulcerative colitis, while intravenous administration was useful in Crohn's disease. A brief, comprehensive review of our present knowledge about cytokines in IBD is given.


Activation of phosphorylase in frog muscle as determined by contractile activity.
The state of activation of phosphorylation in muscle has been reinvestigated by combining the extraction procedures of Danforth, Helmreich, and Cori with the low-temperature techniques of this laboratory. In resting frog muscle, the phosphorylase-alpha content is usually below detectability. Upon contractile activity in series of twitches, activation of phosphorylase beta to alpha took place, without activation of phosphorylase beta kinase as defined by the assay procedure. Two different experimental designs were used to examine the relation between phosphorylase activation and the myothermally determined energy turnover per twitch, and these showed, identically, that the enzyme activation is proportional to the energy per twitch.


Missense mutations in the BCS1L gene as a cause of the Björnstad syndrome.
The Björnstad syndrome, an autosomal recessive disorder associated with sensorineural hearing loss and pili torti, is caused by mutation of a previously unidentified gene on chromosome 2q34-36. Refined genetic mapping and DNA sequencing of 44 genes between D2S2210 and D2S2244 revealed BCS1L mutations. Functional analyses elucidated how BCS1L mutations cause the Björnstad syndrome. BCS1L encodes a member of the AAA family of ATPases that is necessary for the assembly of complex III in the mitochondria. In addition to the Björnstad syndrome, BCS1L mutations cause complex III deficiency and the GRACILE syndrome, which in neonates are lethal conditions that have multisystem and neurologic manifestations typifying severe mitochondrial disorders. Patients with the Björnstad syndrome have mutations that alter residues involved in protein-protein interactions, whereas mutations in patients with complex III deficiency alter ATP-binding residues, as deduced from the crystal structure of a related AAA-family ATPase. Biochemical studies provided evidence to support this model: complex III deficiency mutations prevented ATP-dependent assembly of BCS1L-associated complexes. All mutant BCS1L proteins disrupted the assembly of complex III, reduced the activity of the mitochondrial electron-transport chain, and increased the production of reactive oxygen species. However, only mutations associated with complex III deficiency increased mitochondrial content, which further increased the production of reactive oxygen species. BCS1L mutations cause disease phenotypes ranging from highly restricted pili torti and sensorineural hearing loss (the Björnstad syndrome) to profound multisystem organ failure (complex III deficiency and the GRACILE syndrome). All BCS1L mutations disrupted the assembly of mitochondrial respirasomes (the basic unit for respiration in human mitochondria), but the clinical expression of the mutations was correlated with the production of reactive oxygen species. Mutations that cause the Björnstad syndrome illustrate the exquisite sensitivity of ear and hair tissues to mitochondrial function, particularly to the production of reactive oxygen species.


Diurnal variation of alpha 1-acid glycoprotein concentration in normal volunteers.
The serum concentration of alpha 1-acid glycoprotein (AAG) was determined hourly for 25 h in male and female volunteers. Subjects were carefully selected to control for age, weight, health status, smoking history, medication use, and, for females, estrus cycle and oral contraceptive use. Studies were performed under conditions closely controlled for sleep, daytime environment, and diet. All mean concentrations were within the range reported as normal, but the deviation around individual means varied from 6.4 to 48.9%. Males had higher mean AAG concentrations than females, 59.9 versus 52.4 mg/dL, but the difference was not statistically significant. Studies which compare changes in AAG concentration or the degree of basic drug protein binding over time within individuals or between groups should consider intrasubject variability in AAG for accurate interpretation of results.


Properties of human blood monocytes. II. Monocytes from healthy adults are highly heterogeneous within and among individuals.
Human blood monocytes are known to include subsets defined by the expression of CD14 and CD16 but otherwise are often assumed to be relatively homogeneous. However, we had observed additional heterogeneity that led us to a more extensive examination of monocytes. Blood samples from 200 healthy adults without known immunological abnormalities were examined by analysis with a hematology analyzer and by flow cytometry (FCM) to determine leukocyte differential counts, to identify subsets and to measure expression of monocyte-associated molecules. The estimated cell counts of monocytes, neutrophils, total lymphocytes, and T cells all varied to a similar extent, that is, ±30-35%. The fractions of monocyte subsets defined by CD14 and CD16 or by CD163 expression also varied among individuals. FCM examinations showed that all the monocyte-associated molecules that were examined varied in expression in this increasing order-CD244, CD4, CD38, CD91, CD11b, toll-like receptor 2 (TLR2), TIA-1, CD14 (on CD14(Br+) cells), CD86, CD80, HLA-DQ, CD33, and HLA-DR. Human blood monocytes are heterogeneous among healthy adults with respect to cell counts, subsets, and the levels of expression of monocyte-associated molecules. An increase in the "non-classical" (CD14(Lo/Neg) /CD16(+) ) monocyte subset or in the expression of CD11b or TLR2 have known diagnostic/prognostic implications. CD244 and CD4 have well-defined functions on lymphocytes but perform unknown activities on monocytes although their expression appears more narrowly controlled. Together, these data suggest that monocytes should be more extensively examined in both clinical and basic contexts.


Metabolic Profiling of Glucose-Fed Metabolically Active Resting Zymomonas mobilis Strains.
Zymomonas mobilis is the most efficient bacterial ethanol producer and its physiology is potentially applicable to industrial-scale bioethanol production. However, compared to other industrially important microorganisms, the Z. mobilis metabolome and adaptation to various nutritional and genetic perturbations have been poorly characterized. For rational metabolic engineering, it is essential to understand how central metabolism and intracellular redox balance are maintained in Z. mobilis under various conditions. In this study, we applied quantitative mass spectrometry-based metabolomics to explore how glucose-fed non-growing Z. mobilis Zm6 cells metabolically adapt to change of oxygen availability. Mutants partially impaired in ethanol synthesis (Zm6 adhB) or oxidative stress response (Zm6 cat) were also examined. Distinct patterns of adaptation of central metabolite pools due to the change in cultivation condition and between the mutants and Zm6 reference strain were observed. Decreased NADH/NAD ratio under aerobic incubation corresponded to higher concentrations of the phosphorylated glycolytic intermediates, in accordance with predictions of the kinetic model of Entner-Doudoroff pathway. The effects on the metabolite pools of aerobic to anaerobic transition were similar in the mutants, yet less pronounced. The present data on metabolic plasticity of non-growing Z. mobilis cells will facilitate the further metabolic engineering of the respective strains and their application as biocatalysts.


Fluid-structure interaction simulations of venous valves: A monolithic ALE method for large structural displacements.
Venous valves are bicuspidal valves that ensure that blood in veins only flows back to the heart. To prevent retrograde blood flow, the two intraluminal leaflets meet in the center of the vein and occlude the vessel. In fluid-structure interaction (FSI) simulations of venous valves, the large structural displacements may lead to mesh deteriorations and entanglements, causing instabilities of the solver and, consequently, the numerical solution to diverge. In this paper, we propose an arbitrary Lagrangian-Eulerian (ALE) scheme for FSI simulations designed to solve these instabilities. A monolithic formulation for the FSI problem is considered, and due to the complexity of the operators, the exact Jacobian matrix is evaluated using automatic differentiation. The method relies on the introduction of a staggered in time velocity to improve stability, and on fictitious springs to model the contact force of the valve leaflets. Because the large structural displacements may compromise the quality of the fluid mesh as well, a smoother fluid displacement, obtained with the introduction of a scaling factor that measures the distance of a fluid element from the valve leaflet tip, guarantees that there are no mesh entanglements in the fluid domain. To further improve stability, a streamline upwind Petrov-Galerkin (SUPG) method is employed. The proposed ALE scheme is applied to a two-dimensional (2D) model of a venous valve. The presented simulations show that the proposed method deals well with the large structural displacements of the problem, allowing a reconstruction of the valve behavior in both the opening and closing phase.


[Humoral antibodies to Epstein-Barr virus in infectious mononucleosis].
A high level of humoral antibody to the capsid antigen of Epstein-Barr virus was found in sera from patients with infectious mononucleosis. For detection of antibody, the method of indirect immunofluorescence in fixed P3HEHR cells of Burkitt's lymphoma was used. Sera from normal subjects were also shown to contain antibody to the capsid antigen of Epstein-Barr virus, but the titers were 5--10 times lower than in the sera from infectious mononucleosis patients. The occurrance of high titers of antibody to EBV in the sera from patients with infectious mononucleosis is diagnostic for IM and may be used for practical purposes.


Proteins from human cerebrospinal fluid: binding with nucleic acids.
Cerebrospinal fluid (CSF) contains two groups of proteins that bind tightly to DNA and to polyriboguanylic acid, respectively. In certain diseases the amounts of a given nucleic acid bound by a constant volume of CSF may increase, while in others the amount of such proteins may be reduced. Binding of polyriboguanylic acid increased in CSF samples from patients with brain tumors, stroke, multiple sclerosis, and communicating hydrocephalus, but it significantly decreased in CSF samples from patients with obstructive hydrocephalus. These increases may or may not be proportional to the rise in total CSF proteins characteristic for these diseases. Elevated binding of DNA was observed in samples from patients with hydrocephalus, epilepsy, and cortical atrophy. The technique described may be applicable to the diagnosis of a variety of diseases of the central nervous system.


Australia antigen in chronic hepatitis in Australia.
Of 53 cases of active chronic liver disease two were found to be carriers of Australia antigen Au (1)-an elderly woman with typical lupoid hepatitis and an elderly mortuary attendant with serologically atypical active chronic hepatitis. Au (1) was detected also in the serum of 7 out of 20 patients with clinically atypical acute hepatitis-one was an elderly Italian woman, one had hepatitis in the puerperium, and five had a history of transfusion or inoculation. The antigen was not found in 20 typical cases of infectious hepatitis in young people in 86 patients with other diseases. Antibody to Au (1) was present in only 2 out of 102 patients who had received numerous transfusions.We conclude, firstly, that Au (1) antigen is rare in white Australians (in keeping with the low incidence of serum hepatitis in Australia), and, secondly, that Au (1) positivity in hepatitis patients is associated with transfusions and with older age. We suggest that active chronic hepatitis and lupoid hepatitis may follow infection of susceptible individuals with Au (1)-positive hepatitis virus, but persistence of the virus in high titre does not appear to be necessary for chronicity of the disease.


Exon 6 variants carried on systemic lupus erythematosus (SLE) risk haplotypes modulate IRF5 function.
Interferon regulatory factor 5 (IRF5) regulates innate immune responses to viral infection. IRF5 genetic variants have been shown to be strongly associated with risk for systemic lupus erythematosus (SLE). Functional roles of IRF5 exon 6 structural variants that occur as part of a SLE risk-associated haplotype, including a 30-bp in/del (in/del-10) and a 48-bp splice-site variant (SV-16), have not been established. In this study, we used IRF5-deficient cells overexpressing human IRF5 (hIRF5) variants to investigate the roles of exon 6 in/del-10 and SV-16 in regulation of the apoptosis response, nuclear translocation, and ability to transactivate IRF5 responsive cytokines. We found that expression of IRF5 isoforms including either SV-16 or in/del-10 confers ability of IRF5 to impair the apoptotic response and correlates with reduced capacity for IRF5 nuclear translocation in MEFs after a DNA-damaging stimulus treatment. Interestingly, the presence or absence of both SV-16 and in/del-10 results in abrogation of both the anti-apoptotic and enhanced nuclear translocation effects of IRF5 expression. Only cells expressing IRF5 bearing SV-16 show increased IL-6 production upon lipopolysaccharide stimulation. MEFs expressing hIRF5 variants containing in/del-10 showed no significant difference from the control; however, cells carrying hIRF5 lacking both SV-16 and in/del-10 showed reduced IL-6 production. Our overall findings suggest that exon 6 SV-16 is more potent than in/del-10 for IRF5-driven resistance to apoptosis and promotion of cytokine production; however, in/del-10 co-expression can neutralize these effects of SV-16.


Objective indicators of physical activity and sedentary time and associations with subjective well-being in adults aged 70 and over.
This study explored the associations of the volume and intensity of physical activity and the volume of sedentary time with subjective well-being in a diverse group of 228 older adults in the UK (111 female, mean age 78.2 years (SD 5.8)). Physical activity (PA) and sedentary behaviour were assessed by accelerometry deriving mean steps per day, mean moderate/vigorous PA minutes per hour (MVPA min · h(-1)) and minutes of sedentary time per hour (ST min · h(-1)). Lower limb function was assessed by the Short Physical Performance Battery. Subjective well-being was assessed using the SF-12 health status scale, the Ageing Well Profile and the Satisfaction with Life Scale. Linear regressions were used to investigate associations between the independent variables which included physical activity (steps and MVPA), sedentary time, participant characteristics (gender, age, BMI, education, number of medical conditions), and lower limb function and dependent variables which included mental and physical well-being. Steps, MVPA and lower limb function were independently and moderately positively associated with perceived physical well-being but relationships with mental well-being variables were weak. No significant associations between sedentary behaviours and well-being were observed. The association between objectively evaluated physical activity and function and subjective evaluations of physical well-being suggest that improving perceptions of physical health and function may provide an important target for physical activity programmes. This in turn may drive further activity participation.


Questionnaire-based case finding of celiac disease in a population of 8- to 9-year-old children.
Antibody screenings and diagnosis of celiac disease (CD) among children with type 1 diabetes have suggested that a considerable proportion of children with CD may, in fact, have preclinical (undiagnosed) symptoms. We aimed to test if a questionnaire would lead to significant case finding in an unselected population of 8- to 9-year-old children. The study population included 9880 children aged 8 to 9 years. Before the study, 13 children from the study population were known to have CD. We developed a questionnaire on the basis of 5 simple items suggestive of CD and mailed the questionnaire to the families of all children in the study population who resided in the County of Funen, Denmark. In total, 7029 respondents returned the questionnaire (70%); among them, 2835 children had 1 or more symptoms. These children were invited for a blood test to determine their human serum immunoglobulin A (IgA) anti-tissue transglutaminase antibody (anti-tTG) levels. Of the 1720 children who were tested for the human serum IgA anti-tTG, 24 participants had a positive result (range: 20 to >150 U). Seventeen of these children underwent an upper endoscopy procedure. Fourteen children had histologic signs of CD (Marsh classification stage III). Fourteen children met the diagnostic criteria for CD. The prevalence proportion of patients who were newly diagnosed with CD was 0.14% (95% CI: 0.08-0.24) (14 of 9967), and the estimate of the minimum total prevalence proportion of children with CD was 0.27% (95% CI: 0.18-0.39) (27 of 9980). The maximal prevalence proportion of patients with newly diagnosed CD was 1.22% (95% CI: 0.76-1.90) (21 of 1720), including those participants who had a positive anti-tTG result but not a final diagnosis of CD. The ratio of known to minimally symptomatic CD was approximately 1:1. A number of preclinical and low-grade symptomatic patients with CD may be identified by their responses to a mailed questionnaire.


Pediatric Heart Transplantation: Transitioning to Adult Care (TRANSIT): Baseline Findings.
Young adult solid organ transplant recipients who transfer from pediatric to adult care experience poor outcomes related to decreased adherence to the medical regimen. Our pilot trial for young adults who had heart transplant (HT) who transfer to adult care tests an intervention focused on increasing HT knowledge, self-management and self-advocacy skills, and enhancing support, as compared to usual care. We report baseline findings between groups regarding (1) patient-level outcomes and (2) components of the intervention. From 3/14 to 9/16, 88 subjects enrolled and randomized to intervention (n = 43) or usual care (n = 45) at six pediatric HT centers. Patient self-report questionnaires and medical records data were collected at baseline, and 3 and 6 months after transfer. For this report, baseline findings (at enrollment and prior to transfer to adult care) were analyzed using Chi-square and t-tests. Level of significance was p < 0.05. Baseline demographics were similar in the intervention and usual care arms: age 21.3 ± 3.2 vs 21.5 ± 3.3 years and female 44% vs 49%, respectively. At baseline, there were no differences between intervention and usual care for use of tacrolimus (70 vs 62%); tacrolimus level (mean ± SD = 6.5 ± 2.3 ng/ml vs 5.6 ± 2.3 ng/ml); average of the within patient standard deviation of the baseline mean tacrolimus levels (1.6 vs 1.3); and adherence to the medical regimen [3.6 ± 0.4 vs 3.5 ± 0.5 (1 = hardly ever to 4 = all of the time)], respectively. At baseline, both groups had a modest amount of HT knowledge, were learning self-management and self-advocacy, and perceived they were adequately supported. Baseline findings indicate that transitioning HT recipients lack essential knowledge about HT and have incomplete self-management and self-advocacy skills.


Infection after rotator cuff repair.
Sixteen patients (15 men and 1 woman) were treated for infection complicating rotator cuff repair during the period 1975 through 1994. Eight of the 16 patients had their initial procedure performed elsewhere. The remaining 8 procedures were performed at our institution with the known incidence of this complication being 0.27%. In addition to intravenous antibiotic therapy, an average of 3.5 (range 2 to 8) operative procedures were required to eradicate the infections. Micro-organisms cultured were Propionibacter in 6, coagulase negative Staphylococcus in 4, Staphylococcus aureus in 4, Peptostreptococcus magnus in 1, and both Propionibacter and coagulase-negative Staphylococcus in 1. The deltoid was restored in all patients; the rotator cuff was reparable in 11. In the 12 shoulders with greater than 1 year of follow-up (average 51 months, range 14 to 165 months), active elevation averaged 110 degrees and external rotation 50 degrees. Four patients had no pain, 4 had minimal pain, and the remaining 4 had moderate pain. Satisfactory final results, which were determined by the patients' opinion or with the use of either the University of California, Los Angeles score or the modified Neer system, were obtained in 5 (42%) of the shoulders.


Action potentials in normal and Shaker mutant Drosophila.
Intracellular microelectrode recordings from the cervical giant fiber of normal Drosophila show a characteristic action potential waveform for this identified neuron. The action potential has a rapid initial spike followed by a prominent depolarizing afterpotential. Pharmacological experiments suggest that the giant fiber action potential depends on inward currents carried by Na+ and outward currents carried by K+. Abnormal action potentials are seen in Shaker (Sh) mutant Drosophila. This study compares the effects of six Sh alleles. In each case, abnormalities are limited to action potential repolarization. There are, however, allelic differences. Five alleles cause delayed repolarization and increased action potential durations. Going from most to least extreme, these alleles are: Sh102 greater than ShKS133 greater than ShM greater than ShE62 greater than ShrKO120. Compared to normal action potentials, durations in the extreme mutants are longer by an order of magnitude or more. One mutant allele, Sh5 appears to cause an incompletely repolarized action potential, rather than a repolarization delay.


Biocide activity of Annona coriacea seeds extract on Rhodnius neglectus (Hemiptera: Reduviidae).
The use of synthetic insecticides for insect control may lead to different kind of problems, such as vector resistance to insecticides. To avoid these problems, a new research area to study botanical products as possible disease vectors controls, has become a feasible alternative. The purpose of this research was to evaluate the biocide activity of the ethanol extract of seeds of Annona coriacea on Rhodnius neglectus (Chagas disease vector) nymphs and adults. For this, different concentrations extracts were evaluated: 25, 50, 100 and 200mg/ mL, and water in DMSO (20%) was used as control. The experimental design was completely randomized and we conducted the bioassay with nymphs and adults, with 10 nymphs and 10 adults (five males and five females) per treatment. Extract action was evaluated in both bioassays, in order to identify possible effects of mortality and life cycle interruption of nymphs and adults during a 28-day-period. The results obtained showed that the extract of A. coriacea was able to disrupt the development of nymphs and adults of R. neglectus, with a mortality rate of more than 90%, 36% and 100%, at the highest concentrations, respectively. There was also molting inhibition in nymphs, lower reproductive capacity in females, feeding deterrence and morphological changes in nymphs and adults. We concluded that the extract of A. coriacea has insecticide action on nymphs and adults of R. neglectus.


[NMR-tomography of the human eye].
The NMR relaxation data of tissue are the basis of an appropriate adjustment of NMR imaging parameters. The relaxation data of ocular tissues are presented, and the relaxation behaviour of the eye is discussed. NMR imaging of eyes with intraocular silicone and pseudophakia are shown. Up to the present time, neuritis of the optic nerve is not detectable by NMR tomography whereas intraocular melanoma are clearly distinguishable.


Comparison of accuracy and reliability of CBCT and 3D laser scanner in the volumetric assessment of the root canal space.
To compare the accuracy and reliability of cone-beam computed tomography (CBCT) and laser scanner in measuring minor volume changes such as the root canal space. 35 maxillary incisors were endodontically prepared. A dimensionally stable silicone material was injected into the root canal space and scanned with CBCT. The root canal volume was measured using Romexis 3.0.1 R software. Replicas were carefully removed from the teeth and scanned using an extraoral laser scanner. These images were exported to the Rhinoceros software for volume measurement. The volume of each replica was also assessed using the gravimetric method. To determine the accuracy, the volume obtained from both devices was compared with the gravimetric method. Statistical analysis was done using a paired t-test. The reliability was assessed using the intraclass correlation coefficient. There was no statistically significant difference between the mean volume of CBCT 27.04 ± 7.25 mm³ and the mean volume of the gravimetric method 27.87 ± 7.17 mm³ (P< 0.05). A statistically significant difference was seen with the laser scanner at 25.31 ± 6.89 mm³ and the gravimetric method at 27.87 ± 7.17 mm³ (P< 0.05). CBCT showed a good degree of agreement (ICC 0.899), while the laser scanner showed a moderate degree of agreement (ICC 0.644) with the gravimetric method. CBCT proved accurate and reliable in measuring minor volumes like the root canal space, ideally in the range of 20-25 mm³. The laser scanner presented acceptable reliability. The laboratory data showed satisfactory outcomes, providing an evidence-based approach and potentially motivating clinicians to integrate cone-beam computed tomography for volume analysis into clinical practice. The accuracy and reliability of laser scanners for small-volume analysis have not previously been evaluated. Consequently, the findings from this study warrant further clinical investigations.


Assessing an Epidemic: Utility of the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition Level 2 Substance Use Screener in Adult Psychiatric Inpatients.
Inpatient psychiatric hospitals provide an important point of care for assessing and stabilizing substance use and for facilitating linkage to appropriate treatment. Toxicology screening provides a key measure of substance use yet may miss many cases of substance use because of variable windows of detection and the limited scope of substances assessed. This study assesses the utility of the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) Level 2 Substance Use screener as a supplemental tool for identifying substance use by self-report within an inpatient psychiatric hospital setting. From a larger sample of 97 adult psychiatric inpatients, 60 who underwent drug toxicology testing and completed the DSM-5 screener were assessed. We examined the sensitivity and specificity of the self-report screener in comparison with drug toxicology test results collected by chart review. Sensitivity of the DSM-5 screener varied across substances assessed: The self-report measure identified 100% of individuals who tested positive for opioid use, 83% who tested positive for cannabis use, 50% who tested positive for cocaine use, and 37% who tested positive for benzodiazepine use. The self-report measure also identified 27 instances among 60 participants in which substance use identified by self-report was not detected by toxicology testing. The brief and easily administered DSM-5 Level 2 Substance Use screener shows promise for improving identification of substance use in an inpatient psychiatric hospital setting. This measure may also provide psychiatric inpatient nursing staff with a means of working collaboratively with patients to assess substance use and coordinate appropriate treatment plans.


Effect of mutant p27(kipl) gene on human cholangiocarcinoma cell line, QBC(939).
To investigate the effects of exogenously mutated p27(kipl) (p27) on proliferation and apoptosis of human cholangiocarcinoma cell line, QBC(939)in vivo. Adenviral vectors were used to transfect mutated p27 cDNA into human QBC(939)cell line. Expression of p27 was detected by RT-PCR. Western blot. Cell growth, morphological change, cell cycle, apoptosis and cloning formation were determined by MTT assay and flow cytometry. The expression of p27 protein and mRNA was increased significantly in QBC(939) cell line transfected with Ad-p27mt. The transfer of Ad-p27mt could significantly inhibit the growth of QBC(939) cells, decrease the cloning formation rate and induce apoptosis. p27 over expression caused cell cycle arrest at G(0)/G(1)phase 72 h after infection with Ad-p27mt. p27 may cause cell cycle arrest at G(0)/G(1)phase and subsequently lead to apoptosis. Recombinant adenovirus expressing mutant p27 may be potentially useful in gene therapy for cholangiocarcinoma.


High ecological and human health risks from microcystins in vegetable fields in southern China.
Frequent cyanobacterial blooms in the eutrophic waters produce a variety of toxins such as the monocyclic heptapeptide microcystins, greatly harming aquatic ecosystems and human health. However, little information of microcystins in agricultural fields is known. This field study of three common microcystin variants (MC-LR, MC-RR, and MC-YR) in vegetables (n = 161), soils (n = 161) and irrigation water samples (n = 23) collected from southern China regions affected by cyanobacteria blooms, shows their prevalence with total concentrations up to 514 μg/L water, 187 μg/kg soil (dry weight) and 382 μg/kg vegetable (fresh weight). MC-RR was the primary variant in all types of samples, accounting for 51.3-100% of total microcystin concentrations. Significant concentration-dependent correlations (p < 0.05) demonstrated that microcystin-contained irrigation waters were the major source of microcystin accumulation in both vegetables and soils. Meanwhile, intracellular-microcystins in irrigation water was found to play an important role in microcystins bioaccumulation in vegetables for the first time. Most vegetable samples (≥60%), particularly celery posed moderate or high human health risk via diet based on toxicity equivalents of the microcystins and reference dose for MC-LR (0.04 μg/kg/d), showing high food safety hidden dangers. Soil microcystins, especially MC-RR in 46.4-88.3% of soils could pose high ecological risks. This study highlights the potential high ecological and human health risks of microcystins in the real soil-vegetable systems of areas affected by cyanobacteria blooms, implying the profound significance and urgent need of investigation on microcystins in terrestrial ecosystems.


Ergometric score systems after myocardial infarction: prognostic performance of the Duke Treadmill Score, Veterans Administration Medical Center Score, and of a novel score system, GISSI-2 Index, in a cohort of survivors of acute myocardial infarction.
The aims of the study were to evaluate the performance of the Duke Treadmill Score (DTS) and the Veteran Affairs Medical Center Score (VAMCS) in predicting 6-month death in GISSI-2 study survivors of acute myocardial infarction treated with thrombolytic agents, and to develop a simple predictive scoring system from the same database. Patients of the GISSI-2 study (n = 6251) performed a maximal symptom-limited exercise test 1 month after myocardial infarction. We calculated for each patient the DTS and the VAMCS. Based on the coefficients of a multivariate analysis of our database, we developed a simple predictive scoring system and performed an internal validation. The prognostic value of each scoring system was assessed by multivariate analysis. Six-month mortality rates in the subgroups of each scoring system were as follows: DTS: low risk 0.6%, moderate risk 1.8%, high risk 3.4% (P < or =.0001); VAMCS: low risk 0.6%, moderate risk 1.9%, high risk 4.7% (P < or =.0001); GISSI-2 Index: low risk 0.5%, moderate risk 1.9%, high risk 6.1% (P < or =.0001). The results of multivariate analysis (relative risk [RR] and 95% CI) were as follows: DTS: moderate risk 2.50 (1.47-12.59), high risk 5.13 (3.61-15.55); VAMCS: moderate risk 2.65 (1.53-4.59), high risk 5.97 (3.10-11.49); GISSI-2 Index: moderate risk 3.16 (1.81-5.52), high risk 8.65 (4.36-17.18). The use of ergometric-derived prognostic score systems in a population of survivors of acute myocardial infarction treated with thrombolytic drugs distinguishes subgroups at different risks of death and allows an appropriate recourse to more costly procedures.


In vivo colocalization of xyloglucan endotransglycosylase activity and its donor substrate in the elongation zone of Arabidopsis roots.
We have developed a method for the colocalization of xyloglucan endotransglycosylase (XET) activity and the donor substrates to which it has access in situ and in vivo. Sulforhodamine conjugates of xyloglucan oligosaccharides (XGO-SRs), infiltrated into the tissue, act as acceptor substrate for the enzyme; endogenous xyloglucan acts as donor substrate. Incorporation of the XGO-SRs into polymeric products in the cell wall yields an orange fluorescence indicative of the simultaneous colocalization, in the same compartment, of active XET and donor xyloglucan chains. The method is specific for XET, as shown by competition experiments with nonfluorescent acceptor oligosaccharides, by negligible reaction with cello-oligosaccharide-SR conjugates that are not XET acceptor substrates, by heat lability, and by pH optimum. Thin-layer chromatographic analysis of remaining unincorporated XGO-SRs showed that these substrates are not extensively hydrolyzed during the assays. A characteristic distribution pattern was found in Arabidopsis and tobacco roots: in both species, fluorescence was most prominent in the cell elongation zone of the root. Proposed roles of XET that include cell wall loosening and integration of newly synthesized xyloglucans could thus be supported.


Patients' and practice nurses' perceptions of depression in patients with type 2 diabetes and/or coronary heart disease screened for subthreshold depression.
Comorbid depression is common in patients with type 2 diabetes (DM2) and/or coronary heart disease (CHD) and is associated with poor quality of life and adverse health outcomes. However, little is known about patients' and practice nurses' (PNs) perceptions of depression. Tailoring care to these perceptions may affect depression detection and patient engagement with treatment and prevention programs. This study aimed to explore patients' and PNs' perceptions of depression in patients with DM2/CHD screened for subthreshold depression. A qualitative study was conducted as part of a Dutch stepped-care prevention project. Using a purposive sampling strategy, data were collected through semi-structured interviews with 15 patients and 9 PNs. After consent, all interviews were recorded, transcribed verbatim and analyzed independently by two researchers with Atlas.ti.5.7.1 software. The patient and PN datasets were inspected for commonalities using a constant comparative method, from which a final thematic framework was generated. Main themes were: illness perception, need for care and causes of depression. Patients generally considered themselves at least mildly depressed, but perceived severity levels were not always congruent with Patient Health Questionnaire 9 scores at inclusion. Initially recognizing or naming their mental state as a (subthreshold) depression was difficult for some. Having trouble sleeping was frequently experienced as the most burdensome symptom. Most experienced a need for care; psycho-educational advice and talking therapy were preferred. Perceived symptom severity corresponded with perceived need for care, but did not necessarily match help-seeking behaviour. Main named barriers to help-seeking were experienced stigma and lack of awareness of depression and mental health care possibilities. PNs frequently perceived patients as not depressed and with minimal need for specific care except for attention. Participants pointed to a mix of causes of depression, most related to negative life events and circumstances and perceived indirect links with DM2/CHD. Data of the interviewed patients and PNs suggest that they have different perceptions about (subthreshold) depressive illness and the need for care, although views on its causes seem to overlap more.