Optimizing immobilization on two-dimensional carboxyl surface: pH dependence of antibody orientation and antigen binding capacity.
The performance of immunosensors is highly dependent on the amount of immobilized antibodies and their remaining antigen binding capacity. In this work, a method for immobilization of antibodies on a two-dimensional carboxyl surface has been optimized using quartz crystal microbalance biosensors. We show that successful immobilization is highly dependent on surface pK(a), antibody pI, and pH of immobilization buffer. By the use of EDC/sulfo-NHS (1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride/N-hydroxysulfosuccinimide) activation reagents, the effect of the intrinsic surface pK(a) is avoided and immobilization at very low pH is therefore possible, and this is important for immobilization of acidic proteins. Antigen binding capacity as a function of immobilization pH was studied. In most cases, the antigen binding capacity followed the immobilization response. However, the antigen-to-antibody binding ratio differed between the antibodies investigated, and for one of the antibodies the antigen binding capacity was significantly lower than expected from immobilization in a certain pH range. Tests with anti-Fc and anti-Fab(2) antibodies on different antibody surfaces indicated that the orientation of the antibodies on the surface had a profound effect on the antigen binding capacity of the immobilized antibodies.


Seminaphthofluorones are a family of water-soluble, low molecular weight, NIR-emitting fluorophores.
A readily accessible new class of near infrared (NIR) molecular probes has been synthesized and evaluated. Specific fluorophores in this unique xanthene based regioisomeric seminaphthofluorone dye series exhibit a combination of desirable characteristics including (i) low molecular weight (339 amu), (ii) aqueous solubility, and (iii) dual excitation and emission from their fluorescent neutral and anionic forms. Importantly, systematic changes in the regiochemistry of benzannulation and the ionizable moieties afford (iv) tunable deep-red to NIR emission from anionic species and (v) enhanced Stokes shifts. Anionic SNAFR-6, exhibiting an unusually large Stokes shift of approximately 200 nm (5,014 cm(-1)) in aqueous buffer, embodies an unprecedented fluorophore that emits NIR fluorescence when excited in the blue/green wavelength region. The successful use of SNAFR-6 in cellular imaging studies demonstrates proof-of-concept that this class of dyes possesses photophysical characteristics that allow their use in practical applications. Notably, each of the new fluorophores described is a minimal template structure for evaluation of their basic spectral properties, which may be further functionalized and optimized yielding concomitant improvements in their photophysical properties.


Digital skin necrosis in congenital afibrinogenaemia associated with hepatitis C virus infection, mixed cryoglobulinaemia and anticardiolipin antibodies.
Congenital afibrinogenaemia is a rare genetic disorder transmitted as an autosomal recessive trait and characterized by the complete absence of fibrinogen in the plasma. We report a 41-year-old woman who suffered from congenital afibrinogenaemia and hepatitis C viral infection and presented with ischaemic necrosis and livedo of the toes. Laboratory investigations showed the presence of mixed cryoglobulinaemia and anticardiolipin antibodies. Resolution occurred with plasmapheresis. We discuss the pathophysiology of this unusual condition and review the literature for skin manifestations associated with this rare haemostasis disorder.


Nonalcoholic fatty liver disease and the renin-angiotensin system: Implications for treatment.
Nonalcoholic fatty liver disease (NAFLD) is the commonest liver disease in Western countries. Treatment of NAFLD is currently based on lifestyle measures and no effective pharmacologic treatment is available so far. Emerging evidence, mainly from animal studies, suggests that the renin-angiotensin-aldosterone system may be of major importance in the pathogenesis of NAFLD and indicates that angiotensin-converting enzyme inhibitors (ACE-I) and angiotensin receptor blockers (ARBs) as a potentially useful therapeutic approach. However, data from human studies are limited and contradictory. In addition, there are few randomized controlled trials (RCTs) on the effects of ACE-I or ARB in patients with NAFLD and most data are from retrospective studies, pilot prospective studies and post hoc analyses of clinical trials. Accordingly, more and larger RCTs are needed to directly assess the effectiveness of ACE-I and ARBs in NAFLD.


Sorafenib inhibits cell migration and stroma-mediated bortezomib resistance by interfering B-cell receptor signaling and protein translation in mantle cell lymphoma.
We evaluated the antitumoral properties of the multikinase inhibitor sorafenib in mantle cell lymphoma (MCL), an aggressive B lymphoma for which current therapies have shown limited efficacy. Sensitivity to sorafenib was analyzed in MCL cell lines and primary samples in the context of BCR and microenvironment simulation. Sorafenib signaling was characterized by quantitative PCR, Western blotting, immunofluorescence, and protein immunoprecipitation. Migration analysis included flow cytometric counting, actin polymerization assays, and siRNA-mediated knockdown of focal adhesion kinase (FAK). In vivo antitumor effect of sorafenib and bortezomib was analyzed in an MCL xenograft mouse model. Sorafenib rapidly dephosphorylates the BCR-associated kinases, Syk and Lyn, as well as FAK, an Src target involved in focal adhesion. In this line, sorafenib displays strong synergy with the Syk inhibitor, R406. Sorafenib also blocks Mcl-1 and cyclin D1 translation, which promotes an imbalance between pro- and antiapoptotic proteins and facilitates Bax release from cyclin D1, leading to the induction of mitochondrial apoptosis and caspase-dependent and -independent mechanisms. Moreover, sorafenib inhibits MCL cell migration and CXCL12-induced actin polymerization. FAK knockdown partially prevents this inhibitory effect, indicating that FAK is a relevant target of sorafenib. Furthermore, sorafenib enhances the antitumoral activity of bortezomib in an MCL xenograft mouse model as well as overcomes stroma-mediated bortezomib resistance in MCL cells. We show for the first time that sorafenib interferes with BCR signaling, protein translation and modulates the microenvironment prosurvival signals in MCL, suggesting that sorafenib, alone or in combination with bortezomib, may represent a promising approach to treat patients with MCL.


New and emerging treatments for estrogen receptor-positive breast cancer: focus on everolimus.
Management of patients with metastatic hormone receptor-positive breast cancer poses a challenge due to the inevitable development of endocrine resistance. Hormone resistance is associated with a complex interaction of the estrogen receptor with growth factors, transmembrane receptors, and intracellular growth cascades. The PI3K/Akt/mTOR pathway plays a major role in hormone resistance and proliferation of breast cancer. Preclinical and clinical data indicate that inhibitors of human epidermal growth factor receptor-2, epidermal growth factor receptor, insulin-like growth factor-1 receptor, and the mammalian target of rapamycin pathway may act synergistically with hormone therapy to circumvent endocrine resistance. Everolimus is currently approved for combination with exemestane in postmenopausal women with advanced hormone receptor-positive breast cancer. However, we still need to unfold the full potential of targeted agents in the hormone-refractory setting and to identify the subsets of patients who will benefit from combination hormonal therapy using targeted agents.


The use of repetitive transcranial magnetic stimulation (rTMS) and transcranial direct current stimulation (tDCS) to relieve pain.
Chronic pain resulting from injury of the peripheral or central nervous system may be associated with a significant dysfunction of extensive neural networks. Noninvasive stimulation techniques, such as repetitive transcranial magnetic stimulation (rTMS) and transcranial direct current stimulation (tDCS) may be suitable to treat chronic pain as they can act on these networks by modulating neural activities not only in the stimulated area, but also in remote regions that are interconnected to the site of stimulation. Motor cortex was the first cortical target that was proved to be efficacious in chronic pain treatment. At present, significant analgesic effects were also shown to occur after the stimulation of other cortical targets (including prefrontal and parietal areas) in acute provoked pain, chronic neuropathic pain, fibromyalgia, or visceral pain. Therapeutic applications of rTMS in pain syndromes are limited by the short duration of the induced effects, but prolonged pain relief can be obtained by repeating rTMS sessions every day for several weeks. Recent tDCS studies also showed some effects on various types of chronic pain. We review the evidence to date of these two techniques of noninvasive brain stimulation for the treatment of pain.


Localized hotspots drive continental geography of abnormal amphibians on U.S. wildlife refuges.
Amphibians with missing, misshapen, and extra limbs have garnered public and scientific attention for two decades, yet the extent of the phenomenon remains poorly understood. Despite progress in identifying the causes of abnormalities in some regions, a lack of knowledge about their broader spatial distribution and temporal dynamics has hindered efforts to understand their implications for amphibian population declines and environmental quality. To address this data gap, we conducted a nationwide, 10-year assessment of 62,947 amphibians on U.S. National Wildlife Refuges. Analysis of a core dataset of 48,081 individuals revealed that consistent with expected background frequencies, an average of 2% were abnormal, but abnormalities exhibited marked spatial variation with a maximum prevalence of 40%. Variance partitioning analysis demonstrated that factors associated with space (rather than species or year sampled) captured 97% of the variation in abnormalities, and the amount of partitioned variance decreased with increasing spatial scale (from site to refuge to region). Consistent with this, abnormalities occurred in local to regional hotspots, clustering at scales of tens to hundreds of kilometers. We detected such hotspot clusters of high-abnormality sites in the Mississippi River Valley, California, and Alaska. Abnormality frequency was more variable within than outside of hotspot clusters. This is consistent with dynamic phenomena such as disturbance or natural enemies (pathogens or predators), whereas similarity of abnormality frequencies at scales of tens to hundreds of kilometers suggests involvement of factors that are spatially consistent at a regional scale. Our characterization of the spatial and temporal variation inherent in continent-wide amphibian abnormalities demonstrates the disproportionate contribution of local factors in predicting hotspots, and the episodic nature of their occurrence.


Participation of the lone tryptophan residue of rat alpha-foetoprotein in its drug-binding sites. Comparison with rat serum albumin.
The participation in drug binding of the lone tryptophan residue of rat alpha-foetoprotein (alpha-FP) and serum albumin, the two main transport proteins of foetal serum, has been studied by two different techniques. Firstly, the effect on phenylbutazone and warfarin binding of the chemical derivatization of the lone tryptophan residue of both proteins by 2-nitrophenylsulphonyl chloride (NPS) was studied. Secondly, the effect of phenylbutazone binding on the intrinsic fluorescence of the tryptophan residue of rat alpha-FP and albumin was investigated. The specific modification of the proteins by NPS did not affect the binding of warfarin by rat alpha-FP and albumin, but greatly decreased the affinity of the high-affinity sites of rat alpha-FP for phenylbutazone, though the numbers of these sites were not significantly changed. However, for albumin a similar decrease in the affinity constant appeared to be due to the reaction conditions. The spectrofluorimetric studies showed that the lone tryptophan residue of alpha-FP and albumin was quenched by phenylbutazone binding, and the quenching paralleled the fractional saturation of the high-affinity site only in the case of albumin. The effect of phenylbutazone binding on the intrinsic fluorescence of rat alpha-FP indicated that the lone tryptophan residue of this foetal protein is not in the same molecular environment as that of albumin, not participating directly in the high-affinity site for phenylbutazone, and the effect may be via some induced conformational change in rat alpha-FP. These results also confirm our previous suggestion that the high-affinity sites for phenylbutazone and warfarin are different on the rat alpha-FP molecule. The results seem to indicate that this is also the case for albumin, but confirmation is necessary.


Reversible alkene insertion into the Pd-N bond of Pd(II)-sulfonamidates and implications for catalytic amidation reactions.
Alkene insertion into Pd-N bonds is a key step in Pd-catalyzed oxidative amidation of alkenes. A series of well-defined Pd(II)-sulfonamidate complexes have been prepared and shown to react via insertion of a tethered alkene. The Pd-amidate and resulting Pd-alkyl species have been crystallographically characterized. The alkene insertion reaction is found to be reversible, but complete conversion to oxidative amination products is observed in the presence of O(2). Electronic-effect studies reveal that alkene insertion into the Pd-N bond is favored kinetically and thermodynamically with electron-rich amidates.


Use of herbs or vitamin/mineral supplements by patients with cancer in Kayseri, Turkey.
This cross-sectional study was performed to determine the consumption rate, duration, and methods of herb or vitamin/mineral use by patients with cancer and to evaluate the effects of alternative therapies in Kayseri, Turkey. A face-to-face interviews was conducted with 203 patients. Results showed that 32% and 18.2% of the patients used herbs or vitamin/mineral supplements, respectively. Nettle was the most commonly used herb, and iron and vitamin C were the most commonly used supplements. There was a significant difference between herb or vitamin/mineral users and nonusers with regard to educational levels. In conclusion, patients with cancer highly tended to use herb or vitamin/mineral supplements.


Cryptic habitats and cryptic diversity: unexpected patterns of connectivity and phylogeographical breaks in a Mediterranean endemic marine cave mysid.
The marine cave-dwelling mysid Hemimysis margalefi is distributed over the whole Mediterranean Sea, which contrasts with the poor dispersal capabilities of this brooding species. In addition, underwater marine caves are a highly fragmented habitat which further promotes strong genetic structuring, therefore providing highly informative data on the levels of marine population connectivity across biogeographical regions. This study investigates how habitat and geography have shaped the connectivity network of this poor disperser over the entire Mediterranean Sea through the use of several mitochondrial and nuclear markers. Five deeply divergent lineages were observed among H. margalefi populations resulting from deep phylogeographical breaks, some dating back to the Oligo-Miocene. Whether looking at the intralineage or interlineage levels, H. margalefi populations present a high genetic diversity and population structuring. This study suggests that the five distinct lineages observed in H. margalefi actually correspond to as many separate cryptic taxa. The nominal species, H. margalefi sensu stricto, corresponds to the westernmost lineage here surveyed from the Alboran Sea to southeastern Italy. Typical genetic breaks such as the Almeria-Oran Front or the Siculo-Tunisian Strait do not appear to be influential on the studied loci in H. margalefi sensu stricto. Instead, population structuring appears more complex and subtle than usually found for model species with a pelagic dispersal phase. The remaining four cryptic taxa are all found in the eastern basin, but incomplete lineage sorting is suspected and speciation might still be in process. Present-day population structure of the different H. margalefi cryptic species appears to result from past vicariance events started in the Oligo-Miocene and maintained by present-day coastal topography, water circulation and habitat fragmentation.


Monitoring contrasting land management in the savanna landscapes of northern Australia.
We compared measures of ecosystem state across six adjacent land-tenure groups in the intact tropical savanna landscapes of northern Australia. Tenure groups include two managed by Aboriginal owners, two national parks, a cluster of pastoral leases, and a military training area. This information is of relevance to the debate about the role of indigenous lands in the Australian conservation estate. The timing and frequency of fire was determined by satellite imagery; the biomass and composition of the herb-layer and the abundance of large feral herbivores by field surveys; and weediness by analysis of a Herbarium database. European tenures varied greatly in fire frequencies but were consistently burnt earlier in the dry season than the two Aboriginal tenures, the latter having intermediate fire frequencies. Weeds were more frequent in the European tenures, whilst feral animals were most abundant in the Aboriginal tenures. This variation strongly implies a signature of current management and/or recent environmental history. We identify indices suitable for monitoring of management outcomes in an extensive and sparsely populated landscape. Aboriginal land offers a unique opportunity for the conservation of biodiversity through the maintenance of traditional fire regimes. However, without financial support, traditional practices may prove unsustainable both economically and because exotic weeds and feral animals will alter fire regimes. An additional return on investment in Aboriginal land management is likely to be improved livelihoods and health outcomes for these disadvantaged communities.


PAK inactivation impairs social recognition in 3xTg-AD Mice without increasing brain deposition of tau and Aβ.
Defects in p21-activated kinase (PAK) are suspected to play a role in cognitive symptoms of Alzheimer's disease (AD). Dysfunction in PAK leads to cofilin activation, drebrin displacement from its actin-binding site, actin depolymerization/severing, and, ultimately, defects in spine dynamics and cognitive impairment in mice. To determine the role of PAK in AD, we first quantified PAK by immunoblotting in homogenates from the parietal neocortex of subjects with a clinical diagnosis of no cognitive impairment (n = 12), mild cognitive impairment (n = 12), or AD (n = 12). A loss of total PAK, detected in the cortex of AD patients (-39% versus controls), was correlated with cognitive impairment (r(2) = 0.148, p = 0.027) and deposition of total and phosphorylated tau (r(2) = 0.235 and r(2) = 0.206, respectively), but not with Aβ42 (r(2) = 0.056). Accordingly, we found a decrease of total PAK in the cortex of 12- and 20-month-old 3xTg-AD mice, an animal model of AD-like Aβ and tau neuropathologies. To determine whether PAK dysfunction aggravates AD phenotype, 3xTg-AD mice were crossed with dominant-negative PAK mice. PAK inactivation led to obliteration of social recognition in old 3xTg-AD mice, which was associated with a decrease in cortical drebrin (-25%), but without enhancement of Aβ/tau pathology or any clear electrophysiological signature. Overall, our data suggest that PAK decrease is a consequence of AD neuropathology and that therapeutic activation of PAK may exert symptomatic benefits on high brain function.


[Injury following sodium hypochlorite irrigation during endodontic treatment].
A 58-year-old woman was referred by her dentist to a maxillofacial surgeon because of a rapidly increasing facial swelling. The swelling developed after sodium hypochlorite irrigation during the endodontic treatment of tooth 25. A mechanical heart valve, a heart rhythm disorder, and antithrombotic therapy were complicating medical conditions. Treatment consisted of 12 mg dexamethason administered once intravenously, augmentin administered thrice daily intravenously, and oral analgetics. Damage following sodium hypochlorite irrigation during endodontic treatment is a rare disorder which is associated with a severe reaction in the surrounding tissue. Damage can be permanent.


Comparative in vitro activity of cefepime against strains isolated from intensive care and haematology patients.
The aim of the study was to evaluate cefepime, a "fourth generation" cephalosporin, for its in vitro activity in comparison with 5 other broad spectrum antibiotics against clinical isolates from ICU and haematology patients. The strains were isolated from blood (8%), lower respiratory tract (41%), upper respiratory tract (14%), urine (20%) and other sites (17%). They were divided into: 28 non-inducible Enterobacteriaceae, 35 inducible Enterobacteriaceae, 20 non-fermenters, 10 S. aureus and 10 Streptococcus spp. The MIC-values were determined by E-test. Overall, the rank order of susceptibility was cefepime (93%), imipenem (90%), piperacillin-tazobactam (81%), ciprofloxacin (79%), ceftriaxone (75%) and ceftazidime (74%). Only cefepime was able to inhibit all inducible Enterobacteriaceae. Against Pseudomonas aeruginosa, cefepime had the same activity as ceftazidime. With the exception of ceftazidime (65%), all beta lactams demonstrated good activity against Gram positive cocci. The excellent activity against most Gram negative and Gram positive pathogens suggests that cefepime may be useful in the treatment of serious infections in the described patient population.


A simple soft lithographic route to fabrication of poly(ethylene glycol) microstructures for protein and cell patterning.
We present a simple, direct soft lithographic method to fabricate poly(ethylene glycol) (PEG) microstructures for protein and cell patterning. This lithographic method involves a molding process in which a uniform PEG film is molded with a patterned polydimethylsiloxane stamp by means of capillary force. The patterned surfaces created by this method provide excellent resistance towards non-specific protein and cell adsorption. The patterned substrates consist of two regions: the molded PEG surface that acts as a resistant layer and the exposed substrate surface that promotes protein or cell adsorption. A notable finding here is that the substrate surface can be directly exposed during the molding process due to the ability to control the wetting properties of the polymer on the stamp, which is a key factor to patterning proteins and cells.


Quantifying ataxia: ideal trajectory analysis--a technical note.
We describe a quantitative method to assess repeated stair stepping stability. In both the mediolateral (ML) and anterioposterior (AP) directions, the trajectory of the subject's center of mass (COM) was compared to an ideal sinusoid. The two identified sinusoids were unique in each direction but coupled. Two dimensionless numbers-the mediolateral instability index (IML) and AP instability index (IAP)-were calculated using the COM trajectory and ideal sinusoids for each subject with larger index values resulting from less stable performance. The COM trajectories of nine nonimpaired controls and six patients diagnosed with unilateral or bilateral vestibular labyrinth hypofunction were analyzed. The average IML and IAP values of labyrinth disorder patients were respectively 127% and 119% greater than those of controls (p<0.014 and 0.006, respectively), indicating that the ideal trajectory analysis distinguishes persons with labyrinth disorder from those without. The COM trajectories also identify movement inefficiencies attributable to vestibulopathy.


Unexpected Oligomerization of Small α-Dicarbonyls for Secondary Organic Aerosol and Brown Carbon Formation.
Large amounts of small α-dicarbonyls (glyoxal and methylglyoxal) are produced in the atmosphere from photochemical oxidation of biogenic isoprene and anthropogenic aromatics, but the fundamental mechanisms leading to secondary organic aerosol (SOA) and brown carbon (BrC) formation remain elusive. Methylglyoxal is commonly believed to be less reactive than glyoxal because of unreactive methyl substitution, and available laboratory measurements showed negligible aerosol growth from methylglyoxal. Herein, we present experimental results to demonstrate striking oligomerization of small α-dicarbonyls leading to SOA and BrC formation on sub-micrometer aerosols. Significantly more efficient growth and browning of aerosols occur upon exposure to methylglyoxal than glyoxal under atmospherically relevant concentrations and in the absence/presence of gas-phase ammonia and formaldehyde, and nonvolatile oligomers and light-absorbing nitrogen-heterocycles are identified as the dominant particle-phase products. The distinct aerosol growth and light absorption are attributed to carbenium ion-mediated nucleophilic addition, interfacial electric field-induced attraction, and synergetic oligomerization involving organic/inorganic species, leading to surface- or volume-limited reactions that are dependent on the reactivity and gaseous concentrations. Our findings resolve an outstanding discrepancy concerning the multiphase chemistry of small α-dicarbonyls and unravel a new avenue for SOA and BrC formation from atmospherically abundant, ubiquitous carbonyls and ammonia/ammonium sulfate.


Evaluation of functional respiratory parameters in AIDS patients assisted in the infectious diseases ambulatory care clinic of a tertiary care university hospital in Brazil.
Pulmonary function, in HIV infected patients, has been associated with reduction in pulmonary ventilation parameters. We conducted a prospective cross sectional study to evaluate the pulmonary function of AIDS patients cared for in the infectious diseases ambulatory care clinic. Maximal inspiratory (P(Imax)) and expiratory (P(Emax)) pressures and spirometry were assessed. Clinical, demographic, and laboratory data were also evaluated. P(Imax) and P(Emax) were assessed in 73 and spirometry in 54 subjects. The mean time of HIV infection (in years) was similar for men (8.5 ± 5.1 y) and women (10.4 ± 4.2 y, P = .13), and the mean time of use of antiretroviral therapy was 8.5 ± 4.1 years for women and 7.7 ± 4.1 years for men (P = .46). P(Imax) and P(Emax) values were normal in 35 (48%) and 48 (66%) subjects, respectively. The use of tenofovir and presence of cough, by the time the test was performed, were independently associated with P(Emax) below the predicted value. Elevated creatine kinase values were associated with prolonged antiretroviral usage (9.6 ± 4.1 y vs 7.4 ± 3.9 y, P = .05). FVC was reduced in 14 (26.4%) subjects and was independently associated with high and/or intermediate cardiovascular risk (P = .002), and those with reduced vesicular murmur in auscultation (P = .047). FEV(1) was significantly lower in subjects with prolonged time of smoking (P = .02) and high and/or intermediate cardiovascular risk (P = .003). Reduced FEV(1)/FVC was associated with smoking (P = .041). AIDS subjects, in our study, had reduced parameters of maximal respiratory pressures and spirometry. The frequent dysfunction of respiratory muscles might be due to the association of multiple factors and not a particular one; moreover, smoking was independently associated with abnormal airway function. Pulmonary function tests should be implemented as an essential part of the medical assistance to AIDS patients.


Comparison of Serum Amyloid A Measurements in Equine Synovial Fluid With Routine Diagnostic Methods to Detect Synovial Infection in a Clinical Environment.
Synovial fluid analysis is utilized to diagnose septic synovitis. However, not all cases are clearly and rapidly discernible with the diagnostic tools available in the laboratory. Serum amyloid A (SAA), an acute phase protein, has been shown to be elevated in synovial fluid from inflamed synovial structures. The goal of this study is to describe the correlation between two diagnostic tests measuring equine SAA levels in septic and non-septic synovial structures and to understand the correlation between an elevated SAA result and synovial sepsis. Prospective estimation of sensitivity (Se) and specificity (Sp) of two tests, handheld and ELISA, measuring SAA in synovial fluid was completed in 62 horses presented with injured synovial structures. The comparison was made to a reference diagnosis based on white cell count, percentage of neutrophils, intracellular bacteria and bacterial culture on synovial fluid. Handheld test levels were classified as: 4 lines visible-SAA level negative; 3 lines visible-SAA level mild; 2 lines visible-SAA level moderate; and 1 line visible-SAA level severe and compared to the numerical value obtained with ELISA test. The ELISA SAA test had an area under the curve of 0.88 (0.78-0.98). An ELISA cut-off of 23.95 μg/mL maximized Se and Sp. This cutoff gave a Se of 0.93 (0.66-1.00) and Sp of 0.77 (0.63-0.88). The handheld test was highly correlated with the ELISA SAA test (Spearman rank correlation 0.96) and at a cutoff of moderate or higher for positive results gave identical Se and Sp. Se and Sp of synovial fluid SAA are very reliable when clinical signs of synovitis are present for >6 h. This test, in conjunction with traditional methods, can assist practitioners to rapidly diagnose and expedite appropriate intervention of synovial sepsis.


Distribution of carboxyhaemoglobin concentrations in smokers and non-smokers.
Carboxyhaemoglobin concentrations were measured in 3487 subjects comprising 1255 non-smokers, 1933 cigarette smokers, 193 cigar smokers (39 primary, 154 secondary), and 106 pipe smokers (30 primary, 76 secondary). In cigarette smokers the mean carboxyhaemoglobin concentration was 4.78% of the total haemoglobin and 94.7% of smokers had a concentration greater than 1.7%. Primary cigar smokers had a much lower mean carboxyhaemoglobin concentration (0.93%), and only 10.3% had concentrations greater than 1.7%. Primary pipe smokers also had a low mean carboxyhaemoglobin concentration (1.36%) and none had a concentration above 1.7%. Secondary cigar smokers had a high mean concentration (6.80%) and 97.4% had values above 1.7%; the findings in secondary pipe smokers were similar--the mean concentration was 3.39%, 94.7% having values greater than 1.7%. The lower carboxyhaemoglobin concentrations in primary pipe and cigar smokers suggest that in general they do not inhale, and the raised concentrations in cigarette smokers who change to pipes or cigars suggest that they usually continue to inhale and to absorb large amounts of carbon monoxide and other constituents of tobacco smoke.


Association of interleukin-28B polymorphisms with platelet count and liver function recovery after liver transplant.
The present genome-wide association study investigated the relationship of interleukin 28B (IL-28B) genetic variants with HBV susceptibility and prognosis of HBV-infected patients. This study aims to examine the role of IL-28B polymorphisms on transplant etiologies and the liver function recovery in Chinese liver transplant recipients.A total of 231 liver transplant recipients were enrolled in the study. The transplant etiologies included progressive HBV hepatitis, HBV-related liver cirrhosis (LC), HBV-related hepatocellular carcinoma (HCC), and non-HBV-related disease. All recipients were in stable condition before transplantation. Three single nucleotide polymorphisms (SNPs) of IL-28B (rs12979860, rs12980275, rs8099917) of recipients were analyzed by high-resolution melting (HRM) curve analysis. Liver function, blood cell count, and coagulation function were regularly tested before and for next 5 years after transplantation.No significant association was found between IL-28B gene polymorphisms and transplant etiologies. Peripheral platelet count in the third and fourth days after transplantation were significantly higher in recipients carrying IL-28B rs12979860 T allele, or rs8099917 C allele (P < .016666667), while there were no significant differences between these variants and International Normalized Ratio (INR) levels. In addition, gamma-glutamyltransferase (GGT) levels in recipients with rs12980275 G allele were higher than those in the wide-type recipients before transplantation (P < .016666667, respectively); nevertheless, no influence of these variants on GGT recovery was observed after transplantation.Genetic variations of IL-28B might impact on liver function recovery by influencing peripheral platelet counts and reducing liver inflammation, but have weak association with transplant etiologies.


LncRNAs-circRNAs as Rising Epigenetic Binary Superstars in Regulating Lipid Metabolic Reprogramming of Cancers.
As one of novel hallmarks of cancer, lipid metabolic reprogramming has recently been becoming fascinating and widely studied. Lipid metabolic reprogramming in cancer is shown to support carcinogenesis, progression, distal metastasis, and chemotherapy resistance by generating ATP, biosynthesizing macromolecules, and maintaining appropriate redox status. Notably, increasing evidence confirms that lipid metabolic reprogramming is under the control of dysregulated non-coding RNAs in cancer, especially lncRNAs and circRNAs. This review highlights the present research findings on the aberrantly expressed lncRNAs and circRNAs involved in the lipid metabolic reprogramming of cancer. Emphasis is placed on their regulatory targets in lipid metabolic reprogramming and associated mechanisms, including the clinical relevance in cancer through lipid metabolism modulation. Such insights will be pivotal in identifying new theranostic targets and treatment strategies for cancer patients afflicted with lipid metabolic reprogramming.


Effects of art therapy using color on purpose in life in patients with stroke and their caregivers.
Patients with stroke suffer from physical disabilities, followed by mental instability. Their caregivers also suffer from mental instability. The present study attempted to address the degree and the change of the level of Purpose in Life (PIL) in patients with stroke and caregivers by applying art therapy using colors. Twenty-eight stroke patients with a good functional recovery or a moderate disability and their 28 caregivers were selected and evaluated. The period of the study between the stroke and color therapy was more than 6 months. Patients and caregivers were divided into the color therapy (28) and control groups (28). A questionnaire, which measures the level of PIL was conducted separately for patients and caregivers prior to the first session of color therapy (2 hours per week, total 16 sessions). The final examination was performed 5 months after the last color therapy session. There was significant difference between before and after color therapy when the level of PIL was measured both in patients and caregivers (p<0.01). These were the same between the color therapy group, compared with the control group (p<0.01). As color therapy progressed to the late phase, patients and caregivers applied increasing number of colors and color intensity. These results prove that color therapy will improve PIL of the patients with post-stroke disability and caregivers. Furthermore, color therapy would be a useful adjuvant for improving the quality of life of the patients with stroke and their caregivers.


Opposite changes of pituitary and ovarian receptors for LHRH in ageing rats: further evidence for a direct neural control of ovarian LHRH receptor activity.
Hypophyseal and ovarian receptors for the neurohormone LHRH (LHRH-R) have been measured in young (3-4 months), middle-aged (8-11 months), constant estrous (CE, 10-14 months) and pseudopregnant (PR, 16-18 months) rats in order to study whether changes in hypothalamic and/or peripheral LHRH-like peptide production might precede and/or accompany the onset of reproductive failure observed in aging rats. Furthermore, we have investigated whether the neural efferent system from the brain to the ovary is affected with aging. The pattern of pituitary LHRH-R modifications during the estrous cycle of middle-aged rats shows lower LHRH-R levels on the second day of diestrus, resulting in a shift of the maximal LHRH binding capacity in the morning of proestrus. On the other hand, when comparing pituitary LHRH-R of animals exhibiting constant vaginal cornification (CVC) or repetitive PP with young estrus rats, no significant difference could be observed. Young rats responded to electrical stimulation (ES) of the medial preoptic area with an acute elevation of LHRH-R while ES performed in CVC, or PP animals resulted in a significant increase of hypophyseal LHRH binding capacity similar to the one observed in young estrous controls, indicating an impairment in the neural signals impinging in the pulse generating system, in old rats, and not an intrinsic defect of the LHRH-R per se. Ovarian LHRH-R concentration is higher in middle-aged cycling animals on the day of vaginal proestrus, compared to levels measured in young animals at any phase of the estrous cycle. Similarly, CE rats displaying CVC as well as PP animals show significantly higher numbers of LHRH-R with no change in affinity, than young estrus rats. CVC and PP rats receiving unilaterally an intraovarian injection of the potent LHRH antagonist, Ac-D-Cl-Phe1,2, D-Trp3, D-Phe6, D-Ala10, LHRH, showed an acute drop of LHRH-R measured within the treated ovary with no significant changes taking place in the vehicle-treated contralateral gland, suggesting that changes of endogenous ovarian LHRH-like peptide might participate in the mechanism(s) responsible for LHRH receptor increase observed in aging rats. In order to investigate the participation of a direct neural efferent pathway in ovarian LHRH-R regulation, young and old rats were subjected to spinal cord transection (above T10-T11). Bilateral transection of the spinal cord in young animals in the morning of proestrous markedly increased ovarian LHRH-R concentration in the afternoon (17.00 h) of the same day.(ABSTRACT TRUNCATED AT 400 WORDS)


[Evidence-based health care--critical inquiries from the medical ethics background].
Evidence-based medicine already plays an important role in clinical practice and in allocation decisions in health care. It is therefore of particular importance to acknowledge the ethical as well as the conceptual, methodological, epistemological and practical objections that have been articulated. A comprehensive medico-ethical evaluation of many of these caveats can be undertaken only in the context of a concrete health care system. The EU-sponsored EVIBASE project deals with these very issues. However, it is up to evidence-based medicine to prove it can comply with its own standards and counter the skeptical or critical arguments.


Clinical behavior of recurrent hormone receptor-positive breast cancer by adjuvant endocrine therapy within the Breast International Group 1-98 clinical trial.
Endocrine therapy resistance is a major cause of distant recurrence (DR) in hormone receptor-positive breast cancer. This study evaluated differences in survival after DR in patients treated with different adjuvant endocrine therapy regimens in the Breast International Group (BIG) 1-98 trial. BIG 1-98 compared 5 years of adjuvant treatment among 4 arms: tamoxifen (T), letrozole (L), tamoxifen followed by letrozole (TL), and letrozole followed by tamoxifen (LT). After a median follow-up of 8.1 years, 911 of 8010 patients (T, 302; L, 285; TL, 170; and LT, 154) had DR as the site of first recurrence. Univariate and multivariate Cox analyses were performed to determine features associated with post-DR survival. The median follow-up time after DR was 59 months (interquartile range, 29-88 months). Among all patients with DR, 38.1% were 65 years old or older at enrollment, 61.9% had tumors larger than 2 cm, and 69.7% were node positive. Neoadjuvant or adjuvant chemotherapy was administered to 35.6% of the patients. There was no difference in post-DR survival by treatment arm (median survival, 20.8 months for T, 17.9 months for L, 17.3 months for TL, and 20.8 months for LT; P = .21). In multivariate analysis, older patients (hazard ratio [HR], 1.35; 95% confidence interval [CI], 1.15-1.59) and patients with tumors larger than 2 cm (HR, 1.19; 95% CI, 1.00-1.41), 4 or more positive nodes (HR, 1.31; 95% CI, 1.05-1.64), progesterone receptor (PR)-negative tumors (HR, 1.25; 95% CI, 1.02-1.52), or shorter disease-free survival (DFS) had significantly worse post-DR survival. Treatment with adjuvant T, L, or their sequences was not associated with differences in survival after DR. Significant differences in survival were observed by age, primary tumor size, nodal and PR status, and DFS, and this suggests that traditional baseline high-risk features remain prognostic in the metastatic setting.


Complete endoscopic sphincterotomy with vs. without large-balloon dilation for the removal of large bile duct stones: randomized multicenter study.
Background and study aims Endoscopic sphincterotomy plus large-balloon dilation (ES-LBD) has been reported as an alternative to endoscopic sphincterotomy for the removal of bile duct stones. This multicenter study compared complete endoscopic sphincterotomy with vs. without large-balloon dilation for the removal of large bile duct stones. This is the first randomized multicenter study to evaluate these procedures in patients with exclusively large common bile duct (CBD) stones. Methods Between 2010 and 2015, 150 patients with one or more common bile duct stones ≥ 13 mm were randomized to two groups: 73 without balloon dilation (conventional group), 77 with balloon dilation (ES-LBD group). Mechanical lithotripsy was subsequently performed only if the stones were too large for removal through the papilla. Endoscopic sphincterotomy was complete in both groups. Patients could switch to ES-LBD if the conventional procedure failed. Results There was no between-group difference in number and size of stones. CBD stone clearance was achieved in 74.0 % of patients in the conventional group and 96.1 % of patients in the ES-LBD group (P < 0.001). Mechanical lithotripsy was needed significantly more often in the conventional group (35.6 % vs. 3.9 %; P < 0.001). There was no difference in terms of morbidity (9.3 % in the conventional group vs. 8.1 % in the ES-LBD group; P = 0.82). The cost and procedure time were not significantly different between the groups overall, but became significantly higher for patients in the conventional group who underwent mechanical lithotripsy. The conventional procedure failed in 19 patients, 15 of whom underwent a rescue ES-LBD procedure that successfully cleared all stones. Conclusions Complete endoscopic sphincterotomy with large-balloon dilation for the removal of large CBD stones has similar safety but superior efficiency to conventional treatment, and should be considered as the first-line step in the treatment of large bile duct stones and in rescue treatment.Trial registered at ClinicalTrials.gov (NCT02592811).


Low colonization rates of Clostridium difficile among patients and healthcare workers at Örebro University Hospital in Sweden.
The aim of this study was to investigate the rate of asymptomatic colonization rate of Clostridium difficile among both healthcare workers (HCWs) and patients in a hospital ward in Sweden. In a prospective observational study, asymptomatic HCWs (n = 22) (22/60; 37%) attending patients in an infectious disease ward in Sweden participated and were screened once for C. difficile. At the same time, 58 consecutive patients (58/227; 26%) admitted to the same ward were screened for C. difficile, first at admission and thereafter two times weekly. Fecal samples were obtained by rectal swabs and cultured anaerobically using both cycloserine-cefoxitin-fructose agar and enrichment (Cooked Meat broth). All samples were also tested by loop-mediated isothermal amplification and isolates were tested for the presence of toxin A or B by enzyme immunoassay. None of the analyzed fecal samples from HCWs contained C. difficile. Among the patients during a 2-month observational period, three of the 58 patients (5.2%) were culture positive regarding C. difficile on admission and one additional patient became asymptomatically colonized with C. difficile during the hospital stay. Thus, the colonization rates were 0% (0/22) (95% confidence interval (CI): 0-15.4%) among HCWs and 5.2% (3/58) (95% CI: 1.1-14.4%) among patients at admission. As the HCWs were screened only once, we have not studied transient colonization. In conclusion, with observed low colonization rates, we find no support that HCWs would be an important source for C. difficile transmission.


Nasopharyngeal Carriage Rate and Serotypes of Streptococcus pneumoniae and Antimicrobial Susceptibility in Healthy Korean Children Younger than 5 Years Old: Focus on Influence of Pneumococcal Conjugate Vaccination.
Even after pneumococcal vaccination introduction, Streptococcus pneumoniae (pneumoccocus) is still an important cause of respiratory and invasive severe infection. Pneumococcus is resided in nasal mucosa and local or systemic infection begins with the nasal mucosa damage. We studied the indirect effect of pneumococcal conjugate vaccine (PCV) on pneumococcal nasopharyngeal carriage rates, serotypes and antimicrobial susceptibility between vaccinate and non-vaccinated children. From January 2010 to October 2010, 379 healthy children under 5 years old from three university hospitals were recruited. Fully vaccinated children over 3 time doses of PCV and children with no vaccination history of PCV were enrolled, and nasopharyngeal aspirations were obtained from these children. Serotypes using multibead serotyping assay with multiplex PCR and antimicrobial susceptibility was analyzed. Antimicrobial susceptibilities were determined by the CLIS guideline. Two hundred seventy six children were received pneumococcal vaccination while 103 were not. 137 pneumococci were isolated from nasopharyngeal aspiration specimens. Nasal carriage rate was significantly low in vaccinated group (P-value; 0.001). Nasopharyngeal carriage rate was 28.6% (79/276) in vaccinate group and 56.3% (58/103) in non-vaccinated group. Among those vaccinated group, 13.0% (36/276) of the serotypes were vaccine or vaccine related type with the most common type 19F. In contrast, 31.1% (32/103) of the serotypes in non vaccinated group were vaccine or vaccine related type with the most common type 6A. The resistant rate of penicillin was 90.5%. For antimicrobial susceptibility, amoxicillin and amoxicillin/clavulanate showed high susceptibility (73.0%), but 19F and 19A serotypes were all resistant against amoxicillin. High nasopharyngeal carriage rate in non vaccinated group corresponded to the result of past study. However, 19F and 19A still came up as problematic serotypes with a high carriage rate and antimicrobial resistance in both vaccinated and non vaccinated groups. Also, this study showed that the resistance rate of primary oral antimicrobial agents was increased in compared to past. For solving these problems, the selective antimicrobial use with establishment of high dose amoxicillin/clavulanate regimen and active PCV immunization should be needed. Furthermore, pneumococcal carriage and serotype study concerning with antimicrobial susceptibility should be conducted in the future in 10 or 13-valent PCV received children.


Annexin A1 Regulates NLRP3 Inflammasome Activation and Modifies Lipid Release Profile in Isolated Peritoneal Macrophages.
Annexin A1 (AnxA1) is a potent anti-inflammatory protein that downregulates proinflammatory cytokine release. This study evaluated the role of AnxA1 in the regulation of NLRP3 inflammasome activation and lipid release by starch-elicited murine peritoneal macrophages. C57bl/6 wild-type (WT) and AnxA1-null (AnxA1-/-) mice received an intraperitoneal injection of 1.5% starch solution for macrophage recruitment. NLRP3 was activated by priming cells with lipopolysaccharide for 3 h, followed by nigericin (1 h) or ATP (30 min) incubation. As expected, nigericin and ATP administration decreased elicited peritoneal macrophage viability and induced IL-1β release, more pronounced in the AnxA1-/- cells than in the control peritoneal macrophages. In addition, nigericin-activated AnxA1-/- macrophages showed increased levels of NLRP3, while points of co-localization of the AnxA1 protein and NLRP3 inflammasome were detected in WT cells, as demonstrated by ultrastructural analysis. The lipidomic analysis showed a pronounced release of prostaglandins in nigericin-stimulated WT peritoneal macrophages, while ceramides were detected in AnxA1-/- cell supernatants. Different eicosanoid profiles were detected for both genotypes, and our results suggest that endogenous AnxA1 regulates the NLRP3-derived IL-1β and lipid mediator release in macrophages.


Sphingosine kinase type 1 inhibition reveals rapid turnover of circulating sphingosine 1-phosphate.
S1P (sphingosine 1-phosphate) is a signalling molecule involved in a host of cellular and physiological functions, most notably cell survival and migration. S1P, which signals via a set of five G-protein-coupled receptors (S1P1-S1P5), is formed by the action of two SphKs (sphingosine kinases) from Sph (sphingosine). Interfering RNA strategies and SphK1 (sphingosine kinase type 1)-null (Sphk1-/-) mouse studies implicate SphK1 in multiple signalling cascades, yet there is a paucity of potent and selective SphK1 inhibitors necessary to evaluate the effects of rapid onset inhibition of this enzyme. We have identified a set of submicromolar amidine-based SphK1 inhibitors and report using a pair of these compounds to probe the cellular and physiological functions of SphK1. In so doing, we demonstrate that our inhibitors effectively lower S1P levels in cell-based assays, but we have been unable to correlate SphK1 inhibition with changes in cell survival. However, SphK1 inhibition did diminish EGF (epidermal growth factor)-driven increases in S1P levels and Akt (also known as protein kinase B)/ERK (extracellular-signal-regulated kinase) phosphorylation. Finally, administration of the SphK1 inhibitor to wild-type, but not Sphk1-/-, mice resulted in a rapid decrease in blood S1P levels indicating that circulating S1P is rapidly turned over.


[Treatment of chronic snoring with CO2 laser].
Laser-assisted uvulopalatoplasty (LAUP) is an effective surgical procedure for treating habitual snoring. It is performed under local anesthesia as an outpatient procedure and consists of enlarging the oropharyngeal air space and restructuring the uvula, velum, and pharyngeal pillars. We used LAUP in 112 patients with upper airway resistance syndrome (UARS) (Lipman grade II) over the last three years and achieve a care rate of 87.5%.


In situ hybridization methods for the detection of somatostatin mRNA in tissue sections using antisense RNA probes.
In situ hybridization studies with [32P] and [3H] labelled antisense RNA probes were undertaken to determine optimal methods of tissue fixation, tissue sectioning, and conditions of hybridization, and to compare the relative merits of the two different radioactive labels. The distribution of somatostatin mRNA in neurons of rat brain using a labelled antisense somatostatin RNA probe was employed as a model for these studies. The highest degree of sensitivity for in situ hybridization was obtained using paraformaldehyde fixation and vibratome sectioning. Optimal autoradiographic localization of mRNA was obtained within 7 days using [32P] labelled probes. However, due to the high energy emittance of [32P], precise intracellular localization of hybridization sites was not possible. [3H] labelled RNA probes gave more precise cellular localization but required an average of 18-20 days autoradiographic exposure. The addition of the scintillator, PPO, decreased the exposure time for the localization of [3H] labelled probes to seven days. We also report a method for combined in situ hybridization and immunocytochemistry for the simultaneous localization of somatostatin in mRNA and peptide in individual neurons.


Increased susceptibility to beta-lactam antibiotics and decreased porin content caused by envB mutations of Salmonella typhimurium.
Isogenic derivatives carrying envB6, envB9, or envB+ alleles were obtained from a strain of Salmonella typhimurium that was partially resistant to mecillinam, a beta-lactam antibiotic specific for penicillin-binding protein 2 (PBP 2). Testing of the isogenic strains with several antibacterial agents demonstrated that envB mutations either increased resistance (mecillinam) or did not affect the response (imipemen) to beta-lactams that act primarily on PBP 2, while susceptibilities to beta-lactams that act on PBP 1B, PBP 3, or both were increased. Furthermore, the susceptibilities of envB strains to hydrophobic compounds such as rifampin, novobiocin, or chloramphenicol were not modified, even though their susceptibilities to deoxycholate and crystal violet were enhanced. Outer cell membranes of envB mutants presented a 50% reduction in protein content compared with that of the isogenic envB+ strains, and OmpF and OmpD porins were particularly affected by the reduction. No alteration in the amount or pattern of periplasmic proteins was noticed, and lipopolysaccharides from envB mutants appeared to be normal by sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis. By using derivatives that produced a plasmid-encoded beta-lactamase, it was demonstrated that envB cells are slightly less permeable to cephalothin than envB+ bacteria are. It is concluded that the high susceptibility of envB mutants to beta-lactams is due to the increased effectiveness of the antibiotics on PBP 1B, PBP 3, or both.


Optimal design and experimental validation of synchronous, asynchronous and flow-modulated, simulated moving-bed processes using a single-column setup.
The performance of asynchronous (Varicol) and flow-modulated (PowerFeed) simulated moving-bed processes, as well as their combination into a single hybrid scheme, are studied both experimentally and by numerical simulation. A recently developed single-column experimental setup is employed to demonstrate the feasibility of the various schemes, explore the effect of their major operating parameters, and illustrate the performance enhancements that are obtained when these schemes are properly optimized. The experimental feasibility and effectiveness of the various schemes are assessed by running and comparing optimized configurations for the linear separation of two nucleosides on a high-performance reversed-phase stationary phase.


Postmortem imaging of perimortem skeletal trauma.
Various imaging modalities, including conventional radiography, computed tomography, magnetic resonance, and surface scanning have been applied in the examination of skeletal injuries in the forensic context. Although still not a substitute for a full medico-legal autopsy or the examination of skeletal remains, imaging is now increasingly used as a complementary tool in the postmortem analysis of perimortem skeletal trauma. Facilitated by the progress in general computational capacity, multimodal imaging has been proposed for comprehensive forensic documentation. A major advantage of these imaging approaches is that stored digital or physical 3D models of skeletal injuries can be reviewed at any time by various experts as well as be presented in court as evidence to clarify potentially complex medical and forensic aspects of the case. Due to constant technical progress in imaging techniques and software, continuous education, training, and sharing of expertise among engineers, computer scientists, and forensic experts, including forensic pathologists, anthropologists, and radiologists needs to be warranted to maintain high-quality expertise in the detection and interpretation of traumatic injuries on postmortem imaging. The technical developments and ever-improving user-friendliness of 3D imaging and modeling techniques present an atttactive alternative to traditional forensic approaches, but as long as the techniques have not been sufficiently tested and validated for forensic trauma analysis, and best practice manuals for forensic practice are lacking for both the technical procedures and method selection, the use of imaging techniques needs to be reevaluated on a case-by-case basis. In addition, ethical, legal, and financial aspects of the use of imaging and 3D modeling for forensic purposes need to be well understood by all parties in legal proceedings.


Altered neurochemical coupling in the occipital cortex in migraine with visual aura.
Visual aura is present in about one-third of migraine patients and triggering by bright or flickering lights is frequently reported. Using migraine with visual aura patients, we investigated the neurochemical profile of the visual cortex using magnetic resonance spectroscopy. Specifically, glutamate/creatine and GABA/creatine ratios were quantified in the occipital cortex of female migraine patients. GABA levels in the occipital cortex of migraine patients were lower than that of controls. Glutamate levels in migraine patients, but not controls, correlated with the blood-oxygenation-level-dependent (BOLD) signal in the primary visual cortex during visual stimulation. Migraine with visual aura appears to disrupt the excitation-inhibition coupling in the occipital cortex.


Inhibitor selectivity of a new class of oseltamivir analogs against viral neuraminidase over human neuraminidase enzymes.
The viral neuraminidase enzyme is an established target for anti-influenza pharmaceuticals. However, viral neuraminidase inhibitors could have off-target effects due to interactions with native human neuraminidase enzymes. We report the activity of a series of known inhibitors of the influenza group-1 neuraminidase enzyme (N1 subtype) against recombinant forms of the human neuraminidase enzymes NEU3 and NEU4. These inhibitors were designed to take advantage of an additional enzyme pocket (known as the 150-cavity) near the catalytic site of certain viral neuraminidase subtypes (N1, N4 and N8). We find that these modified derivatives have minimal activity against the human enzymes, NEU3 and NEU4. Two compounds show moderate activity against NEU3, possibly due to alternative binding modes available to these structures. Our results reinforce that recognition of the glycerol side-chain is distinct between the viral and human NEU enzymes, and provide experimental support for improving the selectivity of viral neuraminidase inhibitors by exploiting the 150-cavity found in certain subtypes of viral neuraminidases.


ApoE polymorphism in a small Mediterranean island: relationships with plasma lipids, lipoproteins and LDL particle size.
Polymorphisms of apoE gene are able to modulate lipoprotein metabolism at different steps and to influence LDL-cholesterol (LDL-C) levels and also other lipoproteins features. Population studies documented large differences in the frequency of apoE alleles which could be even related to the prevalence of cardiovascular disease. In this study we evaluated the apoE genotypes and allele frequency in 576 subjects living in a small island in the Tyrrhenian Sea and the relative contribution of apoE polymorphism on plasma lipid and lipoprotein profile, including LDL particle size. We found a cumulative frequency of 0.073, 0.866 and 0.061 for epsilon2, epsilon3 and epsilon4 alleles respectively. Moreover epsilon3 subjects had only triglyceride levels significantly lower and LDL-C and lipoprotein (a) (Lp(a)) levels higher than epsilon2 carriers. LDL-particle size was significant smaller in epsilon2 subjects than both epsilon3 and epsilon4 carriers, but the difference disappeared when data were adjusted for triglycerides. In conclusion we have provided further evidence of a low prevalence of epsilon4 allele in a Mediterranean population which may represent a genetic protective factor of these populations. Environmental factors, such as diet, occurring in this area may have attenuated the influence of this gene on plasma lipoproteins.


Chip-based polyketide biosynthesis and functionalization.
We demonstrate construction and novel compound synthesis from a synthetic metabolic pathway consisting of a type III polyketide synthase (PKS) known as 1,3,6,8-tetrahydroxynaphthalene synthase (THNS) from Streptomyces coelicolor and soybean peroxidase (SBP) in a microfluidic platform. THNS immobilized to Ni-NTA agarose beads is prepacked into a microfluidic channel, while SBP is covalently attached to the walls of a second microfluidic channel precoated with a reactive poly(maleic anhydride) derivative. The result is a tandem, two-step biochip that enables the synthesis of novel polyketide derivatives. The first microchannel, consisting of THNS, results in the conversion of malonyl-CoA to flaviolin in yields up to 40% with a residence time of 6 min. This conversion is similar to that obtained in several-milliliter batch reactions after 2 h. Linking this microchannel to the SBP microchannel results in biflaviolin synthesis. During the course of this work, we discovered that the substrate specificity of THNS could be manipulated by simply changing the reaction pH. As a result, the starter acyl-CoA specificity can be broadened to yield a series of truncated pyrone products. When combined with variations in the ratio of acyl-CoA and malonyl-CoA (extender substrate) feed rates, high yields of the pyrone products could be achieved, which is further structurally diversified from self- and cross-coupling in the SBP microchannel. The ability to rapidly evaluate the effects of reaction conditions and synthetic multienzyme pathways on a microfludic platform provides a new paradigm for performing metabolic pathway engineering, namely, the reconstruction of pathways for use in new compound discovery.


Differential avian and human tuberculin skin testing in non-tuberculous mycobacterial infection.
To determine the sensitivity of differential avian and human delayed-type hypersensitivity skin testing in the diagnosis of non-tuberculous mycobacterial lymphadenitis. Retrospective review of all patients with culture proved non-tuberculous mycobacterial lymph node infections who also had differential avian and human skin testing performed over a 10 year period from 1986 to 1996. One hundred and twenty four patients had non-tuberculous mycobacteria isolated from lymph nodes over this period, 59 of whom had differential skin testing performed. The sensitivity of a response of >/= 10 mm to the avian precipitin was 58 of 59. No patient had both a negative human and avian Mantoux. The sensitivity of the human Mantoux alone for diagnosing non-tuberculous mycobacterial infection was 81% for a response of >/= 5 mm and 66% for >/= 10 mm. Ten patients had a 0 human response. Fifty five of the 59 patients had an avian response at least 2 mm greater than the human response. The avian Mantoux is a very sensitive method of diagnosing non-tuberculous mycobacterial infection in children. The human Mantoux is not sensitive enough to be used alone as a surrogate to diagnose non-tuberculous mycobacterial infection.


Impaired protein aggregate handling and clearance underlie the pathogenesis of p97/VCP-associated disease.
Mutations in p97/VCP cause the multisystem disease inclusion body myopathy, Paget disease of the bone and frontotemporal dementia (IBMPFD). p97/VCP is a member of the AAA+ (ATPase associated with a variety of activities) protein family and has been implicated in multiple cellular processes. One pathologic feature in IBMPFD is ubiquitinated inclusions, suggesting that mutations in p97/VCP may affect protein degradation. The present study shows that IBMPFD mutant expression increases ubiquitinated proteins and susceptibility to proteasome inhibition. Co-expression of an aggregate prone protein such as expanded polyglutamine in IBMPFD mutant cells results in an increase in aggregated protein that localizes to small inclusions instead of a single perinuclear aggresome. These small inclusions fail to co-localize with autophagic machinery. IBMPFD mutants avidly bind to these small inclusions and may not allow them to traffic to an aggresome. This is rescued by HDAC6, a p97/VCP-binding protein that facilitates the autophagic degradation of protein aggregates. Expression of HDAC6 improves aggresome formation and protects IBMPFD mutant cells from polyglutamine-induced cell death. Our study emphasizes the importance of protein aggregate trafficking to inclusion bodies in degenerative diseases and the therapeutic benefit of inclusion body formation.


Prion Strain-Specific Structure and Pathology: A View from the Perspective of Glycobiology.
Prion diseases display multiple disease phenotypes characterized by diverse clinical symptoms, different brain regions affected by the disease, distinct cell tropism and diverse PrPSc deposition patterns. The diversity of disease phenotypes within the same host is attributed to the ability of PrPC to acquire multiple, alternative, conformationally distinct, self-replicating PrPSc states referred to as prion strains or subtypes. Structural diversity of PrPSc strains has been well documented, yet the question of how different PrPSc structures elicit multiple disease phenotypes remains poorly understood. The current article reviews emerging evidence suggesting that carbohydrates in the form of sialylated N-linked glycans, which are a constitutive part of PrPSc, are important players in defining strain-specific structures and disease phenotypes. This article introduces a new hypothesis, according to which individual strain-specific PrPSc structures govern selection of PrPC sialoglycoforms that form strain-specific patterns of carbohydrate epitopes on PrPSc surface and contribute to defining the disease phenotype and outcomes.


A dichroic surface-plasmon-polariton splitter based on an asymmetric T-shape nanoslit.
An asymmetric T-shape nanoslit in a metal film is proposed to act as an efficient dichroic surface-plasmon-polariton (SPP) splitter, which is composed of a single nanoslit in immediate contacting with two nanogrooves with different widths. Simulations show that, due to the interferences of SPPs in the upper part of the asymmetric T-shape nanoslit, the generated SPPs propagating to the left and right directions on the front metal surface can be manipulated nearly independently by altering the right and left groove widths, respectively. Based on such effects, a dichroic SPP splitter is demonstrated and the splitting wavelengths can easily be adjusted. High splitting ratios of 31:1 and 1:12 at splitting wavelengths of 680 nm and 884 nm are numerically presented with a device's lateral dimension of only 1200 nm. Further experimental results match the simulations well.


Epinephrine increases plasma immunoreactive atrial natriuretic hormone levels in humans.
Six normal human subjects each underwent sequential 80-min infusions of saline and epinephrine (EPI) at 0.55 and 2.75 micrograms X min-1 X m-2 to determine the role of EPI in the control of atrial natriuretic hormone (ANH) in humans. Plasma immunoreactive-ANH (IR-ANH) levels nearly doubled in response to the infusion of EPI at 0.55 microgram X min-1 X m-2 (P less than 0.05) and then plateaued; heart rate accelerated significantly (P less than 0.01) with increasing plasma EPI levels, whereas systolic blood pressure increased only with higher plasma EPI levels (P less than 0.05). To determine whether beta-adrenergic mechanisms mediate the EPI-induced increase in IR-ANH, six additional subjects each received infusions on two separate days of saline for 240 min and the beta-adrenergic antagonist propranolol followed by propranolol plus EPI at 2.75 micrograms X min-1 X min-2 each for 80 min. Neither saline nor propranolol plus EPI caused a significant increase in plasma IR-ANH. We conclude that EPI increases plasma IR-ANH through beta-adrenergic mechanisms in humans. beta-Adrenergic-mediated increases in plasma IR-ANH levels appear to be unrelated to changes in the heart rate.


Dissection of the role of the stable signal peptide of the arenavirus envelope glycoprotein in membrane fusion.
The arenavirus envelope glycoprotein (GPC) retains a stable signal peptide (SSP) as an essential subunit in the mature complex. The 58-amino-acid residue SSP comprises two membrane-spanning hydrophobic regions separated by a short ectodomain loop that interacts with the G2 fusion subunit to promote pH-dependent membrane fusion. Small-molecule compounds that target this unique SSP-G2 interaction prevent arenavirus entry and infection. The interaction between SSP and G2 is sensitive to the phylogenetic distance between New World (Junín) and Old World (Lassa) arenaviruses. For example, heterotypic GPC complexes are unable to support virion entry. In this report, we demonstrate that the hybrid GPC complexes are properly assembled, proteolytically cleaved, and transported to the cell surface but are specifically defective in their membrane fusion activity. Chimeric SSP constructs reveal that this incompatibility is localized to the first transmembrane segment of SSP (TM1). Genetic changes in TM1 also affect sensitivity to small-molecule fusion inhibitors, generating resistance in some cases and inhibitor dependence in others. Our studies suggest that interactions of SSP TM1 with the transmembrane domain of G2 may be important for GPC-mediated membrane fusion and its inhibition.


Doxycycline or moxifloxacin for the management of community-acquired pneumonia in the UK?
Macrolide monotherapy is currently recommended as first-line treatment of mild-to-moderate community-acquired pneumonia (CAP) in penicillin-intolerant/allergic individuals in the UK. However, resistance rates among the commonest cause, Streptococcus pneumoniae, now exceed 10% in the UK and a review of alternative agents is therefore timely. This review considers the relative merits of two agents, doxycycline and moxifloxacin, which are candidates to replace macrolides for second-line therapy of non-severe CAP.


"Mosaic" medtronic bioprosthetic valve replacement clinical results and hemodynamical performance.
We report the mid-term results of a prospective trial of a new bioprosthetic valve. The Mosaic bioprosthesis consists of porcine aortic valve that has been cross linked ed in glutaraldehyde solution under zero-pressure fixation and treated with alpha amino oleic acid to reduce the potential for calcification. Mosaic bioprosthetic valve replacement was performed in 67 consecutive patients between January 1995 and August 1998. There were 37 patients having aortic valve replacement (AVR) and 30 having mitral valve replacement (MVR) who entered this study. The patients age ranged 56 to 86 years (mean 74.9); 38 were female and 29 were male; 44 were in NYHA grade 3 and 21 were NYHA grade 4. All mitral valve replacements were performed with total preservation of subvalvular apparatus. Echocardiographic assessment of valve and LV function were performed on 7th day, 6 months 1,2 and 3 years. There was no hospital mortality. 3 year survival was 85.9+/-5.9% for AVR and 100% for MVR. Freedom from antithromboembolic related haemorrhage has been 96.7% for MVR and 91.9% for AVR. Freedom from the transient neurological event was 96.7+/-3.3% for MVR and 100% for AVR Freedom from structural valve failure, permanent thromboembolism, thrombosis or endocarditis has been 100% for both AVR and MVR. In AVR group left ventricle mass, left ventricle mass index significantly decreased, when cardiac index and effective orifice area increased significantly during study period. Transvalvular gradient did not change. In MVR group transvalvular gradient, effective orifice area and cardiac index did not change. The valve was user friendly. The early results are very satisfactory. Echocardiography measurements after aortic valve replacement are showing very marked late postoperative remodelling of left ventricle. After mitral valve replacement there were exceptionally low transvalvular gradients, no left ventricle outflow tract obstruction.


Monitoring of pregnancies at risk for Fanconi's anemia by chorionic villi sampling.
Two pregnancies at risk for Fanconi's anemia have been monitored by a cytogenetic method in the first trimester of gestation. The rate of chromosome breakage was evaluated in spontaneous mitoses from a direct preparation of trophoblasts in one case and from mitoses obtained from standard cultures and from mitoses treated with diepoxybutane in both cases. Cytogenetic studies were carried out also at 16 weeks from amniotic fluid cells in one case, and in fibroblasts sampled from the aborted fetus in the other. In all the experimental conditions the mean frequency of breaks/cell was in the range of controls, suggesting that the fetuses were unaffected by Fanconi's anemia. In one case the results have been confirmed by chromosome analysis at birth.


A ketogenic diet for reducing obesity and maintaining capacity for physical activity: hype or hope?
This review will evaluate the recent scientific literature on the efficacy of the ketogenic diet for enhancing weight loss and maintaining capacity for physical activity. With the increasing obesity epidemic, the 21st century has witnessed the emergence of various diet programs aimed at promoting weight loss and enhancing physical performance. The ketogenic diet, a diet high in fat and low in carbohydrates, has been at the forefront. Recent literature has documented significant weight-loss following ketogenic diet interventions. This has been associated with a decrease in systemic inflammation, reduced insulin resistance, and improved lipid profile. Given the importance of physical activity to sustaining a healthy weight, the impact of a ketogenic diet on physical performance has been investigated. The majority of the current literature supports a positive influence of the ketogenic diet on physical activity owing to its ability to maintain performance of both endurance and resistance activities. We highlight the most important recent advances that have been made on the ketogenic diet in relation to obesity and capacity for physical activity. The ketogenic diet is a potentially promising diet for reducing obesity while maintaining capacity for physical activity. However, questions surrounding the sustainability of this diet for the long-term remain.


Experience and Outcomes of Primary Percutaneous Coronary Intervention for Patients with ST-Segment Elevation Myocardial Infarction of Tertiary Care Center in Bosnia and Herzegovina.
Primary percutaneous coronary intervention (PCI) is an emergent percutaneous catheter intervention in the setting of ST-segment elevations myocardial infarction (STEMI), without previous fibrinolytic treatment. To evaluate the feasibility and outcomes of primary percutaneous coronary interventions for STEMI in regional tertiary care cardiac centre in Bosnia and Herzegovina. Between January 2014 and December 2016, consecutive 549 STEMI patients who underwent primary PCI were prospectively enrolled in a primary PCI registry. The most of coronary angiography procedures were performed using the radial artery route. Patient demographics, risk factors, procedural characteristics, time variables and in-hospital events were assessed. On admission, 297 (64.7%) of the patients were current smokers, 234 (42.6%) were hypertensive, 172 (31.3%) were diabetics, and 68 (12.3%) had cardiogenic shock. The mean duration of time from symptom onset to hospital arrival 193±118.2 minutes, and the mean door-to-balloon time was 37±11.3 minutes and median total ischemic time was 265(60-897) minutes. Infarct-related artery (IRA) was the left anterior descending artery in 47.1%, multivessel disease was present in 49.7%. Primary PCI involved balloon dilatation (2.7%) and stent implantation (97.3%). The incidence of postprocedural angiographic no-reflow was 6.7%. All-cause mortality occurred in 17 patients (3.1%). This study has shown feasibility and efficiency in performing of primary PCI with good outcomes in the first regional interventional center in Bosnia and Herzegovina. Experiences and results of our hospital can be very useful in creating primary PCI networks in our countries and developing countries as well.


Mental disorders in Bangladesh: a systematic review.
Mental disorders constitute a major public health problem globally with higher burden in low and middle-income countries. In Bangladesh, systematically-collected data on mental disorders are scarce and this leaves the extent of the problem not so well defined. We reviewed the literature on mental health disorders in Bangladesh to summarize the available data and identify evidence gaps. We identified relevant literature on mental disorders within Bangladesh published between 1975 and October, 2013 through a systematic and comprehensive search. Relevant information from the selected articles was extracted and presented in tables. We identified 32 articles which met our pre-defined eligibility criteria. The reported prevalence of mental disorders varied from 6.5 to 31.0% among adults and from 13.4 to 22.9% among children. Some awareness regarding mental health disorders exists at community level. There is a negative attitude towards treatment of those affected and treatment is not a priority in health care delivery. Mental health services are concentrated around tertiary care hospitals in big cities and absent in primary care. The burden of mental disorders is high in Bangladesh, yet a largely unrecognized and under-researched area. To improve the mental health services in Bangladesh, further well-designed epidemiological and clinical research are needed.


Coordination of glioblastoma cell motility by PKCι.
Glioblastoma is one of the deadliest forms of cancer, in part because of its highly invasive nature. The tumor suppressor PTEN is frequently mutated in glioblastoma and is known to contribute to the invasive phenotype. However the downstream events that promote invasion are not fully understood. PTEN loss leads to activation of the atypical protein kinase C, PKCι. We have previously shown that PKCι is required for glioblastoma cell invasion, primarily by enhancing cell motility. Here we have used time-lapse videomicroscopy to more precisely define the role of PKCι in glioblastoma. Glioblastoma cells in which PKCι was either depleted by shRNA or inhibited pharmacologically were unable to coordinate the formation of a single leading edge lamellipod. Instead, some cells generated multiple small, short-lived protrusions while others generated a diffuse leading edge that formed around the entire circumference of the cell. Confocal microscopy showed that this behavior was associated with altered behavior of the cytoskeletal protein Lgl, which is known to be inactivated by PKCι phosphorylation. Lgl in control cells localized to the lamellipod leading edge and did not associate with its binding partner non-muscle myosin II, consistent with it being in an inactive state. In PKCι-depleted cells, Lgl was concentrated at multiple sites at the periphery of the cell and remained in association with non-muscle myosin II. Videomicroscopy also identified a novel role for PKCι in the cell cycle. Cells in which PKCι was either depleted by shRNA or inhibited pharmacologically entered mitosis normally, but showed marked delays in completing mitosis. PKCι promotes glioblastoma motility by coordinating the formation of a single leading edge lamellipod and has a role in remodeling the cytoskeleton at the lamellipod leading edge, promoting the dissociation of Lgl from non-muscle myosin II. In addition PKCι is required for the transition of glioblastoma cells through mitosis. PKCι therefore has a role in both glioblastoma invasion and proliferation, two key aspects in the malignant nature of this disease.


The recovery vision as a service improvement guide for community mental health center providers.
Many mental health consumers are calling for Community Mental Health Center services to focus on creating opportunities for recovery. First person accounts of recovery portray a process which entails developing hope, taking personal responsibility for health, and building a broad sense of self that is not illness-dominated. Community Mental Health Centers can facilitate recovery by enhancing their clinicians' abilities to (1) communicate hope, (2) teach the skills and knowledge consumers need to effectively manage their health concerns, and (3) support consumers' efforts to get on with life beyond illness.


Laparoscopically assisted vaginal hysterectomy (LAVH) versus total abdominal hysterectomy (TAH) in endometrial carcinoma: prospective cohort study.
To determine the feasibility and safety of laparoscopically assisted vaginal hysterectomy in the treatment of presumed stage I endometrial cancer. This was a prospective cohort study without randomization of 182 consecutive patients who underwent surgery for early endometrial cancer or atypical hyperplasia at the West Kent Gynaecological Oncology Centre, UK. Seventy-four had laparoscopically assisted vaginal hysterectomy and bilateral salpingo-oophorectomy (BSO), and 108 had a total abdominal hysterectomy and bilateral salpingo-oophorectomy. Lymphadenectomy was performed in 153 patients, and lymph node sampling was performed in 2 patients. Twenty-seven patients with serous papillary endometrial cancer in addition had an omentectomy. The groups were compared for epidemiological and clinical characteristics, surgical outcomes, hospital stay, lymph node harvest, and intraoperative and postoperative complications. The patients in the laparoscopy group had less blood loss, similar number of lymph nodes removed, less need for analgesia, and shorter hospital stay but longer operative time than those treated by laparotomy. In our study, we had 4 conversions (5.4%) from laparoscopy to laparotomy. Twenty-eight (41%) patients who had laparoscopic surgery were obese (body mass index [BMI] >30 kg/m2). Postoperative complications were more common in the laparotomy group (34%) than in the laparoscopy group (6%). No major complications occurred in the laparoscopy group. Wound infection was the most common complication in laparotomy patients, and this invariably happened to obese patients (BMI >30 kg/m2). There were 6 readmissions, all from the laparotomy group. Laparoscopic surgery is a safe and reliable alternative to open surgery in the management of early endometrial cancer patients, with significantly reduced hospital stay and complications, especially in those patients with an elevated BMI.


[Aquaporin-4].
In human body, there are thirteen water channels but their expression patterns are tissue specific. Aquaporin-4 (AQP4) is a predominantly expressed water channel in the mammalian brain and an important drug target for treatment of cerebral edema, bipolar disorder, and mesial temporal lobe epilepsy. Recently it was reported that IgG of optic-spinal multiple sclerosis patients bound to AQP4. In order to reveal the function of AQP4, we determined the atomic structure of AQP4 by electron crystallography of double layered two-dimensional crystals. In double layered crystal, each single layered crystal contacts by a short 310 helix in the extracellular loop C. It would suggest that AQP4 shows the weak adhesive activity between adjoining membranes. This is correlated to immunogold labeling of AQP4 in glial lamellae localizing the protein areas where the membranes are separated but also all along junctional regions. Furthermore, from the freeze fracture replica labeling and the mutational experiment, the palmitoylation of N-terminal cysteine residues makes orthogonal array structure unstable on Chinese hamster Ovary (CHO) cell membrane. These findings suggest that there must be the complicated mechanism for control of water content relevant to AQP4 within the brain.


Optimal management of nasal congestion caused by allergic rhinitis in children: safety and efficacy of medical treatments.
Nasal congestion is such a frequent and multifactorial occurrence in young children that parents and medical caregivers often overlook the need for medical intervention. However, children with congestion can suffer quality-of-life detriments resulting from sleep disturbance, learning impairment, and fatigue. Congestion also impairs the normal nasal breathing that is physiologically important for the efficient cleaning and conditioning of inspired air. Further, the most common cause of congestion, allergic rhinitis, is considered a potential risk factor for asthma. Published guidelines on the treatment of allergic rhinitis agree that management strategies in children should follow the same principles as in adults, while recognizing the need for dosage adjustments and being aware of unique safety issues. Intranasal corticosteroids, with robust effects in reducing congestion and good tolerability, remain a treatment of choice. Despite lingering concerns about the potential for growth suppression with these drugs, clinical evidence suggests a very low risk at prescribed dosages, especially with compounds that have a low systemic bioavailability. Oral antihistamines are commonly cited as first-line options for allergic rhinitis, although their effect on nasal congestion is relatively modest. First-generation antihistamines should not be administered to children because of their sedative properties, which can worsen learning problems associated with allergic rhinitis. Second-generation oral antihistamines are preferred, although this class is not completely devoid of adverse effects. Other treatments, such as a nasal antihistamine, decongestants, and immunotherapy, present varying levels of safety and tolerability issues in children.


Development of minimum criteria for the initiation of antibiotics in residents of long-term-care facilities: results of a consensus conference.
Establishing a clinical diagnosis of infection in residents of long-term-care facilities (LTCFs) is difficult. As a result, deciding when to initiate antibiotics can be particularly challenging. This article describes the establishment of minimum criteria for the initiation of antibiotics in residents of LTCFs. Experts in this area were invited to participate in a consensus conference. Using a modified delphi approach, a questionnaire and selected relevant articles were sent to participants who were asked to rank individual signs and symptoms with respect to their relative importance. Using the results of the weighting by participants, a modification of the nominal group process was used to achieve consensus. Criteria for initiating antibiotics for skin and soft-tissue infections, respiratory infections, urinary infections, and fever where the focus of infection is unknown were developed.


Differential blood cells associated with parasitism in the wild puffer fish Lagocephalus laevigatus (Tetraodontiformes) of the Campeche Coast, southern Mexico.
In aquaculture conditions, severe parasitic infections cause negative impacts on fish health and economic losses. The parasite load has been associated with anemia, which reduces the number of erythrocytes in fish. Therefore, the evaluation of hematological parameters as a feasible tool for diagnosing and monitoring fish health allows us to determine the indirect effect of parasites on the health status of fish. Our aim was to evaluate changes in the blood cell parameters of Lagocephalus laevigatus associated with parasitism. A total of 99 puffer fish were collected from the coast of Seybaplaya, Campeche. Each fish had 20 µl of peripheral blood drawn, and blood smears were performed in triplicate. The smears were stained with Giemsa stain, and a quantitative analysis of blood cells (erythrocytes, leukocytes, and monocytes) was obtained with an optical microscope at 100 ×. The parasites recovered from each fish were fixed and identified, and the infection parameters were calculated. Through generalized additive model analysis (GAMLSS), we observed that the infection intensity of puffer fish influenced changes in hematological parameters, principally in erythrocytes, neutrophils, thrombocytes, the total fish length, and the condition factor of the fish. In conclusion, this is the first study that provides baseline data on the hematological parameter variations in uninfected and infected L. laevigatus, the tropical wild puffer fish, as well as the possible effects on fish health. It is necessary to establish reference hematological patterns in wild populations for diagnosis and timely management with emphasis on aquaculture fish.


Fetal alcohol syndrome: does alcohol withdrawal play a role?
Alcohol use by a pregnant woman may interfere with the development of her fetus. Newborns whose mothers are intoxicated during delivery can experience withdrawal symptoms, such as tremors and even seizures. It is likely that withdrawal also can occur during fetal development. Thus, the possibility exists that withdrawal by the pregnant woman may exacerbate alcohol's adverse effects on her fetus. One potential mechanism through which alcohol withdrawal might damage the fetus involves the receptor for the neurotransmitter glutamate (i.e., the N-methyl-D-aspartate [NMDA] receptor). This receptor plays a crucial role during neuronal development. Excessive activation of the NMDA receptor, which occurs during withdrawal, may lead to neuronal cell death. Animal studies suggest that these effects may contribute to behavioral deficits following prenatal exposure to alcohol.


Mechanism-based inactivation of cytochrome P450 3A4 by lapatinib.
Fatalities stemming from hepatotoxicity associated with the clinical use of lapatinib (Tykerb), an oral dual tyrosine kinase inhibitor (ErbB-1 and ErbB-2) used in the treatment of metastatic breast cancer, have been reported. We investigated the inhibition of CYP3A4 by lapatinib as a possible cause of its idiosyncratic toxicity. Inhibition of CYP3A4 was time-, concentration-, and NADPH-dependent, with k(inact) = 0.0202 min(-1) and K(i) = 1.709 μM. The partition ratio was approximately 50.9. Addition of GSH did not affect the rate of inactivation. Testosterone protected CYP3A4 from inactivation by lapatinib. The characteristic Soret peak associated with a metabolite-intermediate complex was not observed for lapatinib during spectral difference scanning. However, reduced carbon monoxide (CO)-difference spectroscopy did reveal a 43% loss of the spectrally detectable CYP3A4-CO complex in the presence of lapatinib. Incubation of either lapatinib or its dealkylated metabolite with human liver microsomes in the presence of GSH resulted in the formation of a reactive metabolite (RM)-GSH adduct derived from the O-dealkylated metabolite of lapatinib. In addition, coincubation of lapatinib with ketoconazole inhibited the formation of the RM-GSH adduct. In conclusion, we demonstrated for the first time that lapatinib is a mechanism-based inactivator of CYP3A4, most likely via the formation and further oxidation of its O-dealkylated metabolite to a quinoneimine that covalently modifies the CYP3A4 apoprotein and/or heme moiety.


Antiviral memory CD8 T-cell differentiation, maintenance, and secondary expansion occur independently of MyD88.
Inflammatory signals induced during infection regulate T-cell expansion, differentiation, and memory formation. Toll-like receptors (TLRs) are inflammatory mediators that allow innate immune cells to recognize and respond to invading pathogens. In addition to their role in innate immune cells, we have found that signals delivered through the TLR adapter protein myeloid differentiation protein 88 (MyD88) play a critical, T cell-intrinsic role in supporting the survival and accumulation of antigen-specific effector cells after acute viral infection. However, the importance of MyD88-dependent signals in regulating the generation and maintenance of memory T cells remained unclear. To address this, we used a novel, inducible knockout system to examine whether MyD88 is required for optimal memory CD8 T-cell generation and responses after lymphocytic choriomeningitis virus infection. We show that whereas MyD88 is critical for initial T-cell expansion, it is not required for the subsequent differentiation and stable maintenance of a memory T-cell population. Furthermore, in contrast to naive CD8 T cells, memory CD8 T cells do not depend on MyD88 for their secondary expansion. Our findings clarify the importance of MyD88 during distinct phases of the antiviral T-cell response and establish differential dependence on MyD88 signaling as a novel characteristic that distinguishes naive from memory CD8 T cells.


Circular DNA logic gates with strand displacement.
Circular DNA logic gates were constructed on the basis of DNA three-way branch migration. In this logic system, circular DNA was used as a basic work unit and linear single-strand DNA was used as input and output signals. Making use of the circular structure, most of the DNA-specific recognition regions were designed in a single DNA ring. Depending on accurate DNA sequence recognition and highly effective strand displacement, the logic gates yielded correct results. In addition, the positions of gold nanoparticles (AuNPs) were detected as an alternative approach to determine logic results. Thus, the accurate and tunable control of DNA/AuNPs may be applied widely in DNA nanotechnology.


The long-term outcome of microvascular decompression for trigeminal neuralgia.
Several surgical procedures to treat trigeminal neuralgia (tic douloureux) are available, but most reports provide only short-term follow-up information. We describe the long-term results of surgery in 1185 patients who underwent microvascular decompression of the trigeminal nerve for medically intractable trigeminal neuralgia. The outcome of the procedure was assessed prospectively with annual questionnaires. Of the 1185 patients who underwent microvascular decompression during the 20-year study period, 1155 were followed for 1 year or more after the operation. The median follow-up period was 6.2 years. Most postoperative recurrences of tic took place in the first two years after surgery. Thirty percent of the patients had recurrences of tic during the study period, and 11 percent underwent second operations for the recurrences. Ten years after surgery, 70 percent of the patients (as determined by Kaplan-Meier analysis) had excellent final results-that is, they were free of pain without medication for tic. An additional 4 percent had occasional pain that did not require long-term medication. Ten years after the procedure, the annual rate of the recurrence of tic was less than 1 percent. Female sex, symptoms lasting more than eight years, venous compression of the trigeminal-root entry zone, and the lack of immediate postoperative cessation of tic were significant predictors of eventual recurrence. Having undergone a previous ablative procedure did not lessen a patient's likelihood of having a cessation of tic after microvascular decompression, but the rates of burning and aching facial pain, as reported on the last follow-up questionnaire, were higher if a trigeminal-ganglion lesion had been created with radiofrequency current before microvascular decompression. Major complications included two deaths shortly after the operation (0.2 percent) and one brain-stem infarction (0.1 percent). Sixteen patients (1 percent) had ipsilateral hearing loss. Microvascular decompression is a safe and effective treatment for trigeminal neuralgia, with a high rate of long-term success.


Natural and anthropogenic radionuclide activity concentrations in the New Zealand diet.
To support New Zealand's food safety monitoring regime, a survey was undertaken to establish radionuclide activity concentrations across the New Zealand diet. This survey was undertaken to better understand the radioactivity content of the modern diet and also to assess the suitability of the current use of milk as a sentinel for dietary radionuclide trends. Thirteen radionuclides were analysed in 40 common food commodities, including animal products, fruits, vegetables, cereal grains and seafood. Activity was detected for (137)Caesium, (90)Strontium and (131)Iodine. No other anthropogenic radionuclides were detected. Activity concentrations of the three natural radionuclides of Uranium and the daughter radionuclide (210)Polonium were detected in the majority of food sampled, with a large variation in magnitude. The maximum activity concentrations were detected in shellfish for all these radionuclides. Based on the established activity concentrations and ranges, the New Zealand diet contains activity concentrations of anthropogenic radionuclides far below the Codex Alimentarius guideline levels. Activity concentrations obtained for milk support its continued use as a sentinel for monitoring fallout radionuclides in terrestrial agriculture. The significant levels of natural and anthropogenic radionuclide activity concentrations detected in finfish and molluscs support undertaking further research to identify a suitable sentinel for New Zealand seafood monitoring.