Latent autoimmune diabetes mellitus in children (LADC) with autoimmune thyroiditis and Celiac disease.
Latent autoimmune diabetes mellitus in adults (LADA) is characterized by clinical presentation as type 2 diabetes mellitus after 25 years of age, initial control achieved with oral hypoglycemic agents for at least 6 months, presence of autoantibodies and some immunogenetic features of type 1 diabetes mellitus. An 8.3 year-old girl was referred to our pediatric endocrinology department because of incidental glucosuria. She did not complain of polyuria, polydipsia, or weight loss. Her body mass index (BMI) was at the 80th percentile. Fasting glucose was 126 mg/dl, and OGTT glucose level at 120 min was 307 mg/dl. Although C-peptide levels were normal, her first phase insulin response (FIR) was lower than the 1st percentile. Anti-insulin antibody (AIA), islet cell antibody (ICA), and anti-glutamic acid decarboxylase (antiGAD) were negative. According to the clinical and laboratory findings, she was diagnosed as having type 2 diabetes mellitus. She was started with oral anti-diabetic treatment for a period of 1 year. Insulin had to be initiated for worsening of HbA1c levels. In the fourth year of follow-up, she was admitted to our hospital with diabetic ketoacidosis although she was on an intensive insulin regimen. At this time, C-peptide levels were low, antiGAD and AIA were positive with HLA DR3/DQ2 haplotype. In addition, her thyroid peroxidase antibody and endomysium antibody were found to be high at follow-up. Small intestinal biopsy revealed celiac disease. This patient may represent the first case of latent autoimmune diabetes mellitus in children (LADC) with autoimmune thyroiditis and celiac disease.


Theoretical study of thermal spin transition between the singlet state and the quintet state in the [Fe(2-picolylamine)(3)](2+) spin crossover system.
The spin transition between the low-spin singlet state and the high-spin quintet state in the [Fe(2-pic)(3)](2+) (2-pic: 2-picolylamine) complex is studied by using density functional theory (DFT) calculations. After careful comparison of density functionals BLYP, B3LYP, and B3LYP* (which has 15% Hartree-Fock exchange compared with 20% for B3LYP), we concluded that the spin-state splitting can be accurately reproduced by using the B3LYP* functional. The potential energy surfaces along minimum energy pathways of the three spin states were calculated at the B3LYP*/6-311+G** level of theory to find minimum energy crossing points (MECPs). The MECPs between the singlet and quintet states (SQ(M)) were found (E(SQ) = 6.8 kcal/mol), as well as the MECPs between the triplet and singlet states (ST(M), E(ST) = 12.9 kcal/mol) and the triplet and quintet states (TQ(M), E(TQ) = 12.8 kcal/mol). Although the distortion leading to SQ(M) from the singlet equilibrium geometry is mainly a symmetric expansion of the Fe-N bonds, the distortions leading to ST(M) and SQ(M) are asymmetric. Normal mode analysis demonstrates that these geometrical distortions contain a combination of several low-frequency normal modes, and therefore, these modes play a significant role in the intersystem crossing via the crossing seam.


Optimal trajectory for the occipital condylar screw.
Evaluation by simulation of screw insertion using fine-cut computed tomographic (CT) scans and screw trajectory software. To evaluate the feasibility and optimal trajectory of the occipital condylar screw. To the best of our knowledge, no large series examining the feasibility and optimal trajectory of occipital condylar screws have been published. We simulated unicortical placement of a 4 × 18-mm screw using 1-mm sliced CT scans and 3-dimensional screw trajectory software in 314 occipital condyles of 157 patients. With the screw tip directed toward a point just below the tip of the basion on lateral fluoroscopy, 3 entry points were compared in view of success rate of screw placement, safe range of medial angulation in an axial plane, and maximum screw length. The lateral entry point had the highest success rate (93.0%) of screw placement, mean safe range of the medial angulation (10.9°), and maximum screw length (20.7 mm), followed by the middle (92.0%, 10.8°, and 20.6 mm, respectively) and medial (74.2%, 7.7°, and 19.7 mm, respectively) entry points. While the lateral and the middle entry points did not have any statistically significant difference in the 3 parameters, they were significantly better than the medial entry point. All 3 entry points had highly variable ranges of safe medial angulation, making it difficult to provide a single recommended value. Screw placement is feasible in up to 93% of the occipital condyles. The lateral and the middle entry points are significantly better than the medial entry point. Selection between the middle and the lateral entry points should be individualized taking into account local anatomic variation. Because the medial angulation of these screws is highly variable, preoperative 3-dimensional CT evaluation and possibly even intraoperative navigation may be required.


SpoT Induces Intracellular Salmonella Virulence Programs in the Phagosome.
Guanosine tetraphosphate (ppGpp) and guanosine pentaphosphate (pppGpp), together named (p)ppGpp, regulate diverse aspects of Salmonella pathogenesis, including synthesis of nutrients, resistance to inflammatory mediators, and expression of secretion systems. In Salmonella, these nucleotide alarmones are generated by the synthetase activities of RelA and SpoT proteins. In addition, the (p)ppGpp hydrolase activity of the bifunctional SpoT protein is essential to preserve cell viability. The contribution of SpoT to physiology and pathogenesis has proven elusive in organisms such as Salmonella, because the hydrolytic activity of this RelA and SpoT homologue (RSH) is vital to prevent inhibitory effects of (p)ppGpp produced by a functional RelA. Here, we describe the biochemical and functional characterization of a spoT-Δctd mutant Salmonella strain encoding a SpoT protein that lacks the C-terminal regulatory elements collectively referred to as "ctd." Salmonella expressing the spoT-Δctd variant hydrolyzes (p)ppGpp with similar kinetics to those of wild-type bacteria, but it is defective at synthesizing (p)ppGpp in response to acidic pH. Salmonella spoT-Δctd mutants have virtually normal adaptations to nutritional, nitrosative, and oxidative stresses, but poorly induce metal cation uptake systems and Salmonella pathogenicity island 2 (SPI-2) genes in response to the acidic pH of the phagosome. Importantly, spoT-Δctd mutant Salmonella replicates poorly intracellularly and is attenuated in a murine model of acute salmonellosis. Collectively, these investigations indicate that (p)ppGpp synthesized by SpoT serves a unique function in the adaptation of Salmonella to the intracellular environment of host phagocytes that cannot be compensated by the presence of a functional RelA.IMPORTANCE Pathogenic bacteria experience nutritional challenges during colonization and infection of mammalian hosts. Binding of the alarmone nucleotide guanosine tetraphosphate (ppGpp) to RNA polymerase coordinates metabolic adaptations and virulence gene transcription, increasing the fitness of diverse Gram-positive and Gram-negative bacteria as well as that of actinomycetes. Gammaproteobacteria such as Salmonella synthesize ppGpp by the combined activities of the closely related RelA and SpoT synthetases. Due to its profound inhibitory effects on growth, ppGpp must be removed; in Salmonella, this process is catalyzed by the vital hydrolytic activity of the bifunctional SpoT protein. Because SpoT hydrolase activity is essential in cells expressing a functional RelA, we have a very limited understanding of unique roles these two synthetases may assume during interactions of bacterial pathogens with their hosts. We describe here a SpoT truncation mutant that lacks ppGpp synthetase activity and all C-terminal regulatory domains but retains excellent hydrolase activity. Our studies of this mutant reveal that SpoT uniquely senses the acidification of phagosomes, inducing virulence programs that increase Salmonella fitness in an acute model of infection. Our investigations indicate that the coexistence of RelA/SpoT homologues in a bacterial cell is driven by the need to mount a stringent response to a myriad of physiological and host-specific signatures.


Modulation of human eosinophil polymorphonuclear leukocyte migration and function.
Eosinophil migration toward a concentration gradient of a chemotactic factor is regulated at four levels. Diverse immunologic pathways generate stimuli with eosinophil chemotactic activity, including the complement products C5a and a fragment of C3a and the peptide products of mast cells and basophils activated by IgE-mediated reactions, such as eosinophil chemotactic factor of anaphylaxis (ECF-A) and other oligopeptides. The intrinsic preferential leukocyte activity of the chemotactic stimuli represents the second level of modulation, with ECF-A and other mast cell-derived peptides exhibiting the most selective action on eosinophils. The third level of control of eosinophil chemotaxis is composed of inactivators and inhibitors of chemotactic stimuli and is exemplified by degradation of C5a by anaphylatoxin inactivator or chemotactic factor inactivator and of ECF-A by carboxypeptidase-A or aminopeptidases. The activity of ECF-A is uniquely suppressed by equimolar quantities of its NH2- terminal tripeptide substituent, presumably by eosinophil membrane receptor competition. Factors comprising the fourth level of regulation, which alter eosinophil responsiveness to chemotactic stimuli, include the chemotactic factors themselves, through deactivation; nonchemotactic inhibitors such as the COOH-terminal tripeptide substituent of ECF-A, the neutrophil-immobilizing factor (NIF), the phagocytosis-enhancing factor Thr-Lys-Pro-Arg, and histamine at concentrations greater than 400 ng/ml; and nonchemotactic enhancing principles represented by ascorbate and by histamine at concentrations of 30 ng/ml or less. Local concentrations of eosinophils called to and immobilized at the site of a hypersenitivity reaction may express their regulatory functions by degrading the chemical mediators elaborated including histamine, slow-reacting substance of anaphylaxis (SRS-A), and platelet-activating factor (PAF) by way of their content of histaminase, arylsulfatase B, and phospholipase D, respectively. Immunologic pathways may thus provide the capability for early and specific host defense reactions with a later influx of eosinophils preventing irreversible local tissue alterations or distant organ effects.


Extracranial hypothermia during cardiac arrest and cardiopulmonary resuscitation is neuroprotective in vivo.
There is increasing evidence that ischemic brain injury is modulated by peripheral signaling. Peripheral organ ischemia can induce brain inflammation and injury. We therefore hypothesized that brain injury sustained after cardiac arrest (CA) is influenced by peripheral organ ischemia and that peripheral organ protection can reduce brain injury after CA and cardiopulmonary resuscitation (CPR). Male C57Bl/6 mice were subjected to CA/CPR. Brain temperature was maintained at 37.5°C ± 0.0°C in all animals. Body temperature was maintained at 35.1°C ± 0.1°C (normothermia) or 28.8°C ± 1.5°C (extracranial hypothermia [ExHy]) during CA. Body temperature after resuscitation was maintained at 35°C in all animals. Behavioral testing was performed at 1, 3, 5, and 7 days after CA/CPR. Either 3 or 7 days after CA/CPR, blood was analyzed for serum urea nitrogen, creatinine, alanine aminotransferase, aspartate aminotransferase, and interleukin-1β; mice were euthanized; and brains were sectioned. CA/CPR caused peripheral organ and brain injury. ExHy animals experienced transient reduction in brain temperature after resuscitation (2.1°C ± 0.5°C for 4 minutes). Surprisingly, ExHy did not change peripheral organ damage. In contrast, hippocampal injury was reduced at 3 days after CA/CPR in ExHy animals (22.4% ± 6.2% vs. 45.7% ± 9.1%, p=0.04, n=15/group). This study has two main findings. Hypothermia limited to CA does not reduce peripheral organ injury. This unexpected finding suggests that after brief ischemia, such as during CA/CPR, signaling or events after reperfusion may be more injurious than those during the ischemic period. Second, peripheral organ hypothermia during CA reduces hippocampal injury independent of peripheral organ protection. While it is possible that this protection is due to subtle differences in brain temperature during early reperfusion, we speculate that additional mechanisms may be involved. Our findings add to the growing understanding of brain-body cross-talk by suggesting that peripheral interventions can protect the brain even if peripheral organ injury is not altered.


Selective in vivo targeting of human liver tumors by optimized AAV3 vectors in a murine xenograft model.
Current challenges for recombinant adeno-associated virus (rAAV) vector-based cancer treatment include the low efficiency and the lack of specificity in vivo. rAAV serotype 3 (rAAV3) vectors have previously been shown to be ineffective in normal mouse tissues following systemic administration. In the present study, we report that rAAV3 vectors can efficiently target and transduce various human liver cancer cells in vivo. Elimination of specific surface-exposed serine and threonine residues on rAAV3 capsids results in further augmentation in the transduction efficiency of these vectors, without any change in the viral tropism and cellular receptor interactions. In addition, we have identified a potential chemotherapy drug, shikonin, as a multifunctional compound to inhibit liver tumor growth as well as to significantly enhance the efficacy of rAAV vector-based gene therapy in vivo. Furthermore, we also document that suppression of tumorigenesis in a human liver cancer xenograft model can be achieved through systemic administration of the optimized rAAV3 vectors carrying a therapeutic gene, and shikonin at a dose that does not lead to liver damage. Our research provides a novel means to achieve not only targeted delivery but also the potential for gene therapy of human liver cancer.


Effects of protein-calorie malnutrition on transplacental kinetics of aminoisobutyric acid in rats.
In order to find out if inefficient transport of amino acids contributes to a decrease in fetal weight during maternal malnutrition, we injected [14C]- and [3H]-labelled aminoisobutyric acid (AIB), respectively, in the mother and its fetuses and determined its transplacental kinetics on day 20 of gestation in rats fed a 21 per cent (control) or a low (5 per cent) protein diet. Rats fed a low protein diet consumed significantly less food than did the rats fed a control diet and thus suffered from protein-calorie malnutrition. A low protein diet led to a significant (P less than 0.05) decrease in maternal and fetal volume of distribution of AIB, a decrease in the clearance of AIB from the mother to the fetus and an increase in the time required for the fetal plasma AIB concentration following maternal injection to exceed the maternal plasma AIB concentration. The clearance of AIB from the fetus into the mother or to outside (e.g. amniotic fluid) was not altered by protein deficiency. It is concluded that a decrease in the efficiency of the placenta to deliver amino acids to the fetus may be a contributing factor in fetal growth retardation during maternal protein malnutrition.


Terahertz time-domain spectroscopy of human blood.
In the continuing development of terahertz technology to enable the determination of tissue pathologies in real-time during surgical procedures, it is important to distinguish the measured terahertz signal from biomaterials and fluids, such as blood, which may mask the signal from tissues of interest. In this paper, we present the frequency-dependent absorption coefficients, refractive indices, and Debye relaxation times of whole blood, red blood cells, plasma, and a thrombus.


Cooccupancy of the outer vestibule of voltage-gated sodium channels by micro-conotoxin KIIIA and saxitoxin or tetrodotoxin.
The guanidinium alkaloids tetrodotoxin (TTX) and saxitoxin (STX) are classic ligands of voltage-gated sodium channels (VGSCs). Like TTX and STX, micro-conotoxin peptides are pore blockers but with greater VGSC subtype selectivity. micro-Conotoxin KIIIA blocks the neuronal subtype Na(V)1.2 with nanomolar affinity and we recently discovered that KIIIA and its mutant with one fewer positive charge, KIIIA[K7A], could act synergistically with TTX in a ternary peptide x TTX x Na(V) complex. In the complex, the peptide appeared to trap TTX in its normal binding site such that TTX could not readily dissociate from the channel until the peptide had done so; in turn, the presence of TTX accelerated the rate at which peptide dissociated from the channel. In the present study we examined the inhibition of Na(V)1.2, exogenously expressed in Xenopus oocytes, by STX (a divalent cation) and its sulfated congener GTX2/3 (with a net +1 charge). Each could form a ternary complex with KIIIA and Na(V)1.2, as previously found with TTX (a monovalent cation), but only when STX or GTX2/3 was added before KIIIA. The KIIIA x alkaloid x Na(V) complex was considerably less stable with STX than with either GTX2/3 or TTX. In contrast, ternary KIIIA[K7A] x alkaloid x Na(V) complexes could be formed with either order of ligand addition and were about equally stable with STX, GTX2/3, or TTX. The most parsimonious interpretation of the overall results is that the alkaloid and peptide are closely apposed in the ternary complex. The demonstration that two interacting ligands ("syntoxins") occupy adjacent sites raises the possibility of evolving a much more sophisticated neuropharmacology of VGSCs.


Family meals and eating practices among mothers in Santos, Brazil: A population-based study.
This study investigates family meals among mothers and explores associations between eating with family and sociodemographic characteristics, body mass index, and eating practices. A population-based cross-sectional study, using complex cluster-sampling, was conducted in the city of Santos, Brazil with 439 mothers. Frequency of family meals was assessed by asking if mothers did or did not usually have a) breakfast, b) lunch, and c) dinner with family. Linear regression analyses were conducted for the number of meals eaten with family per day and each of the potential explanatory variables, adjusting for the mother's age. Poisson regression with robust variance was used to analyze each factor associated with eating with family as classified categorically: a) sharing meals with family, b) not eating any meals with family. Only 16.4% (n = 72) of participants did not eat any meals with family. From the 83.6% (n = 367) of mothers that had at least one family meal per day, 69.70% (n = 306) ate dinner with their families. Mothers aged ≥40 years reported significantly fewer meals eaten with family compared to mothers aged 30-39 years (β: -0.26, p = 0.04). Having family meals was 54% more prevalent among mothers with ≥12 years of education (PR for no meals eaten with family: 0.54, 95% CI: 0.30; 0.96, p = 0.03), when compared to mothers with less than nine years of education. Eating no meals with family was 85% more prevalent among mothers who reported that eating was one of the biggest pleasures in their lives (PR: 1.85, 95% CI: 1.21; 2.82, p = 0.004). We suggest the need for further research investigating the effects of family meals on mothers' health through nutritional and phenomenological approaches.


Change in intraocular pressure measurement 2 years after myopic laser-assisted subepithelial keratectomy.
To evaluate intraocular pressure (IOP) measurements 24 months after laser-assisted subepithelial keratectomy (LASEK) with 3 tonometry methods-Goldmann applanation tonometry (GAT), Pascal dynamic contour tonometry (DCT), and applanation resonance tonometry (ART)-and to compare them with measurements taken preoperatively and 3 and 6 months postoperatively. Department of Ophthalmology, Umeå University, Umeå, Sweden. Prospective randomized single-center. One randomized eye per person from myopic healthy individuals who had LASEK was included. Visual acuity, central corneal thickness (CCT), corneal curvature, and IOP were measured at all time points. Six IOP measurements were performed per method, with a 5-minute lapse between methods. The study evaluated 38 eyes. Two years after LASEK, dynamic ART was the only method that did not measure significantly lower IOP than before LASEK (mean -0.2 mm Hg ± 1.6 [SD]). The greatest difference was with GAT (mean -1.9 ± 1.7 mm Hg) followed by static ART (mean -1.2 ± 1.6 mm Hg) and then DCT (mean -0.9 ± 1.3 mm Hg). All methods measured significantly lower IOP at 3 months and 6 months. Dynamic ART, which analyses IOP measurement during indentation, showed no significant difference in measured IOP after 24 months. The convex tip of the ART device and the continuous sampling of data during corneal indentation seem to be factors in why dynamic ART was less affected by corneal properties and therefore more suitable for IOP measurement in LASEK-treated eyes, although this must be confirmed in larger trials.


Can blood pressure be lowered safely in older adults with lacunar stroke? The Secondary Prevention of Small Subcortical Strokes study experience.
To determine safety and tolerability of lowering blood pressure in older adults with lacunar stroke. Cohort study. The Secondary Prevention of Small Subcortical Strokes (SPS3) Trial, which compared the efficacy of two systolic blood pressure (SBP) targets (<130 mmHg and 130-149 mmHg) for secondary stroke prevention. Of 3,020 SPS3 participants, 494 aged 75 and older at baseline were used in these analyses. Rates of side effects related to lowering SBP and clinical outcomes, including stroke recurrence and vascular death, were examined. Older participants achieved SBP levels similar to those of younger participants (mean SBP of 125 mmHg and 137 mmHg in lower and higher SBP target groups, respectively). At least once during the approximately 3.5 years of follow-up, 21% reported dizziness, and 15% reported lightheadedness when standing; the only significant difference between the younger and older groups was unsteadiness when standing (23% vs 32% respectively, P < .001). There was no difference according to treatment group. In younger adults, recurrent stroke was less likely in the lower than the higher SBP group (hazard ratio (HR) = 0.77, 95% confidence interval (CI) = 0.59-1.01) but not in older participants (HR = 1.01, 95% CI = 0.59-1.73), although the interaction was not significant (P = .39). The lower SBP target was associated with a significant reduction in vascular death in older participants (HR = 0.42, 95% CI = 0.18-0.98), with a significant interaction between age and SBP group (P = .049). Except for unsteadiness when standing, there was no difference according to age in individuals with lacunar stroke with respect to side effects potentially related to lowering blood pressure. Although the lower SBP target was not associated with lower likelihood of recurrent stroke, these exploratory analyses suggested a possible benefit related to vascular death.


Rapid inactivation of chloroplastic ascorbate peroxidase is responsible for oxidative modification to Rubisco in tomato (Lycopersicon esculentum) under cadmium stress.
To investigate the sensitive site of antioxidant systems in chloroplast under cadmium stress and its consequence on reactive oxygen species production and action, the sub-organellar localization of chloroplast superoxide dismutases (SOD, EC 22.214.171.124) and ascorbic peroxidase (APX, EC 22.214.171.124) isoenzymes and changes of enzymes activities under cadmium stress were investigated in tomato seedlings. Two APX isoforms, one thylakoid-bound and one stromal, were detected. Cd at 50 microM induced a moderate increase of SOD activities but a rapid inactivation of both APX isoenzymes. APX inactivation was mainly related to the decrease of ascorbate concentration, as supported by in vitro treatment of exogenous ascorbate and APX kinetic properties under Cd stress. H2O2 accumulation in chloroplast, as a consequence of APX inactivation, was associated with a 60% loss of Rubisco (EC 18.104.22.168) activity, which could be partially accounted for by a 10% loss of Rubisco content. Protein oxidation assay found that the Rubisco large subunit was the most prominent carbonylated protein; the level of carbonylated Rubisco large subunit increased fivefold after Cd exposure. Thiol groups in the Rubisco large subunit were oxidized, as indicated by non-reducing electrophoresis. Treating crude extract with H2O2 resulted in a similar pattern of protein oxidation and thiols oxidation with that observed in Cd-treated plants. Our study indicates that APXs in the chloroplast is a highly sensitive site of antioxidant systems under Cd stress, and the inactivation of APX could be mainly responsible for oxidative modification to Rubisco and subsequent decrease in its activity.


Highly Selective Olefin-Assisted PdII -Catalyzed Oxidative Alkynylation of Enallenes.
An olefin-assisted, palladium-catalyzed oxidative alkynylation of enallenes for regio- and stereoselective synthesis of substituted trienynes has been developed. The reaction shows a broad substrate scope and good tolerance for various functional groups on the allene moiety, including carboxylic acid esters, free hydroxyls, imides, and alkyl groups. Also, a wide range of terminal alkynes with electron-donating and electron-withdrawing aryls, heteroaryls, alkyls, trimethylsilyl, and free hydroxyl groups are tolerated.


[Relation of the maximum calf circumference with albumin and body mass index in elderly cases of chronic hospitalization].
To examine the relation of the calf circumference at the level where the size of the triceps surae appeared largest (maximum calf circumference MCC) with albumin (Alb) and body mass index (BMI) in elderly cases of chronic hospitalization. Subjects were 57 elderly cases of chronic hospitalization (26 men and 31 women aged 80.4+/-9.9 years [mean+/-SD]) who varied in their eating styles and degrees of independence in eating, but still could ingest orally. We compared the subjects' weight, height, BMI, MCC, and Alb between the both sexes with t-test. The relationship between the MCC and Functional Independence Measure (FIM) motor score, FIM cognitive score, and FIM total score were analyzed with Spearman's rank correlation coefficient, and Alb and BMI with Pearson's correlation analysis. Additionally, the partial correlation coefficient between the MCC and Alb adjusted for sex, age, and FIM total score was calculated. Sex-related difference was observed in height and weight (p<0.01), but not in BMI, MCC, and Alb. The MCC did not significantly correlate with motor, cognitive, and total FIMs, with correlation coefficients of r=0.20, r=0.19, and r=0.23, respectively. The MCC significantly correlated with Alb (r=0.52, p<0.01) and BMI (r=0.90, p<0.01). The regression equation to estimate BMI from the MCC was BMI (kg/m(2))=MCC x 0.81-2.44, with the coefficient of determination being 0.81. The partial correlation coefficient between the MCC and Alb was r=0.40 (p<0.01). This study suggests that the MCC may be a useful evaluation tool for BMI in elderly people with chronic conditions. The value may also reflect the systemic nutritional status, and deserves further study.


Ubiquitin-activating enzyme E1 inhibitor PYR41 attenuates angiotensin II-induced activation of dendritic cells via the IκBa/NF-κB and MKP1/ERK/STAT1 pathways.
The activation of dendritic cells (DCs) is necessary to initiate immune responses. Angiotensin II (Ang II) can enhance the maturation and activation of DCs, but the mechanisms are still unclear. Ubiquitin-activating enzyme (E1/Uba1) is the common first step in ubiquitylation, which decides whether or not the modified protein is ultimately degraded by the proteasome. This study aimed to investigate the role of E1 in Ang II-induced activation of DCs and the underlying mechanisms. First, we showed that Ang II stimulation significantly up-regulated E1 expression in DCs. Moreover, Ang II treatment markedly induced phenotypic maturation, the secretion of cytokines and the immunostimulatory capacity of DCs. In contrast, inhibition of E1 by a small molecule inhibitor, 4 [4-(5-nitro-furan-2-ylmethylene)-3, 5-dioxo-pyrazolidin-1-yl]-benzoic acid ethyl ester (PYR41), markedly attenuated these effects. Mechanistically, PYR41 treatment markedly decreased K63-linked ubiquitination of tumour necrosis factor receptor-associated factor 6 and nuclear factor-κB essential modulator, inhibited proteasomal degradation of nuclear factor-κB inhibitor α and mitogen-activated protein kinase phosphatase 1 thereby resulting in activation of nuclear factor-κB, extracellular signal-regulated kinase 1/2 and signal transducer and activator of transcription 1 signalling pathways in DCs induced by Ang II. Taken together, our results demonstrate a novel role of E1 in Ang II-induced activation of DCs, and inhibition of E1 activity might be a potential therapeutic target for DC-mediated autoimmune diseases.


The expression and secretion of vimentin in the progression of non-alcoholic steatohepatitis.
The pathogenesis of non-alcoholic steatohepatitis (NASH) is not fully understood. In the present study, both in vitro and in vivo vimentin expression and secretion in NASH were investigated. The exposure of palmitate and lipopolysaccharide (LPS) to HepG2 cells enhanced caspase-3 activity and vimentin expression, respectively. The combined effects of both treatments on vimentin expression and caspase-3 activation appeared to be synergic. In contrast, blockade of caspase-3 activity by zVADfmk resulted in a significant reduction of cleaved vimentin and secreted vimentin into the culture supernatant. Similarly, lipid accumulation and inflammation occurred in mice fed a methionine-choline-deficient diet; thus, vimentin expression and serum cleaved vimentin levels were increased. However, vimentin was not significantly upregulated, and no cleavage occurred in mice fed a high-fat diet. It was conclusively determined that lipid accumulation in hepatocytes induces apoptosis through a caspase-3 dependent pathway; whereas, LPS stimulates vimentin expression, leading to its cleavage and secretion. Increased vimentin fragment levels indicated the existence of substantial hepatocellular death via an apoptotic mechanism.


Brief exposure to intense turbulence induces a sustained life-history shift in echinoids.
In coastal ecosystems, attributes of fluid motion can prompt animal larvae to rise or sink in the water column and to select microhabitats within which they attach and commit to a benthic existence. In echinoid (sea urchin and sand dollar) larvae living along wave-exposed shorelines, intense turbulence characteristic of surf zones can cause individuals to undergo an abrupt life-history shift characterized by precocious entry into competence - the stage at which larvae will settle and complete metamorphosis in response to local cues. However, the mechanistic details of this turbulence-triggered onset of competence remain poorly defined. Here, we evaluate in a series of laboratory experiments the time course of this turbulence effect, both the rapidity with which it initiates and whether it perdures. We found that larvae become competent with turbulence exposures as brief as 30 s, with longer exposures inducing a greater proportion of larvae to become competent. Intriguingly, larvae can remember such exposures for a protracted period (at least 24 h), a pattern reminiscent of long-term potentiation. Turbulence also induces short-term behavioral responses that last less than 30 min, including cessation of swimming, that facilitate sinking and thus contact of echinoid larvae with the substratum. Together, these results yield a novel perspective on how larvae find their way to suitable adult habitat at the critical settlement transition, and also open new experimental opportunities to elucidate the mechanisms by which planktonic animals respond to fluid motion.


Identification of substrain-specific mutations by massively parallel whole-genome resequencing of Synechocystis sp. PCC 6803.
The cyanobacterium, Synechocystis sp. PCC 6803, was the first photosynthetic organism whose genome sequence was determined in 1996 (Kazusa strain). It thus plays an important role in basic research on the mechanism, evolution, and molecular genetics of the photosynthetic machinery. There are many substrains or laboratory strains derived from the original Berkeley strain including glucose-tolerant (GT) strains. To establish reliable genomic sequence data of this cyanobacterium, we performed resequencing of the genomes of three substrains (GT-I, PCC-P, and PCC-N) and compared the data obtained with those of the original Kazusa strain stored in the public database. We found that each substrain has sequence differences some of which are likely to reflect specific mutations that may contribute to its altered phenotype. Our resequence data of the PCC substrains along with the proposed corrections/refinements of the sequence data for the Kazusa strain and its derivatives are expected to contribute to investigations of the evolutionary events in the photosynthetic and related systems that have occurred in Synechocystis as well as in other cyanobacteria.


Combined multivariate data analysis of high-performance thin-layer chromatography fingerprints and direct analysis in real time mass spectra for profiling of natural products like propolis.
Sophisticated statistical tools are required to extract the full analytical power from high-performance thin-layer chromatography (HPTLC). Especially, the combination of HPTLC fingerprints (image) with chemometrics is rarely used so far. Also, the newly developed, instantaneous direct analysis in real time mass spectrometry (DART-MS) method is perspective for sample characterization and differentiation by multivariate data analysis. This is a first novel study on the differentiation of natural products using a combination of fast fingerprint techniques, like HPTLC and DART-MS, for multivariate data analysis. The results obtained by the chemometric evaluation of HPTLC and DART-MS data provided complementary information. The complexity, expense, and analysis time were significantly reduced due to the use of statistical tools for evaluation of fingerprints. The approach allowed categorizing 91 propolis samples from Germany and other locations based on their phenolic compound profile. A high level of confidence was obtained when combining orthogonal approaches (HPTLC and DART-MS) for ultrafast sample characterization. HPTLC with selective post-chromatographic derivatization provided information on polarity, functional groups and spectral properties of marker compounds, while information on possible elemental formulae of principal components (phenolic markers) was obtained by DART-MS.


[Functional state of peripheral blood neutrophils in children with rheumatism and borderline conditions].
The authors discuss the effect of exotoxin of beta-hemolytic streptococcus (streptolysin-O) on the chemotactic activity of the peripheral blood leukocytes. 107 children with various forms of rheumatic fever and acute diseases of Streptococcal etiology and 18 healthy children were examined. The absence of the phenomenon of a specific chemotactic deactivation of polymorphonuclear leukocytes in response to streptolysin-O in healthy persons and in those suffering from rheumatic fever has been revealed. Sharply marked sensitization of polymorphonuclear leukocytes to streptolysin-O in patients with rheumatism increasing in the course of the disease and remaining in the inactive phase has been noted. The functional activity of polymorphonuclear leukocytes is characterized by chemotactic deactivation in response to streptolysin O that probably provides a favorable outcome of the disease.


Linking lysosomal biomarker and population responses in a field population of Aporrectodea caliginosa (Oligochaeta) exposed to the fungicide copper oxychloride.
The effects of the fungicide copper oxychloride on a field population of the earthworm Aporrectodea caliginosa were investigated. Changes were monitored in earthworm numbers and biomass, copper concentrations in soil and earthworms, and neutral red retention times (NRRTs) of coelomocytes. The aim was to establish whether the cellular responses could be linked to changes at the population level. Results indicated that there was a significant (P<0.05) impact on worm density and biomass on the treated plots 6 months after spraying with the fungicide stopped. NRRTs of treated earthworms were significantly lower after one spraying when earthworms had elevated concentrations of copper in their body tissues. Six months after spraying stopped, copper levels in both soil and earthworm tissues declined significantly, but the earthworm numbers and biomass were still significantly lower in the treated plots. This indicated that the effects of the copper oxychloride exposure manifested at a much later stage at the population level, than on NRRT. We conclude that the decrease in NRRTs, which preceded the decrease in both earthworm biomass and numbers, may have predictive value as a biomarker of impending population changes.


[Monolayer culture of bone marrow and splenic fibroblasts in lymphoproliferative diseases].
The authors describe the results of monolayer cultivation bone marrow and spleen fibroblasts in patients with lymphoproliferative diseases. The efficacy of colony-formation of fibroblasts (ECFf) was studied in 10 patients with chronic lympholeukemia, 4 with leukemic reticuloendotheliosis, in 5 with lymphocytoma of the spleen, and in 3 with lymphosarcoma of the spleen. The control group comprised 7 normal subjects. Comparison of the ECFf on the spleen and bone marrow during lymphocytomas demonstrated the ECFf in the bone marrow to be low (0.24 +/- 0.16) in contrast to the high and normal ECFf in chronic lympholeukemia and leukemic reticuloendotheliosis. The ECFf in the spleen of patients with lymphocytomas was high (4.20 +/- 0.90), i. e. higher than in the control group (1.1 +/- 0.31). In lymphosarcoma of the spleen, the growth of fibroblasts was low in both the bone marrow and the spleen (0.15 +/- 0.13 and 0.10 +/- 0.01). These data suggest that the growth of stromal elements in lymphatic tumors is determined by the characteristic features of tumor cells.


Correlation of contrast-enhanced MR images with the histopathology of minimally invasive thermal and cryoablation cancer treatments in normal dog prostates.
Magnetic Resonance Imaging (MRI) is a promising tool for visualizing the delivery of minimally invasive cancer treatments such as high intensity ultrasound (HUS) and cryoablation. We use an acute dog prostate model to correlate lesion histopathology with contrast-enhanced (CE) T1 weighted MR images, to aid the radiologists in real time interpretation of in vivo lesion boundaries and pre-existing lesions. Following thermal or cryo treatments, prostate glands are removed, sliced, stained with the vital dye triphenyl tetrazolium chloride, photographed, fixed and processed in oversized blocks for routine microscopy. Slides are scanned by Trestle Corporation at .32 microns/pixel resolution, the various lesions traced using annotation software, and digital images compared to CE MR images. Histologically, HUS results in discrete lesions characterized by a "heat-fixed" zone, in which glands subjected to the highest temperatures are minimally altered, surrounded by a rim or "transition zone" composed of severely fragmented, necrotic glands, interstitial edema and vascular congestion. The "heat-fixed" zone is non-enhancing on CE MRI while the "transition zone" appears as a bright, enhancing rim. Likewise, the CE MR images for cryo lesions appear similar to thermally induced lesions, yet the histopathology is significantly different. Glands subjected to prolonged freezing appear totally disrupted, coagulated and hemorrhagic, while less intensely frozen glands along the lesion edge are partially fragmented and contain apoptotic cells. In conclusion, thermal and cryo-induced lesions, as well as certain pre-existing lesions (cystic hyperplasia - non-enhancing, chronic prostatitis - enhancing) have particular MRI profiles, useful for treatment and diagnostic purposes.


QCM sensing of miR-21 by formation of microRNA-DNA hybrid duplexes and intercalation on surface-functionalized pyrene.
MicroRNAs (miRNAs) are small non-coding RNA molecules that serve as important biomarkers for a variety of diseases such as cancer and vascular disease. However, sensitive and accurate detection of miR-21 is very challenging in that up-regulation of miR-21 is highly associated with several types of malignant tumors. Here, quartz crystal microbalance (QCM) biosensors were developed for sensitive and specific detection of miR-21 through formation of miR-21-DNA hybrid duplexes and non-specific intercalation of surface-modified pyrene molecules. High selectivity for miR-21 over other miRNAs came from the specific hybridization between miR-21 and gold nanoparticle (AuNP)-conjugated complementary oligonucleotides of miR-21. High sensitivity was obtained through formation of intercalated complexes on the surface with subsequent gold staining signal amplification. Under optimum condition using this strategic approach, our novel QCM biosensors could detect miR-21 concentration as low as 3.6 pM in the entire linear range from 2.5 pM to 2.5 μM with a correlation coefficient of 0.989. In addition, these sensors did not work at all for other miRNAs based on their high selectivity. miR-21 in human brain total RNA and total RNA extracted from A549 cell line could also be successfully detected. Therefore, miRNA detection technology using QCM biosensors and their detection mechanisms have potential as alternatives in biological studies and clinical diagnosis.


Full-length transcriptome characterization and comparative analysis of Gleditsia sinensis.
As an economically important tree, Gleditsia sinensis Lam. is widely planted. A lack of background genetic information on G. sinensis hinders molecular breeding. Based on PacBio single-molecule real-time (SMRT) sequencing and analysis of G. sinensis, a total of 95,183 non-redundant transcript sequences were obtained, of which 93,668 contained complete open reading frames (ORFs), 2,858 were long non-coding RNAs (LncRNAs) and 18,855 alternative splicing (AS) events were identified. Genes orthologous to different Gleditsia species pairs were identified, stress-related genes had been positively selected during the evolution. AGA, AGG, and CCA were identified as the universal optimal codon in the genus of Gleditsia. EIF5A was selected as a suitable fluorescent quantitative reference gene. 315 Cytochrome P450 monooxygenases (CYP450s) and 147 uridine diphosphate (UDP)-glycosyltransferases (UGTs) were recognized through the PacBio SMRT transcriptome. Randomized selection of GsIAA14 for cloning verified the reliability of the PacBio SMRT transcriptome assembly sequence. In conclusion, the research data lay the foundation for further analysis of the evolutionary mechanism and molecular breeding of Gleditsia.


Assessment of health management information system for monitoring of maternal health in Jaleswar Block of Balasore District, Odisha, India.
In 2005, the Government of India implemented the National Rural Health Mission for reduction of maternal mortality. One of the major impediments in improving maternal health since then has been a poor management of the Health Management Information System (HMIS) at grass-roots level which could integrate data collection, processing, reporting, and use of information for necessary improvement of health services. The paper identifies the challenges in generating information for HMIS and its utilization for improvement of maternal health program in the tribal-dominated Jaleswar block in Odisha, India. It also aims to understand the nature and orientation of the HMIS data generated by the government for the year 2013-2014. The study is a cross-sectional type which used observation and interview methods. Primary data were gathered from health professionals to understand the challenges in generating information for HMIS and its utilization. Next, to understand the nature and orientation of HMIS, data pertaining to tribal block were analyzed. The findings show that there are challenges in generation of quality data, capacity building of workforce, and monitoring of vulnerable tribal population. The discrepancies between HMIS data and field reality display the gap in formulation of policy and its implementation. The study unearths the existing politics of knowledge generation. This shows highly standardized procedures and information gathering by use of dominant biomedical concepts of maternal health with limited inclusion of local birthing conceptions and needs of vulnerable tribal pregnant women.


Three-Dimensional Organization of Chicken Genome Provides Insights into Genetic Adaptation to Extreme Environments.
The high-throughput chromosome conformation capture (Hi-C) technique is widely used to study the functional roles of the three-dimensional (3D) architecture of genomes. However, the knowledge of the 3D genome structure and its dynamics during extreme environmental adaptations remains poor. Here, we characterized 3D genome architectures using the Hi-C technique for chicken liver cells. Upon comparing Lindian chicken (LDC) liver cells with Wenchang chicken (WCC) liver cells, we discovered that environmental adaptation contributed to the switching of A/B compartments, the reorganization of topologically associated domains (TADs), and TAD boundaries in both liver cells. In addition, the analysis of the switching of A/B compartments revealed that the switched compartmental genes (SCGs) were strongly associated with extreme environment adaption-related pathways, including tight junction, notch signaling pathway, vascular smooth muscle contraction, and the RIG-I-like receptor signaling pathway. The findings of this study advanced our understanding of the evolutionary role of chicken 3D genome architecture and its significance in genome activity and transcriptional regulation.


[The effects of pregabaline on postoperative pain and opioid consumption used perioperatively in patients undergoing modified radical mastectomy].
In this study, effects of pregabaline on postoperative pain and opioid consumption used perioperatively in patients undergoing modified radical mastectomy(MRM) were investigated. Sixty ASA 1-2 patients scheduled for MRM were included. Patients were randomly divided into two and 30 patients were allocated into each group. Group Pregabaline was given pregabaline 150 mg 1 hr before operation and Group Placebo empty capsule. In both groups, anesthesia induction was obtained by penthotal, fentanyl and rocuronium and maintainence by sevoflurane, N2O and O2. Twelve hr after operation, Group Pregabaline was administered pregabaline 75 mg while Placebo group received empty capsule again. All patients received lornoxicam 8 mg iv 1 hr before end of surgery and ondansetrone 4 mg 30 min before. Patient-controlled analgesia device prepared with morphine was connected to both groups for postoperative analgesia. Postoperative VAS pain scores, hemodynamic parameters, morphine consumption, side-effects like nausea-vomiting, sedation and dizziness were followed and recorded. Demographic parameters were similar. VAS scores were significantly lower in Group Pregabaline at 1, 30 min, 1,4,8 and 12 hr (p<0.05). There was no significant difference in postoperative morphine consumption and need for additional dose, although they were higher in Placebo group. The patients in Placebo group had higher bothersome scores for side-effects. Hemodynamic parameters and other side-effects were similar. In our study, we showed that pregabalin administered perioperatively increased postoperative analgesic efficacy in MRM operations without making significant side effect, but did not change opioid consumption. We think that further studies about this topic must be held with different dose and patient groups.


A Comparative Evaluation of Lateral Condylar Guidance by Clinical and Radiographic Methods - Hanau's Formula Revisited.
Various clinical methods for recording the condylar guidance (CG) are the intraoral check bite method, graphic tracings, and functional recordings. Accuracy of graphic tracings is affected by patient-related factors such as neuromuscular control of the individual, stability of record bases as well as stability of recording media. The current recommended average settings using Hanau's formula questionable, and thus reassessment is needed. The purpose of this study is to use radiographic technique to determine the lateral CG and compare these values with those obtained using Hanau's formula and to evaluate whether there are differences between the right and left paths of the condyles. Twenty completely edentulous patients were selected. Articulator was modified with sectioned protractor for obtaining per degree interval calibration. With the protrusive records, the horizontal CG (H) was adjusted and the Bennett's angle (LCG-C) was calculated using the formula. With the tracing device in the mouth, sub-mento vertex projection radiographs were obtained. Each radiograph was traced and superimposed for Bennett angle determination (LCG-M). The median (interquartile range [IQR]) of right and left LCG-C were 15.45 (0.8) and 15.50 (0.7), respectively. The median (IQR) of right and left LCG-M were 37.00 (6.0) and 36.50 (6.8), respectively. A statistically significant difference exists between LCG-C and LCG-M. Both LCG-C and-M values exhibited no variations on both sides. Radiographic technique yielded an amplified LCG when compared to the value obtained by Hanau's formula.


Antimicrobial properties of amyloid peptides.
More than two dozen clinical syndromes known as amyloid diseases are characterized by the buildup of extended insoluble fibrillar deposits in tissues. These amorphous Congo red staining deposits known as amyloids exhibit a characteristic green birefringence and cross-β structure. Substantial evidence implicates oligomeric intermediates of amyloids as toxic species in the pathogenesis of these chronic disease states. A growing body of data has suggested that these toxic species form ion channels in cellular membranes causing disruption of calcium homeostasis, membrane depolarization, energy drainage, and in some cases apoptosis. Amyloid peptide channels exhibit a number of common biological properties including the universal U-shape β-strand-turn-β-strand structure, irreversible and spontaneous insertion into membranes, production of large heterogeneous single-channel conductances, relatively poor ion selectivity, inhibition by Congo red, and channel blockade by zinc. Recent evidence has suggested that increased amounts of amyloids not only are toxic to its host target cells but also possess antimicrobial activity. Furthermore, at least one human antimicrobial peptide, protegrin-1, which kills microbes by a channel-forming mechanism, has been shown to possess the ability to form extended amyloid fibrils very similar to those of classic disease-forming amyloids. In this paper, we will review the reported antimicrobial properties of amyloids and the implications of these discoveries for our understanding of amyloid structure and function.


Clinical Conditions and Prescription Drug Utilization among Early Medical Marijuana Registrants in Florida.
Initial legalization of medical marijuana (MM) in Florida required providers to submit initial and follow-up treatment plan forms to the University of Florida to support research on MM safety and efficacy. This study retrospectively analyzed all treatment plan forms submitted between program inception (August 2016) through July 2017 and describes early Florida MM registrants by clinical conditions and prescription drug utilization. Among 7,548 unique treatment plans, the initial visit was characterized by registrants who were mostly white (83.7%), 52.3 (SD 16.4) years of age on average, and who were assessed by the provider as at least moderately ill (79.6%). Musculoskeletal and spasticity-related conditions (44.8%), chronic pain (41.9%), and mental health disorders (17.0%) were the most frequent medical complaints for seeking MM treatment with more than one condition per patient possible. One in four (25.9%) patients reported use of prescription opioids and over one-fifth of patients frequently utilized at least one psychotropic medication as well as cardiovascular agents. There were 2,075 unique follow-up plans available which were mostly characterized by clinical improvement and reported reductions in utilization of some drug classes. Further research is needed to guide clinicians on the risks and benefits of MM used concomitantly with prescription drugs.


PedsQL eosinophilic esophagitis module: feasibility, reliability, and validity.
Eosinophilic esophagitis (EoE) is a chronic esophageal inflammatory condition with a paucity of information on health-related quality of life (HRQOL). The objective of the study was to report on the measurement properties of the PedsQL EoE Module. The PedsQL EoE Module was completed in a multisite study by 196 pediatric patients with EoE and 262 parents of patients with EoE. The PedsQL EoE Module scales evidenced excellent feasibility (0.6%-3.1% missing), excellent group comparison reliability across total scale scores (patient α 0.93; parent proxy α 0.94), good reliability for the 7 individual scales (patient α 0.75-0.87; parent proxy α 0.81-0.92), excellent test-retest reliability (patient intraclass correlation coefficient 0.88; parent intraclass correlation coefficient 0.82), demonstrated no floor effects and low ceiling effects, and demonstrated a high percentage of scaling success for most scales. Intercorrelations with the PedsQL Generic Core Scales were in the medium (0.30) to large (0.50) range. PedsQL EoE Module scores were worse among patients with active histologic disease (≥ 5 eos/hpf) compared with those in remission (patient self-report: 63.3 vs 69.9 [P < 0.05]; parent proxy report: 65.1 vs 72.3 [P < 0.01]), and those treated with dietary restrictions compared with those with no restrictions (patient self-report: 61.6 vs 74.3 [P < 0.01]; parent proxy report: 65.5 vs 74.7 [P < 0.01]). The results demonstrate excellent measurement properties of the PedsQL EoE Module. Patients with active histologic disease and those treated with dietary restrictions demonstrated worse PedsQL scores. The PedsQL EoE Module may be used in the evaluation of pediatric EoE disease-specific HRQOL in clinical research and practice.


Biphasic binding kinetics between FepA and its ligands.
The Escherichia coli FepA protein is an energy- and TonB-dependent, ligand-binding porin that functions as a receptor for the siderophore ferric enterobactin and colicins B and D. We characterized the kinetic and thermodynamic parameters associated with the initial, energy-independent steps in ligand binding to FepA. In vivo experiments produced Kd values of 24, 185, and 560 nM for ferric enterobactin, colicin B, and colicin D, respectively. The siderophore and colicin B bound to FepA with a 1:1 stoichiometry, but colicin D bound to a maximum level that was 3-fold lower. Preincubation with ferric enterobactin prevented colicin B binding, and preincubation with colicin B prevented ferric enterobactin binding. Colicin B release from FepA was unexpectedly slow in vivo, about 10-fold slower than ferric enterobactin release. This slow dissociation of the colicin B.FepA complex facilitated the affinity purification of FepA and FepA mutants with colicin B-Sepharose. Analysis of a fluorescent FepA derivative showed that ferric enterobactin and colicin B adsorbed with biphasic kinetics, suggesting that both ligands bind in at least two distinct steps, an initial rapid stage and a subsequent slower step, that presumably establishes a transport-competent complex.


Using the Interpersonal Skills tool to assess interpersonal skills of internationally educated nurses.
To assess interpersonal skills of internationally educated nurses (IEN) while interacting with standardized patients. Participants included 52 IEN at two community hospitals in the southwestern region of the USA. Standardized patients were used to create patient-nurse encounter. Seventeen items in four domains ("skills in interviewing and collecting information"; "skills in counseling and delivering information"; "rapport"; and "personal manner") in an Interpersonal Skills (IPS) instrument were measured by a Likert scale 1-4 with 4 indicating the best performance. The average composite score per domain and scores of the 17 items were compared across the domains. On 10 of the 17 items, the nurses received scores under 3. Counseling with an average score of 2.10 and closure with an average score of 2.44 in domain 2, small talk with an average score of 2.06 in domain 3, and physical exam with average score of 2.21 in domain 4 were below 2.5. The average composite score of domain 1 was 3.54, significantly higher than those of domains 2-4 (2.77, 2.81, and 2.71, respectively). Age was moderately related to the average score per domain with every 10 year increase in age resulting in a 0.1 increase in the average score. Sex and country of origin showed mixed results. The interpersonal skills of IEN in three of the four domains need improvement. Well-designed educational programs may facilitate the improvement, especially in areas of small talk, counseling, closure, and physical exam. Future research should examine relationships between the IPS and demographics factors.


The de novo synthesis of GDP-fucose is essential for flagellar adhesion and cell growth in Trypanosoma brucei.
The protozoan parasite Trypanosoma brucei causes human African sleeping sickness in sub-Saharan Africa. The parasite makes several essential glycoproteins, which has led to the investigation of the sugar nucleotides and glycosyltransferases required to synthesize these structures. Fucose is a common sugar in glycoconjugates from many organisms; however, the sugar nucleotide donor GDP-fucose was only recently detected in T. brucei, and the importance of fucose metabolism in this organism is not known. In this paper, we identified the genes encoding functional GDP-fucose biosynthesis enzymes in T. brucei and created conditional null mutants of TbGMD, the gene encoding the first enzyme in the pathway from GDP-mannose to GDP-fucose, in both bloodstream form and procyclic form parasites. Under nonpermissive conditions, both life cycle forms of the parasite became depleted in GDP-fucose and suffered growth arrest, demonstrating that fucose metabolism is essential to both life cycle stages. In procyclic form parasites, flagellar detachment from the cell body was also observed under nonpermissive conditions, suggesting that fucose plays a significant role in flagellar adhesion. Fluorescence microscopy of epitope-tagged TbGMD revealed that this enzyme is localized in glycosomes, despite the absence of PTS-1 or PTS-2 target sequences.


Comparative Evaluation of the LAMP Assay and PCR-Based Assays for the Rapid Detection of Alternaria solani.
Early blight (EB), caused by the pathogen Alternaria solani, is a major threat to global potato and tomato production. Early and accurate diagnosis of this disease is therefore important. In this study, we conducted a loop-mediated isothermal amplification (LAMP) assay, as well as conventional polymerase chain reaction (PCR), nested PCR, and quantitative real-time PCR (RT-qPCR) assays to determine which of these techniques was less time consuming, more sensitive, and more accurate. We based our assays on sequence-characterized amplified regions of the histidine kinase gene with an accession number (FJ424058). The LAMP assay provided more rapid and accurate results, amplifying the target pathogen in less than 60 min at 63°C, with 10-fold greater sensitivity than conventional PCR. Nested PCR was 100-fold more sensitive than the LAMP assay and 1000-fold more sensitive than conventional PCR. qPCR was the most sensitive among the assays evaluated, being 10-fold more sensitive than nested PCR for the least detectable genomic DNA concentration (100 fg). The LAMP assay was more sensitive than conventional PCR, but less sensitive than nested PCR and qPCR; however, it was simpler and faster than the other assays evaluated. Despite of the sensitivity, LAMP assay provided higher specificity than qPCR. The LAMP assay amplified A. solani artificially, allowing us to detect naturally infect young potato leaves, which produced early symptoms of EB. The LAMP assay also achieved positive amplification using diluted pure A. solani culture instead of genomic DNA. Hence, this technique has greater potential for developing quick and sensitive visual detection methods than do other conventional PCR strategies for detecting A. solani in infected plants and culture, permitting early prediction of disease and reducing the risk of epidemics.


OSlihc: An Online Prognostic Biomarker Analysis Tool for Hepatocellular Carcinoma.
Liver hepatocellular carcinoma (LIHC) is one of the most common malignant tumors in the world with an increasing number of fatalities. Identification of novel prognosis biomarker for LIHC may improve treatment and therefore patient outcomes. The availability of public gene expression profiling data offers the opportunity to discover prognosis biomarkers for LIHC. We developed an online consensus survival analysis tool named OSlihc using gene expression profiling and long-term follow-up data to identify new prognosis biomarkers. OSlihc consists of 637 cases from four independent cohorts. As a risk assessment tool, OSlihc generates the Kaplan-Meier survival plot with hazard ratio (HR) and p value to evaluate the prognostic value of a gene of interest. To test the reliability of OSlihc, we analyzed 65 previous reported prognostic biomarkers in OSlihc and showed that all of which have significant prognostic values. Furthermore, we identified four novel potential prognostic biomarkers (ATG9A, WIPI1, CXCL1, and CSNK2A2) for LIHC, the elevated expression of which predict the unfavorable survival outcomes. These genes (ATG9A, WIPI1, CXCL1, and CSNK2A2) may be potentially new biomarkers to identify at-risk LIHC patients when further validated. By OSlihc, users can evaluate the prognostic abilities of genes of their interest, which provides a platform for researchers to identify prognostic biomarkers to further develop targeted therapy strategies for LIHC patients. OSlihc is public and free to the users at http://bioinfo.henu.edu.cn/LIHC/LIHCList.jsp.


[Pre-operative care for liver disease patients].
Patients with impaired hepatic functional reserve when submitted to surgeries may have high rates of morbidity and mortality. Pre-existing liver disease should be detected without need for invasive methods. Clinical history and physical examination provide important clues. Laboratory liver function is not necessary unless there are changes in history or physical examination. Liver disease has many effects on surgery and anesthesia. A decrease in oxygenation and increased risk of liver dysfunction can be caused by anesthesia, hemorrhage, hypoxemia, hypotension, vasoactive drugs or the patient's position on the operating table during and after surgery. Emergency surgery is a major predictor of poor prognosis as well as sepsis and reoperations. The nature of liver disease, severity and type of surgery to be performed should take into account for a correct preoperative preparation. Some actions must be taken at preoperative to decrease chances of complications in patients with liver disease undergoing surgical procedures. Very close attention should be given to coagulopathy, encephalopathy, ascites, renal and pulmonary dysfunction, spontaneous bacterial peritonitis and esophageal varices. Patients with Child-Pugh score C and MELD>15 should not undergo elective surgery. Patients with Child-Pugh score B and MELD 10 to 15 may undergo minor surgical procedures with care in cases of extreme necessity. Patients with Child-Pugh score A and MELD<10 may be submitted to elective surgery.


[Pharmacological bases of coffee nutrients for diabetes prevention].
With an increasing number of studies describing the negative correlation of coffee consumption and the risk for type 2 diabetes mellitus, we were compelled to elucidate the nutrients which bring pharmacological effects on risk reduction for diabetes. In this review, the author's interest is focused on chlorogenic and caffeic acids derived from lightly roasted coffee beans, as well as nicotinic acid, volatile Maillard reaction products (vMRPs), and another structurally unknown compound contained in heavily roasted beans. Caffeine is a common compound in both lightly and heavily roasted beans and its anti-inflammatory effects on degenerative diseases such as diabetes mellitus has been reevaluated recently. The prophylactic effects of coffee on diabetes involve pleiotropy of plural components in accordance to the degree of the roasting. A new concept of nutritional blended coffee may be important to optimize the prophylactic effects of coffee on lowering the risk factors of diabetes and delaying the progress of diabetes complications as well.


Evaluation of risk and protective factors for work-related bite injuries to veterinary technicians certified in Minnesota.
To identify risk and protective factors for work-related bite injuries among veterinary technicians certified in Minnesota. Nested case-control study. 868 certified veterinary technicians (CVTs). A questionnaire was mailed to CVTs who previously participated in a survey regarding work-related injuries and did (cases; 301 surveys sent) or did not (controls; 567) report qualifying work-related animal bite injuries in the preceding 12 months. Descriptive statistics were summarized. Demographic and work-related variables for the month preceding the bite injury (for cases) or a randomly selected month (controls) were assessed with univariate analysis (489 CVTs) and multivariate analysis of a subset of 337 CVTs who worked in small or mixed mostly small animal facilities. Responses were received from 176 case and 313 control CVTs. For the subset of 337 CVTs, risk of bite injury was higher for those < 25 years of age (OR, 3.82; 95% confidence interval [CI], 1.84 to 7.94) than for those ≥ 35 years of age, for those who had worked < 5 years (OR, 3.24; 95% CI, 1.63 to 6.45) versus ≥ 10 years in any veterinary facility, and for those who handled ≥ 5 species/d (OR, 1.99; 95% CI, 1.06 to 3.74) versus < 3 species/d. Risk was lower for CVTs who handled < 10 versus ≥ 20 animals/d (OR, 0.23; 95% CI, 0.08 to 0.71). Several work-related factors were associated with the risk of work-related bite injury to CVTs. These findings may serve as a basis for development of intervention efforts and future research regarding work-related injuries among veterinary staff.


Autologous circulating angiogenic cells treated with osteopontin and delivered via a collagen scaffold enhance wound healing in the alloxan-induced diabetic rabbit ear ulcer model.
Diabetic foot ulceration is the leading cause of amputation in people with diabetes mellitus. Peripheral vascular disease is present in the majority of patients with diabetic foot ulcers. Despite standard treatments there exists a high amputation rate. Circulating angiogenic cells previously known as early endothelial progenitor cells are derived from peripheral blood and support angiogenesis and vasculogenesis, providing a potential topical treatment for non-healing diabetic foot ulcers. A scaffold fabricated from Type 1 collagen facilitates topical cell delivery to a diabetic wound. Osteopontin is a matricellular protein involved in wound healing and increases the angiogenic potential of circulating angiogenic cells. A collagen scaffold seeded with circulating angiogenic cells was developed. Subsequently the effect of autologous circulating angiogenic cells that were seeded in a collagen scaffold and topically delivered to a hyperglycemic cutaneous wound was assessed. The alloxan-induced diabetic rabbit ear ulcer model was used to determine healing in response to the following treatments: collagen seeded with autologous circulating angiogenic cells exposed to osteopontin, collagen seeded with autologous circulating angiogenic cells, collagen alone and untreated wound. Stereology was used to assess angiogenesis in wounds. The cells exposed to osteopontin and seeded on collagen increased percentage wound closure as compared to other groups. Increased angiogenesis was observed with the treatment of collagen and collagen seeded with circulating angiogenic cells. These results demonstrate that topical treatment of full thickness cutaneous ulcers with autologous circulating angiogenic cells increases wound healing. Cells exposed to the matricellular protein osteopontin result in superior wound healing. The wound healing benefit is associated with a more efficient vascular network. This topical therapy provides a potential novel therapy for the treatment of non-healing diabetic foot ulcers in humans.


The role of radionuclide therapy in medullary thyroid cancer.
In medullary thyroid carcinoma (MTC) the detection of occult metastases is difficult and the prognosis of widespread disease is poor. In recent years several radiopharmaceuticals have become available for the diagnosis of this tumor. None of these tracers, however, has satisfactory diagnostic sensitivity and specificity. Furthermore, only few radiopharmaceutical compounds proved to have clinical value in therapeutic applications. Radionuclide therapy utilizes unsealed radioactive sources in order to deliver selective irradiation to the target organs or cancer lesions. This approach is only clinically indicated when there is a scintigraphic evidence of sufficient tumor uptake and a favorable biodistribution. When these conditions are present, radionuclide therapy can be adopted in MTC patients. Due to the low incidence of this tumor, the poor sensitivity of the available radiopharmaceuticals and their limited indications, the clinical experience in radionuclide therapy of MTC is still limited and there is general agreement among experts that it has only a palliative role. Here we briefly report the main experiences in radionuclide therapy in the past and in recent years. In addition, we summarize the results obtained with 131I-MIBG therapy at the Istituto Nazionale Tumori of Milan, as well as the most important ongoing preclinical and phase I/II trials.


A prospective study of individual-level social capital and major depression in the United States.
To investigate prospectively the associations between depression and cognitive social capital (social trust, sense of belonging, mutual aid) and structural social capital (volunteer work and community participation). This was a prospective study that was carried out in the USA. The participants were a nationally representative sample of 724 English-speaking non-institutionalised adults (25-74 years old) who participated in the National Survey of Midlife Development in the United States (MIDUS) in 1995-6 and the MIDUS Psychological Experience Follow-Up study in 1998. In multivariable adjusted logistic regression analyses, those who trusted their neighbours were less likely to develop major depression (MD) during follow-up than those who reported low levels of social capital on these dimensions (adjusted OR of MD for high vs low trust = 0.43; 95% CI 0.20 to 0.93, adjusted for MD at baseline, age, gender, race, education, working status, marital status, physical health and extroversion traits). Structural dimensions of social capital were not associated with MD in adjusted models. Perceptions of higher levels of cognitive social capital (trust of neighbours) are associated with lower risks of developing MD during 2-3 year follow-up. However, after excluding participants with MD at the baseline, the association between trust and MD became non-significant. Structural dimensions were not associated with MD.


Antibodies to DNA repair proteins in headache with neurological deficits and cerebrospinal fluid lymphocytosis (HaNDL) patients.
Autoimmune mechanisms have been implicated in the pathogenesis of headache and neurological deficits with cerebrospinal fluid lymphocytosis (HaNDL). Pooled sera of five HaNDL patients and 30 controls (10 multiple sclerosis patients, 10 migraine patients, 10 healthy controls) were screened by protein macroarray. All sera were also individually subjected to immunoprecipitation with neuroblastoma cells and the bound antigens were identified by mass spectrometry. Antibodies to three DNA repair proteins (mitogen-activated protein kinase-4, DNA-dependent protein kinase catalytic subunit, DNA excision repair protein ERCC-6) were identified by both macroarray and immunoprecipitation methods in 3/5 HaNDL sera, but in none of the controls. The presence of DNA repair protein antibodies indicates DNA damage and provides further support for the inflammatory etiology of HaNDL.


Identification of a safe and adequate division point of the left-sided bile duct with magnetic resonance cholangiography during donor left lateral sectionectomy.
Multiple bile duct (BD) openings on the graft can cause postoperative BD-related complications as the result of their small orifices and multiple anastomoses. This study aimed to determine a safe and adequate BD division point during donor left lateral sectionectomy. Left BD was classified into type I: B4 enters the common trunks of B2 and B3; type II: B2 joins the common channels of B3 and B4; or type III: B2, B3, and B4 join. We assessed the chance of multiple openings with 3 BD types in 43 left lateral living donor grafts from January 2004 to January 2011. We also analyzed the potential for multiple openings and right-sided BD injury, based on BD types, during left lateral sectionectomy according to the distance from the crossing point of the right lateral border of the umbilical portion of the left portal vein at the left BD (point U) via preoperative magnetic resonance imaging from another 90 living donors who underwent several types of hepatectomy from January 2012 to December 2012. There was a statistically marginal relevance for multiple openings between BD type I and type II (33.3% in type II vs 7.4% in type I, P = .078), and the optimal BD division points were different by BD types (right-side 5 mm from point U in type I; 10 mm in type II; and 12 mm in type III). The safe and adequate BD division point should be determined according to the left BD types during living donor left lateral sectionectomy.


Left atrial infarction: a case report and review of the literature.
The majority of cardiac related deaths are due to ischemic heart disease, with the most common clinical scenario being severe coronary artery atherosclerosis resulting in left ventricular myocardial infarction. However, infarction of other cardiac chambers does occur, and often has specific clinical associations. We report a case of a 70-year-old man who suffered from left atrial infarction that resulted in a transmural rupture of his left atrium. The patient had a history of rheumatic heart disease, mitral valve stenosis, and severe atherosclerotic coronary artery disease. Four days before death, he underwent mitral valve replacement and left circumflex coronary artery bypass. Two days later, he developed atrial fibrillation. On the day of death, he had decreased mental status, questionable seizure activity, hematemesis, ventricular tachycardia, and eventually asystole. At autopsy, he had significant hemopericardium with a fibrinous pericarditis and bilateral hemothoraces (total blood volume: 1250 mL). A 0.1 to 0.2 cm left atrial transmural defect was identified. The prosthetic mitral valve was free of vegetations, and completely intact. Similarly, the left circumflex artery bypass graft was completely patent and unremarkable. Severe calcific atherosclerosis was of his native left circumflex and left main coronary arteries. Microscopic examination revealed acute myocardial infarction of the left atrium at the rupture site. The anatomy of atrial circulation as well as the pathology and consequences of atrial infarction are discussed.


Omega-6 fatty acids and inflammation.
Inflammation is a normal process that is part of host defence and tissue healing. However, excessive or unresolved inflammation can lead to uncontrolled tissue damage, pathology and disease. In humans on a Western diet, the omega-6 polyunsaturated fatty acid arachidonic acid (ARA) makes a significant contribution to the fatty acids present in the membrane phospholipids of cells involved in inflammation. ARA is a precursor to a number of potent pro-inflammatory mediators including well described prostaglandins and leukotrienes, which has led to the development of anti-inflammatory pharmaceuticals that target the ARA pathway to successfully control inflammation. Hence, it is commonly believed that increasing dietary intake of the omega-6 fatty acids ARA or its precursor linoleic acid (LA) will increase inflammation. However, studies in healthy human adults have found that increased intake of ARA or LA does not increase the concentrations of many inflammatory markers. Epidemiological studies have even suggested that ARA and LA may be linked to reduced inflammation. Contrastingly, there is also evidence that a high omega-6 fatty acid diet inhibits the anti-inflammatory and inflammation-resolving effect of the omega-3 fatty acids. Thus, the interaction of omega-3 and omega-6 fatty acids and their lipid mediators in the context of inflammation is complex and still not properly understood.


A randomised non-comparative phase II trial of cixutumumab (IMC-A12) or ramucirumab (IMC-1121B) plus mitoxantrone and prednisone in men with metastatic docetaxel-pretreated castration-resistant prostate cancer.
Cixutumumab, a human monoclonal antibody (HuMAb), targets the insulin-like growth factor receptor. Ramucirumab is a recombinant HuMAb that binds to vascular endothelial growth factor receptor-2. A non-comparative randomised phase II study evaluated cixutumumab or ramucirumab plus mitoxantrone and prednisone (MP) in metastatic castration-resistant prostate cancer (mCRPC). Men with progressive mCRPC during or after docetaxel therapy received mitoxantrone 12 mg/m(2) on day 1 and prednisone 5mg twice daily and were randomised 1:1 to receive either cixutumumab or ramucirumab 6 mg/kg intravenously weekly in a 21-day cycle. Primary end-point was composite progression-free survival (cPFS). Secondary end-points included safety, response, radiographic progression-free survival (PFS) and overall survival (OS). Sample size was based on a 50% increase in median cPFS from 2.6 (MP) to 3.9 months (either combination). 132 men were treated (66 per arm). Median cPFS was 4.1 months (95% confidence interval (CI), 2.2-5.6) for cixutumumab and 6.7 months (95% CI, 4.5-8.3) for ramucirumab. Median time to radiographic progression was 7.5 months for cixutumumab and 10.2 months for ramucirumab, with a median OS of 10.8 and 13.0 months, respectively. Fatigue was the most frequent adverse event (AE). Incidence of most non-haematologic grade 3-4 AEs was <10% on both arms. Grade 3 cardiac dysfunction occurred in 7.6% of patients on ramucirumab. Combinations of cixutumumab or ramucirumab plus MP were feasible and associated with moderate toxicities in docetaxel-pretreated men with mCRPC. Of the two regimens, the ramucirumab regimen is worthy of further testing based on the observed cPFS relative to the historical control.


A finite element model for simulating acoustic streaming in cystic breast lesions with experimental validation.
Streaming detection is an ultrasonic technique that can be used to distinguish fluid-filled lesions, or cysts, from solid lesions. With this technique, high intensity ultrasound pulses are used to induce acoustic streaming in cyst fluid, and this motion is detected using Doppler flow estimation methods. Results from a pilot clinical study were recently published in which acoustic streaming was successfully induced and detected in 14 of 15 simple breast cysts and four of 14 sonographically indeterminate breast lesions in vivo. In the study, the detected velocities were found to vary considerably among cysts and for different pulsing regimes. A finite element model of streaming detection is presented. This model is utilized to investigate methods of increasing induced acoustic streaming velocity while minimizing patient exposure to high intensity ultrasound during streaming detection. Parameters studied include intensity, frequency, acoustic beam shape, cyst-diameter, cyst fluid protein concentration, and cyst fluid viscosity. The model, which provides both transient and steady-state solutions, is shown to predict trends in streaming velocity accurately. Experimental results from studies investigating the potential for nonlinear streaming enhancement in cysts are also provided.


Microinjection of Cre recombinase protein into zygotes enables specific deletion of two eukaryotic selection cassettes and enhances the expression of a DsRed2 reporter gene in Ccr2/Ccr5 double-deficient mice.
The chemokine receptors CCR2 and CCR5 represent potential novel therapeutic targets to treat important inflammatory and infectious diseases, including atherosclerosis and HIV infection. To study the functions of both receptors in vivo, we aimed to generate Ccr2/Ccr5 double-deficient mice. As these genes are separated by <20 kb, they were inactivated consecutively by two rounds of gene targeting in embryonic stem (ES) cells. Thereby neomycin and hygromycin selection cassettes flanked by four identical loxP recognition sequences for Cre recombinase were integrated into the ES cell genome together with EGFP and DsRed2 reporter genes. Both selection cassettes could be deleted in vitro by transiently transfecting ES cells with Cre expression vectors. However, after blastocyst microinjection these cells yielded only weak chimeras, and germline transmission was not achieved. Therefore, Ccr2/Ccr5 double-deficient mice were generated from ES cells still carrying both selection cassettes. Microinjection of zygotes with a recombinant fusion protein consisting of maltose-binding protein and Cre (MBP-Cre) allowed the selective deletion of both cassettes. All sequences in between and both reporter genes were left intact. Deletion of both selection cassettes resulted in enhanced DsRed2 reporter gene expression. Cre protein microinjection of zygotes represents a novel approach to perform complex recombination tasks.


[Utilization of inorganic sulfur sources by Staphylococcus aureus strains].
Staphylococcus aureus strains of different host-adapted variants (Meyer 1966) have been tested for their ability to use inorganic sulfur sources. All the 25 strains tested were able to utilize sodium sulfide as sulfur source in a medium similar to that described by Kloos and Pattee (1965). Using S. aureus strain 116/74 grown in a medium containing Na2-35S as the only sulfur source we studied incorporation and insertion of inorganic sulfide into sulfur containing amino acids. In disintegrated and fractionated cellular material we could find 35S labelled homocystine and methionine as major compounds, and cystine, cysteic acid, homocysteic acid, and beta-sulphopyruvate as minor compounds. The occurrence of homocystine and the sulfonic acids in bacterial proteins is rather uncommon.


Potential of Aqueous Humor as a Surrogate Tumor Biopsy for Retinoblastoma.
Retinoblastoma (Rb) is one of the first tumors to have a known genetic etiology. However, because biopsy of this tumor is contraindicated, it has not been possible to define the effects of secondary genetic changes on the disease course. To investigate whether the aqueous humor (AH) of Rb eyes has sufficient tumor-derived DNA to perform genetic analysis of the tumor, including DNA copy number alterations. This investigation was a case series study at a tertiary care hospital (Children's Hospital Los Angeles) with a large Rb treatment center. Cell-free DNA (cfDNA) was isolated from 6 AH samples from 3 children with Rb, including 2 after primary enucleation and 1 undergoing multiple intravitreous injections of melphalan for vitreous seeding. Samples were taken between December 2014 and September 2015. Measurable levels of nucleic acids in the AH and identification of tumor-derived DNA copy number variation in the AH. The AH was analyzed for DNA, RNA, and micro-RNA using Qubit high-sensitivity kits. Cell-free DNA was isolated from the AH, and sequencing library protocols were optimized. Shallow whole-genome sequencing was performed on an Illumina platform, followed by genome-wide chromosomal copy number variation profiling to assess the presence of tumor DNA fractions in the AH cfDNA of the 3 patients. One child's cfDNA from the AH and tumor DNA were subjected to Sanger sequencing to isolate the RB1 mutation. Six AH samples were obtained from 3 Rb eyes in 3 children (2 male and 1 female; diagnosed at ages 7, 20, and 28 months). A corroborative pattern between the chromosomal copy number variation profiles of the AH cfDNA and tumor-derived DNA from the enucleated samples was identified. In addition, a nonsense RB1 mutation (Lys→STOP) from 1 child was also identified from the AH samples obtained during intravitreous injection of melphalan, which matched the tumor sample postsecondary enucleation. Sanger sequencing of the AH cfDNA and tumor DNA with polymerase chain reaction primers targeting RB1 gene c.1075A demonstrated this same RB1 mutation. In this study evaluating nucleic acids in the AH from Rb eyes undergoing salvage therapy with intravitreous injection of melphalan, the results suggest that the AH can serve as a surrogate tumor biopsy when Rb tumor tissue is not available. This novel method will allow for analyses of tumor-derived DNA in Rb eyes undergoing salvage therapy that have not been enucleated.


Dominant effect of protein charge rather than protein shape in apparent DNA bending by engineered bZIP domains.
We are interested in how asymmetric charge neutralization of DNA by proteins results in DNA bending. We have previously reported electrophoretic phasing experiments utilizing homodimer peptides derived from GCN4, a yeast basic zipper (bZIP) transcription factor. Here we report the results of experiments that examine the importance of peptide sequence context in DNA bending and test the hypothesis that peptide structural asymmetry causes electrophoretic anomalies in the absence of DNA bending. We prepared two new series of bZIP peptides that differed dramatically in overall size, structure, and peptide sequence near the DNA. The magnitude of apparent DNA bending is independent of the structure of the protein. This result reduces the concern that bZIP protein structure causes electrophoretic anomalies in the absence of DNA bending. In all cases, both the magnitude and direction of the apparent DNA bend angle are strongly dependent on the local peptide charge. We attempted to validate independently our results with a minicircle competition binding assay. Binding preferences of severalfold for properly phased circular versus linear DNA templates were predicted. However, no binding preferences were observed. We propose that the minicircle binding assay may be intrinsically insensitive to DNA bending or flexibility induced by the bZIP peptides studied, and we provide a unifying explanation for the discrepancies between the cyclization and electrophoretic experiments.


Symmetry in computed tomography of the brain: the pitfalls.
Computed tomography (CT) studies of the brain are one of the most frequent examinations interpreted by radiologists out of hours. Apparently normal appearances in patients with significant neurological morbidity can be perplexing. As the contents of the cranium are normally remarkably symmetrical on axial CT, disease entities that result in symmetrical appearances are the most difficult to detect. In this review we highlight a spectrum of important acute neurological conditions that result in abnormal but symmetrical appearances on CT.


Men in nursing: making the right choice.
Why do men choose nursing as an occupation? What are the perceived barriers for men working in a predominately female-oriented profession? To answer these questions, the authors conducted a study in 2005, using a self-report survey as the data collecting tool. The 250 male registered nurses in Newfoundland and Labrador were the target population. The most common reasons for entering nursing cited by the respondents (N = 62) were career opportunities, job security and salary. The most commonly perceived barriers were sexual stereotypes, lack of recruitment strategies, female-oriented profession and lack of exposure to male role models in the media. The respondents stressed that recruitment strategies should focus on the factors that are important to them: job security, career opportunities and salary. The findings from this research may help with future recruitment strategies designed to attract more men into the nursing profession.


Differential species sensitivity to the inhibitory effect of cardiac glycosides on 3H-1-noradrenaline accumulation by tissue slices.
Effects of three cardiac glycosides on the accumulation of 3H-1-noradrenaline (3H-1-NA) by slices of heart and spleen were studied. Ouabain, digitoxin and digoxin, all produced a concentration dependent inhibition of 3H-1-NA uptake in both types of tissue slices. The maximum amount of 3H-1-NA accumulated as well as the rate of uptake was decreased. Digitoxin and ouabain were equipotent; however, digoxin was significantly less potent. Tissues from different mamalian specis did not exhibit the same degree of sensitivity to the inhibitory effect of cardiac glycosides on 3H-1-NA accumulation. Dogs were most sensitive and guinea-pigs an order of magnitude less sensitive. Rats were least sensitive by roughly two orders of magnitude when compared with guinea-pigs. The relationship of the effect of digitalis on 3H-1-NA accumulation to digitalis-induced cardiac arrhythmias is discussed. Finally, the pattern of species sensitivity found here is compared with that observed in relation to inhibition of Na+-K+-ATPase by cardiac glycosides.


Bovine milk fat globule membrane proteome.
Milk fat globule membranes (MFGM) were isolated from the milk of mid-lactation Holstein cows. The purified MFGM were fractionated using 1-dimensional SDS gels. Tryptic peptides from gel slices were further fractionated on a micro-capillary high performance liquid chromatograph connected to a nanospray-tandem mass spectrometer. Analysis of the data resulted in 120 proteins being identified by two or more unique peptide sequences. Of these 120 proteins, 71% are membrane associated proteins with the remainder being cytoplasmic or secreted proteins. Only 15 of the proteins identified in the cow MFGM were the same as proteins identified in previous mouse or human MFGM proteomic studies. Thus, the bulk of the proteins identified are new for bovine MFGM proteomics. The proteins identified were associated with membrane/protein trafficking (23%), cell signalling (23%), unknown functions (21%), fat transport/metabolism (11%), transport (9%), protein synthesis/folding (7%), immune proteins (4%) and milk proteins (2%). The proteins associated with cell signalling or membrane/protein trafficking may provide insights into MFGM secretion mechanisms. The finding of CD14, toll like receptor (TLR2), and TLR4 on MFGM suggests a direct role for the mammary gland in detecting an infection.


Production of the long pentraxin PTX3 in advanced atherosclerotic plaques.
Elevated plasma levels of the pentraxin protein family member C-reactive protein (CRP) are associated with increased risk of cardiovascular disease in both healthy and high-risk subjects. The long pentraxin family member, pentraxin 3 (PTX3), was recently described. Like CRP, PTX3 is induced by acute inflammatory stimuli and is increased in the blood of patients with acute myocardial infarction. Unlike CRP, it is expressed in a wide range of cell types, but not in hepatocytes. In this study, we have investigated the expression of PTX3 in atherosclerosis. Immunohistochemical staining of advanced atherosclerotic lesions revealed strong expression of PTX3. In contrast, no PTX3 expression was observed in nonatherosclerotic internal mammary arteries. By staining serial sections with cell type- and PTX3-specific antibodies, we observed that PTX3 was produced principally by macrophages and endothelial cells. Infrequent expression by smooth muscle cells was also observed. Our results suggest that PTX3 may contribute to the pathogenesis of atherosclerosis.


Non-developmental dimensions of adult regeneration in Hydra.
An essential dimension of 3D regeneration in adult animals is developmental, with the formation of organizers from somatic tissues. These organizers produce signals that recruit surrounding cells and drive the restoration of the missing structures (organs, appendages, body parts). However, even in animals with a high regenerative potential, this developmental potential is not sufficient to achieve regeneration as homeostatic conditions at the time of injury need to be "pro-regenerative". In Hydra, we identified four distinct homeostatic properties that provide a pro-regenerative framework and we discuss here how these non-developmental properties impact regeneration. First, both the epithelial and the interstitial-derived cells are highly plastic along the animal body, a plasticity that offers several routes to achieve regeneration. Second, the abundant stocks of continuously self-renewing adult stem cells form a constitutive pro-blastema in the central body column, readily activated upon bisection. Third, the autophagy machinery in epithelial cells guarantees a high level of fitness and adaptation to detrimental environmental conditions, as evidenced by the loss of regeneration in animals where autophagy is dysfunctional. Fourth, the extracellular matrix, named mesoglea in Hydra, provides a dynamically-patterned environment where the molecular and mechanical signals induced by injury get translated into a regenerative process. We claim that these homeostatic pro-regenerative features contribute to define the high regenerative potential of adult Hydra.


[Effects of cisapride on the motility of the digestive tract in dogs and guinea pigs].
Effects of cisapride on the motility of the digestive tract in vivo in dogs and the guinea-pig intestine in vitro were studied. Cisapride (0.05-2.0 mg/kg, i.v.) produced an increase in amplitude of spontaneous contractions and basal tone in the stomach, duodenum, jejunum and proximal and distal colon in dogs. In some animals, however, it induced an inhibition with decrease in amplitude and tone. It also induced an increase in amplitude of contractions in the gallbladder and the sphincter of Oddi in dogs. The tone of the gallbladder was elevated by the same dose of cisapride, but the tone of the sphincter of Oddi was decreased. The drug produced a reverse response in some animals. These excitatory responses to cisapride were abolished by atropine (0.2 mg/kg, i.v.). Motility of the guinea-pig isolated ileum and colon was enhanced with an increase in their amplitude of contractions and basal tone at low concentrations of cisapride (10(-9)-10(-6)M) but it was inhibited at higher concentrations (10(-5)-10(-4)M). Atropine abolished the excitatory response of the ileum to cisapride in all cases. It abolished the excitation of the colon in some preparations but reduced only in some degree in the other. The inhibitory effect of cisapride on isolated preparations was unaffected by tetrodotoxin. From these results, it is concluded that cisapride enhances motility of the gastrointestinal tract and biliary tract by acting on myenteric cholinergic neurons and inhibits it by acting on the smooth muscle itself.


Long non-coding RNA TCF7 predicts the progression and facilitates the growth and metastasis of colorectal cancer.
Long non-coding RNA (lnc)TCF7 has been reported to promote the self‑renewal of human cancer stem cells, and enhance the aggressiveness of human non‑small cell lung cancer and hepatocellular carcinoma cells. However, the effect of lncTCF7 on colorectal cancer (CRC) tumorigenesis and progression is currently unclear. In the present study, reverse transcription‑quantitative polymerase chain reaction results demonstrated that lncTCF7 expression was higher in CRC tissues compared with adjacent normal tissues and was significantly associated with tumor size, differentiation degree, tumor‑node‑metastasis grade, lymph node metastasis and invasion depth. In addition, lncTCF7 demonstrated a high sensitivity and specificity for diagnosing CRC, as determined by receiver operating characteristic curve analysis. Furthermore, lncTCF7 silencing in SW‑620 and HT29 CRC cell lines inhibited the proliferation, cell cycle, migration and invasion of cells, as determined by Cell Counting Kit‑8 assays, propidium iodide (PI) staining and flow cytometry, wound healing assays and Transwell invasion assays, respectively; however, Annexin V/PI double staining and flow cytometry indicated that lncTCF7 silencing did not significantly affect the apoptosis of CRC cells. These results indicate that lncTCF7 may predict the progression, and promote the growth and metastasis, of CRC, and may therefore be a novel diagnostic marker and therapeutic target for CRC treatment.