Altering the home care agency/client relationship: notice requirements.
Many older and disabled individuals regularly receive valuable services from home health agencies (HHAs). The unilateral termination or reduction of such services by an HHA may exert a significant impact on the life of a client who has come to depend on those services. The prerogatives of Medicare-certified HHAs to terminate their relationships with clients are constrained today not only by contract and tort law principles, but also by federal statutes and regulations establishing Conditions of Participation, including provisions concerning clients' rights. A recent important federal judicial decision interpreted and expanded the legal responsibilities of HHAs to provide formal notice to their Medicare clients before terminating or reducing home health care services to those clients, regardless of the reason for ending or altering the relationship. This article critically discusses the background, holding, and practice implications of the 2004 Lutwin v. Thomson decision, which imposes these notice requirements on HHAs.


Non-Hodgkin's lymphoma in children: an analysis of 122 cases from Argentina.
One hundred twenty two children with non-Hodgkin's lymphoma were studied from January 1966 to December 1975. The first group (1966-1972) did not receive an uniform treatment. The second group (1973-1975) entered in a G.A.T.L.A. protocol consisting of: vincristine-prednisone plus surgery and/or radiotherapy as induction treatment, craniocervical radiotherapy and intrathecal methotrexate as CNS preventive treatment and anti-leukemia (6-mercaptopurine, methotrexate and vincristine-prednisone pulses) or anti-lymphoma (COPP) treatment as maintenance, in a randomized trial. Comparison of survival of the two groups are as follows: series 1966-1972, 22% and 20% at 12 and 24 months of evolution, respectively, and series 1973-1975, 33% and 26% at 12 and 24 months, respectively. After 2 years of complete remission we have not seen any relapse. We conclude that 1) this disease is highly malignant and must be treated with more intensive chemotherapeutic treatment, and 2) there is no difference between antileukemia or anti-lymphoma maintenance treatment, as yet.


Oestrogen changed cardiomyocyte contraction and beta-adrenoceptor expression in rat hearts subjected to ischaemia-reperfusion.
Women with functional ovaries have a lower cardiovascular risk than men and postmenopausal women. However, oestrogen replacement therapy remains controversial. This study examined the effect of ovarian hormone deficiency and oestrogen replacement on ventricular myocyte contractile function and expression of beta-adrenoceptors (beta-ARs). Female Sprague-Dawley rats were subjected to bilateral ovariectomy (OVX) or sham operation (Sham). A subgroup of OVX rats received oestrogen (E2) replacement (40 microg kg(-1) day(-1)) for 4 weeks. Cardiomyocyte shortening was evaluated in basal conditions and in the presence of isoprenaline (ISO). The expression of beta-ARs was assessed by Western blotting. The presence of lactate dehydrogenase (LDH) activity in the coronary effluent was determined. Ovariectomy promoted body weight gain associated with reduced serum E2 and uterine weight, all of which were abolished by treatment with E2. Ovariectomy increased the amplitude of both basal and ISO-stimulated contractions, increased LDH release, upregulated beta1-AR expression and downregulated beta2-AR expression, all of which were restored by treatment with E2. A beta1-AR antagonist, CGP20712A, but not a beta2-AR antagonist, ICI118,551, significantly decreased the amplitude of ventricular myocyte shortening. Oestrogen decreased cardiomyocyte contraction and the expression of beta1-AR, and increased expression of beta2-AR, and all these effects were abolished by the E2 receptor antagonist, ICI182,780. These data suggest that oestrogen plays a cardioprotective role in female rat hearts subjected to ischaemia-reperfusion injury, and the effects of oestrogen are associated with decreased cardiomyocyte contraction and expression of beta1-AR, and increased expression of beta2-AR.


Phosphoprotein stability in clinical tissue and its relevance for reverse phase protein microarray technology.
Phosphorylated proteins reflect the activity of specific cell signaling nodes in biological kinase protein networks. Cell signaling pathways can be either activated or deactivated depending on the phosphorylation state of the constituent proteins. The state of these kinase pathways reflects the in vivo activity of the cells and tissue at any given point in time. As such, cell signaling pathway information can be extrapolated to infer which phosphorylated proteins/pathways are driving an individual tumor's growth. Reverse phase protein microarrays (RPMAs) are a sensitive and precise platform that can be applied to the quantitative measurement of hundreds of phosphorylated signal proteins from a small sample of tissue. Pre-analytical variability originating from tissue procurement and preservation may cause significant variability and bias in downstream molecular analysis. Depending on the ex vivo delay time in tissue processing, and the manner of tissue handling, protein biomarkers such as signal pathway phosphoproteins will be elevated or suppressed in a manner that does not represent the biomarker levels at the time of excision. Consequently, assessment of the state of these kinase networks requires stabilization, or preservation, of the phosphoproteins immediately post-tissue procurement. We have employed RPMA analysis of phosphoproteins to study the factors influencing stability of phosphoproteins in tissue following procurement. Based on this analysis we have established tissue procurement guidelines for clinical research with an emphasis on quantifying phosphoproteins by RPMA.


Volatile fatty acid concentrations in the digestive tract of the West Indian manatee, Trichechus manatus.
1. Digesta samples were collected from five West Indian manatees, Trichechus manatus, for volatile fatty acid (VFA) analysis. 2. Mean total VFA concentrations were low in the stomach and duodenum (18.6 and 12.3 mM/l, respectively). Mean VFA concentrations were considerably higher in the cecum and colon (220.6 and 307.3 mM/l, respectively). 3. The relative proportions of the individual VFA's shifted from predominantly acetic acid in the foregut to a mixture of acetic, butyric and propionic acids in the hindgut. 4. The VFA concentrations in the manatee are similar to those in the dugong, Dugong dugon, and the green sea turtle, Chelonia mydas. 5. The mean total VFA concentrations indicate that the cecum and colon of the manatee are sites of microbial cellulose fermentation. The contribution of VFAs to the manatees' total energy requirements could not be calculated, but it is probably considerable.


Possible participation of receptor for advanced glycation end products (RAGE) in the origin of cancer stem cells in diabetic patients with colon cancer.
The association between diabetes and the associated increased risk of several solid malignancies has been the subject of investigation for many years, while potential biologic links between the two diseases are incompletely understood. The receptor for advanced glycation end-products (RAGE) signal transduction may represent a focal point in their respective contributions to malignant transformation associated diabetes. While the physiopathology of RAGE axis in promoting malignancies cannot be explained completely by the available mechanism as perpetuating inflammation at tumor microenvironment. In addition, experimental researches revealed a crucial role for upstreams of RAGE signaling pathway in maintaining the stemness properties and tumorigenicity of cancer stem cells. Hence, we hypothesized that RAGE inducing cancer stem cells may be a key determinant in the origin and progression of colon malignant tumors concomitant diabetes. Such an opinion not only bands together the seemingly disparate various complications in diabetes and colon cancers, but also has future implications for risk assessment and biopharmaceutical treatment.


Lysobacter telluris sp. nov., isolated from Korean rhizosphere soil.
A Gram-stain-negative, aerobic, non-motile, non-spore-forming light-yellow-coloured rod-shaped bacterial strain, designated YJ15T, was isolated from soil at Bigeum island in Korea. Growth was observed at 10-37 °C (optimum, 28 °C), at pH 6.0-7.5 (optimum, pH 7.0) and in the absence of NaCl. Based on 16S rRNA gene sequence analysis, strain YJ15T was closely related to 'Lysobacter tongrenensis' YS037T (97.8%), Lysobacter pocheonensis Gsoil193T (96.5%) and Lysobacter daecheongensis Dae08T (95.8%) and phylogenetically grouped together with 'Lysobacter tongrenensis' YS037T, Lysobacter dokdonensis DS-58T and Lysobacter pocheonensis Gsoil 193T. The DNA-DNA relatedness between strain YJ15T and 'Lysobacter tongrenensis' KCTC 52206T was 12% and the phylogenomic analysis based on the whole genome sequence demonstrated that strain YJ20T formed a distinct phyletic line with Lysobacterlter dokdonensis DS-58T showing average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values of 76.3 and 21.3%, respectively. The predominant ubiquinone was identified as Q-8, and polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and two unidentified aminolipids. The major fatty acids were iso-C17:1 ω9c, iso-C15:0, iso-C16:0 and iso-C17:0. The genomic DNA G + C content was 68.2 mol %. On the basis of phenotypic, chemotaxonomic properties and phylogenetic analyses in this study, strain YJ15T is considered to represent a novel species of the genus Lysobacter, for which the name Lysobacter telluris sp. nov. is proposed. The type strain is YJ15T (= KACC 19552T = NBRC 113197T).


The seasonal dynamics of leaf resin, nitrogen, and herbivore damage in Eriodictyon californicum and their parallels in Diplacus aurantiacus.
The chaparral shrub Eriodictyon californicum produces a flavonoid leaf resin with a chemically similar composition to that previously reported for the sympatric shrub Diplacus aurantiacus. We determined the phenology, resin content, and nitrogen content of Eriodictyon leaves and the leaf area lost to herbivores. Nitrogen content and resin content were both negatively correlated with leaf age at each sampling date, but nitrogen decreased during the growing season while resin increased. The fraction of leaf area lost during the growing season averaged less than 7% and was highest on the oldest leaves. The seasonal pattern of resin production in Eriodictyon corresponds to that in Diplacus, indicating that the similar environments of Eriodyctyon and Diplacus have led to convergent leaf resins. This convergence in these two plants has implications for chemicals of similar function in other chaparral shrubs.


Disruption of mindin exacerbates cardiac hypertrophy and fibrosis.
Cardiac hypertrophy is a response of the myocardium to increased workload and is characterised by an increase of myocardial mass and an accumulation of extracellular matrix (ECM). As an ECM protein, an integrin ligand, and an angiogenesis inhibitor, all of which are key players in cardiac hypertrophy, mindin is an attractive target for therapeutic intervention to treat or prevent cardiac hypertrophy and heart failure. In this study, we investigated the role of mindin in cardiac hypertrophy using littermate Mindin knockout (Mindin ( -/- )) and wild-type (WT) mice. Cardiac hypertrophy was induced by aortic banding (AB) or angiotensin II (Ang II) infusion in Mindin ( -/- ) and WT mice. The extent of cardiac hypertrophy was quantitated by echocardiography and by pathological and molecular analyses of heart samples. Mindin ( -/- ) mice were more susceptible to cardiac hypertrophy and fibrosis in response to AB or Ang II stimulation than wild type. Cardiac function was also markedly exacerbated during both systole and diastole in Mindin ( -/- ) mice in response to hypertrophic stimuli. Western blot assays further showed that the activation of AKT/glycogen synthase kinase 3β (GSK3β) signalling in response to hypertrophic stimuli was significantly increased in Mindin ( -/- ) mice. Moreover, blocking AKT/GSK3β signalling with a pharmacological AKT inhibitor reversed cardiac abnormalities in Mindin ( -/- ) mice. Our data show that mindin, as an intrinsic cardioprotective factor, prevents maladaptive remodelling and the transition to heart failure by blocking AKT/GSK3β signalling.


Quantitative evaluation of gait ataxia by accelerometers.
An appropriate biomarker for spinocerebellar degeneration (SCD) has not been identified. Here, we performed gait analysis on patients with pure cerebellar type SCD and assessed whether the obtained data could be used as a neurophysiological biomarker for cerebellar ataxia. We analyzed 25 SCD patients, 25 patients with Parkinson's disease as a disease control, and 25 healthy control individuals. Acceleration signals during 6 min of walking and 1 min of standing were measured by two sets of triaxial accelerometers that were secured with a fixation vest to the middle of the lower and upper back of each subject. We extracted two gait parameters, the average and the coefficient of variation of motion trajectory amplitude, from each acceleration component. Then, each component was analyzed by correlation with the Scale for the Assessment and Rating of Ataxia (SARA) and the Berg Balance Scale (BBS). Compared with the gait control of healthy subjects and concerning correlation with severity and disease specificity, our results suggest that the average amplitude of medial-lateral (upper back) of straight gait is a physiological biomarker for cerebellar ataxia. Our results suggest that gait analysis is a quantitative and concise evaluation scale for the severity of cerebellar ataxia.


Radiation dermatitis assessment tools used in breast cancer: A systematic review of measurement properties.
This review aimed to assess the quality and efficacy of tools currently used in breast cancer patients to score radiation dermatitis (RD), a common debilitating side effect of radiotherapy (RT). A search was conducted through Ovid Medline, Embase, and Cochrane Central Register of Controlled Trials databases on 14 February 2020. English articles that evaluated an instrument's use in assessing RD among breast cancer patients receiving external beam RT were included. Studies that reported on the reliability, validity, or concordance of items between assessment tools were included in accordance with the Consensus-Based Standards for the Selection of Health Measurement Instruments (COSMIN) criteria. Twelve studies were included in this review, with a total of 13 skin toxicity assessment tools discussed. Tools that assessed clinician-reported outcomes (CROs) mostly reported moderate correlation with biophysical parameter (BP) measurements and low correlation with patient-reported outcomes (PROs). Traditionally used CRO scoring tools demonstrated moderate inter-rater reliability between clinicians, likely due to the subjective nature of items on the grading scales. Most commonly used tools were found to be either insufficient or indeterminate in their measurement properties. Current standardized tools that measure CROs are subject to clinician interpretation and fail to represent the patient experience. Tools designed to assess PROs are promising in their assessments of the impact of RT on patient quality of life; however, most PRO tools are generic to all skin conditions and require further validation for use in breast cancer. Among tools that measure CROs, PROs, and BPs, there is insufficient evidence on their measurement properties to establish a "gold standard" for the assessment of RD in breast cancer patients.


Antigen-specific electrophoretic cell separation (ASECS): isolation by human T and B lymphocyte subpopulations by free-flow electrophoresis after reaction with antibodies.
The electrophoretic mobility of human lymphocytes can be reduced by incubation with surface antigen specific antibodies under non-capping conditions. This renders subpopulations of human peripheral blood lymphocytes accessible to separation by free-flow electrophoresis. After reaction of lymphocyte preparations with anti-IgM antibody and a fluorescent second antibody, B lymphocytes showed a considerable shift in position in preparative cell electrophoresis and could be separated with high yield, purity and vitality. Similarly, a T cell subpopulation reactive with the monoclonal antibody T811 could be isolated, even though only small amounts of this antibody were bound, by using a double-sandwich method. Non-specific antibody uptake via Fc-receptors did not contribute to the observed shift of antibody-labelled cells to lower electrophoretic mobility. Flow cytometric analysis showed that cells were separated according to their antigen density. Thus cell electrophoresis can be used to separate antibody-labelled cells. With a flow rate of 100,000 cells/sec this method has a much higher separation capacity than fluorescence-activated cell sorting. The described method should be applicable to the separation of a wide range of cell populations for which specific antibodies are available.


Changes in hemostasis during pediatric heart surgery: impact of a biocompatible heparin-coated perfusion system.
This study describes the response in hemostasis during open-heart surgery with cardiopulmonary bypass (CPB) in children (<== 10 kg) and tests the hypothesis that the use of a biocompatible perfusion system, in comparison with a conventional system, causes less hemostatic activation. Prospective, randomized, controlled clinical study. Forty consecutive children <== 10 kg were included and divided into two groups: group bioc. (n = 19) treated with a fully heparin-coated system, centrifugal pump, and a closed circuit, and group conv. (n = 21) treated with an uncoated system, roller pump, and a hard shell venous reservoir. Concentrations of plasma thrombin-antithrombin (TAT), D-dimer, tissue plasminogen activator antigen (t-PA ag), and the complex consisting of tissue plasminogen activator and its inhibitor plasminogen activator inhibitor-1 (t-PA-PAI-1) were measured. The biochemical variables measured increased significantly in both groups during the study period. There was less activation of fibrinolysis during cardiopulmonary bypass (t-PA ag: p = 0.009) in patients treated with the biocompatible perfusion system than in patients treated with the conventional system. A trend in favor of the biocompatible system based on the D-dimer and TAT data (p = 0.07 for both measurements) was observed but no significant intergroup differences regarding these variables or t-PA-PAI-1 were found. Open-heart surgery with cardiopulmonary bypass in children (<== 10 kg) causes transient activation of the coagulation and fibrinolytic systems. This study demonstrates that the use of a biocompatible perfusion system results in a lower extent of activation of fibrinolysis during CPB than the use of a conventional system.


Relationship between shear modulus and passive tension of the posterior shoulder capsule using ultrasound shear wave elastography: A cadaveric study.
Although shear wave elastography (SWE) has been used to indirectly measure passive tension in muscle tissues, it is unknown whether SWE can adequately evaluate passive tension in capsule tissues. This study investigated the relationship between the shear modulus and passive tension in the posterior shoulder capsule using SWE. Ten posterior middle and ten posterior inferior shoulder capsules were dissected from ten fresh-frozen cadavers; humeral head-capsule-glenoid specimens were created from each capsule. The humeral head and glenoid were immobilized with clamps in a custom-built device. Loads (0-400 g, in 25-g increments) were applied to each capsule via a pulley system; elasticity was simultaneously measured using SWE. The elasticity-load relationship of each tested capsule was analyzed by fitting a least-squares regression line to the data. Elasticity change due to creep or hysteresis effects was evaluated by comparing the elastic modulus for a 100-g load during and after the stepwise application of the loads. The observed relationship between the shear modulus and passive capsule tension was highly linear for all twenty tested capsules (p < 0.01). The mean coefficient of determination was 0.882 ± 0.075 and 0.901 ± 0.050 for the posterior middle and posterior inferior capsules, respectively. There was no difference in the shear modulus between the two 100-g load assessments for both the posterior middle (p = 0.205) and posterior inferior capsules (p = 0.161). Thus, SWE is a valid and useful method for indirectly evaluating the change in the passive tension under loading in specific posterior shoulder capsule.


Contributions of protein and milled chitin extracted from domestic cricket powder to emulsion stabilization.
Interfacial and emulsifying properties of fractionated cricket powder were assessed to identify whether emulsification properties originate from protein or chitin particles. Fractions extracted in alkaline water, containing high protein and mineral contents, increased the surface pressure of heptane-water interfaces with near-saturation equilibrium surface pressure of 31 mN/m. Dynamic surface pressure profiles indicated adsorption of protein clusters to the interface. Emulsification capacity of protein fraction was 50% greater than that of the source cricket flour, although oil-in-water emulsions prepared with 1-2% (w/w) protein fraction formed a cream layer within one day of storage. Emulsified layers persisted for up to 20 days, and light scattering measurements described a stable population with surface-volume-mean diameter of approximately 3 μm. Chitin-rich fractions milled to a particle size of 0.5-200 μm contributed negligible surface pressure, and its emulsification capacity was 5% of the value for the source cricket flour. Emulsions prepared with chitin-rich fractions coexisted with an unstable precipitate layer comprising 60% of the added solid, which was attributed to larger particles with poor emulsifying capability. Stable chitin-stabilized emulsion phases were resistant to creaming, yet volume-mean droplet diameter surpassed 50 μm within 24 h of storage. Both protein and chitin fractions have emulsifying capabilities but would require further processing or secondary additives to achieve desirable storage stability.


Preliminary investigations into the involvement of the intestinal microflora in CNS toxicity induced by 1,3-dinitrobenzene in male F-344 rats.
Administration of a single oral dose of 20 mg/kg of 1,3-dinitrobenzene caused ataxia in germ-free male F-344 rats but not in conventional rats. Repeated oral dosing of 20 mg/kg, 1,3-DNB was required to cause ataxia in conventional rats. Considerable differences were observed between the uptake, tissue distribution and excretion of DNB in germ-free and conventional rats.


Microcalorimetry: a novel method for detection of microbial contamination in platelet products.
Measuring heat from replicating microorganisms in culture may be a rapid, accurate, and simple screening method for platelets (PLTs). Microcalorimetry for detection of microorganisms in in vitro contaminated PLT products was evaluated. Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus sanguinis, Escherichia coli, Propionibacterium acnes, and Candida albicans were inoculated in single-donor apheresis PLTs to achieve target concentrations of 10(5), 10(3), 10, or 1 colony-forming units (CFU) per mL of PLTs. Contaminated PLTs in growth medium were incubated at 37 degrees C for 5 days in a calorimeter. Positivity was defined as heat flow of at least 10 microW above the lowest value of the power-time curve. With microcalorimetry, inocula of 10 CFUs per mL PLTs could be detected with the following detection times: S. epidermidis (31.65 hr), S. aureus (24.24 hr), S. sanguinis (7.82 hr), E. coli (7.53 hr), P. acnes (73.57 hr), and C. albicans (43.77 hr). The detection time was less than 4 hr at 10(5) CFUs per mL PLTs for S. aureus, S. sanguinis, and E. coli. Noncontaminated PLTs remained negative. The total heat ranged from 2.8 (S. sanguinis) to 8.3 J (E. coli). The shape of the power-time curve was species-specific and independent from the initial concentration of microorganisms. The detection limit of microcalorimetry was 1 to 10 CFUs per mL PLTs. Microcalorimetry is a promising novel method for detection of contaminated PLTs. Applying this method to all PLT products could reduce the frequency of transfusion-related sepsis and prolong the shelf life of PLTs.


Protective effect of egg yolk phosvitin against ultraviolet- light-induced lipid peroxidation in the presence of iron ions.
It is known that nonheme iron accumulates and free radicals are generated in skin exposed to ultraviolet (UV) light. Iron ions have a role in skin photodamage by participating in the formation of reactive oxygen species. In this study, we evaluated the effect of egg yolk phosvitin on UV-light-induced oxidative stress. Mouse dorsal skin homogenate was exposed to UVA light in the presence or absence of ferric nitrilotriacetate (FeNTA). Lipid peroxidation was determined by measuring thiobarbituric acid-reactive substances (TBARS). The TBARS concentration increased with increasing FeNTA concentration and UV-light-exposure time. In the presence of FeNTA, phosvitin more effectively inhibited in vitro lipid peroxidation than did bovine serum albumin. According to results of electron spin resonance studies using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) as a spin trapping agent, phosvitin suppressed the formation of hydroxyl radicals. These results suggest that UV-light-induced oxidative stress can be reduced by phosvitin.


Spectrophotometric monitoring of nimesulide photodegradation by a combined hard-soft multivariate curve resolution-alternative least square method.
Nimesulide, a Cox-2 inhibitor anti-inflammatory drug, is a light sensitive compound and its biological activity is decreased upon photodegradation. The photodegradation kinetic of nimesulide was investigated spectrophotometrically using multivariate curve resolution analysis to overcome spectral overlapping of reactant and degradation products. The absorbance spectra of the nimesulide methanolic solution, exposed to daily sunlight, were recorded at different times. Three absorbing chemical species, coexisted in the reaction system, were detected by application of factor analysis to the absorbance data matrix. The soft-modeling multivariate curve resolution-alternative least square (MCR-ALS) analysis of the evolutionary absorbance data revealed that nimesulide undergoes photodegradation through a two-step consecutive manner, where both steps obey first-order kinetic. By application of the kinetic hard model constraint to the MCR-ALS analysis, an excellent agreement was obtained between the fitted concentration profiles and those obtained by soft method. The first-order rate constants of the first and second degradation products were calculated as 0.052 (+/-0.007) and 0.009 (+/-0.001)h(-1), respectively. Finally, the pure spectra of the resolved chemical species were calculated, where that of nimesulide was the same as that obtained experimentally.


Nutritional status and its associated factors among commercial female sex workers in Hawassa city, south Ethiopia.
Commercial female sex workers (CFSWs) regularly or occasionally trade sex for money. Sex work is widespread, mainly in urban areas of Ethiopia. The study on the nutritional status of CFSWs is unavailable in Ethiopia, and data are also scanty at the global level. This study aims to assess nutritional status and associated factors among CFSWs in Hawassa city, Ethiopia. This cross-sectional study used facility-based mixed data collection methods (qualitative and quantitative). The study was conducted in three key population clinics in Hawassa city. A total of 297 CFSWs were randomly selected for the 'quantitative survey,' and twelve (n = 12) purposively selected participants were involved in the 'qualitative study.' Body mass index/BMI (Kg/m2) was used in assessing the nutritional status of CFSWs. Statistical software packages were used for analyzing both quantitative and qualitative data. Significant variables (P<0.05) in bivariate analysis (Chi-square test) were incorporated in the multivariable analyses. Multinomial logistic regression (MNLR) was employed where dependable variable like 'normal BMI' (18.5-24.9 kg/m2) was set as a reference category and simultaneously compared with 'underweight' (<18.5 BMI) and 'overweight/obesity' (≥ 25 BMI) categories. Thus, two models, namely the underweight model (model-1: Underweight versus normal BMI) and the overweight/obesity model (model-2: overweight/obesity versus normal), were generated. The prevalence of underweight and overweight/obesity among CFSWs in Hawassa city were respectively 14.1% and 16.8%. Living alone (Adjusted odds ratio/AOR = 0.18), chewed Khat regularly (AOR = 0.23), used drugs regularly (AOR = 10.57), used drugs in exchange of sex (AOR = 4.97), and HIV positive status (AOR = 21.64) were significantly (P < 0.05) associated with underweight (model-1). In the overweight/obesity model-2, having jobs other than sex work (AOR = 0.11), higher daily mean income (AOR = 3.02), being hotel/home-based CFSWs (AOR = 12.35), and presence of any chronic illness (AOR = 5.15) were significant (P < 0.05) predictors of overweight/obesity. From the 'qualitative part' of this study, it was also revealed that 'lack of food and money' was the main influencing factor among CFSWs to enter into the sex business. Commercial female sex workers in this study faced a double burden of malnutrition. Multiple factors influenced their nutritional status. Substance abuse and HIV-positivity are the most significant predictors of being underweight and having higher income, being hotel/home-based CFSWs, and suffering from any chronic illness are associated with overweight/obesity. Government and other partners should be essential in providing comprehensive programs focusing on sexual, reproductive health, and nutrition education. Steps should be taken to improve their socioeconomic status and strengthen those good initiatives at key population clinics and other health facilities.


Aspirin increases ferroportin 1 expression by inhibiting hepcidin via the JAK/STAT3 pathway in interleukin 6-treated PC-12 cells.
To understand the potential mechanisms involved in the beneficial effects of aspirin (ASA) in mood disorders, Alzheimer's (AD) and Parkinson's disease (PD), we investigated the effects of ASA on the expression of iron transport proteins transferrin receptor 1 (TfR1), ferroportin 1 (Fpn1), and iron storage protein ferritin light chain (Ft-L) in interleukin-6 (IL-6)-treated PC-12 cells. We demonstrated that IL-6 alone could induce a severe decline in Fpn1 expression and cell viability, and an increase in Ft-L protein, while ASA could markedly diminish the effects of IL-6 on these parameters. We also found that IL-6 significantly increased hepcidin expression and janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) phosphorylation, while ASA also observably suppressed these IL-6-induced effects. The data imply that ASA increases Fpn1 expression by inhibiting hepcidin expression via the IL-6/JAK/STAT3 pathway and show that the reduced content of Ft-L is due to the increased Fpn1 and subsequent iron release in the cells. The reduction of iron in neuronal cells by the increased expression of Fpn1 might be partly associated with the beneficial effects of ASA on mood disorders, AD and PD.


A novel approach to identify the mechanism of miR-145-5p toxicity to podocytes based on the essential genes targeting analysis.
MicroRNAs (miRNAs) are emerging as effective therapeutic agents. When testing whether miR-145-5p could alleviate kidney injury, we unexpectedly found that extracellular vesicles loaded with miR-145-5p induced proteinuria and podocyte foot process effacement in normal control mice. To explore the mechanism of miR-145-5p's toxicity to podocytes, we hypothesized that miR-145-5p could enter podocytes and inhibit genes essential for podocytes. We demonstrated that systemically administered miRNA can enter podocytes. Next, we predicted 611 podocyte essential genes based on single-cell RNA sequencing (RNA-seq) and found that 32 of them are predicted to be targeted by miR-145-5p. Functional annotation of the 32 podocyte essential genes revealed small GTPase-mediated signal transduction as the top pathway. We experimentally validated that miR-145-5p targeted Arhgap24 and Srgap1, the essential regulators of the Rho family of small GTPases, increased the activity of Rac1 and Cdc42, and reduced RhoA activity, accompanied by cellular injury, in podocytes. These results explain how miR-145-5p has deleterious effect on podocytes. Most importantly, our study provides a novel approach to investigate how a miRNA affects a given cell type, allowing not only identification of the molecular mechanism underlying an observed side effect of a miRNA drug but also prediction of miRNA drug toxicity on various cell types.


[Nontuberculous mycobacterial infection after tattooing caused by Mycobacterium chelonae].
We report the case of a healthy young man who presented to our clinic with itchy skin lesions in the area of a tattoo on the back of the left hand. Bioptic and cultural confirmation of the pathogens led to the diagnosis of Mycobacterium chelonae infection. We initiated antibiotic therapy using azithromycin and linezolid with good response. Our case underlines that besides allergic skin reactions, infections as a complication after tattooing should also be included in the differential diagnosis.


CD4+ T-lymphocyte enumeration with a flow-rate based method in three flow cytometers with different years in service.
CD4(+) T-lymphocyte count remains the most important surrogate marker for management of HIV patients in resource-poor countries. The standard single-platform (SP) bead-based flow cytometric method for CD4(+) testing is expensive; more affordable methods are needed. We evaluated the SP flow-rate-based calibration method for determining CD4(+) counts, using three flow cytometers of varying ages. CD4(+) counts from 103 HIV-1 infected Thai patients were determined using a SP flow-rate method in flow cytometers with 2, 12, and 16 years of service. Results were compared to the bead-based method. Correlation and agreement were analyzed using linear regression and Bland-Altman analysis. Counts obtained from the flow-rate approach in each flow cytometer showed strong correlation with the bead-based method (R(2) = 0.97, 0.97, and 0.96 for the 2-, 12-, and 16-year-old flow cytometers). The mean biases for the flow-rate approach compared with the bead-based method were +32.4 cells/microL (limit of agreement (LOA): -83.0 to +147.8 cells/microL), -28.8 cells/microL (LOA: -131.6 to +74.1 cells/microL), and -27.0 cells/microL (LOA: -149.4 to +95.4 cells/microL). The flow-rate approach is reliable for determining CD4(+) counts. Results do not vary by age of flow cytometer. This approach provides a cost-effective alternative for HIV patient monitoring in resource-poor settings.


Antibiotic use in US hospitals: quantification, quality measures and stewardship.
A majority of patients hospitalized in the US hospitals receive an antibiotic during their hospitalization. Furthermore, up to half of antibiotics prescribed in hospitals are inappropriate. In the setting of continued emergence of antibiotic-resistant pathogens and a limited pipeline of new antimicrobials, attention to optimizing antibiotic use in healthcare settings is essential. We review the measures of antibiotic consumption in the USA, the evolving metrics for comparing antibiotic use (known as benchmarking), trends in antibiotic use, the structure and outcome measures of Antimicrobial Stewardship Programs and interventions to optimize antimicrobial use.


Epithelial mesenchymal transition traits in human breast cancer cell lines.
Epithelial mesenchymal transition (EMT) has long been associated with breast cancer cell invasiveness and evidence of EMT processes in clinical samples is growing rapidly. Genome-wide transcriptional profiling of increasingly larger numbers of human breast cancer (HBC) cell lines have confirmed the existence of a subgroup of cell lines (termed Basal B/Mesenchymal) with enhanced invasive properties and a predominantly mesenchymal gene expression signature, distinct from subgroups with predominantly luminal (termed Luminal) or mixed basal/luminal (termed Basal A) features (Neve et al Cancer Cell 2006). Studies providing molecular and cellular analyses of EMT features in these cell lines are summarised, and the expression levels of EMT-associated factors in these cell lines are analysed. Recent clinical studies supporting the presence of EMT-like changes in vivo are summarised. Human breast cancer cell lines with mesenchymal properties continue to hold out the promise of directing us towards key mechanisms at play in the metastatic dissemination of breast cancer.


C-terminal regulatory domain of the ε subunit of Fo F1 ATP synthase enhances the ATP-dependent H+ pumping that is involved in the maintenance of cellular membrane potential in Bacillus subtilis.
The ε subunit of Fo F1 -ATPase/synthase (Fo F1 ) plays a crucial role in regulating Fo F1 activity. To understand the physiological significance of the ε subunit-mediated regulation of Fo F1 in Bacillus subtilis, we constructed and characterized a mutant harboring a deletion in the C-terminal regulatory domain of the ε subunit (ε∆C ). Analyses using inverted membrane vesicles revealed that the ε∆C mutation decreased ATPase activity and the ATP-dependent H+ -pumping activity of Fo F1 . To enhance the effects of ε∆C mutation, this mutation was introduced into a ∆rrn8 strain harboring only two of the 10 rrn (rRNA) operons (∆rrn8 ε∆C mutant strain). Interestingly, growth of the ∆rrn8 ε∆C mutant stalled at late-exponential phase. During the stalled growth phase, the membrane potential of the ∆rrn8 ε∆C mutant cells was significantly reduced, which led to a decrease in the cellular level of 70S ribosomes. The growth stalling was suppressed by adding glucose into the culture medium. Our findings suggest that the C-terminal region of the ε subunit is important for alleviating the temporal reduction in the membrane potential, by enhancing the ATP-dependent H+ -pumping activity of Fo F1 .


Cationic factors affecting phospholipase activities from human lung.
1. The effects of varying H+ and other cation concentrations on phospholipase activity were investigated on two particulate fractions from human lung, corresponding to the mitochondrial and microsomal fractions. 2. Three 14C-labelled substrates, arachidonyl-phosphatidylcholine (PC), -phosphatidylethanolamine (PE) and -phosphatidylinositol (PI) were used. 3. For two substrates, PE and PI, hydrolysis was maximal at pH 6, with either subcellular fraction. 4. Hydrolysis of all three substrates was strongly inhibited by EDTA and EGTA (10-25 mM). Addition of 2,2-dipyridyl, o-phenanthroline, 8-hydroxyquinoline or desferrioxamine (10 microM-1 mM) did not inhibit but often increased hydrolysis of all substrates. 5. Addition of Zn2+, as ZnCl2, (10 microM-1 mM) inhibited PE and PI, but not PC, hydrolysis. 6. The phospholipase activities from human lung appear to be dependent on Ca2+ for maximal activity and to be inhibited by other metal ions including Zn2+.


MR imaging strategies in primary and metastatic malignancy.
This article gives the rationale of the indications for the use of MRI in the evaluation of primary and metastatic malignancies of the neck, chest, abdomen, male and female pelvis, and musculoskeletal system. The imaging protocols for each of these areas is also included.


Efficacy of olanzapine in the treatment of psychosis in dementia with lewy bodies.
This study sought to determine whether patients with dementia with Lewy bodies (DLB) and psychosis responded to treatment with olanzapine. This was a post hoc analysis of a subgroup of patients with DLB included in a larger double-blind, placebo-controlled, randomized parallel group trial of olanzapine for the treatment of psychosis in patients with Alzheimer's disease. Patients meeting the consensus criteria for DLB and exhibiting parkinsonism and visual hallucinations were selected from the initial study. Psychosis was assessed with the Neuropsychiatric Inventory/Nursing Home (NPI-NH) version and the Brief Psychiatric Rating Scale (BPRS). Extrapyramidal symptoms were evaluated with the Simpson-Angus scale. Twenty-nine patients met the criteria for DLB; 10 were randomized to placebo, 5 received 5 mg of olanzapine, 7 received 10 mg of olanzapine and 7 received 15 mg of olanzapine. Patients with DLB treated with 5 mg of olanzapine showed significant reductions in delusions and hallucinations. Patients treated with 10 mg showed a significant reduction in the NPI-NH delusion subscale score. No significant differences were found between the 15-mg group and the placebo group. Confirmatory findings emerged from an analysis of the BPRS. Caregivers reported decreased disruptions in their occupational routines for the group receiving 5 or 10 mg of olanzapine. There was no significant exacerbation of parkinsonian symptoms in any study group, no decrement in Mini-Mental State Examination scores in any of the treatment groups, and symptoms suggestive of anticholinergic toxicity did not differ among treatment groups. This preliminary analysis suggests that olanzapine (5 or 10 mg) reduces psychosis in patients with DLB without worsening parkinsonism.


Laparoscopic resection of a retroperitoneal liposarcoma: a case report and review of the literature.
Here, we describe a case of a retroperitoneal liposarcoma successfully managed by laparoscopic surgery. A 72-year-old man underwent abnormal hypertension screening using computed tomography (CT), which revealed a low-density mass measuring 7 cm in diameter in the retroperitoneal space. We diagnosed the mass as a liposarcoma before the operation. The mass was resected laparoscopically. No perioperative complications were encountered, and the patient was discharged on the fourth postoperative day. The pathologic diagnosis was well-differentiated liposarcoma, indicating complete surgical resection. Thus, we conclude that a laparoscopic approach for the patients with retroperitoneal tumors, including liposarcomas, is safe, feasible, and minimally invasive, even if there is a possibility of the tumor being malignant.


Dural arteriovenous fistula at the foramen magnum: Report of a case and clinical-anatomical review.
Arterial supply and venous drainage at the foramen magnum is variable. Two main forms of clinical presentation, intracranial and spinal, can be differentiated when a dural arteriovenous fistula (DAVF) is found at this level. We describe a case of a 68-year-old patient with a progressive paraparesis, diagnosed of dural arteriovenous fistula located at the posterior lip of foramen magnum. We review, in this setting, the vascular radiological anatomy of those fistulas and its important correlation with neurologic clinical symptoms.


A Comparison of Pressure Mapping Between Two Pressure-Reducing Methods for the Sacral Region.
The objective of this study was to quantify tissue interface pressures that occurred in the sacrum, shoulder, and buttock/thigh regions while using (1) pillows or (2) a wedge system for off-loading of sacral pressures. Twenty-one volunteers (11 females and 10 males) residing near a Midwestern, university community consented to participate in the study. Testing was conducted in a hospital bed with pillows and a commercially available wedge system. Pressures were measured under 3 test conditions: (1) bed alone, (2) pillows positioned above and below the sacral region, and (3) wedges positioned above and below the sacral region. Each condition was tested with the elevation of the head of the bed at 3 different angles with respect to the foot of the bed (0°, 20°, and 30°). Two pressure mats were used to capture data; one was located in the shoulder region and the other in the sacral/buttocks/thigh region. Between the pillows and the wedge, the wedge system was the most effective in reducing pressures on the sacral area. In comparison to the bed condition, both the pillow and wedge conditions produced significantly lower mean sacral pressures than the bed alone (P < .05). Because the pillow and wedge systems tilted the body onto one side, higher pressures were identified on the buttock/thigh when compared to the bed alone (P < .05). Pressure reduction occurred in the sacral region with the pillow and wedge systems; however, this reduction resulted in increased pressures in the posterior-lateral regions of the buttocks and thighs. When using off-loading devices, consideration should be given to all factors, including tissue interface pressures on the sacrum, increased pressures on other body locations, and the likelihood that these increased pressures will result in tissue damage.


Otoneurological abnormalities in asymptomatic HIV-seropositive patients.
Twenty-four male patients, all homosexual except one, with asymptomatic HIV-infection were studied. The patients had no signs of opportunistic CNS-infections but 6 had been treated for syphilis. The patients were tested with auditory brainstem response (ABR) audiometry and with oculomotor tests (saccades and smooth pursuit eye movements). The ABR-recordings were pathological in 38% of the cases and the oculomotor tests in 50% of the cases. Abnormality of either one or of both methods were seen in 67% of the patients tested. The duration of the HIV-infection had no influence on the test results. The abnormal otoneurological tests indicate that occult CNS-dysfunction is a frequent finding in asymptomatic HIV-positive patients.


The oxidation-reduction and electrocatalytic properties of CO dehydrogenase from Oligotropha carboxidovorans.
CO dehydrogenase (CODH) from the Gram-negative bacterium Oligotropha carboxidovorans is a complex metalloenzyme from the xanthine oxidase family of molybdenum-containing enzymes, bearing a unique binuclear Mo-S-Cu active site in addition to two [2Fe-2S] clusters (FeSI and FeSII) and one equivalent of FAD. CODH catalyzes the oxidation of CO to CO2 with the concomitant introduction of reducing equivalents into the quinone pool, thus enabling the organism to utilize CO as sole source of both carbon and energy. Using a variety of EPR monitored redox titrations and spectroelectrochemistry, we report the redox potentials of CO dehydrogenase at pH 7.2 namely MoVI/V, MoV/IV, FeSI2+/+, FeSII2+/+, FAD/FADH and FADH/FADH-. These potentials are systematically higher than the corresponding potentials seen for other members of the xanthine oxidase family of Mo enzymes, and are in line with CODH utilising the higher potential quinone pool as an electron acceptor instead of pyridine nucleotides. CODH is also active when immobilised on a modified Au working electrode as demonstrated by cyclic voltammetry in the presence of CO.


Stoller afferent nerve stimulation in woman with therapy resistant over active bladder; a 1-year follow up.
In this prospective observational study, we investigated the efficacy of Stoller afferent nerve stimulation (SANS) in subjects with overactive bladder who failed anticholinergic treatment. Thirty-five subjects with overactive bladder who failed therapy with oxybutynin participated in this study. Treatment (n = 35) was given once a week for 30 minutes for overall 10 weeks. In treatment, SANS device (Urosurge) was used. Subjects were assessed with 3-day voiding diary, SEAPI quality of life questionnaires and cystometry before therapy after completion of therapy and at one-year follow-up. In 54% (n = 19) of subjects complete recovery was obtained after treatment. Urgency and SEAPI were reduced significantly whereas urine volume increased significantly (p < 0.01). Complete recovery was maintained in eight of the 19 subjects at one year. SANS treatment has a short-term positive effect in patients with resistant overactive bladder. However, it was also established that efficacy was maintained at 1 year in only 23% of subjects.


What is a genetic cause? The example of Alzheimer's Disease.
This paper focuses on the causation of diseases, particularly on the idea of a "genetic cause" taking Alzheimer's Disease (AD) as an example. We (1) provide some historical information and a synopsis of the current knowledge on the etiology and pathogenesis of AD, (2) analyse some conceptual problems related to the notion of "genetic disease", (3) elaborate on the alleged (genetic) cause of AD, and (4) place the discussion on the cause of AD in a broader philosophical context, paying attention to a constructivist perspective, the notions of causal connection and causal selection, and to some practical and normative consequences of our analysis. We conclude (a) that AD is not a specific disease entity with one specific cause, (b) that the idea of a single (sufficient) cause can still function as a heuristic tool in AD research and practice, and (c) that a "belief" in causation can go together with the notions of multicausality and probability.


Characterization of a novel Cry9Bb delta-endotoxin from Bacillus thuringiensis.
The Brazilian Bacillus thuringiensis serovar japonensis strain S725 was selected for its toxicity to the velvetbean caterpillar, Anticarsia gemmatalis. This strain produces spherical crystals harbouring a major protein of about 130 kDa which yields fragments of between 50 and 70 kDa upon trypsin activation. The protein showed a high level of identity and immunoafinity to the Cry9 class of delta-endotoxins. The cloned cry9-like gene sequence contains a 3492 bp ORF, which encodes a polypeptide of 1163 amino acids, with a predicted molecular mass of 131.4 kDa. The deduced amino acid sequence is unique and shows 73% identity to Cry9Ba, 64% identity to Cry9Ea, 63% identity to Cry9Da, and 59% identity to Cry9Ca proteins. The novel delta-endotoxin was assigned to a new subclass, Cry9Bb, by the Bt Toxin Nomenclature Committee. The Cry9Bb protein was expressed in an acrystalliferous Bt strain, and exhibited activity against the tobacco hornworm, Manduca sexta, and the velvetbean caterpillar, A. gemmatalis. The biological effect of an amino acid residue change, A84P, was investigated. The LC(50) for the Cry9Bb crystals against M. sexta neonate larvae was 6.84 microg/cm(2), while the LC(50) for the mutant's Cry9Bb crystals was 0.78 microg/cm(2). PCR screening revealed that in addition to cry9Bb, Bt strain S725 also contains cry1I and vip3 genes. Transcription analysis, using RT-PCR, showed that the cry1I gene was transcribed at T(2) and T(5) stages of sporulation.


[Systemic candidiasis in intensive care patients after antibiotic therapy].
The authors report fluconazole treatment of 19 I.C.U. patients in critical situations. 9 of these patients had septic multiorgan failure. All patients had received long term antibiotic treatment for at least seven days. The difficulties in clinical and laboratory diagnosis of systemic candidosis which requires treatment are discussed. All patients showed good response to fluconazole treatment concerning the opportunistic mycotic infection; none of the patients died because of systemic candidosis. Severe side effects were not observed.


Therapeutic transcatheter arterial embolization in the management of intractable haemorrhage from pelvic urological malignancies: preliminary experience and long-term follow-up.
To evaluate the feasibility, efficacy and complications of internal iliac artery embolization as a palliative measure in the control of intractable haemorrhage from advanced pelvic urological malignancies. Six patients (mean age 80 years, range 76-87) with advanced pelvic malignancies (three each with bladder carcinoma and prostate carcinoma) underwent embolization between September 1997 and July 2001, using permanent coils in the anterior division of internal iliac artery bilaterally. All patients had undergone conservative treatment before embolization for a mean of 2 days. The mean requirement for transfusion before embolization was 3.2 units. All patients except one were successfully embolized in a single setting with no complications. Bleeding was successfully controlled in one patient at the second attempt of embolization. There were minor complications, e.g. nausea, vomiting or fever, for a mean of 2 days, responding well to conservative treatment. At a mean follow-up of 22 months, no patient had a recurrence of bleeding. Internal iliac artery embolization is a feasible, effective and minimally invasive option in managing advanced pelvic urological malignancies presenting with intractable bleeding; it should be bilateral and permanent.


PCR-enzyme-linked immunosorbent assay for detection and identification of Campylobacter species: application to isolates and stool samples.
We report a PCR-enzyme-linked immunosorbent assay which identifies Campylobacter species by capture hybridization of a single-stranded 16S rRNA gene amplicon with species-specific probes in a microtiter plate format. Specificities were confirmed for both reference and field strains, but the type strain of Campylobacter coli was atypical. The assay was rapid, informative, and usable with stool-extracted DNA.


Impact of High-Power Short-Duration Radiofrequency Ablation on Long-Term Lesion Durability for Atrial Fibrillation Ablation.
The goal of this study was to compare lesion durability between high-power short-duration (HP-SD) and moderate-power moderate-duration (MP-MD) ablation strategies. HP-SD radiofrequency ablation (RFA) was developed to improve pulmonary vein isolation (PVI) by reducing the effect of catheter instability inherent to MP-MD ablation strategies. However, its long-term effect on lesion durability for the treatment of atrial fibrillation is unknown. Patients with atrial fibrillation (n = 112) underwent PVI using HP-SD ablation (45 to 50 W, 8 to 15 s) with contact force-sensing open irrigated catheter. Cavotricuspid isthmus, mitral annular, and roof lines were permitted. A control group (n = 112) underwent ablation using MP-MD ablation (20 to 40 W, 20 to 30 s) with similar technology. Chronic PV reconnection was examined in patients who required a redo procedure (HP-SD ablation, n = 18; MP-MD ablation, n = 23). The rate of PVI at the completion of the initial encirclement was similar between the HP-SD and MP-MD ablation strategies (90.2% vs. 83.0%; p = 0.006). The HP-SD strategy required shorter RFA time (17.2 ± 3.4 min vs. 31.1 ± 5.6 min; p < 0.001). The incidence of chronic PV reconnection was lower with HP-SD ablation (16.6% vs. 52.2%; p = 0.03). Areas of chronic reconnection were associated with catheter motion ≥1 mm for ≥50% application duration. In a higher proportion of HP-SD applications, catheter motion was <1 mm during ≥50% duration (88.6% vs. 72.8%; p < 0.001), allowing energy delivery with greater stability. Both ablation strategies were effective for cavotricuspid isthmus; however, the HP-SD strategy was less effective for mitral annular lines, requiring ablation at lower power for longer duration to avoid steam pops. HP-SD ablation may improve PVI durability, and it shortens RFA time. However, ablation in thicker myocardium often requires lower power applied for longer duration, allowing deeper lesions without tissue overheating.


The insect central complex.
Hordes of tourists flock to Washington, D.C. every spring to see the cherry trees blossom. Once in the city, they must find their way to the Tidal Basin where the Japanese trees grow. Fortunately, a number of visual landmarks can help them to navigate. In 1910, the United States Congress passed The Height of Buildings Act, limiting the elevation of commercial and residential structures in D.C. to 130 feet. Thus, the 555-foot-tall Washington Monument often looms large against the horizon, serving as an anchor point to help set the tourists' sense of direction. Once their heading is set, they can lose sight of the monument behind buildings or groups of tall Scandinavian visitors and still use their internal compass to navigate to the Basin. This compass keeps track of their paces and turns and updates their sense of where they are and where they need to go. Yet while their heading informs their actions, it does not dictate them. Tourists who have been to D.C. in the past can, for example, use remembered views to alter their routes to avoid crowds. On an even finer scale, their leg movements also depend on their current state - they might increase the frequency and length of their strides if hunger pangs compete with their desire to see cherry blossoms, for example. The way in which these disparate cues and motivations influence exploration is a neuroscience mystery across creatures large and small.


ACR Appropriateness Criteria® Clinically Suspected Vascular Malformation of the Extremities.
Vascular malformations of the extremities represent a wide spectrum of lesions, broadly divided into high-flow and low-flow categories. High-flow lesions include arteriovenous malformations and arteriovenous fistulas, while the more common low-flow lesions consist of venous and lymphatic malformations. The clinical presentation of vascular malformations is variable and can include extremity pain, discoloration, focal mass, or diffuse extremity enlargement. A vascular murmur can also be present and is more typical of high-flow lesions. While vascular malformations can often be diagnosed or strongly suspected by clinical features alone, imaging is often used to confirm the diagnosis, determine lesion characteristics and extent, and/or plan for treatment. Among the imaging options available, those usually appropriate for initial imaging of suspected vascular malformation are MR angiography without and with intravenous contrast, MRI without and with intravenous contrast, CT angiography with intravenous contrast, or US duplex Doppler. The American College of Radiology Appropriateness Criteria are evidence-based guidelines for specific clinical conditions that are reviewed annually by a multidisciplinary expert panel. The guideline development and revision include an extensive analysis of current medical literature from peer reviewed journals and the application of well-established methodologies (RAND/UCLA Appropriateness Method and Grading of Recommendations Assessment, Development, and Evaluation or GRADE) to rate the appropriateness of imaging and treatment procedures for specific clinical scenarios. In those instances where evidence is lacking or equivocal, expert opinion may supplement the available evidence to recommend imaging or treatment.


Mandibular fracture scoring system: for prediction of complications.
Mandibular fractures are one of the most commonly encountered injuries in trauma clinics. Although several widely accepted classification systems exist, these are mostly region specific, differ in the classification criteria used, and are sometimes only correlated with specific treatment modalities, thereby making it impossible to uniformly and comprehensively document facial fracture patterns. In this study, we developed a modified scoring system for mandibular fractures and analyzed the relationship between scoring of fractures that were treated and the incidence of complications after surgical treatment. To evaluate the suitability of the proposed scoring system, a prospective study on a series of 116 patients was performed. All the fractures were classified using the proposed scoring system. The scoring was based on clinical and radiological evaluation of each fracture. Patients were followed up postoperatively for presence of complications. A good correlation between the proposed scoring system and the incidence of complications was detected. This scoring system for mandibular fractures facilitates an objective and standardized assessment of the degree of severity of a fracture, thereby allowing for systematic evaluation of facial fracture outcomes, including assessment of complications. However, it is our understanding that a multicenter study should be performed before the effectiveness of the proposed classification can be clearly stated.


Investigations into seed dormancy in Grevillea linearifolia, G. buxifolia and G. sericea: anatomy and histochemistry of the seed coat.
Seeds of east Australian Grevillea species generally recruit post-fire; previous work showed that the seed coat was the controller of dormancy in Grevillea linearifolia. Former studies on seed development in Grevillea have concentrated on embryology, with little information that would allow testing of hypotheses about the breaking of dormancy by fire-related cues. Our aim was to investigate structural and chemical characteristics of the seed coat that may be related to dormancy for three Grevillea species. Seeds of Grevillea linearifolia, Grevillea buxifolia and Grevillea sericea were investigated using gross dissection, thin sectioning and histochemical staining. Water movement across the seed coat was tested for by determining the water content of embryos from imbibed and dry seeds of G. sericea. Penetration of intact seeds by Lucifer Yellow was used to test for internal barriers to diffusion of high-molecular-weight compounds. Two integuments were present in the seed coat: an outer testa, with exo-, meso- and endotestal (palisade) layers, and an inner tegmen of unlignified sclerenchyma. A hypostase at the chalazal end was a region of structural difference in the seed coat, and differed slightly among the three species. An internal cuticle was found on each side of the sclerenchyma layer. The embryos of imbibed seeds had a water content six times that of dry seeds. Barriers to diffusion of Lucifer Yellow existed at the exotestal and the endotestal/hypostase layers. Several potential mechanisms of seed coat dormancy were identified. The embryo appeared to be completely surrounded by outer and inner barriers to diffusion of high-molecular-weight compounds. Phenolic compounds present in the exotesta could interfere with gas exchange. The sclerenchyma layer, together with strengthening in the endotestal and exotestal cells, could act as a mechanical constraint.


Effective gene therapy for medullary thyroid carcinoma using recombinant adenovirus inducing tumor-specific expression of interleukin-12.
No satisfactory treatment of metastatic medullary thyroid carcinoma (MTC) is available. Cell-specific gene therapy offers a new approach. We have constructed a recombinant replication-defective adenoviral vector expressing murine interleukin-12 (mIL-12), driven by a modified CALC-I promoter (TCP). This vector (AdTCPmIL-12) includes two separate cassettes encoding mIL-12 p35 or p40 subunit controlled by TCP inserted in the E1 region of adenovirus type 5. In vitro and in vivo reporter gene expression using TCP revealed its cell-specific activity. AdTCPmIL-12-infected rat MTC (rMTC) cells produced high amounts of functional mIL-12 cells in vitro, while other cell lines infected with AdTCPmIL-12 did not. AdTCPmIL-12-transduced rMTC cells completely lost their tumorigenicity in syngenic WAG/Rij rats. Direct injection of 1 x 10(9) plaque forming units of AdTCPmIL-12 into subcutaneous rMTC tumors in WAG/Rij rats caused tumor regression in over 60% of animals within 20 days. Rats cured of tumors did not develop tumors after re-injection of naive rMTC cells, demonstrating lasting immunity. Treatment with AdTCPmIL-12 of one tumor resulted in regression of an established tumor at a distant site. Moreover, intratumoral or intravenous injection of AdTCPmIL-12 did not induce evident toxicity. These results indicate AdTCPmIL-12 can contribute to effective and less toxic gene therapy of MTC.


Identification of the novel HLA-DRB1*11:192 allele by sequence-based typing in Greece.
The new allele DRB1*11:192 exon 2 differs from the DRB1*11:01:01:01 by three substitutions.


Postoperative antibiotics are not associated with decreased wound complications among patients undergoing appendectomy for complicated appendicitis.
The objective of this study was to determine the role of postoperative antibiotics in reducing complications in patients undergoing appendectomy for complicated appendicitis. We performed a 5-year retrospective cohort study of adult patients who underwent appendectomy for acute appendicitis. Patients with complicated appendicitis (perforated or gangrenous) were analyzed on the basis of whether they received postoperative antibiotics. Main outcome measures were wound complications, length of stay (LOS), and readmission to hospital. Of 410 patients with complicated appendicitis, postoperative antibiotics were administered to 274 patients (66.8%). On univariate and multivariate analyses, postoperative antibiotics were not associated with decreased wound complications or readmission, but independently predicted an increased LOS (P = .01). Among patients with complicated appendicitis, postoperative antibiotics were not associated with a decrease in wound complications but did result in an increased hospital LOS.


Low-dose irinotecan improves advanced lupus nephritis in mice potentially by changing DNA relaxation and anti-double-stranded DNA binding.
Despite clear advances in the treatment of systemic lupus erythematosus (SLE), many patients still present with refractory lupus nephritis, requiring new treatment strategies for this disease. This study was undertaken to determine whether reduced doses of the topoisomerase I (topo I) inhibitor irinotecan, which is known as a chemotherapeutic agent, suppress SLE in (NZB × NZW)F1 (NZB/NZW) mice, and to evaluate the potential mechanism by which irinotecan influences the course of SLE. NZB/NZW mice were treated with low-dose irinotecan beginning at either 24 weeks of age or established glomerulonephritis, defined as proteinuria of grade ≥3+. Binding of anti-double-stranded DNA (anti-dsDNA) antibodies was measured by enzyme-linked immunosorbent assay (ELISA), and DNA relaxation was visualized by gel electrophoresis. Significantly reduced irinotecan doses improved lupus nephritis and prolonged survival in NZB/NZW mice. The lowest dose successfully used for the treatment of established murine lupus nephritis was >50 times lower than the dose usually used for chemotherapy in humans. As a mechanism, low-dose irinotecan reduced B cell activity. However, the levels of B cell activity in irinotecan-treated mice were similar to those in BALB/c mice of the same age, suggesting that irinotecan did not induce clear immunosuppression. In addition, incubation of dsDNA with topo I increased binding of murine and human anti-dsDNA antibodies, showing for the first time that relaxed DNA is more susceptible to anti-dsDNA antibody binding. This effect was reversed by addition of the topo I inhibitor camptothecin. Our findings indicate that topo I inhibition may be a novel and targeted therapy for SLE.


Dehydro-digitoxosides of digitoxigenin and digoxigenin: binding to beef heart (Na+ + K+)-ATPase in relation to unchanged digitoxosides.
Dehydro-digitoxosides are metabolites of digitalis glycosides. In order to study their possible biological activity their affinity to (Na+ + K+)-activated ATPase was determined and compared with unchanged glycosides. Based on the dissociation constants of glycoside-enzyme-complexes, the affinity of the dehydro-digitoxosides ranged in the same order of magnitude as that of the native glycosides. Comparing mono-, bis-, and tris-digitoxosides of digitoxigenin (dt-1, dt-2, dt-3) and of digoxin (dg-1, dg-2, dg-3) with the corresponding dehydrodigitoxosides (3'-dehydro-dt-1, 9'-dehydro-dt-2, 15'-dehydro-dt3, 3'-dehydro-dg-1 and 9'-dehydro-dg-2, respectively) the dehydro-digitoxosides had lower affinities to the enzyme. The highest dissociation constants (KD) were found for 3'-dehydro-dt-1 and 3'-dehydro-dg-1. The half maximal inhibition of (Na+ + K+)-ATPase activity (I50) coresponded to affinity measurements in all but two cases: dehydro-dt-3 and dehydro-dt-2 showed very low I50 values.


Arterial distensibility in patients with ruptured and unruptured intracranial aneurysms: is it a predisposing factor for rupture risk?
A risk factor assessment that reliably predicts whether patients are predisposed to intracranial aneurysm (IA) rupture has yet to be formulated. As such, the clinical management of unruptured IA remains unclear. Our aim was to determine whether impaired arterial distensibility and hypertrophic remodeling might be indicators of risk for IA rupture. The study population (n=49) was selected from consecutive admissions for either unruptured IA (n=23) or ruptured IA (n=26) from January to December 2010. Hemodynamic measures were taken from every patient, including systolic and diastolic blood pressure using a sphygmomanometer. Unruptured IA and ruptured IA characteristics, including aneurysmal shape, size, angle, aspect ratio, and bottleneck factor, were measured and calculated from transverse brain CT angiography images. With ultrasound, the right common carotid artery intima-media thickness was measured, as well as the lumen diameter during systole and diastole. Arterial wall strain, distensibility, stiffness index, and elastic modulus were calculated and compared between patients with unruptured IAs and ruptured IAs. A p-value less than 0.05 was considered statistically significant. General demographic data did not differ between patients with unruptured IAs and ruptured IAs. Greater mean intima-media thickness (p=0.013), mean stiffness index (p=0.044), and mean elastic modulus (p=0.026) were observed for patients with ruptured IAs. Moreover, mean strain (p=0.013) and mean distensibility (p=0.024) were decreased in patients with ruptured IAs. Patients with ruptured IAs demonstrated decreased arterial distensibility and increased intima-media thickness at the level of the carotid arteries. By measuring these parameters via ultrasound, it may be possible to predict whether patients with existing IAs might rupture and hemorrhage into the subarachnoid space.


Management of neuropsychiatric lupus.
Nervous system disease in systemic lupus erythematosus (SLE) is manifested by a wide variety of clinical manifestations. Despite the development of a universal classification for neuropsychiatric (NP) lupus in 1999, there continues to be considerable variability in the reported prevalence of NP syndromes between different lupus cohorts. Due to the lack of specificity of individual NP manifestations, non-SLE causes such as complications of therapy and co-morbidities must be considered in advance of attributing the event to one or more primary immunopathogenic mechanisms. These include intracranial microangiopathy, autoantibodies to neuronal and non-neuronal antigens, and the generation of proinflammatory cytokines and mediators. The diagnosis of NP-SLE remains largely one of exclusion and is approached in individual patients by thorough clinical evaluation, supported when necessary by autoantibody profiles, diagnostic imaging, electrophysiologic studies and objective assessment of cognitive performance. Given the diversity in clinical manifestations, the management is tailored to the specific needs of individual patients. In the absence of controlled studies, the use of symptomatic therapies, immunosuppressives, anticoagulants and non-pharmacologic interventions is supported by case series and clinical experience.


Inhibition of bile acid conjugation by cyclosporin A.
Each of the two steps involved in bile acid conjugation was tested in vitro for its sensitivity to inhibition by cyclosporin A (CsA). Bile acid-CoA: glycine/taurine N-acyltransferase, the enzyme which catalyzes the second step, was tested and found to be insensitive to inhibition by 20 microM CsA. Bile acid:CoA ligase, the enzyme which catalyzes the first step, was found to be inhibited by 25% at 10 microM CsA in the standard assay. The inhibition was competitive vs. bile acid and noncompetitive vs. ATP, and uncompetitive vs. CoA. CsA was also found to interfere with the divalent cation requirement of the enzyme at low concentrations of Mg2+ the maximum inhibition was 70%. The maximum inhibition obtainable at physiologic Mg2+ concentration was 40%. The extent of inhibition was presumably limited by the insolubility of CsA. At concentrations of CsA reached in vivo during drug therapy, CsA can be expected to significantly inhibit bile acid conjugation.


Reinduction of hormone sensitivity to goserelin following chemotherapy with vinorelbine in castration-resistant prostate cancer.
Primary androgen ablation leads to symptomatic improvement and a reduction in prostate-specific antigen (PSA) serum levels in patients with advanced prostate cancer, but all patients eventually become refractory to hormone therapy with progression of the disease and a life expectancy of about a year. We describe a patient who developed castration resistance, was treated with vinorelbine, and continues to be progression free on therapy with luteinizing hormone releasing hormone agonists alone, more than 2.5 years following cessation of treatment with vinorelbine.


Uridine alleviates carbon tetrachloride-induced liver fibrosis by regulating the activity of liver-related cells.
At present, liver fibrosis is a major challenge of global health. When hepatocyte regeneration cannot compensate for hepatocyte death, it will develop into liver fibrosis in chronic liver disease. Initially, collagen produced by myofibroblasts plays a role in maintaining liver integrity, but excessive collagen accumulation can inhibit the residual liver function, leading to liver failure. At present, many scientists are actively looking for drugs to alleviate liver fibrosis. In the current study, we investigated the potential role of uridine in the treatment of liver fibrosis (uridine is a plant/animal-derived pyrimidine nucleoside, therefore uridine can also be ingested and absorbed by the body, accompanied by the process of food intake). For this, we systematically studied the effect of uridine on CCl4-induced liver fibrosis in vitro and in vivo through a series of technologies, such as Western blot, laser confocal scanning microscope, ELISA and immunohistochemistry. The experimental results showed that uridine can effectively reduce the accumulation of collagen in liver. Furthermore, uridine can improve the activity of liver cells and alleviate CCl4-induced liver injury. Furthermore, uridine can significantly alleviate the risk factors caused by hepatic stellate cell activation, uridine treatment significantly down-regulated the expression of α-SMA, collagen type-I and fibronectin. In conclusion, the current research shows that uridine can alleviate CCl4-induced liver fibrosis, suggesting that uridine can be used as a potential drug to alleviate liver fibrosis.


Characterization and Genomic Analysis of Quinolone-Resistant Delftia sp. 670 Isolated from a Patient Who Died from Severe Pneumonia.
Antibiotic-resistant opportunistic pathogens have become a serious concern in recent decades, as they are increasingly responsible for hospital-acquired infections. Here, we describe quinolone-resistant Delftia sp. strain 670, isolated from the sputum of a patient who died from severe pulmonary infection. The draft genome sequence of this strain was obtained by whole-genome shotgun sequencing, and was subjected to comparative genome analysis. Genome analysis revealed that one critical mutation (Ser83Ile in gyrA) might play a decisive role in quinolone resistance. The genome of Delftia sp. strain 670 contains both type II and type VI secretion systems, which were predicted to contribute to the virulence of the strain. Phylogenetic analysis, assimilation tests, and comparative genome analysis indicated that strain 670 differed from the four known Delftia species, suggesting this strain could represent a novel species. Although the study could not determine the strain 670 as the pathogen led to mortality, our findings also presented the pathogenic potential of Delftia species, and the increasing severity of antibiotic resistance among emerging opportunistic pathogens. The whole genome sequencing and comparative analysis improved our understanding of genome evolution in the genus Delftia, and provides the foundation for further study on drug resistance and virulence of Delftia strains.


Bilio-pancreatic common channel in children. Clinical, biological and radiological findings in 12 children.
Twelve patients (11 girls and 1 boy) with dilated bile ducts and anomalous junction between the common bile duct and pancreatic duct are reported. All patients underwent preoperative opacification of the bile ducts either by transhepatic cholangiography or percutaneous cholecystography. Abdominal pain and jaundice were the main clinical symptoms. Reflux of pancreatic enzymes in the bile duct was proven by measuring amylase and lipase activity in the biliary system after IV injection of 1 IU/kg of cholecystokinin. All patients were operated upon. Bile ducts size returned to normal in all patients who are clinical well with a follow-up from 1 to 6 years.


Increased serum microRNA-29b expression and bad recovery in Japanese encephalitis virus infected patients; A new component to improve the disease recovery.
Japanese encephalitis virus (JEV) is a neurotropic mosquito-borne Flavivirus, mainly prevalent in Asia. It is the most important causative agent of acute viral encephalitis in humans. Recently, micro RNAs are discovered as a key regulator of inflammatory and immune responses in various diseases including neurological and viral infections. Thus, this study was proposed to check whether changes in cellular miRNA expression due to JE virus infection, can be detected in circulation which would be helpful in diagnosis and treatment. miRNAs (miR-29b and miR-146a) were analyzed in the serum of JEV infected patients using quantitative reverse transcription polymerase chain reaction (RT-qPCR). miR-146a was found significantly decreased (p = 0.0008) in JEV infected patients as compared to healthy controls whereas miR-29b was significantly increased (p = 0.001) in JEV patients recovered with neurological sequelae when compared to those recovered without sequelae. In conclusion, miRNA can be measured in serum. Studying microRNAs will provide novel information and help us to identify the components that can serve as biomarkers and can lead to new discovery in controlling disease recovery.


The beneficial effect of metformin on β-cell function in non-obese Chinese subjects with newly diagnosed type 2 diabetes.
Studies with metformin suggest a favourable change in β-cell function over sulphonylureas in the early course of obese type 2 diabetes mellitus (T2DM), but it remains unclear whether a similar effect is observed in non-obese individuals. Here we investigated the effects of metformin or glipizide gastrointestinal therapeutics system extended-release formulation (GITS) on β-cell function in non-obese patients with newly diagnosed T2DM. A total of 160 newly diagnosed patients with fasting glucose 7.0-13.0 mmol/L and body mass index <30 kg/m(2) from five centres in China were randomized to metformin or glipizide GITS for 24 weeks. Early insulin secretion [the ratio of area under the curve (AUC) of insulin to glucose during 0-30 min (InsAUC30 /GluAUC30 )] and insulin sensitivity [Matsuda index (ISIM )] were assessed during the standard meal tolerance test before and after therapy. Plasma glucagon-like peptide-1(GLP-1) and glucagon levels were also measured. Metformin improved InsAUC30 /GluAUC30 significantly (from 8.1 ± 0.6 pmol/mmol to 10.7 ± 0.7 pmol/mmol, p < 0.05), comparable to results with glipizide GITS. In the metformin-treated lean (body mass index < 25 kg/m(2) ) subgroup, the increase in ISIM was not significant, but the improvement in InsAUC30 /GluAUC30 was of great magnitude. Increased GLP-1 responses during meal tolerance test and decreased fasting glucagon level were observed after metformin treatment. Correlation analysis showed that the improvement of InsAUC30 /GluAUC30 was associated with the changes in HbA1c (r = -0.374, p = 0.000), ISIM (r = 0.356, p = 0.001), and ΔGLP-10-30 (r = 0.225, p = 0.02). Metformin improved β-cell function in non-obese subjects with newly diagnosed T2DM, which was partly independent of the change in insulin sensitivity in these subjects. This study provides evidence-based data to support metformin use in non-obese patients with T2DM as the first-line agent, which can improve both insulin sensitivity and β-cell function.


A systems approach identifies networks and genes linking sleep and stress: implications for neuropsychiatric disorders.
Sleep dysfunction and stress susceptibility are comorbid complex traits that often precede and predispose patients to a variety of neuropsychiatric diseases. Here, we demonstrate multilevel organizations of genetic landscape, candidate genes, and molecular networks associated with 328 stress and sleep traits in a chronically stressed population of 338 (C57BL/6J × A/J) F2 mice. We constructed striatal gene co-expression networks, revealing functionally and cell-type-specific gene co-regulations important for stress and sleep. Using a composite ranking system, we identified network modules most relevant for 15 independent phenotypic categories, highlighting a mitochondria/synaptic module that links sleep and stress. The key network regulators of this module are overrepresented with genes implicated in neuropsychiatric diseases. Our work suggests that the interplay among sleep, stress, and neuropathology emerges from genetic influences on gene expression and their collective organization through complex molecular networks, providing a framework for interrogating the mechanisms underlying sleep, stress susceptibility, and related neuropsychiatric disorders.


Measuring of time-series concentrations of nutrients in surface waters using osmotic sampler with air bubble segmentation and preservative addition.
The analysis of time-series concentrations (CTS) is of great importance when investigating the biogeochemical processes of nutrients in aquatic environments. However, obtaining CTS of nutrients remains a challenge using current sampling techniques. In this study, a novel in situ sampler was constructed using reverse osmosis membrane (ROM) osmotic pumps (OP) (ROM-OP sampler), and was used to obtain the CTS of nutrients in surface waters. The sampler consisted of a sampling OP, sample storing coil, filter, bubble injection module, and preservative adding module. When deployed, the sampling OP continuously draws ambient water through the filter into the sample storing coil, while simultaneously the preservative adding module continuously delivers preservative (H2SO4 solution) into the water flow. The bubble injection module periodically injects air bubbles into the sample storing coil, to segment the sample and create time stamp indicators that allow the sample age to be defined. Upon retrieval, the sample segments in the coil are sequentially pumped out of the sample storing coil and transferred into different vials for further analysis. The sampler was applied to measure the CTS of various nutrients, including dissolved total nitrogen, dissolved total phosphorus, dissolved reactive phosphorus, and nitrate in a river over a 20 day period and in municipal sewage treatment plant effluent for a 36 h period. Results showed that the ROM-OP sampler successfully obtained CTS of nutrients, capturing nutrient variations at a high temporal resolution. This sampler is relatively low-cost (~USD 300), small in size, lightweight, robust and does not require an external power source, showing high promise as an effective and efficient tool for monitoring nutrient CTS in aquatic environments.


Enhancement of optic nerve sheath on MRI in idiopathic intracranial hypertension(IIH).
Optic nerve sheath(ONS) enhancement in idiopathic intracranial hypertension (IIH) patients has been reported in recent years. In this retrospective observation, we analyzed the clinical characteristics of IIH patients with enhancement of ONS. Eighty-two patients with clinically diagnosed IIH from January 2017 to December 2019 were under observation. Then, based on the presence of contrast-enhancement (CE) in ONS on orbital magnetic resonance image (MRI), the IIH patients were divided into CE-ONS group and no-CE(NCE)-ONS group. Six months follow-up information was also included in the observation study. By comparing clinical data of the two groups of IIH patients, we tried to evaluate whether there is clinical heterogeneity in CE-ONS patients. 12 patients were included in CE-ONS group, 10 females and 2 males. 70 patients were included in NCE-ONS group, 56 women and 14 men. We found that patients with CE-ONS had a longer course of disease (median disease duration before diagnosis, 5 months vs. 3months, P<0.01) and more likely had the sign of distension of the perioptic subarachnoid space (DPSS) (58.33 % vs. 24.29 %, P = 0.034). But no significant differences were found in demographic characteristics, clinical symptoms, degree of visual impairment, papilledema, opening pressure(OP) on lumbar puncture and clinical outcomes. As a rare sign on MRI, ONS enhancement can occur in patients with IIH. IIH patients with CE-ONS may have a longer course of disease and more prone to DPSS, but there is no significant difference in clinical manifestations, OP, and clinical outcomes compared with IIH patients without CE-ONS.


Altered calcium metabolism in red blood cells of hypertensives: persistent marker or sequel of essential hypertension?
The characteristics of the increased calcium (Ca) influx observed in metabolically depleted red blood cells (RBCs) of hypertensive patients were investigated. Twenty-four normotensives, 16 untreated essential hypertensives, and 10 essential hypertensives under sufficient blood pressure control by 50-100 mg/day atenolol were studied. Free intracellular concentrations of Ca, sodium (Na), and potassium (K) were assessed using ion-selective electrodes in freeze-thawed RBCs, which were metabolically depleted by 30 mM desoxy-glucose at 37 degrees C for 48 h. In the treated hypertensives values for Ca and K at 24 and 48 h were not different from values for the normotensives, whereas elevated Ca was found in RBCs of untreated hypertensives. Na in treated hypertensives was significantly increased at 0 and 48 h, thus, being similar to values for untreated hypertensives. Additionally, RBCs of six normals were stressed in a glass/teflon potter. Before metabolic depletion electrolytes were not affected by this procedure, while Ca at 24 and 48 h of metabolic depletion increased to significantly higher values for the hypertensive patients as compared to the controls. These results suggest that the altered Ca metabolism in the RBCs of hypertensives may reflect a secondary phenomenon due to the mechanical damage to RBCs by the elevated blood pressure.


Altered mitochondrial respiration in selectively vulnerable brain subregions following transient forebrain ischemia in the rat.
Mitochondrial respiratory function, assessed from the rate of oxygen uptake by homogenates of rat brain subregions, was examined after 30 min of forebrain ischemia and at recirculation periods of up to 48 h. Ischemia-sensitive regions which develop extensive neuronal loss during the recirculation period (dorsal-lateral striatum, CA1 hippocampus) were compared with ischemia-resistant areas (paramedian neocortex, CA3 plus CA4 hippocampus). All areas showed reductions (to 53-69% of control) during ischemia for oxygen uptake rates determined in the presence of ADP or an uncoupling agent, which then recovered within 1 h of cerebral recirculation. In the ischemia-resistant regions, oxygen uptake rates remained similar to control values for at least 48 h of recirculation. After 3 h of recirculation, a significant decrease in respiratory activity (measured in the presence of ADP or uncoupling agent) was observed in the dorsal-lateral striatum which progressed to reductions of greater than 65% of the initial activity by 24 h. In the CA1 hippocampus, oxygen uptake rates were unchanged for 24 h, but were significantly reduced (by 30% in the presence of uncoupling agent) at 48 h. These alterations parallel the development of histological evidence of ischemic cell change determined previously and apparently precede the appearance of differential changes between sensitive and resistant regions in the content of high-energy phosphate compounds. These results suggest that alterations of mitochondrial activity are a relatively early change in the development of ischemic cell death and provide a sensitive biochemical marker for this process.


Anticarcinogenic effects of retinoids in animals.
Retinoids are effective inhibitors of chemical carcinogenesis in the skin, mammary gland, esophagus, respiratory tract, pancreas, and urinary bladder of experimental animals. Modification of the basic retinoid structure has produced retinoids with enhanced target organ specificity, resulting in increased anticancer activity with reduced systemic toxicity. Newer retinoidal benzoic acid derivatives are even more active. Combining retinoid treatment with other modulators of carcinogenesis results in a synergistic inhibition of tumor development. Retinoids in combination with hormonal manipulation are much more effective in inhibiting mammary carcinogenesis than is either treatment alone; this combination approach also inhibits mammary tumor recurrence following surgical removal of the first tumor. Retinoids are most effective when administered shortly after the carcinogenic insult. However, even when retinoid treatment is delayed, the compounds are still effective cancer chemopreventive agents for the mammary gland and urinary bladder. The time that retinoid exposure can be delayed and retain an anticancer effect is directly related to tumor latency, with a longer delay permissible against tumors with long latent periods. The mechanism(s) by which retinoids inhibit carcinogenesis is unknown; however, in the mammary gland, retinoids inhibit differentiation and proliferation, DNA synthesis, and RNA polymerase activity. Cytosolic retinoid-retinoid receptor complexing is apparently a prerequisite for the nuclear interaction of retinoids, at least in mammary cells.


Transfer of airway skills from manikin training to patient: success of ventilation with facemask or LMA-Supreme(TM) by medical students.
During emergency care, the ability to ventilate the patient's lungs is a crucial skill. Supraglottic airway devices have an established role in emergency care, and manikin trials have shown that placement is easy even for inexperienced users. However, there is current discussion as to what extent these results can be transferred to patients. We studied the transfer of skills learnt on a manikin to the clinical situation in novice medical students during their anaesthesia rotation. They were required to ventilate the lungs of a manikin using a facemask and then position a supraglottic airway device (LMA-Supreme™) and ventilate the lungs. This process was then repeated on anaesthetised patients, with standard ventilator settings to assess adequacy of ventilation. Sixty-three students participated in the manikin study. The success rate for ventilating the lungs was 100% for both devices, but the mean (SD) time to achieve successful ventilation was 27.8 (24.4) s with the facemask compared with 38.6 (22.0) s with the LMA-Supreme (p = 0.008). Fifty-one of the students progressed to the second part of the study. In anaesthetised patients, the success rate for ventilating the lungs was lower for the facemask, 27/41 (66%) compared with the LMA-Supreme 37/41 (90%, p = 0.006). For 26 students who succeeded with both devices, the tidal volume was lower using the facemask, 431 (192) ml compared with the LMA-Supreme 751 (221) ml (p = 0.001), but the time to successful ventilation did not differ, 60.0 (26.2) s vs 57.3 (26.6) s (p = 0.71). We conclude that the results obtained in manikin studies cannot be transferred directly to the clinical situation and that guidelines should take this into account. Based on our findings, a supraglottic airway device may be preferable to a facemask as the first choice for inexperienced emergency caregivers.


A generic tool for biotinylation of tagged proteins in transgenic mice.
The remarkable high affinity (Kd approximately 10(-15) M) of avidin/streptavidin for biotin has been extensively exploited in purification methodologies. Recently a small peptide sequence (Avi-tag) has been defined that can be specifically and efficiently biotinylated by the bacterial BirA biotin ligase. Fusion of this small peptide sequence to a protein of interest and co-expression with the BirA gene in mammalian cells allowed purification of the biotinylated protein together with its associated proteins and other molecules. Ideally, one would like to apply these technologies to purify tagged proteins directly from mouse tissues. To make this approach feasible for a large variety of proteins we developed a mouse strain that expresses the BirA gene ubiquitously by inserting it in the ROSA26 locus. We demonstrate that the BirA protein is indeed expressed in all tissues tested. In order to demonstrate functionality we show that it biotinylates the transgene-encoded Avi-tagged Gata1 and Oct6 transcription factors in erythroid cells of the foetal liver and Schwann cells of the peripheral nerve respectively. Therefore, this mouse can be crossed to any transgenic mouse to obtain efficient biotinylation of an Avi-tagged protein for the purpose of protein (complex) purification.


Is local participation always optimal for sustainable action? The costs of consensus-building in Local Agenda 21.
Is local participation always optimal for sustainable action? Here, Local Agenda 21 is a relevant case as it broadly calls for consensus-building among stakeholders. Consensus-building is, however, costly. We show that the costs of making local decisions are likely to rapidly exceed the benefits. Why? Because as the number of participants grows, the more likely it is that the group will include individuals who have an extreme position and are unwilling to make compromises. Thus, the net gain of self-organization should be compared with those of its alternatives, for example voting, market-solutions, or not making any choices at all. Even though the informational value of meetings may be helpful to policy makers, the model shows that it also decreases as the number of participants increase. Overall, the result is a thought provoking scenario for Local Agenda 21 as it highlights the risk of less sustainable action in the future.


Characterization of lactobacilli towards their use as probiotic adjuncts in poultry.
A total of 112 strains of lactic acid bacteria of duck origin were studied for their use as a probiotic feed supplement. In vitro studies included aggregation, co-aggregation, cell surface hydrophobicity and adhesion activities on poultry crop cells and human Hep2-cells. Additionally, growth with bile acids (chicken bile, ox gall and taurocholic acid) and tolerance to acidic pH were tested. Among all the isolates, two strains (Lactobacillus animalis TMW 1.972 and Lactobacillus salivarius TMW 1.992) were selected for a survival test in poultry. Monitoring and differentiation of these strains was achieved by selective detection as rifampicin and erythromycin double-resistant mutants. After a single feed administration, both micro-organisms were shown to persist in the crop and caecum of ducks for a period of 18 and 22 days, respectively. For identification of Lact. animalis and Lact. salivarius, two specific PCRs targeted against 16S rDNA were developed. Within the autochtoneous microflora of ducks, two strains of lactobacilli exhibited strong potential as probiotic adjuncts. The results indicate that the natural gut microflora of poultry serves as an excellent source for optimal strains. A general strategy for the selection of probiotic strains is presented. The suggested sequence of tests allows identification of the most promising candidates within complex ecosystems or large strain collections with minimal expenditure.


Reflection-mode TERS on Insulin Amyloid Fibrils with Top-Visual AFM Probes.
Tip-enhanced Raman spectroscopy provides chemical information while raster scanning samples with topographical detail. The coupling of atomic force microscopy and Raman spectroscopy in top illumination optical setup is a powerful configuration to resolve nanometer structures while collecting reflection mode backscattered signal. Here, we theoretically calculate the field enhancement generated by TER spectroscopy with top illumination geometry and we apply the technique to the characterization of insulin amyloid fibrils. We experimentally confirm that this technique is able to enhance the Raman signal of the polypeptide chain by a factor of 105, thus revealing details down to few molecules resolution.


Comparison of the distribution of binding sites for the potassium channel ligands [125I]apamin, [125I]charybdotoxin and [125I]iodoglyburide in the rat brain.
Potassium channels represent a diverse and promising target for drug development. Pharmacological subtypes of K channels have begun to emerge based on the development of both organic molecules and peptide toxins which possess subtype selectivity. In order to evaluate the neuroanatomical distribution of these subtypes we have utilized the ligands [125I]apamin, [125I]charybdotoxin and [125I]iodoglyburide in an autoradiographic study of rat brain. In the rat brain, these ligands have selectivity for the low conductance Ca(2+)-activated, voltage-gated K channels and ATP-sensitive K channels respectively. The distribution of binding sites for these three ligands were distinctly different. [125I]Apamin binding was highest in various thalamic and hippocampal structures, while only low to moderate levels of [125I]charybdotoxin binding were seen in these regions. In contrast, very high levels of [125I]charbydotoxin were seen in white matter regions such as the lateral olfactory tract and fasciculus retroflexus. High levels of [125I]charybdotoxin binding were also seen in gray matter-containing regions such as the zona incerta, medial geniculate and superior colliculus, where low to moderate [125I]apamin binding was found. [125I]Iodoglyburide presented a more uniform binding with the highest levels in the globus pallidus, islands of Calleja, anteroventral nucleus of the thalamus and zonas reticulata of the substantia nigra. These results indicate that subtypes of K channels have very different distributions in the brain. As such, the results imply differing CNS actions for potential modulators of K channel subtypes.