Impact of diabetes on heart failure incidence in adults with ischemic heart disease.
Ischemic heart disease (IHD) is the most potent risk factor for heart failure (HF). Our study aims to evaluate the incremental impact of diabetes on the incidence of HF in individuals with IHD. Data from the NHANES Epidemiologic Follow-Up Study (Baseline: 1971 to 1974) were linked to the facility and mortality files up to 1992. Our analyses were restricted to patients with IHD without prevalent HF at baseline. The cumulative incidence of HF in patients with diabetes and IHD versus those with IHD alone was assessed using failure curves. Cox proportional hazards models were used to control for important covariates. All analyses incorporated the complex sample design by including the weights and clustering variables. Out of the 14,407 participants, 497 had IHD without prevalent HF and had information about diabetes status. Among these participants, the cumulative incidence of HF was 38.1% for those with diabetes (n=63) and 26.5% in those without diabetes (n=434) (log-rank p-value<0.005). The multivariate hazard ratio (adjusted for age, BMI, alcohol consumption, hypertension, high cholesterol, and smoking) for incident HF for people who had myocardial infarction (MI) and diabetes compared to people who had MI alone was 2.98 (95% CI 1.51, 5.88). Among participants with MI, those with diabetes had a substantially higher incidence of HF than those without diabetes. Based on these findings, practitioners should focus greater attention on patients with diabetes and previous MI in order to potentially prevent incident HF.


Sheltered versus nonsheltered homeless women differences in health, behavior, victimization, and utilization of care.
To contrast sociodemographic characteristics, physical and mental health status, substance use, sexual behaviors, victimization, and utilization of health services between homeless women residing in sheltered and non-sheltered environments. Cross-sectional survey. A structured scale was used to measure mental health status. Physical health status, substance use, sexual behavior, history of adult victimization, and health services utilization were measured by content-specific items. Shelters (N = 47) and outdoor locations in Los Angeles. One thousand fifty-one homeless women. Homeless women living on the streets were more likely than sheltered women to be white and longer-term homeless. Controlling for sociodemographic characteristics, multiple logistic regression analyses revealed that unsheltered women had over 3 times greater odds of fair or poor physical health, and over 12 times greater odds of poor mental health than sheltered homeless women. They were also more likely than sheltered women to report using alcohol or noninjection drugs, to have multiple sexual partners, and to have a history of physical assault. About half of the overall sample reported utilization of a variety of health services; however, unsheltered homeless women were less likely to utilize all of the health services that were assessed, including drug treatment. There is a critical need for aggressive outreach programs that provide mental health services and substance abuse treatment for homeless women on the streets. Comprehensive services that also include medical care, family planning, violence prevention, and behavioral risk reduction may be particularly valuable for homeless women, especially those living in unsheltered environments.


Photothermal-assisted surface-mediated gene delivery for enhancing transfection efficiency.
The development of gene therapy puts forward the requirements for efficient delivery of genetic information into diverse cells. However, in some cases of transfection, especially those for transfecting some primary cells and for delivering large size plasmid DNA (pDNA), the existing conventional transfection methods show poor efficiency. How to further improve transfection efficiency in these hard-to-achieve issues remains a crucial challenge. Here, we report a photothermal-assisted surface-mediated gene delivery based on a polydopamine-polyethylenimine (PDA-PEI) surface. The PDA-PEI surface was prepared through PEI-accelerated dopamine polymerization, which showed efficiency in the immobilization of PEI/pDNA polyplexes and remarkable photothermal properties. Upon IR irradiation, we observed improved transfection efficiencies of two important hard-to-achieve transfection issues, namely the transfection of primary endothelial cells, which are kinds of typical hard-to-transfect cells, and the transfection of cells with large-size pDNA. We demonstrate that the increases of transfection efficiency were due to the hyperthermia-induced pDNA release, the local cell membrane disturbance, and the polyplex internalization. This work highlights the importance of local immobilization and release of pDNA to gene deliveries, showing great potential applications in medical devices in the field of gene therapy.


Molecular weight of detergent-solubilized prostaglandin-F2 alpha receptor from bovine corpora lutea.
A prostaglandin F2 alpha receptor localized in plasma membranes of bovine corpus luteum cells was solubilized by treatment with Triton X-100. Sepharose chromatographies of ([3H]prostaglandin F2 alpha)-receptor complex gave a Stokes' radius of 630 nm. In the absence of detergent, aggregated forms of the receptor appeared. Sedimentation experiments of solubilized receptor in sucrose/H2O and sucrose/2H2O density gradients gave the following values: sedimentation coefficient (S20, w) 4.6 S; partial specific volume (VB) 0.78 cm3/g and frictional ratio (f/fo) 1.6. Based on the sedimentation coefficient and the Stokes' radius and assuming that the receptor is a non-glycosylated protein the molar mass of the receptor-(Triton X-100) complex was 144000 g/mol. The VB value indicated that ca. 26% of the weight represented bound detergent and that the molecular weight of the prostaglandin F2 alpha receptor is approximately 107000.


Protective role of curcumin on lindane induced reproductive toxicity in male Wistar rats.
The aim of this study was to explore the adverse effects of lindane pesticide on testes and epididymus weight, sperm head counts, sperm motility, abnormal changes in sperm morphology, biochemical changes in endogenous antioxidants and oxidative enzymes in male wistar rats as well as to assess ameliorating role of 'curcumin'. Rats were exposed daily to lindane (30 mg/kg bw) for 14 and 28 days and administered with curcumin (100 mg/kg bw) in pretreatment, post treatment and combination groups. We observed decrease (p < 0.001) in testes and cauda epididymus weight, decrease (p < 0.001) in testicular sperm head count, increase (p < 0.001) in abnormal tail morphology (headless tail, multiple tail, broken tail, coiled tail and bent tail), abnormal head morphology (excessive hook, amorphous head, pin head, short head, blunt hook and detached hook) and decrease (p < 0.001) in sperm motility after lindane exposure, as compared to control. There was (p < 0.01) decline in superoxide dismutase, catalase and glutathione-s-transferase activity on lindane exposure, however, no change (p > 0.05) was observed in glutathione level. Lipid peroxidation was (p < 0.01) enhanced on lindane exposure as compared to control. Curcumin administration was able to ameliorate lindane induced reproductive toxicity in pretreatment, post treatment and combination groups. The study indicated that even after 14 days of metabolism, toxic effects of lindane were evident.


Effect of changes in thyroid status on tissue respiration in fetal and newborn sheep.
The effect of both exogenous and endogenous changes in thyroid status on in vitro tissue respiration and thermogenic enzymes (sodium-potassium-adenosine triphosphatase and alpha-glycerophosphate dehydrogenase) was studied in fetal and newborn sheep. Oxygen consumption of liver and brain increased from 25 +/- 4.1 and 58.5 +/- 2.8 microliters O2 X 100 mg-1 X h-1, respectively, in tissues from unthyroidectomized fetal animals at 136-140 days gestation to 60 +/- 4.2 and 72 +/- 1.5 microliters O2 X 100 mg-1 X h-1 in tissues from unthyroidectomized newborn lambs between birth and 7 days of age. The physiological changes in thyroid function that normally occur at birth resulted in a mean (+/- SE) plasma triiodothyronine (T3) concentration of 563 +/- 39 ng/dl in the newborn lambs compared with 39 +/- 8 ng/dl in the fetal animals. Kidney respiration and thermogenic enzyme activities in the several tissues studied did not change. Liver, kidney, and brain respiration and thermogenic enzymes from T3-treated thyroidectomized fetal and newborn lambs were not increased (compared with untreated thyroidectomized animals) despite a marked increase in plasma T3 concentrations. Conclusions are that 1) liver, kidney cortex, and frontal brain cortex in the fetal and newborn lamb are relatively insensitive to the calorigenic effect of thyroid hormones, and 2) a perinatal increase in hepatic and cerebral respiration occurs in newborn animals (compared with fetal animals) but is probably not due solely to perinatal increases in thyroid hormones.


Comparison of the efficacy of ramoplanin and vancomycin in both in vitro and in vivo models of clindamycin-induced Clostridium difficile infection.
Treatment of Clostridium difficile infection (CDI) is limited primarily to either metronidazole or vancomycin. We compared vancomycin and a novel glycolipodepsipeptide, ramoplanin, in both hamster and in vitro gut models of clindamycin-induced CDI. We used an in vitro triple-stage chemostat model that simulates the human gut, and an in vivo hamster model, both primed with clindamycin. Clindamycin exposure elicited symptomatic disease in the hamster model, and promoted C. difficile germination and toxin production in the gut model. C. difficile germination and toxin production were not associated with depletion of gut microflora in the gut model, but were temporarily associated with subinhibitory concentrations of clindamycin. Both ramoplanin and vancomycin were associated with rapid symptom resolution in the hamster model, and rapid toxin titre decrease in the in vitro gut model. In both models of CDI, vancomycin was associated with greater persistence of C. difficile spores. C. difficile spores were recovered significantly more often from the caecal contents of vancomycin-treated (n = 19/23) compared with ramoplanin-treated (n = 6/23) hamsters (P < 0.05). Results from the in vitro gut and hamster models were concordant. Ramoplanin and vancomycin were similarly effective at reducing cytotoxin production in the gut CDI model and in resolving symptoms in the hamster model. Ramoplanin may be more effective than vancomycin at killing spores and preventing spore recrudescence. These findings suggest a potential therapeutic role for ramoplanin in CDI that requires further clinical investigation.


Evidence of aquaporin involvement in human central pontine myelinolysis.
Central pontine myelinolysis (CPM) is a demyelinating disorder of the central basis pontis that is often associated with osmotic stress. The aquaporin water channels (AQPs) have been pathogenically implicated because serum osmolarity changes redistribute water and osmolytes among various central nervous system compartments. We characterized the immunoreactivity of aquaporin-1 and aquaporin-4 (AQP1 and AQP4) and associated neuropathology in microscopic transverse sections from archival autopsied pontine tissue from 6 patients with pathologically confirmed CPM. Loss of both AQP1 and AQP4 was evident within demyelinating lesions in four of the six cases, despite the presence of glial fibrillary acidic protein (GFAP)-positive astrocytes. Lesional astrocytes were small, and exhibited fewer and shorter processes than perilesional astrocytes. In two of the six cases, astrocytes within demyelinating lesions exhibited increased AQP1 and AQP4 immunoreactivities, and gemistocytes and mitotic astrocytes were numerous. Blinded review of medical records revealed that all four cases lacking lesional AQP1 and AQP4 immunoreactivities were male, whereas the two cases with enhanced lesional AQP1 and AQP4 immunoreactivities were female. This report is the first to establish astrocytic AQP loss in a subset of human CPM cases and suggests AQP1 and AQP4 may be involved in the pathogenesis of CPM. Further studies are required to determine whether the loss of AQP1 and AQP4 is restricted to male CPM patients, or rather may be a feature associated with specific underlying precipitants of CPM that may be more common among men. Non-rodent experimental models are needed to better clarify the complex and dynamic mechanisms involved in the regulation of AQPs in CPM, in order to determine whether it occurs secondary to the destructive disease process, or represents a compensatory mechanism protecting the astrocyte against apoptosis.


Sphingosine 1-phosphate, a potential target in neovascular retinal disease.
Neovascular ocular diseases (such as age-related macular degeneration, diabetic retinopathy and retinal vein occlusion) are characterised by common pathological processes that contribute to disease progression. These include angiogenesis, oedema, inflammation, cell death and fibrosis. Currently available therapies target the effects of vascular endothelial growth factor (VEGF), the main mediator of pathological angiogenesis. Unfortunately, VEGF blockers are expensive biological therapeutics that necessitate frequent intravitreal administration and are associated with multiple adverse effects. Thus, alternative treatment options associated with fewer side effects are required for disease management. This review introduces sphingosine 1-phosphate (S1P) as a potential pharmacological target for the treatment of neovascular ocular pathologies. S1P is a sphingolipid mediator that controls cellular growth, differentiation, survival and death. S1P actions are mediated by five G protein-coupled receptors (S1P1-5 receptors) which are abundantly expressed in all retinal and subretinal structures. The action of S1P on S1P1 receptors can reduce angiogenesis, increase endothelium integrity, reduce photoreceptor apoptosis and protect the retina against neurodegeneration. Conversely, S1P2 receptor signalling can increase neovascularisation, disrupt endothelial junctions, stimulate VEGF release, and induce retinal cell apoptosis and degeneration of neural retina. The aim of this review is to thoroughly discuss the role of S1P and its different receptor subtypes in angiogenesis, inflammation, apoptosis and fibrosis in order to determine which of these S1P-mediated processes may be targeted therapeutically.


Substance P and Substance P receptor antagonists in the pathogenesis and treatment of affective disorders.
Substance P (SP) is a neuropeptide which is widely distributed in the periphery and the central nervous system (CNS), where it is co-localised with other neurotransmitters such as serotonin or dopamine and where it acts as a neuromodulator. SP has been proposed to play a role in the aetiopathology of asthma, inflammatory bowel disease, emesis, psoriasis, as well as neuropsychiatric disorders including pain syndromes (e.g. migraine and fibromyalgia) and affective disorders, anxiety disorders, schizophrenia and Alzheimer's disease. This review focuses on the role of SP in the pathogenesis of affective disorders. It summarises the current knowledge on measurements of SP in the CSF and serum in patients with depressive disorders or fibromyalgia, effects of SP-application in humans, SP-receptor expression in postmortem brains and the modulation of SP levels in the course of antidepressant treatment. It also discusses the promise of substance P-receptor antagonists (SPA) for the treatment of affective disorders and their proposed mechanism of action. In summary, much more research is needed to elucidate the role of SP in the pathogenesis of depression. SPA are promising as future drugs for the treatment of affective disorders, but current clinical trials have yet to be completed to draw a firm conclusion. Key words: substance P, neurokinin1-receptor, affective disorders, depression, review.


Squeezing, then stacking: from breathing pores to three-dimensional ionic self-assembly under electrochemical control.
We demonstrate the spontaneous and reversible transition between the two- and three-dimensional self-assembly of a supramolecular system at the solid-liquid interface under electrochemical conditions, using in situ scanning tunneling microscopy. By tuning the interfacial potential, we can selectively organize our target molecules in an open porous pattern, fill these pores to form an auto-host-guest structure, or stack the building blocks in a stratified bilayer. Using a simple electrostatic model, we rationalize which charge density is required to enable bilayer formation, and conversely, which molecular size/charge ratio is necessary in the design of new building blocks. Our results may lead to a new class of electrochemically controlled dynamic host-guest systems, artificial receptors, and smart materials.


Prognostic factors in children with severe diffuse brain injuries: a study of 74 patients.
Severe diffuse brain injury in children has a devastating influence on their physical and psychological development. This retrospective study was undertaken to analyse the factors that influence outcome in children with such injuries. The short-term outcomes (Glasgow outcome score) of 74 children (age < or =15 years) with severe diffuse brain injury and no focal operable mass lesions on CT scan, admitted between 1992 and 1998 at the National Institute of Mental Health and Neurosciences were analysed. The prognostic factors analysed were age, gender, time after injury to admission, nature of injury, highest post-resuscitation Glasgow coma score (GCS), pupillary reaction, horizontal oculocephalic reflex and CT scan findings. The following factors were found to be independent predictors of poor outcome: GCS 3-5 (p < 0.003), absent verbal response (p < 0.001), motor response score of 1-3 (p < 0.001), absent pupillary reaction (p < 0.001), absent oculocephalic reflex (p < 0.001) and presence of traumatic subarachnoid haemorrhage on CT scan (p < 0.002). These independent variables were then subjected to a stepwise logistic regression analysis, and the most important variables for predicting outcome were oculocephalic reflex and GCS, which together correctly predicted unfavourable outcome with a sensitivity of 79% and a specificity of 65%. An early CT scan or a single CT scan did not have any prognostic significance.


Aggregation of human S100A8 and S100A9 amyloidogenic proteins perturbs proteostasis in a yeast model.
Amyloid aggregates of the calcium-binding EF-hand proteins, S100A8 and S100A9, have been found in the corpora amylacea of patients with prostate cancer and may play a role in carcinogenesis. Here we present a novel model system using the yeast Saccharomyces cerevisiae to study human S100A8 and S100A9 aggregation and toxicity. We found that S100A8, S100A9 and S100A8/9 cotransfomants form SDS-resistant non-toxic aggregates in yeast cells. Using fluorescently tagged proteins, we showed that S100A8 and S100A9 accumulate in foci. After prolonged induction, S100A8 foci localized to the cell vacuole, whereas the S100A9 foci remained in the cytoplasm when present alone, but entered the vacuole in cotransformants. Biochemical analysis of the proteins indicated that S100A8 and S100A9 alone or coexpressed together form amyloid-like aggregates in yeast. Expression of S100A8 and S100A9 in wild type yeast did not affect cell viability, but these proteins were toxic when expressed on a background of unrelated metastable temperature-sensitive mutant proteins, Cdc53-1p, Cdc34-2p, Srp1-31p and Sec27-1p. This finding suggests that the expression and aggregation of S100A8 and S100A9 may limit the capacity of the cellular proteostasis machinery. To test this hypothesis, we screened a set of chaperone deletion mutants and found that reducing the levels of the heat-shock proteins Hsp104p and Hsp70p was sufficient to induce S100A8 and S100A9 toxicity. This result indicates that the chaperone activity of the Hsp104/Hsp70 bi-chaperone system in wild type cells is sufficient to reduce S100A8 and S100A9 amyloid toxicity and preserve cellular proteostasis. Expression of human S100A8 and S100A9 in yeast thus provides a novel model system for the study of the interaction of amyloid deposits with the proteostasis machinery.


Risk Factors of Stomal Recurrence After Laryngectomy: A Systematic Review and Meta-analysis.
We wished to investigate the risk factors for stoma recurrence following laryngectomy. PubMed, Cochrane Library, and Embase were searched to identify primary research studies published between January 1, 1967, and October 31, 2016. We only included observational epidemiological studies and used risk ratios (RRs) and 95% confidence intervals (CI) to summarize the primary risk factors associated with recurrence following laryngectomy. A total of 44 articles, including 11 928 patients, were included in the current meta-analysis. The pooled estimates of the stomal recurrence rate and the local recurrence rate following laryngectomy were 6.60% (95% CI, 5.40-7.90) and 19.40% (95% CI, 14.00%-24.80%), respectively. For stomal recurrence, we confirmed a series of earlier identified factors, including tumor site, prior tracheotomy, tracheotomy timing, T-classification, lymph node metastases, postoperative pharyngoperistomal fistula, and a positive surgical margin. For local recurrence, postoperative radiotherapy, overexpression of p53, and overexpression of p21 were significant, while preoperative tracheostomy and postoperative pharyngoperistomal fistula had an unexpectedly low impact on risk. This study has confirmed a series of earlier identified factors for stomal recurrence and local recurrence following laryngectomy. Our results will provide important insights for clinical practice.


Validation of the Block Walk Method for Assessing Physical Activity occurring on Sidewalks/Streets.
The block walk method (BWM) is one of the more common approaches for assessing physical activity (PA) performed on sidewalks/streets; however, it is non-technical, labor-intensive, and lacks validation. This study aimed to validate the BWM and examine the potential for using a wearable video device (WVD) to assess PA occurring on sidewalks/streets. Trained observers (one wearing and one not wearing the WVD) walked together and performed the BWM according to a previously developed protocol along routes in low, medium, and high walkable areas. Two experts then reviewed the videos. A total of 1150 (traditional) and 1087 (video review) individuals were observed during 900 min of observation. When larger numbers of individuals were observed, the traditional method overestimated the overall number of people as well as those walking and sitting/standing, while underestimating the number of runners. Valid estimates of PA occurring on sidewalks/streets can be obtained by the traditional BWM in low and medium walkability areas and/or with non-common activities (cycling); however, its validity is questionable when sidewalks/streets use volume is high. The use of WVDs in PA assessment has the potential to establish new levels of accuracy, reduce resource requirements, and open up the possibility for retrospective analysis.


CytoSys: a tool for extracting cell-cycle-related expression dynamics from static data.
Computational models of biological processes are important building blocks in Systems Biology studies. Calibration and validation are two important steps for moving a mathematical model to a computational model. While calibration refers to finding numerical value of the coefficients such as rate constants in a mathematical model, validation refers to verifying that the calibrated model behaves the same as the biological system under previously unseen conditions such as environmental changes (e.g., drug treatment) or mutations. In lieu of direct measurements of rate constants, modeling of the molecular mechanisms that govern biological behaviors may be able to use dynamic expression profiles of reactant biomolecules for calibration. For validation, similar data, obtained under new conditions, are probably better than direct measurements of rate constants. In any case, direct measurement of rate constants is almost always impractical and difficult or impossible. Here, we show a computer-assisted methodology to extract embedded dynamic profiles of cell-cycle proteins from statically sampled, multivariate cytometry data guided by heuristics assembled from canonical cell-cycle knowledge. The methodology is implemented using standard "list mode" cytometry data-processing software followed by CytoSys - a software tool with an easy-to-use graphical interface. We demonstrate the use of CytoSys with a case study of exponentially growing, human erythroleukemia cells and extract the dynamic expression profiles of cyclin A for calibrating an existing deterministic mathematical model of the cell cycle.


Bis-trifluoromethyl effect: doubled transitions in the rotational spectra of hexafluoroisobutene, (CF3)2C═CH2.
Rotational spectra for hexafluoroisobutene, and its (13)C isotopologues, have been recorded between 8 and 16 GHz using a chirped pulse, Fourier transform microwave spectrometer. Notably, all spectra observed are doubled with separations between the doublets being between 1 and 60 MHz. We propose that the bis-trifluoromethyl groups of the target molecule are staggered in the equilibrium configuration, and that a novel, out-of-phase rotation through a F-CCC-F planar configuration with low barrier (<100 cm(-1)), leads to the observed doubled rotational spectra.


Seasonal occurence of infective nematode larvae on irrigated pasture grazed by cattle in California.
During the period extending from Dec 1, 1975, to Dec 22, 1976, 3 distinct peaks in the concentration of infective nematode larvae on pasture forage were detected by use of helminth-free tracer calves. Two of these peaks, 1 occurring in the spring and 1 in the fall, consisted mainly of Ostertagia ostertagi and Cooperia sp. It is concluded that these 2 peaks can be anticipated annually and constitute unsafe periods of grazing. The 3rd peak was comprised largely of Haemonchus placei and Cooperia sp. It is concluded that this peak occurred as the result of unusual rainfall and reduced evaporation in August. Arrested developed (24% to 30%) of O ostertagi larvae occurred only during the spring peak. This finding indicates that on irrigated pastures in California, type I ostertagiasis can be expected to occur as the result of infection during the spring and/or fall peaks, whereas type II ostertagiasis can be expected to occur in the fall as a result of infection obtained during the spring peak. Arrestment in development of Cooperia sp occurred during spring (14%) and fall (23%) peaks. Haemonchus placei larvae increased their propensity for arrestment in development from zero to 82% during the months following the peak level of infection in summer. Other nematodes encountered were Trichostrongylus axei and Nematodirus helvetianus.


Sterilization of infant formula: current practices and recommendations.
Home sterilization of infant formula became a standard of well-baby care in the 1940s. Its purpose was to eliminate bacterial pathogens that could contaminate water and bottles. Public water supplies are now rarely contaminated, and studies have confirmed that sterilization is no longer necessary. Connecticut family physicians and Connecticut mothers were polled and it was found that 48 percent of physicians recommend sterilization and that 54 percent of mothers (using public water supplies) sterilize formula. Formula sterilization recommendations by physicians and sterilization practices by mothers need to be updated.


Pharmacokinetics of high-dose intravenous melatonin in humans.
This crossover study investigated the pharmacokinetics and adverse effects of high-dose intravenous melatonin. Volunteers participated in 3 identical study sessions, receiving an intravenous bolus of 10 mg melatonin, 100 mg melatonin, and placebo. Blood samples were collected at baseline and 0, 60, 120, 180, 240, 300, 360, and 420 minutes after the bolus. Quantitative determination of plasma melatonin concentrations was performed using a radioimmunoassay technique. Pharmacokinetic parameters were estimated by a compartmental pharmacokinetic analysis. Adverse effects included assessments of sedation and registration of other symptoms. Sedation, evaluated as simple reaction times, was measured at baseline and 120, 180, 300, and 420 minutes after the bolus. Twelve male volunteers completed the study. Median (IQR) Cmax after the bolus injections of 10 mg and 100 mg of melatonin were 221,500.0 (185,637.5-326,175.0) pg/mL and 1,251,500.0 (864,375.0-1,770,500.0) pg/mL, respectively; mean (SD) t1/2 was 42.3 (5.6) minutes and 46.2 (6.2) minutes; mean (SD) Vd was 1.6 (0.9) L/kg and 2.0 (0.8) L/kg; mean (SD) CL was 0.0253 (0.0096) L/min · kg and 0.0300 (0.0120) L/min · kg; and median (IQR) AUC0- ∞ , 8,997,633.0 (6,071,696.2-11,602,811.9) pg · min/mL and 54,685,979.4 (36,028,638.6-105,779,612.0) pg · min/mL. High-dose intravenous melatonin did not induce sedation, evaluated as simple reaction times. No adverse effects were reported in the study.


Biology of A beta amyloid in Alzheimer's disease.
The genetic associations with the pathological features of AD are diverse: A rapidly growing number of mutations in presenilin 1 and 2 on chromosomes 14 and 1, respectively, are found in many early-onset FAD patients (Lendon et al., 1997). In addition, beta PP mutations are found in a small percentage of early-onset FAD kindreds. The apoE4 allele on chromosome 19 is associated with the presence of the most common form of AD, sporadic AD (Wisniewski & Frangione, 1992; Namba et al., 1991). However, it is clear that other proteins are also involved in the pathogenesis of AD, since some early-onset FAD kindreds do not have linkage to PS1, PS2, apoE, or beta PP, while at least 50% of late-onset AD is unrelated to apoE. Other proteins which have been implicated in the formation of senile plaques, but so far are not known to have any genetic linkage to AD, include proteoglycans (Snow et al., 1987), apoA1 (Wisniewski et al., 1995a), alpha 1-antichymotrypsin (Abraham et al., 1988), HB-GAM (Wisniewski et al., 1996a), complement components (McGeer & Rogers, 1992), acetylcholinesterase (Friede, 1965), and NAC (Ueda et al., 1993). Which of these proteins will be the most important for the etiology of the most common form of AD, late-onset sporadic AD, remains an open question. Three of the genes which are now known to be linked to AD, including PS1, beta PP, and apoE, have been established immunohistochemically and biochemically to be components of senile plaques (see Fig. 1). This raises at least two possibilities: either each of these proteins is part of one pathway with A beta-related amyloid formation as a final causative pathogenic event or amyloid deposition in AD is a reactive process related to dysfunction of a number of different CNS proteins. Whether or not amyloid formation is directly causative in the pathogenesis of AD, current data suggest that new therapeutic approaches which may inhibit the aggregation and/or the conformational change of sA beta to A beta fibrils (Soto et al., 1996) have the greatest likelihood to make a significant impact on controlling amyloid accumulation in AD.


A 24-Year Follow-up Study on Recidivism in Male Mentally Disordered Sexual Offenders With and Without Psychotic Disorders.
There is a lack of knowledge on mentally disordered sex offenders (MDSOs) targeting adult victims, especially regarding recidivism patterns and the specific subgroup with psychiatric disorders. This paper presents index offense data, clinical data, and recidivism patterns over up to 24 years in a cohort of 146 MDSOs, with and without psychotic disorders, sentenced in Sweden between 1993 and 1997. At the time of the offense, all offenders were affected by clinical, developmental, and criminal history factors. MDSOs with psychotic disorders only marginally differed from those without, the former being less likely to have been institutionalized during childhood, intoxicated during the index offense, or diagnosed with a personality disorder, substance use disorder, or paraphilic disorder. In the cohort, 3.4% of the MDSOs were reconvicted for a new sex offense over 2 years, 9.6% over 5 years, 13.0% over 10 years, and 17.1% over the entire follow-up period of 24 years. In MDSOs with psychotic disorders, no subjects were reconvicted during the first 2 years, while 2.6% were reconvicted over 5 years, 5.3% over 10 years, and 7.9% over 24 years. Recidivism rates for violent and general reoffenses were 39.0% and 37.7%, respectively, for the cohort of MDSOs, and subjects with psychotic disorders reoffended significantly later in general offenses. In conclusion, MDSOs with psychotic disorders showed the same recidivism pattern as MDSOs without psychotic disorders. Furthermore, recidivism research may preferably focus on follow-up periods of 5-10 years since most offenders appear to recidivate within this timeframe.


[A new basic drug prospidin in the treatment of rheumatoid arthritis. III. Comparative analysis of prospidin and cyclophosphamide].
A comparative study of the cytostatic drugs prospidin and cyclophosphamide used in equal doses for rheumatoid arthritis (RA) is reported. Clinical and immunologic effects were determined, and the nature and incidence of side effects and complications were compared. Prospidin showed a more pronounced antirheumatic effect, making for a smaller daily requirement of nonsteroid anti-inflammatory agents or hormones in hormone-dependent cases. Unlike cyclophosphamide, prospidin was not associated with severe side effects and complications precluding further use of the drug. Both drugs demonstrated a regulatory effect on RA-associated immunologic disorders.


Calcium and pH in anoxic and toxic injury.
The critical events that lead to the transition from reversible to irreversible injury remain unclear. Studies are reviewed that have suggested that a rise in cytosolic free Ca2+ initiates plasma membrane bleb formation and a sequence of events that leads ultimately to cell death. In recent studies, we have measured changes in cytosolic free Ca2+, mitochondrial membrane potential, cytosolic pH, and cell surface blebbing in relation to the onset of irreversible injury and cell death following anoxic and toxic injury to single hepatocytes utilizing multiparameter digitized video microscopy (MDVM). MDVM is an emerging new technology that permits single living cells to be labeled with multiple probes whose fluorescence is responsive to specific cellular parameters of interest. Fluorescence images specific for each probe are collected over time, and then digitized and stored. Image analysis and processing then permits quantitation of the spatial distribution of the various parameters within the single living cells. Our results indicate the following: (1) formation of plasma membrane blebs accompanies all types of injury in hepatocytes; (2) cell death is a rapid event, initiated by rupture of a plasma membrane bleb, and is coincident with the onset of irreversible injury; (3) an increase of cytosolic free Ca2+ is not the stimulus for bleb formation or the final common pathway leading to cell death; (4) a decrease of mitochondrial membrane potential precedes loss of cell viability; (5) cytosolic pH falls by more than 1 pH unit during chemical hypoxia. This acidosis protects against the onset of cell death.


The level of provision of specialist palliative care services in Scotland: an international benchmarking study.
Comparative benchmarking of specialist palliative care (SPC) services across jurisdictions can be used to assess the adequacy of provision. Published in 2016, the Scottish Atlas of Palliative Care unlocks the possibility of benchmarking Scotland's provision against other European Union (EU) countries. Our objectives were to describe the provision of SPC services in Scotland and compare this with other EU countries, assessing coverage against European norms. We conducted a secondary analysis of data collected as part for the Scottish Atlas by structured telephone (n=33) or online (n=3) survey with informants from 14 territorial health boards and 15 hospices who provided information about SPC services in their locality. National-level Scottish data were compared with data from other EU countries allowing ranking for each service type and service coverage as calculated against European Association for Palliative Care norms. Scotland had a total of 23 SPC inpatient units containing 349 beds, 27 SPC hospital support teams and 38 SPC home care teams. Relative to other EU countries, Scotland ranked seventh for provision of SPC inpatient units and hospital support teams, and fifth for home care teams. Coverage for these services was 85%, 100% and 72%, respectively. Scotland is positioned among the top 10 EU countries for the level of provision of SPC services. National policy in Scotland has focused on the delivery of palliative care at home or in a homely setting. These data support a focus on developing services in community settings to meet Scotland's policy ambitions.


Continuous Blood Pressure Measurement From Invasive to Unobtrusive: Celebration of 200th Birth Anniversary of Carl Ludwig.
The year 2016 marks the 200th birth anniversary of Carl Friedrich Wilhelm Ludwig (1816-1895). As one of the most remarkable scientists, Ludwig invented the kymograph, which for the first time enabled the recording of continuous blood pressure (BP), opening the door to the modern study of physiology. Almost a century later, intraarterial BP monitoring through an arterial line has been used clinically. Subsequently, arterial tonometry and volume clamp method were developed and applied in continuous BP measurement in a noninvasive way. In the last two decades, additional efforts have been made to transform the method of unobtrusive continuous BP monitoring without the use of a cuff. This review summarizes the key milestones in continuous BP measurement; that is, kymograph, intraarterial BP monitoring, arterial tonometry, volume clamp method, and cuffless BP technologies. Our emphasis is on recent studies of unobtrusive BP measurements as well as on challenges and future directions.


Identifying factors predicting iron deficiency in United States adolescent females using the ferritin and the body iron models.
Iron deficiency is the most prevalent nutritional deficiency in the United States affecting 9-16% of female adolescents. With the primary purpose of detecting iron deficiency, primary care screening consists of a hemoglobin or hematocrit laboratory test. This method is simple and inexpensive, but tests for anemia, and is neither sensitive nor specific for iron deficiency. Alternate methods for diagnosing iron deficiency using the ferritin and body iron models are not widely utilized. The study objective was to compare iron deficiency risk factors among adolescent females defined by the ferritin and body iron models to better characterize those who may benefit from iron deficiency testing as opposed to the current anemia-based screen. This cross-sectional study of female adolescents aged 12-21 years utilized National Health and Nutrition Examination Survey 2003-2006 data. Anemia was defined by standard hemoglobin cutoffs. The ferritin model defines iron deficiency through transferrin saturation, ferritin and erythrocyte protoporphyrin laboratory testing. Body iron calculates iron status with a formula involving transferrin receptor and ferritin. Bivariate and multivariable analyses examined associations between questionnaire responses and iron deficiency defined by each model. Among 1765 participants, 2.7% were anemic. Iron deficiency prevalence was 13.1% and 9.1% by the ferritin and body iron models, respectively. Based on the model, anemia-based screening had a sensitivity of 15.6-18.8% for iron deficiency. Multivariable associations for ferritin model iron deficiency included age, race/ethnicity, activity level and medroxyprogresterone acetate injection. Age and food insecurity were significant using the body iron model. Universal anemia-based screening misses the majority of iron-deficient adolescent females. The common risk factor identified here, adolescent age, may both inform preventive care guidelines on age-based screenings and prospective studies of adolescent iron deficiency risk factors.


Microsolvation effect, hydrogen-bonding pattern, and electron affinity of the uracil-water complexes U-(H2O)n (n = 1, 2, 3).
To achieve a systematic understanding of the influence of microsolvation on the electron accepting behaviors of nucleobases, the reliable theoretical method (B3LYP/DZP++) has been applied to a comprehensive conformational investigation on the uracil-water complexes U-(H(2)O)(n) (n = 1, 2, 3) in both neutral and anionic forms. For the neutral complexes, the conformers of hydration on the O2 of uracil are energetically favored. However, hydration on the O4 atom of uracil is more stable for the radical anions. The electron structure analysis for the H-bonding patterns reveal that the CH...OH(2) type H-bond exists only for di- and trihydrated uracil complexes in which a water dimer or trimer is involved. The electron density structure analysis and the atoms-in-molecules (AIM) analysis for U-(H(2)O)(n) suggest a threshold value of the bond critical point (BCP) density to justify the CH...OH(2) type H-bond; that is, CH...OH(2) could be considered to be a H-bond only when its BCP density value is equal to or larger than 0.010 au. The positive adiabatic electron affinity (AEA) and vertical detachment energy (VDE) values for the uracil-water complexes suggest that these hydrated uracil anions are stable. Moreover, the average AEA and VDE of U-(H(2)O)(n) increase as the number of the hydration waters increases.


Urinary excretion of sulphated N-acetylhexosamines in patients with various mucopolysaccharidoses.
Sulphated N-acetylhexosamines have been isolated from human urine and tentatively identified as N-acetylglucosamine 6-sulphate (GlcNAc6S), N-acetylgalactosamine 6-sulphate (GalNAc6S), N-acetylgalactosamine 4-sulphate (GalNAc4S) and N-acetylgalactosamine 4,6-disulphate (GalNAc4,6diS). Urine from mucopolysaccharidosis-Type-IIID, -IVA and -VI patients compared with that from normal individuals contains elevated levels of GlcNAc6S (380-fold), GalNAc6S (180-fold) and GalNAc4S (420-fold) respectively. Urine from mucopolysaccharidosis-Type-VI patients also contain more than 600 times the normal level of GalNAc4,6diS. Urine from a mucolipidosis-Type-II and a multiple-sulphatase-deficient patient, and, in general, all mucopolysaccharidosis patients studied, contain at least 5-10-fold elevations of sulphated N-acetylhexosamines over the levels detected in urine from normal controls and a alpha-mannosidosis patient. Urine from patients with clinically mild phenotypes contains less sulphated N-acetylhexosamines than isolated from urine of clinically severe mucopolysaccharidosis patients. The source of the four sulphated N-acetylhexosamines is not known. However, incubation of a series of oligosaccharide substrates, derived from keratan sulphate and chondroitin 6-sulphate and containing non-reducing-end beta-linked 6-sulphated N-acetylhexosamine residues, with homogenates of cultured human skin fibroblasts has indirectly been shown to release GlcNA6S and GalNAc6S respectively. Release of GalNAc4S could not be demonstrated in similar incubations of oligosaccharide substrates derived from chondroitin 4-sulphate and containing non-reducing-end beta-linked GalNAc4S residues. We propose that some, if not all, of the sulphated N-acetylhexosamine present in human urine is derived from the action of beta-N-acetylhexosaminidase on sulphated GlcNAc or GalNAc residues at the non-reducing end of keratan sulphate, dermatan sulphate or chondroitin sulphate.


Airway clearance, mucoactive therapies and pulmonary rehabilitation in bronchiectasis.
This paper aims to provide physiological rationale for airway clearance, mucoactive therapy and pulmonary rehabilitation (PR) (or exercise interventions) in bronchiectasis. There is increasing emphasis on the role of airway clearance techniques (ACT) in the management of bronchiectasis. No single ACT has currently shown superior effect over another. Given the large range of different techniques available, consideration of the physiological effects underpinning a technique including expiratory flow, ventilation and oscillation, is essential to effectively personalize ACT. Key clinical trials of mucoactives in bronchiectasis are underway and will provide clarity on the role of these agents in the management of patients with bronchiectasis. Prescription of mucoactive therapies should be done in conjunction with ACT and therefore the mechanism of action of mucoactive drugs and their timing with ACT should be taken into consideration. PR and/or exercise training are recommended in all current bronchiectasis guidelines. There is a clear physiological rationale that muscle weakness and physical inactivity may play a role in disease progression as well as impacting health-related quality of life, frequency of pulmonary exacerbations and ability to mobilize sputum. However, there are residual unanswered questions surrounding the delivery and accessibility to PR. This review summarizes the physiological principles and supporting evidence for airway clearance, mucoactive medication and PR, which are key components in the management of bronchiectasis.


[Pharmaco-angiography of liver tumours (author's transl)].
In 36 patients with clinical suspicion of liver tumours, pharmaco-angiography using adrenalin was added to the usual liver arteriography. In 31 of these patients rumour involvement of the liver was confirmed by surgery, laparoscopy or autopsy. In four out of ten patients with benign liver tumours, pharmaco-angiography lead to improved demonstration and therefore classification of the tumour. Four out of seven malignant primary tumours. Four out of seven malignant primary tumours of the liver were also better demonstrated by pharmaco-angiography. Amongst 14 patients with liver metastases, an improvement in the angiographic demonstration by pharmaco-angiography was achieved in nine. This was particularly the case in tumours showing tumour vessels. Neither benign nor malignant tumours showed constriction of their vessels as a result of pharmaco-angiography and this is therefore not able to differentiate between these. It was also found that tumours showing the the same histology could behave differently during pharmaco-angiography.


Risk factors for elder abuse.
The results of the first national incidence study of noninstitutionalized elder abuse and neglect in the United States are reviewed, as well as underlying causes of abusive relationships involving the elderly. It is estimated that approximately 1% to 2% of elders living in their own homes became abused in the United States during 1996, physically, emotionally, sexually, and/or financially. The abusers were predominantly adult children, spouses, and other relatives. More than 5 times as many new incidents of abuse and neglect were unreported than those reported to authorities responsible for addressing elder abuse. An individual who abuses an elder is often financially dependent on the elder, violent in other contexts, abuses alcohol and/or drugs, and has psychological problems. Although current rules and practices constrain the underwriting professional's use of this information in risk selection, public demand for financial institutions' reporting of elder abuse may provide an opportunity for open discussion about responsible handling of such cases.


Systemic lupus erythematosus and cancer: associated or not?
The frequency of cancer in patients with SLE is between 2.5 and 13.8%. A literature review has identified nine full-length studies that estimated the overall risk of cancer in SLE patients compared with the general population. Five of them have not noted an increased risk for the development of overall cancers among SLE patients compared with the general population. One study identified a 30% increased risk (SIR or 1.3) for occurrence of cancer among 1585 SLE patients followed over 10,807 patient-years. Taken together, it is controversial whether the risk of all cancers is increased in SLE patients compared with the general population. Increased risk of lymphatic malignancies has been shown in multiple large series of SLE patients, but SLE is not associated with an increased risk for the development of most of the solid tumors. Pathogenic mechanisms involved with the development of lymphoproliferative malignancies in association with SLE include a common etiologic agent for both diseases, environmental factors as the use of cytotoxic or immunosuppressive agents, genetic variables, and immunologic factors as immunoregulatory disturbances of the immune system.


Microbial inactivation and evaluation of furan formation in high hydrostatic pressure (HHP) treated vegetable-based infant food.
The inactivation of pathogenic and spoilage bacteria as well as the formation of food processing contaminants (e.g. acrylamide, furan, etc.) in infant foods is of utmost importance for industry, consumers as well as regulatory bodies. In this study, the potential of high hydrostatic pressure (HHP) for microorganism inactivation including total mesophilic aerobic bacteria (TMA) and total yeasts and molds (TYM) at equivalent processing conditions, as well as its effects on furan formation in vegetable-based infant food was evaluated. The process parameters evaluated were combinations of pressures (200, 300, and 400MPa), temperatures (25, 35, and 45°C), and treatment times (5, 10, and 15min). Pressure, time and temperature had a significant influence on both TMA and TYM inactivation of vegetable-based infant foods, observing a significant reduction in both microbial populations when all the factors were increased, although the extent of reduction was clearly influenced by the type of microorganism. A synergism between pressure, time and temperature was observed for the reduction of both TMA and TYM populations and it was found that HHP at 400MPa resulted in a complete inactivation of TMA as well as TYM after 15min of treatment at 45°C. The furan content in all HHP treated samples was found to be below the limit of detection. Thus, HHP treatment could be considered as a potential alternative to thermal processing of vegetable-based infant foods.


Prevalence of IgA-antiendomysial antibody in asymptomatic low bone mineral density.
Celiac disease (CD) is a relatively common gastrointestinal disorder that can be asymptomatic. However, even among asymptomatic patients a long-term reduction in bone mineral density (BMD) is found. Excellent noninvasive screening tests for CD are now available. Studies using older screening techniques have suggested a 10-fold increased prevalence of CD among patients with low BMD, but this has not been confirmed with current testing methodology. We set out to confirm these prevalence estimates using antiendomysial antibody testing. A total of 100 consecutive patients referred to our outpatient endocrinology clinic for evaluation of idiopathic low BMD were studied. In addition to the routine evaluation, patients completed a symptom questionnaire and underwent serological testing for the presence of the IgA antiendomysial antibody (EMA). All patients with a positive EMA underwent small bowel biopsy and permeability studies. EMA results were available on 96 patients; 78/96 patients were female and the mean age was 57 yr (range 18-86 yr). Seven of 96 (7.3% [95% CI 2.1-12.5%]) were EMA-positive, but all tests were low titer (< or = 1:20). However, none of the biopsies showed any histopathological features of CD, nor did EMA status correlate with any of the clinical or laboratory features assessed. Despite a high rate of weakly positive antibody tests, our data do not support an increased prevalence of CD among asymptomatic patients referred for evaluation of low BMD. Without an increase over the background prevalence, the high cost of EMA testing argues against routine use of this test for screening of this population.


Comparison of POCT and central laboratory blood glucose results using arterial, capillary, and venous samples from MICU patients on a tight glycemic protocol.
Point of care (POC) glucose meters are routinely used to monitor glucose levels for patients on tight glycemic control therapy. We determined if glucose values were different for a POC glucose meter as compared to the main clinical laboratory for medical intensive care unit patients on a tight glycemic protocol and whether the site of blood sampling had a significant impact on glucose values. Eighty-four patients (114 paired samples) who were on a tight glycemic protocol in the period November 2005 through August 2006 were enrolled. After simultaneous blood draws, we compared the glucose levels for the glucose meter (arterial/venous/capillary), blood gas (arterial/venous), and central clinical laboratory (serum/plasma from arterial/venous samples). The mean glucose levels of all arterial/venous/fingerstick samples using the glucose meter demonstrated a positive bias of 0.7-0.9 mmol/l (12.6-16.2 mg/dl) (p<0.001) relative to central laboratory venous plasma. There was also a smaller positive (0.1-0.3 mmol/l or 1.8-5.4 mg/dl, p<0.05) bias for arterial/venous blood gas samples and laboratory arterial serum/plasma glucose samples. Using Parkes error grid analysis we were able to show that the bias for arterial or venous POC glucose results would have not impacted clinical care. This was not the case, however, for fingerstick sampling where a high bias could have significantly impacted clinical care. Additionally, in 3 fingerstick samples a severe underestimation (<46% of the central laboratory plasma result) was found. Glucose meters using arterial/venous whole blood may be utilized in the MICU; however, due to the increased variability of results we do not recommend the routine use of capillary blood sampling for monitoring glucose levels in the MICU setting.


Evaluating the existence and benefit of major histocompatibility complex-based mate choice in an isolated owl population.
How mate preferences evolve in the first place has been a major conundrum for sexual selection. Some hypotheses explaining this assume fitness benefit derived from subsequent generations. Major histocompatibility complex (MHC)-based mate choice is a representative example of the mate choice that is associated with such trans-generational mechanisms. To provide evidences for fitness benefit of MHC-based mate choice, previous studies assessed the association between own MHC genotype and own fitness components. However, the association between MHC-based mate choice in the parental generation and fitness components in the resultant offspring generation has only rarely been measured in wild populations. Focusing on the isolated population of the monogamous Ryukyu Scops Owl (Otus elegans interpositus) on Minami-daito Island, Japan, we found evidence of MHC-based mate choice. However, we found no evidence of MHC-based mate choice increasing own reproductive success or offspring survival. This is a rare case study that directly examines the existence of the trans-generational indirect benefit of MHC-based mate choice for genetic compatibility from trans-generational data in a wild bird population. By investigating the fitness benefits of mate choice, this study serves to facilitate our understanding of the evolution of MHC-based mate choice.


Multiresolution feature detection using a family of isotropic bandpass filters.
Presents a novel method which uses a special type of multi-scale isotropic band-pass filters to detect image landmark and corner features. This paper mainly contributes to the solutions of the following problems: (1) defining a general family of feature detectors under a unified framework that are able to enhance and detect the desired features; (2) explaining theoretically and experimentally why these feature points can be detected by the proposed detectors; and (3) extending the detectors to multi-scale versions for jointly achieving good detectability and localization where an automatic scale selection method is applied. The paper then presents several applications for detecting landmarks and corners using the proposed methods, in order to illustrate their usage. These include detecting landmarks from gesture images (of the face and hand), from airborne and vehicle-borne IR landmine images, and from images containing object corners. Experiments have been performed using the proposed detectors to these applications. Some comparisons and evaluations have also been performed. The results have demonstrated the effectiveness of the proposed detectors in terms of feature detectability and localization.


[Comparison of susceptibility artefacts of different radiofrequency electrodes at O.2 T. Influence of electrode positioning, pulse sequence and image reconstruction methods].
Interventional MRI procedure monitoring requires small but accurate susceptibility artifacts of the instruments used. In this investigation, susceptibility artifacts of different RF-electrode designs were compared using a variety of pulse sequences and k-space acquisition methods. 4 different 18-gauge RF-electrodes (with three single electrodes made of stainless steel, copper, inconal, and a triple-clustered electrode configuration made of inconal) were placed in a 0.2 T MR-scanner perpendicular to the main magnetic field. Pulse sequences used included: TSE T2, FISP, true-FISP, PSIF, and a temperature sensitive ES-GRE sequence. In addition to the 2D Cartesian k-space trajectory with Fourier transformation (2DFT), projection reconstruction (PR) was used with the FISP, true-FISP and PSIF sequences. The best tip accuracy was achieved with the combination of inconal electrodes and TSE T2. The usefulness of the tested sequences was found to be: TSE T2 > PSIF > FISP/true-FISP > ES-GRE. In general 2DFT provided better or equal tip accuracy than PR. The apparent shaft width was smaller using the copper electrode compared to the inconal electrode. However, the "match shaped" tip artifact of the copper probe led to a higher error in tip accuracy. TSE-T2 sequences and Cartesian 2DFT acquisitions should be used for accurate tip positioning at 0.2 T. Further, artifact size of the electrode shaft prevents the use of inconal for temperature sensitive sequences. Copper electrodes can be used for these purposes, although copper is not considered to be biocompatible at present.


Proteasome inhibitor bortezomib suppresses nuclear factor-kappa B activation and ameliorates eye inflammation in experimental autoimmune uveitis.
Bortezomib is a proteasome inhibitor used for hematologic cancer treatment. Since it can suppress NF-κB activation, which is critical for the inflammatory process, bortezomib has been found to possess anti-inflammatory activity. In this study, we evaluated the effect of bortezomib on experimental autoimmune uveitis (EAU) in mice and investigated the potential mechanisms related to NF-κB inactivation. High-dose bortezomib (0.75 mg/kg), low-dose bortezomib (0.15 mg/kg), or phosphate buffered saline was given after EAU induction. We found that the EAU is ameliorated by high-dose bortezomib treatment when compared with low-dose bortezomib or PBS treatment. The DNA-binding activity of NF-κB was suppressed and expression of several key inflammatory mediators including TNF-α, IL-1α, IL-1β, IL-12, IL-17, and MCP-1 was lowered in the high-dose bortezomib-treated group. These results suggest that proteasome inhibition is a promising treatment strategy for autoimmune uveitis.


Total knee arthroplasty in patient with paraplegia after spinal cord injury.
The clinical management of paraplegic patients is more complex than in able-bodied subjects. Spinal cord injury (SCI) affects younger, active people more often than the elderly during high-energy fall or traffic accidents. In order to return to work after suffering an SCI, patients need to regain their functional independence, especially their ability to drive. The literature lacks strong evidence addressing the surgical solutions in severe knee arthrosis in paralyzed patients after SCI. We present a favourable outcome of total knee arthroplasty (TKA) of a stiff knee in extension in a man with T12 grade C paraplegia after SCI. We describe an effective rehabilitation protocol after knee arthroplasty in patient with damage to the spinal cord. Several factors should be taken into account before performing surgery: 1. ability of regaining some of spinal cord locomotor function through intensive gait rehabilitation in SCI patients, 2. presence of muscle imbalance and knee contractures combined with a risk of bone fracture resulting from intensive postoperative rehabilitation, 3. the impaired microvasculature of the skin and subcutaneous tissues and increased risk of occlusion occurrence of the capillaries and small vessels of the leg, 4. higher prevalence of secondary infections via urinary entry sites in patients after SCI, 5. patient's strong determination and willingness to undergo the arthroplasty procedure. TKA might be considered in selected paralyzed patients after SCI, especially in those with severe arthrosis as well as significant knee contractures. Our study reveals the advantage of performing TKA in improving functional state in patients with cord injury.


Thyroid and Islet Autoantibodies Predict Autoimmune Thyroid Disease at Type 1 Diabetes Diagnosis.
Screening of autoimmune thyroid disease in children with type 1 diabetes is important but varies between clinics. To determine the predictive value of thyroid autoantibodies, thyroid function, islet autoantibodies, and HLA-DQ at diagnosis of type 1 diabetes for autoimmune thyroid disease during follow-up. Forty-three Swedish pediatric endocrinology units. At diagnosis of type 1 diabetes, autoantibodies against thyroid peroxidase (TPOAb), thyroglobulin (TGAb), glutamic acid decarboxylase (GADA), insulin, insulinoma-associated protein-2, and 3 variants of zinc transporter 8 (ZnT8W/R/QA) HLA-DQA1-B1 genotypes and thyroid function were analyzed in 2433 children. After 5.1 to 9.5 years, information on thyroxine treatment was gathered from the Swedish National Board of Health and Welfare's Prescribed Drug Register. Thyroxine was prescribed to 6% of patients. In patients <5 years of age, female sex [hazard ratio (HR) = 4.60; P = 0.008] and GADA (HR = 5.80; P = 0.02) were predictors. In patients 5 to 10 years old, TPOAb (HR = 20.56; P < 0.0001), TGAb (HR = 3.40; P = 0.006), and thyroid-stimulating hormone (TSH) (HR = 3.64; P < 0.001) were predictors, whereas in 10 to 15 year olds, TPOAb (HR = 17.00; P < 0.001) and TSH (HR = 4.11; P < 0.001) predicted thyroxine prescription. In addition to TPOAb and TSH, GADA at diagnosis of type 1 diabetes is important for the prediction of autoimmune thyroid disease in children <5 years of age.


The ultrastructure of Cynodon salt glands: the apoplast.
The salt glands of Bermuda grass were found to be similar to those of many dicotyledonous plants with respect to the relatively large quantities of chloride that accumulate in the subcuticular collecting chamber of secreting glands. The salt glands of grasses lack the suberized or cuticularized zone that is present in the walls of the glands of dicotyledonous plants that has been hypothesized to prevent the apoplastic backflow of accumulated salts from the collecting chamber to the leaf mesophyll. An ionic lanthanum solution was used to determine whether or not the apoplastic pathway is blocked between the collecting chamber and the leaf mesophyll. The results of the lanthanum tracer study revealed that this pathway is significantly restricted. It was determined that the restriction of the apoplastic pathway was due to the highly lignified condition of the lateral cell walls in the outer portion of the basal gland cell. The lanthanum tracer study also revealed that an apoplastic continuum exists between the leaf mesophyll and a system of membranous extracellular channels that occur in the basal cell. Based on this finding, it was suggested that the extracellular channels may function in the absorption of solutes from the apoplast.


LSD1/KDM1A is essential for neural stem cell differentiation in mice.
The proper regulation of neural stem cell differentiation is required for the proper specification of the central nervous system. Here we investigated the function of the H3K4me1/2 demethylase LSD1/KDM1A during neural stem differentiation in mice. Conditional deletion of LSD1 in nestin - positive neural stem cells results in 100% perinatal lethality after birth with severe motor coordination deficits, retarded growth and defects in brain morphology. Despite these severe defects, motor neuron progenitors and the initial motor neuron population are specified normally and motor neurons with normal morphology can be cultured from these mice in vitro . However, motor neurons cultured from mice lacking LSD1 in neural stem cells continue to inappropriately maintain critical neural stem cell proteins. Taken together these results suggest that, as in other mouse stem cell populations, LSD1 is required to deactivate the stem cell program to enable normal neural stem cell differentiation. However, unlike in other mouse stem cell populations, the inappropriate maintenance of the stem cell program during neural stem cell differentiation may compromise neuronal function rather than neuronal specification.


[Experimental, morphological and clinical aspects of transmyocardial laser revascularization].
Experimental, pathomorphological and clinical data substantiating use of transmyocardial laser revascularisation in the treatment of coronary heart disease (CHD) are presented. It is shown that the objective criterion of clinical efficacy of transmyocardial laser revascularisation in 58 patients with CHD is an increase of myocardial perfusion due to neoangiogenesis. This leads to improvement of cardiac function and general state of the patients. Surgical intervention has no substantial influence on contractile function of left ventricular myocardium in patients with CHD.


The EMAPII cytokine is released from the mammalian multisynthetase complex after cleavage of its p43/proEMAPII component.
Endothelial-monocyte-activating polypeptide II (EMAPII) is an inflammatory cytokine released under apoptotic conditions. Its proEMAPII precursor proved to be identical to the auxiliary p43 component of the aminoacyl-tRNA synthetase complex. We show here that the EMAPII domain of p43 is released readily from the complex after in vitro digestion with caspase 7 and is able to induce migration of human mononuclear phagocytes. The N terminus of in vitro-processed EMAPII coincides exactly with that of the mature cytokine isolated from conditioned medium of fibrosarcoma cells. We also show that p43/proEMAPII has a strong tRNA binding capacity (K(D) = 0.2 microm) as compared with its isolated N or C domains (7.5 microm and 40 microm, respectively). The potent general RNA binding capacity ascribed to p43/proEMAPII is lost upon the release of the EMAPII domain. This suggests that after onset of apoptosis, the first consequence of the cleavage of p43 is to limit the availability of tRNA for aminoacyl-tRNA synthetases associated within the complex. Translation arrest is accompanied by the release of the EMAPII cytokine that plays a role in the engulfment of apoptotic cells by attracting phagocytes. As a consequence, p43 compares well with a molecular fuse that triggers the irreversible cell growth/cell death transition induced under apoptotic conditions.


Organic light emitting diode improves diabetic cutaneous wound healing in rats.
A major complication for diabetic patients is chronic wounds due to impaired wound healing. It is well documented that visible red wavelengths can accelerate wound healing in diabetic animal models and patients. In vitro and in vivo diabetic models were used to investigate the effects of organic light emitting diode (OLED) irradiation on cellular function and cutaneous wound healing. Human dermal fibroblasts were cultured in hyperglycemic medium (glucose concentration 180 mM) and irradiated with an OLED (623 nm wavelength peak, range from 560 to 770 nm, power density 7 or 10 mW/cm2 at 0.2, 1, or 5 J/cm2). The OLED significantly increased total adenosine triphosphate concentration, metabolic activity, and cell proliferation compared with untreated controls in most parameters tested. For the in vivo experiment, OLED and laser (635 ± 5 nm wavelength) treatments (10 mW/cm2 , 5 J/cm2 daily for a total of seven consecutive days) for cutaneous wound healing were compared using a genetic, diabetic rat model. Both treatments had significantly higher percentage of wound closure on day 6 postinjury and higher total histological scores on day 13 postinjury compared with control. No statistical difference was found between the two treatments. OLED irradiation significantly increased fibroblast growth factor-2 expression at 36-hour postinjury and enhanced macrophage activation during initial stages of wound healing. In conclusion, the OLED and laser had comparative effects on enhancing diabetic wound healing.


In vitro activities of various antimicrobials alone and in combination with tigecycline against carbapenem-intermediate or -resistant Acinetobacter baumannii.
The activities of tigecycline alone and in combination with other antimicrobials are not well defined for carbapenem-intermediate or -resistant Acinetobacter baumannii (CIRA). Pharmacodynamic activity is even less well defined when clinically achievable serum concentrations are considered. Antimicrobial susceptibility testing of clinical CIRA isolates from 2001 to 2005 was performed by broth or agar dilution, as appropriate. Tigecycline concentrations were serially increased in time-kill studies with a representative of the most prevalent carbapenem-resistant clone (strain AA557; imipenem MIC, 64 mg/liter). The in vitro susceptibility of the strain was tested by time-kill studies in duplicate against the average free serum steady-state concentrations of tigecycline alone and in combination with various antimicrobials. Ninety-three CIRA isolates were tested and were found to have the following antimicrobial susceptibility profiles: tigecycline, MIC(50) of 1 mg/liter and MIC(90) of 2 mg/liter; minocycline, MIC(50) of 0.5 mg/liter and MIC(90) of 8 mg/liter; doxycycline, MIC(50) of 2 mg/liter and MIC(90) of > or =32 mg/liter; ampicillin-sulbactam, MIC(50) of 48 mg/liter and MIC(90) of 96 mg/liter; ciprofloxacin, MIC(50) of > or =16 mg/liter and MIC(90) of > or =16 mg/liter; rifampin, MIC(50) of 4 mg/liter and MIC(90) of 8 mg/liter; polymyxin B, MIC(50) of 1 mg/liter and MIC(90) of 1 mg/liter; amikacin, MIC(50) of 32 mg/liter and MIC(90) of > or =32 mg/liter; meropenem, MIC(50) of 16 mg/liter and MIC(90) of > or =128 mg/liter; and imipenem, MIC(50) of 4 mg/liter and MIC(90) of 64 mg/liter. Among the tetracyclines, the isolates were more susceptible to tigecycline than minocycline and doxycycline, according to FDA breakpoints (95%, 88%, and 71% of the isolates were susceptible to tigecycline, minocycline, and doxycycline, respectively). Concentration escalation studies with tigecycline revealed a maximal killing effect near the MIC, with no additional extent or rate of killing at concentrations 2x to 4x the MIC for tigecycline. Time-kill studies demonstrated indifference for tigecycline in combination with the antimicrobials tested. Polymyxin B, minocycline, and tigecycline are the most active antimicrobials in vitro against CIRA. Concentration escalation studies demonstrate that tigecycline may need to approach concentrations higher than those currently achieved in the bloodstream to adequately treat CIRA bloodstream infections. Future studies should evaluate these findings in vivo.


Incomplete contact site A glycoprotein in HL220, a modB mutant of Dictyostelium discoideum.
HL220, a modB mutant that lacks a modification of certain membrane proteins of Dictyostelium discoideum, has been shown to aggregate and to form EDTA-stable intercellular contacts typical of aggregating wild-type cells. A developmentally regulated glycoprotein of 80 X 10(3) apparent molecular weight has been identified as a target site of adhesion-blocking Fab and thought to be involved in EDTA-stable cell contact formation (Müller & Gerisch, 1978). In the HL220 mutant this glycoprotein is no longer recognized by a modB-specific antibody. Therefore, it has been suggested that the 80 X 10(3) Mr glycoprotein, or a modification on it, is not required for the EDTA-stable cell contact of aggregating cells. We show that HL220 synthesizes an equivalent of the 80 X 10(3) Mr glycoprotein with an apparent molecular weight of 68 X 10(3). The mutant product reacted with certain monoclonal antibodies highly specific for the 80 X 10(3) Mr glycoprotein in the wild type, and was developmentally regulated like the 80 X 10(3) Mr glycoprotein. These results indicate that the 68 X 10(3) Mr protein of the mutant lacks a modification, most likely an oligosaccharide residue, the absence of which causes the substantial shift of the apparent molecular weight from 80 X 10(3) to 68 X 10(3). Monoclonal antibodies that did not react with proteins of the mutant could be classified according to their reactions with different sub-sets of wild-type proteins. These results indicate that the proteins that reacted with either one or the other antibody were not modified by a uniform structure. The modification rather varies from one sub-set of cross-reacting proteins to another, suggesting differences between the glycosyl residues of the partially cross-reacting proteins. HL220 cells showed strongly reduced EDTA-stable contact formation under our conditions. EDTA-sensitive intercellular adhesion was undetectable in the mutant, whereas adhesion of the cells to the substratum appeared to be strengthened. The rear ends of the cells, in particular, were tightly attached to glass or Teflon surfaces. The mutant cells were capable of responding chemotactically. Propagated excitation waves like those known to be based on periodic cyclic AMP production and relay were clearly seen. Extracellular phosphodiesterase induction by cyclic AMP and phosphodiesterase inhibitor production were normal. These results indicate that the generation of chemotactic signals and the cellular responses to cyclic AMP are not severely affected by the mutation.


Combinatorial approaches for developing upconverting nanomaterials: high-throughput screening, modeling, and applications.
Colloidal nanoparticles doped with lanthanide ions can upconvert near-infrared light to visible frequencies, enabling the application of such materials to biological imaging and luminescent solar concentration. The optical properties of upconverting nanomaterials are determined by their combination of lanthanide dopants, by their morphology, by their host matrices, and by their surface ligands. Identifying ideal compositions and synthesis conditions for these materials can be tedious and time-consuming due to the large number of parameters to optimize. This review surveys the use of combinatorial strategies to rapidly screen and optimize diverse libraries of upconverting nanomaterials. I will review high-throughput techniques for synthesizing and characterizing large libraries of nanocrystals, and I will discuss theoretical methods for modeling the optical properties of lanthanide-doped materials. Case studies will illustrate the use of these approaches for optimizing the physical properties of upconverting nanoparticles, including cases in which unexpected phenomena were revealed. Finally, this review will identify promising opportunities in which combinatorial techniques could accelerate on-going research or facilitate the discovery of novel upconverting nanomaterials that overcome fundamental limitations of current material designs.


Chemoreceptor reflexes and cardiovascular control.
Evidence is presented which indicates that in the absence of other known inputs to the nervous system and during controlled pulmonary ventilation, stimulation of the carotid body chemoreceptors causes bradycardia and selective peripheral vasoconstriction. These responses may be attenuated, however, by concomitant changes in respiration and arterial blood pressure, and by activity of higher parts of the brain stem. Stimulation of the aortic bodies in mammals in which they are functionally active, causes bradycardia or tachycardia and selective peripheral vasoconstriction. The reflex vascular effects from the peripheral arterial chemoreceptors are mediated by alpha-adrenergic sympathetic fibres. A potential mechanism exists therefore whereby the peripheral arterial chemoreceptors could contribute to the neurogenic component of hypertension.


[Natural antibodies (isohemagglutinins, heterophilic, antierythrocytic and antibacterial) in children with congenital immunologic deficiency syndromes].
Examination of 30 patients with immunodeficient diseases showed that the system of natural antibodies was considerably changed depending on the form and the extent of deficiency of the immune system: in agammaglobuinemia the antibodies under study were almost completely absent, and in immunological insufficiency with ataxia-teleangiectasis the production of antibacterial antibodies proved to be sharply decreased. The data obtained can be used both for the diagnosis of immunodeficiencies and for control of the restoration of the immunological competence after a number of therapeutic measures, particularly after the transplantation of the thymus-sternum complex.


Multilevel statistical inference from functional near-infrared spectroscopy data during stroop interference.
Functional near-infrared spectroscopy (fNIRS) is an emerging technique for monitoring the concentration changes of oxy- and deoxy-hemoglobin (oxy-Hb and deoxy-Hb) in the brain. An important consideration in fNIRS-based neuroimaging modality is to conduct group-level analysis from a set of time series measured from a group of subjects. We investigate the feasibility of multilevel statistical inference for fNIRS. As a case study, we search for hemodynamic activations in the prefrontal cortex during Stroop interference. Hierarchical general linear model (GLM) is used for making this multilevel analysis. Activation patterns both at the subject and group level are investigated on a comparative basis using various classical and Bayesian inference methods. All methods showed consistent left lateral prefrontal cortex activation for oxy-Hb during interference condition, while the effects were much less pronounced for deoxy-Hb. Our analysis showed that mixed effects or Bayesian models are more convenient for faithful analysis of fNIRS data. We arrived at two important conclusions. First, fNIRS has the capability to identify activations at the group level, and second, the mixed effects or Bayesian model is the appropriate mechanism to pass from subject to group-level inference.


Basolateral membrane properties in proximal convoluted tubules of the newborn rabbit.
We examined the properties of basolateral cell membranes in "immature" and "mature" proximal tubules and evaluated the possibility that changes in those membranes might contribute to the reported maturation of tubule function. Proximal convoluted tubules were dissected in connection with superficial glomeruli, crimped at both ends, and induced to swell by use of ouabain, low protein bathing medium, and/or collagenase. Tubules from adult animals swelled faster than tubules from 2- to 5- or 14- to 17-day-old animals. However, the basolateral cell membranes of tubules from adult animals were three- to fourfold larger in surface area than those from 2- to 17-day animals, so that the ratio of the swelling rates to the measured basolateral membrane areas remained constant during the period from newborn to adult. We conclude that the basolateral membranes "mature" only in their surface areas and that little change occurs in their intrinsic properties such as hydraulic conductivity or ion permeability.


Early prediction of transition to durable mechanical circulatory support in patients undergoing peripheral veno-arterial extracorporeal membrane oxygenation for critical cardiogenic shock.
Peripheral veno-arterial extracorporeal membrane oxygenation (pVA-ECMO) has gained increasing value in the management of patients with critical cardiogenic shock (cCS), allowing time for myocardial recovery. Failure of myocardial recovery has life-altering consequences: transition to durable mechanical circulatory support (dMCS), urgent heart transplantation, or withdrawal of support. Clinical factors controlling myocardial recovery under these circumstances remain largely unknown. Using a retrospective cohort, we developed a model for early prediction of transition to dMCS in patients undergoing pVA-ECMO for cCS. To promote myocardial recovery, our clinical management centered around left ventricular pressure unloading, that is, targeting pulmonary capillary wedge pressures (PCWP) ≤18 mm Hg. We collected demographic data, laboratory findings, inotrope use, and two-dimensional transthoracic echocardiography measurements, all limited to the first 72h of pVA-ECMO (D1-3). Out of 70 patients who were alive after pVA-ECMO, 27 patients underwent implantation of dMCS. There was no significant difference in survival to hospital discharge between patients with or without transition to dMCS. Ejection fractionD1-3 (per 10% increase, OR 0.37 [0.17-0.79]) and amount of inotropic supportD1-3 (OR 4.77 [1.6-14.18]) but neither myocardial wall tension nor PCWP emerged as significant predictors of transition to dMCS. Optimism-corrected c-index (0.90 [0.89-0.90]) revealed an excellent discriminative ability of our model. In summary, our model for early prediction of transition to dMCS in patients with cCS undergoing pVA-ECMO identifies indicators of inotropic state as relevant factors. Absence of markers for myocardial oxygen consumption or left ventricular pressure loading allows us to hypothesize sufficient cardiac unloading in our cohort with PCWP-targeted management.


Dimensional crossover and cold-atom realization of topological Mott insulators.
Interacting cold-atomic gases in optical lattices offer an experimental approach to outstanding problems of many body physics. One important example is the interplay of interaction and topology which promises to generate a variety of exotic phases such as the fractionalized Chern insulator or the topological Mott insulator. Both theoretically understanding these states of matter and finding suitable systems that host them have proven to be challenging problems. Here we propose a cold-atom setup where Hubbard on-site interactions give rise to spin liquid-like phases: weak and strong topological Mott insulators. They represent the celebrated paradigm of an interacting and topological quantum state with fractionalized spinon excitations that inherit the topology of the non-interacting system. Our proposal shall help to pave the way for a controlled experimental investigation of this exotic state of matter in optical lattices. Furthermore, it allows for the investigation of a dimensional crossover from a two-dimensional quantum spin Hall insulating phase to a three-dimensional strong topological insulator by tuning the hopping between the layers.


Early Nutrition Intervention Attenuates Weight Gain for Pediatric Acute Lymphoblastic Leukemia Patients in Maintenance Therapy.
Obesity following treatment of pediatric acute lymphoblastic leukemia (ALL) has become a significant long-term concern. Excessive weight gain often occurs during treatment, particularly during induction and the first 6 months of maintenance therapy, and it may be potentially modifiable. This retrospective study aimed to evaluate the impact of an early, 3-visit nutrition intervention on weight gain during maintenance therapy in ALL patients. Medical records of the intervention group were compared with historical controls who were treated on the same ALL treatment protocols during an earlier time period. Anthropometrics were collected throughout intensive therapy and at every monthly visit during the first 12 months of maintenance therapy. In total, 67 patients were evaluated (33 in the intervention group and 34 in the control group). After controlling for significant predictors of body mass index (BMI) z-scores in maintenance therapy-including higher BMI at diagnosis and weight gain throughout intensive therapy-the intervention group demonstrated more controlled weight gain during maintenance therapy (P<0.0001). A 3-visit nutrition intervention was effective in attenuating weight gain trends during ALL maintenance therapy.


Primary yolk sac tumour of the liver in adulthood.
Primary yolk sac tumour of the liver is exceedingly rare. A 28 year old woman presented with a cystic liver mass and a markedly raised serum alpha-fetoprotein concentration. She underwent a partial hepatectomy for a suspected hepatocellular carcinoma but histological examination of the tumour revealed the classical morphological and immunohistochemical features of a yolk sac tumour. There was no evidence of an extrahepatic primary source. Review of this case, together with the six previously reported adult cases of primary yolk sac tumours of the liver, revealed several features of the tumour that may aid differentiation from hepatocellular carcinoma, with potential therapeutic implications.


A GOG 210 aCGH study of gain at 1q23 in endometrioid endometrial cancer in the context of racial disparity and outcome.
The goal of this study was to identify recurrent regions of genomic gain or loss in endometrial cancer of the endometrioid type in the context of racial disparities in mortality for this disease. Array comparative genomic hybridization (aCGH) analysis was performed on 80 frozen primary tumors from the Gynecologic Oncology Group (GOG)-210 bank using the RPCI 19K BAC arrays. The 80 patients included 20 African American (AA) Stage I, 20 White (W) Stage I, 20 African American (AA) Stage IIIC/IV, and 20 White (W) Stage IIIC/IV. A separate subset of 220 endometrial cancers with outcome data was used for validation. A 1.6-Mbp region of gain at 1q23 was identified by aCGH in all AA patients and high grade W patients, but not W low grade patients. In the validation arm of 220 patients copy number gain at this region was validated using FISH and locus specific BACs. The number of AA patients in the validation arm was too small to confirm the aCGH association with racial disparity. Kaplan-Meier curves for survival showed a significant difference for gain at 1q23 versus no gain (log rank P = 0.0014). When subdivided into various groups of risk by stage and grade the survival curves showed a decreased survival for high grade and/or stage tumors, but not for low grade and/or stage endometrioid tumors. Univariate analyses for gain at 1q23 showed a significant association (P = 0.009) with survival. Multivariate analysis for gain at 1q23 did not show a significant association with survival (P = 0.14).


Molecular cloning, expression and exon/intron organization of the bovine beta-galactoside alpha2,6-sialyltransferase gene.
In this study, we report the first isolation and characterization of a bovine sialyltransferase gene. Bovine cDNAs prepared from different tissues contain an open-reading frame encoding a 405 amino acid sequence showing 83%, 75%, and 60% identity with human, murine, and chicken ST6Gal I (beta-galactoside alpha2,6-sialyltransferase) sequences, respectively. When transfected into COS-7 cells, a recombinant enzyme was obtained which catalyzed the in vitro alpha2, 6-sialylation of LacNAc (NeuAcalpha2-6Galbeta1-4GlcNAc) and LacdiNAc (NeuAcalpha2-6GalNAcbeta1-4GlcNAc) acceptor substrates. The K (m) values were 2.8 and 6.9 mM, respectively. Different relative efficiencies (Vmax/Km) for the two precursors (36 for LacNAc and 4.3 for LacdiNAc) were observed. Bovine ST6Gal I gene consists of four 5'-untranslated exons E(-2) to E(1), and five coding exons from E(2) to E(6). This later carries a 3'-untranslated region of 2. 7 kb. Gene sequence spans at least 80 kb of genomic DNA. Two processed pseudogenes have been identified. They are 94.3 and 95.6% similar to the bovine cDNA, respectively. Three families of mRNA isoforms were isolated. They differed by their 5'-untranslated regions and could be generated by three tissue-specific promoters. Family 1 is made up of exons E(-2) and E(1) to E(6), family 2 of exons E(-1) to E(6), and family 3 of exons E(1) to E(6). Tissular distribution of transcript families appears noticeably different than those described in human and rat.


Environmental stimulation reduces learning deficits in experimental cretinism.
Behavioral deficits in adult rats exposed perinatally to thiouracil were substantially reduced or elimated by a 5-week period of "superenriched" postweaning rearing conditions before testing. This treatment resulted in remediation of hypothyroid rats' deficits in maze learning, maze retention, and resistance to extinction of bar-pressing; the facilitative effect persisted for more than 4 months. These behavioral results were consistent with neurohistological findings from studies of early thyroid deficiency and postweaning environmental stimulation in rats.


Lung tumor cells inhibit bone mineralization and osteoblast activity.
Patients with non-small cell lung cancer (NSLC) often develop skeletal complications and fractures. To understand mechanisms of bone loss, we developed a murine model of non-metastatic NSLC. Decreased bone mineral density, trabecular thickness and mineralization, without an increase in bone resorption, were observed in vivo in mice injected with Lewis lung adenocarcinoma (LLC1) cells in the absence of tumor cell metastases. A decrease in trabecular bone mineral density was observed in mice injected with cell-free LLC1 CM. Plasma osteoblast biomarkers and PTH-related peptide (PTHrP) were reduced, and parathyroid hormone (PTH), 1,25-dihydroxyvitamin D, calcium and phosphate concentrations were normal in tumor-bearing mice. LLC1 cell conditioned medium (CM) inhibited alkaline phosphatase activity, osteoblast mineralization, and expression of Alpl and Ocn/Bglap mRNA in MC3T3 osteoblast cultures, whereas non-CM or CM from NIH/3T3 fibroblasts did not induce similar changes. LLC1 CM reduced Wnt3a-stimulated Tcf/Lef reporter plasmid activity and Wnt5A, Tcf1 and Lef1 mRNA expression in MC3T3 cells. Although concentrations of the Wnt inhibitor, DKK2, were increased in LLC1 CM compared to non-CM, depletion of DKK2 from LLC1 CM did not completely restore Wnt3a activity in MC3T3 cultures, and recombinant DKK2 failed to inhibit osteoblast mineralization. The data indicate that in a model of lung adenocarcinoma without bone metastases, tumor cells elaborate a secreted factor(s) that reduces bone mass, bone formation and osteoblast Wnt signaling without increases in bone resorption or calcium-regulating hormone concentrations. The factor(s) mediating this inhibition of osteoblast mineralization require further characterization.


Multidisciplinary guidelines for initial evaluation of complicated lymphatic anomalies-expert opinion consensus.
Complicated lymphatic anomalies (CLAs) are chronic, progressive, and debilitating conditions that share clinical features, yet key elements for optimal evaluation and management have not been established. We aimed to formulate expert opinion consensus-based guidelines for comprehensive evaluation of CLAs. Patient support groups dedicated to CLAs organized an international conference for vascular anomaly experts from 16 specialties to address the objective. Participants received a set of questions before the meeting and reviewed the literature. Data extracted from international lymphatic anomaly registries were presented and the group separated for panel discussions during the conference. The recommendations achieving consensus within the panel were presented to the entire audience. Open debate occurred until majority approval was achieved. The expert group was composed of 52 physicians who defined the clinical elements required to evaluate and diagnose a CLA. The radiology panel established the preferred anatomical and functional imaging methods for diagnosis and the elements required to be described during interpretation. Two medical panels compiled the metabolic and hematologic tests at diagnosis and also recommended functional studies. The surgical group recommended precautions for biopsy and the pathology panel provided biopsy specimen processing guidelines. Patients with CLAs require a comprehensive and targeted diagnostic plan for appropriate management, prevention of complications, and conservation of resources. As this population is managed by diverse medical and surgical specialties, we offer an expert multidisciplinary consensus-based opinion on the current literature and on data extracted from international lymphatic anomaly registries.


Human liver aldehyde reductase: pH dependence of steady-state kinetic parameters.
The pH dependence of steady-state parameters for aldehyde reduction and alcohol oxidation were determined in the human liver aldehyde reductase reaction. The maximum velocity of aldehyde reduction with NADPH or 3-acetyl pyridine adenine dinucleotide phosphate (3-APADPH) was pH independent at low pH but decreased at high pH with a pK of 8.9-9.6. The V/K for both nucleotides decreased below a pK of 5.7-6.2, as did the pKi of competitive inhibitors NADP and ATP-ribose, suggesting that the 2'-phosphate of the nucleotide has to be deprotonated for binding to the enzyme. The pK of the 2'-phosphate of NADPH appears to be perturbed in the ternary complexes to 5.2-5.4. The V/K for NADPH, the V/K for 3-APADPH, and the pKi of ATP-ribose also decreased above a pK of 9-10, suggesting interaction of the 2'-phosphate of the nucleotide with a protonated base, perhaps lysine. Since protonation of a residue with a pK of 8 (evident in V/K for DL-glyceraldehyde and V/K for L-gulonate versus pH profiles) appears to be essential for aldehyde reduction, and deprotonation for alcohol oxidation, this residue appears to act as a general acid-base catalyst. An additional anion binding site with a pK of 9.94 facilitates the binding of carboxylic substrates such as D-glucuronate. With NADPH as the coenzyme the primary deuterium isotope effects on V and V/K for NADPH were close to unity and pH independent, suggesting that the hydride transfer step is not rate determining over the experimental pH range. With 3-APADPH as the coenzyme, the maximum velocity, relative to NADPH was three- to four-fold lower. Isotope effects on V, V/K for 3-APADPH, and V/K for D-glucuronate were pH independent and equal to 2.2-2.8, indicating that the chemical step of the reaction is relatively insensitive to pH. These data suggest that substrates bind to both the protonated and the deprotonated forms of the enzyme, though only the protonated enzyme catalyzes aldehyde reduction and the deprotonated enzyme catalyzes alcohol oxidation. On the basis of these results a scheme for the chemical mechanism of aldehyde reductase is postulated.


Increase of the soluble IL-4 receptor (IL-4sR) and positive correlation between IL-4sR and IgE in nasal fluids from school children with allergic rhinitis.
Soluble cytokine receptors (SCR) can either act as inhibitors, by competitively inhibiting cytokines from binding to their membrane-bound receptors, or as enhancers, by serving as cytokine carriers. We have previously found that the levels of the Th2 cytokines interleukin (IL)-4, IL-5, IL-6, and IL-10 were positively correlated to eosinophils and IgE in nasal fluids from 60 children with seasonal allergic rhinitis. In this study, nasal fluids were reexamined to analyze IL-4sR, IL-6sR, IL-1 beta, TNF-alpha, IL-1sR2, TNF-sR1, and TNFsR2 in relation to eosinophils, neutrophils, ECP, and IgE. In allergic patients IL-4sR increased significantly during the pollen season, and weak, but positive correlations with IgE and eosinophils were found (r = 0.45, P < 0.001 and r = 0.4, P < 0.001 respectively). By contrast, none of the other SCR showed increases or correlations with IgE. However, positive correlations between IL1 beta, TNF-alpha, IL-6sR, IL-1sR2, TNF-sR1, TNF-sR2, and either neutrophils or ECP were found. Also, in healthy controls, these cytokines and their receptors were positively correlated to neutrophils or ECP. Thus, increased levels of the soluble IL-4 receptor, as well as IgE, were specifically associated with allergic rhinitis, whereas all other SCR correlated with either inflammatory cells or their products, in both allergic and healthy subjects. These results may suggest that SCR in vivo act as cytokine enhancers, rather than inhibitors.