Problems of identification in the clinical laboratory, state of the art and other considerations.
In recent years, there have been no new commercial developments in identification methods for the clinical chemical laboratory. We are still faced, however, with the problem of ensuring accurate identification. By using a model, the importance of reliable identification can be shown mathematically. The complete identification of a sample is based on a five point identification statement. As a rule, appropriate measures are taken to abridge this identification process, depending on the local situation. The terminology of identification processes is defined and the presently available techniques are discussed.


The F19W mutation reduces the binding affinity of the transmembrane Aβ11-40 trimer to the membrane bilayer.
Alzheimer's disease is linked to the aggregation of the amyloid-β protein (Aβ) of 40 or 42 amino acids. Lipid membranes are known to modulate the rate and mechanisms of the Aβ aggregation. Point mutations in Aβ can alter these rates and mechanisms. In particular, experiments show that F19 mutations influence the aggregation rate, but maintain the fibril structures. Here, we used molecular dynamics simulations to examine the effect of the F19W mutation in the 3Aβ11-40 trimer immersed in DPPC lipid bilayers submerged in aqueous solution. Substituting Phe by its closest (non-polar) aromatic amino acid Trp has a dramatic reduction in binding affinity to the phospholipid membrane (measured with respect to the solvated protein) compared to the wild type: the binding free energy of the protein-DPPC lipid bilayer increases by 40-50 kcal mol-1 over the wild-type. This is accompanied by conformational changes and loss of salt bridges, as well as a more complex free energy surface, all indicative of a more flexible and less stable mutated trimer. These results suggest that the impact of mutations can be assessed, at least partially, by evaluating the interaction of the mutated peptides with the lipid membranes.


Constructed molecular sensor to enhance metal detection by bacterial ribosomal switch-ion channel protein interaction.
Molecular biosensors are useful tools that detect metal ions or other potentially toxic chemicals. However, the efficiency of conventional sensors is limited in mixed metals substrates, which is the common way they are found in nature. The use of biosensors constructed from genetically modified living microbial systems has the potential of providing sensitive detection systems for specific toxic targets. Consequently, our investigation was aimed at assembling different genetic building blocks to produce a focused microbial biosensor with the ability to detect specific metals. This objective was achieved by using a synthetic biology approach. Our genetic building blocks, including a synchronized ribosomal switch-iron ion channel, along with sequences of promoters, metal-binding proteins (Fe, Pb), ribosomal binding sites, yellow fluorescence reporter protein (YFRP), and terminators, were constructed within the same biobrick in Escherichia coli. We used an rpoS ribosomal switch containing an aptamer, which responds to the specific metal ligands, in synchronization with an iron ion channel, TonB. This switch significantly stimulates translation, as expressed by higher fluorescence, number of colonies, and concentration of RNA in E. coli. The positive results show the effectiveness of using genetically tailored synchronized ribosomal switch-ion channels to construct microbial biosensors to detect specific metals, as tested in iron solutions.


[DHEAS/cortisol index as a marker of stress-induced premature aging.]
The detection of stress-induced premature aging with the aim of correcting damaging adaptive effects seems to be actual today. The article discusses the use of the DHEAS / cortisol index as a screening marker for accelerating the rate of biological aging in people exposed to stress.


Use of Interaction Energies in QM/MM Free Energy Simulations.
The use of the most accurate (i.e., QM or QM/MM) levels of theory for free energy simulations (FES) is typically not possible. Primarily, this is because the computational cost associated with the extensive configurational sampling needed for converging FES is prohibitive. To ensure the feasibility of QM-based FES, the "indirect" approach is generally taken, necessitating a free energy calculation between the MM and QM/MM potential energy surfaces. Ideally, this step is performed with standard free energy perturbation (Zwanzig's equation) as it only requires simulations be carried out at the low level of theory; however, work from several groups over the past few years has conclusively shown that Zwanzig's equation is ill-suited to this task. As such, many approximations have arisen to mitigate difficulties with Zwanzig's equation. One particularly popular notion is that the convergence of Zwanzig's equation can be improved by using interaction energy differences instead of total energy differences. Although problematic numerical fluctuations (a major problem when using Zwanzig's equation) are indeed reduced, our results and analysis demonstrate that this "interaction energy approximation" (IEA) is theoretically incorrect, and the implicit approximation invoked is spurious at best. Herein, we demonstrate this via solvation free energy calculations using IEA from two different low levels of theory to the same target high level. Results from this proof-of-concept consistently yield the wrong results, deviating by ∼1.5 kcal/mol from the rigorously obtained value.


Effects of exogenous MSH on the transformation from phaeo- to eumelanogenesis within C57BL/6J-Ay/a hairbulb melanocytes.
The extent to which alpha melanocyte-stimulating hormone (MSH) is a true in vivo regulator of melanogenesis in mice is unknown. The objective of this study was to determine if MSH-induced eumelanogenesis in hairbulb melanocytes of yellow (Ay/a) mice mimics the natural program of eumelanogenesis occurring in genetically black (a/a) hairbulb melanocytes. We conducted quantitative transmission electron microscopy on melanosome differentiation within MSH-treated regenerating 9-d hairbulbs of Ay/a and a/a mice. Results of exogenous alpha-MSH injections (5 d at 0.15 mM MSH) showed that the striking visual darkening of hair was accompanied by an incomplete transformation of phaeo- to eumelanogenesis. Ontogenetic data on developmental stages I-IV of 3678 melanosomes based on geometric considerations (length, width, shape, and area) showed that MSH did not induce a complete transformation from spherical phaeomelanosomes to elliptical eumelanosomes. Also, observations on the number of vesiculoglobular bodies and matrix organization reveled that MSH-treated Ay/a melanosomes retained distinct features of phaeomelanogenesis even after 5 d of MSH treatment. Thus, MSH induced a partial but incomplete pattern of eumelanogenesis in regenerating hairbulb melanocytes of Ay/a mice. The continued investigation of the dynamics of melanin synthesis in MSH-induced Ay/a mice melanocytes possessing "mosaic" melanosomes could be productive in understanding fundamental relationships between tyrosinase activity, matrix function, matrix structure, and regulation of melanin (phaeo- and/or eumelanin) synthesis.


Readiness to recognize: A qualitative case study of educational identification of TBI.
The educational identification (ED-ID) of students with traumatic brain injury (TBI) has been legislated in numerous states, and TBI has become its own special education category. Questions abound as to whether these changes will lead to more readily identifying these students to provide appropriate special education supports. Elucidate one state's response to legislative changes around ED-ID of TBI. This is a phenomenological case study of nine school districts in one state. Researchers conducted focus groups as well as individual interviews with various special education staff. On-going analysis of the focus group and interview data occurred, until themes were finalized. Reactions of staff comprised two opposing viewpoints: those supporting and carrying out ED-ID ("Toward ED-ID") and those describing processes and attitudes that stand in the way ("Barriers"). Themes that were supportive of ED-ID include Credible History, Personnel, Targeted Intervention, Effective Communication, and Education. Themes aligned with the barrier viewpoint include Concussion Protocols, Medical Nature, Misidentification, Stigma, and Communication Breakdown. While many districts reported success with ED-ID for TBI, responses to the ED-ID legislation and TBI category were mixed, with enough barriers identified to cause concern over whether the legislative and policy changes have been overall beneficial.


Vibrationally Coherent Preparation of the Transition State for Photoisomerization of the Cyanine Dye Cy5 in Water.
Femtosecond pump-continuum probe spectroscopy with impulsive excitation was employed to observe coherent wavepacket motions of the cyanine dye Cy5 in water that promote photoisomerization after optical preparation of the first excited singlet state, S1. The chief component in the excited-state vibrational coherence is a resonance Raman-inactive, 273 cm(-1) mode of mixed carbon-carbon bond length alternation and out-of-plane or twisting character. The ultrafast (30 fs) damping of these motions arises from trajectories that irreversibly cross the transition state barrier; after several recurrences to the transition state region, vibrational cooling traps a significant fraction of the excited-state molecules in the planar, Franck-Condon region of the potential energy surface. Motion in the 273 cm(-1) promoting mode is apparently launched by a change in conformation of the conjugated polyene backbone during the first few vibrations of the high-frequency C-C and C═C bond length alternation coordinates that principally contribute to the initial displacement from the Franck-Condon structure. To our knowledge, this work provides the first direct observations of the intramolecular redistribution of excited-state potential energy into reactive motions that are rapidly damped by transition state barrier-crossing events leading to photoisomerization in a conjugated polyene. These results provide insight into the vibrational dynamics that contribute to the photoisomerization of retinal protonated Schiff bases in the rhodopsins and to the formation of intramolecular charge transfer character in carotenoids in photosynthetic light-harvesting proteins.


Multi-Component Analysis of Ilex Kudingcha C. J. Tseng by a Single Marker Quantification Method and Chemometric Discrimination of HPLC Fingerprints.
The quantitative analysis of multiple components with a single marker (QAMS) method was firstly established for simultaneous determination of 18 active components in Ilex kudingcha C. J. Tseng by HPLC. Using rutin, isochlorogenic acid A and kudinoside A as internal refererence substances (IRS), compatibility results showed that the relative correction factors (RCFs) of all compounds showed good reproducibility under different chromatographic conditions. On the basis of previous studies, the accuracy of the QAMS method was systematically evaluated by investigating the influences of curve intercept, analytes and IRS concentration. The results showed that the concentration (especially at low level) of analytes and curve intercept were the major influencing parameters for the LRG-QAMS method (LRG = linear regression), whereas the influence of IRS concentration seemed more apparent in terms of the AVG-QAMS method (AVG = average). The two approaches were complementary with each other. In addition, hierarchical clustering analysis (HCA), principal components analysis (PCA) and similarity analysis (SA) were performed to differentiate and classify the samples based on the contents of 18 marker compounds. The results of the different chemometric analyses were completely consistent with each other, and could be supported by the quantification results.


Culture and characterization of microvascular endothelial cells derived from human brain.
Primary cultures of human cerebral endothelial cells were established from microvessels isolated from cortical fragments removed at surgery for seizure disorder and from brains at autopsy. A uniform population of cells growing in close association to each other formed confluent monolayers by 7 to 10 days in culture. They contained factor VIII/Von Willebrand antigen, the most specific marker for cells of endothelial origin, and showed lectin-binding sites for Ulex europaeus agglutinin characteristic of human endothelium. Cultured cells formed thin, continuous monolayers, contained few pinocytotic vesicles, and were joined together by tight junctional complexes. More than 99% of the intercellular junctions restricted the transendothelial passage of horseradish peroxidase. Monolayers of human brain microvessel endothelial cells thus resemble cerebral endothelium in vivo and should provide a useful in vitro model for studies of the biology of these cells and their role in the pathogenesis of certain human central nervous system diseases associated with abnormal blood-brain barrier function.


Electrical impedance plethysmography technique in estimating pulmonary function status.
An attempt has been made to study the appropriateness of electrical impedance plethysmography (EIP) in detecting pulmonary health status. A feasibility study was conducted on normal, obstructive and restrictive subjects aged between 20 and 60 years. Quantitative assessment of pulmonary function was made by way of both EIP and pulmonary function tests (PFT). Amongst the various EIP parameters, a statistically significant difference was observed for the respiratory band power, between normal (129.7) and obstructive (35.8) subjects, indicating that this EIP variable can be used to distinguish between the two pulmonary function states. A significant positive correlation was observed between a spirometry parameter, peak expiratory flow rate (PEFR), and an EIP parameter, respiratory amplitude (r = 0.372, p < 0.05), thereby indicating that EIP information by way of respiratory amplitude is comparable to that provided by PEFR. Hence, both respiratory amplitude and respiratory band power were seen to provide useful information on pulmonary health status.


Surgical incision induces learning impairment in mice partially through inhibition of the brain-derived neurotrophic factor signaling pathway in the hippocampus and amygdala.
Surgical incision-induced nociception contributes to the occurrence of postoperative cognitive dysfunction. However, the exact mechanisms involved remain unclear. Brain-derived neurotrophic factor (BDNF) has been demonstrated to improve fear learning ability. In addition, BDNF expression is influenced by the peripheral nociceptive stimulation. Therefore, we hypothesized that surgical incision-induced nociception may cause learning impairment by inhibiting the BDNF/tropomyosin-related kinase B (TrkB) signaling pathway. The fear conditioning test, enzyme-linked immunosorbent assay, and Western blot analyses were used to confirm our hypothesis and determine the effect of a plantar incision on the fear learning and the BDNF/TrkB signaling pathway in the hippocampus and amygdala. The freezing times in the context test and the tone test were decreased after the plantar incision. A eutectic mixture of local anesthetics attenuated plantar incision-induced postoperative pain and fear learning impairment. ANA-12, a selective TrkB antagonist, abolished the improvement in fear learning and the activation of the BDNF signaling pathway induced by eutectic mixture of local anesthetics. Based on these results, surgical incision-induced postoperative pain, which was attenuated by postoperative analgesia, caused learning impairment in mice partially by inhibiting the BDNF signaling pathway. These findings provide insights into the mechanism underlying surgical incision-induced postoperative cognitive function impairment.


Incidence of Bradycardia and Outcomes of Patients Who Underwent Orbital Atherectomy Without a Temporary Pacemaker.
We analyzed the incidence of bradycardia and the safety of patients with severely calcified coronary lesions who underwent orbital atherectomy without the insertion of a temporary pacemaker. The presence of severely calcified coronary lesions can increase the complexity of percutaneous coronary intervention due to the difficulty in advancing and optimally expanding the stent. High-pressure inflations to predilate calcified lesions may cause angiographic complications like perforation and dissection. Suboptimal stent expansion is associated with stent thrombosis and restenosis. Orbital atherectomy safely and effectively modifies calcified plaque to facilitate optimal stent expansion. The incidence of bradycardia in orbital atherectomy is unknown. Fifty consecutive patients underwent orbital atherectomy from February 2014 to September 2016 at our institution, none of whom underwent insertion of a temporary pacemaker. The final analysis included 47 patients in this retrospective study as 3 patients were excluded because of permanent pacemaker implantation. The primary endpoint was significant bradycardia, defined as bradycardia requiring emergent pacemaker placement or a heart rate <50 bpm at the end of atherectomy. The primary endpoint occurred in 4% of all patients, all driven by patients who experienced a heart rate decreasing to <50 bpm. The major adverse cardiac and cerebral event rate was 6%, driven by death (2%) and myocardial infarction (4%). No patient experienced target-vessel revascularization, stroke, or stent thrombosis. Angiographic complications included perforation in 2%, slow-flow in 4%, and flow-limiting dissection in 0%. Significant bradycardia was uncommon during orbital atherectomy. Performing orbital atherectomy without a temporary pacemaker appears to be safe.


The detection, assessment and clinical evolution of interstitial lung abnormalities identified through lung cancer screening.
Interstitial lung abnormalities (ILAs) are common incidental findings in lung cancer screening; however, their clinical evolution and longer-term outcomes are less clear. The aim of this cohort study was to report 5-year outcomes of individuals with ILAs identified through a lung cancer screening programme. In addition, we compared patient-reported outcome measures (PROMs) in patients with screen-detected ILAs to newly diagnosed interstitial lung disease (ILD) to assess symptoms and health-related quality of life (HRQoL). Individuals with screen-detected ILAs were identified, and 5-year outcomes, including ILD diagnoses, progression-free survival and mortality, were recorded. Risk factors associated with ILD diagnosis were assessed using logistic regression and survival using Cox proportional hazard analysis. PROMs were compared between a subset of patients with ILAs and a group of ILD patients. 1384 individuals underwent baseline low-dose computed tomography screening, with 54 (3.9%) identified as having ILAs. 22 (40.7%) were subsequently diagnosed with ILD. 14 (25.9%) individuals died, and 28 (53.8%) suffered disease progression within 5 years. Fibrotic ILA was an independent risk factor for ILD diagnosis, mortality and reduced progression-free survival. Patients with ILAs had lower symptom burden and better HRQoL in comparison to the ILD group. Breathlessness visual analogue scale (VAS) score was associated with mortality on multivariate analysis. Fibrotic ILA was a significant risk factor for adverse outcomes including subsequent ILD diagnosis. While screen-detected ILA patients were less symptomatic, breathlessness VAS score was associated with adverse outcomes. These results could inform risk stratification in ILA.


Downregulation of female doublesex expression by oral-mediated RNA interference reduces number and fitness of Anopheles gambiae adult females.
Mosquito-borne diseases affect millions worldwide, with malaria alone killing over 400 thousand people per year and affecting hundreds of millions. To date, the best strategy to prevent the disease remains insecticide-based mosquito control. However, insecticide resistance as well as economic and social factors reduce the effectiveness of the current methodologies. Alternative control technologies are in development, including genetic control such as the sterile insect technique (SIT). The SIT is a pivotal tool in integrated agricultural pest management and could be used to improve malaria vector control. To apply the SIT and most other newer technologies against disease transmitting mosquitoes, it is essential that releases are composed of males with minimal female contamination. The removal of females is an essential requirement because released females can themselves contribute towards nuisance biting and disease transmission. Thus, females need to be eliminated from the cohorts prior to release. Manual separation of Anopheles gambiae pupae or adult mosquitoes based on morphology is time consuming, is not feasible on a large scale and has limited the implementation of the SIT technique. The doublesex (dsx) gene is one of the effector switches of sex determination in the process of sex differentiation in insects. Both males and females have specific splicing variants that are expressed across the different life stages. Using RNA interference (RNAi) to reduce expression of the female specific (dsxF) variant of this gene has proven to have detrimental effects to the females in other mosquito species, such as Aedes aegypti. We tested oral RNAi on dsx (AgdsxF) in An. gambiae. We studied the expression pattern of the dsx gene in the An. gambiae G3 strain. We knocked down AgdsxF expression in larvae through oral delivery of double stranded RNA (dsRNA) produced by bacteria and observed its effects in adults. Our results show that feeding of AgdsxF dsRNA can effectively reduce (> 66%) the mRNA of female dsx transcript and that there is a concomitant reduction in the number of female larvae that achieve adulthood. Control groups produced 52% (± 3.9% SE) of adult males and 48% (± 4.0% SE) females, while AgdsxF dsRNA treated groups had 72.1% (± 4.0% SE) males vs 27.8% females (± 3.3% SE). In addition, the female adults produce fewer progeny, 37.1% (± 8.2% SE) less than the controls. The knockdown was sex-specific and had no impact on total numbers of viable male adults, in the male dsx transcripts or male fitness parameters such as longevity or body size. These findings indicate that RNAi could be used to improve novel mosquito control strategies that require efficient sex separation and male-only release of An. gambiae by targeting sex determination genes such as AgdsxF. The advantages of using RNAi in a controlled setting for mosquito rearing are numerous, as the dose and time of exposure are controlled, and the possibility of off-target effects and the waste of female production would be significantly reduced.


The determinants of health-related quality of life among patients with newly diagnosed lung cancer in Taiwan: A cross-sectional study.
Although considered one of the most important prognostic factors for lung cancer patients, the health-related quality of life (HRQOL) of the newly diagnosed lung cancer population remains scarcely focused on in the literature. Therefore, we aimed to identify the determinants of HRQOL among newly diagnosed lung cancer patients in Taiwan. Two hundred and fifty patients newly diagnosed with lung cancer were recruited from a medical center in northern Taiwan through convenience sampling. Four structured questionnaires, including the Taiwanese version of the MD Anderson symptom inventory (MDASI-T), the Taiwanese version of the Pittsburgh Sleep Quality Index (PSQI-T), the International Physical Activity Questionnaire-Short Form (IPAQ-SF), and the World Health Organization Quality of Life-BREF (WHOQOL-BREF), were used to collect data. Further, a multivariate stepwise linear regression was conducted to determine the independent risk factors for HRQOL. A p value of less than 0.05 was considered statistically significant. The patients (mean age was 61.04 years, 51.2% male, 94.0% non-small-cell lung cancer, 56.4% stage IIIB-IV) had moderate levels of HRQOL among the physical, psychological, social, and environmental domains, as well as overall QOL. HRQOL was not correlated with married status, religion, and comorbidity. Gender, age, family income, smoking status, cancer stage, ECOG PS scores, PA, symptom burden (severity and interference), and PSQI global scores were correlated with HRQOL. Notably, symptom severity was the dominant negative predictor affecting the psychological and environmental domains of QOL (β = -4.313 and -3.500, respectively), accounting for 23.2% and 14.6% of the variance, respectively. On the other hand, symptom interference was the dominant negative predictor affecting the physical and social domains of QOL, as well as overall QOL (β = -3.592, -1.984, and -0.150, respectively), accounting for 44.4%, 15.0%, and 24.1% of the variance, respectively. Newly diagnosed lung cancer patients suffered symptom severity and interference that significantly impaired their HRQOL; particularly, symptom interference affected the physical domain of QOL. Healthcare professionals should pay more attention to cancer-related symptom severity, symptom interference, and HRQOL changes when caring for newly diagnosed lung cancer patients.


The six critical attributes of the next generation of quality management software systems.
Driven by both the need to meet regulatory requirements and a genuine desire to drive improved quality, quality management systems encompassing standard operating procedure, corrective and preventative actions and related processes have existed for many years, both in paper and electronic form. The impact of quality management systems on 'actual' quality, however, is often reported as far less than desired. A quality management software system that moves beyond formal forms-driven processes to include a true closed loop design, manage disparate processes across the enterprise, provide support for collaborative processes and deliver insight into the overall state of control has the potential to close the gap between simply accomplishing regulatory compliance and delivering measurable improvements in quality and efficiency.


Prevalence and incidence of bloodborne viral infections among Danish prisoners.
In order to determine the prevalence and incidence of bloodborne viral infections among prisoners, we conducted a prospective study in a Danish medium security prison for males. The prisoners were offered an interview and blood test for hepatitis and human immunodeficiency virus HIV at inclusion as well as at release from prison or end of study. Of 403 prisoners available 325 (79%) participated in the initial survey and for 142 (44%) a follow-up test was available. 43% (140/325) of the participants were injecting drug users (IDUs) of whom 64% were positive for hepatitis B (HBV) and 87% for hepatitis C (HCV) markers. No cases of HIV or human T lymphotropic virus (HTLV) were found. 32% of all prisoners could transmit HBV and/or HCV by blood contact. 70% of IDUs had shared injecting equipment, and 60% had injected inside prison. Only 2% of IDUs were vaccinated against HBV. Duration of injecting drug use, numbers of imprisonments, and injecting in prison were independently and positively associated with the presence of HBV antibodies among IDUs by logistic regression analysis. The HBV incidence was 16/100 PY (95% CI: 2-56/100 PY) and the HCV incidence 25/100 PY (1-140) among injecting drug users (IDUs). We conclude that IDUs in prison have an incidence of hepatitis B and C 100 times higher than reported in the general Danish population. They should be vaccinated against hepatitis B and new initiatives to stop sharing of injecting equipment in and outside prison is urgently needed.


Total synthesis of (±)-communesin F via a cycloaddition with indol-2-one.
A concise total synthesis of (±)-communesin F has been completed in 15 linear steps from 4-bromotryptophol in an overall yield of 6.7%. A key step features the cycloaddition of indol-2-one with 3-(2-azidoethyl)-4-bromoindole and facilitates the rapid construction of the lower aminal-containing tetracyclic core of the natural product.


Temporal and steric analysis of ionic permeation and binding in SERCA via molecular dynamic simulations.
The availability of the crystal structure of the sarco(endo)plasmic reticulum calcium ATPase (SERCA) has allowed atomic-level molecular dynamic (MD) simulations of this membrane transport protein to be done. The biomedical and nanotechnological implications of this work are discussed as well as the methods of performing the simulations and analysis. We have performed nanosecond timescale simulations of SERCA for several of its known conformations in a lipid/water environment. One simulation contained Ca(2+) ions, while others without ions were analyzed by techniques such as steric pathway determination. We discuss details of the resulting putative cytoplasmic and lumenal pathways, along with experimental evidence from the literature to support our conclusions. Finally, we give a brief overview of future research directions, as they pertain to MD simulations and their analysis. The methodology used in this work shows that significant insight into the structure-function relationship of ion-motive transmembrane pumps can be derived by a combination of simulation tools and analysis techniques including MD trajectories, steric analysis and electrostatic potentials.


A simplified modification of the AOAC official method for determination of total dietary fiber using newly developed enzymes: preliminary interlaboratory study.
A preliminary interlaboratory study was conducted to evaluate the validity of the modified AOAC method for determination of total dietary fiber by Tada and Innami, in which the 3-step enzymatic digestion process in AOAC Method 991.43 is modified to a 2-step process without pH adjustment. Total dietary fiber contents in 8 representative foodstuffs were measured using both the original AOAC Method 991.43 and the modified method in 6 research facilities in Japan. Repeatability relative standard deviations, reproducibility relative standard deviations, and Horwitz ratio values from the modified method were equivalent to those from AOAC Method 991.43, except in the rice sample. However, this exceptional case shown in the modified method was entirely dissolved by the addition of alpha-amylase stabilizing agents. The modified method, which shortens the process of enzymatic digestion from 3 to 2 steps and in which only reaction temperature is adjusted under the same pH, was found not only to give accurate values comparable to the original method, but also to substantially reduce the labor required by the laboratory staff in the process of routine analysis. This study revealed that the validity of the modified method was further ensured by adding alpha-amylase stabilizing agents to the reaction system.


Combined effects of cumulative stress and daily stressors on daily health.
It has been proposed that cumulative stress, one's experience of chronic stressors across multiple domains, worsens health by altering the extent to which daily stressors impact daily affect and physical symptoms. Recent work confirms that high cumulative stress exacerbates the association between daily stressor exposure and increased daily negative affect, though it remains untested the extent to which cumulative stress and daily stressor exposure interact to predict daily symptoms. We employed data from the second wave of the midlife in the U.S. Survey (N = 2,022; Mage = 56.2; 57.2% female) to examine whether levels of cumulative stress compound daily symptoms on days with (vs. without) stressful events. Experiences of life stressors across eight domains, occurrence of daily stressors, and occurrence, number, and severity of daily physical symptoms were analyzed using multilevel modeling. Greater cumulative stress and experiencing (vs. not experiencing) a daily stressor independently increased the odds of occurrence, number, and severity of daily symptoms (ps ≤ .016). Moreover, after adjusting for covariates (e.g., sociodemographic characteristics, chronic health conditions, percent of days with reported stressors, and health behaviors), the associations between daily stressor exposure and odds of occurrence, number, and severity of daily symptoms were potentiated as levels of cumulative stress increased (ps ≤ .009). The negative implications of daily stressor exposure for daily health may be most pronounced in those who report higher levels of cumulative stress across multiple life domains and across time. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


[Surveillance of nosocomial infections: prospective study in a pediatric intensive care unit. Background, patients and methods].
BACKGROUND, PATIENTS AND METHODS: From November 1997 through May 1998, the incidence of nosocomial infections was studied prospectively in a 10-bed multidisciplinary pediatric intensive care unit in Germany. A standardized surveillance [SEKI] system based on the National Nosocomial Infection Surveillance [NNIS] System of the Centers for Disease Control and Prevention [CDC] was used. The CDC definitions for nosocomial infections were adapted to the current practice of pediatric intensive care in Germany. Infection rates were calculated as infections per 100 patients, per 1000 patient-days, and per 1000 device-days (central venous catheters, urinary-catheters, and mechanical ventilation). Fifteen nosocomial infections were recorded in 201 patients during 1035 patient-days. The overall nosocomial infection rates were 7.5/100 patients and 14.5/1000 patient-days. Device-associated nosocomial infection rates for urinary-catheters and mechanical ventilation were 7.2/1000 utilization-days and thus below the 75th percentile of the last NNIS report. Central line infection rates were 10.7/1000 utilization days and therefore above the 75th percentile of the NNIS data (10.2/1000). The median length-of-stay was 5.1 days. Surveillance data are indispensable for internal and external quality control, and prospective surveillance of nosocomial infections should become an essential component of hospital infection control programs in pediatric intensive care in Germany. The standardized calculation of (device utilization ratios and) device-specific infection rates yields results which can be compared with national and international surveillance data. SEKI meets the criteria of a practice oriented, prospective and standardized surveillance system. Considerable efforts for collecting and interpreting the required data should be balanced against the benefit of prevention of nosocomial infections in this population of critically ill persons.


[DNA-cholesterol complex studied by fluorescent probes].
DNA interaction with cholesterol at various lipid concentrations has been investigated by the fluorescent probes method. It has been shown that the intensity of acridine orange fluorescence in the DNA-cholesterol complex decreases at 24 micrograms/ml cholesterol and at 45 micrograms/ml it increases. The number of binding sites and the degree of polarization of fluorescence change simultaneously. Binary mechanism of cholesterol binding with DNA has been suggested: surface binding takes place at low concentrations, intercalation--at high lipid concentrations.


Regulation of tumor immune surveillance and tumor immune subversion by tgf-Beta.
Transforming growth factor-beta (TGF-beta) is a highly pleiotropic cytokine playing pivotal roles in immune regulation. TGF-beta facilitates tumor cell survival and metastasis by targeting multiple cellular components. Focusing on its immunosuppressive functions, TGF-beta antagonists have been employed for cancer treatment to enhance tumor immunity. TGF-beta antagonists exert anti-tumor effects through #1 activating effector cells such as NK cells and cytotoxic CD8(+) T cells (CTLs), #2 inhibiting regulatory/suppressor cell populations, #3 making tumor cells visible to immune cells, #4 inhibiting the production of tumor growth factors. This review focuses on the effect of TGF-beta on T cells, which are differentiated into effector T cells or newly identified tumor-supporting T cells.


An evaluation of a C-reactive protein assay using a rate immunonephelometric procedure.
By using 20 repetitive assays at three levels of C-reactive protein control serum, within-run and day-to-day precision of a rate immunonephelometric system was established. The precision was excellent in each case. Linearity of the system between 1.8 and 20 mg/dl was established at a coefficient of correlation of 0.999 by using serial dilutions of C-reactive protein control serum. When the immunonephelometric assay was compared with electroimmunodiffusion and radial immunodiffusion assays, coefficients of correlation of 0.986 and 0.993, respectively, were obtained. The lower limit of sensitivity of the immunonephelometric assay was 1.8 mg/dl. The relationship of clinical significance to the performance of the C-reactive protein test system using rate immunonephelometry is discussed.


PGC-1alpha induces apoptosis in human epithelial ovarian cancer cells through a PPARgamma-dependent pathway.
Peroxisome proliferator-activated receptor gamma (PPARgamma) coactivator-1 alpha (PGC-1alpha) coactivates multiple transcription factors and regulates several metabolic processes. The current study investigated the role of PGC-1alpha in the induction of apoptosis in human epithelial ovarian cancer cells. The PGC-1alpha mRNA level between human ovaries and human ovarian epithelial tumors was examined by quantitative RT-PCR. Less PGC-1alpha expression was found in the surface epithelium of malignant tumors compared with normal ovaries. Overexpression of PGC-1alpha in human epithelial ovarian cancer cell line Ho-8910 induced cell apoptosis through the coordinated regulation of Bcl-2 and Bax expression. Microarray analyses confirmed that PGC-1alpha dramatically affected the apoptosis-related genes in Ho-8910 cells. Mitochondrial functional assay showed that the induction of apoptosis was through the terminal stage by the release of cytochrome c. Furthermore, PGC-1alpha-induced apoptosis was partially, but not completely, blocked by PPARgamma antagonist (GW9662), and suppression of PPARgamma expression by siRNA also inhibited PGC-1alpha-induced apoptosis in Ho-8910 cells. These data suggested that PGC-1alpha exerted its effect through a PPARgamma-dependent pathway. Our findings indicated that PGC-1alpha was involved in the apoptotic signal transduction pathways and downregulation of PGC-1alpha may be a key point in promoting epithelial ovarian cancer growth and progression.


Cellular mechanisms of burst firing-mediated long-term depression in rat neocortical pyramidal cells.
During wakefulness and sleep, neurons in the neocortex emit action potentials tonically or in rhythmic bursts, respectively. However, the role of synchronized discharge patterns is largely unknown. We have recently shown that pairings of excitatory postsynaptic potentials (EPSPs) and action potential bursts or single spikes lead to long-term depression (burst-LTD) or long-term potentiation, respectively. In this study, we elucidate the cellular mechanisms of burst-LTD and characterize its functional properties. Whole-cell patch-clamp recordings were obtained from layer V pyramidal cells in somatosensory cortex of juvenile rats in vitro and composite EPSPs and EPSCs were evoked extracellularly in layers II/III. Repetitive burst-pairings led to a long-lasting depression of EPSPs and EPSCs that was blocked by inhibitors of metabotropic glutamate group 1 receptors, phospholipase C, protein kinase C (PKC) and calcium release from the endoplasmic reticulum, and that required an intact machinery for endocytosis. Thus, burst-LTD is induced via a Ca2+- and phosphatidylinositol-dependent activation of PKC and expressed through phosphorylation-triggered endocytosis of AMPA receptors. Functionally, burst-LTD is inversely related to EPSP size and bursts dominate single spikes in determining the sign of synaptic plasticity. Thus burst-firing constitutes a signal by which coincident synaptic inputs are proportionally downsized. Overall, our data thus suggest a mechanism by which synaptic weights can be reconfigured during non-rapid eye movement sleep.


Dietary supplementation with highly purified eicosapentaenoic acid and docosahexaenoic acid does not influence PAI-1 activity.
Impaired fibrinolysis due to elevated levels of plasminogen activator inhibitor type 1 (PAI-1) is a risk factor for atherothrombotic disease. Many studies have reported a positive correlation between serum triglycerides and PAI-1 activity. Dietary intervention with very long n-3 fatty acids from marine sources is known to decrease serum triglycerides, but an adverse increase in PAI-1 activity has been reported in some studies. A double blind, placebo controlled study was conducted among 224 middle-aged (ages 36-56), healthy, non-smoking men in which the participants were randomly assigned to daily supplementation with 3.8 g eicosapentaenoic acid/d, 3.6 g docosahexaenoic acid/d, or 4.0 g corn oil/d (placebo) for 7 weeks. PAI-1 activity increased by 2.35+/-6.24 U/ml (28%), 1.15+/-6.74 U/ml (14%), and 1.33+/-5.64 U/ml (22%) during dietary supplementation with eicosapentaenoic acid, docosahexaenoic acid, and corn oil, respectively, but the changes were not significantly different between groups. There was no relationship between change in concentrations of serum triglycerides or phospholipid n-3 fatty acids and change in PAI-1 activity. At baseline, analysis was performed to investigate the influence of dietary lipids, blood lipids, and serum fatty acids on plasma concentrations of PAI-1 activity. Dietary intake of saturated fat correlated directly with PAI-1 both in crude analysis and after adjustment for age and body mass index (kg/m(2)). Furthermore, PAI-1 was associated with body mass index, apo-B100, serum triglycerides, and the concentration of n-6 polyunsaturated fatty acids in serum. In a multiple regression analysis, 21% of the variation in PAI-1 activity could be explained by these variables. Plasma PAI-1 activity did not correlate with dietary intake or serum concentrations of n-3 polyunsaturated fatty acids. In a review of 17 trials, including 935 subjects that assessed the effect of n-3 fatty acids on PAI-1 activity, an overall 17.7% increase in PAI-1 activity was estimated by n-3 supplementation. However, only two studies were able to demonstrate a significant increase in PAI-1 attributable to n-3 fatty acid supplementation. We conclude that there is no strong evidence for an unfavourable, clinically relevant effect of n-3 fatty acids on PAI-1 activity in plasma.


Association of amino acid variants in the activating transcription factor 6 gene (ATF6) on 1q21-q23 with type 2 diabetes in Pima Indians.
Activating transcription factor 6 (ATF6) is important for protective cell response to accumulation of unfolded and misfolded proteins in endoplasmic reticulum, and disturbances of this process can contribute to beta-cell apoptosis. We analyzed the structural gene located within a region on 1q21-q23 linked with type 2 diabetes in several populations for variants in the Pima Indians. Functionally important segments of ATF6 were sequenced in 15 diabetic and 15 nondiabetic Pimas and representative single nucleotide polymorphisms (SNPs) tested for association with type 2 diabetes in 900-1,000 subjects. We identified 20 variants including three amino acid substitutions [Met(67)Val, Pro(145)Ala, and Ser(157)Pro]. Pro(145)Ala and Ser(157)Pro were in a complete linkage disequilibrium and showed a nominal association with type 2 diabetes (P = 0.05; odds ratio 2.3 [95% CI 1.0-5.2]) and with 30-min plasma insulin during oral glucose tolerance test in 287 nondiabetic individuals (P = 0.045). Although the associations with type 2 diabetes and plasma insulin levels are marginal and their functional consequences are yet unknown, all three amino acid substitutions are located in a functionally important part of ATF6. Because these variants are not unique to the Pimas, it will be feasible to investigate their association with type 2 diabetes in other populations to better evaluate their significance for a predisposition to the disease.


Embryonic stem cell research: the state of the union of "those who create the lines" and "those who draw the lines".
Recent events have ostensibly removed the primacy of the United States in embryonic stem (ES) cell research. George Walker Bush was the first U.S. President to allow federal funding of ES cell research under guidelines influenced by statements from pre-eminent stem cell scientists. The guidelines delimiting federal funding of ES cell research put forth by President Bush were endorsed by the public, the National Institutes of Health (NIH) and by, at least by silent assent, the scientific community. At the time, the President was assured by eminent scientists and advisors that ES cells were immortal and stable, many viable lines existed, and they could be used for years for research and therapy. These premises have now been publicly challenged and individual states and other countries have announced plans to proceed with research that would be considered outside of the President's guidelines. All have thus far discovered that removal of U.S. federal support and oversight not only blunts science, but also abdicates responsible stewardship of the scientific process as well as the resulting technology. So while California, Canada, New Jersey, and Britain engage in ES research, suddenly the process stops; and no advances are made. Below I present a historical account of the seminal events and statements of legislators and ES cell researchers leading to and from that decision. The Journal welcomes our readers' comments on the issue at hand.


Industry Support of Patient Advocacy Organizations: The Case for an Extension of the Sunshine Act Provisions of the Affordable Care Act.
Patient advocacy organizations (PAOs) have long been regarded as important representatives of patient and caregiver interests in health policy debates. Recently, however, PAOs have attracted increased scrutiny over their financial ties to drug and device companies. In the past year, researchers and policymakers have called for the creation of a "sunshine law" requiring mandatory public reporting of industry payments to PAOs. Others have suggested that increased transparency would do little to address, and may even exacerbate, underlying concerns about proindustry bias among industry-funded PAOs. To date, however, the benefits of a sunshine law have not been well articulated, nor have objections to the idea been carefully addressed. In particular, little attention has been paid to clarifying the merits of statutorily mandated disclosure relative to those of increased voluntary disclosure by PAOs. I examine arguments for and against a sunshine law and conclude that the balance of reasons supports the enactment of such a law.


[Stabilization of colouring of Castellani's solution without fuchsin CL 2002 and control of its efficacy].
Disodium edetate in an amount of at least 0.01% is an effective stabilizer of a low level of colouring of Castellani's solution without fuchsin Ph.B. MMII. Spectrophotometric measurement of colouring and expressing in numbers of colour differences deltaE* in the uniform colour space CIELAB from transmittance seem to be a suitable method of stabilization efficacy control of colouring of the preparation.


Delayed axonal degeneration in slow Wallerian degeneration mutant mice detected using diffusion tensor imaging.
Previous studies have shown the feasibility of using diffusion tensor imaging (DTI) as a noninvasive imaging modality to evaluate neurodegeneration in humans and animals. The axial and radial diffusivities derived from DTI were demonstrated to be sensitive markers for axonal and myelin damage, respectively. This study used DTI to evaluate optic nerve degeneration in wild-type and slow Wallerian degeneration (Wld(S)) mutant mice. Longitudinal DTI was performed on optic nerves following high intraocular pressure-induced transient retinal ischemia. The axial diffusivity of wild-type nerves decreased 30% (P<0.05) at 3 days and 40% (P<0.05) at 5-30 days after transient elevation of intraocular pressure. In contrast, the axial diffusivity of Wld(S) nerves did not change at 3 days; decreased by 20% (P<0.05) at 5 days, and continued to decrease by 30% (P<0.05) at 15 days and 40% (P<0.05) at 30 days after transient intraocular pressure elevation, suggesting delayed axonal damage in Wld(S) mice. Radial diffusivity increased 200% (P<0.05) at 15-30 days in the wild-type mice and 100% (P<0.05) at 30 days in the Wld(S) mice after transient intraocular pressure elevation, suggesting delayed myelin damage in Wld(S) mice. DTI detected damage was confirmed with immunohistochemistry using phosphorylated neurofilament and myelin basic protein for assessing axonal and myelin integrity, respectively. These findings support the use of DTI not only to evaluate the progression of neurodegeneration but also to noninvasively demonstrate Wld(S) mutation to delay the Wallerian degeneration.


Coronary perfusion under general and selective cardiac hypothermia.
Experiments to assess left ventricular function in dogs during and after 2 hours of hypothermic cardioplegia are reported. During the experiment myocardial temperature was maintained between 15 degrees C and 18 degrees C by controlling the rate of flow of a cool cardioplegic solution into the coronary vessels. Before and after the period of hypothermia the left atrial pressure was gradually changed and the aortic flows and pressures that changed in response were recorded. The left ventricular stroke work (gm) was then calculated using the relationship: LVSM = (formula: see text) where: SV = stroke volume (ml), AP = arterial pressure (mmHg), LAP = left atrial pressure (mmHg). Comparison of the calculated values indicated that this method of maintaining cardiac hypothermia reduced cardiac function only slightly.


Physiologically Based Pharmacokinetic Modelling of Cabozantinib to Simulate Enterohepatic Recirculation, Drug-Drug Interaction with Rifampin and Liver Impairment.
Cabozantinib (CAB) is a receptor tyrosine kinase inhibitor approved for the treatment of several cancer types. Enterohepatic recirculation (EHC) of the substance is assumed but has not been further investigated yet. CAB is mainly metabolized via CYP3A4 and is susceptible for drug-drug interactions (DDI). The goal of this work was to develop a physiologically based pharmacokinetic (PBPK) model to investigate EHC, to simulate DDI with Rifampin and to simulate subjects with hepatic impairment. The model was established using PK-Sim® and six human clinical studies. The inclusion of an EHC process into the model led to the most accurate description of the pharmacokinetic behavior of CAB. The model was able to predict plasma concentrations with low bias and good precision. Ninety-seven percent of all simulated plasma concentrations fell within 2-fold of the corresponding concentration observed. Maximum plasma concentration (Cmax) and area under the curve (AUC) were predicted correctly (predicted/observed ratio of 0.9-1.2 for AUC and 0.8-1.1 for Cmax). DDI with Rifampin led to a reduction in predicted AUC by 77%. Several physiological parameters were adapted to simulate hepatic impairment correctly. This is the first CAB model used to simulate DDI with Rifampin and hepatic impairment including EHC, which can serve as a starting point for further simulations with regard to special populations.


[An in vitro study of lithium index in healthy Chinese].
The lithium index, a ratio of erythrocyte (intracellular) to plasma (extracellular) lithium concentrations, were studied by in vitro method in 22 healthy volunteers, 15 men and 17 women. Their mean age was 31.0 +/- 4.6 years (ranging from 26 to 41 years). In the study design, 20 ml of venous blood was taken from each subject into a test tube containing sodium citrate at the room temperature and erythrocytes were separated from plasma immediately. Then, the whole procedures for preparation and determination of lithium index were performed by using Pandey's method. Lithium levels were measured by atomic absorption spectrophotometry. Values of lithium index in vitro thus obtained were: 0.43 +/- 0.13 (range, 0.23-0.73) in men plus women; 0.37 +/- 0.10 (range, 0.23-0.57) in men; 0.49 +/- 0.13 (range, 0.26-0.73) in women. Women had a significantly higher Li+ index in vitro than men (t = 2.86, d.f. = 30, p less than 0.005). The possible reasons for the difference of the Li+ index in vitro between our result and other studies were discussed.


[Promotive excretion of causative agents of Yusho by fermented brown rice with Aspergillus oryze in Yusho patients].
Forty two years have passed since the outbreak of Kanemi rice oil poisoning, namely, Yusho in the western Japan. However, even now the Yusho patients have been still suffering from several objective and subjective symptoms. In order to improve or, if possible, to cure such symptoms, the most important therapeutic treatment is considered to actively excrete the causative agents, that is, polychlorinated dibenzofurans (PCDFs) from the bodies of the patients and to reduce their body burdens. In rats, chlorophyll and dietary fiber have been shown to promote the fecal excretion of PCDFs and polychlorinated dibenzo-p-dioxins (PCDDs) and to reduce their levels in rats. In this study, we have examined whether such kinds of effect are also observed by fermented brown rice with Aspergillus oryzae (FBRA) containing 5% spirulina, which is so-called spirulina HI * GENKI, the health food and relatively rich with chlorophyll and dietary fiber, in eighteen Yusho patients. They were divided into two groups, namely group A, ten patients (3 males and 7 females) with the mean age of 67.7 years old and group B, eight patients (4 males and 4 females) with the mean age of 64.1 years old. Among the patients of group A, three patients were especially highly contaminated with PCDFs and we classified them into group A (High). Respective mean concentrations of PCDFs in the blood just before initiating this study were as follows; group A : 145 pg/g lipid, group A (High) : 371 pg/g lipid and group B : 52.1 pg/g lipid. Contamination levels of PCDFs, PCDDs and dioxin-like polychlorinated biphenyls (PCBs) in group A (High) were 1.7 to 2.6 times higher than those in group A and 2.4 to 7.1 times higher than those in group B. Accordingly, concentrations of dioxins (PCDFs + PCDDs + dioxin-like PCBs) in the blood of groups A, A (High) and B were, respectively, 194, 458 and 85 pg-TEQ/g lipid. Concentrations of PCBs were also the highest in group A (High) : 1399 ng/g lipid, in group A : 748 ng/g lipid and the lowest in group B : 456 ng/g lipid. Groups A and A (High) took around 7.0 g of spirulina HI * GENKI after each meal and three times a day for the first one year and for the second one year, they did not take spirulina HI * GENKI anymore. Group B took spirulina HI * GENKI with the same manner as the group A only for the second one year. The concentrations of PCDFs, PCDDs, dioxin-like PCBs and PCBs in the blood were also measured at the end of the first and second year, respectively. Assuming the body fat is also contaminated with these compounds at their concentrations on lipid weight basis in the blood and the content of body fat is 20% of 60 kg body weight, we computed the average amounts in their net excretion from the body of the patients due to spirulina HI * GENKI in the three groups. As a result, in group A (High), 341 ng-TEQ/patient of dioxins was excreted from the body, which was 3.4 times greater than that in group A and 12 times higher than that in group B. Therefore, promotive excretions of causative agents of Yusho were the most effective in group A (High) and we consider spirulina HI * GENKI is more effective from the therapeutic point of view in more highly contaminated Yusho patients.


Identification of interacting transcription factors regulating tissue gene expression in human.
Tissue gene expression is generally regulated by multiple transcription factors (TFs). A major first step toward understanding how tissues achieve their specificity is to identify, at the genome scale, interacting TFs regulating gene expression in different tissues. Despite previous discoveries, the mechanisms that control tissue gene expression are not fully understood. We have integrated a function conservation approach, which is based on evolutionary conservation of biological function, and genes with highest expression level in human tissues to predict TF pairs controlling tissue gene expression. To this end, we have identified 2549 TF pairs associated with a certain tissue. To find interacting TFs controlling tissue gene expression in a broad spatial and temporal manner, we looked for TF pairs common to the same type of tissues and identified 379 such TF pairs, based on which TF-TF interaction networks were further built. We also found that tissue-specific TFs may play an important role in recruiting non-tissue-specific TFs to the TF-TF interaction network, offering the potential for coordinating and controlling tissue gene expression across a variety of conditions. The findings from this study indicate that tissue gene expression is regulated by large sets of interacting TFs either on the same promoter of a gene or through TF-TF interaction networks.


The challenge and importance of standardizing pre-analytical variables in surgical pathology specimens for clinical care and translational research.
The introduction of targeted cancer therapies into clinical practice, in which patients are selected for novel treatments based on results of companion molecular testing of their tumor specimens, has created significant new challenges for the surgical pathology laboratory. These include standardization of tissue handling and sample preparation with accurate documentation to ensure optimal quality of clinical samples to reduce the risk of errors in molecular biology tests. The assay of tumor tissues for biomarkers that can provide predictive data for prognosis or treatment should enable selection of the most appropriate therapies (Yaziji et al. 2008, Hicks and Kulkarni 2008). Major advances have been made in the ability to profile clinical samples for research at the DNA, RNA and protein levels. To translate this new information into the clinical setting, however, the quality of the starting material, in this case the tumor tissue, determines the accuracy and reliability of companion diagnostic assay results and therefore optimal therapeutic strategies. Inaccurate results owing to compromised tissue quality can lead to false positive or false negative results with therapeutic consequences that can harm patients and affect their eventual outcome.


Pulmonary blood volume in mitral regurgitation in Cavalier King Charles spaniels.
Pulmonary edema and venous congestion are well-recognized signs of congestive heart failure (CHF) in advanced canine chronic mitral regurgitation (MR). However, little is known about pulmonary blood volume (PBV), blood pulmonary transit time (PTT), and the regulation of these. To measure and evaluate the relationships of PBV, forward stroke volume (FSV), and heart rate normalized blood pulmonary transit time (nPTT) in healthy dogs and dogs with MR. Thirty-three Cavalier King Charles Spaniels; 11 healthy, 4 in modified New York Heart Association (NYHA) class I, 11 in class II, and 7 in CHF. Heart rate normalized PTTs were measured by radionuclide angiocardiography. Left ventricular end diastolic and systolic diameter, left atrial/aortic root ratio, and FSV were measured by echocardiography. PBV and pulmonary blood volume index (PBVI) were calculated by established formulas. PBVI was 308±56 (mean±SD) mL/m2 for healthy dogs, 287±51 mL/m2 in NYHA class I, 360±66 mL/m2 in Class II, and 623±232 mL/m2 in CHF (P=.0008). Heart rate normalized PTT, not FSV, was a predictor of PBV (r=0.92 and 0.02, respectively). Increased PBV, not decreased FSV, is the main cause of increased nPTT in MR. Increased nPTT can be used as an indicator of abnormal cardiopulmonary function in dogs with MR.


Light and temperature regulate m6A-RNA modification to regulate growth in plants.
N6-methyladenosine is a highly dynamic, abundant mRNA modification which is an excellent potential mechanism for fine tuning gene expression. Plants adapt to their surrounding light and temperature environment using complex gene regulatory networks. The role of m6A in controlling gene expression in response to variable environmental conditions has so far been unexplored. Here, we map the transcriptome-wide m6A landscape under various light and temperature environments. Identified m6A-modifications show a highly specific spatial distribution along transcripts with enrichment occurring in 5'UTR regions and around transcriptional end sites. We show that the position of m6A modifications on transcripts might influence cellular transcript localization and the presence of m6A-modifications is associated with alternative polyadenylation, a process which results in multiple RNA isoforms with varying 3'UTR lengths. RNA with m6A-modifications exhibit a higher preference for shorter 3'UTRs. These shorter 3'UTR regions might directly influence transcript abundance and localization by including or excluding cis-regulatory elements. We propose that environmental stimuli might change the m6A landscape of plants as one possible way of fine tuning gene regulation through alternative polyadenylation and transcript localization.


Development and Optimization of Cold Plasma Pretreatment for Drying on Corn Kernels.
This study was conducted to optimize a process of cold plasma pretreatment for hot-air drying on corn kernels. Effects of plasma pretreatment time (30, 40, 50 s), plasma pretreatment power (300, 400, 500 W) and drying temperature (37.5, 45, 52.5 °C) on drying time and drying rate during this process were investigated. Polynomial equations were established through a three-factor and three-level Box-Behnken design and used to evaluate the optimal operational conditions for the drying process. The optimal pretreatment conditions were drying temperature at 52.5 °C, plasma pretreatment time of 50 s and plasma pretreatment power of 500 W, and the corresponding drying rate and drying time were 3.6163 (g/g h-1 ) and 1.29 hr, respectively. The AFM images showed that cold plasma pretreatment can change the topography of the treated surface with some micro-holes, which explain how the plasma pretreatment can improve the drying process. PRACTICAL APPLICATION: Cold plasma pretreatment can improve the efficiency of corn kernels drying. Furthermore, it has potential application for reducing energy consumption in drying. Cold plasma pretreatment could be potentially applied in grain drying.


Protein phosphatase-1 modulates the function of Pax-6, a transcription factor controlling brain and eye development.
Pax-6 is an evolutionarily conserved transcription factor and acts high up in the regulatory hierarchy controlling eye and brain development in humans, mice, zebrafish, and Drosophila. Previous studies have shown that Pax-6 is a phosphoprotein, and its phosphorylation by ERK, p38, and homeodomain-interacting protein kinase 2 greatly enhances its transactivation activity. However, the protein phosphatases responsible for the dephosphorylation of Pax-6 remain unknown. Here, we present both in vitro and in vivo evidence to show that protein serine/threonine phosphatase-1 is a major phosphatase that directly dephosphorylates Pax-6. First, purified protein phosphatase-1 directly dephosphorylates Pax-6 in vitro. Second, immunoprecipitation-linked Western blot revealed that both protein phosphatase-1alpha and protein phosphatase-1beta interact with Pax-6. Third, overexpression of protein phosphatase-1 in human lens epithelial cells leads to dephosphorylation of Pax-6. Finally, inhibition of protein phosphatase-1 activity by calyculin A or knockdown of protein phosphatase-1alpha and protein phosphatase-1beta by RNA interference leads to enhanced phosphorylation of Pax-6. Moreover, our results also demonstrate that dephosphorylation of Pax-6 by protein phosphatase-1 significantly modulates its function in regulating expression of both exogenous and endogenous genes. These results demonstrate that protein phosphatase 1 acts as a major phosphatase to dephosphorylate Pax-6 and modulate its function.


Hemodynamic and electrophysiological effects of a novel positive inotropic drug, OPC-8212, in normal and "failing" guinea pig heart preparations.
OPC-8212, 3,4-dihydro-6-[4-(3,4-dimethoxy-benzoyl)-1-piperazinyl]-2 (1H)-quinolinone, significantly decreased heart rate and increased contractility (+ dP/dt) and coronary flow in isolated, perfused guinea pig Langendorff and work-performing heart preparations. The effect on contractility, but not on coronary flow, was significantly greater when the negative chronotropic effect was prevented by pacing. In guinea pig hearts made incompetent by aortic stenosis, OPC-8212 produced a significant increase in contractility and coronary flow and greater negative chronotropic effects than in control hearts. OPC-8212 improved the work performance of normal and "failing" isolated work-performing guinea pig hearts during pressure load and during volume load. In electrophysiological studies, OPC-8212 enhance Vmax, amplitude, and duration of fast action potentials (APs) in guinea pig papillary muscles. Tetraethylammonium (TEA)-induced slow APs were also potentiated with respect to Vmax, amplitude, and duration. OPC-8212 in the absence of TEA also lowered the voltage threshold for inducing the slow APs. Positive inotropic effects, but not the slow APs, of guinea pig papillary muscles were greatly enhanced by OPC-8212 at higher frequencies of stimulation (2-3 Hz), indicating an important mechanism of action that may be at least in part independent of Isi.


Streptomyces chiangmaiensis SSUT88A mediated green synthesis of silver nanoparticles: characterization and evaluation of antibacterial action against clinical drug-resistant strains.
This study involved the characterization of AgNPs synthesized from soil isolate Streptomyces sp. SSUT88A and their antimicrobial activities. The strain SSUT88A revealed 98.8% similarity of the 16s rRNA gene to Streptomyces chiangmaiensis TA4-1T. The AgNPs were synthesized by mixing either intracellular or extracellular cell-free supernatant of strain SSUT88A with AgNO3. The synthesized AgNPs from intracellular cell-free supernatant and extracellular cell-free supernatant were designated as IS-AgNPs and ES-AgNPs, respectively. The IS-AgNPs showed maximum absorbance of UV-vis spectra at 418 nm, while ES-AgNPs revealed maximum absorbance at 422 nm. The TEM observation of synthesized AgNPs revealed a spherical shape with an average diameter of 13.57 nm for IS-AgNPs and 30.47 nm for ES-AgNPs. The XRD and XANES spectrum profile of both synthesized AgNPs exhibited similar spectrum energy, which corresponded to AgNPs. The IS-AgNPs revealed antimicrobial activity against clinical isolate drug-resistant bacteria (Acinetobacter baumannii, Escherichia coli 8465, Klebsiella pneumoniae 1617, and Pseudomonas aeruginosa N90PS), while ES-AgNPs had no antimicrobial activity. When compared to commercial AgNPs, IS-AgNPs exhibited antibacterial efficacy against all clinical isolate bacteria including A. baumannii, one of the most threatening multi-drug resistant strains, while commercial AgNPs did not. Thus, IS-AgNPs has potential to be further developed as an antimicrobial agent against drug-resistant bacteria.


Addressing Disparities in Physician Access to Information in Support of Evidence-based Practice.
The purpose of this study is to determine if Minnesota physicians have access to information resources needed to support evidence-based practice (EBP), which supports a culture of safety and patient-centered care. A survey was used to determine Minnesota physicians' need for, and access to, evidence-based clinical information. A total of 877 responses (6.4% response rate) were included in the data analysis. Participants spent 24 min daily seeking answers to clinical questions and averaged 4.41 questions per day that could not be immediately answered. Physicians reported high levels of information needs met (85.8%), though they reported limited access to drug resources, citation databases, systematic reviews, and full-text books and articles. Results also showed use of unreliable sources to support decision-making. A key finding was the extent to which workplace affiliation broadens disparities in information access. National and regional approaches can work to support EBP by reducing the information gap caused by workplace affiliation and other barriers. Further research should be done to identify partnerships, funding, infrastructure, and support to address these gaps.


Stretchable Capacitive Pressure Sensing Sleeve Deployable onto Catheter Balloons towards Continuous Intra-Abdominal Pressure Monitoring.
Intra-abdominal pressure (IAP) is closely correlated with intra-abdominal hypertension (IAH) and abdominal compartment syndrome (ACS) diagnoses, indicating the need for continuous monitoring. Early intervention for IAH and ACS has been proven to reduce the rate of morbidity. However, the current IAP monitoring method is a tedious process with a long calibration time for a single time point measurement. Thus, there is the need for an efficient and continuous way of measuring IAP. Herein, a stretchable capacitive pressure sensor with controlled microstructures embedded into a cylindrical elastomeric mold, fabricated as a pressure sensing sleeve, is presented. The sensing sleeve can be readily deployed onto intrabody catheter balloons for pressure measurement at the site. The thin and highly conformable nature of the pressure sensing sleeve captures the pressure change without hindering the functionality of the foley catheter balloon.


Autonomous activity of CaMKII is only transiently increased following the induction of long-term potentiation in the rat hippocampus.
A major role has been postulated for a maintained increase in the autonomous activity of CaMKII in the expression of long-term potentiation (LTP). However, attempts to inhibit the expression of LTP with CaMKII inhibitors have yielded inconsistent results. Here we compare the changes in CaMKII autonomous activity and phosphorylation at Thr286 of alphaCaMKII in rat hippocampal slices using chemical or tetanic stimulation to produce either LTP or short-term potentiation (STP). Tetanus-induced LTP in area CA1 requires CaMKII activation and Thr286 phosphorylation of alphaCaMKII, but we did not observe an increase in autonomous activity. Next we induced LTP by 10 min exposure to 25 mM tetraethyl-ammonium (TEA) or 5 min exposure to 41 mM potassium (K) after pretreatment with calyculin A. Exposure to K alone produced STP. These protocols allowed us to monitor temporal changes in autonomous activity during and after exposure to the potentiating chemical stimulus. In chemically induced LTP, autonomous activity was maximally increased within 30 s whereas this increase was significantly delayed in STP. However, in both LTP and STP the two-fold increase in autonomous activity measured immediately after stimulation was short-lived, returning to baseline within 2-5 min after re-exposure to normal ACSF. In LTP, but not in STP, the phosphorylation of alphaCaMKII at Thr286 persisted for at least 60 min after stimulation. These results confirm that LTP is associated with a maintained increase in autophosphorylation at Thr286 but indicate that a persistent increase in the autonomous activity of CaMKII is not required for the expression of LTP.


Vitamin D-mediated hypercalcemia in lymphoma: evidence for hormone production by tumor-adjacent macrophages.
Nearly one-half of all hypercalcemic patients with lymphoma present with inappropriately elevated circulating concentrations of the active vitamin D metabolite 1,25-dihydroxyvitamin D (1,25(OH)2D3). However, the cellular source of the vitamin D hormone in lymphomas remains unclear. To address this, we report the case of a 75-year-old man with hypercalcemia associated with raised circulating concentrations of 1,25(OH)2D3 and suppressed parathyroid hormone (PTH) levels. Positron emission tomographic (PET) and computed tomographic (CT) imaging revealed the presence of a large lymphoma that was confined to the spleen; subsequent pathological analysis showed that this was an intermediate grade B-cell lymphoma. After surgical removal of the spleen, serum calcium and 1,25(OH)2D3 levels became normalized within 24 h. Immunolocalization of the vitamin D-activating enzyme 25-hydroxyvitamin D3-1alpha-hydroxylase (la-hydroxylase) in sections of resected spleen showed that staining was negative in the lymphoma cells but positive in neighboring macrophages. This case study indicates that the hypercalcemia associated with lymphomas may be due, in some instances, to excessive extrarenal production of 1,25(OH)2D3. Furthermore, by using immunohistochemistry to assess the distribution of la-hydroxylase, we have been able to show for the first time that tissue macrophages, rather than actual tumor cells, are the most likely ectopic source of this enzyme. Based on this case study, we propose that the abnormal synthesis of 1,25(OH)2D3 associated with some lymphomas is because of paracrine regulation of tumor-associated macrophages.


Increasing the antioxidant content of food: a personal view on whether this is possible or desirable.
A commonly held belief is that higher intake of antioxidants will promote better health through enhanced antioxidant status and lowered oxidative stress. However, the benefits of antioxidant-rich foods have not been reproduced in supplementation trials with pure antioxidants. This has driven research and commercial interest in foods, including traditional foods and their components, with enhanced antioxidant content and improved antioxidant bioavailability, which in many cases is very low. In this paper, evidence for the health benefits of antioxidant-rich foods and methods to increase the antioxidant content and bioavailability of food antioxidants are reviewed briefly, and the concept that increased food antioxidant content/intake per se is beneficial is examined from a cautionary perspective, considering issues of low bioavailability, rapid catabolism, biotransformation and the paradoxical pro-oxidant effects of dietary antioxidants.


Indium-sulfur supertetrahedral polymers integrated with [M(phen)3]2+ cations (M = Ni and Fe).
A series of supertetrahedral polymers of chalcogenometalates (T3 cluster compounds) integrated with M-phen complexes (phen =1,10-phenanthroline; M = Ni, Fe) was prepared by a similar solvothermal technique. Compound [Fe(phen)(3)](4)[H(4)In(20)S(38)]·Hphen·3HDMA·8H(2)O (Mp-InS-4) (DMA = dimethylamine) is a 1-D straight chain. Compounds [M(phen)(3)](4)[In(20)S(37)]·6Hphen·4H(2)O (M = Ni, Mp-InS-5; Fe, Mp-InS-6) are the first reported 2-D Tn polymers integrated with complex cations of [M(phen)(3)](2+). Compound [Ni(phen)(3)](4)[H(4)In(20)S(38)]·2Hphen·2HDMA·3H(2)O (Mp-InS-7) shows a zigzag 1-D structure. We find that the reaction time is an important factor in assembling of the T3 clusters. Prolonging the reaction time seems favorable to the higher condensed phases (from 0-D to 2-D). However, a longer reaction time resulted in the crack of 2-D structure. Integrating M-phen complex cations with the chalcogenido anions can improve absorption of the materials in the visible range due to the charge transfers within the cations or between cations and anions.


Cytotactin, an extracellular matrix protein of neural and non-neural tissues that mediates glia-neuron interaction.
An extracellular matrix protein, cytotactin, with widespread tissue distribution has been identified, isolated, and partially characterized. Cytotactin mediates glia-neuron adhesion in vitro, but unlike Ng-CAM, the neuron-glia cell-adhesion molecule, it is absent from neurons. Cytotactin was isolated from 14-day embryonic chicken brains as structurally related polypeptides of Mr 220,000, 200,000, and 190,000. These polypeptides were efficiently extracted in the absence of detergent and appeared to be disulfide-linked into higher polymers. Immunofluorescence staining with specific antibodies indicated that cytotactin is found in extracellular spaces and in basement membranes of a variety of non-neural tissues including smooth muscle, lung, and kidney. In the cerebellum, it appears on glial end-feet, on Bergmann glial fibers, and in extracellular spaces. The molecule is synthesized by glia and cells from smooth muscle, lung, and kidney. It is found at the surface of glia in culture in a cell-associated fibrillar pattern. A survey of the times and sites of its appearance during embryogenesis is consistent with the hypothesis that cytotactin is a cell-substrate adhesion molecule that may mediate cell migration in a site-restricted fashion.


Comparative Physiological and Transcriptomic Analyses of Improved Heat Stress Tolerance in Celery (Apium Graveolens L.) Caused by Exogenous Melatonin.
Melatonin (MT) is crucial in plant growth, development, and response to stress. Celery is a vegetable that grows in a cool climate, and a hot climate can deteriorate its growth, yield, and quality. This study investigates the effect of exogenous melatonin on celery physiology. Transcriptional levels were analyzed by spraying celery with exogenous MT before exposing it to high temperatures. The regulatory mechanism of exogenous MT-mediated heat tolerance was examined. The results show that the exogenous MT reduced the thermal damage state of celery seedlings, as well as the malondialdehyde (MDA) content and relative conductivity (REC), increasing the oxidase activity, the osmotic regulatory substances, and chlorophyll, enhancing the leaf transpiration and the light energy utilization efficiency. We examined the mechanism of exogenous MT in mitigating high-temperature damage using the transcriptome sequencing method. A total of 134 genes were expressed differently at high temperature in the celery treated with MT compared with the untreated celery. Functional annotation analysis revealed that the differentially expressed genes were abundant in the "pyruvate metabolism" pathway and the "peroxidase activity" pathway. According to the pathway-based gene expression analysis, exogenous MT can inhibit the upregulation of pyruvate synthesis genes and the downregulation of pyruvate consumption genes, preventing the accumulated pyruvate from rapidly upregulating the expression of peroxidase genes, and thereby enhancing peroxidase activity. RT-qPCR verification showed a rising encoding peroxidase gene expression under MT treatment. The gene expression pattern involved in pyruvate anabolism and metabolism agreed with the abundant transcriptome expression, validating the physiological index results. These results indicate that the application of exogenous MT to celery significantly enhances the ability of plant to remove reactive oxygen species (ROS) in response to heat stress, thereby improving the ability of plant to resist heat stress. The results of this study provide a theoretical basis for the use of MT to alleviate the damage caused by heat stress in plant growth and development.


[A case of sensory perineuritis with Bowen disease].
A 60-year-old man was admitted because of subacutely progressive sensory disturbance characterized by fluctuating dysesthesia and light pain on the skin of his hands and feet. He exhibited mononeuritis multiplex, and a plaque with sloughing was observed on the left side of his back. Sural nerve biopsy revealed marked thickening of the perineurium with vascularization, and no necrotizing vasculitis. The pathological features of the nerve biopsy were compatible with sensory perineuritis. A biopsy from his skin lesion revealed Bowen's disease. There have been several reported cases of sensory perineuritis accompanied by malignant tumors. In these cases, immune dysfunctions were considered a common underlying cause in both diseases. This is the first reported case of sensory perineuritis associated with Bowen's disease.


Production of progesterone from de novo-synthesized cholesterol in cumulus cells and its physiological role during meiotic resumption of porcine oocytes.
To investigate the role of factors secreted by cumulus cells during meiotic resumption of porcine oocytes, 1, 5, 10, or 20 cumulus-oocyte complexes (COCs) were cultured in each well of a culture dish containing 300 microl of maturation medium for 20 h. There was a significant positive correlation between the rate of germinal vesicle breakdown (GVBD) and the number of COCs cultured in each well for 20 h. The level of progesterone in the medium in which COCs had been cultured for 20 h also rose significantly with an increase in the number of COCs cultured in each well. A significantly small proportion of GVBD in oocytes when one COC was cultured in each well for 20 h was improved by the addition of progesterone. This proportion of GVBD was fully comparable to that of COCs cultured in the absence of additional progesterone with 20 COCs. Thus, progesterone secreted by COCs plays a positive role in GVBD induction in porcine oocytes. Furthermore, we also examined the role of sterol biosynthesis on progesterone production by cumulus cells and in oocyte GVBD. The results showed that the addition of ketoconazole, which suppressed the sterol biosynthetic pathway produced by demethylation of lanosterol, decreased the rate of GVBD, as well as progesterone production in COCs cultured for 20 h. However, the suppression of GVBD by ketoconazole was overtaken by the addition of progesterone. These results demonstrate that a high level of progesterone produced by cumulus cells was responsible for an acceleration of GVBD in porcine oocytes.


A conserved helical capping hydrogen bond in PAS domains controls signaling kinetics in the superfamily prototype photoactive yellow protein.
PAS domains form a divergent protein superfamily with more than 20 000 members that perform a wide array of sensing and regulatory functions in all three domains of life. Only nine residues are well-conserved in PAS domains, with an Asn residue at the start of α-helix 3 showing the strongest conservation. The molecular functions of these nine conserved residues are unknown. We use static and time-resolved visible and FTIR spectroscopy to investigate receptor activation in the photosensor photoactive yellow protein (PYP), a PAS domain prototype. The N43A and N43S mutants allow an investigation of the role of side-chain hydrogen bonding at this conserved position. The mutants exhibit a blue-shifted visible absorbance maximum and up-shifted chromophore pK(a). Disruption of the hydrogen bonds in N43A PYP causes both a reduction in protein stability and a 3400-fold increase in the lifetime of the signaling state of this photoreceptor. A significant part of this increase in lifetime can be attributed to the helical capping interaction of Asn43. This extends the known importance of helical capping for protein structure to regulating functional protein kinetics. A model for PYP activation has been proposed in which side-chain hydrogen bonding of Asn43 is critical for relaying light-induced conformational changes. However, FTIR spectroscopy shows that both Asn43 mutants retain full allosteric transmission of structural changes. Analysis of 30 available high-resolution structures of PAS domains reveals that the side-chain hydrogen bonding of residue 43 but not residue identity is highly conserved and suggests that its helical cap affects signaling kinetics in other PAS domains.


Separation of deformable particles in deterministic lateral displacement devices.
Using numerical simulations, we study the separation of deformable bodies, such as capsules, vesicles, and cells, in deterministic lateral displacement devices, also known as bump arrays. These arrays comprise regular rows of obstacles such as micropillars whose arrangements are shifted between adjacent rows by a fixed amount. We show that, in addition to the zigzag and laterally displaced trajectories that have been observed experimentally, there exists a third type of trajectory which we call dispersive, characterized by seemingly random bumpings off the micropillars. These dispersive trajectories are observed only for large and rigid particles whose diameters are approximately more than half the gap size between micropillars and whose stiffness exceeds approximately 500 MPa. We then map out the regions in phase space, spanned by the row shift, row separation, particle diameter, and particle deformability, in which the different types of trajectories are expected. We also show that, in this phase space, it is possible to transition from zigzag to dispersive trajectories, bypassing lateral displacement. Experimentally, this is undesirable because it limits the ability of the device to sort particles according to size. Finally, we discuss how our numerical simulations may be of use in device prototyping and optimization.


Expression and prognostic potential of TMEM204: a pan-cancer analysis.
TMEM204 (Transmembrane Protein 204) is a member of the TMEM family that regulates cell function and angiogenesis. Previous studies showed that TMEM204 is related to pancreatic cancer, but its roles in other cancers remain unknown. To reveal this relationship, we conducted a pan-cancer analysis by several online databases. The expression of TMEM204 was analyzed by Oncomine and Tumor Immune Estimation Resource2.0 (TIMER2.0). The prognostic potential of TMEM204 was evaluated by the GEPIA2, UALCAN, and Oncolnc. The methylation level of gene expression was analyzed by UALCAN, and the relationship between cancer and immune invasion was displayed by TIMER2.0. The Protein-Protein Interactions Network and functional analysis of TMEM204 and its related genes were conducted by STRING and Webgestalt. We found that TMEM204 expression was up-regulated and correlated with prognosis in multiple cancers. In liver hepatocellular carcinoma (LIHC), high TMEM204 expression was associated with a good prognosis, and with high infiltrating levels of CD8+ T and CD4+ T cells, macrophages, neutrophils, and myeloid dendritic cells. In addition, the methylation level in LIHC was higher than in normal tissues. p53 signaling pathway and Fanconi anemia pathway were implicated by KEGG pathway analysis. These results indicate that TMEM204 is associated with the prognosis, methylation, and immune invasion of cancers, especially LIHC. TMEM204 may act as a prognostic marker of LIHC and its role in other cancers should be studied.


Barriers to End-of-Life Care for LGBT Persons in the Absence of Legal Marriage or Adequate Legal Documentation.
End-of-life care has attracted increased attention in recent years due to increases in both the number and mean age of the world's population; however, the experiences of LGBT persons during end-of-life care remain understudied. Given the health disparities and barriers to care experienced throughout the life course of LGBT persons, the frequent involvement of legal spouses in end-of-life care, and the recency of marriage equality, it can be surmised that LGBT persons might experience significantly different barriers to their desired end-of-life care compared to their heterosexual and cisgender counterparts. This article aims to synthesize what is known about these barriers, particularly in the absence of legal marriage or protective legal documentation. Of two hundred and twelve articles reviewed, twenty-three were included for analysis. Common barriers that emerged were discriminatory laws (e.g., prohibitions against same-sex marriage) and policies, lack of decision-making capacity, lack of knowledge regarding patient wishes, lack of visitation rights, challenges from biological next of kin, and discrimination and psychological distress. Recommendations for future research are provided based on gaps that were identified. These include increased research on transgender persons and bisexual persons, and on providers of end-of-life care. In addition, the recommendations take into account that important legislative and policy changes occurred after the period in which the studies reviewed here were published. The impact of those changes cannot be reported in this study and should be determined by future studies. This synthesis provides the contextual understanding necessary for research and improvement in this vastly understudied area.


Return of autologous blood donors as homologous blood donors.
Autologous blood donors (ABDs) have been reported to have favorable attitudes toward returning as homologous blood donors (HBDs), but the frequency of return has not been well documented. ABDs eligible by history to be HBDs were followed at one blood center: 255 donating for elective surgery and 234 donating during pregnancy were followed for an average of 18 months and 20 months, respectively, from time of eligibility after surgery or postpartum. Male ABDs had a higher rate of return as HBDs, as 34 percent (21/62) returned to donate an average of 3 units, whereas 13 percent (56/427) of female ABDs returned as HBDs to donate an average of 2 units. Although a history of donation was associated with a higher rate of return (30%, 34/113), 11 percent (43/376) of ABDs with no history as HBDs returned to donate homologous units, despite having been recruited less frequently than prior HBDs. Overall, all male ABDs and female ABDs with an HBD history returned most frequently. The extra effort required for an autologous donor program may result in the recruitment of new donors into the HBD pool.


[Strategies of nutrients control in lakes based on ecoregions of lakes in China].
In order to better reduce lake eutrophication, based on five ecoregions of lakes, the relationships of total nitrogen (TN), total phosphorus (TP), and TN/TP ratio with chlorophyll-a (Chl-a) in 100 lakes were discussed, furthermore, strategies of nutrient control were proposed respectively. Results showed that among the five ecoregions, the lake eutrophication level was the lowest in Dongbei ecoregion and the highest in Huabei ecoregion, but the eutrophication level in lakes of all the five ecoregions is increasing in recent years. Algal growth in lakes of Dongbei and Huabei ecoregions was limited by phosphorus, while it was simultaneously limited by nitogen and phosphorus in lakes of other three ecoregions (Zhongdongbu, Yungui and Mengxin). In lakes with TN/TP < 10, significant correlation between Chl-a concentration and TN was found in lakes of the five ecoregions except for Huabei ecoregion, and significant correlation between Chl-a concentration and TP was found in lakes of Dongbei and Mengxin ecoregions. In lakes with TN/TP > 17, significant correlation between Chl-a concentration and TP was found in lakes of the five ecoregions except for Mengxin ecoregion, and significant correlation between Chl-a concentration and TN was found in lakes of Zhongdongbu, Yungui and Mengxin ecoregions. In lakes with 10 < TN/TP < 17, no significant correlation between Chl-a and TN (or TP) was found in lakes of all ecoregions except for Zhongdongbu ecoregion where Chl-a concentration was significantly affected by TN. As for strategies of nutrient control and reduction in the five ecoregions, lakes of Huabei ecoregion should adopt TP control in priority, and in Dongbei ecoregion, TP and TN should be controlled simultaneously in lakes with TN/TP < 10, while other lakes should focus on TP control. Lakes in other three ecoregions (Zhongdongbu, Mengxin and Yungui) should control TP and TN simultaneously.


Lesions of the putamen and dystonia: CT and magnetic resonance imaging.
We studied posthemiplegic hemidystonia in an adult, and generalized dystonia in two children. CT and magnetic resonance imaging (MRI) studies in the adult revealed infarction of the contralateral putamen and, to a much lesser extent, the head of the caudate nucleus. Both children had subacute encephalopathies (possible Leigh's disease), and CT revealed bilateral putamen lesions when generalized dystonia was the predominant clinical disorder. These cases and other reports of symptomatic dystonia suggest that lesions of the putamen correlate with dystonia.


Explicit Sequence Memory in Recall of Temporally-structured Episodes.
The order in which events unfold over time is an important scaffold aiding recollection. This study asks whether explicit order memory is enhanced for items sharing similar internally-driven temporal contexts. To tap internally-driven temporal context, we capitalized on the Temporal Contiguity Effect whereby recollection of one item promotes recall of adjacently-encoded items. We compared pairs encoded and retrieved contiguously (cont-enc-ret), whose items share internally-driven temporal contexts, to pairs retrieved, but not encoded, contiguously (cont-ret) and to pairs encoded, but not retrieved, contiguously (cont-enc). Cont-enc-ret pairs exhibited superior relative order over cont-ret pairs, supporting accounts emphasizing shared temporal context as opposed to temporal distinctiveness in driving sequence memory. No difference was found in absolute order between the pair types, in line with theories suggesting a dissociation between relative and absolute order. Additionally, cont-enc-ret and cont-enc pairs exhibited equivalent relative order, supporting the role of encoding as opposed to retrieval in the enhancement of relative order. Finally, cont-enc-ret pairs were perceived as closer than cont-enc pairs, supporting the claim that cont-enc-ret pairs constitute part of a temporally-coherent episode. Together, these results implicate internally-driven temporal context in the formation of temporally-structured episodes that enhances sequence memory of the items within the episode.


Interleukin 10 and tumor necrosis factor-alpha genotypes in rheumatoid arthritis--association with clinical response to glucocorticoids.
There are dysregulated levels of interleukin 10 (IL-10) and tumor necrosis factor-alpha (TNF-alpha) in rheumatoid arthritis (RA), and their role in the disease is controversial. We analyzed the association of functional polymorphisms of IL-10 and TNF-alpha with susceptibility and disease characteristics at the time of diagnosis, and we also evaluated their possible use as predictors of clinical response to treatments. Patients with recent-onset RA (n = 162) and healthy controls (n = 373) were genotyped for -1082 IL-10 and -308 TNF-alpha polymorphisms and data were related to clinical and immunological measurements of patients at the time of diagnosis. Response to treatment after 6 months was determined in 125 patients by the absolute change in Disease Activity Score (DAS28) and the American College of Rheumatology criteria for improvement. We found a reduced frequency of the low IL-10 producer genotype (-1082AA) in patients with RA compared to controls (26.5% vs 38.9%; p = 0.006), while it is a risk factor for anticyclic citrullinated peptide antibodies (anti-CCP) positivity (p = 0.028). Evaluation of clinical response to treatments indicated that carriage of the high IL-10 genotype was associated with a favorable outcome (p = 0.009), specifically to prednisone therapy (p = 0.0003). No significant effects were observed with TNF-alpha polymorphism alone; however, in combination with the IL-10 genotype, it increased the strength of these associations. Results show an association between the low IL-10 producer genotype and protection from RA; nevertheless, when other specific genetic and/or environmental factors trigger onset of RA, this genotype may predispose to development of anti-CCP+ RA disease with reduced response to prednisone treatment.


Inhibitory Receptor Expression on T Cells as a Marker of Disease Activity and Target to Regulate Effector Cellular Responses in Rheumatoid Arthritis.
Inhibitory receptors are essential for the regulation of effector immune responses and may play critical roles in autoimmune diseases. We evaluated whether inhibitory receptor expression on T cells from patients with rheumatoid arthritis (RA) were correlated with immune activation, disease activity, and response to treatment, as well as whether inhibitory receptor-mediated pathways were functional. Using flow cytometry, we performed extensive phenotypic and functional evaluation of CD4+ and CD8+ T cells from the blood and synovial fluid (SF) of RA patients ex vivo and after culture. The relationship of each parameter with the Disease Activity Score in 28 joints using the erythrocyte sedimentation rate (DAS28-ESR) and response to treatment was examined. In RA patients with low levels of T cell activation, inhibitory receptor expression showed an inverse relationship with the DAS28-ESR. The frequency of T cells expressing multiple inhibitory receptors was reduced in untreated RA patients but returned to normal levels in treated patients. RA patients who responded to treatment showed an augmented frequency of inhibitory receptor-expressing T cells that correlated with reduced inflammatory cytokine production in comparison to nonresponders. Higher frequencies of effector and memory T cells that expressed multiple inhibitory receptors were seen in SF than in peripheral blood. Notably, inhibitory pathways were operative in blood and synovial T cells from all RA patients, although cells from nonresponder patients were less sensitive to inhibition. Inhibitory receptor expression on T cells from RA patients is inversely correlated with effector T cell function and disease activity and may predict response to treatment. Furthermore, different inhibitory pathways are functional and cooperatively suppress synovial T cells, providing a rationale for new treatment strategies to regulate acute local inflammation.