Contribution of non-volatile and aroma fractions to in-mouth sensory properties of red wines: wine reconstitution strategies and sensory sorting task.
This work explores to what extent the aroma or the non-volatile fractions of red wines are responsible for the overall flavor differences perceived in-mouth. For this purpose, 14 samples (4 commercial and 10 reconstituted wines), were sorted by a panel of 30 trained assessors according to their sensory in-mouth similarities. Reconstituted wines were prepared by adding the same volatile fraction (coming from a red wine) to the non-volatile fraction of 10 different red wines showing large differences in perceived astringency. Sorting was performed under three different conditions: (a) no aroma perception: nose-close condition (NA), (b) retronasal aroma perception only (RA), and (c) allowing retro- and involuntary orthonasal aroma perception (ROA). Similarity estimates were derived from the sorting and submitted to multidimensional scaling (MDS) followed by hierarchical cluster analysis (HCA). Results have clearly shown that, globally, aroma perception is not the major driver of the in-mouth sensory perception of red wine, which is undoubtedly primarily driven by the perception of astringency and by the chemical compounds causing it, particularly protein precipitable proanthocyanidins (PAs). However, aroma perception plays a significant role on the perception of sweetness and bitterness. The impact of aroma seems to be more important whenever astringency, total polyphenols and protein precipitable PAs levels are smaller. Results also indicate that when a red-black fruit odor nuance is clearly perceived in conditions in which orthonasal odor perception is allowed, a strong reduction in astringency takes place. Such red-black fruit odor nuance seems to be the result of a specific aroma release pattern as a consequence of the interaction between aroma compounds and the non-volatile matrix.


A transgenic mouse model for high content, cell cycle phenotype screening in live primary cells.
High content cell-based genetic and small molecule library screens are powerful strategies in drug discovery and investigations of disease mechanisms. We report that primary cells derived from a transgenic mouse model expressing a fluorescence mitosis biosensor provide unambiguous phenotype readouts without the need for transfection or immunocytochemistry. Phenotype profiles of cell cycle disruption and of apoptosis are easily detectable at a single time point selected from time-lapse live fluorescence microscopy. Most importantly, this transgenic mouse model may be crossed with cancer mouse models to derive biosensor-expressing primary cancer cells for use in high content screening strategies targeting discovery of tumor-specific chemotherapeutic compounds.


[Immune complex and vasculitis].
In the early 1900s, von Pirquet first introduced the concept that antigen antibody complexes might cause disease. Since that time, systemic necrotizing vasculitis is believed to be a typical form of immune complex-mediated vasculitis and it may be found in many disorders associated with immunological abnormalities. In 1866, Kusmaul and Maier reported a case with multilesions of necrotizing vasculitis named as periarteritis nodosa which is now nominated as polyarteritis nodosa characterized by the presence of focal inflammatory lesions in small and medium-sized arteries. Besides polyarteritis nodosa, necrotizing vasculitis is frequently found in connective tissue diseases, especially in SLE and RA. Granulomatous vasculitis is a definitely different type of vasculitis from necrotizing vasculitis and known to be found in Wegener's granulomatosis. Whether or not granulomatous vasculitis is an immune complex-mediated disease is unknown, but it can be produced experimentally in several settings. In a experimental model of serum sickness, animals who are hyperimmune responders have widespread granulomatous lesions similar to Wegener's granulomatosis in histology. Intravenous injection of complete Freund's adjuvant in rabbits followed by intravenous administration of human IgM rheumatoid factor is capable of inducing a marked pulmonary granulomatous reaction. The relation of these observations to human Wegener's granulomatosis is uncertain, but the histology and location of the lesions are strikingly similar to human disease. Vasculitis involved in large and medium-sized arteries is seen in two different types of diseases known as temporal arteritis and Takayasu's arteritis. Circulating and tissue-bound immune complexes have been found in both conditions. Most common vasculitis in childhood is Schonlein-Henoch purpura in North America and Kawasaki disease in Japan.(ABSTRACT TRUNCATED AT 250 WORDS)


A real electro-magnetic placebo (REMP) device for sham transcranial magnetic stimulation (TMS).
There is growing interest in neuropsychiatry for repetitive transcranial magnetic stimulation (rTMS) as a neuromodulatory treatment. However, there are limitations in interpreting rTMS effects as a real consequence of physiological brain changes or as placebo-mediated unspecific effects, which may be particularly strong in psychiatric patients. This is due to the fact that existing sham rTMS procedures are less than optimal. A new placebo tool is introduced here, called real electro-magnetic placebo (REMP) device, which can simulate the scalp sensation induced by the real TMS, while leaving both the visual impact and acoustic sensation of real TMS unaltered. Physical, neurophysiological and behavioural variables of monophasic and biphasic single-pulse TMS and biphasic 1Hz and 20Hz rTMS procedures (at different intensities) were tested in subjects who were expert or naïve of TMS. Results of the real TMS were compared with those induced by the REMP device and with two other currently used sham procedures, namely the commercially available Magstim sham coil and tilting the real coil by 90 degrees . The REMP device, besides producing scalp sensations similar to the real TMS, attenuated the TMS-induced electric field (as measured by a dipole probe) to a biologically inactive level. Behaviourally, neither expert nor naïve TMS subjects identified the "coil at 90 degrees " or the "Magstim sham coil" as a real TMS intervention, whilst naïve subjects were significantly more likely to identify the REMP-attenuated TMS as real. The "goodness of sham" of the REMP device is demonstrated by physical, neurophysiological, and behavioural results. Such placebo TMS is superior to the available sham procedures when applied on subjects naïve to TMS, as in case of patients undergoing a clinical rTMS trial.


The impact of neonatal morbidities on child growth and developmental outcomes in very low birth weight infants: a nationwide cohort study.
Growth in preterm infants has long-term implications for neurodevelopmental outcomes. We aimed to estimate the nationwide growth outcomes from birth to 5 years in infants born under 1500 g and to analyze the effects of major morbidities in preterm infants on growth. In total, 2961 children born in 2013 with a birth weight under 1500 g who underwent an infant health checkup between 2013 and 2018 according to the National Health Insurance Service database were included. Checkups were conducted at 4-6, 9-12, 18-24, 30-36, 42-48, and 54-60 months of age. Information was obtained from the International Classification of Diseases-10 codes or a questionnaire administered during the check-up. At 60 months of age, the mean percentiles of weight, height, and head circumference fell within only the 30-40th percentile of normal growth values. About 30% of infants had growth parameters below the 10th percentile and showed worse neurodevelopmental outcomes. Using multiple logistic regression, infants with bronchopulmonary dysplasia showed a significantly higher incidence of growth restriction in all three categories of weight (odds ratio [OR] 1.50), height (OR 1.33), and head circumference (OR 1.36) at 60 months. Sepsis was associated with growth restriction in weight (OR 1.43) and head circumference (OR 1.33). Periventricular leukomalacia infants had relatively small head circumferences (OR 1.91) and poor developmental screening results (OR 2.89).Conclusion: Catch-up growth remains a major issue in infants born under 1500 g, especially those with some morbidities from preterm birth. Regular checkups to monitor and early intervention to achieve normal growth are essential. What is Known: • Growth in preterm infants has long-term implications for neurodevelopmental and cardiometabolic outcomes. • Data are lacking on the time-serial effects of many preterm morbidities simultaneously on long-term growth outcomes. What is New: • All growth parameters of VLBW infants, including weight, height, and head circumference, fell within the 30-40th percentile of normal growth for infants at 60 months of age, indicating that catch-up growth for VLBW infants remains an issue. • VLBW infants with major preterm morbidities, including BPD, PVL, and sepsis, showed difficulties in achieving normal catch-up growth and neurodevelopment at 60 months of age.


Magnetic resonance imaging of bladder pheochromocytomas: a review.
Bladder pheochromocytomas (PCCs) are rare tumors that account for 0.06% of all bladder tumors and makeup 1% of all PCCs. Most PCCs are functional, and they secrete catecholamines that lead to clinical symptoms such as paroxysmal hypertension, headaches, palpitations, and sweating. However, some are nonfunctional and asymptomatic and are hence difficult to diagnose. Cystoscopy and biopsy should not be performed when bladder PCCs are suspected. They may provoke a hypertensive crisis if preventative antiadrenergic blockers are not administered prior to the procedure. The diagnostic workup begins with obtaining blood or urine catecholamine and catecholamine metabolite values to make a presumptive diagnosis of bladder PCC. Computed tomography (C.T.) and magnetic resonance imaging (MRI) are then used to localize and stage the tumor for surgical resection. MRI, due to its superior soft tissue resolution and the ability to use multiparametric MRI (mpMRI) to differentiate between layers of the bladder wall and from other bladder masses, is the optimal imaging modality to detect extra-adrenal bladder PCCs and determine locoregional staging. Once antiadrenergic medications are given, the tumor is resected, and the diagnosis is confirmed histologically. However, the differential diagnosis of bladder PCC often gets overlooked, leading to surgical resection in the absence of antiadrenergic medications, increasing the chances of a fatal hypertensive crisis. This makes MRI an essential diagnostic tool for staging bladder PCCs before surgery. This review discusses the indications for MRI in bladder PCCs and describes findings from these tumors on various MRI sequences and when to use them. We also discuss how MRI can differentiate bladder PCCs from other bladder neoplasms.


Possible targeting of G protein coupled receptors to manipulate inflammation in vivo using synthetic and natural ligands.
Cyclic AMP elevating Gs protein coupled receptors were considered for a long time to be immunosuppressive. One of these receptors, adenosine A(2A) receptor, was implicated in a physiological mechanism that down regulates inflammation and protects tissues from excessive immune mediated damage. Targeting of these receptors by selective agonists may lead to better protocols of anti-inflammatory treatments. At the same time inhibiting the Gs protein coupled mediated signalling with antagonists could be explored in studies of approaches to enhance inflammation and tissue damage. Enhancement of targeted tissue damage is highly desirable when it is cancerous tissue, while enhancement of inflammatory events might be desirable in the development of new vaccine adjuvants.


Evaluation of animal models for the hair-inducing capacity of cultured human dermal papilla cells.
The dermal papilla (DP) interacts with epithelial cells for folliculogenesis. For translational research on cell therapies for hair regrowth with cultured human DP cells (hDPCs), a model to evaluate the capacity of hDPCs to induce hair formation is inevitable. Chamber models were constructed by transplanting 4 different combinations of mouse or human epithelial and mesenchymal cells into a silicone chamber implanted onto the back of nude mice. In parallel, 3 types of sandwich constructs were created by inserting hDPCs or human DP tissue between the epidermis and dermis of isolated rat footpad skin or human facial skin, and subcutaneously transplanting the constructs into the back of nude mice. Four to six weeks later, skin sections of each model were histologically examined. Folliculoneogenesis was detected in both chamber and sandwich models, although the induction rate and maturity of the hair follicles varied among cell combination subgroups in each model. The difference in hair induction rate was not statistically significant between 2 representative chamber and sandwich subgroups using cultured hDPCs. The sandwich model, however, required fewer hDPCs, did not require human keratinocytes, and exhibited a higher rate of successful sample collection. Although there is no significant difference in hair induction rate, the sandwich model using cultured hDPCs and the rat sole skin is more feasible than the chamber model using human cultured keratinocytes and hDPCs as a tool to evaluate the hair-inducing capacity of cultured hDPCs.


Theoretical study of the mechanism of protein arginine deiminase 4 (PAD4) inhibition by F-amidine.
Protein arginine deiminase 4 (PAD4) catalyzes the hydrolysis of a peptidylarginine residue to form a citrulline residue and ammonia during posttranslational modification. This process plays a pivotal role in rheumatoid arthritis (RA) and gene regulation. F-amidine belongs to a series of haloacetamidine compounds that are the most potent PAD4 inhibitors described to date. F-amidine acts as a mechanism-based inhibitor of PAD4, inactivating PAD4 by the covalent modification of the active site Cys645. In this manuscript, the fundamental mechanism of PAD4 inhibition by F-amidine is investigated using a QM/MM approach. Our simulations show that in the PAD4-F-amidine reactant complex, the active site Cys645 exists as a thiolate and His471 is protonated. This is consistent with the reverse protonation mechanism wherein the active site nucleophile, Cys645, in PAD4 exists as a thiolate in the active form of the enzyme. Inhibition of PAD4 by F-amidine is initiated by the nucleophilic addition of Sγ to the Cζ of F-amidine, leading to the formation of a tetrahedral intermediate. His471 serves as a proton donor, helping F to leave the fluoroacetamidine moiety of F-amidine; meanwhile, Sγ forms a three-membered ring with Cζ and Cη of F-amidine. Subsequently, the three-membered sulfonium ring collapses and rearranges to the final thioether product. His471 acts as a proton donor in the transition state and facilitates the inhibition reaction of PAD4.


Differential activation of pregnane X receptor by carnosic acid, carnosol, ursolic acid, and rosmarinic acid.
Pregnane X receptor (PXR) regulates the expression of many genes, including those involved in drug metabolism and transport, and has been linked to various diseases, including inflammatory bowel disease. In the present study, we determined whether carnosic acid and other chemicals in rosemary extract (carnosol, ursolic acid, and rosmarinic acid) are PXR activators. As assessed in dual-luciferase reporter gene assays, carnosic acid, carnosol, and ursolic acid, but not rosmarinic acid, activated human PXR (hPXR) and mouse PXR (mPXR), whereas carnosol and ursolic acid, but not carnosic acid or rosmarinic acid, activated rat PXR (rPXR). Dose-response experiments indicated that carnosic acid, carnosol, and ursolic acid activated hPXR with EC50 values of 0.79, 2.22, and 10.77μM, respectively. Carnosic acid, carnosol, and ursolic acid, but not rosmarinic acid, transactivated the ligand-binding domain of hPXR and recruited steroid receptor coactivator-1 (SRC-1), SRC-2, and SRC-3 to the ligand-binding domain of hPXR. Carnosic acid, carnosol, and ursolic acid, but not rosmarinic acid, increased hPXR target gene expression, as shown by an increase in CYP3A4, UGT1A3, and ABCB1 mRNA expression in LS180 human colon adenocarcinoma cells. Rosmarinic acid did not attenuate the extent of hPXR activation by rifampicin, suggesting it is not an antagonist of hPXR. Overall, carnosic acid, carnosol, and ursolic acid, but not rosmarinic acid, are hPXR agonists, and carnosic acid shows species-dependent activation of hPXR and mPXR, but not rPXR. The findings provide new mechanistic insight on the effects of carnosic acid, carnosol, and ursolic acid on PXR-mediated biological effects.


Co-delivery of three synergistic chemotherapeutics in a core-shell nanoscale coordination polymer for the treatment of pancreatic cancer.
The combination chemotherapy regimen FOLFIRINOX comprising folinic acid, 5-fluorouracil, irinotecan, and oxaliplatin is the first-line treatment for patients with advanced pancreatic cancer, but its use remains prohibitive for the majority of patients due to severe side effects. Here, we report a core-shell nanoscale coordination polymer (NCP) nanoparticle co-delivering a potent and synergistic combination of oxaliplatin, gemcitabine, and SN38 (OGS), for the treatment of pancreatic cancer in mouse models. OGS contains key synergistic components of FOLFIRINOX in a controllable drug ratio., It exhibited particle stability in blood circulation and enhanced deposition of the drugs in acidic tumor environments. In vitro, OGS showed superior cytotoxicity over free drug combinations and robust cytotoxic synergism among its three components. In vivo, OGS improved drug circulation, increased tumor deposition, and exhibited superior antitumor efficacy over the free drug combination in both subcutaneous and orthotopic pancreatic tumor models. OGS treatment achieved 75-91% tumor growth inhibition and prolonged mouse survival by 1.6- to 2.8-folds while minimizing systemic toxicities such as neutropenia, hepatotoxicity, and renal toxicity. This work uncovers a novel and clinically relevant nanomedicine strategy to co-deliver synergistic combination chemotherapies for difficult-to-treat cancers.


Prognostic biomarkers for sepsis mortality based on the literature and LC-MS-based metabolomics of sepsis patients.
The management of sepsis, a potentially lethal overreaction to infection, is limited by the lack of prognostic tools to guide its treatment. Our aim is to identify a novel metabolic biomarker panel for predicting sepsis mortality based on a literature review and liquid chromatography-mass spectrometry (LC-MS)-based metabolomics. In the literature, we found metabolomics biomarkers reported to predict sepsis mortality. We determined the classifications, reported frequency, and KEGG pathway enrichment of these markers. Using serum samples from 20 sepsis survivors and 20 non-survivors within 28 days after admission to the intensive care unit (ICU), we performed LC-MS-based metabolomics. Based on the literature review and metabolomics, a prognostic biomarker panel for sepsis was identified and its area under the curve (AUC) values was assessed. Kynurenate, caffeine, and lysoPC 22:4 were selected as a prognostic biomarker panel for sepsis. The panel had an area under the curve (AUC) of 0.885 (95% CI, 0.694-1) evaluated by linear support vector machine (SVM) and 0.849 (0.699-1) by random forest (RF), which was higher than that of the Sequential Organ Failure Assessment (SOFA). A combination of kynurenate, caffeine, and lysoPC 22:4 and SOFA provided the best discriminating performance, with AUCs of 0.961 (0.878-1) for SVM and 0.916 (0.774-1) for RF. The prognostic biomarker panel consisting of kynurenate, caffeine, and lysoPC 22:4 may aid in the identification of sepsis patients at a high risk of death, leading to personalized therapy in clinical practice that will improve sepsis survival.


Plasticity varies with boldness in a weakly-electric fish.
The expression of animal personality is indicated by patterns of consistency in individual behaviour. Often, the differences exhibited between individuals are consistent across situations. However, between some situations, this can be biased by variable levels of individual plasticity. The interaction between individual plasticity and animal personality can be illustrated by examining situation-sensitive personality traits such as boldness (i.e. risk-taking and exploration tendency). For the weakly electric fish Gnathonemus petersii, light condition is a major factor influencing behaviour. Adapted to navigate in low-light conditions, this species chooses to be more active in dark environments where risk from visual predators is lower. However, G. petersii also exhibit individual differences in their degree of behavioural change from light to dark. The present study, therefore, aims to examine if an increase of motivation to explore in the safety of the dark, not only affects mean levels of boldness, but also the variation between individuals, as a result of differences in individual plasticity. Boldness was consistent between a novel-object and a novel-environment situation in bright light. However, no consistency in boldness was noted between a bright (risky) and a dark (safe) novel environment. Furthermore, there was a negative association between boldness and the degree of change across novel environments, with shier individuals exhibiting greater behavioural plasticity. This study highlights that individual plasticity can vary with personality. In addition, the effect of light suggests that variation in boldness is situation specific. Finally, there appears to be a trade-off between personality and individual plasticity with shy but plastic individuals minimizing costs when perceiving risk and bold but stable individuals consistently maximizing rewards, which can be maladaptive.


Effects of nitric oxide on bovine polymorphonuclear functions.
The effects of nitric oxide (NO) on the functionality of polymorphonuclear neutrophils (PMNs) in bovine milk or blood were investigated. In 2 experiments, mastitis was induced by infusing both hind quarters with saline containing Escherichia coli endotoxins. In addition, the left hind quarter was infused with aminoguanidine, an inhibitor of the inducible form of NO synthase (iNOS). At various times after infusion, somatic cells were isolated from milk samples, and superoxide (O2-) production induced by phorbol myristate acetate was evaluated. In both experiments, the addition of aminoguanidine had no inhibitory effect on the number of milk somatic cells or on their O2- production. The effect of NO and iNOS inhibitors on the functionality of bovine PMNs isolated from blood was investigated in vitro. The neutrophils did not produce NO. A neutrophil:monocyte co-culture system was used to study the effect of NO derived from monocytes on O2- production by bovine neutrophils. Neither NO derived from activated monocytes nor the iNOS inhibitors aminoguanidine and L-N6-(1-iminoethyl)lysine had an effect on the ability of bovine neutrophils to release O2-. Moreover, aminoguanidine did not affect the ability of bovine neutrophils to phagocytose bacteria. These results suggest that inhibition of NO release during inflammation does not interfere with the migration of immune cells to the site of infection or the ability of these cells to destroy pathogens. Thus, NO does not appear to play a major role in the control of the functions of bovine neutrophils.


A visible-light harvesting system for CO2 reduction using a Ru(II) -Re(I) photocatalyst adsorbed in mesoporous organosilica.
A photocatalytic system for CO2 reduction exhibiting visible-light harvesting was developed by preparing a hybrid consisting of a supramolecular metal complex as photocatalyst and periodic mesoporous organosilica (PMO) as light harvester. A Ru(II) Re(I) binuclear complex (RuRe) with methylphosphonic acid anchor groups was adsorbed on acridone or methylacridone embedded in the walls of PMO mesochannels to yield the hybrid structure. The embedded organic groups absorbed visible light, and the excitation energy was funneled to the Ru units. The energy accumulation was followed by electron transfer and catalytic reduction of CO2 to CO on the Re unit. The light harvesting of these hybrids enhanced the photocatalytic CO evolution rate by a factor of up to ten compared with that of RuRe adsorbed on mesoporous silica without a light harvester.


The activity of PS-15 in combination with sulfamethoxazole.
PS-15 is a novel biguanide folate antagonist that is metabolized in vivo to WR99210, a metabolite that is extremely active in vitro against multi-drug resistant strains of Plasmodium falciparum. When PS-15 was administered in combination with sulfamethoxazole to healthy Saimiri sciureus monkeys, the serum antimalarial activity was considerably greater than that observed in monkeys that received PS-15 alone. Further studies should be carried out to determine the value of PS-15/sulfonamide combinations in the treatment of human malaria infections and in preventing the emergence of drug-resistant parasites.


Molecular mechanisms of cholesteryl ester transfer protein deficiency in Japanese.
Plasma cholesteryl ester transfer protein (CETP) facilitates the transfer of cholesteryl ester (CE) from high density lipoprotein (HDL) to apolipoprotein B-containing lipoproteins. Since CETP regulates the plasma levels of HDL cholesterol and the size of HDL particles, CETP is considered to be a key protein in reverse cholesterol transport (RCT), a protective system against atherosclerosis. The importance of plasma CETP in lipoprotein metabolism was demonstrated by the discovery of CETP-deficient subjects with marked hyperalphalipoproteinemia (HALP). Genetic CETP deficiency is the most important and common cause of HALP in the Japanese. Ten mutations of the CETP gene have been demonstrated as causes of HALP, including two common mutations: an intron 14 splicing defect (Int14 + 1 G --> A) and an exon 15 missense mutation (D442G). The subjects with CETP deficiency show a variety of abnormalities in the concentration, composition, and function of both HDL and low density lipoprotein (LDL). CETP deficiency is considered a physiological state of impaired RCT, which may possibly lead to the development of atherosclerosis despite high HDL cholesterol levels. However, the pathophysiological significance of CETP in terms of atherosclerosis has been controversial. Epidemiological studies in Japanese-Americans living in Hawaii and Japanese in the Omagari area, where HALP subjects with an intron 14 splicing defect of the CETP gene are markedly frequent, have shown a relatively increased incidence of coronary atherosclerosis in CETP deficiency. On the other hand, the TaqIB polymorphism-B2 allele with low CETP mass and increased HDL cholesterol has been related to a decreased risk for coronary heart disease (CHD) in many studies, including the Framingham Offspring Study. The current review focused on the characterization of the Japanese subjects with CETP deficiency, including our recent findings.


Role of leukocytes and leukocyte adhesion molecules in renal ischemic-reperfusion injury.
Renal ischemic-reperfusion injury (IRI) occurs frequently in transplanted as well as native kidneys. Effective treatment for this process is still elusive. Leukocytes and their products may be important in the pathogenesis of renal IRI, however their role is still controversial. Recently, adhesion receptors on leukocytes and their corresponding ligands have been identified. In the heart, considerable evidence supports the role of CD11/CD18, ICAM-1, and the selectin receptors in IRI. However, based on experimental studies in animal models, even though renal IRI appears to be ICAM-1 mediated, the role of the CD11/CD18 pathway appears to be minimal. In addition, the available evidence does not support the concept that L-selectin has significant involvement in renal IRI. In this review, the data and controversies regarding the role of leukocytes and leukocyte adhesion molecules in renal IRI are discussed.


Structure and function of adenylate kinase isozymes in normal humans and muscular dystrophy patients.
Two isozymes of adenylate kinase from human Duchenne muscular dystrophy serum, one of which was an aberrant form specific to DMD patients, were separated by Blue Sepharose CL-6B affinity chromatography. The separated aberrant form possessed a molecular weight of 98,000 +/- 1,500, whereas the normal serum isozyme had a weight of 87,000 +/- 1,600, as determined by SDS-polyacrylamide gel electrophoresis, gel filtration, and sedimentation equilibrium. The sedimentation coefficients were 5.8 S and 5.6 S for the aberrant form and the normal form, respectively. Both serum isozymes are tetramers. The subunit size of the aberrant isozyme (Mr = 24,700) was very similar to that of the normal human liver isozyme, and the subunit size of the normal isozyme (Mr = 21,700) was very similar to that of the normal human muscle enzyme. The amino acid composition of the normal serum isozyme was similar to that of the muscle-type enzyme, and that of the aberrant isozyme was similar to that of the liver enzyme, with some exceptions in both cases.


Nicotine intake and problem solving strategies are modified during a cognitively demanding water maze task in rats.
Nicotine is the major addictive component in tobacco, and despite well-established adverse health effects of tobacco addiction, some smokers have difficulty quitting. The acute cognitive enhancement and/or the amelioration of the cognitive disruption during withdrawal that some smokers experience after smoking are among important factors that hinder quit attempts. The animal model presented in the current study is comparable to the human smoking condition although nicotine intake routes are different. Rats were exposed to a free choice of oral nicotine starting at adolescence, and given a water maze (WM) task as adults. This design allowed us to see if rats alter their nicotine intake during the WM task and if nicotine preference and intake modify abilities and strategies rats use for problem solving. Male and female rats were exposed to a free choice of oral nicotine/water for 24weeks, starting at five weeks of age. After this period, they were selected based on their nicotine intake and, together with control animals that received only water, were subjected to a place-learning task in the WM. Free-choice nicotine exposure continued during WM testing. Following acquisition, the probe trial presented the rats with a choice between using two different strategies for problem solving. Nicotine supported acquisition and rats increased their nicotine intake during WM testing; this effect was more pronounced in male rats with minimum nicotine preference and intake. Furthermore, nicotine modified the "female type" strategy in solving the place-learning task and nicotine treated female rats, unlike control females, behaved like males. The increase in nicotine intake during mental engagement, and the sexually dimorphic effect of nicotine on problem solving strategies that we have observed in rats, may suggest that implementing sex-specific smoking cessation approaches, especially under stressful and cognitively demanding conditions, may be useful in helping smokers quit.


Coherent Plasmon and Phonon-Plasmon Resonances in Carbon Nanotubes.
Carbon nanotubes provide a rare access point into the plasmon physics of one-dimensional electronic systems. By assembling purified nanotubes into uniformly sized arrays, we show that they support coherent plasmon resonances, that these plasmons couple to nanotube and substrate phonons, and that the resulting phonon-plasmon resonances have quality factors as high as 10. Because nanotube plasmons intensely strengthen electromagnetic fields and light-matter interactions, they provide a compelling platform for surface-enhanced spectroscopy and tunable optical devices at deep-subwavelength scales.


[N-isopropyl-p-[123I]iodoamphetamine early dynamic image with ring-type emission CT in cerebrovascular disease].
N-isopropyl-p-[123I]iodoamphetamine (IMP) SPECT was done in 14 patients with cerebrovascular disease. Immediately after injection of 3 mCi IMP, sequential 2 minutes SPECT images (dynamic images) were obtained for 15 minutes using a circular detector array emission tomographic system (Shimadzu SET-030W). Then standard SPECT images were obtained at 30 minutes (early images) and 5 hours (delayed images) using a rotating scintillation camera ECT system (GCA-601E, Toshiba). Two patients out of 14 studies showed discrepancy of the findings between dynamic images and early images; dynamic images revealed low uptake area though early images showed no defect. Possible mechanism of this phenomenon might be due to so called redistribution phenomenon occurred during obtaining early images. In conclusion, early dynamic images is useful in cases with slightly decreased cerebral blood flow.


Evaluation of the DipStreak, a new device with an original streaking mechanism for detection, counting, and presumptive identification of urinary tract pathogens.
DipStreak is a new urine culture device with two types of agar attached back-to-back on a plastic paddle. It combines dip-slide technology and an original streaking inoculation mechanism, allowing for bacterial counting and colony isolation. The performance of the DipStreak device with two different medium formulations, CHROMagar and MacConkey media in study A and UriSelect 3 and MacConkey media in study B, was evaluated and compared to that of the reference streak method by using plates of cystine-lactose-electrolyte-deficient (CLED) agar, tryptic soy agar with 5% sheep blood, and UriSelect 3 medium. In study A, 2,000 urine specimens were processed and 511 cultures were found positive. The DipStreak device and the UriSelect 3 and CLED medium plates gave the same detection rate, 99.7%. For the direct identification of Escherichia coli, Proteus mirabilis, and Enterococcus sp. isolates, the DipStreak device and the UriSelect 3 medium plate showed overall sensitivities of 97 and 93.4%, respectively. In study B, 3,000 urine specimens were processed and 714 cultures were found positive. The DipStreak device and the UriSelect 3 and CLED medium plates gave detection rates of 99.4, 99.9, and 99.2%, respectively. For the direct identification of E. coli, P. mirabilis, and Enterococcus sp. isolates, the DipStreak device and the UriSelect 3 medium plate showed overall sensitivities of 88 and 94.4%, respectively. In conclusion, the DipStreak device with both medium formulations represents an attractive and excellent screening method for the reliable detection, counting, and presumptive identification of urinary tract pathogens. It enables bedside urine inoculation and provides a valid means of transporting the sample back to the laboratory, decreasing drastically the rate of false-positive results due to bacterial overgrowth and reducing associated costs.


Infectivity enhancement of different HIV-1-based lentiviral pseudotypes in presence of the cationic amphipathic peptide LAH4-L1.
Lentiviral vectors (LVs) are promising delivery systems for gene therapy. To enhance the efficiency of target cell transduction by LVs, protocols often include the addition of culture additives. In this study, the cationic amphipathic peptide LAH4-L1 (KKALLAHALHLLALLALHLAHALKKA), a DNA transfection agent, was evaluated for its capacity to improve LV infectivity in cell lines and primary cells. Results show that LAH4-L1 enhances infectivity of all LV pseudotypes tested, particularly GALVTR-LVs. More importantly, LAH4-L1 promotes the transduction of CD34+ hematopoietic stem cells with GALVTR-LVs as efficiently as Retronectin, a culture additive used in ex vivo clinical protocols involving LVs. The action of LAH4-L1 relies both on the GALVTR-LV adhesion and post-adhesion steps. LAH4-L1 represents a new and attractive transduction enhancer for hematopoietic gene therapy protocols.


Enhanced guided bone regeneration with a resorbable chamber containing demineralized bone matrix.
The effectiveness of a nonporous poly-DL-lactide tubular chamber in guiding bone regeneration through a long bone defect had already been assessed in an experimental model using the rabbit radius. The injection of bone marrow stem cells into the chamber had proven to enhance bone regeneration. The present study reports on the development of the above research project in a subsequent stage. Demineralized bone matrix (DBM) obtained by milling New Zealand rabbit femoral and tibial diaphyses was placed into a tubular chamber. A 10-mm defect was bilaterally created in the radii of 10 rabbits. On the left side (chamber side) the defect was treated by means of a poly-DL-lactide chamber filled with DBM, whereas DBM alone was used on the right side (control). Controls were performed at 3 and 6 months by radiographs and histomorphometry and demonstrated better bone growth on the chamber side versus the control side. A comparison with the results previously obtained by stem cell injection into the chamber revealed significant acceleration of bone regrowth in the first 3 months because of the addition of DBM to the chamber. However, no significant difference was found between the two sides after 6 months. These results have confirmed the effectiveness of the chamber as a container for the factors promoting bone regeneration, probably because the osteogenetic activity is maintained in situ.


Hatchet-shaped tensor fascia lata musculocutaneous flap for the coverage of trochanteric pressure sores: a new modification.
The tensor fascia lata flap is one of the appropriate choices for the coverage of trochanteric pressure sores. The authors designed a new, hatched-shaped tensor fascia lata musculocutaneous flap with distal Z-plasty closure and applied it to four trochanteric defects in 4 patients. Satisfactory results were obtained in all patients. The hatchet-shaped tensor fascia lata musculocutaneous flap is very safe, reliable, and practical. Designing the flap in a hatchet shape allows one to use the proximal and well-vascularized portion of the flap in the trochanteric pressure sore area. Another important advantage is the possibility of reuse resulting from recurrence. Prevention of a "dog-ear" deformity at the recipient site provides a smooth contour on the lateral aspect of the thigh. Another advantage is the tension-free Z-plasty closure of the donor site without need of grafting.


Acute immuno-allergic interstitial nephritis caused by fluindione.
Fluindione is a vitamin K antagonist that is commonly prescribed for the treatment of cardiovascular disease and venous thromboembolism in France. Bleeding is the most common side effect of fluindione, whereas hypersensitivity reactions are rare. We describe here a patient with acute immuno-allergic interstitial nephritis caused by fluindione. Initial symptoms included fever, eosinophilia, low albuminuria, microscopic hematuria, eosinophiluria and acute renal failure. Kidney biopsy showed severe interstitial nephritis with interstitial edema, inflammatory infiltrates and tubulorrhexis. Fluindione withdrawal and corticosteroid treatment resulted in rapid recovery of renal function. A review of the literature revealed a very low incidence of fluindione-induced interstitial nephritis, with variable renal and extra-renal signs. Early recognition of this rare complication may prevent the development of severe chronic renal injury.


Gene activation by the Escherichia coli positive regulator, OmpR. Phosphorylation-independent mechanism of activation by an OmpR mutant.
In Escherichia coli, expression of the major outer membrane proteins, OmpC and OmpF, is regulated through the functions of OmpR and EnvZ at the transcriptional level in response to the medium osmolarity. OmpR is the crucial activator that helps RNA polymerase to efficiently trigger ompC and ompF transcription. This OmpR function is modulated by phosphorylation mediated by the cognate sensory kinase, EnvZ. Phosphorylation at the N-terminal domain of OmpR results in substantial enhancement of the DNA-binding ability of the C-terminal domain, thereby allowing the activation of ompC and ompF transcription by OmpR. Here we isolated an OmpR mutant which lacks the N-terminal half, but can enhance transcription in vivo. This novel type of OmpR mutant was revealed to have a single amino acid replacement of Gly227 to Cys. The newly-introduced-Cys residue allows OmpR molecules to form a stable dimer in vitro without the help of the N-terminal half. This altered C-terminal half is able to bind efficiently and specifically to the cognate DNA in vitro. It can function as an activator for ompC transcription in vitro in a phosphorylation-independent manner. These results suggest that the putative activator domain of OmpR, together with the DNA-binding domain, is most likely located in the C-terminal half. They also suggested that the phosphorylation of OmpR may not be essential for gene activation per se.


Oestrogen receptor low positive breast cancer: associations with prognosis.
In this study of oestrogen receptor (ER) Low Positive breast cancers (BC) in three large cohorts of BC patients, we assess associations between levels of ER expression and tumour characteristics and prognosis. Cases were stratified into patients unlikely to have received adjuvant therapy according to treatment guidelines at time of diagnosis (before 1995), and those who could have received adjuvant therapy (diagnosed in 1995 or later). ER status was divided into < 1%; ≥ 1 < 10%; ≥ 10%. Results were correlated with time of diagnosis, histopathological grade, proliferation status, and molecular subtypes, using Pearson's Chi-square test. For prognosis, hazard ratios and cumulative incidence of death from BC were used. Of the 1955 tumours, 65 (3.3%) were ER Low Positive (ER ≥ 1 < 10%). Overall, the highest proportion of ER Low Positive tumours was observed among Luminal B (HER2 +) subtype (9.4%) and grade 3 tumours (4.3%). The risk of death from BC was lower in ER Low Positive and ER ≥ 10% compared to ER-negative cases. Compared to patients diagnosed before 1995, women diagnosed in 1995 or later showed a higher proportion of ER Low Positive BCs, and their tumours were of smaller size, lower grade, and lower proliferative status. There was no significant difference in prognosis compared to those with ER ≥ 10% tumours. Women with ER Low Positive tumours diagnosed in a time period when adjuvant therapy was available had tumours of smaller size, lower grade, and lower proliferative status, and similar prognosis to those with ER ≥ 10% compared to women diagnosed earlier.


[Interpretation of the concept of 'medicinal product' in relation to herb- and cannabinoid-based products].
On 10th of July 2014 the European Court of Justice made in his decisions in relation to the cases D. (C-358/13) and G. (C-181/14) an interpretation, that the concept of 'medicinal product' according to the law of the European Union does not include the materials, which are - as not covering substances, such as those at issue in the main proceedings, which produce effects that merely modify physiological functions but which are not such as to have any beneficial effects, either immediately or in the long term, on human health, are consumed solely to induce a state of intoxication and are, as such, harmful to human health. The Court made his interpretation after the request for preliminary ruling from the Bundesgerichtshof (the High Court of Justice in Germany). The Court had to decide in two criminal procedures, whether for the retail of mixtures including syntetic canabinoids, such as complements of marihuana, due the fact that they are "unsafe medicinal products", a criminal proceeding can be initiated or not. The Ordinary Courts had two persons (D. and G.) for selling the unsafe medicinal products sentenced to one year and nine months imprisonment, and suspension (D.), and sentenced (G.) to four years and six months imprisonment and fined with a charge of two hundred thousand Euro. The retail of herb mixtures containing, inter alia, synthetic cannabinoids, did not fall under the German law on narcotic drugs at the material time, resulting that the German Authorities could not initiate a criminal procedure.


[The effects of Xiaotan Huayu Liqiao formula on mesenteric artery function in rats exposed to chronic intermittent hypoxia].
To investigate the effects of Xiaotan Huayu Liqiao formula (the Chinese Medicine) on mesenteric artery function in rats exposed to chronic intermittent hypoxia (CIH), and to explore the related mechanism. Forty-eight male SD rats were randomly divided into four groups as Normoxia, CIH, Formula+CIH and formula group. Rats were exposed to normoxia in the Normoxia and Formula group, or intermittent hypoxia in CIH or Formula+CIH group. Xiaotan Huayu Liqiao formula was given at 24g/kg by intragastric administration before intermittent hypoxia exposure. The pathological changes of mesenteric artery were determined by HE staining, and the relaxation of mesenteric artery (induced by acetylcholine(ACh) and L-arginine(L-Arg)) was recorded by microvascular ring technique. Serums of all rats were collected (0 d and 21 d) and the content of NO was detected by ELISA. The levels of endothelial nitric oxide synthase (eNOS) and p-eNOS were measured by Western blot. Compared with Normoxia group, the mesenteric arterial endothelial injury and media thickening were observed and the relaxation of mesenteric artery was significantly reduced in rats exposed to CIH. The level of NO in serum and the ratio of p-eNOS/eNOS were also decreased in the CIH group. Xiaotan Huayu Liqiao formula administration improved the pathologic changes and dilatation function of mesenteric artery, increased the levels of NO and p-eNOS. Compared with Normoxia group,all the results were not observed significant difference in Formula group. Xiaotan Huayu Liqiao formula increased the bioavailability of NO, and ameliorated the CIH induced mesenteric artery function injury.


Effects of dietary copper (II) sulfate and copper proteinate on performance and blood indexes of copper status in growing pigs.
160 crossbred (Duroc x Landrace x Yorkshire) gilts averaged 21.25 kg body weight were used to study the effects of dietary copper (II) sulfate (CuSO4) and copper proteinate (Cu-Pr) on growth performance, plasma Cu concentration, ceruloplasmin activity, and erythrocyte Cu/Zn-superoxide dismutase (SOD) activity. All pigs were allotted to four treatments and fed with basal diets supplemented with 0 (control), 250 mg /kg Cu as CuSO4, and 50 and 100 mg/kg Cu as Cu-Pr. Growth performance was determined based on two growth phase (phase 1: days 0 to 15, phase 2: days 15 to 30). After 30 days of the treatment, 16 pig blood samples (four per treatment) were collected for indexes of copper status determination. The experimental results showed that compared with control, pigs fed with 250 mg Cu/kg as CuSO4 and 100 mg Cu/kg as Cu-Pr had higher average daily gain and average daily feed intake in the whole growth phase (d 0 to 30). In addition, 250 mg Cu/kg as CuSO4 and 100 mg/kg Cu as Cu-Pr enhanced plasma ceruloplasmin activity (P < 0.05), and 100 mg/kg Cu as Cu-Pr increased erythrocyte Cu/Zn-SOD activity (P < 0.01) compared with the control. There was no obvious treatment response on plasma Cu concentration in the present study.


A case of pulmonary embolism caused by paroxysmal nocturnal hemoglobinuria.
A patient with pulmonary embolism caused by paroxysmal nocturnal hemoglobinuria (PNH) is described. PNH sometimes causes venous thrombosis in the extremities, hepatic vein, or cerebral vein, but pulmonary embolism rarely occurs.


Development and differentiation of the reproductive system of Tropidurus catalanensis (Squamata: Tropiduridae).
Development and differentiation of the reproductive system in lizards begin in the embryonic period, although the stage and time of their occurrence vary according to populations and species. In this study, the events of the development and differentiation of the reproductive system of males and females of Tropidurus catalanensis were characterized during the embryonic, neonatal, and juvenile periods. Embryos at Stages 27, 34, 37, 40, and 41, neonates and juveniles, from Corrientes, Argentina, were analyzed. At Stage 27, the genital ridge was not observed but primordial germ cells were recorded in the yolk sac as well as the mesenteric mesenchyme, indicating the beginning of germ cell migration. Gonadal differentiation commenced at Stage 34. In males from Stage 37, the testes possessed seminiferous cords with Sertoli cells and spermatogonia, while in hatchlings seminiferous tubules and interstitial tissue with mature Leydig cells were present. Spermatogenesis was observed in a specimen of 51.9 mm snout-vent length, corresponding to the minimum reproductive size. In females, from Stage 37 until hatching, the ovaries had a cavernous medulla and a cortex with somatic cells and abundant oogonia. The onset of meiosis and folliculogenesis occurred in the juvenile period.


Characterizing stability properties of supported bilayer membranes on nanoglassified substrates using surface plasmon resonance.
Supported bilayer membranes (SBMs) formed on solid substrates, in particular glass, provide an ideal cell mimicking model system that has been found to be highly useful for biosensing applications. Although the stability of the membrane structures is known to determine the applicability, the subject has not been extensively investigated, largely because of the lack of convenient methods to monitor changes of membrane properties on glass in real time. This work reports the evaluation of the stability properties of a series of SBMs against chemical and air damage by use of surface plasmon resonance spectroscopy and nanoglassified gold substrates. Seven SBMs composed of phosphatidylcholine and DOPC+, including single-component, mixed, protein-reinforced SBMs (rSBMs) and protein-tethered bilayer membranes (ptBLMs), are studied. The stability properties under various conditions, especially the effects of surfactants, organic solvents, and dehydration damage on the bilayers, are compared. PC membranes are found to be easily removed from the glassy surfaces using relatively low concentrations of the surfactants, while DOPC+ is markedly more stable toward nonionic surfactant. DOPC+ membranes also demonstrated remarkable air stability while PC films exhibited considerable damage from dehydration. Doping of cholesterol does not improve PC's stability against SDS and Triton but changes the lipid membrane packing enough to protect against dehydration damage. Although rSBMs and ptBLMs improve air stability to a certain degree, they are still quite susceptible to significant damage/removal from ionic and nonionic surfactants at lower concentrations. Overall, DOPC+ has noted higher stability on glass, likely due to the favorable electrostatic interaction between the silicate surface and the lipid headgroup, making it a good candidate for application. Nanoglassy SPR proves to be an attractive platform capable of rapidly screening film stability in real-time, providing critical information for future work using supported membranes for sensing applications.


Laser microprobe with fourier transform ion cyclotron resonance mass spectrometer for surface analysis.
Fourier transform ion cyclotron resonance laser microprobe mass spectrometry (FTICR LMMS) uses focused laser irradiation of solids with a spot of 5 microm and a FTICR mass analyzer for local analysis with high mass resolution. A new ion source design has been developed to improve the extraction and transfer of ions generated in an external laser microprobe source. Calculations predicted trapping of ions initially emitted with angles up to 40 degrees and 60 degrees from the surface and from a distance of 1 mm above the sample, respectively. The analytical performances of the method have been verified on two sets of test samples. First, detection of chemisorbed benzotriazole on copper, average of two monolayers, has been shown with less sample consumption than typically required in static secondary ion mass spectrometry with a time-of-flight analyzer. Second, experiments on a thermal plate for offset printing have shown the feasibility of analysis and quantification of dyes embedded in a polymer matrix.


Progress in immunization against Klebsiella infections.
Nosocomial infections with Klebsiella spp. are a leading cause of morbidity and mortality. The ability of Klebsiella spp. readily to colonize hospitalized patients, complications in treatment of infections due to R-factor-acquired antibiotic resistance, and the high mortality rate in certain patient populations, point to the need for immunoprophylactic/immunotherapeutic agents for disease control. The potential for vaccination against Klebsiella spp. is discussed in light of recent developments concerning the pathogenesis of Klebsiella infections as relates to the identification of protective antigens as possible vaccine candidates.


Binding Forces of Cellulose Binding Modules on Cellulosic Nanomaterials.
In this study, the interaction forces between different cellulosic nanomaterials and a protein domain belonging to cellulose binding modules family 1 (CBM1) were investigated at the molecular scale. Cellulose binding modules are protein domains found in carbohydrate active enzymes having an affinity toward cellulosic materials. Here, the binding force of a fusion protein containing a cellulose binding module (CBM1) produced recombinantly in E. coli was quantified on different cellulose nanocrystals immobilized on surfaces. Adhesion of the CBM on cellulose with different degrees of crystallinity as well as on chitin nanocrystals was examined. This study was carried out by single molecule force spectroscopy using an atomic force microscope, which enables the detection of binding force of individual molecules. The study contains a preliminary quantification of the interactions at the molecular level that sheds light on the development of new nanocellulose-based nanocomposites with improved strength and elasticity.


Bioaccumulation and trophic transfer of potentially toxic elements in the pelagic thresher shark Alopias pelagicus in Baja California Sur, Mexico.
Pelagic thresher shark (Alopias pelagicus) is a circumglobal species with high ecological and economic importance. Concentrations of mercury, selenium and cadmium in the muscle and liver tissues of A. pelagicus captured from Baja California Sur, Mexico were determined for assessing the potential human hazard. Results revealed that the average concentrations of Hg (0.76 mg kg-1) and Cd (0.18 mg kg-1) in muscle tissues were below the maximum permissible limits for human consumption. Se in the muscles were relatively low (mean: 0.30 mg kg-1 -1) resulting in a molar excess of Hg over Se. Average levels of hepatic Cd were extremely higher than the maximum limit for consumption. Organotropism of Hg was muscle > liver, whereas Se and Cd presented an order of liver > muscle. Biomagnification Factor (BMF) emphasized the trophic transfer of elements. Selenium Health Benefit value was negative (-3.76) posing potential health risks demanding regular monitoring for health risks.


Molecular cloning of a potential proteinase activated receptor.
A DNA sequence encoding a G-protein-coupled receptor was isolated from a mouse genomic library. The predicted protein is similar in structure to the thrombin receptor and has a similar activation mechanism. When expressed in Xenopus laevis oocytes, the receptor was activated by low concentrations of trypsin (EC 220.127.116.11) and by a peptide (SLIGRL) derived from the receptor sequence, but was not activated by thrombin (EC 184.108.40.206). Trypsin failed to activate a mutant receptor in which the presumed cleavage site Arg-34-Ser-35 was changed to an Arg-Pro sequence. The agonist peptide (SLIGRL) activated equally well mutant and wild-type receptors. Northern blot analysis demonstrated receptor transcripts in highly vascularized tissues such as kidney, small intestine, and stomach. Because this, to our knowledge, is the second example, besides the thrombin receptor, of a proteolytically activated seven-transmembrane G-protein-coupled receptor, we have provisionally named it proteinase activated receptor 2.


Role of spatial inhomogenity in GPCR dimerisation predicted by receptor association-diffusion models.
G protein-coupled receptor (GPCR) association is an emerging paradigm with far reaching implications in the regulation of signalling pathways and therapeutic interventions. Recent super resolution microscopy studies have revealed that receptor dimer steady state exhibits sub-second dynamics. In particular the GPCRs, muscarinic acetylcholine receptor M1 (M1MR) and formyl peptide receptor (FPR), have been demonstrated to exhibit a fast association/dissociation kinetics, independent of ligand binding. In this work, we have developed a spatial kinetic Monte Carlo model to investigate receptor homo-dimerisation at a single receptor resolution. Experimentally measured association/dissociation kinetic parameters and diffusion coefficients were used as inputs to the model. To test the effect of membrane spatial heterogeneity on the simulated steady state, simulations were compared to experimental statistics of dimerisation. In the simplest case the receptors are assumed to be diffusing in a spatially homogeneous environment, while spatial heterogeneity is modelled to result from crowding, membrane micro-domains and cytoskeletal compartmentalisation or 'corrals'. We show that a simple association-diffusion model is sufficient to reproduce M1MR association statistics, but fails to reproduce FPR statistics despite comparable kinetic constants. A parameter sensitivity analysis is required to reproduce the association statistics of FPR. The model reveals the complex interplay between cytoskeletal components and their influence on receptor association kinetics within the features of the membrane landscape. These results constitute an important step towards understanding the factors modulating GPCR organisation.


Functional MRI BOLD response in sickle mice with hyperalgesia.
Patients with sickle cell anemia (SCA) have abnormal hemoglobin (sickle hemoglobin S) leading to the crystallization of hemoglobin chains in red blood cells (RBCs), which assume sickle shape and display reduced flexibility. Sickle RBCs (sRBCs) adhere to vessel walls and block blood flow, thus preventing oxygen delivery to the tissues leading to vaso-occlusive crises (VOC), acute pain and organ damage. SCA patients often have chronic pain that can be attributed to inflammation, vasculopathy, neuropathy, ischemia-reperfusion injury and organ damage. Blood oxygenation level-dependent (BOLD) based functional magnetic resonance imaging (fMRI) technique that is commonly used for noninvasively mapping spontaneous or evoked brain activation in human or animal models has been applied in this study to assess abnormal oxygenation change in the brains of mice with SCA in response to hypoxia. We found that hyperalgesic HbSS-BERK sickle mice with chronic pain display reduced BOLD response to a hypoxia challenge compared to their control HbAA-BERK mice. Hypoxia/reoxygenation (H/R) treated sickle mice under acute pain episode exhibit even smaller BOLD signal changes than sickle mice without H/R, suggestive of correlations between cerebral BOLD signal changes and nociception.


Preparation of lipid nanoemulsions by premix membrane emulsification with disposable materials.
The possibility to prepare nanoemulsions as drug carrier systems on small scale was investigated with disposable materials. For this purpose premix membrane emulsification (premix ME) as a preparation method for nanoemulsions with narrow particle size distributions on small scale was used. The basic principle of premix ME is that the droplets of a coarse pre-emulsion get disrupted by the extrusion through a porous membrane. In order to implement the common preparation setup for premix ME with disposable materials, the suitability of different syringe filters (made from polyethersulfone, cellulose acetate, cellulose ester and nylon) and different pharmaceutically relevant emulsifiers (phospholipids, polysorbate 80 and sucrose laurate) for the preparation of nanoemulsions was investigated. Already the preparation of the premix could be realized by emulsification with the help of two disposable syringes. As shown for a phospholipid-stabilized emulsion, the polyethersulfone filter was the most appropriate one and was used for the study with different emulsifiers. With this syringe filter, the median particle size of all investigated emulsions was below 500nm after 21 extrusion cycles through a 200nm filter and a subsequent extrusion cycle through a 100nm filter. Furthermore, the particle size distribution of the polysorbate 80- and sucrose laurate-stabilized emulsions prepared this way was very narrow (span value of 0.7).


Advancement of Lipid-Based Nanocarriers and Combination Application with Physical Penetration Technique.
On account of the advantages of transdermal delivery and the application situation of transcutaneous technology in transdermal delivery, the article critically comments on nanosystems as permeation enhancement model. Nanosystems possess great potential for transcutaneous drug delivery. This review focuses on recent advances in lipid-based nanocarriers, including liposome, transfersomes, ethosomes, nanoemulsions, solid lipid nanoparticles, nanostructured lipid carriers and combination application of the lipid-based nanocarriers with microneedle, iontophoresis, electroporation and sonophoresis in the field for the development of the transdermal drug delivery system. We attempted to give an overview of lipid-based nanocarriers with the aim to improve transdermal and dermal drug delivery. A special focus is given to the nanocarrier composition, characteristic and interaction mechanisms through the skin. Recent combination applications of lipid-based nanocarriers with the physical penetration technology demonstrate the superiority of the combined use of nanocarriers and physical methods in drug penetration enhancement compared to their single use. In the future, lipidbased nanocarriers will play a greater role in the field of transdermal and dermal drug delivery.


Late outcomes for the surgical management of absent pulmonary valve syndrome in infants.
Absent pulmonary valve syndrome (APVS) is a rare cardiac malformation that is usually associated with aneurysmal dilatation of pulmonary arteries and respiratory distress. The surgical mortality of neonates and infants with APVS has decreased tremendously, from 60% in 1980s to 10-20% recently. This study retrospectively reviews surgical outcomes of our 10-year experience in patients with APVS. From 2002 to 2012, 42 patients with APVS underwent surgical correction. Thirty-seven patients had APVS as a variant of tetralogy of Fallot, 4 with double outlet right ventricle and 1 with ventricular septal defect. Respiratory distress was present in 12 infants. Four patients needed continuous positive airway pressure and 5 required intubation with mechanical ventilation before surgery. There was no hospital death and 3 late deaths. The mean follow-up time was 62.71 ± 34.31 months. Significant differences were found in the duration of postoperative ventilation between patients with or without respiratory distress (P = 0.009) and patients with left or right aortic arch (P = 0.012). The Kaplan-Meier curve indicated that overall survival at 5 and 10 years was 92.4%. The survival rates between patients with or without respiratory distress were 72.7 and 100%, respectively (P = 0.003). Overall mortality was associated with longer cardiopulmonary bypass time (P = 0.004) and lower weight at operation (P = 0.042). There were no significant differences in survival and postoperative data such as the duration of ventilation or intensive care unit stay and New York Heart Association class among the three methods of right ventricular outflow tract (RVOT) reconstruction. Surgical treatment of APVS has got favourable outcomes in terms of mortality and reoperation rate. Different methods of RVOT reconstruction do not affect the surgical outcome. Patients required long-term follow-up for postoperative respiratory complications secondary to persistent bronchomalacia.


Molecular Mechanism of Uptake of Cationic Photoantimicrobial Phthalocyanine across Bacterial Membranes Revealed by Molecular Dynamics Simulations.
Phthalocyanines are aromatic macrocyclic compounds, which are structurally related to porphyrins. In clinical practice, phthalocyanines are used in fluorescence imaging and photodynamic therapy of cancer and noncancer lesions. Certain forms of the substituted polycationic metallophthalocyanines have been previously shown to be active in photodynamic inactivation of both Gram-negative and Gram-positive bacteria; one of them is zinc octakis(cholinyl)phthalocyanine (ZnPcChol8+). However, the molecular details of how these compounds translocate across bacterial membranes still remain unclear. In the present work, we have developed a coarse-grained (CG) molecular model of ZnPcChol8+ within the framework of the popular MARTINI CG force field. The obtained model was used to probe the solvation behavior of phthalocyanine molecules, which agreed with experimental results. Subsequently, it was used to investigate the molecular details of interactions between phthalocyanines and membranes of various compositions. The results demonstrate that ZnPcChol8+ has high affinity to both the inner and the outer model membranes of Gram-negative bacteria, although this species does not show noticeable affinity to the 1-palmitoyl-2-oleoyl- sn-glycero-3-phosphatidylcholine membrane. Furthermore, we found out that the process of ZnPcChol8+ penetration toward the center of the outer bacterial membrane is energetically favorable and leads to its overall disturbance and formation of the aqueous pore. Such intramembrane localization of ZnPcChol8+ suggests their twofold cytotoxic effect on bacterial cells: (1) via induction of lipid peroxidation by enhanced production of reactive oxygen species (i.e., photodynamic toxicity); (2) via rendering the bacterial membrane more permeable for additional Pc molecules as well as other compounds. We also found that the kinetics of penetration depends on the presence of phospholipid defects in the lipopolysaccharide leaflet of the outer membrane and the type of counterions, which stabilize it. Thus, the results of our simulations provide a detailed molecular view of ZnPcChol8+ "self-promoted uptake", the pathway previously proposed for some small molecules crossing the outer bacterial membrane.


The impact of substrate surface defects on the properties of two-dimensional van der Waals heterostructures.
The recent emergence of vertically stacked van der Waals (vdW) heterostructures provides new opportunities for these materials to be employed in a wide range of novel applications. Understanding the interlayer coupling in the stacking geometries of the heterostructures and its effect on the resultant material properties is particularly important for obtaining materials with desirable properties. Here, we report that the atomic bonding between stacked layers and thereby the interlayer properties of the vdW heterostructures can be well tuned by the substrate surface defects using WS2 flakes directly grown on graphene. We show that the defects of graphene have no significant effect on the crystal structure or the quality of the grown WS2 flakes; however, they have a strong influence on the interlayer interactions between stacked layers, thus affecting the layer deformability, thermal stability, and physical and electrical properties. Our experimental and computational investigations also reveal that WS2 flakes grown on graphene defects form covalent bonds with the underlying graphene via W atomic bridges (i.e., formation of larger overlapping hybrid orbitals), enabling these flakes to exhibit different intrinsic properties, such as higher conductivity and improved contact characteristics than heterostructures that have vdW interactions with graphene. This result emphasizes the importance of understanding the interlayer coupling in the stacking geometries and its correlation effect for designing desirable properties.


Atrial pacing: who do we pace and what do we expect? Experiences with 100 atrial pacemakers.
The records of 100 patients with permanent atrial pacemakers implanted over a 7-year period were reviewed to assess the role and results of this mode of pacing. Indications for pacing were sick sinus syndrome in 91, carotid sinus hypersensitivity in 3, and use of an antitachycardia device in 6 patients. The mean follow-up period was 32.9 months. Symptomatic relief was good. Lead dislodgment occurred in 11 patients (usually in the first week). Threshold rises not amenable to reprogramming occurred in three patients and loss of sensing occurred in seven patients but only one required intervention. Overall, 21 patients required reoperation. The type of lead did not influence the need for reoperation that appeared to be related to the experience of the operator. Complete atrioventricular block occurred in three patients, two of whom had carotid sinus hypersensitivity and one had sick sinus syndrome. Chronic atrial fibrillation occurred in five patients, none of whom required revision of the pacemaker system. Atrial pacing is a satisfactory pacing mode in patients with sick sinus syndrome. Provided satisfactory atrioventricular conduction has been shown by incremental atrial pacing to at least 120 beats/min and carotid hypersensitivity is absent, progression to complete atrioventricular block is uncommon. Greater implanting skills are required for good results.


Cyclic AMP decreases the expression of a neuronal marker (GAD67) and increases the expression of an astroglial marker (GFAP) in C6 cells.
C6 cells express proteins and mRNAs that are characteristic of both glia and neurons. Agents that increase intracellular levels of cyclic AMP (cAMP) decrease the enzymatic activity of glutamate decarboxylase (GAD), a neuronal marker, and the mRNA levels for one of the two GAD isoenzymes, GAD67. This reduction is accompanied by increased levels of glial fibrillary acidic protein (GFAP) mRNA, an astrocyte marker. Transient transfection assays, in which a 2-kb upstream regulatory region of the human GFAP gene was linked to a reporter gene, indicate that at least some of the cAMP-mediated increase of GFAP mRNA levels is due to increased transcription. Increases in intracellular cAMP appear to induce differentiation of C6 cells toward a more mature astrocyte phenotype.


Caregiver burden: historical development.
Within the last decade there has been a multitude of research related to the phenomenon of caregiver burden. Although the concept is relatively new in the literature, the responsibility and ultimately consequences of caring for another family member has existed for centuries. Often this responsibility lies with the female members of the family, but lately more husbands, sons, close family members, or friends have assumed the primary caregiver role. This article reviews the concept of caregiver burden from a historical perspective. Research related to the definition of the concept and its impact on women, men, and siblings and the effect of ethnicity, is discussed.


Mechanized toxicological serum tests in screening hospitalized patients.
A spectrum of quantitative and qualitative methods was adapted to the RA-1000/RA-XT selective analyser for the purpose of excluding or detecting common types of intoxication in the emergency laboratory of our primary care community hospital. Ethanol and salicylates (measured photometrically) and acetaminophen (measured immunologically by EMIT tox) were quantitatively analysed in serum. immunological group tests (EMIT tox) for barbiturates, benzodiazepines, tricyclic antidepressants and related compounds were used for qualitative analysis. Well established clinical chemical methods (aspartarte aminotransferase, alanine aminotransferase, creatine kinase, pseudocholinesterase, glucose and lactate) were applied to the serum samples using the same selective analyser. Within and between run precision, accuracy, recovery and detection ranges (linearity) fulfilled the recommendations of forefield toxicological analysis for all methods. Ethanol (g/l), measured photometrically with the RA-1000 analyser, agreed with the reference method (headspace gas-chromatography) with a correlation coefficient greater than 0.99 (y = 0.06 + 0.98x). Acetaminophen and salicylates showed correlation coefficients greater than 0.94 and greater than 0.99, when compared with manual colorimetric procedures (acetaminophen (mg/l): y = -3.22 + 0.896x; salicylates (mg/l): y = -2.1 + 1x). Qualitative group tests for barbiturates, benzodiazepines and tricyclic antidepressants measured with the RA-1000 analyser were in good agreement with the EMIT single test procedure. The ranges of the quantitative methods allowed quantification of analytes from therapeutic (non-toxic) to very high levels in undiluted samples (ethanol 0.05 up to 4 g/l; salicylates 32 up to 1200 mg/l and acetaminophen 1.9 up to 200 mg/l). The low detection limits of the qualitative tests allowed the recognition of compounds in plasma that were present in low concentrations and/or displayed only minor reactivity with the antibodies provided by the EMIT tox test kits. As a consequence, decision limits for all three group tests in serum were lowered to near the detection limit: (table: see text) For quantitative tests the lower limits of quantification were: (table: see text) The working reagents were stable for at least 14 days at 4-8 degrees C. Calibration curves were stable over the expiration period of reconstituted original reagents (6-12 weeks), also when working reagents were prepared in aliquots from stored reconstituted reagents. Application of the newly adapted programme to serum samples of nearly two hundred patients showed it to be suitable for screening patients in which intoxication is suspected or needs to be excluded.


A phase I study of UFT/leucovorin, carboplatin, and paclitaxel in combination with external beam radiation therapy for advanced esophageal carcinoma.
Concurrent chemotherapy and radiation therapy (RT) are used to treat patients with esophageal cancer. The optimal combination of chemotherapeutic agents with RT is not well established. We evaluated the safety and preliminary efficacy of a combination of UFT/leucovorin, carboplatin, and paclitaxel with RT in a Phase I study of patients with advanced esophageal cancer. Patients with squamous cell carcinoma or adenocarcinoma of the esophagus initially received UFT/leucovorin, carboplatin, and paclitaxel with RT (1.8 Gy daily to 45 Gy). After completion, the disease was restaged and patients were evaluated for surgery. Primary end points included determination of dose-limiting toxicities (DLTs) and a recommended Phase II dose. Secondary objectives included determination of non-DLTs, as well as preliminary radiographic and pathologic response rates. Twelve patients were enrolled (11 men, 1 woman). All were assessable for toxicity and efficacy. One of 6 patients at Dose Level 1 (UFT/leucovorin, 200/30 mg twice daily on RT days; carboplatin, area under the curve [AUC] 5, Weeks 1 and 4; paclitaxel, 175 mg/m2 Weeks 1 and 4) had a DLT (febrile neutropenia). Of these 6 patients, 4 underwent esophagectomy and none achieved a pathologic complete response. Six patients were then enrolled at Dose Level 2 (UFT/leucovorin, 300/30 mg in the morning and 200/30 mg in the evening on RT days; carboplatin, AUC 5, Weeks 1 and 4; paclitaxel, 175 mg/m2 Weeks 1 and 4). Two of 6 patients at Dose Level 2 developed DLTs (febrile neutropenia in both). Esophagectomy was performed in 3 patients, with 2 achieving a pathologic complete response. Maximum tolerated doses in this study were UFT/leucovorin, 200/30 mg twice daily on RT days; carboplatin, AUC 5, Weeks 1 and 4; and paclitaxel, 175 mg/m2 Weeks 1 and 4 when delivered with external RT. In this small study, this regimen appears active, but toxic.


Comparative study of DNA adduct formation in mice following inhalation of smoke from cigarettes that burn or primarily heat tobacco.
The genotoxic potential of mainstream smoke from a test cigarette (TOB-HT) that primarily heats tobacco and a representative tobacco-burning cigarette (Kentucky reference 1R4F) was compared in male B6C3/F1 mice after nose-only inhalation exposure. Mice were exposed 1 hr per day, 5 days/ week for a 4 week period to mainstream smoke at concentrations of 0, 0.16, 0.32, and 0.64 mg total particulate matter/liter of air. Micronuclei formation in bone marrow and peripheral blood polychromatic erythrocytes (PCE) of animals exposed to either the TOB-HT or 1R4F cigarette was not significantly different compared with control animals exposed nose-only to filtered and humidified air (sham controls). DNA adduct measurement by the 32P-post-labeling method indicated an exposure-dependent increase in lung adducts of animals exposed to 1R4F cigarette smoke at all three concentrations with the mid and high exposure groups exhibiting statistically significant increases (P < 0.05) in adduct formation compared to sham-exposed animals. The concentration of DNA adducts in the lungs of animals exposed to the TOB-HT cigarette was not significantly increased (P < 0.05) at any concentration compared to sham-exposed controls. A statistically significant (P < 0.05) concentration-dependent formation of DNA adducts was also observed in the heart tissues of animals exposed to smoke from the 1R4F cigarette at all three concentrations, but no significant increase in adduct formation was observed in heart DNA of the animals exposed to the TOB-HT cigarette (P < 0.05). Under the conditions of this experiment, the mainstream smoke from the TOB-HT cigarette was demonstrated to be less genotoxic in mice than mainstream smoke from the 1R4F cigarette, which is representative of cigarettes in the current U.S. market.


Comparative-genomic analysis reveals dynamic NLR gene loss and gain across Apiaceae species.
Introduction: Nucleotide-binding leucine-rich repeat (NLR) genes play a crucial role in green plants' responding to various pathogens. Genome-scale evolutionary studies of NLR genes are important for discovering and applying functional NLR genes. However, little is known about the evolution of NLR genes in the Apiaceae family including agricultural and medical plants. Methods: In this study, comparative genomic analysis was performed in four Apiaceae species to trace the dynamic evolutionary patterns of NLR genes during speciation in this family. Results: The results revealed different number of NLR genes in these four Apiaceae species, namely, Angelica sinensis (95), Coriandrum sativum (183), Apium graveolens (153) and Daucus carota (149). Phylogenetic analysis demonstrated that NLR genes in these four species were derived from 183 ancestral NLR lineages and experienced different levels of gene-loss and gain events. The contraction pattern of the ancestral NLR lineages was discovered during the evolution of D. carota, whereas a different pattern of contraction after first expansion of NLR genes was observed for A. sinensis, C. sativum and A. graveolens. Discussion: Taken together, rapid and dynamic gene content variation has shaped evolutionary history of NLR genes in Apiaceae species.


Phylogenetic relationships among cetaceans revealed by Y-chromosome sequences.
The Y chromosome has recently come into the spotlight as a new and efficient genetic marker for tracing paternal lineages. We reconstructed cetacean phylogeny using a 1.7-kbp fragment of the non-recombining Y chromosome (NRY), including the SRY gene and a flanking non-coding region. The topology of the Y-chromosome tree is robust to various methods of analysis and exhibits high branch-support values, possibly due to the absence of recombination, small effective population size, and low homoplasy. The Y-chromosome tree indicates monophyly of each suborder, Mysticeti and Odontoceti, with high branch support values (BS> or =86%; PP> or =98%). In the Odontoceti clade, three superfamilies, Physeteroidea, Ziphioidea, and Delphinoidea, diverged soon after the split between Mysticeti and Odontoceti. Our analysis allows resolution of this rapid radiation and indicates that Physeteroidea is basal in the Odontoceti clade (BS, 99%; PP, 100%; MBS, 61%). The major split within the superfamily Delphinoidea is between the Delphinidae clade and the Monodontidae+ Phocoenidae clade. The phylogenetic relationships among delphinid species are ambiguous, probably because of the rapid radiation of this family. In the Mysticeti clade, the first major split is between Balaenidae and Balaenopteridae; within Balaenopteridae, a Balaenoptera acutorostrata+B. bonaerensis (minke whales) clade forms a sister clade with the other balaenopterid species. Megaptera novaeangliae is nested within Balaenoptera, making the latter paraphyletic. The low homoplasy exhibited by the Y-chromosome data presented here suggests that an extended data set incorporating longer sequences would provide better resolution of cetacean lower-level pylogeny.


Impact of sarcopenia on the outcomes of elective total arch replacement in the elderly†.
The purpose of this study was to identify the cut-off value of sarcopenia based on the psoas muscle area index and evaluate early and late outcomes following elective total arch replacement in the elderly. Sarcopenia was assessed by the psoas muscle area index [defined as the psoas muscle area at the L3 level on computed tomography (cm 2 )/body surface area (m 2 )]. The cut-off value for sarcopenia was defined as > 2 standard deviations below the mean psoas muscle area index value obtained from 464 normal control patients. Between October 1999 and July 2015, 266 patients who were ≥ 65 years and had undergone psoas muscle area index measurement underwent elective total arch replacement. These patients were classified into the sarcopenia (Group S, n = 81) and non-sarcopenia (Group N, n = 185) groups. The mean age was 76.2 ± 5.6 years in Group S and 75.7 ± 5.7 years in Group N ( P = 0.553). Hospital mortality was 3.7% (3/81) in Group S and 2.2% (4/185) in Group N ( P = 0.483). Mean follow-up was 48.3 ± 38.7 months. Five-year survival was significantly worse in Group S (S: 63.2 ± 6.6% vs N: 88.7 ± 2.6%, P < 0.001). A multivariable Cox proportional hazard analysis showed that sarcopenia significantly predicted poor survival (hazard ratio 2.59; 95% confidence interval 1.27-5.29; P = 0.011). Sarcopenia did not predict hospital death following total arch replacement, but it was negatively associated with overall survival. Sarcopenia can be an additional risk factor to estimate the outcomes of thoracic aortic surgery.


Pesticide residues in nectar and pollen of melon crops: Risk to pollinators and effects of a specific pesticide mixture on Bombus terrestris (Hymenoptera: Apidae) micro-colonies.
Residues detected in pollen collected by honey bees are often used to estimate pesticide exposure in ecotoxicological studies. However, for a more accurate assessment of pesticides effect on foraging pollinators, residues found directly on flowers are a more realistic exposure approximation. We conducted a multi-residue analysis of pesticides on pollen and nectar of melon flowers collected from five fields. The cumulative chronic oral exposure Risk Index (RI) was calculated for Apis mellifera, Bombus terrestris and Osmia bicornis to multiple pesticides. However, this index could underestimate the risk since sublethal or synergistic effects are not considered. Therefore, a mixture containing three of the most frequently detected pesticides in our study was tested for synergistic impact on B. terrestris micro-colonies through a chronic oral toxicity test. According to the result, pollen and nectar samples contained numerous pesticide residues, including nine insecticides, nine fungicides, and one herbicide. Eleven of those were not applied by farmers during the crop season, revealing that melon agroecosystems may be pesticide contaminated environments. The primary contributor to the chronic RI was imidacloprid and O. bircornis is at greatest risk for lethality resulting from chronic oral exposure at these sites. In the bumblebee micro-colony bioassay, dietary exposure to acetamiprid, chlorpyrifos and oxamyl at residue level concentration, showed no effects on worker mortality, drone production or drone size and no synergies were detected when pesticide mixtures were evaluated. In conclusion, our findings have significant implications for improving pesticide risk assessment schemes to guarantee pollinator conservation. In particular, bee pesticide risk assessment should not be limited to acute exposure effects to isolated active ingredients in honey bees. Instead, risk assessments should consider the long-term pesticide exposure effects in both pollen and nectar on a range of bees that reflect the diversity of natural ecosystems and the synergistic potential among pesticide formulations.


Immunoproteomic identification of MbovP579, a promising diagnostic biomarker for serological detection of Mycoplasma bovis infection.
A lack of knowledge regarding the antigenic properties of Mycoplasma bovis proteins prevents the effective control of bovine infections using immunological approaches. In this study, we detected and characterized a specific and sensitive M. bovis diagnostic biomarker. After M. bovis total proteins and membrane fractions were separated with two dimensional gel electrophoresis, proteins reacting with antiserawere detected using MALDI-TOF MS. Thirty-nine proteins were identified, 32 of which were previously unreported. Among them, immunoinformatics predicted eight antigens, encoded by Mbov_0106, 0116, 0126, 0212, 0275, 0579, 0739, and 0789, to have high immunological value. These genes were expressed in E. coli after mutagenesis of UGA to UGG using overlap extension PCR. A lipoprotein, MbovP579, encoded by a functionally unknown gene, was a sensitive and specific antigen for detection of antibodies in sera from both M. bovis-infected and vaccinated cattle. The specificity of MbovP579 was confirmed by its lack of cross-reactivity with other mycoplasmas, including Mycoplasma agalactiae. An iELISA based on rMbovP579 detected seroconversion 7 days post-infection (dpi). The ELISA had sensitivity of 90.2% (95% CI: 83.7%, 94.3%) and a specificity of 97.8% (95% CI: 88.7%, 99.6%) with clinical samples. Additional comparative studies showed that both diagnostic and analytic sensitivities of the ELISA were higher than those of a commercially available kit (p<0.01). We have thus detected and characterized the novel antigen, MbovP579, and established an rMbovP579-based ELISA as a highly sensitive and specific method for the early diagnosis of M. bovis infection.


Right-sided aortic arch with the retroesophageal left subclavian artery as the fourth branch.
We present a rare variation of the right-sided aortic arch with the retroesophageal left subclavian artery as the forth branch found in a cadaver of an 89-year-old Korean woman during a routine dissection. In this case, the first branch that arose from the ascending aorta was the left common carotid artery, which crossed ventral to the trachea in a left cephalic direction, followed by the right common carotid artery and then the right subclavian artery. Distal to these branches the aortic arch ran dorsally, passing between the esophagus and the vertebra. The left subclavian artery arose from the descending portion of the aortic arch, crossing over to the left upper extremity behind the esophagus. This anomaly was not accompanied by congenital heart disease. Accurate information regarding this variation is of great importance to surgeons for its early identification and preservation during interventions and to radiologists for precise interpretation of angiograms.


Pathophysiology of the Gut and the Microbiome in the Host Response.
To describe and summarize the data supporting the gut as the motor driving critical illness and multiple organ dysfunction syndrome presented at the National Institute of Child Health and Human Development MODS Workshop (March 26-27, 2015). Summary of workshop keynote presentation. Not applicable. Presented by an expert in the field, the data assessing the role of gastrointestinal dysfunction driving critical illness were described with a focus on identifying knowledge gaps and research priorities. Summary of presentation and discussion supported and supplemented by relevant literature. The understanding of gut dysfunction in critical illness has evolved greatly over time, and the gut is now often considered as the "motor" of critical illness. The association of the gut with critical illness is supported by both animal models and clinical studies. Initially, the association between gut dysfunction and critical illness focused primarily on bacterial translocation into the bloodstream. However, that work has evolved to include other gut-derived products causing distant injury via other routes (e.g., lymphatics). Additionally, alterations in the gut epithelium may be associated with critical illness and influence outcomes. Gut epithelial apoptosis, intestinal hyperpermeability, and perturbations in the intestinal mucus layer have all been associated with critical illness. Finally, there is growing evidence that the intestinal microbiome plays a crucial role in mediating pathology in critical illness. Further research is needed to better understand the role of each of these mechanisms and their contribution to multiple organ dysfunction syndrome in children.


Oxytocin in maternal and fetal blood.
Radioimmunoassayable plasma oxytocin (OT) has been measured in maternal and fetal blood. Simultaneous samples were obtained in maternal forearm venous blood and in umbilical venous and arterial blood in 29 patients at term delivery. In addition, maternal forearm venous blood samples were also obtained 10 minutes prior to delivery. Mean OT level in maternal plasma at delivery was 82 +/- 12 muU/ml, and at 10 minutes prior to delivery the mean OT level was 90 +/- 11 muU/ml. The umbilical arterial plasma OT showed 95 +/- 12 muU/ml and the umbilical vein plasma OT was 60 +/- 10 muU/ml. Oxytocin levels higher in maternal blood than in fetal blood were found with the following incidence: In 51% of samples there was more OT in maternal venous blood than in umbilical arterial blood, and in 84% of samples there was more OT in maternal blood than umbilical vein blood. During the postpartum period, the mean maternal plasma OT was 66 +/- 8 muU/ml for the first day, and 50 +/- 9 muU/ml and 54 +/- 9 muU/ml for the second and third days, respectively. This study indicates that both the fetus and the mother are active producers of oxytocin.


[Follicular lymphoma: recent advances].
Follicular lymphoma (FL) is a neoplasm comprising germinal center B cells, which typically has, at least, a partially follicular pattern. Although the natural course of the disease is indolent, the chemosensitivity is low, which results in frequent relapse and immunosuppression. Therefore, the tumor stage and tumor burden are essential criteria for deciding on the treatment strategy. For advanced-stage patients with a low tumor burden, watchful waiting is a standard option. The optimal timing for administrating rituximab is a critical clinical question. For advanced-stage patients with a high tumor burden, R-CHOP, R-CVP, and BR are the standard options. Recently, the second-generation CD20 antibody was added to the options. Several new agents are now under development for relapsed FL.


Hydroxylated and methoxylated polybrominated diphenyl ethers in long-tailed ducks (Clangula hyemalis) and their main food, Baltic blue mussels (Mytilus trossulus × Mytilus edulis).
Long-tailed ducks (Clangula hyemalis) that breed in northern Europe and western Siberia and commonly winter in the Baltic Sea, are threatened by a significant population decrease. The ducks are, by primarily feeding on Baltic blue mussels (Mytilus trossulus × Mytilus edulis) while wintering in the Baltic Sea, potentially subjected to high levels of toxic hydroxylated polybrominated diphenyl ethers (OH-PBDEs). To assess long-tailed ducks exposure to polybrominated phenols (PBPs), polybrominated anisoles (PBAs), hydroxylated polybrominated diphenyl ethers (OH-PBDEs), their methylated counterparts (MeO-PBDEs) and polybrominated diphenyl ethers (PBDEs), livers of ten long-tailed ducks wintering in the Baltic Sea were analysed. Pattern and levels of analytes in long-tailed ducks (liver) and blue mussels sampled in March and May at nine sites in the Baltic Sea were compared. The geometric mean concentration (ng/g l.w.) in livers of long-tailed ducks and Baltic blue mussels were: Σ(2)PBPs: 0.57 and 48; Σ(2)PBAs: 0.83 and 11; Σ(7)OH-PBDEs: 6.1 and 45; Σ(7)MeO-PBDEs: 3.8 and 69; Σ(7)PBDEs: 8.0 and 7.2, respectively. Based on an estimated daily intake of 450 g fresh blue mussel meat, long-tailed ducks daily dietary intake of brominated substances while foraging in the Baltic Sea in March-May was estimated to; 390 ng Σ(2)PBPs, 90 ng Σ(2)PBAs, 370 ng Σ(7)OH-PBDEs, 590 ng Σ(7)MeO-PBDEs and 59 ng Σ(7)PBDEs. The low levels of PBPs, PBAs, OH-PBDEs and MeO-PBDEs in the long-tailed duck livers compared to blue mussel, despite a continuous daily intake, suggest that these compounds are poorly retained in long-tailed ducks.


Sequencing and phylogenetic analysis of tobacco virus 2, a polerovirus from Nicotiana tabacum.
The complete genome sequence of a new virus, provisionally named tobacco virus 2 (TV2), was determined and identified from leaves of tobacco (Nicotiana tabacum) exhibiting leaf mosaic, yellowing, and deformity, in Anhui Province, China. The genome sequence of TV2 comprises 5,979 nucleotides, with 87% nucleotide sequence identity to potato leafroll virus (PLRV). Its genome organization is similar to that of PLRV, containing six open reading frames (ORFs) that potentially encode proteins with putative functions in cell-to-cell movement and suppression of RNA silencing. Phylogenetic analysis of the nucleotide sequence placed TV2 alongside members of the genus Polerovirus in the family Luteoviridae. To the best our knowledge, this study is the first report of a complete genome sequence of a new polerovirus identified in tobacco.


[The eye department at Copenhagen municipal hospital. Part 3: the epoch of Braendstrup, 1957-1980].
Poul Braendstrup (PB 1912-80) was the best qualified to become head of the eye department at Kommunehospitalet (KH) in 1957, with an eminent great scientific production. He continued this work all his life. Three assistant doctors with different specialities were appointed 1966-72: M. Norn (external eye), K. Norskov (strabism) and Sv. E. Lorentzen (retina). Reasons for the ideal cooperation in the department are described (staff-meetings, christmas-staff-arrangements, meetings, courses etc). The number of operations increased steadily in the period, especially cataract-extractions. The newest techniques and instruments were used (cornea-grafting, vitreous surgery, xenoncoagulation (PB study tour in 1962) before laser-treatment etc). The scientific production increases, surpassing Rigshospitalet after 1967. The education was effective at all levels: In the eye-department, doctors evenings, A-and B-courses for eye specialists to become, referring eye specialists, students (from 1972 as an independent part (sect IV) of the University of Copenhagen with an ordinary professor from 1975). It was disturbing with the many removals at KH and at last the final total removal to KKHH (Hvidovre), causing loss of some of the best staff members, instrument breaks, increasing waiting lists up to one year, inconvenience for the patients and many other difficulties. This could be a warning against the continued great movings of departments in Copenhagen (H:S). The administration is criticised. PB's rationalisation of the duty-plan for all Copenhagen eye departments already from 1966 independent of the administration is mentioned.


Structural snapshots of the catalytic cycle of the phosphodiesterase Autotaxin.
Autotaxin (ATX) is a secreted phosphodiesterase that produces the signalling lipid lysophosphatidic acid (LPA). The bimetallic active site of ATX is structurally related to the alkaline phosphatase superfamily. Here, we present a new crystal structure of ATX in complex with orthovanadate (ATX-VO5), which binds the Oγ nucleophile of Thr209 and adopts a trigonal bipyramidal conformation, following the nucleophile attack onto the substrate. We have now a portfolio of ATX structures we discuss as intermediates of the catalytic mechanism: the new ATX-VO5 structure; a unique structure where the nucleophile Thr209 is phosphorylated (ATX-pThr). Comparing these to a complex with the LPA product (ATX-LPA) and with a complex with a phosphate ion (ATX-PO4), that represent the Michaelis complex of the reaction, we observe movements of Thr209, changes in the relative displacement of the zinc ions, and a water molecule that likely fulfils the second nucleophilic attack. We propose that ATX follows the associative two-step in-line displacement mechanism.


Entrapment of human leukocyte interferon in the aqueous interstices of liposomes.
Human leukocyte interferon has been trapped in the aqueous interstices of multilamellar liposomes (phosphatidylcholine, dicetyl phosphate, and cholesterol [7:2:1]). Such liposomes trapped [3H]inulin (aqueous space marker) and interferon to the extent of 0.22 +/- 0.01 mg (n = 8) and 350 +/- 54 U (n = 4) per mumol of liquid, respectively, as judged by molecular sieve chromatography. Interferon trapped within liposomes was resistant to tryptic digestion under conditions which completely inactivated free interferon. Studies in which interferon was added to preformed liposomes excluded the possibility that interferon bound nonspecifically to the outer layer of the multilamellar liposomes. When interferon was added to the aqueous medium in which liposomes of various net surface charges were permitted to form, trapping of interferon varied directly with the interlamellar aqueous compartments of the liposomes. The demonstration that stable liposomes can entrap interferon suggests that these may constitute suitable vectors for the delivery of interferon to cells.