In vitro resistance of Enterobacter cloacae isolated from fresh seafood to colistin.
Enterobacter cloacae is a clinically important bacterium from the Enterobacteriaceae family. This study evaluated resistance of E. cloacae strains from fish (n=14) and shrimp (n=9) to colistin. Biochemical identification and antimicrobial susceptibility tests were carried out in an automated Vitek®2 instrument. Colistin resistance was observed in 21.4% and 66.7% of the strains from fish and shrimp, respectively. We observed minimum inhibitory concentrations of ≥16 mg/L and ≤5 mg/L in 8 and 15 of all strains, respectively. Fish and shrimp can carry drug-resistant enterobacteria, which can be of clinical interest.


Jets, stickiness, and anomalous transport.
Dynamical and statistical properties of the vortex and passive particle advection in chaotic flows generated by 4- and 16-point vortices are investigated. General transport properties of these flows are found to be anomalous and exhibit a superdiffusive behavior with typical second moment exponent mu approximately 1.75. The origin of this anomaly is traced to the presence of coherent structures within the flow, the vortex cores, and the region far from where vortices are located. In the vicinity of these regions stickiness is observed and the motion of tracers is quasiballistic. The chaotic nature of the underlying flow dictates the choice for thorough analysis of transport properties. Passive tracer motion is analyzed by measuring the mutual relative evolution of two nearby tracers. Some tracers travel in each other's vicinity for relatively long times. This is related to a hidden order for the tracers, which we call jets. Jets are localized and found in sticky regions. Their structure is analyzed and found to be formed of a nested set of jets within jets. The analysis of the jet trapping time statistics shows a quantitative agreement with the observed transport exponent.


[Investigation of erythromycin resistance and resistance phenotypes in group A beta hemolytic streptococci].
Although penicillin resistance has not been determined in group A beta haemolytic streptococci (GABHS) yet, resistance to erythromycin and other macrolids is being reported frequently in the last years. In this study we investigated erythromycin resistance by using agar dilution method in 282 GABHS strains which were isolated from throat cultures that had been evaluated in the Ministry of Health, Ankara Training and Research Hospital's, Microbiology Laboratory. We also determined resistance phenotypes of resistant strains by double disc synergy method using erythromycin and clindamycin discs. Twelve of 282 strains (4.3%) were found resistant to erythromycin; five (41.7%) of which were M phenotype, four (33.3%) of which were constitutive type MLSB phenotype and three (25%) of which were inducible type MLSB phenotype. Investigation of resistance to macrolides that are alternatives to penicillin therapy in GABHS, is very important for the determination of the therapy and also to provide epidemiological data.


Embryonic development of the nucleus isthmo-opticus in the chick: a Golgi and electron microscopic study.
The cytology and the synaptology of the nucleus isthmo-opticus have been studied in the chick embryo at three stages during (incubation days 14 and 15) and after (day 18) the neurone death period of the nucleus. The maturation process is not synchronous throughout the nucleus:the dorsal most region lags behind the rest of the nucleus. On the other hand, cell degeneration is equally distributed over the nucleus at days 14 and 15. At day 14, the nucleus shows little cellular alignment. The neuropil is stocked with immature neurites. Synaptogenesis essentially concerns axodendritic contacts, and the presynaptic partners contain only round vesicles. A few axons undergo myelination; among them, some are degenerating. At day 15, cellular alignment is much more pronounced. Synaptogenesis is very active; it concerns axosomatic as well as axodendritic contacts. A majority of the presynaptic bags contain only round vesicles, although some have a pleomorphic population of vesicles. At day 18, the histological structure of the nucleus is similar to that found in the adult. Although the neuropil presents mature profiles (development of dendritic spines, presence of synaptic glomeruli), it still contains numerous immature features (active synaptogenesis, persistent growth cones). Possible functional implications of this maturation are discussed. In particular, the fact that the nuclear neuropil is not fully developed by the end of the incubation period suggests that the accomplishment of the feedback role of the nucleus is determined by function.


The interactome of metabolic enzyme carbonic anhydrase IX reveals novel roles in tumor cell migration and invadopodia/MMP14-mediated invasion.
Carbonic anhydrase IX (CAIX) is a hypoxia inducible factor 1-induced, cell surface pH regulating enzyme with an established role in tumor progression and clinical outcome. However, the molecular basis of CAIX-mediated tumor progression remains unclear. Here, we have utilized proximity dependent biotinylation (BioID) to map the CAIX 'interactome' in breast cancer cells in order to identify physiologically relevant CAIX-associating proteins with potential roles in tumor progression. High confidence proteins identified include metabolic transporters, β1 integrins, integrin-associated protein CD98hc and matrix metalloprotease 14 (MMP14). Biochemical studies validate the association of CAIX with α2β1 integrin, CD98hc and MMP14, and immunofluorescence microscopy demonstrates colocalization of CAIX with α2β1 integrin and MMP14 in F-actin/cofilin-positive lamellipodia/pseudopodia, and with MMP14 to cortactin/Tks5-positive invadopodia. Modulation of CAIX expression and activity results in significant changes in cell migration, collagen degradation and invasion. Mechanistically, we demonstrate that CAIX associates with MMP14 through potential phosphorylation residues within its intracellular domain, and that CAIX enhances MMP14-mediated collagen degradation by directly contributing hydrogen ions required for MMP14 catalytic activity. These findings establish hypoxia-induced CAIX as a novel metabolic component of cellular migration and invasion structures, and provide new mechanistic insights into its role in tumor cell biology.


Compatibility and Stability of Rolapitant Injectable Emulsion Admixed with Dexamethasone Sodium Phosphate.
Neurokinin-1 receptor antagonist, 5-hydroxytryptamine-3 receptor antagonist, and dexamethasone combination therapy is the standard of care for the prevention of chemotherapy-induced nausea and vomiting. Herein, we describe the physical and chemical stability of an injectable emulsion of the Neurokinin-1 receptor antagonist rolapitant 185 mg in 92.5 mL (free base, 166.5 mg in 92.5 mL) admixed with either 2.5 mL of dexamethasone sodium phosphate (10 mg) or 5 mL of dexamethasone sodium phosphate (20 mg). Admixtures were prepared and stored in two types of container closures (glass and Crystal Zenith plastic bottles) and four types of intravenous administration tubing sets (or intravenous tubing sets). The assessment of the physical and chemical stability was conducted on admixtures packaged in bottled samples stored at room temperature (20°C to 25°C under fluorescent light) and evaluated at 0, 1, and 6 hours. For admixtures in intravenous tubing sets, the assessment of physicochemical stability was performed after 0 and 7 hours of storage at 20°C to 25°C, and then after 20 hours (total 27 hours) under refrigeration (2°C to 8°C) and protected from light. Physical stability was assessed by visually examining the bottle contents under normal room light and measuring turbidity and particulate matter. Chemical stability was assessed by measuring the pH of the admixture and determining drug concentrations through high-performance liquid chromatographic analysis. Results showed that all samples were physically compatible throughout the duration of the study. The admixtures stayed within narrow and acceptable ranges in pH, turbidity, and particulate matter. Admixtures of rolapitant and dexamethasone were chemically stable when stored in glass and Crystal Zenith bottles for at least 6 hours at room temperature, as well as in the four selected intravenous tubing sets for 7 hours at 20°C to 25°C and then for 20 (total 27 hours) hours at 2°C to 8°C. No loss of potency of any admixed component occurred in the samples stored at the temperature ranges studied.


Effect of barley and oat cultivars with different carbohydrate compositions on the intestinal bacterial communities in weaned piglets.
This experiment was aimed at comparing the intestinal microbial community composition in pigs fed hulled common barley supplemented with isolated barley mixed-linked beta-glucan, four hulless barley varieties and breeding lines with mixed-linked beta-glucan contents ranging from 41 to 84 g kg(-1) and different amylose/amylopectin ratios as well as two oat varieties. Seventy-two weaned piglets were allocated to one of nine diets composed of 81.5% cereal, 6% whey, 9% soy protein isolate and 3.5% minerals. After 15 days, pigs were sacrificed and ileum and colon contents were collected for quantitative real-time PCR (qPCR) and denaturing gradient gel electrophoresis to evaluate microbial communities. Shifts in intestinal microbial communities were observed with the hulless barley cultivars with a normal to high beta-glucan content and from normal starch toward either high-amylopectin or high-amylose starch. These hulless barleys had the lowest (P<0.05) microbial diversity, whereas oats had intermediate diversity compared with low-beta-glucan hulless cultivars and hulled varieties. Furthermore, hulless varieties favoured xylan- and beta-glucan-degrading bacteria whereas mixed-linked beta-glucan-supplemented hulled barley favoured lactobacilli. Numbers of lactobacilli decreased in the ileum of pigs fed hulless/high mixed-linked beta-glucan barley-based diets. Thus, cultivar differences in both the form and the quantity of carbohydrates affect gut microbiota in pigs, which provides information for future feeding strategies.


[Possibilities for using emergency abdominal ultrasound study and its significance in acute diseases of the abdominal cavity].
Studies were conducted by the authors' of this paper into the diagnostic value of ultrasonic examination to clear up suspicion of acute abdominal problems, with 381 examinations being actually evaluated. Definitely valid diagnosis was obtained from the method in 37.7 per cent of these cases. The tentative diagnosis was invalidated in 18.2 per cent. The results are compared with literature data. Continued systematic use of the approach is recommended. It would bring down the time required for diagnosis and might provide another possibility of supporting the diagnostic effort which is quite difficult, when it comes to acute abdominal problems.


γδ T Cell-Mediated Immune Responses in Disease and Therapy.
The role of γδ T cells in immunotherapy has gained specific importance in the recent years because of their prominent function involving directly or indirectly in the rehabilitation of the diseases. γδ T cells represent a minor population of T cells that express a distinct T cell receptor (TCR) composed of γδ chains instead of αβ chains. Unlike αβ T cells, γδ T cells display a restricted TCR repertoire and recognize mostly unknown non-peptide antigens. γδ T cells act as a link between innate and adaptive immunity, because they lack precise major histocompatibility complex (MHC) restriction and seize the ability to recognize ligands that are generated during affliction. Skin epidermal γδ T cells recognize antigen expressed by damaged or stressed keratinocytes and play an indispensable role in tissue homeostasis and repair through secretion of distinct growth factors. γδ T cell based immunotherapy strategies possess great prominence in the treatment because of the property of their MHC-independent cytotoxicity, copious amount of cytokine release, and a immediate response in infections. Understanding the role of γδ T cells in pathogenic infections, wound healing, autoimmune diseases, and cancer might provide knowledge for the successful treatment of these diseases using γδ T cell based immunotherapy. Enhancing the human Vγ9Vδ2 T cells functions by administration of aminobisphosphonates like zoledronate, pamidronate, and bromohydrin pyrophosphate along with cytokines and monoclonal antibodies shows a hopeful approach for treatment of tumors and infections. The current review summarizes the role of γδ T cells in various human diseases and immunotherapeutic approaches using γδ T cells.


CTLA4 Promotes Tyk2-STAT3-Dependent B-cell Oncogenicity.
CTL-associated antigen 4 (CTLA4) is a well-established immune checkpoint for antitumor immune responses. The protumorigenic function of CTLA4 is believed to be limited to T-cell inhibition by countering the activity of the T-cell costimulating receptor CD28. However, as we demonstrate here, there are two additional roles for CTLA4 in cancer, including via CTLA4 overexpression in diverse B-cell lymphomas and in melanoma-associated B cells. CTLA4-CD86 ligation recruited and activated the JAK family member Tyk2, resulting in STAT3 activation and expression of genes critical for cancer immunosuppression and tumor growth and survival. CTLA4 activation resulted in lymphoma cell proliferation and tumor growth, whereas silencing or antibody-blockade of CTLA4 in B-cell lymphoma tumor cells in the absence of T cells inhibits tumor growth. This inhibition was accompanied by reduction of Tyk2/STAT3 activity, tumor cell proliferation, and induction of tumor cell apoptosis. The CTLA4-Tyk2-STAT3 signal pathway was also active in tumor-associated nonmalignant B cells in mouse models of melanoma and lymphoma. Overall, our results show how CTLA4-induced immune suppression occurs primarily via an intrinsic STAT3 pathway and that CTLA4 is critical for B-cell lymphoma proliferation and survival. Cancer Res; 77(18); 5118-28. ©2017 AACR.


Effect of acute ozone exposure on the lung metabolomes of obese and lean mice.
Pulmonary responses to the air pollutant, ozone, are increased in obesity. Both obesity and ozone cause changes in systemic metabolism. Consequently, we examined the impact of ozone on the lung metabolomes of obese and lean mice. Lean wildtype and obese db/db mice were exposed to acute ozone (2 ppm for 3 h) or air. 24 hours later, the lungs were excised, flushed with PBS to remove blood and analyzed via liquid-chromatography or gas-chromatography coupled to mass spectrometry for metabolites. Both obesity and ozone caused changes in the lung metabolome. Of 321 compounds identified, 101 were significantly impacted by obesity in air-exposed mice. These included biochemicals related to carbohydrate and lipid metabolism, which were each increased in lungs of obese versus lean mice. These metabolite changes may be of functional importance given the signaling capacity of these moieties. Ozone differentially affected the lung metabolome in obese versus lean mice. For example, almost all phosphocholine-containing lysolipids were significantly reduced in lean mice, but this effect was attenuated in obese mice. Glutathione metabolism was also differentially affected by ozone in obese and lean mice. Finally, the lung metabolome indicated a role for the microbiome in the effects of both obesity and ozone: all measured bacterial/mammalian co-metabolites were significantly affected by obesity and/or ozone. Thus, metabolic derangements in obesity appear to impact the response to ozone.


In-situ investigation of the adsorption of globular model proteins on stimuli-responsive binary polyelectrolyte brushes.
Binary brushes constituted from two incompatible polymers can be used in the form of ultrathin polymeric layers as a versatile tool for surface engineering to tune physicochemical surface characteristics such as wettability, surface charge, chemical composition, and morphology and furthermore to create responsive surface properties. Mixed brushes of oppositely charged weak polyelectrolytes represent a special case of responding surfaces that are sensitive to changes in the pH value of the aqueous environment and therefore represent interesting tools for biosurface engineering. The polyelectrolyte brushes used for this study were composed of two oppositely charged polyelelctrolytes poly(2-vinylpyridine) (P2VP) and poly(acrylic acid) (PAA). The in-situ properties and surface characteristics such as as surface charge, surface tension, and extent of swelling of these brush layers are functions of the pH value of the surrounding aqueous solution. To test the behavior of the mixed polylelctrolyte brushes in contact with biosystems, protein adsorption experiments with globular model proteins were performed at different pH values and salt concentrations (confinement of counterions) of the buffer solutions. The influence of the pH value, buffer salt concentration, and isoelectric points (IEP) of the brush and protein on the adsorbed amount and the interfacial tension during protein adsorption as well as the protein adsorption mechanism postulated in reference to recently developed theories of protein adsorption on polyelectrolyte brushes is discussed. In the salted regime, protein adsorption was found to be similar to the often-described adsorption at hydrophobic surfaces. However, in the osmotic regime the balance of electrostatic repulsion and a strong entropic driving force, "counterion release", was found to be the main influence on protein adsorption.


Oxygen activation at mononuclear nonheme iron centers: a superoxo perspective.
Dioxygen (O(2)) activation by iron enzymes is responsible for many metabolically important transformations in biology. Often a high-valent iron oxo oxidant is proposed to form upon O(2) activation at a mononuclear nonheme iron center, presumably via intervening iron superoxo and iron peroxo species. While iron(IV) oxo intermediates have been trapped and characterized in enzymes and models, less is known of the putative iron(III) superoxo species. Utilizing a synthetic model for the 2-oxoglutarate-dependent monoiron enzymes, [(Tp(iPr2))Fe(II)(O(2)CC(O)CH(3))], we have obtained indirect evidence for the formation of the putative iron(III) superoxo species, which can undergo one-electron reduction, hydrogen-atom transfer, or conversion to an iron(IV) oxo species, depending on the reaction conditions. These results demonstrate the various roles that the iron(III) superoxo species can play in the course of O(2) activation at a nonheme iron center.


Identification of mouse mammary epithelial cells by immunofluorescence with rabbit and guinea pig antikeratin antisera.
Few markers are available to identify the three types of mammary epithelial cells--ductal, alveolar, and myoepithelial--especially in pathological conditions and in cell cultures. We have used antisera to human keratins in immunofluorescence to facilitate the identification of the three mouse mammary epithelial cell types. In frozen tissue sections and primary cell cultures, a rabbit antikeratin antiserum specifically stained cytoplasmic filaments in all three types of epithelial cells. A guinea pig antiserum against the same keratin preparation, however, reacted preferentially with filaments in myoepithelial cells and readily detected this cell type in normal, dysplastic, and malignant mammary tissues and cell cultures. Neither antisera reacted with fibroblasts or any other mesenchymal cells. The combined use of the two antikeratin antisera thereby permits rapid surveys of tissue sections and cultures for the localization of not only all epithelial cells but also the subpopulation of myoepithelial cells. Moreover, when mammary cultures established from late-pregnant or lactating mice were stained simultaneously with guinea pig antikeratin and rabbit anticasein antisera, three populations of epithelial cells were mutually exclusive: those stained by anticasein antiserum, those stained by guinea pig antikeratin antiserum, and those stained by neither, consistent with properties of alveolar, myoepithelial, and ductal cells, respectively. These antisera thus offer a tool for studying different epithelial cell types during mammary development, tumorigenesis, and malignant progression.


A Caffeic Acid Matrix Improves In Situ Detection and Imaging of Proteins with High Molecular Weight Close to 200,000 Da in Tissues by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging.
To our knowledge, this was the first study in which caffeic acid (CA) was successfully evaluated as a matrix to enhance the in situ detection and imaging of endogenous proteins in three biological tissue sections (i.e., a rat brain and Capparis masaikai and germinating soybean seeds) by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). Our results show several properties of CA, including strong ultraviolet absorption, a super-wide MS detection mass range close to 200,000 Da, micrometer-sized matrix crystals, uniform matrix deposition, and high ionization efficiency. More high-molecular-weight (HMW) protein ion signals (m/z > 30,000) could be clearly detected in biological tissues with the use of CA, compared to two commonly used MALDI matrices, i.e., sinapinic acid (SA) and ferulic acid (FA). Notably, CA shows excellent performance for HMW protein in situ detection from biological tissues in the mass range m/z > 80,000, compared to the use of SA and FA. Furthermore, the use of a CA matrix also significantly enhanced the imaging of proteins on the surface of selected biological tissue sections. Three HMW protein ion signals (m/z 50,419, m/z 65,874, and m/z 191,872) from a rat brain, two sweet proteins (mabinlin-2 and mabinlin-4) from a Capparis masaikai seed, and three HMW protein ion signals (m/z 94,838, m/z 134,204, and m/z 198,738) from a germinating soybean seed were successfully imaged for the first time. Our study proves that CA has the potential to become a standard organic acid matrix for enhanced tissue imaging of HMW proteins by MALDI-MSI in both animal and plant tissues.


Use of drugs of abuse in less than 30-year-old drivers killed in a road crash in France: a spectacular increase for cannabis, cocaine and amphetamines.
A collaborative study was conducted in France in order to determine the prevalence of cannabinoids, opiates, cocaine metabolites and amphetamines in blood samples from drivers killed in road accidents in 2003 and 2004 and to compare these values with those of a previous study performed during the period 2000-2001 involving 900 drivers. Blood samples were provided from 2003 under 30-year-old drivers, killed in a traffic accident. Drugs of abuse were determined by gas chromatography-mass spectrometry using the same analytical procedures in all the 12 laboratories. The most frequently observed compounds were by far cannabinoids, that tested positive in 39.6% of the total number of samples. Delta9 tetrahydrocannabinol (THC), the most active of the principle constituents in marijuana (cannabis sativa), was detected in the blood of 28.9% drivers and was the single drug of abuse in 80.2% of the positive cases. It was associated with amphetamines in 7.4% and with opiates and cocaine in 1.9 and 4.8%, respectively. Amphetamines were present in 3.1% of the total number of samples, cocaine metabolites in 3.0% and opiates in 3.5%. When comparing these results with those of a previous study performed 3 years before, a significant increase is observed for THC (28.9% versus 16.9%), cocaine metabolites (3.0% versus 0.2%) and amphetamines (3.1% versus 1.4%). This study demonstrates the critical necessity of implementing in France as soon as possible systematical roadside testing for drugs of abuse.


Idiotypic analysis of hybridoma antibodies to branched synthetic polymer (Tyr, Glu)-Ala-Lys: idiotypic relationship with antibodies to linear random polymer (Glu60, Ala30, Tyr10).
The expression of three anti-GAT idiotypes, CGAT, Gte, and GA-1, on 17 C57BL/10 and four C3H.SW hybridoma anti-(T,G)-A--L antibodies was analyzed. These hybridoma anti-(T,G)-A--L antibodies exhibited two patterns of fine antigen binding specificity. The majority of the hybridoma antibodies bound the (T,G)-A--L, GT, and GAT polymers but not the GA polymer, and were designated as GT-reactive hybridoma antibodies. A minor population of hybridoma anti(T,G)-A--L antibodies bound to (T,G)-A--L but not to GT, GAT, or GA, i.e., (T,G)-A--L-specific. A complete correlation between fine antigen binding pattern and the expression of CGAT idiotype was demonstrated. None of the 21 hybridoma anti-(T,G)-A--L antibodies expressed the GA-1 idiotype. All of the GT-reactive and none of the GT-nonreactive hybridoma anti-(%,G)-A--L antibodies expressed the CGAT idiotype. Furthermore, the Gte idiotype was found on the majority of CGAT+-bearing C57BL/10 hybridoma anti-(T,G)-A--L antibodies. These results indicate that C57BL/10 anti-(T,G)-A--L antibody repertoire can be grouped into a minimum of three families; i.e., CGAT+ Gte+, CGAT+ Gte-, and CGAT- Gte- families, with the CGAT+ Gte+ family as the major compartment. This is confirmed by the high percentage idiotype binding of serum anti-(T,G)-A--L antibodies with anti-CGAT idiotypic antisera. Finally, anti-idiotypic antisera made against CGAT+ hybridoma anti-GAT or anti-(T,G)-A--L antibodies crossreact extensively with other CGAT+ hybridoma anti-GAT and anti-(T,G)-A--L antibodies. However, additional experiments demonstrated that CGAT+ hybridoma anti-(T,G)-A--L antibodies also possess private idiotypes.


Sandflies in the Greater Athens region, Greece.
During the period from May to October 1992, a survey of phlebotomine sandflies was conducted in the Greater Athens region where cases of human and canine leishmaniasis occur. Using castor-oil paper traps, a total of 3015 sandflies were caught. Of the identified specimens, 1002 (34.1%) were Phlebotomus neglectus, 541 (18.4%) were P. papatasi, 182 (6.2%) were P. tobbi, 50 (1.7%) were P. simici, 30 (1.0%) were P. alexandri, 3(0.4%) were P. sergenti and 1122 (38.2%) were Sergentomyia minuta. The seasonal activity of most Phlebotomus species was bimodal, while that of S. minuta was unimodal. The population density of sandflies varied significantly in different localities; it was low in most of the stations and high in some stations in the outskirts. Among the potential vectors of Leishmania spp., P. neglectus and P. Papatasi were the most widespread species. The former species showed high density in 12 of the 70 sampled stations, while the latter in only 4. Hence, the risk of transmission of sandfly-borne diseases is still present in the Greater Athens region.


Is there an effect of menopause on CHD mortality? Asia Pacific comparisons.
This study examines the mortality rates of men and women from coronary heart disease (CHD) in a number of Asia-Pacific countries with very different incidences of this disorder. In all countries, mortality rates from CHD were higher in men. In women, no unequivocal change in mortality rate from CHD attributable to the menopause could be detected.


Evaluation of school children at high risk for fragile X syndrome utilizing buccal cell FMR-1 testing.
We describe a pilot project utilizing saliva to identify the FMR-1 mutation in high-risk special education students from four public school districts in Colorado. The program included presentations to special education teachers regarding fragile X syndrome, parental consent for testing, completion of a behavior checklist by the teachers, identification of special education students at high risk for fragile X syndrome, subsequent brief examination of face and hands, collection of a saliva sample by either Gatorade swish or brushing of the inside of the cheek, and analysis for the FMR-1 mutation by PCR. Equivocal samples were studied by direct DNA testing using Southern blot analysis, and abnormal results were confirmed by a blood analysis for the FMR-1 mutation. Mutant individuals received genetic counseling and medical and educational assessments to optimize treatment and intervention. This pilot project was met with enthusiasm by the schools. Of the first 439 students evaluated, 68% were male with an average age of 7.75 years; 13% were mentally retarded or autistic. Most students referred for the evaluation were learning disabled (51%) and/or had an Attention Deficit Hyperactivity Disorder (ADHD) (35%). The overall prevalence of the FMR-1 mutation was 5 of 439 or 1.1%. This relatively low yield is probably due to the high number of non-retarded but learning disabled students tested. Of the mentally retarded patients tested, 3.5% were positive for the FMR-1 mutation; however, of the non-retarded or learning disabled patients, only 0.79% were FMR-1 positive.(ABSTRACT TRUNCATED AT 250 WORDS)


Painful rib syndrome--a review of 76 cases.
The painful rib syndrome consists of three features: pain in the lower chest or upper abdomen, a tender spot on the costal margin, and reproduction of the pain on pressing the tender spot. This is a common cause for referral to a general medical/gastroenterology clinic, accounting for 3% of new referrals in Lincoln. Seventy six consecutive patients were studied. The mean age was 48 years and 70% were women. Forty three per cent had been investigated, often extensively, before referral, and eight had had a non-curative cholecystectomy. The case notes from all patients were reviewed and a follow up questionnaire was sent after a mean period of four years to those 72 still alive, of which 56 replied. Thirty nine (70%) still had the pain although all except three had learnt to live with it. Despite a firm diagnosis being given, 25 (33%) patients were referred again to hospital by their general practitioner. All further investigations were negative apart from the finding of gall stones in three patients. The four patients who died had died from unrelated causes. The painful rib syndrome is common but underdiagnosed. It is a safe, clinical diagnosis requiring no investigation. Systematic firm palpation of the costal margin in recommended in all patients presenting with pain in the lower chest or upper abdomen.


Short-term oral α-lipoic acid does not prevent lipid-induced dysregulation of glucose homeostasis in obese and overweight nondiabetic men.
Prolonged elevation of plasma free fatty acids (FFAs) induces insulin resistance and impairs pancreatic β-cell adaptation to insulin resistance. The mechanisms whereby lipid induces these impairments are not fully defined but may involve oxidative stress, inflammation, and endoplasmic reticulum stress. α-Lipoic acid (ALA), a commonly used health supplement with antioxidant, anti-inflammatory, and AMPK-activating properties, has been shown to have therapeutic value in type 2 diabetes and its complications. Here we examined the effects of ALA on insulin sensitivity and secretion in humans under the conditions of 24-h iv lipid infusion to elevate plasma FFAs. Eight overweight and obese male subjects underwent four randomized studies each, 4-6 wk apart: 1) SAL, 2-wk oral placebo followed by 24-h iv infusion of saline; 2) IH, 2-wk placebo followed by 24-h iv infusion of intralipid plus heparin to raise plasma FFAs approximately twofold; 3) IH + ALA, 2-wk ALA (1,800 mg/day) followed by 24-h infusion of intralipid plus heparin; and 4) ALA, 2-wk ALA followed by 24-h infusion of saline. Insulin secretion rates (ISR) and insulin sensitivity were assessed with a 2-h, 20-mmol/l hyperglycemic clamp and a hyperinsulinemic euglycemic clamp, respectively. ISR was not significantly different between treatments. Lipid infusion impaired insulin sensitivity with and without ALA pretreatment. These results indicate that ALA, administered orally at this dose for 2 wk, does not protect against lipid-induced insulin resistance in overweight and obese humans.


Application of real-time PCR to estimate toxin production by the cyanobacterium Planktothrix sp.
Quantitative real-time PCR methods are increasingly being applied for the enumeration of toxic cyanobacteria in the environment. However, to justify the use of real-time PCR quantification as a monitoring tool, significant correlations between genotype abundance and actual toxin concentrations are required. In the present study, we aimed to explain the concentrations of three structural variants of the hepatotoxin microcystin (MC) produced by the filamentous cyanobacterium Planktothrix sp., [Asp, butyric acid (Dhb)]-microcystin-RR (where RR means two arginines), [Asp, methyl-dehydro-alanine (Mdha)]-microcystin-RR, and [Asp, Dhb]-microcystin-homotyrosine-arginine (HtyR), by the abundance of the microcystin genotypes encoding their synthesis. Three genotypes of microcystin-producing cyanobacteria (denoted the Dhb, Mdha, and Hty genotypes) in 12 lakes of the Alps in Austria, Germany, and Switzerland from 2005 to 2007 were quantified by means of real-time PCR. Their absolute and relative abundances were related to the concentration of the microcystin structural variants in aliquots determined by high-performance liquid chromatography (HPLC). The total microcystin concentrations varied from 0 to 6.2 microg liter(-1) (mean +/- standard error [SE] of 0.6 +/- 0.1 microg liter(-1)) among the samples, in turn resulting in an average microcystin content in Planktothrix of 3.1 +/- 0.7 microg mm(-3) biovolume. Over a wide range of the population density (0.001 to 3.6 mm(3) liter(-1) Planktothrix biovolume), the Dhb genotype and [Asp, Dhb]-MC-RR were most abundant, while the Hty genotype and MC-HtyR were found to be in the lowest proportion only. In general, there was a significant linear relationship between the abundance/proportion of specific microcystin genotypes and the concentration/proportion of the respective microcystin structural variants on a logarithmic scale. We conclude that estimating the abundance of specific microcystin genotypes by quantitative real-time PCR is useful for predicting the concentration of microcystin variants in water.


Decolorization pathways of anthraquinone dye Disperse Blue 2BLN by Aspergillus sp. XJ-2 CGMCC12963.
Anthraquinone dye represents an important group of recalcitrant pollutants in dye wastewater. Aspergillus sp XJ-2 CGMCC12963 showed broad-spectrum decolorization ability, which could efficiently decolorize and degrade various anthraquinone dyes (50 mg L-1) under microaerophilic condition. And the decolorization rate of 93.3% was achieved at 120 h with Disperse Blue 2BLN (the target dye). Intermediates of degradation were detected by FTIR and GC-MS, which revealed the cleavage of anthraquinone chromophoric group and partial mineralization of target dye. In addition, extracellular manganese peroxidase showed the most closely related to the increasing of decolorization rate and biomass among intracellular and extracellular ligninolytic enzymes. Given these results, 2 possible degraded pathways of target dye by Aspergillus sp XJ-2 CGMCC12963 were proposed first in this work. The degradation of Disperse Blue 2BLN and broad spectrum decolorization ability provided the potential for Aspergillus sp XJ-2 CGMCC12963 in the treatment of wastewater containing anthraquinone dyes.


Voltage imaging of waking mouse cortex reveals emergence of critical neuronal dynamics.
Complex cognitive processes require neuronal activity to be coordinated across multiple scales, ranging from local microcircuits to cortex-wide networks. However, multiscale cortical dynamics are not well understood because few experimental approaches have provided sufficient support for hypotheses involving multiscale interactions. To address these limitations, we used, in experiments involving mice, genetically encoded voltage indicator imaging, which measures cortex-wide electrical activity at high spatiotemporal resolution. Here we show that, as mice recovered from anesthesia, scale-invariant spatiotemporal patterns of neuronal activity gradually emerge. We show for the first time that this scale-invariant activity spans four orders of magnitude in awake mice. In contrast, we found that the cortical dynamics of anesthetized mice were not scale invariant. Our results bridge empirical evidence from disparate scales and support theoretical predictions that the awake cortex operates in a dynamical regime known as criticality. The criticality hypothesis predicts that small-scale cortical dynamics are governed by the same principles as those governing larger-scale dynamics. Importantly, these scale-invariant principles also optimize certain aspects of information processing. Our results suggest that during the emergence from anesthesia, criticality arises as information processing demands increase. We expect that, as measurement tools advance toward larger scales and greater resolution, the multiscale framework offered by criticality will continue to provide quantitative predictions and insight on how neurons, microcircuits, and large-scale networks are dynamically coordinated in the brain.


Mood in relation to subclinical obsessive-compulsive symptoms.
Obsessive-compulsive disorder (OCD) is considered an anxiety disorder, but shows comorbidity with other disorders in the affective and impulsive-compulsive spectra, including anxiety disorders, major depression, and drug addictions. Subclinical OCD symptoms are relatively common in nonclinical populations and share common neurobiological substrates with clinical OCD. In this nonclinical community sample, the relationship between the severity of obsessions and compulsions, as measured by the Yale-Brown Obsessive Compulsive Scale, related to the intensity of negative emotions (anger, depression, tension, confusion, and fatigue) but not positive emotion (vigor), as measured by the Profile of Mood States. These relationships were independent of demographic influences and psychoactive drug use frequency (alcohol, cannabis, opioid, major stimulants, MDMA, and hallucinogens). These likely reflect common neurobiological substrates for emotional and behavioral regulation in prefrontal-subcortical/limbic circuits, which show normal variations in the general population.


Justice-Involved Women and Cervical Cancer Disparities: Issues of Surveillance, Education, and Policy.
Incarceration affects an increasing number of women in the United States. For current and formerly incarcerated women (justice-involved women), incarceration has implications for health, particularly the reproductive health of women, long after incarceration is over. Currently, justice-involved women have a high cervical cancer burden relative to the general population, resulting in substantial disparities in incidence and outcomes. In this article, we review the surveillance, education, and resulting policy issues that contribute to incarceration being a determinant of cervical cancer disparities and present a potential model for continuity of care to reduce such inequity.


A preliminary report of patients treated with early loaded implants using computerized tomography-guided surgical stents: flapless versus conventional flapped surgery.
The objective of this clinical study was to compare the survival rates of early loaded implants placed using flapless and flapped surgical techniques and to determine the bone density in the implant recipient sites using computerized tomography (CT). The study population consisted of 12 patients who were referred implant placement. One group consisted of five patients referred for the placement of 14 implants and treated with a flapless procedure. The other group consisted of seven patients referred for the placement of 45 implants with a conventional flapped procedure. Patients were selected randomly. CT machine was used for pre-operative evaluation of the jaw bone and the mean bone density value of each implant recipient site was recorded in Hounsfield units (HU). All implants were placed using CT-guided surgical stents. The early loading protocols included 2 months of healing in the mandible and 3 months of healing in the maxilla. Single-implant crowns, implant-supported fixed partial dentures, and implant-retained over dentures were delivered to the patients. Of 59 implants placed, one was lost in the conventional flapped group within the first month of healing, meaning overall implant survival rate of 98.3% average 9 months later. The highest average bone density value (801 +/- 239 HU) was found in the anterior mandible, followed by 673 +/- 449 HU for the posterior maxilla, 669 +/-346 HU for the anterior maxilla and 538 +/- 271 HU for the posterior mandible. The results of this study show that the early loading of implants placed utilizing flapless surgical technique with CT-guided surgical stents may be possible.


Evidence for ammonium-bearing minerals on ceres.
Spectra obtained from recent telescopic observation of 1-Ceres and laboratory measurements and theoretical calculations of three component mixtures of Ceres analog material suggest that an ammoniated phyllosilicate is present on the surface of the asteroid, rather than H(2)O frost as had been previously reported. The presence of an ammoniated phyllosilicate, most likely ammoniated saponite, on the surface of Ceres implies that secondary temperatures could not have exceeded 400 kelvin.


Ulnar/fibular ray defect and brachydactyly in a family: a possible new autosomal dominant syndrome.
The ulnar-mammary syndrome (MIM 181450) includes postaxial ray defects, abnormalities of growth, delayed sexual development, and mammary and apocrine gland hypoplasia. Brachydactyly type E (MIM 113300) presents with shortening of the metacarpals and phalanges in the ulnar ray in association with moderately short stature. We describe a three-generation family with variable expression of ulnar/fibular hypoplasia, brachydactyly, ulnar ray defects and short stature. The proband had ulnar hypoplasia with missing IV-Vth fingers, fibular hypoplasia on the right, bilateral club feet, growth retardation, a hypoplastic mid-face, an ASD and hemangiomas. She had normal mammary tissue and normal sweating. The mother had short stature, midfacial hypoplasia, a hypoplastic ulna and hypoplasia of the IVth metacarpal (brachydactyly) on the right without other associated malformations. The maternal grandfather had mild bilateral fibular hypoplasia and midphalangeal brachydactyly of the IV-Vth toes. His sister had mild short stature and shortening of the IVth metacarpal of the left hand. Two-point linkage analysis with microsatellite markers spanning the Ulnar-Mammary locus at 12q24.1 did not confirm linkage. The patients may have a previously undescribed syndrome.


Components of successful HIV/AIDS case management in Alaska Native villages.
Case management is a valuable support service for HIV/AIDS patients, but little is known about the elements of case management that contribute to success for Alaska Natives living in remote rural villages. This study used qualitative data from 18 key informants (social services/health workers, family members, and tribal leaders) to identify the elements of successful case management. Two major categories dominated the analysis. First, the case management organization possessed attributes, such as location, coordination, and support for professionals, that contributed to a positive case management experience. Second, the case manager had both knowledge/professional skills and values with personal capabilities that contributed to positive experiences. These components of successful case management can be integrated into existing and future case management programs for Alaska Natives to improve client level outcomes.


[Study of anti-macrophage serum and of its effect on antibody formation].
A specific anti-macrophage serum (AMS) has been obtained by immunizing rabbits with mouse peritoneal exudate cells and after repeated absorption with mouse erythrocytes, thymocytes and splenic lymphocytes. A comparative study of the cytotoxic activities of this serum on macrophages, thymocytes and lymph node cells, before and after absorption, showed that macrophage posses antigens which are common to both thymic and lymph node lymphocytes, and other antigens which are in common with lymph node lymphocytes but absent on thymocytes, as well as antigens being particular to macrophages. The absorbed sera had a definite cytotoxic effect on macrophages in vitro. This effect was observed when complement was added immediately or 4 hours after incubation with serum. It was lost after 24 hours. An inhibitory effects of AMS on phagocytosis in vivo has also been observed, which disappeared after one day. Antibody production in vivo was only moderately affected by a pretreatment with AMS. However, addition of AMS to spleen cell cultures stimulated with sheep red blood cells inhibited the appearance of hemolytic plaque forming cells.


Direct asymmetric organocatalytic Michael reactions of alpha,alpha-disubstituted aldehydes with beta-nitrostyrenes for the synthesis of quaternary carbon-containing products.
[reaction: see text] Direct asymmetric catalytic Michael reactions have been performed using chiral-amine/acid bifunctional catalysts. Performed with 0.3 equiv of (S)-(+)-1-(2-pyrrolidinylmethyl)pyrrolidine and 0.3 equiv of trifluoroacetic acid as the catalyst, the reaction of alpha,alpha-dialkylaldehydes with (E)-beta-nitrostyrene provided the alpha,alpha-dialkyl Michael products in up to 96% yield with up to 91% ee. With respect to enantioselectivity, l-proline was a poor catalyst of this class of Michael reactions.


Effects of L-lysine monohydrochloride on insulin and blood glucose levels in spinal cord injured rats.
Hyperglycemia in brain and spinal cord could aggravate neurologic impairment. Recent studies showed that L-lysine monohydrochloride (LMH) could increase the insulin secretion and regulate the blood glucose level. The aim of the present study was to investigate the effects of LMH on pancreatic islet B cells, the levels of endogenous insulin and blood glucose in spinal cord injured rats. Forty male Wistar rats were divided into four groups, namely, normal control group, model group, high-dose LMH group (621.5 mg/kg equal to LMH 1/8 LD50), and low-dose LMH group (310.8 mg/kg equal to LMH 1/16 LD50). The model of spinal cord injured rat was established by hemi-transection at the lower right thoracic spinal cord. LMH was administered via intraperitoneal injection once spinal cord injury was produced in rats. All rats were sacrificed 48 hours after spinal cord injured. The effects of LMH on pancreatic islet B cells, the content of endogenous insulin, and the level of blood glucose were observed with immunohistochemical method, radioimmunoassay method, and biochemical analyzer, respectively. The insulin immunohistochemical intensities of islet B cells were significantly weaker in model group than those in normal control group (P < 0.01). The levels of endogenous insulin were significantly lower and the blood glucose levels were significantly higher in model group than those in normal control group (P < 0.01). The insulin immunohistochemical intensities of islet B cells were significantly stronger in high-dose LMH group than those in model group (P < 0.05). In addition, we found that the levels of endogenous insulin were significantly higher and the blood glucose levels were significantly lower in high-dose LMH group than those in model group (P < 0.05). There were no significant differences in the insulin immunohistochemical intensities of islet B cells, the levels of endogenous insulin and the blood glucose between low-dose LMH group and model group (P > 0.05). LMH, but dose-dependent, might participate in the regulation of pancreatic islet B cells, and then reduce the blood glucose levels in the spinal cord injured rats.


Evaluation and prevention of enamel surface damage during dental restorative procedures.
This study examined the effect of operator experience, dominance, tooth position and access, on frequency and extent of iatrogenic damage to approximal tooth surfaces during conventional Class II cavity preparations. The effectiveness of protective devices in minimizing damage was also explored. 10 students and 10 experienced dentists each prepared 24 Class II cavity preparations in typodont teeth without protection; 10 utilizing stainless steel matrix bands and 10 utilizing protective wedges. The frequency and extent of damage were analysed with respect to the above variables. Subsequently, 20 natural and 20 typodont teeth were utilized to establish the relationship in depth of damage caused by a high-speed diamond bur on typodont versus natural teeth. Dentists caused iatrogenic damage on 74% of approximal surfaces without protection, which fell to 50% and 46% respectively when matrix bands and wedges were used as guards. The corresponding rates of damage for students were 94%, 80% and 44%. There was no difference in depth of damage caused on the two types of teeth when bur was in contact with teeth for a very short time. Greater operator experience and the use of guards reduces iatrogenic damage to proximal surfaces during preparation with high-speed rotary instruments.


Normal fetal thyroid volume.
A prospective study of 289 normal fetal thyroids was undertaken between 20 and 36 weeks of gestation in 289 mothers without any history of thyroid disease or maternal disease that could cause intrauterine growth restriction. The fetal thyroid volume was calculated by the ellipsoid equation, with the assumption that each lobe was an ellipse. A nomogram relating fetal thyroid volume to gestational age was established. The mean fetal thyroid volume was found to be 0.23 +/- 0.18 cm3 (median 0.17 cm3; range 0.022-0.85 cm3). The ratio of fetal thyroid volume to estimated fetal weight (V/W ratio) (at 0.163 +/- 0.079 cm3/kg) was constant throughout gestation. The value was smaller than those of the child and the adult reported in other studies, indicating that fetal thyroid enlargement is likely to be underestimated if based on postnatal criteria. It was found that the mean fetal thyroid volume was significantly correlated with estimated fetal weight and gestational age, in which the former dominated the effect. The fetal thyroid increased in size much faster after 32 weeks, suggesting that evaluation of possible fetal goiter should take into account gestational age and the nomogram developed in this study.


Recurrent case of pregnancy-induced atypical haemolytic uremic syndrome (P-aHUS).
Pregnancy-induced atypical haemolytic uremic syndrome (P-aHUS) is a rare condition characterised by microangiopathic haemolytic anaemia, thrombocytopenia and renal failure. It accounts for approximately 7% of total HUS cases. Here, we present a case of recurrent P-aHUS in a 25-year-old Hispanic woman. Pregnancy was the clear trigger in both instances, and the disease manifested in first week of the postpartum period. Because of her significant obstetric history, a multidisciplinary approach was adopted to monitor her second pregnancy antepartum and post partum. As the patient developed recurrence of P-aHUS 4 days after her delivery, she was immediately administered eculizumab within few hours of disease manifestation. The patient normalised her haematological parameters within 1 week but sustained dialysis-requiring renal failure for a total of 6 weeks. This case highlights the advances as well as the ongoing uncertainties, especially with respect to the use of eculizumab, in this rare but morbid disease.


Clinicopathological Features of Cases with Primary Breast Cancer not Identified by 18F-FDG-PET.
Several studies have reported that high F18-fluorodeoxyglucose (FDG) uptake is predictive of poor prognosis and aggressive features in patients with breast cancer. While these studies evaluated the prognostic value for cases with high FDG uptake, they did not elucidate the meaning of FDG negativity in primary breast cancer. In this study, we evaluated the clinicopathological features of breast cancer cases without FDG uptake. We retrospectively investigated the cases of 219 consecutive patients with primary breast cancer who underwent FDG-positron emission tomography (PET) preoperatively. Among the 219 patients, 25 (11.4%) did not have FDG uptake in the tumor. The 219 cases with breast cancer were divided into two groups based on the presence of FDG uptake in the primary tumor. The present univariate analysis revealed that histology, small invasive tumor size, high estrogen receptor (ER) or progesterone receptor (PgR) expression, low nuclear grade and absence of lymph node metastasis were significantly associated with negative FDG uptake in the primary tumor. On the other hand, the size of ductal spread was not significantly different between the two groups. Multivariate analysis revealed that small-size tumor invasion and lower nuclear grade were statistically significant. Among the 25 cases without FDG uptake, there was no recurrent disease in spite of there being no case that underwent chemotherapy, while 4 cases among the 194 cases with FDG uptake had disease recurrence. Our findings imply that preoperative FDG negativity in primary breast cancer is effective in predicting better prognosis, but is less effective in predicting ductal spread. Cases without FDG uptake in the primary tumor may have a lower risk of recurrent disease and may be able to safely avoid adjuvant chemotherapy.


Impacts of separate rejection water treatment on the overall plant performance.
A separate rejection water treatment appears as a high-tech unit process which might be recommendable only for specific cases of an upgrading of an existing wastewater treatment plant. It is not the issue of this paper to consider a specific separate treatment process itself but to investigate the influence of such a process on the overall plant performance. A plant-wide model has been applied as an innovative tool to evaluate effects of the implemented sidestream strategy on the mainstream treatment. The model has been developed in the SIMBA environment and combines acknowledged mathematical descriptions of the activated sludge process (ASM1) and the anaerobic mesophilic digestion (Siegrist model). The model's calibration and validation was based on data from 5 years of operating experience of a full-scale rejection water treatment. The impact on the total N-elimination efficiency is demonstrated by detailed nitrogen mass flow schemes including the interactions between the wastewater and the sludge lane. Additionally limiting conditions due to dynamic N-return loads are displayed by the model's state variables.


DNA-analysis is mandatory in case of an uncommon malignancy.
In a child a diagnosis of sweat gland carcinoma was made on basis of a surgical specimen presumed to be taken from an occipital lymph node. DNA analysis confirmed mixing of specimens in the referring hospital.


Core-shell vs. multi-shell formation in nanoalloy evolution from disordered configurations.
The evolution towards equilibrium of AuCo, AgNi and AgCu nanoparticles is studied by molecular dynamics simulations. Nanoparticle sizes of about 2.5 nm are considered, in the temperature range from 300 to 700 K. The simulations reveal complex equilibration pathways, in which geometric structure and chemical ordering change with time. These nanoparticles present the same type of strong tendency to phase separation and to surface segregation of either Au or Ag, which lead to the same type of core@shell equilibrium structures. In spite of these similarities, the equilibration pathways of these nanoparticles from chemically disordered configurations present both quantitative and qualitative differences. Quantitative differences are found in the equilibration time scale, which is much longer in AgCu than in AgNi and AuCo. Qualitative differences are found in the presence or absence of geometric structure transformations, and in the formation of different types of three-shell metastable chemical ordering during evolution. It is also shown that surface segregation depends on the geometric structure, being faster in icosahedra than in fcc nanoparticles.


Professional responsibility: implications for nursing practice within the realms of cardiothoracics.
Through an exploration of the realms of professional responsibility this paper aims to illustrate its implications in nursing practice with particular reference to the area of cardiothoracics. Throughout the paper issues surrounding the 'Code of Professional Conduct for the Nurse, Midwife and Health Visitor' and the 'The Scope of Professional Practice' released by the United Kingdom Central Council for Nursing, Midwifery and Health Visiting (U.K.C.C.) in 1992 are considered. 'The Code' as it will subsequently be referred to, and the 'The Scope of Professional Practice' (U.K.C.C.) address issues of current change within the nursing profession and provide a focus for discussion. While recognising both current constraints to clinical practice and the complexity of professional responsibility, it is concluded that the delivery of research based, quality patient care remains central to the role of all nursing professionals. With a primary focus on others before the self, nursing professionals must continue to address issues surrounding the maintenance and development of this care.


"HPV score", a scoring system for histological diagnosis of human papillomavirus infection in dysplasia of the uterine cervix.
We devised a scoring system, "HPV score", for histological diagnosis of human papillomavirus (HPV) infection in dysplasia of the uterine cervix. Four hundred and sixty cases of cervical dysplasia were screened for the presence of HPV infection using this system, and 116 cases (25%) were judged to be HPV infection. The results showed good correlation with those obtained using the immunoperoxidase (IMPO) method, but 42 of the 116 cases (36%) were negative for HPV antigen by the IMPO method. Severe stromal inflammation was noted in about half of such cases. Among the four histological types of cervical condyloma, the flat type was most common among the cases with HPV infection judged according to HPV score. It was concluded that this scoring system is simple and accurate for the detection of HPV infection in cervical lesions.


Microwave spectra and structure of the cyclopropanecarboxylic acid-formic acid dimer.
The rotational spectrum of the cyclopropanecarboxylic acid-formic acid doubly hydrogen bonded dimer has been measured in the 4-11 GHz region using a Flygare-Balle type pulsed-beam Fourier transform microwave spectrometer. Rotational transitions were measured for the parent, four unique singly substituted (13)C isotopologues, and a singly deuterated isotopologue. Splittings due to a possible concerted double proton tunneling motion were not observed. Rotational constants (A, B, and C) and centrifugal distortion constants (DJ and DJK) were determined from the measured transitions for the dimer. The values of the rotational (in MHz) and centrifugal distortion constants (in kHz) for the parent isotopologue are A = 4045.4193(16), B = 740.583 80(14), C = 658.567 60(23), DJ = 0.0499(16), and DJK = 0.108(14). A partial gas phase structure of the dimer was derived from the rotational constants of the measured isotopologues, previous structural work on each monomer units and results of the calculations.


Bile duct strictures after adult liver transplantation: a role for biliary reconstructive surgery?
There is no accurate method to determine the functional significance of bile duct strictures after liver transplantation, and although biliary reconstructive surgery (Roux-en-Y hepaticojejunostomy, HJ) is the second-line treatment in patients with persistent allograft dysfunction following failed endoscopic therapy, there is no evidence to support this approach. Liver transplant recipients with allograft dysfunction and demonstrable bile duct strictures who had undergone hepaticojejunostomy were identified from a prospective database. Preoperative and follow-up clinical, biochemical, and radiological data were collected. Perioperative liver biopsies were evaluated prospectively by two histopathologists blinded to clinical information. The biopsies were scored according to presence and severity of biliary features, fibrosis, and coexisting diseases. The effects of preoperative factors on postoperative allograft function were analyzed using SPSS statistical software. After hepatico-jejunostomy, graft function returned to normal in 8/44 patients (18%), improved in 16/44 (36%), but remained abnormal in 20/44 (45%), including 4 patients who subsequently underwent retransplantation. Hepaticojejunostomy was more likely to yield a favorable outcome (improved or normal graft function) when performed within 2 years of transplantation. Prolonged duration of biliary obstruction was associated with development of advanced graft fibrosis at the time of surgery, but neither factor significantly influenced postoperative graft function. In conclusion, biliary reconstruction successfully restores graft function in the majority of patients who present with anastomotic strictures within the first 2 years after liver transplantation. In patients presenting with bile duct strictures late after transplantation, surgery should be reserved for selected patients without histological evidence of graft fibrosis (moderate-severe) or significant nonbiliary pathology.


Electrostatic potential energy within a protein monitored by metal charge-dependent hydrogen exchange.
Hydrogen exchange measurements on Zn(II)-, Ga(III)-, and Ge(IV)-substituted Pyrococcus furiosus rubredoxin demonstrate that the log ratio of the base-catalyzed rate constants (Delta log k(ex)) varies inversely with the distance out to at least 12 A from the metal. This pattern is consistent with the variation of the amide nitrogen pK values with the metal charge-dependent changes in the electrostatic potential. Fifteen monitored amides lie within this range, providing an opportunity to assess the strength of electrostatic interactions simultaneously at numerous positions within the structure. Poisson-Boltzmann calculations predict an optimal effective internal dielectric constant of 6. The largest deviations between the experimentally estimated and the predicted DeltapK values appear to result from the conformationally mobile charged side chains of Lys-7 and Glu-48 and from differential shielding of the peptide units arising from their orientation relative to the metal site.


The role of ACE2 in the renin-angiotensin-system: Etiology and therapy of COVID-19 from a pharmaceutical perspective.
Angiotensin-2 converting enzyme (ACE2), a key element of the renin-angiotensin-system (RAS), is not only the direct target of infection by the human SARS-Cov-2 virus but is at the same the root for the complex pathogenetic events of COVID-19. From a pharmaceutical perspective, several established classes of medicines are involved in different phases of the disease. From their known mechanisms of action, a comprehensive understanding of COVID-19 will be hopefully soon delineated. A set of proven medicines is available to cope at least with some of the pathologies involved. To arrive back to normal life, vaccinations and broad consideration of hygienic measures are to be complemented by effective medicines to treat airborne viral infections. Therapeutic schemes based on a comprehensive understanding of the disease will include drug combinations made up from both established drugs as well as novel drugs presently under development.


Decabromodiphenyl ethane exposure damaged the asymmetric division of mouse oocytes by inhibiting the inactivation of cyclin-dependent kinase 1.
Decabromodiphenyl ethane (DBDPE) is a new brominated flame retardant and is widely added to flammable materials to prevent fire. Because it has been continuously detected in a variety of organisms and humans, it is important to reveal the biological toxicity of DBDPE. However, the influence of DBDPE for female reproduction is unclear. In this study, we investigated whether and how DBDPE exposure affects oocyte development. Female mice as a model were orally exposed to DBDPE by 0, 0.05, 0.5, 5, 50 μg/kg bw/day for 30 days (0.05 μg/kg bw/day is close to the environmental exposure concentration). We found that exposure of mice to DBDPE did not affect the first polar body extrusion (PBE) of oocytes. Strikingly, however, asymmetric division of oocytes was markedly impaired in 5 and 50 μg/kg bw/day DBDPE exposed group, which resulted in oocytes with larger polar bodies (PBs). Then, we further explored and found that DBDPE exposure inhibited the spindle migration and membrane protrusion in oocytes during anaphase of meiosis I (anaphase I), thereby impairing asymmetric division. Additionally, we found that DBDPE exposure suppressed the inactivation of cyclin-dependent kinase 1 (Cdk1), resulting in the decrease of cytoplasmic formin2 (FMN2)-mediated F-actin polymerization in oocytes at the onset of anaphase I. Simultaneously, DBDPE exposure damaged the structural integrity of the spindle and the perpendicular relationship between spindle and cortex. These together led to the failure of spindle migration and membrane protrusion required for oocytes asymmetric division. Finally, DBDPE exposure injured the development of blastocysts, leading to blastocyst apoptosis.


Long-chain n-3 polyunsaturated fatty acids dissociate phosphorylation of Akt from phosphatidylinositol 3'-kinase activity in rats.
We examined whether a low amount of dietary long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) modulated phosphatidylinositol 3'-kinase (PI 3-kinase) activity and downstream Akt phosphorylation differently in normal or insulin-resistant rats. Rats were fed for 28 days with either a control diet containing 14.6% of metabolizable energy (ME) as peanut-rape oil (PR) or an n-3 diet where 4.9% of ME as PR was replaced by fish oil. Over the last 5 days, rats received 9 per thousand NaCl or dexamethasone (1 mg/kg). Insulin stimulation of both PI 3-kinase activity and Akt serine(473) phosphorylation and modulation of GLUT4 content were studied in liver, muscle, and adipose tissue (AT). Glucose tolerance and insulin sensitivity were determined by an oral glucose challenge. In muscle and AT, LC n-3 PUFA abolished insulin-stimulated PI 3-kinase activity. These effects were not paralleled by defects in Akt serine(473) phosphorylation, which was even increased in AT. Dexamethasone abolished insulin-stimulated PI 3-kinase activity in all tissues, whereas Akt serine(473) phosphorylation was markedly reduced in muscle but unaltered in liver and AT. Such tissue-specific dissociating effects of LC n-3 PUFA on PI 3-kinase/Akt activation took place without alteration of glucose metabolism. Maintenance of a normal glucose metabolism by the n-3 diet despite abolition of PI 3-kinase activation was likely explained by a compensatory downstream Akt serine(473) phosphorylation. The inability of LC n-3 PUFA to prevent insulin resistance by dexamethasone could result from the lack of such a dissociation.


Importance of RNF213 polymorphism on clinical features and long-term outcome in moyamoya disease.
OBJECT Moyamoya disease (MMD) is an idiopathic cerebrovascular occlusive disorder prevalent in East Asia. In the pathogenesis of MMD, the important role of genetic factors is being elucidated, and RNF213 has recently been identified as a susceptibility gene for MMD. The aim of this retrospective study was to investigate the RNF213 genotype in patients with MMD and to determine their genotype-phenotype associations. METHODS The study involved 165 Korean MMD patients from 155 unrelated families who were diagnosed with MMD at a single center from 1995 to 2013. Their demographic, radiological, and clinical findings were evaluated. Direct sequencing of the major RNF213 single nucleotide polymorphisms was performed. The association of the common RNF213 variant with MMD risk was evaluated using historical controls for comparison. Correlations between RNF213 genotype and phenotype were statistically analyzed. RESULTS The c.14429G>A (p.R4810K) variant was identified in 125 (75.8%) of 165 MMD patients. Most patients (112) were heterozygous, and 13 patients had 2 copies of the c.14429G>A variant. A novel heterozygous variant, c.12086A>G (p.Q4029R), was found in 1 additional patient. The minor allele frequency of the c.14429G>A variant was significantly higher in the MMD group (138 [41.8%] of 330 patients) than in the control group (8 [1.36%] of 588 subjects; p < 0.001). The c.14429G>A (p.R4810K) variant significantly increased the risk of MMD in Korean patients, with an OR of 52.11 (p < 0.001) compared with controls. Moreover, c.14429G>A (p.R4810K) genotypes occurred more frequently in patients with a family history of MMD. The homozygous variant was highly associated with early-onset MMD (age at onset < 5 years), cerebral infarction at diagnosis, and cognitive impairment in long-term outcome. CONCLUSIONS The findings indicate that the c.14429G>A (p.R4810K) allele of RNF213 is strongly associated with Korean patients with MMD. The homozygous c.14429G>A (p.R4810K) variant is particularly related to early-onset MMD, severe symptomatic manifestations at diagnosis, and poor prognosis. This genotypic variant may be a useful biomarker for early-onset MMD or unstable MMD with cerebral infarction, which requires early diagnosis and revascularization treatment.


[Efficacy of Activated Autologous Hematopoietic Stem Cell Transplantation in the Treatment of Acute Myeloid Leukemia].
To compare the efficacy of activated autologous bone marrow and peripheral blood hematopoietic stem cell transplantation (Auto-HSCT) and matched sibling donor allogeneic hematopoietic stem cell transplantation (MSD-HSCT) for the first complete remission of adult acute myeloid leukemia (AML-CR1). For 86 adult patients with first complete remission of AML who underwent auto-HSCT (41 cases) and MSD-HSCT (45 cases) in our hospital from June 2012 to June 2020, the patients were treated with modified MAC [Malflane 160 mg/(m2·d), -3 days, Ara-C 2 g/(m2·2), -3 days 21∶00, -2 days 9∶00, CTX 60 mg/(kg·d),-3 d, -2 d], the stem cells were activated by IL-2 (1 000 U/ mL), IFN-α (100 U/ mL) and IFN-γ (100 U/ml). The overall survival (OS), leukemia free survival (LFS), cumulative incidence of recurrence (CIR) and non-recurrence mortality (NRM) of patients with different types of transplantation were compared. The 3-year OS rates of Auto-HSCT group and MSD-HSCT group were 75% and 69.5%, and the 3-year LFS rates were 70.6% and 82.4%, respectively. There was no statisticaly significant difference in the 3-year OS rates of low risk, medium risk and high risk patients in the Auto-HSCT and MSD-HSCT group (90.2% vs 87.5%, 68.4% vs 68.8%, 28.6% vs 53.3%), the LFS rates of low risk, medium risk and high risk patients in the auto-HSCT and MSD-HSCT group were 90.2% and 87.5%(P=0.838), 71.8% and 91.7%(P=0.184), 0 and 67.5%(P=0.027), respectively. The NRM of Auto-HSCT and MSD-HSCT group were 4.9% and 20% (P=0.036), and CIR were 24.4% and 13.3% (P=0.188). Univariate analysis showed that the survival time of patients was significantly correlated with the number of CR courses and disease risk stratification (P=0.005, P=0.000). Cox multivariate analysis showed that disease risk stratification was an independent risk factor affecting OS (P=0.001). For adult patients with primary AML-CR1, Auto-HSCT is safe and effective. In the absence of sibling donor, Auto-HSCT can be regarded as an effective post-remission treatment for patients with intermediate risk AML-CR1.


Five Typical Stenches Detection Using an Electronic Nose.
This paper deals with the classification of stenches, which can stimulate olfactory organs to discomfort people and pollute the environment. In China, the triangle odor bag method, which only depends on the state of the panelist, is widely used in determining odor concentration. In this paper, we propose a stenches detection system composed of an electronic nose and machine learning algorithms to discriminate five typical stenches. These five chemicals producing stenches are 2-phenylethyl alcohol, isovaleric acid, methylcyclopentanone, γ-undecalactone, and 2-methylindole. We will use random forest, support vector machines, backpropagation neural network, principal components analysis (PCA), and linear discriminant analysis (LDA) in this paper. The result shows that LDA (support vector machine (SVM)) has better performance in detecting the stenches considered in this paper.


Majorana Braiding with Thermal Noise.
We investigate the self-correcting properties of a network of Majorana wires, in the form of a trijunction, in contact with a parity-preserving thermal environment. As opposed to the case where Majorana bound states are immobile, braiding Majorana bound states within a trijunction introduces dangerous error processes that we identify. Such errors prevent the lifetime of the memory from increasing with the size of the system. We confirm our predictions with Monte Carlo simulations. Our findings put a restriction on the degree of self-correction of this specific quantum computing architecture.


Myelodysplastic Syndrome with t(1;14),t(1;17),t(1;19) Transforms to AML-M5: A Case Report and Literature Review.
Chromosomal aberrations play an important role in the incidence of myelodysplastic syndromes (MDS) and development to acute myeloid leukemia (AML). We report a case of a 62-year-old male patient diagnosed with MDS with excess blasts. The karyotype was 45, XY,+1,+1,-7,-10,-22,t(1;14) (q21;q32),t(1;17)(q21;p13),t(1;19)(q21;p13). The patient and his family refused treatment for financial reasons. After 2 months, the patient's MDS transformed into acute myeloid monocytic leukemia (AML-M5). This case of MDS with poor prognosis shows that patients with chromosomal numerical abnormality and balanced translocations should be treated early to prevent transition to AML. Further study of this case will reveal the molecular mechanism of MDS-to-AML transformation and identify new leukemic fusion genes.


Automated determination of blood metabolic acidosis, total acid phosphatase, angiotensin converting enzyme, fructosamine and urinary oxalate and citrate on the ABX Pentra 400.
We have adapted automated assays for quantification of pyruvate (PYR), acetoacetate (AA), and betahydroxybutyrate (BOHB) in plasma, total acid phosphatase (TAP), angiotensin converting enzyme (ACE) and fructosamine (FRUCT) in serum, and oxalate (OXAL) and citrate (CIT) in urine that can be performed on the ABX Pentra 400. The aim of this study was to evaluate the analytical performances of these parameters and to compare data obtained in patient samples with those obtained with the Cobas Mira. Within-run and between-run imprecisions ranged from 0.55 to 14.2 and 0.37 to 11.8%, respectively. Correlations with the Cobas Mira showed r2 coefficients ranging from 0.816 to 0.983 and linear regression slopes from 0.706 to 1.087. The ABX Pentra 400 provides precise and accurate measurement for a wide variety of analytes including PYR, AA, BOHB, TAP, ACE, FRUCT in blood samples and OXAL, CIT in urine.


Escherichia coli O157:H7 survival, biofilm formation and acid tolerance under simulated slaughter plant moist and dry conditions.
Microorganisms persisting in slaughter plant environments may develop acid resistance and be translocated to other environmental surfaces or products. The objective of this study was to evaluate the potential of Escherichia coli O157:H7 to form biofilms and maintain acid resistance, under different culture habituation scenarios, on stainless steel coupons (2 x 5 x 0.08 cm), in the presence of beef carcass decontamination runoff fluids (washings). Coupons were stored in test tubes with unsterilized water washings (WW; pH 6.94) or lactic acid washings (LAW; pH 4.98), which were inoculated with E. coli O157:H7 (10(3)-10(4)CFU/ml) and incubated at 15 (24 or 48 h) or 35 degrees C (7 or 24 h), simulating different habituation scenarios on sites of a slaughter plant, including sanitation and overnight drying, during consecutive operational shifts. Acid resistance (AR) of planktonic and detached E. coli O157:H7 cells was assessed in tryptic soy broth adjusted to pH 3.5 with lactic acid. The highest pre-drying attachment and AR of E. coli O157:H7 were observed after 24h at 35 degrees C and 48 h at 15 degrees C. Drying reduced (P<0.05) recovery of attached E. coli O157:H7 cells; however, exposure of dried coupons to uninoculated washings allowed recovery of attached E. coli O157:H7, which restored AR, especially under conditions that favored post-drying growth. Exposure of attached cells to 50 ppm PAA for 45 s before drying, as well as habituation in LAW, reduced the recovery and AR of E. coli O157:H7. Therefore, incomplete removal of biofilms may result in cells of increased AR, especially in sites within a slaughter plant, in which liquid meat wastes may remain for long periods of time.


A novel formulation for mebeverine hydrochloride.
The antispasmodic drug mebeverine hydrochloride was formulated into a film-forming gel to be used as a topical local anesthetic. A mixture of cellulose derivatives was used as a base. Additives were used to enhance the release as well as the residence time. Formulations were characterized in terms of drug release, mucoadhesion and rheology. Clinically, the selected formula has shown faster onset (p = 0.0156), longer duration (p = 0.0313), better film residence (p = 0.0313), and no foreign body sensation (p = 0.0313) in comparison to Solcoseryl dental paste. Histopathological examination showed no change in inflammatory cells count, concluding that this topical anesthetic is efficacious and safe orally.


Bubbles in solvent microextraction: the influence of intentionally introduced bubbles on extraction efficiency.
Significant improvements to microdrop extractions of triazine pesticides are realized by the intentional incorporation of an air bubble into the solvent microdroplet used in this microextraction technique. The increase is attributed partly to greater droplet surface area resulting from the air bubble being incorporated into the solvent droplet as opposed to it sitting thereon and partly to thin film phenomena. The method is useful at nanogram/liter levels (LOD 0.002-0.012 μg/L, LOQ 0.007-0.039 μg/L), is precise (7-12% at 10 μg/L concentration level), and is validated against certified reference materials containing 0.5 and 5.0 μg/L analyte. It tolerates water and fruit juice as matrixes without serious matrix effects. This new development brings a simple, inexpensive, and efficient preconcentration technique to bear which rivals solid phase microextraction methods.


Extensile lateral versus sinus tarsi approach for calcaneal fractures: A meta-analysis.
Calcaneal fractures are the most common tarsal bone fracture, and are often accompanied by heel pain, local swelling, tenderness, and inability to walk or stand. Surgical intervention results in better reconstruction of the calcaneal anatomy and reduces future complications; however, the optimal incision approach is still controversial. The incision is exposed better with extensile lateral approach (ELA), while the sinus tarsi approach (STA) causes fewer complications. The purpose of this meta-analysis is to compare the outcomes of STA and ELA. Published trials comparing ELA and STA in calcaneal fractures were included in our analysis. The quality of each study was assessed using the revised Jadad scale and the Newcastle-Ottawa scale. Two researchers (CP and BY) independently extracted data from all selected studies. Fixed- or random-effects models with mean differences and odds ratios were used to pool the continuous and dichotomous variables to determine the heterogeneity of the included studies. Calcaneal height and calcaneal width had high heterogeneity. Results showed that the incidence of incision complications in STA was lower than that in ELA (P < .001). There was high heterogeneity in operative time (I2 = 97%), length of hospital stay (I2 = 98%), Böhler angle (I2 = 80%), Gissane angle (I2 = 98%), and American Orthopaedic Foot & Ankle Society scores (I2 = 73%). No source of heterogeneity was found by sensitivity analysis, subgroup analysis, or regression analysis, and the random-effects model was used. STA operative time was significantly shorter than ELA (P < .001). Length of hospital stay after STA was significantly shorter than after ELA (P = .002). There was no statistical difference in the Böhler and Gissane angles between STA and ELA. Postoperative American Orthopaedic Foot & Ankle Society scores after STA were higher than after ELA (P = .01). Results show that, compared with ELA, STA is superior for treating calcaneal fractures due to anatomical reduction of the calcaneus, reduction of incision complications incidence, and shortened operative time and postoperative stay.


Nitration of TRPM2 as a Molecular Switch Induces Autophagy During Brain Pericyte Injury.
Dysfunction of neurovascular pericytes underlies breakdown of the blood-brain barrier, but the molecular mechanisms are largely unknown. In this study, we evaluated the role of the transient receptor potential melastatin-related 2 (TRPM2) channel and autophagy during brain pericyte injury both in vitro and in vivo. A rapid induction in autophagy in human brain vascular pericytes, in the zinc oxide nanoparticles (ZnO-NP)-induced cell stress model, was paralleled with an increase in the expression of the TRPM2-S truncated isoform, which was abolished by treatment with a nitric oxide synthase inhibitor and a peroxynitrite scavenger. Furthermore, Y1485 in the C-terminus of the TRPM2 protein was identified as the tyrosine nitration substrate by mass spectrometry. Overexpression of the Y1485S TRPM2 mutant reduced LC3-II accumulation and pericyte injury induced by ZnO-NP. Consistently, LC3-II accumulation was reduced and pericytes were better preserved in intact brain microvessels of the TRPM2 knockout mice after ZnO-NP-induced vascular injury. Innovation and Conclusions: Our present study has revealed a novel mechanism of autophagy disturbance secondary to nitrosative stress-induced tyrosine nitration of TRPM2 during pericyte injury. Antioxid. Redox Signal. 27, 1297-1316.


The inventory as a core element in the further development of the science curriculum in the Mannheim Reformed Curriculum of Medicine.
Introduction: The German Council of Science and Humanities as well as a number of medical professional associations support the strengthening of scientific competences by developing longitudinal curricula for teaching scientific competences in the undergraduate medical education. The National Competence Based Catalogue of Learning Objectives for Undergraduate Medical Education (NKLM) has also defined medical scientific skills as learning objectives in addition to the role of the scholar. The development of the Mannheim science curriculum started with a systematic inventory of the teaching of scientific competences in the Mannheim Reformed Curriculum of Medicine (MaReCuM). Methods: The inventory is based on the analysis of module profiles, teaching materials, surveys among experts, and verbatims from memory. Furthermore, science learning objectives were defined and prioritized, thus enabling the contents of the various courses to be assigned to the top three learning objectives. Results: The learning objectives systematic collection of information regarding the current state of research, critical assessment of scientific information and data sources, as well as presentation and discussion of the results of scientific studies are facilitated by various teaching courses from the first to the fifth year of undergraduate training. The review reveals a longitudinal science curriculum that has emerged implicitly. Future efforts must aim at eliminating redundancies and closing gaps; in addition, courses must be more closely aligned with each other, regarding both their contents and their timing, by means of a central coordination unit. Conclusion: The teaching of scientific thinking and working is a central component in the MaReCuM. The inventory and prioritization of science learning objectives form the basis for a structured ongoing development of the curriculum. An essential aspect here is the establishment of a central project team responsible for the planning, coordination, and review of these measures.


A method for parametric estimation of the number and size distribution of cell clusters from observations in a section plane.
The problem of finding the number and size distribution of cell clusters that grow in an organ or tissue from observations of the number and sizes of transections of such cell clusters in a planar section is considered. This problem is closely related to the well-known corpuscle or Wicksell problem in stereology, which deals with transections of spherical objects. However, for most biological applications, it is unrealistic to assume that cell clusters have spherical shapes since they may grow in various ways. We therefore propose a method that allows for more general spatial configurations of the clusters. Under the assumption that a parametric growth model is available for the number and sizes of the cell clusters, expressions are obtained for the probability distributions of the number and sizes of transections of the clusters in a section plane for each point in time. These expressions contain coefficients that are independent of the parametric growth model and time but depend on which model is chosen for the configuration of the cell clusters in space. These results enable us to perform estimation of the parameters of the growth model by maximum likelihood directly on the data instead of having to deal with the inverse problem of estimation of three-dimensional quantities based on two-dimensional data. For realistic choices of the configuration model, it will not be possible to obtain the exact values of the coefficients, but they can easily be approximated by means of computer simulations of the spatial configuration. Monte Carlo simulations were performed to approximate the coefficients for two particular spatial configuration models. For these two configuration models, the proposed method is applied to data on preneoplastic minifoci in rat liver under the assumption of a two-event model of carcinogenesis as the parametric growth model.


Role of endogenous PACAP in catecholamine secretion from the rat adrenal gland.
We elucidated the contribution of endogenous pituitary adenylate cyclase-activating polypeptide (PACAP) to neurally evoked catecholamine secretion from the isolated perfused rat adrenal gland. Infusion of PACAP (100 nM) increased adrenal epinephrine and norepinephrine output. The PACAP-induced catecholamine output responses were inhibited by the PACAP type I receptor antagonist PACAP- (6-38) (30-3,000 nM) but were resistant to the PACAP type II receptor antagonist [Lys1,Pro2,5,Ara3,4,Tyr6]-vasoactive intestinal peptide (LPAT-VIP; 30-3,000 nM). Transmural electrical stimulation (ES; 1-10 Hz) or infusion of ACh (6-200 nM) increased adrenal epinephrine and norepinephrine output. PACAP-(6-38) (3,000 nM), but not LPAT-VIP, also inhibited the ES-induced catecholamine output responses. However, PACAP-(6-38) did not affect the ACh-induced catecholamine output responses. PACAP at low concentrations (0.3-3 nM), which had no influence on catecholamine output, enhanced the ACh-induced catecholamine output responses, but not the ES-induced catecholamine output responses. These results suggest that PACAP is released from the nerve endings to facilitate the neurally evoked catecholamine secretion through PACAP type I receptors in the rat adrenal gland.


Shed membrane particles from preeclamptic women generate vascular wall inflammation and blunt vascular contractility.
We investigated the role of microparticles in vascular dysfunction of the multisystemic disorder of preeclampsia in women's omental arteries or mouse arteries. Preeclamptic women displayed increased circulating levels of leukocyte- and platelet-derived microparticles compared with healthy pregnant individuals. Microparticles from preeclamptic, but not healthy, pregnant women induced ex vivo vascular hyporeactivity to serotonin in human omental arteries and mouse aortas. Hyporeactivity was reversed by a nitric-oxide (NO) synthase inhibitor and associated with increased NO production. In the presence of a cyclooxygenase (COX)-2 inhibitor, serotonin-mediated contraction was partially reduced in arteries treated with healthy microparticles but was abolished after treatment with preeclamptic microparticles. This was associated with increased 8-isoprostane production. Preeclamptic microparticles induced up-regulation of inducible nitric-oxide synthase and COX-2 expression, evoked nuclear factor-kappaB activation, and enhanced oxidative and nitrosative stress. Interestingly, the microparticles originating most probably from leukocytes were responsible for the COX-2 vasoconstrictor component of preeclamptic microparticles, whereas those of platelet origin were mainly involved in NO release. Moreover, vascular hyporeactivity was observed in arteries taken from mice treated in vivo with preeclamptic microparticles. This study demonstrates pathophysiological relevance and provides a paradoxical effect of preeclamptic microparticles associated with proinflammatory properties on vessels, leading to enhanced NO and superoxide anion levels and counteraction of increased COX-2 metabolites.


Nonlinear optical properties and passively Q-switched laser application of a layered molybdenum carbide at 639 nm.
Molybdenum carbide (Mo2C) exhibits enormous potential applications in various optoelectronic and photonic fields due to its remarkably electrical and optical characteristics. Here, we fabricate a high-quality Mo2C film by the radio frequency magnetron sputtering deposition method. The nonlinear optical response and ultrafast dynamics are thoroughly studied based on open-aperture Z-scan and nondegenerate pump-probe experimental measurements. The open-aperture Z-scan experimental result exhibits a modulation depth of 8.5% and a saturation fluence of 0.28 mJ/cm2. Simultaneously, the relaxation time constant is fitted by a biexponential decay function, showing an ultrafast intraband carrier recovery time of 0.58 ps at 530 nm. Consequently, by employing the Mo2C film as a saturable absorber (SA), stable Q-switched Pr:YLF laser pulses with the shortest pulse width of 160 ns are generated at 639 nm. Our experimental results demonstrate excellent nonlinear optical properties of the layered Mo2C in the visible region and will further advance their potential applications in visible nonlinear optics.


Degradation of meta-trifluoromethylbenzoate by sequential microbial and photochemical treatments.
m- and p-trifluoromethyl (TFM)-benzoates are incompletely degraded by aerobic bacteria that catabolize alkylbenzoates; biodegradation ceases after ring-fission with the accumulation of a trifluoromethyl muconate semialdehyde (2-hydroxy-6-oxo-7,7,7-trifluorohepta-2,4-dienoate, TFHOD) which is resistant to biochemical attack. A bacterium (Strain V-1), isolated from sea-water, grew aerobically on benzoate or m-toluate. Cells grown on benzoate or m-toluate oxidized both compounds at similar relative rates. Catabolism involved benzoate 1,2-dioxygenase (decarboxylating) and meta-cleavage to yield muconate semialdehydes. Cells grown on benzoate metabolized m-TFM-benzoate to TFHOD. The ring-fission products from m-toluate and TFHOD were degraded by sunlight, and equimolar fluoride was released from TFHOD. Sequential biochemical and photochemical treatment allowed the destruction of m-TFM-benzoate beyond the biochemically recalcitrant intermediate TFHOD.


Understanding the Boron-Nitrogen Interaction and Its Possible Implications in Drug Design.
2-Aminoethoxydiphenylborate (2-APB) is a broad-spectrum modulator of various membrane proteins. Specifically, it exhibits concentration dependent modulation of calcium signaling through store-operated calcium (SOC) channels: low micromolar concentration of 2-APB stimulates SOC entry while a higher concentration induces complete inhibition. Ab initio quantum chemical calculations show that the relative stability of the two major isomers of 2-APB (cyclic and extended) is about 8 kcal/mol. The dual functionality of 2-APB for SOC channels is thus likely associated with its ability to switch among isomeric forms, suited to different binding sites in the SOC channels with distinct binding affinities. Importantly, the moderate relative stability of different isomers results from a delicate balance between the intramolecular boron-nitrogen coordinate bond with strength about -45 kcal/mol and ring strain engendered by cyclic oligomerization. The synergistic effect of these two factors likely makes 2-APB an ideal dual effect drug.


Evaluation of a Web-based malaria risk reduction game for study abroad students.
Compare feedback strategies in 3 versions of an educational game. Study abroad students (N = 482) participated by playing the game and completing pregame/postgame surveys January-March 2010. This study employed an experimental design. Primary outcome measures were knowledge gain, player satisfaction, and risk perception. One-third had previously traveled to a malaria-risk region, and two thirds planned to do so. Baseline malaria knowledge was low. Postgame knowledge and risk perception were significantly higher than pregame, irrespective of past travel status. The group that automatically received explanatory feedback following game decisions scored higher for mean knowledge gain, without differences in player satisfaction. The challenges of designing a feedback strategy to support Web-based learning make these results highly relevant to health educators developing interactive multimedia interventions. The increasing number of students traveling to higher-risk destinations demands attention. Both malaria-naive and malaria-experienced students would benefit from this approach to travel health education.


Roles of diversifying selection and coordinated evolution in the evolution of amphibian antimicrobial peptides.
Antimicrobial peptides are expressed in the skin of amphibians and are used to prevent infection by microorganisms. Frog species store distinct collections of antimicrobial peptides that show variation in size, charge, conformation, and bactericidal activity, and so the evolution of antimicrobial peptide gene families may reflect the adaptive diversification of these loci. We examined the molecular evolution of antimicrobial peptide transcripts from hylid and ranid frog species. Our results show that after the gene family arose in the common ancestor of the Hylidae and Ranidae, before the divergence of these families in the Mesozoic, it subsequently diversified within these groups with numerous duplication events and divergence of loci. Moreover, we provide evidence that suggests that members of the antimicrobial peptide gene family have been subject to diversifying selection within both propiece and mature domains of hylids and solely within the mature domain of ranids. Finally, our results suggest that coordinated and compensatory amino acid replacements have occurred within the acidic propiece and cationic mature domain of hylid antimicrobial peptide precursors, as has been observed for mammalian defensin genes, but not among those of ranid precursors.


Implication of a new molecule IK in CD34+ hematopoietic progenitor cell proliferation and differentiation.
HLA-DR is one of the markers associated with hematopoietic cell differentiation, since expression of this molecule is modulated throughout hematopoiesis. We have previously described and cloned the gene encoding factor IK, which inhibits both interferon gamma (IFN-gamma)-induced and constitutive HLA-DR expression. The current study demonstrates that IK gene transcripts are present in CD34+ cells purified from human umbilical cord blood. IK expression increased and was therefore inversely correlated with the gradual loss of HLA-DR during growth factor-induced CD34+ cell proliferation and differentiation. To study the possible role of IK in hematopoiesis, antisense probes were used. IK expression was specifically inhibited by an antisense oligodeoxynucleotide containing two phosphorothioate internucleotide linkages at each of the 3' and 5' ends and corresponding to the initiation site of IK mRNA. A control oligonucleotide was also tested in parallel. A specific decrease of IK transcripts was correlated with an increase of HLA-DR antigen expression level. In colony-forming assays, IK antisense oligonucleotide inhibited colony formation by multilineage early erythroid and granulomonocytic CD34+ progenitors. The mean colony size was decreased 70% by IK antisense oligonucleotide in comparison to controls. These results provide evidence that the IK molecule participates in the regulation of HLA-DR expression on hematopoietic cells and plays a role in growth factor-dependent CD34+ cell proliferation and differentiation by modulating HLA-DR expression.