Alpha-synuclein in Lewy body disease and Alzheimer's disease.
Alzheimer's disease (AD) and Lewy body disease (LBD) are the most common causes of dementia in the elderly population. Previous studies have shown that cognitive alterations in these disorders are associated with synaptic loss. Injury and loss of synapses might be associated with altered function of synaptic proteins. Among them, recent studies have shown that abnormal aggregation and accumulation of synaptic proteins, such as alpha-synuclein, might be associated with plaque formation in AD and Lewy body formation in LBD. Further reinforcing the hypothesis that alpha-synuclein plays a major role in the pathogenesis of these disorders, recent work has shown that mutations that alter the conformation of this molecule are associated with familial forms of Parkinson's disease. The mechanisms by which altered function or aggregation of alpha-synuclein might lead to neurodegeneration are not completely clear; however, new evidence points to a potential role for this molecule in synaptic damage and neurotoxicity via amyloid-like fibril formation and mitochondrial dysfunction. In this manuscript we review the data linking alpha-synuclein to the pathogenesis of AD and LBD.


Enhanced cationic liposome-mediated transfection using the DNA-binding peptide mu (mu) from the adenovirus core.
Promising advances in nonviral gene transfer have been made as a result of the production of cationic liposomes formulated with synthetic cationic lipids (cytofectins) that are able to transfect cells. However few cationic liposome systems have been examined for their ability to transfect CNS cells. Building upon our earlier use of cationic liposomes formulated from 3beta-[N-(N',N'-dimethylaminoethane)carbamoyl] cholesterol (DC-Chol) and dioleoyl-L-alpha-phosphatidyl-ethanolamine (DOPE), we describe studies using two cationic viral peptides, mu (mu) and Vp1, as potential enhancers for cationic liposome-mediated transfection. Mu is derived from the condensed core of the adenovirus and was selected to be a powerful nucleic acid charge neutralising and condensing agent. Vp1 derives from the polyomavirus and harbours a classical nuclear localisation signal (NLS). Vp1 proved disappointing but lipopolyplex mixtures formulated from pCMVbeta plasmid, mu peptide and DC-Chol/DOPE cationic liposomes were able to transfect an undifferentiated neuronal ND7 cell line with beta-galactosidase reporter gene five-fold more effectively than lipoplex mixtures prepared from pCMVbeta plasmid and DC-Chol/DOPE cationic liposomes. Mu was found to give an identical enhancement to cationic liposome-mediated transfection of ND7 cells as poly-L-lysine (pLL) or protamine sulfate (PA). The enhancing effects of mu were found to be even greater (six- to 10-fold) when differentiated ND7 cells were transfected with mu-containing lipopolyplex mixtures. Differentiated ND7 cells represent a simple ex vivo-like post-mitotic CNS cell system. Successful transfection of these cells bodes well for transfection of primary neurons and CNS cells in vivo. These findings have implications for experimental and therapeutic uses of cationic liposome-mediated delivery of nucleic acids to CNS cells.


Factors associated with Postpartum Depression and Its Influence on Maternal Identity.
This study aimed to examine the factors associated with postpartum depression and its influence on maternal identity of postpartum women. Research design was a cross sectional descriptive study with a total of 89 women within the six month postpartum period. Associations of eating habits, overall sleep quality and other factors with postpartum depression utilizing the Korean Beck Depression Inventory (K-BDI) were done. The influence of postpartum depression on maternal identity was analyzed. Variables yielding significant associations (p<.05) were included in an adjusted logistic regression and a stepwise multiple regression. Mean scores of postpartum depression was 9.42+/-6.08 and 31.5% (n=28) for mild depression, 11.2% (n=10) was moderate and 4.5% (n=4) was severe depression on the K-BDI scale. Perceived health status and overall sleep quality were predictors of postpartum depression. Postpartum depression and the husband's love were predictors of maternal identity. Awareness of poor health perception and sleep quality will be helpful to detect for postpartum depression. Strategies to increase maternal identity during the postpartum period would be tailored by level of depression.


Therapeutic applications of sustainable new chitosan derivatives and its nanocomposites: Fabrication and characterization.
Chitosan (CS) is a biologically active biopolymer used in different medical applications due to its biodegradability, biocompatibility, and nontoxicity. Nanotechnology is an exciting and quick developing field in medical applications. Nanoparticles have shown great potential in the treatment of cancer and inflammation. In the present work modification of chitosan and its (Ag, Au, or ZnO) nanocomposites by N-aminophthalimide (NAP) occurred through the reaction with epichlorohydrin (ECH) as a crosslinker in the presence or absence of glutaraldehyde (GA) under different reaction conditions using microwave irradiation to give modified chitosan derivatives CS-2, CS-6, and their nanocomposites. Modified chitosan derivatives were characterized using different tools. CS-2 and CS-6 derivatives displayed enhancement of thermal stability and crystallinity compared to chitosan. Additionally, CS-2, CS-6, and their nanocomposites exhibited improvements in antitumor activity against HeLa cancer cells and enzymatic inhibitory against trypsin and α-chymotrypsin enzymes compared to chitosan. However, CS-2 revealed the highest cell growth inhibition% toward HeLa cells (89.02 ± 1.46 %) and the enzymatic inhibitory toward α-chymotrypsin enzyme (17.13 ± 1.59 %). Furthermore, CS-Au-2 showed the highest enzymatic inhibitory against trypsin enzyme (28.14 ± 1.76 %). These results suggested that the new chitosan derivatives CS-2, CS-6, and their nanocomposites could be a platform for medical applications against HeLa cells, trypsin, and α-chymotrypsin enzymes.


A Physiological-Based Pharmacokinetic Model Embedded with a Target-Mediated Drug Disposition Mechanism Can Characterize Single-Dose Warfarin Pharmacokinetic Profiles in Subjects with Various CYP2C9 Genotypes under Different Cotreatments.
Warfarin, a commonly prescribed oral anticoagulant medication, is highly effective in treating deep vein thrombosis and pulmonary embolism. However, the clinical dosing of warfarin is complicated by high interindividual variability in drug exposure and response and its narrow therapeutic index. CYP2C9 genetic polymorphism and drug-drug interactions (DDIs) are substantial contributors to this high variability of warfarin pharmacokinetics (PK), among numerous factors. Building a physiology-based pharmacokinetic (PBPK) model for warfarin is not only critical for a mechanistic characterization of warfarin PK but also useful for investigating the complicated dose-exposure relationship of warfarin. Thus, the objective of this study was to develop a PBPK model for warfarin that integrates information regarding CYP2C9 genetic polymorphisms and their impact on DDIs. Generic PBPK models for both S- and R-warfarin, the two enantiomers of warfarin, were constructed in R with the mrgsolve package. As expected, a generic PBPK model structure did not adequately characterize the warfarin PK profile collected up to 15 days following the administration of a single oral dose of warfarin, especially for S-warfarin. However, following the integration of an empirical target-mediated drug disposition (TMDD) component, the PBPK-TMDD model well characterized the PK profiles collected for both S- and R-warfarin in subjects with different CYP2C9 genotypes. Following the integration of enzyme inhibition and induction effects, the PBPK-TMDD model also characterized the PK profiles of both S- and R-warfarin in various DDI settings. The developed mathematic framework may be useful in building algorithms to better inform the clinical dosing of warfarin. SIGNIFICANCE STATEMENT: The present study found that a traditional physiology-based pharmacokinetic (PBPK) model cannot sufficiently characterize the pharmacokinetic profiles of warfarin enantiomers when warfarin is administered as a single dose, but a PBPK model with a target-mediated drug disposition mechanism can. After incorporating CYP2C9 genotypes and drug-drug interaction information, the developed model is anticipated to facilitate the understanding of warfarin disposition in subjects with different CYP2C9 genotypes in the absence and presence of both cytochrome P450 inhibitors and cytochrome P450 inducers.


Intranuclear localization of apoptosis-inducing factor (AIF) and large scale DNA fragmentation after traumatic brain injury in rats and in neuronal cultures exposed to peroxynitrite.
Programmed cell death occurs after ischemic, excitotoxic, and traumatic brain injury (TBI). Recently, a caspase-independent pathway involving intranuclear translocation of mitochondrial apoptosis-inducing factor (AIF) has been reported in vitro; but whether this occurs after acute brain injury was unknown. To address this question adult rats were sacrificed at various times after TBI. Western blot analysis on subcellular protein fractions demonstrated intranuclear localization of AIF in ipsilateral cortex and hippocampus at 2-72 h. Immunocytochemical analysis showed AIF labeling in neuronal nuclei with DNA fragmentation in the ipsilateral cortex and hippocampus. Immunoelectronmicroscopy verified intranuclear localization of AIF in hippocampal neurons after TBI, primarily in regions of euchromatin. Large-scale DNA fragmentation ( approximately 50 kbp), a signature event in AIF-mediated cell death, was detected in ipsilateral cortex and hippocampi by 6 h. Neuron-enriched cultures exposed to peroxynitrite also demonstrated intranuclear AIF and large-scale DNA fragmentation concurrent with impaired mitochondrial respiration and cell death, events that are inhibited by treatment with a peroxynitrite decomposition catalyst. Intranuclear localization of AIF and large-scale DNA fragmentation occurs after TBI and in neurons under conditions of oxidative/nitrosative stress, providing the first evidence of this alternative mechanism by which programmed cell death may proceed in neurons after brain injury.


Disparities among patients with respiratory failure.
Disparities are common within healthcare, and critical illness is no exception. This review summarizes recent literature on health disparities within respiratory failure, focusing on race, ethnicity, socioeconomic status, and sex. Current evidence indicates that Black patients have higher incidence of respiratory failure, while the relationships among race, ethnicity, and mortality remains unclear. There has been renewed interest in medical device bias, specifically pulse oximetry, for which data demonstrate patients with darker skin tones may be at risk for undetected hypoxemia and worse outcomes. Lower socioeconomic status is associated with higher mortality, and respiratory failure can potentiate socioeconomic inequities via illness-related financial toxicity. Literature on sex-based disparities is limited; however, evidence suggests males receive more invasive care, including mechanical ventilation. Most studies focused on disparities in incidence and mortality associated with respiratory failure, but few relied on granular clinical data of patients from diverse backgrounds. Future studies should evaluate processes of care for respiratory failure that may mechanistically contribute to disparities in order to develop interventions that improve outcomes.


Review of techniques for ultrasonic determination of kidney stone size.
Imaging is a vital cog in the wheel of diagnosis and management of patients suspected with renal and ureteric calculi, and it is imperative to choose the appropriate investigation that is accurate as well as safe for the patient. At present, computed tomography (CT) is the gold standard for a patient suspected to have stone disease. However, CT scan is associated with the hazards of radiation and high cost. Ultrasonography (US) is cheap and also devoid of any radiation hazard to the patient. But, at the same time, its usage is limited by decreased sensitivity and specificity, inaccuracy in measuring stone size, and observer dependency. In this article, we review the techniques to improve the accuracy of US in measuring stone size. According to a review, the sensitivity and specificity for renal calculi are 45% and 88%, respectively, and for ureteric calculi, they are 45% and 94%, respectively. The sensitivity of US decreased when the size of the stone is <3 mm and also in a nondilated system, and the sensitivity increased as the size of the stone increased. There are factors that can be adjusted to increase the accuracy of stone measurement. The main factors are changes in gain and depth and alternate modes such as flash angle imaging, harmonic mode, and S mode. Also measures such as use of shadow for size measurement can help in improving the accuracy of stone size measurement. A new automated computerized stone-sizing program improves the accuracy of stone size calculation and reduces user variability. US is an ideal first-line imaging modality for nephrolithiasis due to its advantages such as low cost, absence of radiation, and easy availability. However, the only limitation is its reduced sensitivity and specificity when compared with CT. The addition of newer modes can improve the accuracy in stone size measurement.


Mechanical evaluation of a carbonated apatite cement in the fixation of unstable intertrochanteric fractures.
We created three-part unstable intertrochanteric fractures in 6 pairs of aged, osteopenic, human, cadaveric femora. Fractures were reduced and fixed with a Dynamic Hip Screw (DHS) (Synthes, Paoli, PA). Two test groups were evaluated: 1. Fixation with DHS, and 2. Fixation with a DHS and calcium phosphate bone cement (Norian SRS (Skeletal Repair System)) augmentation of the fracture line and posteromedial calcar region of the proximal femur. Each femur was loaded to 1,650 N (2.5 body weight) for 10,000 cycles to simulate postoperative load transmission across the fracture construct during normal gait. The load was further increased successively by one body weight for another 10,000 cycles until failure. We evaluated fixation by measuring the amount of sliding of the lag screw of the DHS (shortening) and stiffness of the overall fracture construct (stability). SRS cement-augmented specimens had less shortening (1 mm versus 17 mm) and twice the initial construct stiffness compared to control specimens.


[Experience in treatment of Mycoplasma pneumoniae with acetylspiramycin].
Antibiotics of tetracycline and macrolide groups are mainly used in treatment of Mycoplasma pneumonia. In this study, acetylspiramycin (ASPM), an antibiotic of macrolide group, was given to 15 cases of Mycoplasma pneumoniae who visited this institute during the period from January, 1980 to March, 1981, and its clinical effects and side effects were investigated. The therapeutic effects were evaluated by days to the normal body temperature, to improvement in cough and to improvement in X-ray findings. The increase in serum antibody value was adopted as the diagnostic index of Mycoplasma infection. The patients who entered into this study were 7 males and 8 females, ranged from 19 to 60 years of age with an average of 36.3 years. The daily dose of ASPM was 600 to 1,200 mg (potency), and the mean administration period was 18.1 days. The results obtained were as follows. 1. The temperature fell to the normal within 2 to 7 days. 2. Cough disappeared in 2 days at shortest and in 55 days at longest. The mean period of cough disappearance was 14.5 days. 3. In 10 cases who could be followed up, shadows in X-ray films disappeared in 3 days at shortest and in 40 days at longest. The mean period to shadow disappearance was 18.6 days. 4. As regards clinical effects, marked improvement was obtained in 2 cases, improvement in 9, slight improvement in 4, and no change in 0 (improvement rate: 73.3%). 5. As a side effect, nausea was found in 1 case, but it was improved by discontinuance of administration.


Adenosine diphosphate ribose transferase from baby-hamster kidney cells (BHK-21/C13). Characterization of the reaction and product.
Some properties of ADP-ribose transferase, and its reaction product, from BHK-21/C13 cells are described. Enzyme activity was found almost exclusively in nuclei (90%), with the remaining 10% located in the cytosolic fraction. The nuclear enzyme is chromatin-bound and requires bivalent cations, preferably Mg2+, a pH of 8.0 and a temperature of 25 degrees C for optimal activity. Chromatin preparations incorporated radioactivity from [14C]NAD+ into acid-insoluble material for about 60 min. Kinetics for substrate NAD+ utilization were not of Michaelis--Menten type; biphasic kinetics were shown from a double-reciprocal plot (1/reaction velocity against 1/[NAD+]) and from a 'Hofstee' plot (reaction velocity/[NAD+] against reaction velocity). The transferase is unstable in the absence of Mg2+ ions. It is inhibited by thymidine, nicotinamide and nicotinamide analogues, but not by ATP, which stimulates it at concentrations of 5 mM and above. The enzyme requires thiol groups for activity; it is readily inhibited by N-ethylmaleimide at 0.5 mM. The product of the reaction is stable under acid conditions at temperatures up to 25 degrees C, but it is hydrolysed by HClO4 at 70 degrees C. It is resistant to NaOH, but is cleaved from its attachment to protein with alkali into trichloroacetic acid-insoluble and -soluble components. On the basis of Cs2SO4- density-gradient analysis under denaturing conditions (gradients included urea and guanidinium hydrochloride), and analysis of the reaction product directly on hydroxyapatite, we conclude that most of the radioactive ADP-ribose residues are firmly bound to protein, presumably in covalent linkage. Hydroxyapatite-chromatographic analysis of ADP-ribose residues released from protein by alkaline digestion showed a spectrum of molecular sizes including mono-, oligo- and poly-(ADP-ribose), when chromatin was incubated initially with [14C]NAD+ for 10 min and then for a further 30 min after addition of excess non-radioactive NAD+, only about 10% of the radioactive mono-(ADP-ribose) could be 'chased' into longer-chain molecules. Hydroxyapatite analysis was also used to show that, whereas all ADP-ribose residues were released from protein with NaOH, only 50% of them were susceptible to hydroxylamine. These hydroxylamine-sensitive residues included all size classes, although mono-(ADP-ribose) predominated. Finally, there was an approximately equal distribution of ADP-ribose incorporated into HCl-soluble proteins (including the histones) and HCl-insoluble proteins (including the non-histone proteins) when chromatin was incubated with NAD+ up to 0.5 mM, but at higher NAD+ concentrations more ADP-ribose was incorporated into the HCl-soluble fraction (82% at 4.0 mM-NAD+).


Effect of N-terminal and Met23 mutations on the structure and dynamics of onconase.
Onconase (rONC), otherwise known as ranpirnase or P-30 protein, which was initially purified from extracts of Rana pipiens oocytes and early embryos, exhibits anticancer activity both in vitro and in vivo and is in phase III clinical trials for tumor therapy. We have determined the solution NMR structure of a recombinant onconase with Met(-1), Gln1, and Leu23 residues (M-1, Q1, M23L)rONC. The 20 best solution structures had a backbone root mean square deviation of 0.41 +/- 0.09 A with respect to the average structure. The energy-minimized average NMR structure had a backbone root mean square deviation of 0.72 A from the x-ray crystallographic structure of native onconase; however, the orientation of the N-terminal residue in the two structures was very different. Comparison of the 15N HSQC spectrum of (M-1, Q1, M23L)rONC with that of a mutant E1S-rONC, which is identical to the nONC except with the N-terminal pyroglutamyl residue replaced by Ser, showed that N-terminal and residue 23 mutations induced structural changes in regions beyond the mutation sites. Model-free analysis of the backbone amide 15N-T1, 15N-T2, and 15N-1H NOE relaxation data for (M-1, Q1, M23L)rONC and E1S-rONC revealed that the E1S-rONC molecule showed very little flexibility, whereas (M-1, Q1, M23L)rONC exhibited substantial flexibility, which may account for the previously observed reduced stability and increased protease susceptibility. The alpha1 helix and beta-sheets of (M-1, Q1, M23L)rONC displayed bending motions. These data provided strong evidence for the presence of an N-terminal hydrogen bond network in E1S-rONC, but not in (M-1, Q1, M23L)rONC.


Thermal modification of connective tissues: basic science considerations and clinical implications.
Thermal modification (shrinkage) of capsular connective tissue has gained increasing popularity as an adjunctive or even a primary procedure in the arthroscopic treatment of shoulder instability. Although the physical effects of heat on collagenous tissues are well known, the long-term biologic fate of these shrunken tissues is still a matter of debate. The temperatures required to alter the molecular bonding of collagen and thus cause tissue shrinkage (65 degrees C to 70 degrees C) are also known to destroy cellular viability. Therefore, thermally modified tissues are devitalized and must undergo a biologic remodeling process. During this remodeling, the mechanical properties of the treated tissues are altered (decreased stiffness) and can be at risk for elongation if the postoperative rehabilitation regimen is too aggressive. Although anecdotal reports suggest that thermal capsular shrinkage does have a beneficial effect, the exact mechanism responsible for this clinical improvement has yet to be fully defined. The reported improvement could be due to the maintenance of initial capsular shrinkage, secondary fibroplasia and resultant thickening of the joint capsule, a loss of afferent sensory stimulation due to the destruction of sensory receptors, or a combination of all three. The clinical role for thermal modification of connective tissues has not yet been defined, but it appears that it may prove most useful as a stimulant for inducing a biologic repair response.


Archaeal extrachromosomal genetic elements.
Research on archaeal extrachromosomal genetic elements (ECEs) has progressed rapidly in the past decade. To date, over 60 archaeal viruses and 60 plasmids have been isolated. These archaeal viruses exhibit an exceptional diversity in morphology, with a wide array of shapes, such as spindles, rods, filaments, spheres, head-tails, bottles, and droplets, and some of these new viruses have been classified into one order, 10 families, and 16 genera. Investigation of model archaeal viruses has yielded important insights into mechanisms underlining various steps in the viral life cycle, including infection, DNA replication and transcription, and virion egression. Many of these mechanisms are unprecedented for any known bacterial or eukaryal viruses. Studies of plasmids isolated from different archaeal hosts have also revealed a striking diversity in gene content and innovation in replication strategies. Highly divergent replication proteins are identified in both viral and plasmid genomes. Genomic studies of archaeal ECEs have revealed a modular sequence structure in which modules of DNA sequence are exchangeable within, as well as among, plasmid families and probably also between viruses and plasmids. In particular, it has been suggested that ECE-host interactions have shaped the coevolution of ECEs and their archaeal hosts. Furthermore, archaeal hosts have developed defense systems, including the innate restriction-modification (R-M) system and the adaptive CRISPR (clustered regularly interspaced short palindromic repeats) system, to restrict invasive plasmids and viruses. Together, these interactions permit a delicate balance between ECEs and their hosts, which is vitally important for maintaining an innovative gene reservoir carried by ECEs. In conclusion, while research on archaeal ECEs has just started to unravel the molecular biology of these genetic entities and their interactions with archaeal hosts, it is expected to accelerate in the next decade.


Retrograde blood flow in the internal jugular veins of humans with hypertension may have implications for cerebral arterial blood flow.
To use multi-parametric magnetic resonance imaging (MRI) to test the hypothesis that hypertensives would have higher retrograde venous blood flow (RVBF) in the internal jugular veins (IJV) vs. normotensives, and that this would inversely correlate with arterial inflow and gray matter, white matter, and cerebrospinal fluid volumes. Following local institutional review board approval and written consent, a prospective observational 3-T MRI study of 42 hypertensive patients (53 ± 2 years, BMI 28.2 ± 0.6 kg/m2, ambulatory daytime systolic BP 148 ± 2 mmHg, ambulatory daytime diastolic BP 101 ± 2 mmHg) and 35 normotensive patients (48 ± 2 years, BMI 25.2 ± 0.8 kg/m2, ambulatory daytime systolic BP 119 ± 3 mmHg, ambulatory daytime diastolic BP 90 ± 2 mmHg) was performed. Phase contrast imaging calculated percentage retrograde venous blood flow (%RVBF), brain segmentation estimated regional brain volumes from 3D T1-weighted images, and pseudo-continuous arterial spin labeling measured regional cerebral blood perfusion. Statistical analysis included two-sample equal variance Student's T tests, two-way analysis of variance with Tukey's post hoc correction, and permutation-based two-group general linear modeling (p < 0.05). In the left IJV, %RVBF was higher in hypertensives (6.1 ± 1.5%) vs. normotensives (1.1 ± 0.3%, p = 0.003). In hypertensives, there was an inverse relationship of %RVBF (permutation-based general linear modeling) to cerebral blood flow in several brain regions, including the left occipital pole and the cerebellar vermis (p < 0.01). Percentage retrograde flow in the left IJV correlated inversely with the total matter volume (gray plus white matter volume) in hypertensives (r = - 0.49, p = 0.004). RVBF in the left IJV is greater in hypertensives vs. normotensives and is linked to regional hypoperfusion and brain total matter volume. • Hypertensive humans have higher retrograde cerebral venous blood flow, associated with regional brain hypoperfusion and lower tissue volume, compared with controls. • Cerebral retrograde venous blood flow may add further stress to already hypoperfused tissue in hypertensive patients. • The amount of retrograde venous blood flow in hypertensive patients may predict which patients might be at higher risk of developing cerebral pathologies.


Alcohol consumption and pancreatitis mortality in Russia.
Pancreatitis is a major public health problem with high associated economic costs. The incidence of pancreatitis has increased in many European countries in recent decade. Accumulated research and empirical evidence suggests that excessive alcohol consumption is a major risk factor for both acute and chronic pancreatitis. The aim of this study was to examine the aggregate-level relation between the alcohol consumption and pancreatitis mortality rates in Russia. Age-standardized sex-specific male and female pancreatitis mortality data for the period 1970-2005 and data on overall alcohol consumption were analyzed by means ARIMA (autoregressive integrated moving average) time series analysis. Alcohol consumption was significantly associated with both male and female pancreatitis mortality rates: a 1 liter increase in overall alcohol consumption would result in a 7.0% increase in the male pancreatitis mortality rate and in 2.3% increase in the female mortality rate. The results of the analysis suggest that 63.1% of all male pancreatitis deaths and 26.8% female deaths in Russia could be attributed to alcohol. Conclusions The outcomes of this study provide indirect support for the hypothesis that unfavorable mixture of higher overall level of alcohol consumption and binge drinking pattern is an important contributor to the pancreatitis mortality rate in Russian Federation.


Dendrotoxin-κ suppresses tumor growth induced by human lung adenocarcinoma A549 cells in nude mice.
Voltage-gated K(+) (Kv) channels have been considered to be a regulator of membrane potential and neuronal excitability. Recently, accumulated evidence has indicated that several Kv channel subtypes contribute to the control of cell proliferation in various types of cells and are worth noting as potential emerging molecular targets of cancer therapy. In the present study, we investigated the effects of the Kv1.1-specific blocker, dendrotoxin-κ (DTX-κ, on tumor formation induced by the human lung adenocarcinoma cell line A549 in a xenograft model. Kv1.1 mRNA and protein was expressed in A549 cells and the blockade of Kv1.1 by DTX-κ, reduced tumor formation in nude mice. Furthermore, treatment with DTX-κ significantly increased protein expression of p21(Waf1/Cip1), p27(Kip1), and p15(INK4B) and significantly decreased protein expression of cyclin D3 in tumor tissues compared to the control. These results suggest that DTX-κ has anti-tumor effects in A549 cells through the pathway governing G1-S transition.


Combining Heparin and a FX/Xa Aptamer to Reduce Thrombin Generation in Cardiopulmonary Bypass and COVID-19.
Known limitations of unfractionated heparin (UFH) have encouraged the evaluation of anticoagulant aptamers as alternatives to UFH in highly procoagulant settings such as cardiopulmonary bypass (CPB). Despite progress, these efforts have not been totally successful. We take a different approach and explore whether properties of an anticoagulant aptamer can complement UFH, rather than replace it, to address shortcomings with UFH use. Combining RNA aptamer 11F7t, which targets factor X/Xa, with UFH (or low molecular weight heparin) yields a significantly enhanced anticoagulant cocktail effective in normal and COVID-19 patient blood. This aptamer-UFH combination (1) supports continuous circulation of human blood through an ex vivo membrane oxygenation circuit, as is required for patients undergoing CPB and COVID-19 patients requiring extracorporeal membrane oxygenation, (2) allows for a reduced level of UFH to be employed, (3) more effectively limits thrombin generation compared to UFH alone, and (4) is rapidly reversed by the administration of protamine sulfate, the standard treatment for reversing UFH clinically following CPB. Thus, the combination of factor X/Xa aptamer and UFH has significantly improved anticoagulant properties compared to UFH alone and underscores the potential of RNA aptamers to improve medical management of acute care patients requiring potent yet rapidly reversible anticoagulation.


Threshold conditions for synaptically evoking Ca(2+) waves in hippocampal pyramidal neurons.
Regenerative Ca(2+) release from inositol 1,4,5-trisphosphate (IP(3))-sensitive intracellular stores in the form of Ca(2+) waves leads to large-amplitude [Ca(2+)](i) increases in the apical dendrites of hippocampal CA1 pyramidal neurons. Release is generated following synaptic activation of group I metabotropic glutamate (mGlu) receptors. We systematically examined the conditions for evoking these waves in transverse slices from 2- to 3-wk-old rats. Using a sharpened asymmetrical bipolar tungsten stimulating electrode placed in the stratum radiatum, we varied the lateral position of the electrode, the number of stimulating pulses, the train frequency, and stimulus current. Several trends were clear. Increasing the frequency of stimulation from 20 to 100 Hz, keeping the total number of pulses constant, lowered the required stimulus current. Stimulation at frequencies below 20 Hz made it difficult to evoke release. Increasing the number of stimulation pulses, keeping the frequency constant, lowered the threshold current. A minimum of five pulses at 100 Hz was required to evoke release reliably, but several examples of success with three pulses were recorded. Theta-burst stimulation was as effective as tetanic stimulation. Placing the point of the stimulation electrode closer to the pyramidal neuron made it easier to evoke release, although stimulation at a lateral distance of 500 microm with unsharpened electrodes was sometimes successful. The simplest explanation for these results is that a bolus of IP(3) must be produced quickly in a restricted region of the dendrites to generate Ca(2+) waves. The conditions necessary for evoking regenerative Ca(2+) release have many parallels (and some differences) with the conditions required to evoke long-term potentiation in these cells following tetanic stimulation.


The Lived Experiences of Iranian Patients with Rheumatoid Arthritis: A Qualitative Research.
Rheumatoid Arthritis (RA) is a chronic inflammatory disease that affects various systems of the body. The present study was conducted with the aim to explain the lived experiences of Iranian patients with RA. According to the research question, the phenomenological research approach was used. The participants were selected through purposive sampling, and sampling was continued until data saturation was achieved. The data was collected by conducting unstructured interviews and using an audio recorder. To interpret the data, an interpretive/hermeneutic approach was implemented using Van Manen's method. In addition, Lincoln and Guba's criteria were used to evaluate data robustness. A total of 24 participants participated in the study. Overall, 145 basic concept codes were extracted which were classified into the four main themes of self-management with the participation of the family, spiritual resilience in the face of existing problems, the tendency to hide the disease, and the fear of an uncertain future. It is recommended that care providers use the findings of this study to develop care plans in the various aspects of the physical, mental, social, and spiritual needs of patients with RA based on their concerns and deep experiences. It is also suggested that other qualitative research approach methodologies such as grounded theory for explaining the process of patient care and providing an appropriate model, or action research for solving the problems of hospitalized patients through suitable care at home be implemented.


Reversible redox modifications in the microglial proteome challenged by beta amyloid.
Microglia are resident macrophages in the central nervous system, whose participation against exogenous injuries and infections is mainly marked by an immediate release of inflammatory cytokines along with a toxic efflux of superoxide radicals. Indeed, many lines of evidence indicate that persistent activation of these cells turns their neuroprotective phenotype into a neurotoxic one, which contributes to destroy neuronal activity and induces neuronal loss in several neurodegeneration processes, such as Alzheimer's disease. In this study we attempted to fill-in the gap in our knowledge about redox regulation of amyloid activated microglia. With this aim, we carried out a robust and comprehensive characterization of the reversibly redox modified proteome both at the level of resting and amyloid-activated BV2 cells, an immortalised cell line of murine microglia. The approach we used combined the selective enrichment of reversible redox modified proteins through a biotin bait with nanoscale liquid chromatography tandem mass spectrometry of their proteolytic peptides. By this reliable approach, we identified 60 proteins changing the redox status of their selective cysteine residues upon treatment with the amyloidogenic Aβ25-35 peptide. These results assessed that in microglia stimulated by amyloids, redox modifications of the proteome specifically target proteins involved in crucial cell processes, i.e. those involved in the protein synthesis. In particular, for peroxiredoxin-6 (Prdx6) and Ras-related C3 botulinum toxin substrate 1 (Rac1) we suggest mechanisms through which reversible redox modifications could affect the peculiar role of microglia in amyloidogenic injury, which at the same time reinforce the oxidative burst and resist toward it. Moreover, the redox modulation we observed on chloride intracellular channel protein 1 (CLIC1) strengthens the structural and functional relationship between the oxidative stress and the metamorphic transition of this protein from a soluble form to an integral membrane form. The redox signatures we determined might also provide neurologists with more specific and reliable biomarkers to distinguish the diverse microglia status in neurodegeneration and then to drive targeted drug design.


Gut Microbiota Modulate CD8 T Cell Responses to Influence Colitis-Associated Tumorigenesis.
There is increasing evidence that gut microbiome perturbations, also known as dysbiosis, can influence colorectal cancer development. To understand the mechanisms by which the gut microbiome modulates cancer susceptibility, we examine two wild-type mouse colonies with distinct gut microbial communities that develop significantly different tumor numbers using a mouse model of inflammation-associated tumorigenesis. We demonstrate that adaptive immune cells contribute to the different tumor susceptibilities associated with the two microbial communities. Mice that develop more tumors have increased colon lamina propria CD8+ IFNγ+ T cells before tumorigenesis but reduced CD8+ IFNγ+ T cells in tumors and adjacent tissues compared with mice that develop fewer tumors. Notably, intratumoral T cells in mice that develop more tumors exhibit increased exhaustion. Thus, these studies suggest that microbial dysbiosis can contribute to colon tumor susceptibility by hyperstimulating CD8 T cells to promote chronic inflammation and early T cell exhaustion, which can reduce anti-tumor immunity.


Fimbrolide-coated antimicrobial lenses: their in vitro and in vivo effects.
To examine the ability of contact lenses coated with fimbrolides, inhibitors of bacterial quorum sensing, to prevent microbial adhesion and their safety during short-term clinical assessment. A fimbrolide was covalently attached to commercially available high Dk contact lenses. Subsequently Pseudomonas aeruginosa, Staphylococcus aureus, Serratia marcescens, or Acanthamoeba sp. were added to the lenses and control uncoated contact lenses. Lenses plus microbes were incubated for 24 h, then washed thoroughly to remove non-adherent microbes. Lenses were macerated and resulting slurry plated onto agar plates. After appropriate incubation, the numbers of colony forming units of bacteria (or numbers of Acanthamoeba trophozoites measured using a hemocytometer) from fimbrolide-coated and uncoated lenses were examined. A Guinea Pig model of lens wear was used to assess the safety of lenses worn on a continuous basis for 1 month. In a separate study, 10 subjects wore fimbrolide-coated lenses for 24 h. The responses of the Guinea Pigs and human volunteers to the lenses were assessed by slit lamp examination. The fimbrolides-coated lenses reduced the adhesion of all bacterial strains tested, with reductions occurring of between 67 and 92%. For Acanthamoeba a reduction of 70% was seen. There were no significant differences in ocular responses to fimbrolide-coated lenses compared with controls in either the 1 month animal model or overnight human trial. Fimbrolide-coated lenses show promise as an antibacterial and anti-acanthamoebal coating on contact lenses and appear to be safe when worn on the eye in an animal model.


Evolution of a hereditary cardiac conduction and muscle disorder: a study involving a family with six generations affected.
This study describes six generations of a family with autosomal dominant cardiac conduction system and myocardial disease with recognizable clinical stages. A 20 year follow-up of nine family members, a medical questionnaire of 196, electrocardiographic screening of 91, noninvasive testing of 20, and catheterization with endomyocardial biopsy of six are the basis of this report. The clinical stages are as follows: Stage I occurs in the second and third decades of life and is characterized by an absence of symptoms, normal heart size, sinus bradycardia, and premature atrial contractions. Stage II is marked by first-degree atrioventricular block in the third and fourth decades. Stage III occurs in the fourth and fifth decades and is accompanied by chest pain, fatigue, lightheadedness, and advanced atrioventricular block followed by the development of atrial fibrillation or flutter. Stage IV, in the fifth and sixth decades of life, is characterized by congestive heart failure and recurrent ventricular arrhythmias. Light microscopy of right ventricular endomyocardial biopsy specimens from patients in stage II revealed very mild fibrosis; electron microscopy of the specimens demonstrated mild dilatation of tubules, mitochondrial swelling, and minimal myofibrillar loss. Biopsy specimens from patients with stage III disease were similar to those from patients with stage II disease except for an increase of myofibrillar loss. The stage IV specimens had diffuse fibrosis and more severe tubular dilatation, mitochondrial cristolysis, and myofibrillar loss. At autopsy in the proband, the atrial changes were more severe than the ventricular and were especially marked in the sinoatrial and atrial myocardium. Early recognition of the disease and use of pacemakers and antiarrhythmic agents have proved beneficial for affected family members. Thorough family studies of patients with conduction system disease and/or dilated cardiomyopathy are necessary to better understand the hereditary basis and natural course of this category of disease.


The equiprobability bias from a mathematical and psychological perspective.
The equiprobability bias (EB) is a tendency to believe that every process in which randomness is involved corresponds to a fair distribution, with equal probabilities for any possible outcome. The EB is known to affect both children and adults, and to increase with probability education. Because it results in probability errors resistant to pedagogical interventions, it has been described as a deep misconception about randomness: the erroneous belief that randomness implies uniformity. In the present paper, we show that the EB is actually not the result of a conceptual error about the definition of randomness. On the contrary, the mathematical theory of randomness does imply uniformity. However, the EB is still a bias, because people tend to assume uniformity even in the case of events that are not random. The pervasiveness of the EB reveals a paradox: The combination of random processes is not necessarily random. The link between the EB and this paradox is discussed, and suggestions are made regarding educational design to overcome difficulties encountered by students as a consequence of the EB.


Interaction of mammalian O(6)-alkylguanine-DNA alkyltransferases with O(6)-benzylguanine.
Human O(6)-alkylguanine-DNA alkyltransferase (hAGT) activity is a major factor in providing resistance to cancer chemotherapeutic alkylating agents. Inactivation of hAGT by O(6)-benzylguanine (BG) is a promising strategy for overcoming this resistance. Previous studies, which have focused on the region encompassed by residues Pro138 to Gly173, have identified more than 100 individual mutations located at 23 discrete sites at which alterations can render AGT less sensitive to BG. We have now extended the examination of possible sites in hAGT at which alterations might lead to BG resistance to include the residues from Val130 to Asn137, which also make up part of the binding pocket into which BG is postulated to fit. A further 21 mutations located at positions Gly132, Met134, Arg135, and Gly136 were found to lower sensitivity to BG. Mutants R135L, R135Y, and G136P were the most strikingly resistant, with a 50-fold increase in the amount of BG needed to obtain 50% inactivation. These results therefore increase the number of sites at which BG resistance can occur in response to a single amino acid change to 27. Although mammalian AGTs are very similar in amino acid sequence, mouse AGT (mAGT) is significantly less sensitive to BG than rat AGT (rAGT) or hAGT. Construction of chimeric proteins in which portions came from the rAGT and the mAGT indicated that the difference in inactivation resided solely in the amino acids located in the sequence from residues 150 to 188. Individual mutations of the three residues where rAGT and mAGT differ in this region showed that the principal reason for the reduced ability of the mAGT to react with BG was the presence of a histidine residue at position 161, which is occupied by asparagine in rAGT and hAGT. These experiments indicate that many minor changes in amino acids forming all parts of the nucleoside binding pocket of AGT can alter its ability to react with BG and that the possibility that polymorphisms or variants may occur reducing the effectiveness of combination therapy with BG and alkylating agents must be considered.


Association between prenatal maternal infection and disordered eating behaviours in adolescence: a UK population-based prospective birth cohort study.
Prenatal infections have been proposed as a putative risk factor for a number of psychiatric outcomes across a continuum of severity. Evidence on eating disorders is scarce. We investigated whether exposure to prenatal maternal infections is associated with an increased risk of disordered eating and weight and shape concerns in adolescence in a large UK birth cohort. We used data from the Avon Longitudinal Study of Parents and Children. The primary exposure was maternal experience of infections at any time in pregnancy. Study outcomes were presence of any, monthly or weekly disordered eating at 14 and 16 years of age, and weight and shape concerns at 14 years. We defined the causal effect of the exposure on these outcomes using a counterfactual framework adjusting our analyses for a number of hypothesised confounders, and imputing missing confounder data using multiple imputation. In total, 4884 children had complete exposure and outcome data at age 14 years, and 4124 at 16 years. Exposed children had a greater risk of reporting weekly disordered eating at both age 14 [risk difference (RD) 0.9%, 95% confidence interval (CI) -0.01 to 1.9, p = 0.08] and 16 (RD 2.3%, 95% CI 0.6-3.9, p < 0.01), though evidence of an association was weak at age 14 years. Exposed children also had greater weight and shape concerns at age 14 years (mean difference 0.15, 95% CI 0.05-0.26, p < 0.01). Exposure to prenatal maternal infection is associated with greater risk of disordered eating in adolescence. This association could be explained by in utero processes leading to impaired neurodevelopment or altered immunological profiles. Residual confounding cannot be excluded.


Description of Anaerobacterium chartisolvens gen. nov., sp. nov., an obligately anaerobic bacterium from Clostridium rRNA cluster III isolated from soil of a Japanese rice field, and reclassification of Bacteroides cellulosolvens Murray et al. 1984 as Pseudobacteroides cellulosolvens gen. nov., comb. nov.
An obligately anaerobic bacterial strain designated T-1-35(T) was isolated as a dominant cultivable cellulose-degrading bacterium from soil of a Japanese rice field as an anaerobic filter-paper degrader. Cells of strain T-1-35(T) stained Gram-positive and were non-spore-forming rods with rounded ends, 0.8-1.0×3.5-15.0 µm, and motile by means of two to four polar flagella. Cells of strain T-1-35(T) exhibited pleomorphism: in aged cultures (over 90 days of incubation), almost all cells were irregularly shaped. Although no spore formation was observed, cells tolerated high temperatures, up to 90 °C for 10 min. The temperature range for growth was 15-40 °C, with an optimum at 35 °C. The pH range for growth was 5.5-9.0, with an optimum at pH 8.0-8.5 (slightly alkaliphilic). Strain T-1-35(T) fermented some carbohydrates to produce ethanol and lactate as the major products. Major cellular fatty acids were iso-C16 : 0 and iso-C13 : 0 3-OH. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain T-1-35(T) belonged to Clostridium rRNA cluster III. The closest relative of strain T-1-35(T) was Bacteroides cellulosolvens WM2(T), with 16S rRNA gene sequence similarity of 93.4 %. Phenotypic, physiological and molecular genetic methods demonstrated that strain T-1-35(T) was distinct from its phylogenetic relatives (members of Clostridium rRNA cluster III) because it predominantly produced ethanol, iso-C13 : 0 3-OH was a major cellular fatty acid and it always exhibited pleomorphism. On the basis of the results of a polyphasic taxonomic study, strain T-1-35(T) is considered to represent a novel genus and species, Anaerobacterium chartisolvens gen. nov., sp. nov. The type strain of Anaerobacterium chartisolvens is T-1-35(T) ( = DSM 27016(T) = NBRC 109520(T)). In addition, from the results of our phylogenetic analysis and its phenotypic features, the species Bacteroides cellulosolvens Murray et al. 1984 is proposed to be reclassified in the new genus Pseudobacteroides as Pseudobacteroides cellulosolvens gen. nov., comb. nov., with the type strain WM2(T) ( = ATCC 35603(T) = DSM 2933(T) = NRCC 2944(T)).


Human lymphocytes: similarity of B and T cell surface morphology.
When viewed by scanning electron microscopy human lymphocytes fixed in suspension and processed with minimal cell loss appear uniformly covered with short microvilli. Contrary to previous reports, lymphocytes from subpopulations selectively enriched for T cells are villous and indistinguishable from B lymphocytes. Whereas lymphocyte surface architecture can change rapidly and substantially in response to environmental modifications, such as contact with an underlying surface, these alterations are similar for both B and T cells and do not serve to distinguish these subpopulations.


Experience of methadone therapy in 100 consecutive chronic pain patients in a multidisciplinary pain center.
The objective of the study was to describe the experience of methadone use in 100 consecutive chronic pain patients managed in a single multidisciplinary center. A chart review of chronic pain patients on methadone therapy initiated at the Wasser Pain Management Center from January 2001 to June 2004. SETTING, PATIENTS, AND INTERVENTION: Outpatients receiving methadone for chronic pain management in a tertiary multidisciplinary pain center. Effects on pain relief and function, conversion ratio from other opioids, side effects, and disposition were reviewed. Charts of 100 methadone patients (age 45 +/- 11 years old; M/F: 3/7; duration of pain 129 +/- 110 months) managed by five physicians and one nurse were reviewed. The main reason for the initiation of methadone therapy was opioid rotation (72%). The average oral morphine equivalent dose was 77 mg/day before methadone therapy, and the methadone dose after initial stabilization was 42 mg with no consistent conversion ratio observed. The mean duration of methadone therapy was 11 months. Most of the patients (91%) were taking concomitant adjuvant analgesics or psychotropic agents, mostly antidepressants and anticonvulsants. The average Numeric Verbal Rating Score before and after methadone treatment was 7.2 +/- 1.7 and 5.2 +/- 2.5 (P < 0.0001). Thirty-five patients discontinued their methadone treatment mainly because of side effects, ineffectiveness, or both. From our experience, methadone is an effective alternative to conventional opioids for chronic pain management when used by experienced clinicians in a setting that allows for close monitoring and careful dose initiation and adjustment.


Adenomyosis: Disease, uterine aging process leading to symptoms, or both?
For many decades adenomyosis has been a histological diagnosis in hysterectomy specimens. Traditionally, it has been considered a disease of late reproductive and premenopausal years causing uterine enlargement, dysmenorrhoea and menorrhagia. Recent advances in pelvic and uterine imaging techniques including transvaginal sonography and magnetic resonance imaging were responsible for a shift towards a non-invasive diagnosis and made a significant contribution to a better understanding of its pathogenesis, epidemiology, histological spectrum, and clinical symptomatology. With these non-invasive tools it has been shown that adenomyosis is probably a condition affecting much younger populations and is frequently asymptomatic at an early stage of its development. Regarding symptomatic disease, the distribution and extent of adenomyotic lesions do not correlate consistently with the various symptoms that are considered typical of adenomyosis. More importantly, accurate diagnosis of adenomyosis suffers from a lack of consensus among experts on imaging and even histological diagnostic criteria. Several pathogenetic theories have attempted to shed light on the establishment, evolution and distribution of adenomyotic lesions within the uterine wall, including the tissue injury and repair (TIAR) mechanism, metaplasia, and the more recent genetic-epigenetic theory. So far, none of these can adequately and independently explain the appearance of all types of adenomyosis. This review paper attempts a correlation between the proposed pathogenetic theories and the clinical and histological spectrum of adenomyosis, in an effort to give a plausible explanation of the evolution of this condition from an asymptomatic state to a disease, through synthesis of the existing data.


Effects of parasitic sex-ratio distorters on host genetic structure in the Armadillidium vulgare-Wolbachia association.
In the pill bug Armadillidium vulgare (Crustacea, Oniscidea), Wolbachia facilitates its spread through vertical transmission via the eggs by inducing feminization of genetic males. The spread of feminizing Wolbachia within and across populations is therefore expected to influence mitochondrial DNA (mtDNA) genetic structure by hitchhiking. To test this hypothesis, we analysed nuclear and mtDNA genetic structure, and Wolbachia prevalence in 13 populations of the pill bug host. Wolbachia prevalence (ranging from 0% to 100% of sampled females) was highly variable among populations. All three Wolbachia strains previously observed in A. vulgare were present (wVulC, wVulM and wVulP) with wVulC being the most prevalent (nine of 13 populations). The host showed a genetic structure on five microsatellite loci that is compatible with isolation by distance. The strong genetic structure observed on host mtDNA was correlated with Wolbachia prevalence: three mitotypes were in strong linkage disequilibrium with the three strains of Wolbachia. Neutrality tests showed that the mtDNA polymorphism is not neutral, and we thus suggest that this unusual pattern of mtDNA polymorphism found in A. vulgare was due to Wolbachia.


Rationale and design of the randomized, multicenter, cilostazol for RESTenosis (CREST) trial.
Restenosis of a segment of diseased coronary artery following metallic stenting is a common clinical problem and a major limitation of the procedure. Systemic pharmacologic interventions to deal with this problem have met with little success. Several small studies suggest that cilostazol, a phosphodiesterase III inhibitor whose pharmacologic properties include antiplatelet, antithrombotic, and vasodilatory effects; a beneficial effect on serum lipids; and in vitro inhibition of smooth muscle cell proliferation, may help prevent platelet aggregation and impede the accumulation of new intimal tissue in the stented artery. The Cilostazol for RESTenosis (CREST) trial will aim to evaluate more definitively the ability of cilostazol to prevent restenosis following uncomplicated stent implantation for de novo coronary artery stenosis. In this randomized, double-blind, multicenter study, 700 patients will receive clopidogrel, aspirin, and either cilostazol or placebo after successful intracoronary stent implantation. The primary endpoint is minimal luminal diameter (MLD) of the first lesion stented after 6 months; secondary endpoints include MLD in all lesions, mean percent diameter stenosis, target lesion revascularization, and major angiographic endpoints. Safety endpoints are abnormal complete blood count and liver function tests at 1, 3, and 6 months. The trial has been initiated, and enrollment is anticipated to be concluded in 2003. Cilostazol has properties that may reduce or avert in-stent coronary restenosis. The CREST trial is a large, rigorously conducted trial that may corroborate the favorable effects of cilostazol on coronary stent restenosis suggested by earlier studies.


Osteogenic efficiency of demineralized and lyophylized xenogeneic bone and syngeneic dentine implants in mice.
Intramuscular implantation of demineralized and lyophilized rat bone matrix and murine lower incisors into thigh muscles of BALB/c mice results in deposits of bone adjacent to the implants, a phenomenon termed as ectopic osteogenesis. The yield of induced bone does not critically depend on the mass of implanted matrices, and thus on the quantity of bone morphogenetic proteins (BMPs) present in the implants. A positive correlation between bone matrix implant weight and the yield of induced bone was observed only 28 days post grafting, i.e. when endochondreal osteogenesis is completed and bone resorption has not advanced. A more consistent yield of bone induction wasobserved in the case of demineralized tooth implants. It is postulated that chondro/osteoinduction by demineralized, lyophilized matrix implants is not determined by the range of BMPs presumably released in proportion to implant size, but is rather limited by the population of responsive host mesenchymal cells.


A novel interferometric method for the study of the viscoelastic properties of ultra-thin polymer films determined from nanobubble inflation.
Glass formation and glassy behavior remain as the important areas of investigation in soft matter physics with many aspects which are still not completely understood, especially at the nanometer size-scale. In the present work, we show an extension of the "nanobubble inflation" method developed by O'Connell and McKenna [Rev. Sci. Instrum. 78, 013901 (2007)] which uses an interferometric method to measure the topography of a large array of 5 μm sized nanometer thick films subjected to constant inflation pressures during which the bubbles grow or creep with time. The interferometric method offers the possibility of making measurements on multiple bubbles at once as well as having the advantage over the AFM methods of O'Connell and McKenna of being a true non-contact method. Here we demonstrate the method using ultra-thin films of both poly(vinyl acetate) (PVAc) and polystyrene (PS) and discuss the capabilities of the method relative to the AFM method, its advantages and disadvantages. Furthermore we show that the results from experiments on PVAc are consistent with the prior work on PVAc, while high stress results with PS show signs of a new non-linear response regime that may be related to the plasticity of the ultra-thin film.


Isolation and characterization of a novel liver-derived immunoinhibitory factor.
Cytosolic extracts prepared from perfused whole liver or purified hepatocytes of C57BL/6 mice inhibited interleukin-2--and concanavalin A--induced spleen cell proliferation in vitro. In contrast, cytosolic extracts from purified nonparenchymal liver cells had no effect. Arginase and very-low-density lipoprotein were previously identified as two immuninhibitory substances present in liver cytosolic extracts. We demonstrated, however, that inhibitory activity remained after removal of very-low-density lipoprotein and arginase from liver cytosolic extract by repeated ultracentrifugation and gel filtration chromatography, respectively, suggesting the presence of another inhibitor. Further purification by anion-exchange chromatography and chromatofocusing led to the isolation of a novel liver-derived immunohibitory factor. This liver-derived immunoinhibitory factor is sensitive to pronase digestion and heat and acid treatment; it has an estimated isoelectric point of 8.25. The Mr of liver-derived immunoinhibitory factor is 28 kD as estimated from its migration on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which is identical under both reducing and nonreducing conditions, indicating a monomeric nature of this protein. Amino acid composition analysis discloses that liver-derived immunoinhibitory factor is relatively rich in glycine and proline residues. Interleukin-2--induced spleen cell proliferation in vitro is inhibited by ths liver-derived immunoinhibitory factor, with a 50% inhibitory dose of 1.4 nmol/L. Furthermore, the biological activity of the liver-derived immunoinhibitory factor is not confined to mouse spleen cells, since the growth of B16 mouse melanoma and H35 rat hepatoma cells is also inhibited. A comparison with other liver-derived immunoinhibitors reported previously supports our claim that the liver-derived immunoinhibitory factor is a novel inhibitory protein.


A phase II randomized trial of RAdium-223 dichloride and SABR Versus SABR for oligomEtastatic prostate caNcerS (RAVENS).
Metastasis directed therapy (MDT) for patients with oligometastatic disease is associated with improvements in progression free survival (PFS) and overall survival (OS) compared to systemic therapy alone. Additionally, within a prostate-cancer-specific cohort, MDT is able to forestall initiation of androgen deprivation therapy (ADT) in men with hormone-sensitive, oligometastatic prostate cancer (HSOPCa) compared to observation. While MDT appears to be safe and effective in HSOPCa, a large percentage of men will eventually have disease recurrence. Patterns of failure in HSOPCa demonstrate patients tend to have recurrence in the bone following MDT, raising the question of sub-clinically-apparent osseous disease. Radium-223 dichloride is a radiopharmaceutical with structural similarity to calcium, allowing it to be taken up by bone where it emits alpha particles, and therefore might have utility in the treatment of micrometastatic osseous disease. Therefore, the primary goal of the phase II RAVENS trial is to evaluate the efficacy of MDT + radium-223 dichloride in prolonging progression free survival in men with HSOPCa. Patients with HSOPCa and 3 or less metastases with at least 1 bone metastasis will be randomized 1:1 to stereotactic ablative radiation (SABR, also known as stereotactic body radiation therapy (SBRT)) alone vs SABR + radium-223 dichloride with a minimization algorithm to balance assignment by institution, primary intervention, prior hormonal therapy, and PSA doubling time. SABR is delivered in one to five fractions and patients in the SABR + radium-223 dichloride arm will receive six infusions of radium-223 dichloride at four-week intervals. The primary end point is progression free survival. The secondary clinical endpoints include toxicity and quality of life assessments, local control at 12 months, locoregional progression, time to distant progression, time to new metastasis, and duration of response. The RAVENS trial will be the first described phase II, non-blinded, randomized study to compare SABR +/- radium-223 dichloride in patients with HSOPCa and 3 or less metastases with at least one bone metastasis. The primary hypothesis is that SABR + radium-223 dichloride will increase median progression-free survival from 10 months in the SABR arm to 20 months in the SABR + radium-223 dichloride arm. Clinicaltrials.gov. Identifier: NCT04037358. Date of Registration: July 30, 2019. Date of First Participant Enrolled: August 9, 2019. Date of Last Approved Amendment: October 16, 2019. Protocol Version: Version 5.


Comparisons of overground endoscopy and treadmill endoscopy in UK Thoroughbred racehorses.
To date there is no information on the comparison of the more recently documented technique of performing endoscopy during ridden exercise in the field, with the more traditional method of endoscopy during high-speed treadmill exercise. To compare the results of upper respiratory tract endoscopy in UK Thoroughbred racehorses performed during ridden exercise in the field with those obtained during exercise on the treadmill. A direct comparison was undertaken in 4 horses whereby both procedures were performed in the same horse within 10 days of each other. An indirect comparison was also undertaken whereby the results of overground endoscopy performed in 50 racehorses was compared to the results obtained during treadmill endoscopy in a further 50 racehorses. Horses were matched for age, gender, use (National Hunt vs. Flat) and presenting complaint (abnormal respiratory noise vs. poor performance). Dorsal displacement of the soft palate was diagnosed less frequently during overground endoscopy than during treadmill endoscopy. There was no significant difference in the diagnosis of dynamic laryngeal collapse between the 2 techniques. The treadmill exercise test was performed over longer distances at higher inclines, albeit at lower speeds than the overground test. In contrast to the treadmill test, the overground test was frequently performed in intervals. The results of both the direct and indirect comparisons suggest that dorsal displacement of the soft palate is diagnosed less often during overground endoscopy than during treadmill endoscopy. Strenuous exercise tests may be more easily performed on a treadmill than by performing multiple exercise intervals in the field. Care should be taken in interpreting negative findings during both procedures, but particularly during overground endoscopy if racing conditions have not been appropriately replicated.


Biomarkers and echocardiographic predictors of myocardial dysfunction in patients with hypertension.
The study aimed to identify early echocardiographic and circulating biomarkers of heart failure (HF) in hypertensive patients with normal resting echocardiography. Echocardiography at rest and during exercise, and selected biomarkers were assessed in control group, dyspnea group, and HF group. On exercise dyspnea patients had lower early diastolic (E') and systolic (S') mitral annular velocity (12.8 ± 1.0 vs 14.9 ± 3.0 cm/sec and 9.3 ± 2.0 vs 10.9 ± 2.0 cm/sec, respectively), and higher E/E' ratio compared to control group (6.7 ± 1.0 vs 5.9 ± 1.0) (p < 0.05 for all comparisons). The level of N-terminal propeptide of procollagen type III (PIIINP) was significantly higher in dyspnea group than in controls (p = 0.01). Control and dyspnea patients had lower levels of cardiotrophin-1, cystatin C, syndecan-4, and N terminal-probrain natriuretic peptide than HF patients (all p ≤ 0.01). In multivariate analysis PIIINP (unadjusted odds ratio [OR] = 8.2, 95% confidence interval [Cl] 1.7-40.6; p = 0.001; adjusted OR = 8.7; 95%CI: 1.5-48.3; p = 0.001) and E/E' ratio on exercise (unadjusted OR = 1.8, 95%CI: 0.8-4.0; p = 0.033; adjusted OR = 2.0; 95%CI: 0.8-4.8; p = 0.012) were the only factors significantly associated with the presence of dyspnea. PIIINP is the first early biomarker for the HF development in patients with HA and normal resting echocardiography. Exertional echocardiography may indicate patients with incipient HF with preserved ejection fraction.


Tooth Aspiration in a Patient with Traumatic Brain Injury.
Tooth aspiration is a rare issue in the course of patients with trauma and may remain undiagnosed for a long period, resulting in delayed complications, such as atelectasis and recurrent infections. Flexible bronchoscopy is considered the preferred primary procedure for the management of airway foreign bodies in adults. However, it may cause intracranial hypertension in trauma patients with concomitant head injuries. We herein report a case of a patient with traumatic brain injury who underwent tooth aspiration using flexible bronchoscopy, with continuous monitoring of intracranial pressure (ICP). The importance of a thorough review of radiographs and chest computed tomography for foreign body aspiration in trauma patients was highlighted, particularly in a maxillofacial trauma, as tooth aspiration may remain undiagnosed for extended periods. Moreover, the difficulty in maintaining the ICP within normal limits during bronchoscopy in patients with traumatic brain injury was reinstated, and the need for continuous monitoring of the cerebral hemodynamics and harmonization was emphasized, with recommendations for bronchoscopy via an endotracheal tube.


Characterization of DNA polymerase beta mutants with amino acid substitutions located in the C-terminal portion of the enzyme.
We used quantitative complementation assays to characterize individual DNA polymerase beta (Pol beta) mutants for their ability to function in DNA replication and DNA repair. We also describe a screen for detecting mutator activity of DNA polymerase beta mutants. By using these bioassays, together with DNA polymerase activity gels, we characterized 15 new DNA polymerase beta mutants that display a wide spectrum of phenotypes. Most of these mutants are generally defective in their ability to synthesize DNA. However, two of our Pol beta mutants show more complex phenotypes: they are able to function in DNA repair but unable to participate in DNA replication. One of our mutants displays mutator activity in vivo. Our work provides a model to study mutant mammalian enzymes in Escherichia coli with phenotypes that are otherwise difficult to assess.


Fructose-Containing Plant Carbohydrates: Biological Activities and Medical Applications.
The review considers the chemical structure specifics and distribution in plants for fructose-containing carbohydrates (fructans). Various biological activities were observed in fructans and associated with their physicochemical features. Fructans affect many physiological and biochemical processes in the human body, improving health and reducing the risk of various disorders. Prebiotic activity is the most important physiological function of fructans. Fructans improve the microflora composition in the colon and intestinal mucosa by increasing the content of useful bacteria and decreasing the content of potentially harmful microorganisms, stimulate the physiological functions of the microflora, and provide for a better state of the intestine and a better health status. By modifying the intestinal microbiota and utilizing certain additional mechanisms, fructans can favorably affect the immune function, decrease the risk of various inflammatory processes, and to reduce the likelihood of tumorigenesis due to exposure to carcinogens. Fructans improve carbohydrate and lipid metabolism by reducing the blood levels of glucose, total cholesterol, low-density lipoprotein (LDL), and very-low-density lipoprotein (VLDL) and increasing the blood content of high-density lipoprotein (HLD). Fructans are low in calories, and their use in foods reduces the risk of obesity. Fructans facilitate higher calcium absorption and increase the bone density, thus reducing the risk of osteoporosis. Fructants protect the body from oxidative stress, intestinal infections, and parasitic invasions.


Chromium (VI) induced phytotoxicity and oxidative stress in pea (Pisum sativum L.): biochemical changes and translocation of essential nutrients.
Due to widespread industrial use, chromium (Cr) is considered a hazardous environmental pollutant. It is known to inhibit plant growth and development. The present study provides the evidence of the phytotoxicity of this metal on the pea (Pisum sativum L. cv Azad) plants. The plants of pea (Pisum sativum L.) were grown in refined sand under different concentrations i.e. 0.05, 0.1, 0.2, 0.3 and 0.4 mM of Cr (VI) in order to study the effect on growth and yield, photosynthetic pigments, relative water content, non-reducing sugar and protein with activity of certain enzymes like catalase, peroxidase, starch phosphorylase and ribonuclease. The analysis of the results showed that photosynthetic pigments (68.68%), relative water contents (62.77%), non-reducing sugar (66.66%) and protein (81.57%) were decrease along with reduction in plant height (52.69%) and leaf area (50.81%) of the pea plants. However, in response to various concentration of Cr exposed plants showed significant induction of reducing and total sugars with enzymes like catalase, starch phosphorylase and ribonuclease. The translocation of Cr in various part of pea plant have been found in order of root> stem> leaves>seeds which ranged between 34.8 to 217.3 mg g(-1) d.wt. (dry weight) in roots, 6.5 to 173.13 mg g(-1) d.wt. in shoot, 4.2 to 74.43 mg g(-1) d.wt. in leaves and 0.94 to 8.64 mg g(-1) d.wt. in seeds, that is also reflected by the transfer factor of Cr from refined sand to tested species.


Response of net primary production to land use and climate changes in the middle-reaches of the Heihe River Basin.
Net primary production (NPP) supplies matter, energy, and services to facilitate the sustainable development of human society and ecosystem. The response mechanism of NPP to land use and climate changes is essential for food security and biodiversity conservation but lacks a comprehensive understanding, especially in arid and semi-arid regions. To this end, taking the middle-reaches of the Heihe River Basin (MHRB) as an example, we uncovered the NPP responses to land use and climate changes by integrating multisource data (e.g., MOD17A3 NPP, land use, temperature, and precipitation) and multiple methods. The results showed that (a) land use intensity (LUI) increased, and climate warming and wetting promoted NPP. From 2000 to 2014, the LUI, temperature, and precipitation of MHRB increased by 1.46, 0.58°C, and 15.76 mm, respectively, resulting in an increase of 14.62 gC/m2 in annual average NPP. (b) The conversion of low-yield cropland to forest and grassland increased NPP. Although the widespread conversion of unused land and grassland to cropland boosted both LUI and NPP, it was not conducive to ecosystem sustainability and stability due to huge water consumption and human-appropriated NPP. Urban sprawl occupied cropland, forest, and grassland and reduced NPP. (c) Increase in temperature and precipitation generally improved NPP. The temperature decreasing <1.2°C also promoted the NPP of hardy vegetation due to the simultaneous precipitation increase. However, warming-induced water stress compromised the NPP in arid sparse grassland and deserts. Cropland had greater NPP and NPP increase than natural vegetation due to the irrigation, fertilizers, and other artificial inputs it received. The decrease in both temperature and precipitation generally reduced NPP, but the NPP in the well-protection or less-disturbance areas still increased slightly.


The connection between dystocia and dysmenorrhoea.
Eight hundred and eighty women having their first baby were asked whether they had had dysmenorrhoea; 61 per cent had and 39 per cent had not. Eleven per cent of the former had to have forceps, ventouse or caesarean section, compared with twice as many (20.8 per cent) of the latter (p = 0.001). Those with severe dysmenorrhoea had a greater need for operative interference (not statistically significant). I suggest, after 21 years of observation, that the level of prostaglandin activity may be different in these women, and that a history of no dysmenorrhoea or severe dysmenorrhoea might be considered as a risk factor for a complicated delivery.


A novel PKB/Akt inhibitor, MK-2206, effectively inhibits insulin-stimulated glucose metabolism and protein synthesis in isolated rat skeletal muscle.
PKB (protein kinase B), also known as Akt, is a key component of insulin signalling. Defects in PKB activation lead to insulin resistance and metabolic disorders, whereas PKB overactivation has been linked to tumour growth. Small-molecule PKB inhibitors have thus been developed for cancer treatment, but also represent useful tools to probe the roles of PKB in insulin action. In the present study, we examined the acute effects of two allosteric PKB inhibitors, MK-2206 and Akti 1/2 (Akti) on PKB signalling in incubated rat soleus muscles. We also assessed the effects of the compounds on insulin-stimulated glucose uptake, glycogen and protein synthesis. MK-2206 dose-dependently inhibited insulin-stimulated PKB phosphorylation, PKBβ activity and phosphorylation of PKB downstream targets (including glycogen synthase kinase-3α/β, proline-rich Akt substrate of 40 kDa and Akt substrate of 160 kDa). Insulin-stimulated glucose uptake, glycogen synthesis and glycogen synthase activity were also decreased by MK-2206 in a dose-dependent manner. Incubation with high doses of MK-2206 (10 μM) inhibited insulin-induced p70 ribosomal protein S6 kinase and 4E-BP1 (eukaryotic initiation factor 4E-binding protein-1) phosphorylation associated with increased eEF2 (eukaryotic elongation factor 2) phosphorylation. In contrast, Akti only modestly inhibited insulin-induced PKB and mTOR (mammalian target of rapamycin) signalling, with little or no effect on glucose uptake and protein synthesis. MK-2206, rather than Akti, would thus be the tool of choice for studying the role of PKB in insulin action in skeletal muscle. The results point to a key role for PKB in mediating insulin-stimulated glucose uptake, glycogen synthesis and protein synthesis in skeletal muscle.


Tibial lengthening for unilateral Crowe type-IV developmental dysplasia of the hip.
Developmental dysplasia of the hip (DDH) is associated with chronic pain and limping which especially has a negative impact on the patients' daily activities, body image, and self-esteem. Although total hip arthroplasty remains the first choice for treatment of DDH in adults, minimally invasive alternative approaches are being increasingly favored both by the surgeon and the patients with severe DDH. This study aimed to evaluate the outcome of these patients treated with a mono-lateral external fixator-based tibial lengthening procedure. During the period of month between June 1999 and January 2006, 13 (mean ages 20.8 years) adult patients with unilateral Crowe type-IV DDH were treated by tibial lengthening using a mono-lateral external fixator over an intramedullary nail. Bone healing, infection, gait correction and improvement in body image were assessed during postoperative followup. Patients' overall health status at the end of followup was assessed using the short form-36 (SF-36) health survey. Patients were followed up for an average of 7.3 years. Successful bone healing was observed in all 13 patients and no further surgeries were indicated. A mean external fixation index of 12.4 days/cm was achieved. Bone formation fell in good to excellent categories with a mean consolidation index of 50.1 days/cm. Pin-tract infections were observed in two patients. The degree of limping was reduced from severe or moderate preoperatively to mild postoperatively. Neither equinus deformity nor painful degenerative osteoarthritis and hip dysfunction were observed in any of the patients studied. The SF-36 questionnaire survey showed that all patients were satisfied with their outcomes. Tibial lengthening may effectively correct gait and satisfactorily improve body image in young patients with unilateral Crowe type-IV DDH. Mono-lateral external fixator allows for accelerated postoperative rehabilitation and optimal preservation of ankle movements. Lengthening along with intramedullary nails may significantly reduce the external fixation time and the risk of fixator-related complications.


Demographics, diagnostics, treatment, and outcomes of patients presenting with acute groin hernia: 15-year multicentre retrospective cohort study.
Groin hernias commonly present acutely in high-risk populations and can be challenging to manage. This retrospective, observational study aimed to report on patient demographics and outcomes, following acute admissions with a groin hernia, in relation to contemporary investigative and management practices. Adult (≥18 years old) patients who presented acutely with a groin hernia to nine National Health Service trusts in the north of England between 2002 and 2016 were included. Data were collected regarding patient demographics, radiological investigations, and operative intervention. The primary outcome of interest was 30-day inpatient mortality rate. Overall, 6165 patients with acute groin hernia were included (4698 inguinal and 1467 femoral hernias). There was a male preponderance (72.5 per cent) with median age of 73 years (interquartile range (i.q.r.) 58-82). The burden of patient co-morbidity increased over the study period (P < 0.001). Operative repair was performed in 2258 (55.1 per cent) of patients with an inguinal and 1321 (90.1 per cent) of patients with a femoral hernia. Bowel resection was more commonly required for femoral hernias (14.7 per cent) than inguinal hernias (3.5 per cent, P < 0.001) and in obstructed (14.6 versus 0.2 per cent, P < 0.001) or strangulated (58.4 versus 4.5 per cent, P < 0.001) hernias. The 30-day mortality rate was 3.1 per cent for the overall cohort and 3.9 per cent for those who underwent surgery. Bowel resection was associated with increased duration of hospital stay (P < 0.001) and 30-day inpatient mortality rate (P < 0.001). Following adjustment for confounding variables, advanced age, co-morbidity, obstruction, and strangulation were all associated with an increased 30-day mortality rate (all P < 0.001). Emergency hernia repair has high mortality rates. Advanced age and co-morbidity increase both duration of hospital stay and 30-day mortality rate.


Peripheral neuropathy induced by microtubule-stabilizing agents.
Microtubule-stabilizing agents (MTSAs), including the taxanes and epothilones, are effective chemotherapeutic agents for the treatment of many cancers. Neuropathy is a major adverse effect of MTSA-based chemotherapy, with severe peripheral neuropathy (grade 3 or 4) occurring in as many as 30% of patients treated with a MTSA. MTSA-induced neuropathy usually resolves gradually after cessation of the treatment. The most reliable method to accurately assess MTSA-induced neuropathy is by clinical evaluation, although additional techniques are being developed and evaluated. Among MTSA-induced neuropathy, the most extensively studied is that induced by taxanes; such a neuropathy usually presents as sensory neuropathy and is more common with paclitaxel than docetaxel. The incidence of MTSA-induced neuropathy seems to depend on the MTSA dose per treatment cycle, the schedule of treatment, and the duration of the infusion. Although there have been several small clinical trials with neuroprotective agents, early recognition and supportive care are the best approaches for prevention and management of MTSA-induced neuropathy. In the future, research should focus on elucidating the mechanism of MTSA-induced neuropathy, developing reliable in vivo and in vitro preclinical models to study MTSA-induced neuropathy, developing a more reliable grading system for MTSA-induced neuropathy, developing more reliable methods for evaluating MTSA-induced neuropathy, and evaluating the efficacy of potential neuroprotective agents in clinical trials.


Genetic Polymorphism Effect on Warfarin-Rifampin Interaction: A Case Report and Review of Literature.
Warfarin-rifampin interaction has been reported since the 1970s. Due to rifampin's strong induction of CYP2C9, most cases could not attain the target international normalized ratio (INR) despite warfarin dose escalation. Genetic polymorphisms determine up to 50% of warfarin dose variability. A 38-year-old woman was started on warfarin and rifampin for cerebral venous sinus thrombosis and pulmonary tuberculosis. Over six weeks, the daily warfarin dose was increased from 3 to 10 mg to attain three consecutive in-clinic therapeutic INRs. She completed three complications-free months of warfarin treatment with time in therapeutic range (TTR) of 46%. We performed retrospective genetic testing to determine the patient's CYP2C9, CYP4F2, and VKORC1 genotypes and whether they had affected the interaction outcome. The analysis revealed that the subject carries CYP2C9*3*3 and VKORC1-1639 (GA) mutations, classifying her as a slow metabolizer and, hence, highly warfarin-sensitive. This was reflected on how the case responded to a relatively lower dose than previously reported cases that did not achieve the target on warfarin daily doses up to 35 mg. This is the first report addressing the genotype effect on this interaction. Patients with genetic variants requiring low warfarin doses are more likely to respond at a feasible dose while on rifampin. Future studies to evaluate warfarin-rifampin-gene interaction are warranted.


Bone turnover markers: understanding their value in clinical trials and clinical practice.
While bone mineral density (BMD) by dual-energy X-ray absorptiometry is the primary method of determining fracture risk, assessing bone turnover may add valuable information for the management of patients with low bone mass. Bone turnover markers (BTMs) are used in clinical trials where they can provide essential information on the biological efficacy of osteoporosis treatments. In such population-based studies, BTMs can predict fracture risk independent of BMD. When combined with BMD, they improve the fracture risk estimate above and beyond BMD alone in postmenopausal osteoporotic women. Since changes in bone turnover after the initiation of therapy with bone resorption inhibitors occur much more rapidly than changes in BMD, treatment efficacy could, in theory, be determined within weeks of using BTMs. However, such predictive value is limited by the large biological variability of these biochemical markers, even though newer automated methods have reduced their analytical variability. Consequently, widespread adoption as a means of predicting treatment efficacy in fracture prevention for individual patients cannot yet be recommended. BTMs may be useful for monitoring adherence to antiresorptive therapy and may aid in identifying patients for whom antiresorptive therapy is most appropriate. Thus, although BTMs are currently confined to clinical research applications, further improvement in assay precision may extend their diagnostic value in clinical settings.


Pathways of introduction of the invasive aquatic plant Cabomba caroliniana.
The pathway and frequency of species' introductions can affect the extent, impact, and management of biological invasions. Here, we examine the pathway of introduction of the aquatic plant Cabomba caroliniana (fanwort) into Canada and the northern United States using plastid DNA sequence (intergenic spacers atpF-atpH, trnH-psbA, and trnL-trnF) and DNA content analyses. We test the hypothesis that the spread of fanwort is a result of commercial trade by comparing a Canadian population (Kasshabog Lake, ON) to native populations from southern U.S., introduced populations in northern U.S., and plants from commercial retailers. Thirteen plastid haplotypes were identified throughout North America, including one dominant haplotype, which was present in all C. caroliniana populations. Several rare haplotypes were used to infer shared colonization history. In particular, the Canadian population shared two rare alleles with a population from Massachusetts, suggesting range expansion of C. caroliniana from the northern U.S. However, the possibility of a commercial introduction cannot be excluded, as common alleles were shared between the Canadian population and both commercial and southern U.S. sources. Variation in C. caroliniana genome size was bimodal and populations were classified into "high" and "low" categories. The Canadian population had DNA contents similar to several northern U.S. populations (low DNA content). This may provide additional support for range expansion from these introduced populations rather than from commercial sources or populations in the southern U.S., which had high DNA content.


Patterns of motorcycle helmet use - A naturalistic observation study in Myanmar.
Developing countries are subject to increased motorization, particularly in the number of motorcycles. As helmet use is critical to the safety of motorcycle riders, the goal of this study was to identify observable patterns of helmet use, which allow a more accurate assessment of helmet use in developing countries. In a video based observation study, 124,784 motorcycle riders were observed at seven observation sites throughout Myanmar. Recorded videos were coded for helmet use, number of riders on the motorcycle, rider position, gender, and time of day. Generally, motorcycle helmet use in Myanmar was found to be low with only 51.5% percent of riders wearing a helmet. Helmet use was highest for drivers (68.1%) and decreased for every additional passenger. It was lowest for children standing on the floorboard of the motorcycle (11.3%). During the day, helmet use followed a unimodal distribution, with the highest use observed during the late morning and lowest use observed in the early morning and late afternoon. Helmet use varied significantly between observation sites, ranging from 74.8% in Mandalay to 26.9% in Pakokku. In Mandalay, female riders had a higher helmet use than male riders, and helmet use decreased drastically on a national holiday in the city. Helmet use of motorcycle riders in Myanmar follows distinct patterns. Knowledge of these patterns can be used to design more precise helmet use evaluations and guide traffic law policy and police enforcement measures. Video based observation proved to be an efficient tool to collect helmet use data.


Altered placental lactogen and leptin expression in placentomes from bovine nuclear transfer pregnancies.
Appropriate growth, development, and function of the placenta is central to the success of nutrient partitioning between the mother, placenta, and fetus. Hormones such as placental lactogen (PL) and leptin are produced in the bovine placenta and play an important role in nutrient partitioning and regulation of placental and fetal growth. Nuclear transfer pregnancies are associated with a number of fetal and placental abnormalities, including increased placental growth and macrosomia, and hence represent a unique situation to gain insight into fetoplacental growth regulation. We have examined the expression of bovine PL (bPL) and leptin in placentomes of artificially inseminated (AI), in vitro produced (IVP), and nuclear transfer (NT) pregnancies at Days 50, 100, and 150 of gestation in the cow. Immunolocalization studies showed that spatial and temporal patterns of expression of bPL and leptin were markedly altered in the placentomes of NT pregnancies compared with AI or IVP controls. Concentrations of bPL in allantoic fluid, as determined by radioimmunoassay (RIA), were significantly higher (P < or = 0.001) in NT pregnancies (17.9 +/- 3.2 ng/ml; mean +/- SD) compared with AI (2.03 +/- 1.5 ng/ml), but not IVP (23.4 +/- 12.8 ng/ ml) pregnancies on Day 150 of gestation. In contrast, amniotic fluid levels of bPL were significantly decreased in NT pregnancies at Day 150 gestation. Leptin mRNA expression, as determined by real-time reverse transcription-PCR, was increased 2.4- to 3.0-fold in NT placentomes compared with AI controls at all gestational ages examined. We speculate that the observed dysregulation of expression of bPL and leptin in NT placentomes could contribute to aberrations in cell migration and invasion and subsequently to alterations in placental metabolism and transfer of nutrients to the fetus, thus leading to increased placental and fetal macrosomia in NT pregnancies.


Hemodynamic and metabolic changes caused by regional ischemia in porcine heart.
Myocardial ischemia was induced in the pig by reducing the left anterior descending coronary artery (LAD) flow to 26% of its control value. After one hour, the LAD was reperfused for 50 minutes. During ischemia and reperfusion, cardiac output correlated strongly with arterial levels of lactate and nucleosides (all r less than - 0.85). Myocardial inosine production seems to be a good marker of ischemia because it correlated very well with myocardial lactate production (r = 0.99).


Pharmacological interventions for paranoid personality disorder.
This is the protocol for a review and there is no abstract. The objectives are as follows: To evaluate the effects of pharmacological interventions for people with paranoid personality disorder (PPD).


Meningomyelocele and syringohydromyelia. Some radiological aspects.
Twenty meningomyelocele patients with nonfunctional ventricular shunts or no shunts were studied radiologically. This group was compared to a second group of 4 patients with functional shunts. The various techniques are compared and discussed. Pantopaque ventriculography is a potentially dangerous procedure. Radioisotope ventriculography appears to be a safe and useful method of diagnosing syringohydromyelia. Ventricular decomposition by means of a shunt appears to be an effective treatment.


Patient education about cough: effect on the consulting behaviour of general practice patients.
The aim of this general practice study was to examine how the consulting behaviour of patients with a cough was affected when the tasks and responsibilities of patients, practice nurses and general practitioners were reorganized. In four 'average' single-handed general practices the effects on the consulting behaviour of patients of a rational practice policy on cough and the provision of systematic patient education on cough were compared with patient behaviour in four matched control practices. Changes of behaviour were measured in 548 patients who consulted for cough at least twice, in two successive autumn-winter periods. Significantly more patients in the experimental practice changed their behaviour to follow the practice guidelines than did patients in the control practices (56% versus 30%, P less than 0.001). The proportion of patients who continued to consult in the approved manner was greater among patients receiving intervention (66% versus 29%, P less than 0.001). This was equally true for patients who had suffered less than four episodes of cough or more than four episodes. The more often the patients received the education, the more effective it was. All patients who consulted the general practitioner for cough during the first autumn-winter period filled in a cough diary during the second period. From this it appeared that the intervention did not result in patients delaying consultation when they had a cough lasting longer than three weeks or one with 'serious' symptoms. It would appear that a rational practice policy and the provision of patient education can stimulate patients to modify their consulting behaviour. This could result in a reduction in the costs of health care.


Dynamic electro-optic response of graphene/graphitic flakes in nematic liquid crystals.
Electric field induced dynamic reorientation phenomenon of graphene/graphitic flakes in homogeneously aligned nematic liquid crystal (NLC) medium has been demonstrated by optical microscopy. The flakes reorient from parallel to perpendicular configuration with respect to boundary plates of confining cells for an applied field strength of as low as tens of millivolt per micrometer. After field removal the reoriented flakes recover to their initial state with the help of relaxation of NLC. Considering flake reorientation phenomenon both in positive and negative dielectric anisotropy NLCs, the reorientation process depends on interfacial Maxwell-Wagner polarization and NLC director reorientation. We propose a phenomenological model based on electric field induced potential energy of graphitic flakes and coupling contribution of positive NLC to generate the rotational kinetic energy for flake reorientation. The model successfully explains the dependence of flake reorientation time over flake shape anisotropy, electric-field strength, and flake area. Using present operating scheme it is possible to generate dark field-off state and bright field-on state, having application potential for electro-optic light modulation devices.


Physical assessment. The heart.
This article discusses normal and abnormal anatomy and physiology of the heart and the techniques used in physical assessment. Specific adverse drug effects are discussed and a case study is provided to illustrate key concepts.


Management of primary and metastatic spinal tumors.
ìTumors of the spine, though rare, may significantly impact quality of life. The management depends not only on clinical presentation, but also on histology, stage and grade of the tumor. Primary benign tumors usually are a focal problem, however, may be focally aggressive. Primary malignant tumors are always considered aggressive and are managed in a multidisciplinary fashion. Surgical treatment often requires aggressive en bloc resection to maximize potential for cure. Metastatic tumors of the spine may cause a significant impact on the quality of life. In select cases with spine instability, pain and neurologic deficit surgical management is indicated.


16S-23S rRNA Internal Transcribed Spacer Region (ITS) Sequencing: A Potential Molecular Diagnostic Tool for Differentiating Lactococcus garvieae and Lactococcus petauri.
Lactococcus garvieae is the etiological agent of lactococcosis, a clinically and economically significant infectious disease affecting farmed rainbow trout. L. garvieae had been considered the only cause of lactococcosis for a long time; however, L. petauri, another species of the genus Lactococcus, has lately been linked to the same disease. The genomes and biochemical profiles of L. petauri and L. garvieae have a high degree of similarity. Traditional diagnostic tests currently available cannot distinguish between these two species. The aim of this study was to use the transcribed spacer (ITS) region between 16S rRNA and 23S rRNA as a potential useful molecular target to differentiate L. garvieae from L. petauri, saving time and money compared to genomics methods currently used as diagnostic tools for accurate discrimination between these two species. The ITS region of 82 strains was amplified and sequenced. The amplified fragments varied in size from 500 to 550 bp. Based on the sequence, seven SNPs were identified that separate L. garvieae from L. petauri. The 16S-23S rRNA ITS region has enough resolution to distinguish between closely related L. garvieae and L. petauri and it can be used as a diagnostic marker to quickly identify the pathogens in a lactococcosis outbreak.