Development of an electrophoretic method based on nanostructured materials for HbA1c determination.
Glycosylated hemoglobin (HbA1c) detection is performed routinely in hospitals as it is the most widespread confirmatory diagnosis of diabetes mellitus. Here we present a novel CE method for measuring HbA1c by introducing silica nanoparticles (NPs) modified with a boronic acid derivative (sugar loadings of 51 ± 2 μg/mg) as pseudo-stationary phase. Before the sample injection, SiO2 NP─B(OH)2 were introduced via pressure. Electrophoretic separation was explored through variation of the buffer pH and separation voltage, being the best separation, resolution and shorter separation time achieved with a 25 mM phosphate buffer pH 6.5. The calibration curve obtained was expressed as Area = 182.05%-1 × HbA1c - 377.02; R2 = 0.9826, using a UV/VIS absorbance detector at 415 nm (diode array). No interferences were observed from carbamylated or acetylated hemoglobin and the method shows a noteworthy stability. A paired t-test was applied to compare the developed CE method with a commercial HbA1c test and no significant variations have been observed at a 90% significance level.


Synergies between Fibrillated Nanocellulose and Hot-Pressing of Papers Obtained from High-Yield Pulp.
To extend the application of cost-effective high-yield pulps in packaging, strength and barrier properties are improved by advanced-strength additives or by hot-pressing. The aim of this study is to assess the synergic effects between the two approaches by using nanocellulose as a bulk additive, and by hot-pressing technology. Due to the synergic effect, dry strength increases by 118% while individual improvements are 31% by nanocellulose and 92% by hot-pressing. This effect is higher for mechanical fibrillated cellulose. After hot-pressing, all papers retain more than 22% of their dry strength. Hot-pressing greatly increases the paper's ability to withstand compressive forces applied in short periods of time by 84%, with a further 30% increase due to the synergic effect of the fibrillated nanocellulose. Hot-pressing and the fibrillated cellulose greatly decrease air permeability (80% and 68%, respectively) for refining pretreated samples, due to the increased fiber flexibility, which increase up to 90% using the combined effect. The tear index increases with the addition of nanocellulose, but this effect is lost after hot-pressing. In general, fibrillation degree has a small effect which means that low- cost nanocellulose could be used in hot-pressed papers, providing products with a good strength and barrier capacity.


Acute effects of moderate alcohol consumption on blood pressure and plasma catecholamines.
This study examines the response of blood pressure, plasma catecholamines and cortisol to acute alcohol intake in young men with light to moderate drinking habits. Ingestion of alcohol was associated with a highly significant increase in systolic blood pressure and heart rate which occurred before blood alcohol reached its peak concentration of 16.9 +/- 1.1 mmol/l (80 mg/100 ml). After an initial non-specific rise, diastolic pressure fell below values observed after drinking water only. This predominant effect of alcohol on systolic blood pressure is also seen with chronic alcohol consumption. Drinking water and non-alcoholic cold liquids caused a marked fall in plasma adrenaline and a transient rise in noradrenaline concentration. In contrast, drinking alcohol resulted in a relative rise in adrenaline and a delayed increase in noradrenaline concentration. Blood glucose increased after alcohol, supporting a physiological effect of adrenaline on liver glycogenolysis. Plasma cortisol concentration was also significantly higher after drinking alcohol. It is proposed that the relative rise in adrenaline together with higher cortisol levels, repeated over a variable period in susceptible individuals, are implicated in the elevation of blood pressure associated with long term alcohol consumption. It concurs with observations in man and experimental animals of a slow pressor mechanism mediated by adrenaline.


Topical Delivery of Tenofovir Disoproxil Fumarate and Emtricitabine from Pod-Intravaginal Rings Protects Macaques from Multiple SHIV Exposures.
Topical preexposure prophylaxis (PrEP) against HIV has been marginally successful in recent clinical trials with low adherence rates being a primary factor for failure. Controlled, sustained release of antiretroviral (ARV) drugs may help overcome these low adherence rates if the product is protective for extended periods of time. The oral combination of tenofovir disoproxil fumarate (TDF) and emtricitabine (FTC) is currently the only FDA-approved ARV drug for HIV PrEP. A novel pod-intravaginal ring (IVR) delivering TDF and FTC at independently controlled rates was evaluated for efficacy at preventing SHIV162p3 infection in a rigorous, repeat low-dose vaginal exposure model using normally cycling female pigtailed macaques. Six macaques received pod-IVRs containing TDF (65 mg) and FTC (68 mg) every two weeks, and weekly vaginal exposures to 50 TCID50 of SHIV162p3 began one week after the first pod-IVR insertion. All pod-IVR-treated macaques were fully protected throughout the study (P = 0.0002, Log-rank test), whereas all control animals became infected with a median of 4 exposures to infection. The topical, sustained release of TDF and FTC from the pod-IVR maintained protective drug levels in macaques over four months of virus exposures. This novel and versatile delivery system has the capacity to deliver and maintain protective levels of multiple drugs and the protection observed here warrants clinical evaluation of this pod-IVR design.


Exploring norms and beliefs related to AIDS prevention among California Hispanic men.
A focus-group study gathered qualitative information from Hispanic males in California regarding ethnic factors that might enhance or interfere with AIDS prevention efforts. A predominantly working-class convenience sample of 75 men aged 18-40 participated in seven groups of 10-12 participants each. Three groups were composed of single men, three of married men and one was composed of homosexual and bisexual men. Four groups included only participants born outside the United States; and Spanish was the preferred language in five of the groups. Evaluation of the discussions indicates that while AIDS awareness is high among this population, condom use is sporadic. Few participants knew that someone infected with the human immunodeficiency virus could appear healthy; most believed they could easily determine if a woman is risky by her appearance or social standing. Although many participants thought HIV could be transmitted through casual contact, the homosexual and bisexual men were more likely to acknowledge that they were at some risk of infection; most of the heterosexual men did not perceive themselves as being at risk, even those who engaged in sex with prostitutes or women they met at bars. The participants had ambivalent attitudes toward women who suggested condom use and were often reluctant to initiate condom use themselves. Religion did not seem to play a major role in the men's attitudes about AIDS, sexuality and condom use.


Tumor necrosis factor alpha modulates airway smooth muscle function via the autocrine action of interferon beta.
Current evidence suggests that tumor necrosis factor alpha (TNFalpha) and the family of interferons (IFNs) synergistically regulate many cellular responses that are believed to be critical in chronic inflammatory diseases, although the underlying mechanisms of such interaction are complex, cell-specific, and not completely understood. In this study, TNFalpha in a time-dependent manner activated both janus tyrosine kinase 1 and Tyk2 tyrosine kinase and increased the nuclear translocation of interferon-regulatory factor-1, STAT1, and STAT2 in human airway smooth muscle cells. In cells transfected with a luciferase reporter, TNFalpha stimulated gamma-activated site-dependent gene transcription in a time- and concentration-dependent manner. Using neutralizing antibodies to IFNbeta and TNFalpha receptor 1, we show that TNFalpha-induced secretion of IFNbeta mediated gamma-activated site-dependent gene expression via activation of TNFalpha receptor 1. In addition, neutralizing antibody to IFNbeta also completely abrogated the activation of interferon stimulation response element-dependent gene transcription induced by TNFalpha. Secreted IFNbeta acted as a negative regulator of TNFalpha-induced interleukin-6 expression, while IFNbeta augmented TNFalpha-induced RANTES (regulated on activation normal T cell expressed and secreted) secretion but had little effect on TNFalpha-induced intercellular adhesion molecule-1 expression. Furthermore TNFalpha, a modest airway smooth muscle mitogen, markedly induced DNA synthesis when cells were treated with neutralizing anti-IFNbeta. Together these data show that TNFalpha, via the autocrine action of IFNbeta, differentially regulates the expression of proinflammatory genes and DNA synthesis.


Purification of actin from fission yeast Schizosaccharomyces pombe and characterization of functional differences from muscle actin.
Fission yeast Schizosaccharomyces pombe is an important genetic model organism for studying the mechanisms of endocytosis and cytokinesis. However, most work on the biochemical properties of fission yeast actin-binding proteins has been done with skeletal muscle actin for matters of convenience. When simulations of mathematical models of the mechanism of endocytosis were compared with events in live cells, some of the reactions appeared to be much faster than observed in biochemical experiments with muscle actin. Here, we used gelsolin affinity chromatography to purify actin from fission yeast. S. pombe actin shares many properties with skeletal muscle actin but has higher intrinsic nucleotide exchange rate, faster trimer nucleus formation, faster phosphate dissociation rate from polymerized actin, and faster nucleation of actin filaments with Arp2/3 complex. These properties close the gap between the biochemistry and predictions made by mathematical models of endocytosis in S. pombe cells.


Ultrasound Features of Middle Cervical Sympathetic Ganglion.
To evaluate the feasibility of ultrasound (US) detection of the middle cervical sympathetic ganglion (CSG). We examined 52 consecutive patients with 104 bilateral middle CSGs. We identified middle CSG according to our sonographic assessment criteria. After finding oval-shaped hypoechoic structure on transverse scan, we verified a continuous, hypoechoic line connecting the bulbous structure at both sides. The basic US characteristics were evaluated using the size, location, relationship to the common carotid artery, and relationship to the inferior thyroidal artery. The middle CSG was identified in 41% (43/104). The mean size of the middle CSG was 3.8±1.5 mm in width, 1.9±0.7 mm in height, and 8.7±3.2 mm in length. They were primarily located at the C6 level (41/43, 95%). Regarding the relationship with common carotid artery, 88% (38/43) were lateral type and 12% (5/43) were medial type. Regarding the relationship with the inferior thyroid artery, 34% (15/43) were anterior type, 26% (11/43) were posterior type, and 40% (17/43) were unable to access the relationship. Direct visualization of middle CSG based on the understanding of both its location and US features may reduce complications and improve the accuracy of US-guided neck procedures.


TRAM is involved in IL-18 signaling and functions as a sorting adaptor for MyD88.
MyD88, a Toll/interleukin-1 receptor homology (TIR) domain-containing adaptor protein, mediates signals from the Toll-like receptors (TLR) or IL-1/IL-18 receptors to downstream kinases. In MyD88-dependent TLR4 signaling, the function of MyD88 is enhanced by another TIR domain-containing adaptor, Mal/TIRAP, which brings MyD88 to the plasma membrane and promotes its interaction with the cytosolic region of TLR4. Hence, Mal is recognized as the "sorting adaptor" for MyD88. In this study, a direct interaction between MyD88-TIR and another membrane-sorting adaptor, TRAM/TICAM-2, was demonstrated in vitro. Cell-based assays including RNA interference experiments and TRAM deficient mice revealed that the interplay between MyD88 and TRAM in cells is important in mediating IL-18 signal transduction. Live cell imaging further demonstrated the co-localized accumulation of MyD88 and TRAM in the membrane regions in HEK293 cells. These findings suggest that TRAM serves as the sorting adaptor for MyD88 in IL-18 signaling, which then facilitates the signal transduction. The binding sites for TRAM are located in the TIR domain of MyD88 and actually overlap with the binding sites for Mal. MyD88, the multifunctional signaling adaptor that works together with most of the TLR members and with the IL-1/IL-18 receptors, can interact with two distinct sorting adaptors, TRAM and Mal, in a conserved manner in a distinct context.


Comparison of conventional twice daily subcutaneous insulin administration and a multiple injection regimen (using the NovoPen) in insulin-dependent diabetes mellitus.
Thirty-seven stable insulin-dependent diabetic patients receiving twice daily premixed porcine soluble and isophane insulins (Mixtard or Initard, Nordisk Laboratories) were enrolled in a trial to compare the effects of this regimen and an intensified regimen on glycaemic control and flexibility of lifestyle. After a two month run-in period patients were randomized to continue taking premixed insulin (n = 18) or to change to Human Actrapid (three times daily pre-prandial doses using a NovoPen) plus Human Ultratard (n = 19: "pen" group, Novo Laboratories). Diabetic control was assessed at monthly intervals for six months by glycosylated haemoglobin assay (HbA1), 7-point blood glucose profiles and frequency of hypoglycaemic episodes. At randomization, glycaemic control had significantly improved in the group as a whole and patients were well matched with respect to body mass index (BMI) and glycaemic control. No significant changes in BMI, HbA1, mean 7-point glucose profiles or hypoglycaemic episodes were found after six months either within or between groups. Hypoglycaemic episodes were common during the study. There were no technical problems with the NovoPen. Patients using the NovoPen felt it added greater flexibility to their lifestyle. From our study results the main indication to start a multiple injection regimen is to improve the flexibility of the patient's lifestyle and not to improve glycaemic control.


[Late reoperations after repaired Stanford type A aortic dissection].
Objective: To summarize the experience of reoperations on patients who had late complications related to previous aortic surgery for Stanford type A dissection. Methods: From August 2008 to October 2016, 14 patients (10 male and 4 female patients) who underwent previous cardiac surgery for Stanford type A aortic dissection accepted reoperations on the late complications at Department of Thoracic and Cardiovascular Surgery, Nanjing Hospital Affiliated to Nanjing Medical University. The range of age was from 41 to 76 years, the mean age was (57±12) years. In these patients, first time operations were ascending aorta replacement procedure in 3 patients, ascending aorta combined with partial aortic arch replacement in 4 patients, aortic root replacement (Bentall) associated with Marfan syndrome in 3 patients, aortic valve combined with ascending aorta replacement (Wheat) in 1 patient, ascending aorta combined with Sun's procedure in 1 patient, Wheat combined with Sun's procedure in 1 patient, Bentall combined with Sun's procedure in 1 patient. The interval between two operations averaged 0.3 to 10.0 years with a mean of (4.8±3.1) years. The reasons for reoperations included part anastomotic split, aortic valve insufficiency, false aneurysm formation, enlargement of remant aortal and false cavity. The selection of reoperation included anastomotic repair, aortic valve replacement, total arch replacement and Sun's procedure. Results: Of the 14 patients, the cardiopulmonary bypass times were 107 to 409 minutes with a mean of (204±51) minutes, cross clamp times were 60 to 212 minutes with a mean of (108±35) minutes, selective cerebral perfusion times were 16 to 38 minutes with a mean of (21±11) minutes. All patients survived from the operation, one patient died from severe pulmonary infection 50 days after operation. Three patients had postoperative complications, including acute renal failure of 2 patients and pulmonary infection of 1 patient, and these patients were recovered after treatment. Thirteen patients were finally recovered from hospital. The patients were followed up for 16 to 45 months, and no aortic rupture, paraplegia and death were observed in the follow-up. Conclusions: Patients for residual aortic dissection after initial operations on Stanford type A aortic dissection should be attached great importance and always need emergency surgery, but the technique is demanding and risk is great for surgeons and patients, which need enough specification and accurate on aortic operation. More importantly, the Sun's procedure also should be performed on the treatment of residual aortic dissection or distal arch expansion, and obtains the short- and long-term results in the future.


The role of indoleamine 2,3-dioxygenase-aryl hydrocarbon receptor pathway in the TLR4-induced tolerogenic phenotype in human DCs.
A controlled inflammatory response is required for protection against infection, but persistent inflammation causes tissue damage. Dendritic cells (DCs) have a unique capacity to promote both inflammatory and anti-inflammatory processes. One key mechanism involved in DC-mediated immunosuppression is the expression of tryptophan-metabolizing enzyme indoleamine 2,3-dioxygenase (IDO). IDO has been implicated in diverse processes in health and disease but its role in endotoxin tolerance in human DCs is still controversial. Here we investigated the role of IDO in shaping DCs phenotype and function under endotoxin tolerance conditions. Our data show that TLR4 ligation in LPS-primed DCs, induced higher levels of both IDO isoforms together with the transcription factor aryl-hydrocarbon receptor (AhR), compared to unprimed controls. Additionally, LPS conditioning induced an anti-inflammatory phenotype in DCs - with an increase in IL-10 and higher expression of programmed death ligand (PD-L)1 and PD-L2 - which were partially dependent on IDO. Furthermore, we demonstrated that the AhR-IDO pathway was responsible for the preferential activation of non-canonical NF-κB pathway in LPS-conditioned DCs. These data provide new insight into the mechanisms of the TLR4-induced tolerogenic phenotype in human DCs, which can help the better understanding of processes involved in induction and resolution of chronic inflammation and tolerance.


Bone marrow transplantation for inherited diseases.
Allogeneic bone marrow transplantation has been used successfully for the treatment of a variety of inherited diseases. The goal of transplantation in this setting is to provide a sufficient degree of sustained marrow engraftment to allow longterm amelioration of the inherited disease phenotype. Many factors influence the likelihood of achieving this goal, including donor availability, conditioning regimen, marrow processing, and the nature and extent of progression of the disease. For many inherited diseases early diagnosis is imperative because the outcome of transplantation is more favorable when performed prior to the development of significant organ damage from the disease, its complications, or treatment. Although the results of bone marrow transplantation are good for some inherited diseases and are improving for others, significant problems remain such as donor availability, conditioning regimen toxicity, graft failure, and graft-versus-host disease. This review describes some of the unique features of bone marrow transplantation for inherited diseases and discusses recent advances in this area.


Studies of the efficacy of a novel intranasal vaccine against feline bordetellosis.
A novel intranasal vaccine against disease caused by Bordetella bronchiseptica in cats was tested in a series of three experiments. In the first experiment a vaccinated group and an unvaccinated control group of kittens were challenged by the aerosol route with virulent B bronchiseptica three weeks after they had been vaccinated. The control kittens developed upper respiratory tract signs typical of feline B bronchiseptica infection, including rhinitis, a serous ocular and nasal discharge, fever, sneezing and coughing. The mean (sd) clinical score for the cats in the unvaccinated control group was 19.5 (5.4) compared with 1.53 (1.9) for the vaccinated group. In the second experiment vaccinated kittens were challenged with virulent B bronchiseptica 72 hours after they were vaccinated. Their mean clinical score was 2.76 (2.62) compared with 13.4 (3.33) for the control group. In the final experiment, vaccinated and unvaccinated control cats were challenged after six or 12 months. After six months the mean clinical scores were 13.9 (4.7) for the control group, compared with 1.33 (1.56) for the vaccinated group, and after 12 months the scores were 9.92 (5.79) for the control group compared with 0.92 (0.89) for the vaccinated group.


[Correlation analysis on the malocclusion and articulation of skeletal angle III malocclusion in mixed dentition].
To investigate the effects of skeletal Class III malocclusion in mixed dentition on speech articulation and to look for which factors lead to the speech errors. Thirty-eight children with skeletal Angle Ill malocclusion in mixed dentition were selected as a sample group and 40 children with normal occlusion in mixed dentition as a control group. Two phoneticians evaluated their articulations and wrote down error phonemes respectively. The correlation analysis was undertaken between the number of errors and the measurements of patients' cephalometry. The number of errors were correlated significantly with overbite, UI-LI, OBJ (OB+OJ) and TD-PW. There is articulatory malfunction in the majority of skeletal Angle III malocclusion patients in mixed dentition. Articulatory malfunction is related to the position of incisors and the tongue.


Active nucleolus organizers are precisely positioned in adult central nervous system cells but not in neuroectodermal tumor cells.
Active nucleolus organizing regions (NOR) were stained with silver in isolated nuclei and chromosomes, monolayer cultures, and tissues for electron microscopy. In nuclei isolated in 0.8-1.3 M urea, only the NOR stained with silver, and round or fiber-like regions with a diameter of 200-250 nm with a substructure were delineated at appropriate nucleolar or acrocentric chromosomal sites. In whole cells, additional (non NOR) silver binding regions were noted. Electrophoresis studies on nuclear isolates indicated at least two different nuclear protein subsets were responsible for the observed silver binding; the NOR protein(s) was most tightly bound with respect to urea. Interphase cells displayed a more extensive NOR network than mitotic cells, suggesting an increase in rDNA transcription during interphase. Mature sperm cells showed no active NOR regions. Interphase neuroectodermal tumor cells generally contained large multiple NOR indicating extensive activity; the position of NOR in the nucleus was variable from cell to cell, even in cloned lines. In contrast to tumor lines, central nervous system (CNS) cells from various species all showed highly reproducible or non-random NOR locations within the nucleus of each cell type. The NOR of large neurons were always central and single, whereas small granule cell neurons displayed a few small NOR that were positioned more peripherally. These findings suggest that in highly differentiated cells, the NOR region is precisely positioned in the nucleus, regardless of non-identical chromosome locations of the NOR in different species. The variability of NOR in tumor cells indicates profound changes in nuclear structure that may be part of the neoplastic transformation.


Phenotype-associated lectin-binding profiles of normal and transformed blood cells: a comparative analysis of mannose- and galactose-binding lectins from plants and human serum/placenta.
Surface glycoconjugates of normal and transformed blood cells are commonly characterized by plant lectins. To infer physiological significance of protein-carbohydrate interactions, mammalian lectins are obviously preferable as research tools. So far, human serum lectins have not been used to assess their binding to immunophenotyped human normal or transformed blood cells. Thus, our study combines two groups of lectins with different specificity from plant and human sources. Besides concanavalin A (ConA) we have isolated the mannose-binding protein and serum amyloid P component from human serum. Especially the mannose-binding protein is believed to play a role in host defence against bacteria and yeast cells with unknown impact on normal and tumor cells. These three lectins establish the first group. In addition to the immunomodulatory mistletoe lectin, whose binding can elicit enhanced cytokine secretion from mononuclear blood cells, we included the beta-galactoside-binding lectin (14 kDa) from human placenta in the second group. The initial series of measurements was undertaken using two-color flow cytometry to determine the phenotype-associated binding (based on cluster designation; CD) of the lectins to blood and bone marrow cells from normal donors and the cell line CEM (T-lymphoblastoid), KG1-A (primitive myeloid leukemia) and Croco II (B-lymphoblastoid). Heterogeneity was apparent for each lectin in the CD-defined cell populations. Significant differences in binding were noted between Viscum album agglutinin (VAA) and other lectins for CD4+ cells from blood and between mannose-binding protein (MBP) and VAA versus 14 kDa, ConA and serum amyloid P component (SAP) for CD19+ cells from bone marrow.(ABSTRACT TRUNCATED AT 250 WORDS)


Catalytic, Enantioselective, Intramolecular Sulfenofunctionalization of Alkenes with Phenols.
The catalytic, enantioselective, cyclization of phenols with electrophilic sulfenophthalimides onto isolated or conjugated alkenes affords 2,3-disubstituted benzopyrans and benzoxepins. The reaction is catalyzed by a BINAM-based phosphoramide Lewis base catalyst which assists in the highly enantioselective formation of a thiiranium ion intermediate. The influence of nucleophile electron density, alkene substitution pattern, tether length and Lewis base functional groups on the rate, enantio- and site-selectivity for the cyclization is investigated. The reaction is not affected by the presence of substituents on the phenol ring. In contrast, substitutions around the alkene strongly affect the reaction outcome. Sequential lengthening of the tether results in decreased reactivity, which necessitated increased temperatures for reaction to occur. Sterically bulky aryl groups on the sulfenyl moiety prevented erosion of enantiomeric composition at these elevated temperatures. Alcohols and carboxylic acids preferentially captured thiiranium ions in competition with phenolic hydroxyl groups. An improved method for the selective C(2) allylation of phenols is also described.


Hormetic approaches to the treatment of Parkinson's disease: Perspectives and possibilities.
Age-related changes in the brain reflect a dynamic interaction of genetic, epigenetic, phenotypic, and environmental factors that can be temporally restricted or more longitudinally present throughout the lifespan. Fundamental to these mechanisms is the capacity for physiological adaptation through modulation of diverse molecular and biochemical signaling occurring from the intracellular to the network-systemic level throughout the brain. A number of agents that affect the onset and progression of Parkinson's disease (PD)-like effects in experimental models exhibit temporal features, and mechanisms of hormetic dose responses. These findings have particular significance since the hormetic dose response describes the amplitude and range of potential therapeutic effects, thereby affecting the design and conduct of studies of interventions against PD (and other neurodegenerative diseases), and may also be important to a broader consideration of hormetic processes in resilient adaptive responses that might afford protection against the onset and/or progression of PD and related disorders.


Pulsed Dilution Method for the Recovery of Aggregated Mouse TNF-α.
The expression of mouse tumor necrosis factor alpha (TNF-α) in Escherichia coli is a favorable way to get high yield of protein; however, the formation of cytoplasmic inclusion bodies, which is the consequence of insoluble accumulated proteins, is a major obstacle in this system. To overcome this obstacle, we used a pulsed dilution method to convert the product to its native conformation. Reducing agent and guanidine hydrochloride were used to solubilize inclusion bodies formed after TNF-(α) expression. Then, the refolding procedure was performed by pulsed dilution of the denatured protein into a refolding buffer. The properly-folded protein was purified by metal affinity chromatography. SDS-PAGE showed a 19.9 kDa band related to the mature TNF-(α) protein. The protein was recognized by anti-mouse TNF-(α) on western blots. The final concentration of the purified recombinant TNF-(α) was 62.5 µg/mL. Our study demonstrates the efficiency of this method to produce a high yield of folded mature TNF- (α).


Does the Level of Difficulty in Balancing Tasks Affect Haptic Sensitivity Via Light Touch?
The purpose of this study was to determine whether the contribution of bimanual light touch varies according to the difficulty level of postural tasks (e.g., vision occlusion, height of support surface). Fourteen healthy young adults each were asked to stand in a tandem position, on a 20-cm height balance beam. Postural tasks included light touch and no touch conditions in two vision conditions, nonvision and full vision. The root mean square of amplitude of oscillation (mediolateral), mean velocity, ellipse area, and path length of the center of pressure revealed that touch conditions reduced sway to a greater extent in the elevated support surface, nonvision condition. Highly unstable balance tasks increase the optimization of light touch and affect the attenuation of postural sway.


Correlation between the Flow Temperature and Mineral Factor for Coal Ashes Based on FactSage Calculation.
To find a simple method to predict the ash flow temperature (FT) based on FactSage calculation, 69 coal ash samples were selected to explore the correlation between the FT and its mineral factor (MF) at a given temperature. An approximate linear relationship was found between ash FT and its MF: FT = 0.64MF + 1332, with a related coefficient value of 0.94 and a standard deviation of 25.77 °C. Ten ash samples were used to investigate its reliability. The calculated FTs were consistent with experimental FTs in the error range of measurement, and the correlation might be more reliable for low iron and low calcium coal.


Naturally occurring xanthone and benzophenone derivatives exert significant anti-proliferative and proapoptotic effects in vitro on synovial fibroblasts and macrophages from rheumatoid arthritis patients.
There is a need for novel, safer and cheaper drugs for the therapy of rheumatoid arthritis (RA), better targeted against the cellular processes involved in the disease pathogenesis. Using advanced analysis of microscopic images and flow cytometry, we demonstrate that naturally occurring xanthone and benzophenone derivatives exert strong, dose- and O2 concentration-dependent anti-proliferative and pro-apoptotic effects on RA patients' fibroblast-like synoviocytes (FLS) and macrophages. Suspensions containing fibroblasts, macrophages and other infiltrating cells were obtained from inflamed synovial tissue collected from female RA patients. Cells were grown in the presence of xanthone (mangiferin, isomangiferin, neomangiferin, norathyriol) or benzophenone (iriflophenone 3-C-glucoside, maclurin) derivatives for 48h or 7days, at 5% or 21% O2. Proportions of macrophages, FLS and infiltrating T cells undergoing apoptosis (annexin- or annexin and 7-AAD-positive) were determined by flow cytometry. The extent of late apoptosis (DNA degradation) was assessed by fluorescent microscopy and image analysis in cultures where DNA was stained with Hoechst 33342. Majority of tested compounds exert anti-proliferative and pro-apoptotic, O2-dependent effects on T cells, FLS and macrophages. The results indicate that xanthone- and benzophenone-rich plant products provide a basis for the development of dietary strategy for rheumatoid arthritis management.


Global pharmacogenetics: genetic substructure of Eurasian populations and its effect on variants of drug-metabolizing enzymes.
To study the frequency distribution of cytochrome P450 (CYP) functional genetic variants in five Eurasian populations from the territory of Siberia in Russia. Unrelated healthy Tuvinians, Buryats, Altaians, Yakuts and Russians (n = 87-88) were genotyped for CYP2C9*2, CYP2C9*3, CYP2C19*2, CYP2C19*3, CYP3A5*3 and CYP3A5*6. Standard pairwise genetic distances, locus-specific and global Fst statistics were calculated. CYP allele and genotype frequencies demonstrated significant variability. Overall, the degree of between-population variance displayed by CYP SNPs was lower than that recorded from neutral short tandem repeats and Alu-insertion polymorphism, indicating evolutionary conservation of CYP polymorphisms. CYP-based genetic distances were well correlated with the geographic distances across populations (r = 0.822, p = 0.008). Although the tested variants were present in the neighboring, yet secluded, populations at the expected range of frequencies, the observed frequencies were significantly variable across Eurasian populations, indicating potential relevance to clinical decision making.


The histopathologic spectrum of carcinomas involving the gastroesophageal junction in the Chinese.
Carcinomas involving the gastroesophageal junction are common in China. The histopathologic characteristics of these cancers have not been systematically investigated. Reported are 41 such resected cancers from Chinese patients (30 men, 11 women). Their mean age was 62 years. The mean tumor size was 4.4 cm (range, 2 to 9 cm), and 58% were poorly differentiated. An unusual spectrum of tumor differentiation was observed, including adenocarcinomas (83%), adenosquamous (32%), colloid (2%), signet-ring (10%), squamous (5%), oncocytic (7%), pancreatic acinar (12%), and neuroendocrine (5%) carcinomas. Cancers with multiple types of differentiation in the same tumor were identified in 37 cases (90%). The adjacent gastric cardiac mucosa showed hyperplasia, oncocytic, and pancreatic acinar metaplasia, and mild chronic inflammation. Dysplasia was uncommon (n = 6). Barrett esophagus was not identified. Carcinomas involving the gastroesophageal junction in the Chinese are morphologically distinct, heterogeneous, and may be of esophageal origin.


The periaqueductal gray and Bayesian integration in placebo analgesia.
In placebo hypoalgesia research, the strength of treatment expectations and experiences are key components. However, the reliability or precision of expectations had been mostly ignored although being a likely source for interindividual differences. In the present study, we adopted a Bayesian framework, naturally combining expectation magnitudes and precisions. This postulates that expectations (prior) are integrated with incoming nociceptive information (likelihood) and both are weighted by their relative precision to form the pain percept and placebo effect. Sixty-two healthy subjects received heat pain during fMRI. Placebo effects were more pronounced in subjects with more precise treatment expectations and correlated positively with the relative precision of the prior expectation. Neural correlates of this precision were observed in the periaqueductal gray and the rostral ventromedial medulla, indicating that already at the level of the brainstem the precision of an expectation can influence pain perception presenting strong evidence for Bayesian integration in placebo hypoalgesia.


Single-cell ATAC-seq signal extraction and enhancement with SCATE.
Single-cell sequencing assay for transposase-accessible chromatin (scATAC-seq) is the state-of-the-art technology for analyzing genome-wide regulatory landscapes in single cells. Single-cell ATAC-seq data are sparse and noisy, and analyzing such data is challenging. Existing computational methods cannot accurately reconstruct activities of individual cis-regulatory elements (CREs) in individual cells or rare cell subpopulations. We present a new statistical framework, SCATE, that adaptively integrates information from co-activated CREs, similar cells, and publicly available regulome data to substantially increase the accuracy for estimating activities of individual CREs. We demonstrate that SCATE can be used to better reconstruct the regulatory landscape of a heterogeneous sample.


Prophylactic effects of magnesium and vitamin E in rat spinal cord radiation damage: evaluation based on lipid peroxidation levels.
This study investigated the neuroprotective effects of magnesium sulfate prophylaxis and vitamin E prophylaxis in a rat model of spinal cord radiation injury. Groups were subjected to different treatment conditions for 5 days prior to irradiation, and outcomes were evaluated on the basis of lipid peroxidation levels in cord tissue. Four groups of rats were investigated: no radiation/treatment (n = 4), intraperitoneal (i.p.) saline 1 ml/day (n = 6), i.p. vitamin E 100 mg/kg/day (n = 6), and i.p. magnesium sulfate 600 mg/kg/day (n = 6). The thoracic cord of each non-control rat was exposed to 20 Gy radiation in a LINAC system using 6 MV x-rays, and malondialdehyde (MDA) levels (reflecting lipid peroxidation level) were determined 24 hours post-irradiation. The MDA levels in thoracic cord segments from the control rats were used to determine baseline lipid peroxidation. The mean levels in the control, saline-only, vitamin E, and magnesium sulfate groups were 12.12 +/- 0.63, 27.0 +/- 2.81, 17.71 +/- 0.44, and 14.40 +/- 0.47 nmol/mg tissue, respectively. The MDA levels in the saline-only group were significantly higher than baseline, and the levels in the vitamin E group were significantly lower than those in the saline group (P < 0.05 for both). The levels in the magnesium sulfate group were dramatically lower than those in the saline group (P < 0.001). The results indicate that i.p. magnesium sulfate has a marked neuroprotective effect against radiation-induced oxidative stress in the rat spinal cord.


Some observations on protease production in continuous suspension cultures of Bacillus firmus.
Invariance of culture conditions in steady state continuous cultures make these a very valuable tool to study the influence of various culture parameters on cell growth and synthesis of primary and secondary metabolites. The result of a parametric study on production of protease in continuous suspension cultures of Bacillus firmus NRS 783 are reported in this article. This strain is a superior producer of an alkaline protease with major application in the detergent industry. The parameters investigated include dilution rate and concentrations of yeast extract, ammonium, and inorganic phosphate in the bioreactor feed, glucose being the principal carbon source in all experiments. The regulatory effects of the key culture parameters on cell growth, synthesis and secretion of protease, and production of acetic acid are investigated. The relations among the specific cell growth rate, specific utilization rates of the principal carbon, nitrogen, and phosphorous sources, and specific production rates of two nonbiomass products, viz., acetic acid and protease, are examined, and the effects of the manipulated culture parameters on these relations, specific protease activity, and yields of cell mass, protease, and acetic acid on the basis of the principal carbon, nitrogen, and phosphorous sources are studied. An increase in dilution rate led to increases in specific utilization rates of the principal carbon, nitrogen, and phosphorous sources and specific production rates of acetic acid and protease and decreases in bulk activities/concentrations of the three products (acetic acid, cell mass, and protease). As a result, the productivities of the three species were maximized at an intermediate dilution rate. Increased supply of yeast extract (a rich source of amino acids, proteins, and vitamins, besides being an additional source of carbon, nitrogen, and phosphorus) promoted cell mass formation but reduced protease production per unit cell mass. Increased supply of nitrogen and phosphorous sources stimulated protease synthesis up to certain threshold levels and repressed the enzyme synthesis beyond the threshold levels. With increased supply of the nitrogen source, the phosphorous source was more efficiently utilized for cell growth and protease synthesis. Stable maintenance of continuous cultures of B. firmus over prolonged period is demonstrated in this study.


Localization of Inv in a distinctive intraciliary compartment requires the C-terminal ninein-homolog-containing region.
The primary cilium is an antenna-like structure extending from the surface of most vertebrate cells. Loss or mutation of ciliary proteins can lead to polycystic kidney disease and other developmental abnormalities. inv mutant mice develop multiple renal cysts and are a model for human nephronophthisis type 2. The mouse Inv gene encodes a 1062-amino-acid protein that is localized in primary cilia. In this study, we show that the Inv protein (also known as inversin) is localized at a distinctive proximal segment of the primary cilium, using GFP-tagged Inv constructs and anti-Inv antibody. We named this segment the Inv compartment of the cilium. Further investigation of the Inv protein showed that 60 amino acids at its C-terminal, which contains ninein homologous sequences, are crucial for its localization to the Inv compartment. Fluorescence recovery after photobleaching analysis revealed that the Inv protein was dynamic within this compartment. These results suggest that localization of the Inv protein to the Inv compartment is actively regulated. The present study revealed that the primary cilium has a distinct molecular compartment in the body of the primary cilium with a specific confining and trafficking machinery that has not been detected previously by morphological examination.


BDNF serum levels are not related to cognitive functioning in older depressed patients and controls.
Depression and cognitive decline are highly prevalent in older persons and both are associated with low serum brain derived neurotrophic factor (BDNF). Mutual pathways of depression and cognitive decline in older persons may explain the overlap in symptoms and low serum BDNF. We hypothesized that serum BDNF levels are lower in depressed elderly with poor cognitive performance (global or specifically in working memory, speed of information processing, and episodic memory) compared to depressed elderly without cognitive impairment or non-depressed controls. BDNF Serum levels and cognitive functioning were examined in 378 depressed persons and 132 non-depressed controls from a large prospective study on late-life depression. The association between BDNF levels and each cognitive domain among the depressed patients was tested by four separate linear regression models adjusted for relevant covariates. An analysis of covariance (ANCOVA) was performed to compare BDNF serum levels in three groups (depression with cognitive impairment, depression without cognitive impairment, and non-depressed controls), when adjusted for potential confounders. No significant linear association was found between BDNF and any of the four cognitive domains tested. There are no differences in BDNF levels between controls and depressed patients with or without cognitive impairment global or in specific domains after controlling for confounders. BDNF serum levels in this cohort of older depressed patients and controls are not related to cognitive functioning. As BDNF is essential for the survival and functioning of neurons, its levels may remain normal in stages of disease where remission is achievable.


Extracts of Larix Leptolepis effectively augments the generation of tumor antigen-specific cytotoxic T lymphocytes via activation of dendritic cells in TLR-2 and TLR-4-dependent manner.
Type-1 immunity plays a crucial role in host defense against various tumors and infectious diseases. Here, we first demonstrated that extract of Larix Leptolepis (ELL), one of the most popular timbers at Hokkaido area in Japan, strongly activated Type-1 immunity. ELL induced production of Type-1 cytokines such as IL-12 and TNF-α from bone marrow-derived dendritic cells (BMDCs) in TLR2- and TLR4-dependent manner and remarkably up-regulated the expression of MHC and co-stimulatory molecules. In addition, antigen-specific CTLs were significantly augmented by the combined administration of ELL, antigen and BMDCs. Finally, we revealed that combination therapy using ELL, antigen and BMDCs significantly inhibited the growth of established tumor in mouse model. Thus, these findings suggested that ELL would be a novel adjuvant for inducing an activation of Type-1-dependent immunity including activation of BMDCs and induction of tumor-specific CTLs, which is applicable to the therapy of cancer and infectious diseases.


QT Dispersion in Healthy Adult Nigerians.
Increase in QT dispersion (QTd) is associated with an increased risk of cardiovascular morbidity and mortality. This study sought to (i) determine the mean QTd and (ii) characterise QTd in a healthy Nigerian population. One hundred healthy Nigerian adults were studied. Healthy status of the subjects was determined by history and physical examination. A resting 12- lead ECG was obtained from all subjects for determination of QTc, QTd and ECG left ventricular hypertrophy (LVH) using Sokolow Lyon (SL) and Araoye's codes. Echocardiography was used to determine LV systolic function, LVM and LVMI for 60 subjects. The QTd ranged from 15-70ms with a mean value of 38.5 ± 11.2ms. QTd was independent of age (p = 0.86), sex (p = 0.97), heart rate (p = 0.22), blood pressure (p > 0.05), BMI (p = 0.81), QTc (p = 0.41), LVH (ECG and echo) and LV systolic function (p > 0.05). QT dispersion is independent of demographic parameters, LV systolic function and LV hypertrophy in healthy adult Nigerians.


Potential of electrospray mass spectrometry for quantifying glycohemoglobin.
An electrospray ionization-mass spectrometric procedure has been developed for determining glycohemoglobin. Whole-blood samples from 78 diabetic and 50 nondiabetic subjects (glycation range 3-15%, as determined by electrospray mass spectrometry) were diluted 500-fold in an acidic denaturing solvent and introduced directly into a mass spectrometer. The resulting mass spectra were then processed to estimate the percentage of glycohemoglobin present in the sample. Total analysis time, including plotting the spectra and computing the percentage of glycation, was approximately 3 min. The imprecision (CV) of the method was < 5.1% for inter- and intrabatch analyses for total glycohemoglobin in the range 3.6-14%. Comparison of the mass spectrometric results with those from established affinity chromatographic procedures showed good overall agreement. The relative glycation of the alpha- and beta-chains was determined directly and was shown to be constant (0.64:1) over the glycation range measured. Only single glucose attachment to both the alpha- and beta-chains was observed.


Young adult reference ranges for thyroid function tests on the Centaur immunoassay analyser.
This study aims to establish reference ranges for thyroid tests in young Saudi adults using the Centaur immunoassay method. Physical examination is performed and thyroid function tests include thyroid stimulating hormone (TSH), free thyroxine (FT4) and free triiodothyronine (FT3). These are performed on 291 young Saudi adults (182 [63%] females and 109 [37%] males; average age: 27 years [range 18-50]). Clinical thyroid abnormality, related symptoms and/or abnormal thyroid function tests exclude a person from the study and thus a total of 276 subjects (171 [62%] females and 105 [38%] males) are used to establish the new reference ranges. Combined female and male ranges for TSH, FT4, and FT3 were found to be 0.48-6.30 miu/L (9.00-18.62 pmol/L and 3.39-6.85 pmol/L, respectively). Mean TSH and FT4 levels were significantly different (P<0.0001) from those quoted by the manufacturer. Ranges for TSH were 0.48-6.30 miu/L (female) and 0.52-4.89 miu/L (male) (P=0.08). Female ranges for FT4 and FT3 were 9.00-17.15 pmol/L and 3.39-5.82 pmol/L, respectively. Male ranges were 9.92-18.62 pmol/L (P=0.0001) and 4.36-6.85 pmol/L (P<0.0001). The range of TSH levels in the young local Saudi population proved to be higher than that quoted by the manufacturer. FT4 range was lower and narrower than that quoted by the manufacturer. Significant differences between female and male populations suggest that partitioning of the reference ranges by gender is necessary.


Plant proteome analysis: a 2004-2006 update.
Since the appearance of the review entitled "Plant Proteome Analysis" in Proteomics in February 2004 (Cánovas, F. M., Dumas-Gaudot, E., Recorbert, G., Jorrín, J. et al., Proteomics 2004, 4, 285-298), about 200 original articles focusing on plant proteomics have been published. Although this represents less than 1% of the global proteomics output during this period, it nevertheless reflects an increase in activity over the period 1999-2004. These papers concern the proteome of at least 35 plant species but have concentrated mainly on thale cress (Arabidopsis thaliana) and rice (Oryza sativa). The scientific objectives have ranged from a proteomic analysis of organs, tissues, cell suspensions, or subcellular fractions to the study of plant development and response to various stresses. A number of contributions have covered PTMs and protein interactions. The dominant analytical platform has been 2-DE coupled to MS, but "second generation" techniques such as DIGE, multidimensional protein identification technology, isotope-coded affinity tags, and stable isotope labeling by amino acids in cell culture have begun to make an impact. This review aims to provide an update of the contribution of proteomics to plant biology during the period 2004-2006, and is divided into six sections: introduction, subcellular proteomes, plant development, responses to biotic and abiotic stresses, PTMs, and protein interactions. The conclusions summarize a view of the major pitfalls and challenges of plant proteomics.


Leukocyte filtration in lung transplantation.
Controlled reperfusion of the transplanted lung has been used in nine consecutive patients to decrease manifestations of lung reperfusion injury. An extracorporeal circuit containing a roller pump, heat exchanger and leukodepleting filter is primed with substrate-enhanced reperfusion solution mixed with approximately 2000 ml of the patient's blood. This solution is slowly recirculated to remove leukocytes prior to reperfusion. When the pulmonary anastomoses are completed, the pulmonary artery is cannulated through the untied anastomosis using a catheter containing a pressure lumen for measurement of infusion pressure. An atrial clamp is left in place on the patient's native atrial cuff to decrease the risk of systemic air embolism during the brief period of reperfusion from the extracorporeal reservoir. During reperfusion, the water bath to the heat exchanger is kept at 35 degrees C and the flow rate for reperfusion solution is between 150 and 200 m/min, keeping the pulmonary artery pressure <14 mmHg. Eight of nine patients were ventilated on 40% inspired oxygen within a few hours of operation and 7/9 were extubated on or before postoperative day 1. Six of nine patients are long-term survivors.


Palladium-catalyzed stereoselective construction of chiral allenes bearing nonadjacent axial and two central chirality.
It is challenging to enantioselectively construct molecules bearing multiple nonadjacent stereocenters, in contrast to those bearing a single stereocenter or adjacent stereocenters. Herein, we report an enantio- and diastereoselective synthesis of substituted chiral allenes with nonadjacent axial and two central chiral centers through a combination of retro-oxa-Michael addition and palladium-catalyzed asymmetric allenylic alkylation. This methodology exhibits good functional-group compatibility, and the corresponding allenylic alkylated compounds, including flavonoid frameworks, are obtained with good yields and diastereoselectivities and excellent enantioselectivities (all >95% ee). Furthermore, the scalability of the current synthetic protocol was proven by performing a gram-scale reaction.


Prognostic Value and Daily Trend of Interleukin-6, Neutrophil CD64 Expression, C-Reactive Protein and Lipopolysaccharide-Binding Protein in Critically Ill Patients: Reliable Predictors of Outcome or Not?
Severe sepsis and/or trauma complicated by multiple organ dysfunction syndrome are the leading causes of death in critically ill patients. The aim of this prospective single-centre study was to assess the prognostic value and daily trend of interleukin-6 (IL-6), neutrophil CD64 expression, C-reactive protein (CRP) and lipopolysaccharide-binding protein (LBP) regarding outcome in critically ill patients with severe trauma and/or severe sepsis. Outcome measure was hospital mortality. One hundred and two critically ill patients admitted to the intensive care unit of a tertiary university hospital were enrolled in this prospective study. Blood samples were collected on admission (day 1), days 2 and 3. CD64 index was 1.6-fold higher on day 1 and 1.78-fold higher on day 2 in non-survivors (p<0.05). The area under the curve (AUC) for the CD64 index on day 1 for outcome was 0.727. At a cut-off level of 2.80 sensitivity was 75% and specificity was 65%. Patients with CD64 index level on day 1 higher than 2.80 had 2.4-fold higher probability of dying. Odds ratio is 2.40; 95% CI 0.60-9.67. CD64 index on day 1 is a fairly good predictor of outcome. AUCs for IL-6, CRP and LBP were < 0.55, suggesting these biomarkers failed to predict outcome.


The Dorsal Frontoparietal Network: A Core System for Emulated Action.
The dorsal frontoparietal network (dFPN) of the human brain assumes a puzzling variety of functions, including motor planning and imagery, mental rotation, spatial attention, and working memory. How can a single network engage in such a diversity of roles? We propose that cognitive computations relying on the dFPN can be pinned down to a core function underlying offline motor planning: action emulation. Emulation creates a dynamic representation of abstract movement kinematics, sustains the internal manipulation of this representation, and ensures its maintenance over short time periods. Based on these fundamental characteristics, the dFPN has evolved from a pure motor control network into a domain-general system supporting various cognitive and motor functions.


[Cardiac operations in nine patients aged 80 years or older].
Nine patients aged 80 years or older underwent cardiac operations with cardiopulmonary bypass. Seven patients underwent coronary artery bypass surgery, one modified Bentall's operation and another patch closure of ventricular septal perforation. One patient died soon after operation (11 POD) and operative mortality was 11%. One died 7 months after operation because of rupture of dissecting thoracic aortic aneurysm. Seven patients are still alive within physical status of New York Heart Association Classes I or II. We conclude that cardiac operations should not be excluded in octogenarians because of their chronological age alone.


Family planning in a rural setting in Uganda, the USHAPE initiative.
The total fertility rate in Uganda is 5.9 children per woman, and women admit to having nearly two more children than they actually want. The maternal mortality rate remains stubbornly high. Family planning saves lives. It prevents maternal deaths by delaying motherhood, helping women limit their family size and avoid unwanted pregnancies. It also reduces infant mortality. USHAPE (Ugandan Sexual Health and Pastoral Education) is an initiative run in conjunction with the Royal College of General Practitioners in south-west Uganda. USHAPE aims to disseminate positive messages about modern contraception in an attempt to dispel fears and misconceptions and address the high rate of unmet need. The aim was to determine the rate of unmet need for family planning among women of reproductive age in the population local of Kisiizi hospital and to use the successful USHAPE model to train health workers to address this need. 100 patients were screened in the outpatient department to determine the level of unmet need by asking 2 questions. Level 1 training aims enhance every staff member's knowledge, so that the responsibility for family planning is adopted by the whole institution. Level 2 trains clinicians to become full family planning providers, with the necessary communication, educational and practical skills. The screening for unmet need for contraception revealed that 51% have an unmet need, which is higher than the national average of 38%. Sixty-eight members of staff at Kisiizi trained to a basic level and a further 32 staff have been trained to Level 2 higher level. The USHAPE approach has begun to tackle some of the barriers to accessing family planning, but there are further areas which need development. Our cascade model of training, involves training Ugandan USHAPE trainers with the aim of future scale up and long-term development.


Modifications by ischemia of carbohydrate and lipid metabolism of the dog heart in situ from circulating and endogenous substrates.
The uptake of circulating substrates, lactate, glucose and free fatty acids (FFA) has been investigated concurrently with the tissular contents of these principles and the glycogen and triglyceride stores in the dog heart in situ submitted to incomplete obstruction of left coronary bed. Transmural samples necessary for the repeated determination of tissular substrates were taken from left ventricular wall by means of a total cardiopulmonary by-pass system, then divided to allow the analysis separately in subendocardial and subepicardial layer. A 40 to 70% reduction in coronary blood flow gave rise to decrease or suppression of uptake of all the substrates or even to conversion of uptake into output. The modifications of uptake are chiefly related to the deficiency of breakdown by oxidation, though lessened in the case of FFA by incorporation into triglycerides and enhanced in the case of glucose by glycogenolysis. Glycogenolysis and consequent anaerobic glycolysis appear to be the main process available against the energy cellular defect linked with oxygen lack which affects notably more subendocardial than subepicardial layer.


Efficient large-scale screening of viral pathogens by fragment length identification of pooled nucleic acid samples (FLIPNAS).
The necessity for the large-scale screening of viral pathogens has been amply demonstrated during the COVID-19 pandemic. During this time, SARS-CoV-2 nucleic acid pooled testing, such as Dorfman-based group testing, was widely adopted in response to the sudden increased demand for detection. However, the current approach still necessitates the individual retesting of positive pools. Here, we established an efficient method termed the fragment-length identification of pooled nucleic acid samples (FLIPNAS), where all subsamples (n = 8) can be uniquely labelled and tested in a single-time detection among pools of samples. We used a novel and simple design of unique primers (UPs) to generate amplicons of unique lengths after reverse transcription and polymerase chain reaction to reach this aim. As a result, the unique lengths of the amplicons can be recognized and traced back to the corresponding UPs and specific samples. Our results demonstrated that FLIPNAS could recognize one to eight positive subsamples in a single test without retesting positive pools. The system also showed sufficient sensitivity for the mass monitoring of SARS-CoV-2 and no cross-reactivity against three common respiratory diseases. Moreover, the FLIPNAS results of 40 samples with a positive ratio of 7.8% were in 100% agreement with their individual detection results using the gold standard. Collectively, this study shows that the efficiency of nucleic acid pooling detection can be further improved by FLIPNAS, which can speed up testing and mitigate the urgent demand for resources.


[The predictive value of DE-CMR in patients with severe chronic aortic regurgitation and extremely dilated left ventricular chamber].
To determine whether preoperative contrast delay-enhanced cardiovascular magnetic resonance imaging (DE-CMR) could help predict long-term survival of patients with severe chronic aortic regurgitation and extremely dilated left ventricular chamber after aortic valve replacement. Totally 37 patients enrolled between February 2008 and November 2010 with severe chronic aortic regurgitation and extremely dilated left ventricular chamber, who met the echo criteria, that was left ventricular end diastolic dimension > 70 mm or left ventricular end systolic dimension > 55 mm, and were scheduled to the surgery. The 2-dimensional echocardiographic examinations and CMR with late gadolinium-enhancement (LGE) were performed routinely preoperatively. According to the results of CMR, the patients were divided into 2 groups: the LGE positive(+) group and LGE negative(-) group. The association of LGE with event free survival, postoperative cardiac function and postoperative hospital stay time was investigated. Fifteen patients had significant LGE signals in CMR films, while the other twenty-two were silent. All of them received the operative procedures, including aortic valve replacement in 28 cases, Bentall procedure in 3 cases, aortic valve replacement and ascending aorta replacement in 6 cases, and concomitant mitral valve repair in 11 cases. Over a follow-up of 33.6 months, 1-year, 2-year and 3-year event free survival rates in LGE(-) group were 94.7%, 88.4%, and 72.6%, respectively, compared to 80.0%, 48.9%, and 32.6%, respectively in LGE(+) group (χ(2) = 7.244, P = 0.007). The postoperative hospital stay time of LGE(-) group was (9 ± 2) days, which of LGE(+) group was (10 ± 3) days (t = 1.175, P = 0.248). LGE positive signal in CMR films is a potential predictor of persistent cardiac failure after aortic valve replacement for patients with severe chronic aortic regurgitation and extremely dilated left ventricular chamber. It has intimate relationship with malignant arrhythmia and sudden death, which makes it a valuable technique in preoperative evaluation and risk stratification.


Prodromal spinocerebellar ataxia type 2: Prospects for early interventions and ethical challenges.
The characterization of prodromal stages in neurodegenerative disorders is becoming increasingly important because of the need for early neuroprotective therapies. Research during the past 3 decades in spinocerebellar ataxia type 2 has revealed a large body of evidence suggesting that many disease features precede the manifest cerebellar syndrome, which delineates the prodromal stage of this disorder. This stage is defined by clinical, imaging, and functional criteria, which are supported by early molecular events demonstrated in animal models. Knowledge regarding prodromal spinocerebellar ataxia type 2 provides insight into the mechanisms underlying neurodegeneration from the early stages, which enables the design of promising disease-modifying clinical trials through the identification of the optimum moment to begin the therapies, the appropriate selection of individuals, and the identification of sensitive outcome measures. The management of patients in prodromal spinocerebellar ataxia type 2 may raise ethical dilemmas related to predictive diagnosis and early interventions, which impose new challenges to clinical and therapeutic research. © 2017 International Parkinson and Movement Disorder Society.


Vision and strategy for healthcare: Competence is a necessity.
Well-designed, evidence-based vision and strategy are fundamental requirements for successful healthcare. This paper reviews previous comprehensive healthcare strategies (Institute of Medicine statement, Value-based healthcare, and Triple aim strategy) published in peer-reviewed scientific journals, and presents an alternative proposal for healthcare vision and strategy. The proposed strategy includes, as prerequisites, competence of staff, healthcare leaders and leaders of health policy, and a well-functioning healthcare system. It is necessary to optimize the financing, reimbursement and incentives, organization, and regulation of the healthcare system. Strategic means to successful healthcare are to improve staff competence, implement current scientific evidence, document clinical and system-related issues, improve quality, and carry out benchmarking with peers. The vision for healthcare is to provide continuous improvement in services, in terms of accessibility, quality, fairness, effectiveness, safety and efficiency, to the patients (customers) and to the population. The new proposal shares many issues with strategies published previously. This paper sets out the prerequisites for the proposed strategy. A common vision, high levels of integrity, and a commitment to providing the best for patients and the population, are of fundamental importance. Competence at all levels of healthcare is a necessity.


Suppression of erythroid progenitor cells during malarial infection in Thai adults caused by serum inhibitor.
The bone marrows of 21 Thai adults infected with Plasmodium falciparum malaria were cultured for CFU-E and BFU-E by using AB serum, autologous serum (parasitaemia) and autologous serum (post-parasitaemia). Six patients had no complication and 15 patients had pulmonary, renal or haematologic complications. In the non-complicated cases, sera during parasitaemia did not suppress the post-parasitaemia CFU-E and BFU-E. Post parasitaemia, there was suppression of CFU-E by parasitaemia sera. In the complicated cases, the autologous sera during parasitaemia suppressed the growth of both CFU-E and BFU-E both during and after parasitaemia (P < 0.05). The post-parasitaemia sera had neither a suppressive nor a stimulating effect. In the complicated cases, the progenitor cells cultured from the bone marrow post-parasitaemia were fewer in number than those cultured from the bone marrow during parasitaemia using the same sera. Two possible mechanisms of suppression are postulated, namely the reduction of erythropoietin or the increased tumour necrosis factor during malarial infection. Further studies to clarify this are being carried out.


99mTc-TRODAT-1 SPECT/CT imaging as a complementary biomarker in the diagnosis of parkinsonian syndromes.
Parkinson's disease (PD) and Parkinson plus syndromes (PPS) are neurodegenerative movement disorders caused by loss of dopamine in the basal ganglia. The diagnosis of both PD and PPS is complex as it is made solely on the basis of clinical features, with no established imaging modality to aid in the diagnosis. Technetium-99m-labeled tropane derivative (Tc-TRODAT-1) binds to the dopamine transporters present in the presynaptic membrane of the dopaminergic nerve terminal. The aim of this prospective study was to investigate the potential usefulness of Tc-TRODAT-1 imaging in the diagnosis of PD and PPS. Fifty-eight patients with a clinical diagnosis of idiopathic PD or PPS were recruited. The severity of the disease was assessed using the Hoehn and Yahr scale. Patients in stage I and II were considered as cases of Early PD. Twenty-five apparently healthy volunteers served as controls. Brain single-photon emission computed tomography/computed tomography in all the participants was performed 3-4 h after an injection of Tc-TRODAT-1. Specific uptake ratios (SURs) of striatum were calculated for both the left and right striatum, and the values were compared between PD, PPS, and healthy volunteers. A significant lower uptake of tracer activity was found in either of the striatum in PD and PPS cases compared with the control group, which showed a symmetrical comma-shaped striatal uptake. This was also reflected in the SUR values, which were significantly higher in the control group in comparison with the PD and PPS patients (P<0.001). A significant difference was also found in the SUR values between the cases of early PD and control group (P<0.001).No significant difference was noted among the SUR values in different Hoehn and Yahr stages. For clinical practice, both the visual analysis and the quantitative parameters of Tc-TRODAT-1 single-photon emission computed tomography/computed tomography showed usefulness in distinguishing cases of PD and PPS from the healthy individuals.


Chronic Menthol Does Not Change Stoichiometry or Functional Plasma Membrane Levels of Mouse α3β4-Containing Nicotinic Acetylcholine Receptors.
Heteromeric α3β4 nicotinic acetylcholine (ACh) receptors (nAChRs) are pentameric ligand-gated cation channels that include at least two α3 and two β4 subunits. They have functions in peripheral tissue and peripheral and central nervous systems. We examined the effects of chronic treatment with menthol, a major flavor additive in tobacco cigarettes and electronic nicotine delivery systems, on mouse α3β4 nAChRs transiently transfected into neuroblastoma-2a cells. Chronic menthol treatment at 500 nM, near the estimated menthol concentration in the brain following cigarette smoking, altered neither the [ACh]-response relationship nor Zn2+ sensitivity of ACh-evoked currents, suggesting that menthol does not change α3β4 nAChR subunit stoichiometry. Chronic menthol treatment failed to change the current density (peak current amplitude/cell capacitance) of 100 μM ACh-evoked currents. Chronic menthol treatment accelerated desensitization of 100 and 200 μM ACh-evoked currents. Chronic nicotine treatment (250 μM) decreased ACh-induced currents, and we found no additional effect of including chronic menthol. These data contrast with previously reported, marked effects of chronic menthol on β2* nAChRs studied in the same expression system. Mechanistically, the data support the emerging interpretation that both chronic menthol and chronic nicotine act on nAChRs in the early exocytotic pathway, and that this pathway does not present a rate-limiting step to the export of α3β4 nAChRs; these nAChRs include endoplasmic reticulum (ER) export motifs but not ER retention motifs. Previous reports show that smoking mentholated cigarettes enhances tobacco addiction; but our results show that this effect is unlikely to arise via menthol actions on α3β4 nAChRs.


A three-step method of self-reflection using reflective journal writing.
Fiscal and financial constraints present a challenge for nurse educators to broaden the diversity and scope of teaching/learning methodologies. One method designed to promote autonomy and self-direction of nursing students is self-reflection combined with reflective journal writing. This paper describes a three-step process of self-reflection encompassing critical appraisal, peer group discussion and self-awareness. This process of self-reflection was initiated with one group of clinical nursing students. Using student and teacher feedback, implications for employing this teaching/learning strategy in clinical practice are suggested.


SmCOI1 affects anther dehiscence in a male-sterile Solanum melongena line.
Anther indehiscence is an important form of functional male sterility that can facilitate the production of hybrid seed; however, the molecular mechanisms of anther indehiscence-based male sterility have not been thoroughly explored in eggplant (Solanum melongena L.). Here, we used two-dimensional gel electrophoresis to compare the protein profiles in the anthers of normally developing (F142) and anther indehiscent (S16) S. melongena plants. Four differentially expressed proteins were identified using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry. Of these proteins, the transcript accumulation of the eggplant CORONATINE INSENSITIVE1 (SmCOI1) was significantly downregulated in S16 relative to F142. Phylogenetic analysis showed that SmCOI1 has high amino acid sequence similarity and clustered into the same subgroup as its homologs in other members of the Solanaceae. Subcellular localization analysis showed that SmCOI1 localized to the nucleus. Moreover, reverse-transcription quantitative PCR revealed that the jasmonic acid pathway genes SmJAZ1 and SmOPR3 are upregulated in F142 relative to S16. Protein-protein interaction studies identified a direct interaction between SmCOI1 and SmOPR3, but SmCOI1 failed to interact with SmJAZ1. These findings shed light on the regulatory mechanisms of anther dehiscence in eggplant.


Modeling tau pathology in human stem cell derived neurons.
Tau pathology is a defining characteristic of multiple neurodegenerative disorders including Alzheimer's disease (AD) and Frontotemporal Dementia (FTD) with tau pathology. There is strong evidence from genetics and experimental models to support a central role for tau dysfunction in neuronal death, suggesting tau is a promising therapeutic target for AD and FTD. However, the development of tau pathology can precede symptom onset by several years, so understanding the earliest molecular events in tauopathy is a priority area of research. Induced pluripotent stem cells (iPSC) derived from patients with genetic causes of tauopathy provide an opportunity to derive limitless numbers of human neurons with physiologically appropriate expression levels of mutated genes for in vitro studies into disease mechanisms. This review discusses the progress made to date using this approach and highlights some of the challenges and unanswered questions this technology has the potential to address.


Orotracheal intubation.
The advantages of endotracheal intubations are many: patency of airway is reasonably assured, secretions may be removed easily from the tracheobronchial tree, the airway is better protected against aspiration, ventilation can be regulated, and drugs can be administered. The procedure is the "gold standard" of airway control, and the oral route is the most common used. The patient is assessed for potential difficulties that may arise from abnormalities of face and mouth: large face, large tongue, hypoplastic mandible, prominent or diseased maxillary teeth or facial fractures; or abnormalities of the neck: potential or real cervical spine instability, swelling, or laryngeal trauma. A plan is devised, together with one or more alternatives, should unforeseen difficulties arise, and in adherence to the principle of never removing the compensations of a coping patient. The skill is learned and practiced so that a gentle facility is always present at times of greatest need.


A synthetic medium for Saccharomyces cerevisiae.
Growth of Saccharomyces cerevisiae strain Castelli 20 has been observed in a minimal synthetic medium using different glucose concentrations and without biotin, during the first 30 hrs of its development at 28 degrees C. The yeast's growth was observed spectrophotometrically at 675 nm reading the Optical Density every hour. The minimal medium of Sheperd et al. (1980), with glucose 15 g/L and biotin was modified: the vitamin was eliminated and the concentration of glucose was gradually reduced down to 0.5 g/L. At 5 g/L of glucose concentration and without vitamins the strain grew very well. From our results up to 30 hrs biotin has no influence on the yeast's growth. The medium would be useful to study Saccharomyces cerevisiae physiology during the early period of its development.


Inhibition of biofilm formation in Mycobacterium smegmatis by Parinari curatellifolia leaf extracts.
Tuberculosis (TB) is a serious public health problem worldwide. Mycobacterium tuberculosis (M. tuberculosis) grows as drug tolerant pellicles. Agents that inhibit biofilm formation in M. tuberculosis have the potential to reduce the disease treatment period and improve the quality of tuberculosis chemotherapy. Parinari curatellifolia (P. curatellifolia) leaf extracts are claimed to treat symptoms similar to tuberculosis in ethnomedicinal practices. Mycobacterium smegmatis (M. smegmatis) is a surrogate organism used in antimycobacterial drug discovery assays. In this study, the effect of the leaf extracts of P. curatellifolia on M. smegmatis growth and biofilm formation was investigated in order to determine the basis of its use in traditional medicinal use. Phytochemicals from P. curatellifolia leaves were prepared using a mixture of 50% dichloromethane (DCM): 50% methanol and by serial exhaustive extraction using different solvents of decreasing polarity. The solvents were used in the following order, hexane > dichloromethane > ethyl acetate > acetone >ethanol > methanol > water. The micro-broth dilution method was used as an antimycobacterial susceptibility test to screen for the extract that effectively inhibited M. smegmatis growth and biofilm formation. Biofilm quantification was performed by staining the biofilms with crystal violet and determining the amount of the stain using a spectrophotometer. In addition, the effects of combining the most active extract with kanamycin were also investigated. The minimum inhibitory concentrations (MIC) of the extracts were found to be 6.2 μg/ml for the acetone extract, 12.5 μg/ml for both the ethanol and the total extract and 50 μg/ml for both the methanol and ethyl acetate extracts. The ethanol extract, dichloromethane extract and water extract were the only extracts that effectively inhibited biofilm formation in M. smegmatis. Combining the ethanol extract with kanamycin enhanced the effect of the ethanol extract in terms of inhibition of biofilm formation. P. curatellifolia leaves contain phytochemicals that have the potential to be used both as antimycobacterial and anti-biofilm formation compounds.


Hs.137007 is a novel epigenetic marker hypermethylated and up-regulated in breast cancer.
This study was conducted to mine novel breast-specific unigenes and analyze their epigenetic regulation in breast cancer. Differential digital display and methylation analysis identified the Hs.137007 gene containing a Kelch domain as a candidate novel epigenetic marker. In 50 pairs of breast cancer tissues and nearby normal tissues the methylation level of the 14 CpG sites at the promoter region (-778 to -485) of the gene was higher in cancer tissues (72-93%) than in normal tissues (31-83%), with a high correlation rate (p<0.05). End-point RT-PCR and real-time RT-PCR revealed that Hs.137007 was up-regulated in cancer tissues. A clear relationship between high methylation levels and up-regulated expression was also observed in the cultured breast cell lines. The MCF7 (90-100%) and MDAMB468 (100%) cancer cell lines that showed higher methylation than the BT549 (20-90%) and 184B5 (10-100%) at the 14 CpGs also showed elevated gene expression. Taken together, these results indicate that the Hs.137007 gene is a novel gene specifically expressed in the breast that can be utilized as an epigenetic marker of breast cancer.


Pediatric Motor Vehicle-Pedestrian Accident: a Simulation Scenario for Emergency Medicine Trainees.
Simulation-based medical education is an evolving field that allows trainees to practice skills in a safe environment with no risk to patients. Recently, technology-enhanced simulation for emergency medicine learners has been shown to have favorable effects on learner knowledge and patient outcomes. In this report, a human patient simulator is used to familiarize emergency medicine trainees with the presentation and management of a pediatric motor vehicle-pedestrian accident is described.


Primary hemostasis in hemodilution--2) Infusion solutions.
Various infusion solutions are used during preoperative hemodilution or for substituting intraoperative blood loss. We examined the influence of saline 0.9%, albumin 5%, dextran 6% gelatin 3.5% and hydroxyethyl starch 6% (HES, mw 40,000 and 450,000) on primary hemostasis using the In-vitro bleeding test (IVBT) on composed blood samples of different hematocrit but constant platelet concentration. All infusion solutions tested caused longer bleeding times, depending on the volume used. Saline and albumin had the strongest effect, whereas high molecular weight HES or dextran showed hardly any impairment, if used in a quantity of less than 20% of the blood volume. Autologous plasma seems to be the best substitute in hemodiluted patients, at least if more than 20% of the blood volume has to be substituted by infusion solutions.


[Long-term efficacy and safety of simultaneous integrated boost radiotherapy in non-operative esophageal squamous cell carcinoma: a multicenter retrospective data analysis (3JECROG R-05)].
Objective: To analyze the survival benefits and treatment related toxic effects of simultaneous integrated boost intensity-modulated radiotherapy (SIB-RT) for non-operative esophageal squamous cell carcinoma patients. Methods: The data of 2 132 ESCC patients who were not suitable for surgery or rejected operation, and underwent radical radiotherapy from 2002 to 2016 in 10 hospitals of Jing-Jin-Ji Esophageal and Esophagogastric Cancer Radiotherapy Oncology Group (3JECROG) were analyzed. Among them, 518 (24.3%) cases underwent SIB (SIB group) and 1 614 (75.7%) cases did not receive SIB (No-SIB group). The two groups were matched with 1∶2 according to propensity score matching (PSM) method (caliper value=0.02). After PSM, 515 patients in SIB group and 977 patients in No-SIB group were enrolled. Prognosis and treatment related adverse effects of these two groups were compared and the independent prognostic factor were analyzed. Results: The median follow-up time was 61.7 months. Prior to PSM, the 1-, 3-, and 5-years overall survival (OS) rates of SIB group were 72.2%, 42.8%, 35.5%, while of No-SIB group were 74.3%, 41.4%, 31.9%, respectively (P=0.549). After PSM, the 1-, 3-, and 5-years OS rates of the two groups were 72.5%, 43.4%, 36.4% and 75.3%, 41.7%, 31.6%, respectively (P=0.690). The univariate survival analysis of samples after PSM showed that the lesion location, length, T stage, N stage, TNM stage, simultaneous chemoradiotherapy, gross tumor volume (GTV) and underwent SIB-RT or not were significantly associated with the prognosis of advanced esophageal carcinoma patients who underwent radical radiotherapy (P<0.05). Cox model multivariate regression analysis showed lesion location, TNM stage, GTV and simultaneous chemoradiotherapy were independent prognostic factors of advanced esophageal carcinoma patients who underwent radical radiotherapy (P<0.05). Stratified analysis showed that, in the patients whose GTV volume≤50 cm(3), the median survival time of SIB and No-SIB group was 34.7 and 30.3 months (P=0.155), respectively. In the patients whose GTV volume>50 cm(3), the median survival time of SIB and No-SIB group was 16.1 and 20.1 months (P=0.218). The incidence of radiation esophagitis and radiation pneumonitis above Grade 3 in SIB group were 4.3% and 2.5%, significantly lower than 13.1% and 11% of No-SIB group (P<0.001). Conclusions: The survival benefit of SIB-RT in patients with locally advanced esophageal carcinoma is not inferior to non-SIB-RT, but without more adverse reactions, and shortens the treatment time. SIB-RT can be used as one option of the radical radiotherapy for locally advanced esophageal cancer.


Poor Collateral Circulation Assessed by Multiphase Computed Tomographic Angiography Predicts Malignant Middle Cerebral Artery Evolution After Reperfusion Therapies.
Collateral circulation (CC) has been associated with recanalization, infarct volume, and clinical outcome in patients undergoing acute reperfusion therapies. However, its relationship with the development to malignant middle cerebral artery infarction (mMCAi) has not been evaluated. Our aim was to determine the impact of CC using multiphase computed tomographic angiography (during the acute stroke phase in the prediction of mMCAi. Patients with consecutive acute stroke with <4.5 hours who were evaluated for reperfusion therapies and presented with an M1-MCA or terminal internal carotid artery occlusion by CTA were included. CC was evaluated on 6 grades by multiphase CTA according to the University of Calgary CC Scale; CC status was defined as poor (grades, 0-3) or good (grades, 4-5). The mMCAi was defined according to clinical and radiological criteria. Recanalization was assessed with transcranial Doppler at 24 hours and final Thrombolysis in Brain Ischemia score≥2b in patients undergoing endovascular reperfusion treatment. Eighty-two patients were included. Mean age was 65.1±13.83 years, median baseline National Institutes of Health Stroke Scale score was 18 (interquartile range, 13-20), and 67.9% M1 and 32.1% terminal internal carotid artery occlusions. Fifty-three patients received endovascular reperfusion treatment. Fifteen patients developed mMCAi. In the univariate analysis, patients with mMCAi had lower CC scores (2.29 versus 3.71; P=0.001). Endovascular reperfusion treatment was associated with lower rate of mMCAi development than only intravenous reperfusion treatment (9.4% versus 29.6%; P=0.028). Patients with poor CC had higher risk of developing mMCAi (13% versus 2%; P=0.001). On the multivariate analysis adjusted by age, vessel occlusion, baseline National Institutes of Health Stroke Scale, and recanalization, the presence of poor CC by multiphase CTA was the only independent predictor of mMCAi (P=0.048; odds ratio, 9.72; 95% confidence interval, 1.387-92.53). CC assessment by multiphase CTA independently predicts malignant MCA infarction progression. In patients with persistent occlusion after reperfusion therapies, the presence of poor CC may improve the early mMCAi detection and management.


Global biogeography of reef fishes: a hierarchical quantitative delineation of regions.
Delineating regions is an important first step in understanding the evolution and biogeography of faunas. However, quantitative approaches are often limited at a global scale, particularly in the marine realm. Reef fishes are the most diversified group of marine fishes, and compared to most other phyla, their taxonomy and geographical distributions are relatively well known. Based on 169 checklists spread across all tropical oceans, the present work aims to quantitatively delineate biogeographical entities for reef fishes at a global scale. Four different classifications were used to account for uncertainty related to species identification and the quality of checklists. The four classifications delivered converging results, with biogeographical entities that can be hierarchically delineated into realms, regions and provinces. All classifications indicated that the Indo-Pacific has a weak internal structure, with a high similarity from east to west. In contrast, the Atlantic and the Eastern Tropical Pacific were more strongly structured, which may be related to the higher levels of endemism in these two realms. The "Coral Triangle", an area of the Indo-Pacific which contains the highest species diversity for reef fishes, was not clearly delineated by its species composition. Our results show a global concordance with recent works based upon endemism, environmental factors, expert knowledge, or their combination. Our quantitative delineation of biogeographical entities, however, tests the robustness of the results and yields easily replicated patterns. The similarity between our results and those from other phyla, such as corals, suggests that our approach may be of broad utility in describing and understanding global marine biodiversity patterns.


HIV type 1 inhibition by protein kinase C modulatory compounds.
The dichotomous effects of the protein kinase C (PKC) modulatory compounds 12-myristate 13-acetate (PMA), prostratin, and ingenol 3-angelate (I3A) on HIV-1 infection were investigated. PKC modulatory compounds were shown to be potent activators of cells latently infected with HIV-1 (I3A > prostratin). Conversely, PKC modulatory compounds inhibited infection of indicator cells (MAGI) with CXCR4-tropic HIV-1 (PMA > I3A > prostratin), and I3A also inhibited infection with CCR5-tropic virus (AD8-1). Pretreatment with the PKC inhibitors prior to treatment with either I3A or PMA resulted in increased infection, indicating inhibition is PKC mediated. Cell infections suggested that I3A rapidly inhibited the virus from infecting cells at an early point in infection. This observation was supported by the demonstration of inhibition at or before the synthesis of early reverse transcription products, and the inability of these compounds to block vesicular stomatitis virus (VSV) pseudotyped HIV-1 particles. As has already been shown with prostratin, treatment with I3A resulted in down-regulation of the CD4 receptor and CXCR4 coreceptor suggesting that this was a contributor to the infection inhibition. Intriguingly, 48 h pretreatment of unstimulated peripheral blood mononuclear cells (PBMC) prior to infection resulted in abrogation of virus production at concentrations where receptor/ coreceptor levels were not significantly reduced. This result hints at the possibility of inhibition by a PKC modulatory compound of an early pathway of viral entry in PBMC.


Characterization of interactions between the transcriptional repressor PhlF and its binding site at the phlA promoter in Pseudomonas fluorescens F113.
The phlACBD genes responsible for the biosynthesis of the antifungal metabolite 2,4-diacetylphloroglucinol (PHL) by the biocontrol strain Pseudomonas fluorescens F113 are regulated at the transcriptional level by the pathway-specific repressor PhlF. Strong evidence suggests that this regulation occurs mainly in the early logarithmic phase of growth. First, the expression of the phlF gene is relatively high between 3 and 13 h of growth and relatively low thereafter, with the phlACBD operon following an opposite expression profile. Second, the kinetics of PHL biosynthesis are specifically altered in the logarithmic phase in a P. fluorescens F113 phlF mutant. The phlA-phlF intergenic region presents a complex organization in that phlACBD is transcribed from a sigma(70) RNA polymerase-dependent promoter that is likely to overlap the promoter of the divergently transcribed phlF gene. The repression by PhlF is due to its interaction with an inverted repeated sequence, phO, located downstream of the phlA transcriptional start site. Cross-linking experiments indicate that PhlF can dimerize in solution, and thus PhlF may bind phO as a dimer or higher-order complex. Furthermore, it is now demonstrated that certain regulators of PHL synthesis act by modulating PhlF binding to phO. PHL, which has previously been shown to be an autoinducer of PHL biosynthesis, interacts with PhlF to destabilize the PhlF-phO complex. Conversely, the PhlF-phO complex is stabilized by the presence of salicylate, which has been shown to be an inhibitor of phlA expression.


Differentiation arrest and stromal cell-independent growth of murine erythroleukemia cells are associated with elevated expression of ets-related genes but not with mutation of p53.
The ELM erythroleukemia is novel in that long-term survival of leukemic cells in culture (ELM-D cells) is dependent on contact with a bone marrow-derived stromal feeder cell layer. However, a number of stroma-independent (ELM-I) mutants that vary in their ability to differentiate in vitro in response to erythropoietin and interleukin-3 have been derived. We have attempted to define the genetic changes responsible for these different phenotypes. At the p53 locus in the primary leukemic cells, one copy of the gene has been lost whereas the other contains an 18-bp depletion, implicating its mutation as an early step in the development of the leukemia. Changes in ets gene expression have also been found. The Fli-1 gene region is rearranged in the primary tumor because of the insertion of a retrovirus inserted upstream of one Fli-1 allele, but this does not result in Fli-1 gene activation in any of the ELM-D or ELM-I cell lines except one. It seems significant that this line is the only one to have lost the ability to differentiate in response to erythropoietin. In addition, up-regulation of erg is associated with stromal cell-independent growth, since all ELM-I mutants have moderate levels of erg mRNA, whereas only low or undetectable levels are found in primary leukemic cells in vivo or in ELM-D cells in vitro. This up-regulation of erg mRNA seems to be important for stromal cell-independent growth, since ELM-D cells show elevated expression of the erg gene after separation from stromal cells. This seems to be made permanent in ELM-I mutants, since they do not down-regulate erg mRNA when grown in contact with stromal cells. We therefore propose that ets family members regulate both the survival and differentiation of erythroid cells.


[Interaction among the relative risk factors of primary liver cancer in a case-control study].
This paper evaluates the combined effect of HBsAg, drinking water and aflatoxin B1 (AFB1) on the primary hepatocellular carcinoma (PHC) in a case-control study which was carried out in Fusui, Guangxi. We used relative risk model (from subadditive to supermultiplicative) to fit the relationship. The results showed that the combined effect of HBsAg infection and drinking pond-ditch water is nearly multiplicative, and the combined effect of HBsAg infection and AFB1 intake as well as drinking pond-ditch water and AFB1 intake were nearly additive.


[The prevention and treatment of recurrent urinary stress incontinence with the suspension according to Zoedler (author's transl)].
115 patients with severe urinary stress incontinence were operated according to the method of Zoedler with a nylon ribbon (31 cases) and a lyodura ribbon (84 cases). There were 56 primary operations and 59 operations for recurrence. In 13 patients the operation was combined with a fixation to the promentory according to Kuestner-Wagner. Continence was obtained in 90.3% of the cases at the end of one year in 84.2% of the cases at the end of three years and in 83% of the cases at the end of five years. The recurrence rate for the operations for recurrent stress incontinence was 20.6% the recurrence rate in the primary operations was 9.1%. The complications were described. The lyodura sling procedure is today primarily practiced as an operation for recurrent stress incontinence.


Adaptation of two primary human immunodeficiency virus type 1 isolates to growth in transformed T cell lines correlates with alterations in the responses of their envelope glycoproteins to soluble CD4.
Two sCD4-resistant, primary viruses (P-08 and P-17) were compared with two sCD4-sensitive, T cell line-adapted variants (C-08 and C-17) for their biochemical responses to sCD4. At 37 degrees C, neither primary virus shed gp120 within 8 hr at sCD4 concentrations of up to 500 nM, whereas C-08 and C-17 lost gp120 within minutes of addition of 5-10 nM sCD4. At 4 degrees C, however, P-17 and C-17 shed gp120 at similar rates in response to the same sCD4 concentration. Irrespective of the temperature, gp120 dissociation from both P-17 and C-17 was inhibited by CD4 MAbs 6H10 and 5A8, the latter of which blocks events subsequent to sCD4 binding. Binding of sCD4 to P-17 was greater at 4 degrees C than at 37 degrees C, whereas the converse was found for C-17. Consistent with this, P-17 was neutralized much more potently by sCD4 at 4 degrees C than at 37 degrees C, whereas C-17 was slightly more sensitive to sCD4 at 37 degrees C than at 4 degrees C. Resistance to neutralization by sCD4 is probably determined by kinetic parameters. We suggest that the acquisition of sCD4 neutralization sensitivity and the above biochemical responses to sCD4 are coincidental to the process by which some primary viruses adapt to growth in transformed T cells. Sequence data indicate that there are a limited number of amino acid differences between the Env glycoproteins of the primary viruses and their T cell line-adapted counterparts; the significance of the individual changes is under investigation, but both pairs of viruses have amino acid substitutions in a region of gp41 thought to contact gp120.