Heliox with inhaled nitric oxide: a novel strategy for severe localized interstitial pulmonary emphysema in preterm neonatal ventilation.
We describe the combined use of inhaled nitric oxide and heliox (79% helium and 21% oxygen) as a rescue therapy for a critically ill infant with localized interstitial pulmonary emphysema and pulmonary hypertension. Conventional interventions were ineffective, not feasible, or unlikely to take effect in time, during this infant's acute critical illness. We added heliox based on its known pulmonary effects, and inhaled nitric oxide to improve oxygenation, after echocardiographic evidence of high right-ventricular pressure. The infant made a full recovery. To our knowledge this is the first case report of heliox and inhaled nitric oxide used simultaneously in localized interstitial pulmonary emphysema.


Effects of hypercapnia and flow-resistive loading on tracheal pressure during airway occlusion.
To determine whether the isometric force of concentration of the inspiratory muscles could be used to assess respiratory efferent neural activity, the tracheal pressure generated by the inspiratory muscles during airway occlusion (occluded tracheal pressure) was measured during progressive hypercapnia in anesthetized dogs breathing normally and breathing against added flow-resistive loads. Hypercapnia increased the peak end-inspiratory tracheal pressure and the occluded tracheal pressures generated 100, 200, and 300 ms after the onset of inspiration. The duration of the occluded inspiratory effort generally remained unchanged and the configuration of the pressure tracing was not affected. During normal breathing occluded tracheal pressures increased linearly with tidal volume and with the electrical activity of the diaphragm and the external intercostal muscles both before and after vagotomy. Inspiratory flow-resistive loading reduced the ventilatory response to CO2 but did not affect occluded tracheal pressures at any given PCO2 or the change in pressures with hypercapnia both before and after vagotomy. Similarly, expiratory flow-resistive loading failed to affect occluded tracheal pressures. These results suggest that occluded tracheal pressures measure respiratory efferent neural activity and can be used as indices of CO2 responsivity even during mechanical loading in anesthetized animals.


Predatory behaviour in synthetic protocell communities.
Recent progress in the chemical construction of colloidal objects comprising integrated biomimetic functions is paving the way towards rudimentary forms of artificial cell-like entities (protocells). Although several new types of protocells are currently available, the design of synthetic protocell communities and investigation of their collective behaviour has received little attention. Here we demonstrate an artificial form of predatory behaviour in a community of protease-containing coacervate microdroplets and protein-polymer microcapsules (proteinosomes) that interact via electrostatic binding. The coacervate microdroplets act as killer protocells for the obliteration of the target proteinosome population by protease-induced lysis of the protein-polymer membrane. As a consequence, the proteinosome payload (dextran, single-stranded DNA, platinum nanoparticles) is trafficked into the attached coacervate microdroplets, which are then released as functionally modified killer protocells capable of rekilling. Our results highlight opportunities for the development of interacting artificial protocell communities, and provide a strategy for inducing collective behaviour in soft matter microcompartmentalized systems and synthetic protocell consortia.


Nanographene charge trapping memory with a large memory window.
Nanographene is a promising alternative to metal nanoparticles or semiconductor nanocrystals for charge trapping memory. In general, a high density of nanographene is required in order to achieve high charge trapping capacity. Here, we demonstrate a strategy of fabrication for a high density of nanographene for charge trapping memory with a large memory window. The fabrication includes two steps: (1) direct growth of continuous nanographene film; and (2) isolation of the as-grown film into high-density nanographene by plasma etching. Compared with directly grown isolated nanographene islands, abundant defects and edges are formed in nanographene under argon or oxygen plasma etching, i.e. more isolated nanographene islands are obtained, which provides more charge trapping sites. As-fabricated nanographene charge trapping memory shows outstanding memory properties with a memory window as wide as ∼9 V at a relative low sweep voltage of ±8 V, program/erase speed of ∼1 ms and robust endurance of >1000 cycles. The high-density nanographene charge trapping memory provides an outstanding alternative for downscaling technology beyond the current flash memory.


Sliding-window analysis tracks fluctuations in amygdala functional connectivity associated with physiological arousal and vigilance during fear conditioning.
We evaluated whether sliding-window analysis can reveal functionally relevant brain network dynamics during a well-established fear conditioning paradigm. To this end, we tested if fMRI fluctuations in amygdala functional connectivity (FC) can be related to task-induced changes in physiological arousal and vigilance, as reflected in the skin conductance level (SCL). Thirty-two healthy individuals participated in the study. For the sliding-window analysis we used windows that were shifted by one volume at a time. Amygdala FC was calculated for each of these windows. Simultaneously acquired SCL time series were averaged over time frames that corresponded to the sliding-window FC analysis, which were subsequently regressed against the whole-brain seed-based amygdala sliding-window FC using the GLM. Surrogate time series were generated to test whether connectivity dynamics could have occurred by chance. In addition, results were contrasted against static amygdala FC and sliding-window FC of the primary visual cortex, which was chosen as a control seed, while a physio-physiological interaction (PPI) was performed as cross-validation. During periods of increased SCL, the left amygdala became more strongly coupled with the bilateral insula and anterior cingulate cortex, core areas of the salience network. The sliding-window analysis yielded a connectivity pattern that was unlikely to have occurred by chance, was spatially distinct from static amygdala FC and from sliding-window FC of the primary visual cortex, but was highly comparable to that of the PPI analysis. We conclude that sliding-window analysis can reveal functionally relevant fluctuations in connectivity in the context of an externally cued task.


[Vascular pathology in the aspect of sudden death in young adults and connective tissue dysplasia: anatomical, physiological, and morphological parallels].
Vascular pathology in young adults has during the last 10 years been diagnosed more often during forensic medical examination of sudden death. Major morphological alterations are revealed in cerebral vessels, coronary vessels, and at the level of the ascending portion of the aorta. Generally, in the young age there is no stenosing atherosclerosis inducing vascular lesions and the development of complications. It was determined that connective tissue dysplasia is pathology wherein weakness of the vascular wall is genetically preconditioned, thus promoting formation of vascular aneurysms and rupture of the latter under conditions of provoking factors such as going in for sports, physical loads, and psychoemotional stress.


Human V gamma 2V delta 2 T cells augment migration-inhibitory factor secretion and counteract the inhibitory effect of glucocorticoids on IL-1 beta and TNF-alpha production.
In immune cells, proinflammatory cytokine gene expression is regulated by glucocorticoids, whereas migration-inhibitory factor (MIF), a pleiotropic cytokine, has the unique property of counteracting the inhibitory effect of glucocorticoids on TNF-alpha and IL-1beta secretion. A few lines of evidence suggest that gammadelta T cells play an important role in immunoregulation. However, it is unknown whether human gammadelta T cells participate in regulating MIF secretion, and how gammadelta T cells, glucocorticoids, and cytokines converge to give a unified physiological response. In this study, we demonstrate that human Vgamma2Vdelta2 T cells augment MIF secretion. Remarkably, these Vgamma2Vdelta2 T cells, functioning similarly to MIF in part, counteracted inhibition of dexamethasone on production of IL-1beta and TNF-alpha. SCID mice reconstituted with human PBMC that were mock depleted of Vdelta2 T cells and repeatedly infected with lethal dose of Escherichia coli had shorter survival time than those reconstituted with PBMC that were depleted of Vdelta2 T cells. Thus, human Vgamma2Vdelta2 T cells are likely to play broad-spectrum roles in immunoregulation and immunopathology by influencing MIF secretion and the immunomodulatory function of glucocorticoids.


[Epidemiology of hepatitis A in Sardinia: prevalence of anti-HAV in a sample of junior and senior high school students].
In the developing countries, where socioeconomic and hygienic conditions are inadequate, hepatitis A is mainly a childhood infection. Improvements in environmental conditions cause a decline in the exposure to hepatitis A virus and a change in the age-distribution of the incidence. In the Sardinian population 3-11 years old a study conducted in 1987 showed that 3.8 percent of children were anti-HAV positive. In this research work we studied the anti-HAV prevalence in the population 13-20 years old. The study population consisted of all the children who attended the third year of the junior high school (13-16 years old) and the fifth year of the senior high school (17-20 years old) in 7 health districts of South-Sardinia. All schools were identified in each district. Our list consisted of 455 classes of the last year of secondary schools and of 242 classes of the last year of high schools. Our sample consisted of 20 classes of the last year of junior high schools and of 29 classes of the last year of senior high schools. We used a precoded questionnaire recording age, sex, number of households and father's years of schooling. Sera from 794 students were collected between May and June 1988 and stored at--20 degrees C. A ELISA test (ABBOTT) was used for detection of anti-HAV. The analysis has been carried out in accordance with the manufacturer's instructions. We determined the prevalence of anti-HAV according to age, sex and socioeconomic conditions. We calculated odds' ratios and their 95% confidence limits. We considered as not exposed the group in the most favourable level of exposure (father's highest educational level, lowest number of households). There is a sharp increase from an anti-HAV prevalence of 16.5 percent in students 13-16 years of age to 24.8 percent in students 17-20 years of age. As far as the persons 13-16 years of age are concerned we can recognize a significant difference between females (21.8 percent) and males (10.3 percent). In the other group of age, on the contrary, the difference is not significant.(ABSTRACT TRUNCATED AT 250 WORDS)


Generation of Ag2O micro-/nanostructures by pulsed excimer laser ablation of Ag in aqueous solutions of polysorbate 80.
A new route to synthesis of Ag(2)O micro-/nanostructures, including a mixture of cubes, pyramids, triangular plates, pentagonal rods, and bars, has been developed by pulsed excimer laser ablation of bulk silver in water using polysorbate 80 as surfactant. The polysorbate 80 played an important role in the formation of the Ag(2)O structures, and similar structures could be obtained in polysorbates 20 and 40 aqueous solutions. We have proposed a mechanism to explain the formation of Ag(2)O structures. This laser ablation method provides a unique approach to discover and fabricate new Ag(2)O morphologies.


Pathogenesis and prevalence of atherosclerosis in hypertensive patients.
Atherosclerosis is important because of its complications that include, for example, myocardial infarction, angina pectoris, stroke, aortic aneurysm, and intermittent claudication. Prospective studies have indicated that the risk-factor patterns for these complications are different in various parts of the cardiovascular tree. For stroke, hypertension is quantitatively the most important risk factor whereas, for intermittent claudication, smoking appears to be by far the most important risk factor. Myocardial infarction, however, is a complication for which a large number of risk factors emerge and none appears to be more important, at least from the statistical point of view, than the others. The prevalence of the different types of complications of atherosclerosis varies considerably among countries. Despite a high prevalence of stroke in Japan and a prevalence of hypertension equal to that in the United States, the prevalence of myocardial infarction is low. This indicates that the importance of hypertension as a risk factor for myocardial infarction is dependent on the presence of other risk factors, such as hypercholesterolemia. In the United States and several European countries, for instance Sweden, there has been a dramatic reduction in the yearly incidence of stroke since the beginning of the 1950s. The time sequence for this change and the change in the number of patients treated for hypertension have been similar in both the United States and Sweden. In contrast, the decrease in the incidence of myocardial infarction began about 15 years later in Sweden than in the United States. In Sweden, the reduced incidence of stroke can be explained by the reduction in the incidence of hemorrhagic stroke only.(ABSTRACT TRUNCATED AT 250 WORDS)


Passive range finder based on moiré images of Ronchi-like gratings with bilateral asymmetry.
A passive range-finding technique utilizing moiré images of Ronchi-like gratings with bilateral asymmetry, specifically fork gratings, is explored. Fork gratings are used as opposed to Ronchi gratings due to their asymmetry in the spatial and frequency domains, which enable the resolution of certain ambiguities in analysis that would otherwise be present. The two patterns which are convolved to produce the moiré image are the fork grating and the nailbed pattern of the pixelated detector. A fork grating of known size and spatial frequency is imaged with a detector of a fixed pixel pitch. Multiple analysis techniques are applied in frequency space, which lead to both relative and absolute measures of distance.


Correlation of Minimum Apparent Diffusion Coefficient and Maximum Standardized Uptake Value of the Primary Tumor with Clinicopathologic Characteristics in Endometrial Cancer.
To explore the correlation of the primary tumor's maximum standardized uptake value (SUVmax) and minimum apparent diffusion coefficient (ADCmin) with clinicopathologic features, and to determine their predictive power in endometrial cancer (EC). A total of 45 patients who had undergone staging surgery after a preoperative evaluation with 18F-fluorodeoxyglucose (FDG) positron emission tomography/computerized tomography (PET/CT) and diffusion-weighted magnetic resonance imaging (DW-MRI) were included in a prospective case-series study with planned data collection. Multiple linear regression analysis was used to determine the correlations between the study variables. The mean ADCmin and SUVmax values were determined as 0.72±0.22 and 16.54±8.73, respectively. A univariate analysis identified age, myometrial invasion (MI) and lymphovascular space involvement (LVSI) as the potential factors associated with ADCmin while it identified age, stage, tumor size, MI, LVSI and number of metastatic lymph nodes as the potential variables correlated to SUVmax. In multivariate analysis, on the other hand, MI was the only significant variable that correlated with ADCmin (p=0.007) and SUVmax (p=0.024). Deep MI was best predicted by an ADCmin cutoff value of ≤0.77 [93.7% sensitivity, 48.2% specificity, and 93.0% negative predictive value (NPV)] and SUVmax cutoff value of >20.5 (62.5% sensitivity, 86.2% specificity, and 81.0% NPV); however, the two diagnostic tests were not significantly different (p=0.266). Among clinicopathologic features, only MI was independently correlated with SUVmax and ADCmin. However, the routine use of 18F-FDG PET/CT or DW-MRI cannot be recommended at the moment due to less than ideal predictive performances of both parameters.


Physiological responses of great sturgeon (Huso huso) to different concentrations of 2-phenoxyethanol as an anesthetic.
The objective of the study was to evaluate the efficacy of 2-phenoxyethanol (2-PE) as an anesthetic in great sturgeon under two experiments. First, fish were exposed to 0.1, 0.3, 0.5, 0.7 and 0.9 mL/L 2-PE, and time to induction (deep anesthesia) and recovery from anesthesia were measured. At concentration of 0.1 mL/L, 2-PE failed to induce deep anesthesia in fish, whereas at concentrations of 0.7 and 0.9 mL/L, all the fish were anaesthetized within 3 min of exposure. For assessing the impact of effective concentrations of 2-PE on physiological responses of great sturgeon, hematological indices, plasma metabolites, electrolytes, enzymes and cortisol levels were measured. The use of 2-PE induces a significant increase in RBC values at 0.3 mL/L concentration and a parallel increase in hemoglobin and hematocrit values. 2-PE anesthesia had no effect on WBC, MCV, MCH and MCHC levels when compared to control group. Serum glucose, cholesterol and cortisol levels were significantly high in 0.3 and 0.5 mL/L 2-PE. Moreover, AST levels were increased in fish exposed to the 0.3 mL/L 2-PE comparing with the control group (P < 0.05). There were no significant differences in serum levels of total protein, triglycerides, ALP, ALT, Cl(-), Na(+), K(+), Ca(2+) and Mg(2+). In this study, alteration in hematological and serum biochemical indices was time-dependent. This study demonstrates that rapid induction of deep anesthesia with a relatively high concentration of 2-PE (0.7 and 0.9 mL/L) was associated with the lowest effects on the hematological and serum biochemical indices in great sturgeon and therefore would be recommended as eligible doses for hematological studies in this species.


Targeted deletion of Capn4 in cells of the chondrocyte lineage impairs chondrocyte proliferation and differentiation.
Calpains are calcium-dependent intracellular cysteine proteases, which include ubiquitously expressed mu- and m-calpains. Both calpains are heterodimers consisting of a large catalytic subunit and a small regulatory subunit. The calpain small subunit encoded by the gene Capn4 directly binds to the intracellular C-terminal tail of the receptor for the parathyroid hormone (PTH) and PTH-related peptide and modulates cellular functions in cells of the osteoblast lineage in vitro and in vivo. To investigate a physiological role of the calpain small subunit in cells of the chondrocyte lineage, we generated chondrocyte-specific Capn4 knockout mice. Mutant embryos had reduced chondrocyte proliferation and differentiation in embryonic growth plates compared with control littermates. In vitro analysis further revealed that deletion of Capn4 in cells of the chondrocyte lineage correlated with impaired cell cycle progression at the G(1)/S transition, reduced cyclin D gene transcription, and accumulated cell cycle proteins known as calpain substrates. Moreover, silencing of p27(Kip1) rescued an impaired cell growth phenotype in Capn4 knockdown cells, and reintroducing the calpain small subunit partially normalized cell growth and accumulated cyclin D protein levels in a dose-dependent manner. Collectively, our findings suggest that the calpain small subunit is essential for proper chondrocyte functions in embryonic growth plates.


Quantitative monitoring of capsular contraction around smooth and textured implants.
The purpose of this study was to assess the efficacy of textured silicone implants in reducing the incidence of capsular contracture. Each of 10 female New Zealand, albino rabbits received 2 saline-filled implants, 1 on either side of the lateral chest wall. The surface of 1 implant was smooth silicone, whereas the other implant's surface was textured silicone. Magnetic resonance imaging (MRI) scans of the implants were performed at 0, 9, 17, 26, 34, and 40 weeks after implantation. Data from the MRI scans were used to calculate the effective surface area of implants at each analysis interval. This technique provided a noninvasive method of monitoring implant contraction as a function of time. Eight rabbits completed the study. Four of 8 smooth implants developed contractures, whereas none of the textured implants developed contracture. For the 4 smooth implants that developed contractures, MRI scans calculated 72 +/- 12% contraction at 17 weeks, but the Baker palpation test detected only mild firmness. From 17 to 40 weeks, the mean percentage of contraction for these implants changed minimally, but their mean Baker score increased from mild to severe (II to IV). Quantitative data from MRI scans were much more predictive of final implant contraction than palpation (Baker test), applanation tonometry, or indentation tonometry. The latter two tests only detected the final stages of severe implant contraction.


One-step repair of a large congenital upper eyelid coloboma using a bilamellar graft in an infant: a case report and literature review.
Large congenital lid colobomas are traditionally repaired using 1- or 2-step vascularized flap-graft combinations. However, visual axis occlusion for weeks is a severe problem in small children and recent reports suggest that the flap pedicle does not contribute to blood perfusion. A "one-step" substitute for large lid defects has recently been reported in animals and humans, demonstrating the viability of a bilamellar autograft alone. We present an alternative "one-step" reconstructive approach in a 6-month-old infant who had a centrally-located large upper eyelid defect resulting from a congenital coloboma. The free full-thickness bilamellar autograft was harvested from the contralateral upper eyelid. The follow-up time was 48 months. Cosmetic and functional results were good, the bilamellar graft survived, and there was no graft ischemia, necrosis, or rejection. The boy developed madarosis, lid notching, and mild contour irregularity but needed no reoperation since the parent was satisfied with the surgical result. A free bilamellar eyelid autograft seems to be an outstanding alternative to both "conventional 2-step" and "modern 1-step" options for the reconstruction of large colobomatous eyelid openings, especially in young infants who cannot tolerate visual axis blockage. It is an easy, practical, fast, and effective technique that also saves cost in health care.


Effect of beryllium chloride on porphyrin metabolism in pregnant mice administered by subcutaneous injection.
The effect of beryllium (Be) compounds on porphyrins was investigated in pregnant mice. The blood protoporphyrin (Proto) and zinc protoporphyrin (Zn Proto) concentrations were increased in pregnancy. Regardless of pregnancy or nonpregnancy, the Proto concentration was decreased after Be injection. Delta-aminolevulinic acid dehydratase (ALA-D) and porphobilinogen deaminase (PBG-D) activities in blood were significantly elevated in the pregnant untreated (Con-pregnant) group, compared to the nonpregnant mice untreated (Con-nonpregnant) and nonpregnant mice treated with Be (Be-nonpregnant) groups. The blood ALA-D activity of the pregnant mice treated with Be (Be-pregnant group) tended to decrease, compared to Con-pregnant group. The blood PBG-D activity in the Be-pregnant group was significantly lower compared with that of the Con-pregnant group. The ALA-D and PBG-D activities in the spleen were also significantly elevated in the Con-pregnant group, compared to nonpregnant groups. However, it was noted that these values in the Be-pregnant group were almost the same as that of the Con-nonpregnant group and were significantly lower than that in the Con-pregnant group. The elevation of ALA-D and PBG-D activities in the blood and spleen, which play a role in the hematopoietic function of mice, was observed in the Con-pregnant mice compared to the nonpregnant mice. However, the phenomenon was not observed in the Be-pregnant mice, it suggesting that Be suppressed the pregnancy-induced increase in hematopoietic function.


Sparse Representation in Awake Auditory Cortex: Cell-type Dependence, Synaptic Mechanisms, Developmental Emergence, and Modulation.
Sparse representation is considered an important coding strategy for cortical processing in various sensory modalities. It remains unclear how cortical sparseness arises and is being regulated. Here, unbiased recordings from primary auditory cortex of awake adult mice revealed salient sparseness in layer (L)2/3, with a majority of excitatory neurons exhibiting no increased spiking in response to each of sound types tested. Sparse representation was not observed in parvalbumin (PV) inhibitory neurons. The nonresponding neurons did receive auditory-evoked synaptic inputs, marked by weaker excitation and lower excitation/inhibition (E/I) ratios than responding cells. Sparse representation arises during development in an experience-dependent manner, accompanied by differential changes of excitatory input strength and a transition from unimodal to bimodal distribution of E/I ratios. Sparseness level could be reduced by suppressing PV or L1 inhibitory neurons. Thus, sparse representation may be dynamically regulated via modulating E/I balance, optimizing cortical representation of the external sensory world.


Cervical spondylosis: most common cause of position and vibratory sense loss.
At present, spondylosis of the cervical spine is the most common cause of loss of position and vibratory sense. The loss is asymmetric in regard to location and degree of involvement and results from intermittent contusion of the posterior columns against the lamina in persons having congenital narrow spinal canal. Subluxation of the vertebrae, reversal of cervical lordosis, and a hypertrophic ligamentum flavum further compromise the canal. Measurement of the anteroposterior diameter of the spinal canal is meaningful, as a narrow canal correlates best with the clinical symptoms. Not all patients with a narrow sagittai diameter have clinical signs, but all patients with clinical signs have a narrow sagittal diameter. Immobilization of the cervical spine in partial flexion with a light collar is the proper treatment in the absence of spinal fluid block.


A comparison of artificial saliva and pilocarpine in radiation-induced xerostomia.
Twenty patients with radiation-induced xerostomia were entered into a prospective randomized crossover study comparing a mucin-based artificial saliva (Saliva Orthana) and a mouthwash containing pilocarpine (5 mg three times a day). Overall the patients found that the pilocarpine mouthwash was more effective than the artificial saliva in relieving their symptoms (p = 0.04), and 47 per cent of the patients wanted to continue with this treatment after the study had finished.


Event-Based Variance-Constrained ${\mathcal {H}}_{\infty }$ Filtering for Stochastic Parameter Systems Over Sensor Networks With Successive Missing Measurements.
This paper is concerned with the distributed filtering problem for a class of discrete time-varying stochastic parameter systems with error variance constraints over a sensor network where the sensor outputs are subject to successive missing measurements. The phenomenon of the successive missing measurements for each sensor is modeled via a sequence of mutually independent random variables obeying the Bernoulli binary distribution law. To reduce the frequency of unnecessary data transmission and alleviate the communication burden, an event-triggered mechanism is introduced for the sensor node such that only some vitally important data is transmitted to its neighboring sensors when specific events occur. The objective of the problem addressed is to design a time-varying filter such that both the requirements and the variance constraints are guaranteed over a given finite-horizon against the random parameter matrices, successive missing measurements, and stochastic noises. By recurring to stochastic analysis techniques, sufficient conditions are established to ensure the existence of the time-varying filters whose gain matrices are then explicitly characterized in term of the solutions to a series of recursive matrix inequalities. A numerical simulation example is provided to illustrate the effectiveness of the developed event-triggered distributed filter design strategy.


TLR2 agonists and their structure-activity relationships.
Agonists of Toll-like Receptor 2 (TLR2) are attractive synthetic targets due to their use as adjuvants in immunotherapies to treat various diseases notably, cancer. An indepth understanding of TLR2 agonist structure-activity relationships is therefore advantageous for the methodical design of vaccines targetting the TLR2 machinery. This review aims to collate and discuss the literature regarding synthetic studies towards TLR2 agonists and the structure-activity relationships thereof. It is hoped that interested readers will gain a holistic understanding of this topic, and will prompt further efforts towards finding effective agonists of TLR2.


Membrane proteins: always an insoluble problem?
Membrane proteins play crucial roles in cellular processes and are often important pharmacological drug targets. The hydrophobic properties of these proteins make full structural and functional characterization challenging because of the need to use detergents or other solubilizing agents when extracting them from their native lipid membranes. To aid membrane protein research, new methodologies are required to allow these proteins to be expressed and purified cheaply, easily, in high yield and to provide water soluble proteins for subsequent study. This mini review focuses on the relatively new area of water soluble membrane proteins and in particular two innovative approaches: the redesign of membrane proteins to yield water soluble variants and how adding solubilizing fusion proteins can help to overcome these challenges. This review also looks at naturally occurring membrane proteins, which are able to exist as stable, functional, water soluble assemblies with no alteration to their native sequence.


[The growth behavior of mouse fibroblasts on intraocular lens surface of various silicone and PMMA materials].
Experience with intraocular lenses (IOL) made of PMMA dates back ca. 40 years, while silicone IOLs have been in use for only about 10 years. The biocompatibility of PMMA and silicone caoutchouc was tested in a comparative study investigating the growth of mouse fibroblasts on different IOL materials. Spectrophotometric determination of protein synthesis and liquid scintillation counting of DNA synthesis were carried out. The spreading of cells was planimetrically determined, and the DNA synthesis of individual cells in direct contact with the test sample was tested. The results showed that the biocompatibility of silicone lenses made of purified caoutchouc is comparable with that of PMMA lenses; there is no statistically significant difference. However, impurities arising during material synthesis result in a statistically significant inhibition of cell growth on the IOL surfaces.


Pilot study for cystic fibrosis neonatal screening: the Cuban experience.
Background In Cuba, no screening program for cystic fibrosis (CF) has been implemented yet. The ultramicro enzyme-linked immunosorbent assay (UMELISA)® TIR NEONATAL has been developed for the measurement of immunoreactive trypsin (IRT) in dried blood spots on filter paper. The analytical performance of the kit was evaluated in the national network of laboratories. Methods Newborn dried blood samples (DBS) were evaluated in 16 laboratories. An IRT/IRT/DNA protocol was followed using a cut-off value of 50 ng/mL. The mean, median and percentiles of the distribution were calculated and a two-sample t-test with unequal variance was used for statistical analysis. Influence of perinatal factors on IRT levels was analyzed. Results From January to June 2018, 6470 newborns were studied, obtaining a mean IRT value of 12.09 ng/mL (ranging 0-358 ng/mL) and a median of 8.99 ng/mL. Fifty-two samples (0.78%) were above the cut-off level and 16 samples (0.24%) were elevated in the re-screening process. One of them was confirmed positive by molecular biology (phe508del/c.3120 + 1G > A), constituting the first newborn screened and diagnosed early in Cuba. Second DBS samples were collected on average at 14 days and processed in the laboratory at 16 days of birth. Significant differences were observed (p < 0.05) when evaluating the influence of gender, birth weight (BW) and gestational age (GA) on the IRT values. Lower IRT concentrations were found in samples processed after 10 days of collection. Conclusions The performance of UMELISA® TIR NEONATAL in the laboratories has been satisfactory; hence CF newborn screening (NBS) was extended throughout the country from January 2019.


Nonaqueous magnetic nanoparticle suspensions with controlled particle size and nuclear magnetic resonance properties.
We report the preparation of monodisperse maghemite (gamma-Fe2O3) nanoparticle suspensions in heptane, by thermal decomposition of iron(III) acetylacetonate in the presence of oleic acid and oleylamine surfactants. By varying the surfactant/Fe precursor mole ratio during synthesis, control was exerted both over the nanocrystal core size, in the range from 3 to 6 nm, and over the magnetic properties of the resulting nanoparticle dispersions. We report field-cycling 1H NMR relaxation analysis of the superparamagnetic relaxation rate enhancement of nonaqueous suspensions for the first time. This approach permits measurement of the relaxivity and provides information on the saturation magnetization and magnetic anisotropy energy of the suspended particles. The saturation magnetization was found to be in the expected range for maghemite particles of this size. The anisotropy energy was found to increase significantly with decreasing particle size, which we attribute to increased shape anisotropy. This study can be used as a guide for the synthesis of maghemite nanoparticles with selected magnetic properties for a given application.


Ripeness Prediction of Postharvest Kiwifruit Using a MOS E-Nose Combined with Chemometrics.
Postharvest kiwifruit continues to ripen for a period until it reaches the optimal "eating ripe" stage. Without damaging the fruit, it is very difficult to identify the ripeness of postharvest kiwifruit by conventional means. In this study, an electronic nose (E-nose) with 10 metal oxide semiconductor (MOS) gas sensors was used to predict the ripeness of postharvest kiwifruit. Three different feature extraction methods (the max/min values, the difference values and the 70th s values) were employed to discriminate kiwifruit at different ripening times by linear discriminant analysis (LDA), and results showed that the 70th s values method had the best performance in discriminating kiwifruit at different ripening stages, obtaining a 100% original accuracy rate and a 99.4% cross-validation accuracy rate. Partial least squares regression (PLSR), support vector machine (SVM) and random forest (RF) were employed to build prediction models for overall ripeness, soluble solids content (SSC) and firmness. The regression results showed that the RF algorithm had the best performance in predicting the ripeness indexes of postharvest kiwifruit compared with PLSR and SVM, which illustrated that the E-nose data had high correlations with overall ripeness (training: R² = 0.9928; testing: R² = 0.9928), SSC (training: R² = 0.9749; testing: R² = 0.9143) and firmness (training: R² = 0.9814; testing: R² = 0.9290). This study demonstrated that E-nose could be a comprehensive approach to predict the ripeness of postharvest kiwifruit through aroma volatiles.


Effect of solvent composition on transformation of micelles to vesicles of rod-coil poly(n-hexyl isocyanate-block-2-vinylpyridine) diblock copolymers.
The self-aggregation behavior of an amphiphilic rod-coil block copolymer of poly(n-hexyl isocyanate-block-2-vinylpyridine) (PHIC(189)-b-P2VP(228)) (f(P2VP) = 0.78, M(n) = 24.5K) in a tetrahydrofuran (THF)/water system was examined using dynamic light scattering (DLS), transmission electron microscopy (TEM), and field emission scanning electron microscopy (FE-SEM). The presence of a certain amount of water in the THF-based polymer solution induced a morphological transition from spherical solid micelles to open mouth platelike vesicles. The size of the aggregates increased with an increase in water content in the mixed solvent of THF/water. In the range of 30-40% water, the polymer formed vesicles with an interdigitated architecture of poly(n-hexyl isocyanate) (PHIC) at the center of the membrane and with the poly(2-vinylpyridine) (P2VP) block forming the outer layers and pointing toward the solvent. However, at higher water contents, the thickness of the bilayer increased due to the rearrangement of the vesicle membrane from a flip-flop to a lamellar architecture. After the degradation of the PHIC from the vesicles at basic pH, hollow spherical aggregates remained stable. After removing the THF from the mixed solvent using dialysis, large-sized compound vesicles were formed.


Combined Blockade of T Cell Immunoglobulin and Mucin Domain 3 and Carcinoembryonic Antigen-Related Cell Adhesion Molecule 1 Results in Durable Therapeutic Efficacy in Mice with Intracranial Gliomas.
BACKGROUND Glioblastoma multiforme (GBM) evades immune surveillance by inducing immunosuppression via receptor-ligand interactions between immune checkpoint molecules. T cell immunoglobulin and mucin domain 3 (Tim-3) is a key checkpoint receptor responsible for exhaustion and dysfunction of T cells and plays a critical role in immunosuppression. Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) has been recently identified as a heterophilic ligand for Tim-3. MATERIAL AND METHODS We established an intracranial GBM model using C57BL/6 mice and GL261 cells, and treated the mice with single or combined monoclonal antibodies (mAbs) against Tim-3/CEACAM1. The CD4+, CD8+, and regulatory T cells in brain-infiltrating lymphocytes were analyzed using flow cytometry, and the effector function of T cells was assessed using ELISA. We performed a rechallenge by subcutaneous injection of GL261 cells in the "cured" (>90 days post-orthotopic tumor implantation) and naïve mice. RESULTS The mean survival time in the control, anti-Tim-3, anti-CEACAM1, and combined treatment groups was 29.8, 43.4, 42.3, and 86.0 days, respectively, with 80% of the mice in the combined group becoming long-term survivors showing immune memory against glioma cells. Infiltrating CD4+ and CD8+ T cells increased and immunosuppressive Tregs decreased with the combined therapy, which resulted in a markedly elevated ratio of CD4+ and CD8+ cells to Tregs. Additionally, plasma IFN-γ and TGF-β levels were upregulated and downregulated, respectively. CONCLUSIONS Our data indicate that combined blockade of Tim-3 and CEACAM1 generates robust therapeutic efficacy in mice with intracranial tumors, and provides a promising option for GBM immunotherapy.


LXR-dependent regulation of macrophage-specific reverse cholesterol transport is impaired in a model of genetic diabesity.
Diabesity and fatty liver have been associated with low levels of high-density lipoprotein cholesterol, and thus could impair macrophage-specific reverse cholesterol transport (m-RCT). Liver X receptor (LXR) plays a critical role in m-RCT. Abcg5/g8 sterol transporters, which are involved in cholesterol trafficking into bile, as well as other LXR targets, could be compromised in the livers of obese individuals. We aimed to determine m-RCT dynamics in a mouse model of diabesity, the db/db mice. These obese mice displayed a significant retention of macrophage-derived cholesterol in the liver and reduced fecal cholesterol elimination compared with nonobese mice. This was associated with a significant downregulation of the hepatic LXR targets, including Abcg5/g8. Pharmacologic induction of LXR promoted the delivery of total tracer output into feces in db/db mice, partly due to increased liver and small intestine Abcg5/Abcg8 gene expression. Notably, a favorable upregulation of the hepatic levels of ABCG5/G8 and NR1H3 was also observed postoperatively in morbidly obese patients, suggesting a similar LXR impairment in these patients. In conclusion, our data show that downregulation of the LXR axis impairs cholesterol transfer from macrophages to feces in db/db mice, whereas the induction of the LXR axis partly restores impaired m-RCT by elevating the liver and small intestine expressions of Abcg5/g8.


Exome sequencing identifies compound heterozygous mutations in CYP4V2 in a pedigree with retinitis pigmentosa.
Retinitis pigmentosa (RP) is a heterogeneous group of progressive retinal degenerations characterized by pigmentation and atrophy in the mid-periphery of the retina. Twenty two subjects from a four-generation Chinese family with RP and thin cornea, congenital cataract and high myopia is reported in this study. All family members underwent complete ophthalmologic examinations. Patients of the family presented with bone spicule-shaped pigment deposits in retina, retinal vascular attenuation, retinal and choroidal dystrophy, as well as punctate opacity of the lens, reduced cornea thickness and high myopia. Peripheral venous blood was obtained from all patients and their family members for genetic analysis. After mutation analysis in a few known RP candidate genes, exome sequencing was used to analyze the exomes of 3 patients III2, III4, III6 and the unaffected mother II2. A total of 34,693 variations shared by 3 patients were subjected to several filtering steps against existing variation databases. Identified variations were verified in the rest family members by PCR and Sanger sequencing. Compound heterozygous c.802-8_810del17insGC and c.1091-2A>G mutations of the CYP4V2 gene, known as genetic defects for Bietti crystalline corneoretinal dystrophy, were identified as causative mutations for RP of this family.


Prostate Cancer, Version 4.2023, NCCN Clinical Practice Guidelines in Oncology.
The NCCN Guidelines for Prostate Cancer provide a framework on which to base decisions regarding the workup of patients with prostate cancer, risk stratification and management of localized disease, post-treatment monitoring, and treatment of recurrence and advanced disease. The Guidelines sections included in this article focus on the management of metastatic castration-sensitive disease, nonmetastatic castration-resistant prostate cancer (CRPC), and metastatic CRPC (mCRPC). Androgen deprivation therapy (ADT) with treatment intensification is strongly recommended for patients with metastatic castration-sensitive prostate cancer. For patients with nonmetastatic CRPC, ADT is continued with or without the addition of certain secondary hormone therapies depending on prostate-specific antigen doubling time. In the mCRPC setting, ADT is continued with the sequential addition of certain secondary hormone therapies, chemotherapies, immunotherapies, radiopharmaceuticals, and/or targeted therapies. The NCCN Prostate Cancer Panel emphasizes a shared decision-making approach in all disease settings based on patient preferences, prior treatment exposures, the presence or absence of visceral disease, symptoms, and potential side effects.


Bis(naphthobipyrrolyl)methene-derived hexapyrrolic BODIPY as a single-molecule helicate with near-infrared emission.
Helically twisted bis(naphthobipyrrolyl)methene-derived open-chain hexapyrroles have been synthesized as HCl salts and the corresponding BODIPY. Their solid-state structures elucidated by single-crystal X-ray diffraction analysis clearly showed the presence of intramolecular hydrogen bonds, which were concluded to play a pivotal role in stabilizing the twisted conformation. Both molecules were observed to be NIR active, with the BODIPY moiety emission extending beyond 800 nm.


Effect of heparin on infection of cells by porcine reproductive and respiratory syndrome virus.
To investigate whether heparin has any effect on the growth of porcine reproductive and respiratory syndrome virus (PRRSV). 2 isolates of PRRSV, and as control viruses, 1 isolate of pseudorabies virus (PRV) and 1 isolate of parainfluenza 3 virus (PIV-3). Plaque assays, using a continuous cell line (MARC-145) derived from African green monkey kidney cell line (MA104), were performed for determination of inhibitory effect of heparin on PRRSV, PRV, and PIV-3. The effect of various doses of heparin and heparinase on the growth of PRRSV, PRV, and PIV-3 was evaluated and compared. In each experiment, value were expressed as the mean value for duplicate samples. The number of plaques formed by PRRSV and PRV was reduced to 24 to 25 and 15% of the untreated control (100%), respectively, by 1 U of heparin/ml, but could not be reduced below 6 to 7 and 3%, respectively, by use of concentrations up to 50 U/ml. An inhibitory effect of heparin, at a concentration up to 50 U/ml, was not observed on PIV-3. Delaying addition of heparin for 30 minutes after the addition of PRRSV and PRV reduced plaque formation by 48 to 51 and 68%, respectively, compared with 91 to 92 and 95%, respectively, if heparin was added at the time of infection. In addition, most PRRSV added was retained by heparin beads, as was PRV. Heparinase treatment of MARC-145 cells reduced the number of PRRSV-, as well as PRV-induced plaques. On the other hand, the number of PIV-3-induced plaques did not decrease after treatment of MARC-145 cells with heparinase. Addition of heparin to PRRSV or to the MARC-145 cells before virus inoculation and treatment of the cells with heparinase prevented the virus from infecting the cells.


Evaluation of 60Co Irradiation on Volatile Components of Turmeric (Curcumae Longae Rhizoma) Volatile Oil with GC-IMS.
60Co irradiation is an efficient and rapid sterilization method. The aim of this work is to determine the changes in essential-oil composition under different irradiation intensities of 60Co and to select an appropriate irradiation dose with GC-IMS. Dosages of 0, 5, and 10 kGy of 60Co were used to analyze turmeric (Curcumae Longae Rhizoma) volatile oil after 60Co irradiation (named JH-1, JH-2, and JH-3). The odor fingerprints of volatile organic compounds in different turmeric volatile oil samples were constructed by headspace solid-phase microextraction and gas chromatography-ion mobility spectrometry (GC-IMS) after irradiation. The differences in odor fingerprints of volatile organic compounds (VOCs) were compared by principal component analysis (PCA). The results showed that 97 volatile components were detected in the volatile oil of Curcuma longa, and 64 components were identified by database retrieval. With the change in irradiation intensity, the volatile compounds in the three turmeric volatile oil samples were similar, but the peak intensity was significantly different, which was attributed to the change in compound composition and content caused by different irradiation doses. In addition, the principal component analysis showed that JH-2 and JH-3 were relatively correlated, while JH-1 and JH-3 were far from each other. In general, different doses of 60Co irradiation can affect the content of volatile substances in turmeric volatile oil. With the increase in irradiation dose, the peak area decreased, and so the irradiation dose of 5 kGy/min was better. It is shown that irradiation technology has good application prospects in the sterilization of foods with volatile components. However, we must pay attention to the changes in radiation dose and chemical composition.


Socioeconomic Gradient in Childhood Obesity and Hypertension: A Multilevel Population-Based Study in a Chinese Community.
This study aims to assess evidence for any socioeconomic gradients in childhood obesity and hypertension in a population-representative sample in Hong Kong, China. The data of a stratified random sampled growth survey collected in 2005-2006 was matched with a population by-census. Obesity was defined using the International Obesity Task Force standard and hypertension was defined using the Hong Kong norm table. Family socioeconomic status (SES) was measured by maternal education level. Neighbourhood SES was measured by median household income of the neighbourhood. Multilevel Poisson regression models with robust standard error were used to test the association. Body mass indices of children's parents were included as potential confounders. Intra-school/neighbourhood correlations were adjusted using random factors. Totally 14842 children (age 6-19 years) included in the analysis, in which 16.6% of them were overweight or obese. Children whose mother only completed secondary school or below had higher risk of childhood obesity (RR 1.41, 95% CI 1.13-1.76, p = 0.003) and hypertension (RR 1.18, 95% CI 1.01-1.36, p = 0.03). Meanwhile, children in the lowest neighbourhood SES group had higher risk of childhood underweight (RR 1.61, 95% CI 1.04-2.49, p = 0.03), overweight (RR 1.35, 95% CI 1.05-1.72, p = 0.02), and obesity (RR 2.07, 95% CI 1.11-3.88, p = 0.02). Socioeconomic gradient in childhood obesity and hypertension existed in Hong Kong, one of the most developed cities in China. These results have implications for policymakers and public health experts and highlight the need to monitor trends in other parts of China.


Evaluation of apically extruded debris in conventional and rotary instrumentation techniques.
The purpose of this in vitro study was to compare two common techniques, conventional and rotary instrumentation, in debris extrusion. Two hundred mature human teeth with mature apices and less than 15 degrees of root curvature as determined by Schneider's method were selected for use. Teeth were randomly divided into four groups of 50 teeth each and prepared using step-back instrumentation and profile 4 percent Taper Series 29 rotary system at three different speeds: 1,000 rpm, 8,000 rpm and 24,000 rpm. All of the speeds were reduced by contra angle to 1/6 rpm. Extruded debris were collected on pre-weighed vials containing distilled water through the apical foramen during instrumentation. The vials were then dried in a microwave oven at 150 degrees Celsius and weighed again. The difference was recorded as "the weight of the extruded debris." Eight control vials, two for each group, were weighed and filled with distilled water, dried, and reweighed. The mean weight of extruded debris for each group was statistically analyzed. T-student, unilateral variance analysis and Pearson tests were used in this study. Step-back instrumentation (Group 1) produced significantly more debris than other groups. The difference in the amount of debris produced among rotary groups was not significant. Rotary technique could reduce the amount of debris extrusion.


Enabling pharmacogenomic clinical trials through sampling.
Discussion and output from the US FDA and the pharmaceutical industry from the Drug Information Association/FDA 5th Workshop in a series on pharmacogenomics entitled: 'Generating and Weighing Evidence in Drug Development and Regulatory Decision Making'. A major topic area at the 5th FDA/Industry Workshop on Pharmacogenomics, February 2-4, 2010 in Bethesda (MD, USA), was enabling pharmacogenomic clinical trials through collection of future use samples. The importance of the collection of samples with permission for future analyses was affirmed by both industry and the FDA. In addition, current barriers for the collection of such samples were detailed and possible solutions for overcoming barriers at sites, as well as globally within countries, were discussed. The importance of international concordance on collection of these samples was emphasized, and potential areas for industry to harmonize sample collection practices. A standalone workshop on issues related to sampling was determined to be a key step for solving issues related to future use sample collection during drug development.


Metastasis after intravenous inoculation of highly metastatic variants of mouse tumors and the effects of several antitumor drugs on the tumors.
Tumor metastasis was examined after iv inoculation of highly metastatic variants of mouse tumors. Highly metastatic variants, B16-F10 and B16-BL6, of B16 melanoma origin and colon 26 NL-17 of colon adenocarcinoma 26 origin were used in the experiments. Formation of pulmonary metastasis was influenced by the mouse strain and age. Based on the results, experimental metastasis systems of these tumors were established, and the effects of chemotherapeutic agents were examined. 5-Fluorouracil and adriamycin were significantly effective for the suppression of pulmonary metastasis. The degrees of suppression of metastasis by these drugs were different with different tumor variants, but the sensitivity of the metastasis to the drug was similar to that of the parent tumor. Several aspects of the chemotherapy of metastasis are discussed.


Tissue flow cytometry immunophenotyping in the diagnosis and classification of non-Hodgkin's lymphomas: a retrospective evaluation of 1,792 cases.
A retrospective analysis of 1,792 solid tissues suggestive of lymphoma, submitted over a 12-year period, was carried out and flow cytometry (FC) results were compared with histologic findings. The final histologic diagnosis of cases documented in this report is as follows: 1,270 non-Hodgkin's lymphomas (NHL); 17 composite lymphomas; four NHL plus carcinomas; five post-transplant lymphoproliferative disorders; 105 Hodgkin's lymphomas (HL); eight acute leukemias; 42 tissue cancers; and 341 non-neoplastic diseases. A strong correlation between morphology and FC data was observed among hematological malignancies (1,268/1,304, 97.2%) with the exception of HL. Among B-NHL, FC detection of clonally restricted B-cell allowed the identification of lymphomas that were not histologically clear and the differential diagnosis between follicular lymphoma and reactive hyperplasia. A high correlation level (r = 0.83; P < 0.0001) was obtained in comparing proliferation results obtained by FC and immunohistochemistry. Among T-NHL, FC detection of an aberrant phenotype direct histologic diagnosis in cases having less than 20% of neoplastic cells. In nine cases, FC suggested the need to evaluate a neoplastic population, not morphologically evident. Results show that FC routinely performed on tissue samples suspected of lymphomas is a fundamental adjunct to morphology in the diagnosis of NHL and may enhance the performance of the histologic evaluation so as to achieve the final diagnosis. To the best of our knowledge, this is the first report in the literature of a wide series of tissues also studied by FC.


A Randomized Controlled Trial on the Effects of a 12-Week High- vs. Low-Intensity Exercise Intervention on Hippocampal Structure and Function in Healthy, Young Adults.
Physical exercise affects hippocampal structure and function, but the underlying neural mechanisms and the effects of exercise intensity remain incompletely understood. Therefore, we undertook a comprehensive, multi-modal 3T and 7T MRI randomized controlled trial (Netherlands Trial Register - NL5847) in which we randomized 52 young, non-athletic volunteers to a 12-week low- or high-intensity exercise program. Using state-of-the-art methods, we investigated changes in hippocampal volume, as well as changes in vasculature, neuro-metabolites, and peripheral growth factors as potential underpinnings. Cardiorespiratory fitness improved over time (p < 0.001), but no interaction with exercise intensity was found (p = 0.48). Accordingly, we did not observe significant interactions between exercise condition and time on MRI measures (all p > 0.06). However, we found a significant decrease in right hippocampal volume (p < 0.01), an increase in left hippocampal glutathione (p < 0.01), and a decrease of left hippocampal cerebral blood volume (p = 0.01) over time, regardless of exercise condition. Additional exploratory analyses showed that changes in brain-derived neurotrophic factor (p = 0.01), insulin-like growth-factor (p = 0.03), and dorsal anterior cingulate cortex N-acetyl-aspartate levels (p = 0.01) were positively associated with cardiorespiratory fitness changes. Furthermore, a trend toward a positive association of fitness and gray-matter cerebral blood flow (p = 0.06) was found. Our results do not provide evidence for differential effects between high-intensity (aerobic) and low-intensity (toning) exercise on hippocampal structure and function in young adults. However, we show small but significant effects of exercise on hippocampal volume, neurometabolism and vasculature across exercise conditions. Moreover, our exploratory results suggest that exercise might not specifically only benefit hippocampal structure and function, but rather has a more widespread effect. These findings suggest that, in agreement with previous MRI studies demonstrating moderate to strong effects in elderly and diseased populations, but none to only mild effects in young healthy cohorts, the benefits of exercise on the studied brain measures may be age-dependent and restorative rather than stimulatory. Our study highlights the importance of a multi-modal, whole-brain approach to assess macroscopic and microscopic changes underlying exercise-induced brain changes, to better understand the role of exercise as a potential non-pharmacological intervention.


Effects of four taurine-conjugated bile acids on mucosal uptake and lymphatic absorption of cholesterol in the rat.
The importance of the bile acid structure on mucosal uptake and lymphatic absorption of cholesterol was investigated using four different taurine-conjugated bile acids. Pure synthetic conjugates of a trihydroxy bile acid, taurocholate, and three dihydroxy bile acids, tauroursodeoxycholate, taurochenodeoxycholate, and taurodeoxycholate were used to completely solubilize [14C]cholesterol and polar lipids for steady rate intraduodenal infusion for 8 hr in bile fistula rats. Lymph output and esterification of [14C]cholesterol and endogenous cholesterol were measured in hourly samples. A second group of bile fistula rats was given the same bile acids as the first group but without added cholesterol or other lipid, i.e., fasting lymph fistula group. Mucosal uptake of [14C]cholesterol was studied using recovery of [14C]cholesterol from lumen and mucosa after 1-hr infusions in conscious bile fistula rats. Lymph output of [14C]cholesterol was promoted more rapidly with taurocholate than with the dihydroxy conjugates and [14C]cholesterol output differed for the three groups given dihydroxy bile acids. The mass of cholesterol in lymph, measured chemically, varied in parallel with [14C]cholesterol absorption. For fasting lymph, infusion of dihydroxy bile acids failed to produce a significant change in endogenous cholesterol output when compared with rats given saline only. Taurocholate infusion markedly increased endogenous cholesterol in lymph of fasted rats. Under all conditions where cholesterol output was stimulated, the increase could be accounted for mainly as esterified cholesterol. Mucosal uptake of [14C]cholesterol during 1-hr infusions in conscious bile fistula rats was slower with the dihydroxy bile acids than with taurocholate. The results indicate the marked effect of the number and configuration of the hydroxyl groups on the solubilizing bile acid for cholesterol absorption.


In-dopants effect on the photoluminescence properties of YVO4 nanophosphors.
The powders of Yttrium vanadate (YVO4) with In-dopants were synthesized by solid-state reactions, and X-ray diffraction (XRD), transmission electron microscopy (TEM), ultraviolet-visible (UV-vis) absorption spectra, photoluminescence (PL) spectra, and the luminescence intensity change were used to characterize the samples. The results of XRD indicated that the YVO4:In3+ samples remained in pure cubic phase. TEM illustrated that the powders mainly consisted of grains with an average size of 100 nm. Under the excitation of 320 nm, the YVO4:In3+ single-crystalline samples exhibited emission ranging from 350 to 700 nm. The emission intensity of YVO4:In3+ increased with increasing indium concentration in the lower indium concentration region until the saturated PL intensity was reached, and the strongest white fluorescence was observed when the In3+ doping concentration was 2% at 900 degrees C. The luminescent intensity of YVO4:In3+ (2%) was 9.6 times as strong as that of non-doped YVO4. The nanophosphors emit white luminescence owing to broad charge transfer in crystal lattice is due to the addition of In3+ to capture the UV radiation.


Practical aspects of preparation of the patient for the thyroid gland surgery.
The aim of the study is to present practical suggestions of how to prepare a patient for thyroid gland surgery. Widespread distribution of thyroid diseases results in referring the patients for strumectomy. Proper preparation of the patient for the operation decreases the risk of perioperative complications to a minimum. Good co-operation in the team endocrinologist-anaestesiologist-surgeon is very important for the effective treatment.


Palladium catalyzed asymmetric hydrophosphination of α,β- and α,β,γ,δ-unsaturated malonate esters - efficient control of reactivity, stereo- and regio-selectivity.
Both PC-cyclometalated and PCP-pincer type palladium catalysts have recently been found to be robust and efficacious catalysts for the asymmetric P-H addition reaction involving activated olefins. Our studies on the asymmetric P-H addition of diphenylphosphine to malonate ester and α,β,γ,δ-alkylidenemalonate ester revealed for the first time that the catalyst choice can have a dramatic impact in terms of reactivity as well as regio- and stereo-control for this asymmetric hydrofunctionalization reaction. Besides showing significantly contrasting reactivity and stereoselectivity in the hydrophosphination reaction involving malonate ester, in the case of α,β,γ,δ-alkylidenemalonate ester, a novel regiodivergent method was developed with the 1,4-adduct being obtained exclusively with the PC-catalyst while the pincer catalyst produced only the 1,6-adduct.


Conceptual disorganization weakens links in cognitive pathways: Disentangling neurocognition, social cognition, and metacognition in schizophrenia.
Disentangling links between neurocognition, social cognition, and metacognition offers the potential to improve interventions for these cognitive processes. Disorganized symptoms have shown promise for explaining the limiting relationship that neurocognition holds with both social cognition and metacognition. In this study, primary aims included: 1) testing whether conceptual disorganization, a specific disorganized symptom, moderated relationships between cognitive processes, and 2) examining the level of conceptual disorganization necessary for links between cognitive processes to break down. To accomplish these aims, comprehensive assessments of conceptual disorganization, neurocognition, social cognition, and metacognition were administered to 67 people with schizophrenia-spectrum disorders. We found that conceptual disorganization significantly moderated the relationship between neurocognition and metacognition, with links between cognitive processes weakening when conceptual disorganization is present even at minimal levels of severity. There was no evidence that conceptual disorganization-or any other specific disorganized symptom-drove the limiting relationship of neurocognition on social cognition. Based on our findings, conceptual disorganization appears to be a critical piece of the puzzle when disentangling the relationship between neurocognition and metacognition. Roles of specific disorganized symptoms in the neurocognition - social cognition relationship were less clear. Findings from this study suggest that disorganized symptoms are an important treatment consideration when aiming to improve cognitive impairments.


Biotransformation of Carboxylic Acids to Alcohols: Characterization of Thermoanaerobacter Strain AK152 and 1-Propanol Production via Propionate Reduction.
Thermoanaerobacter strains have recently gained interest because of their ability to convert short chain fatty acids to alcohols using actively growing cells. Thermoanaerobacter thermohydrosulfuricus strain AK152 was physiologically investigated for its ethanol and other alcohol formation. The temperature and pH optimum of the strain was 70 °C and pH 7.0 and the strain degraded a variety of compounds present in lignocellulosic biomass like monosaccharides, disaccharides, and starch. The strain is highly ethanologenic, producing up to 86% of the theoretical ethanol yield form hexoses. Strain AK152 was inhibited by relatively low initial substrate (30 mM) concentration, leading to inefficient degradation of glucose and levelling up of all end-product formation. The present study shows that the strain produces alcohols from most of the tested carboxylic acids, with the highest yields for propionate conversion to propanol (40.7%) with kinetic studies demonstrating that the maximum conversion happens within the first 48 h of fermentation. Various physiological tests were performed to maximize the acid conversion to the alcohol which reveals that the optimum pH for propionate conversion is pH 6.7 which affords a 57.3% conversion. Kinetic studies reveal that propionate conversion is rapid, achieving a maximum conversion within the first 48 h of fermentation. Finally, by using 13C NMR, it was shown that the addition of propionate indeed converted to propanol.


Predictors of mammography use among women veterans.
We examined mammography use patterns of women veterans and explored Veterans Administration (VA) health care use and military experience as predictors of mammography use by this population. We conducted a national telephone survey of women veterans. A sample of 397 women veterans was selected from all military discharges from 1971 through 1994. A 3 x 2 stratification scheme was used: three age groups (35-49, 50-64, > or = 65 years old) and two VA user groups indicating whether (VA user) or not (VA nonuser) they received any health care from a VA Medical Center in the last 5 years. The response rate was 75% (297 of 397). Analyses included bivariate techniques and weighted logistic regression. We found that women veterans told to have a mammogram by a health care professional were more than five times more likely to have ever had a mammogram (OR 5.41, CI 4.63-6.32) and nearly twice as likely to have had a mammogram within the past 2 years (OR 1.81, CI 1.57-2.09) as those who were not told to do so, controlling for age, race, VA user status, and length of military service. Regular VA users were more likely to have had a mammogram ever and within the past 2 years, controlling for other factors. Mammography use was not necessarily at a VA medical center. Interventions that promote better provider-patient communication and target older women veterans may have the most potential benefit. Whether VA health care is filling an important gap in access to mammography for older women veterans is an important policy question and warrants further research.


The role of SREBP-1c in nutritional regulation of lipogenic enzyme gene expression.
A high carbohydrate diet up-regulates the transcription of enzymes of triglyceride biosynthesis (lipogenesis) in the mammalian liver. This treatment stimulates hepatic insulin signaling, leading to transcription of sterol regulatory element-binding protein-1c (SREBP-1c). SREBP-1c has been implicated as a major factor that up-regulates lipogenic genes in response to carbohydrate feeding. However, we presented evidence for another factor, carbohydrate response factor, which is also involved in this response, and we proposed a model wherein SREBP-1c and carbohydrate response factor are independent transcription factors that act in response to insulin and glucose, respectively. In this study, we examined the contribution of SREBP-1c to the expression of lipogenic genes in glucose- and insulin-treated primary rat hepatocytes using an inducible adenovirus system. We found that SREBP-1c overexpression leads to a modest induction of fatty acid synthase, S(14), and acetyl-CoA carboxylase mRNAs to 20% (fatty acid synthase), 10% (S(14)), and 5% (acetyl-CoA carboxylase) of the induction seen by high glucose and insulin treatment. Restoring insulin to cells overexpressing SREBP-1c did not further increase these mRNA levels. In contrast, adenovirus-expressed SREBP-1c did not induce pyruvate kinase mRNA, suggesting that induction of this gene is SREBP-1c-independent. SREBP-1c does indeed play a role in the induction of lipogenic enzyme genes in response to insulin treatment, but it is not sufficient for the induction seen when hepatocytes are treated with insulin and high glucose.


Identification of lysine succinylation as a new post-translational modification.
Of the 20 ribosomally coded amino acid residues, lysine is the most frequently post-translationally modified, which has important functional and regulatory consequences. Here we report the identification and verification of a previously unreported form of protein post-translational modification (PTM): lysine succinylation. The succinyllysine residue was initially identified by mass spectrometry and protein sequence alignment. The identified succinyllysine peptides derived from in vivo proteins were verified by western blot analysis, in vivo labeling with isotopic succinate, MS/MS and HPLC coelution of their synthetic counterparts. We further show that lysine succinylation is evolutionarily conserved and that this PTM responds to different physiological conditions. Our study also implies that succinyl-CoA might be a cofactor for lysine succinylation. Given the apparent high abundance of lysine succinylation and the significant structural changes induced by this PTM, it is expected that lysine succinylation has important cellular functions.


The syntheses of [13 C6 ] and [phenyl-14 C(U)]BMS-816336, an inhibitor of 11β-hydroxysteroid dehydrogenase type 1, for type 2 diabetes.
Type 2 diabetes is a significant worldwide health problem. To support the development of BMS-816336 as an inhibitor of 11β-hydroxysteroid dehydrogenase type 1 for type 2 diabetes, the synthesis of carbon-14 labeled material was required for use in metabolic profiling. [Phenyl-14 C(U)]BMS-816336 was synthesized in 8 steps and 22% radiochemical yield from commercially available [14 C(U)]bromobenzene. The radiochemical purity of [phenyl-14 C(U)]BMS-816336 was 100% having a specific activity of 84.4 μCi/mg or 28.8 mCi/mmol for a total of 8.9 mCi. It was also necessary to synthesize [13 C6 ]BMS-816336 for use as a liquid chromatography/mass spectrometry standard. [13 C6 ]BMS-816336 was also prepared in 8 labeled steps in 26% yield from [13 C6 ]bromobenzene.


Validation the performance of New York Sepsis Severity Score compared with Sepsis Severity Score in predicting hospital mortality among sepsis patients.
The aim of this study was to compare the performance of the New York Sepsis Severity Score (NYSSS) with the Sepsis Severity Score (SSS) and Acute Physiology and Chronic Health Evaluation and Simplified Acute Physiology Scores for predicting mortality in sepsis patients. A retrospective analysis was conducted in the intensive care unit. The primary outcome was in-hospital mortality. Overall 1680 sepsis patients were enrolled. The hospital mortality rate was 44.4%. The NYSSS underestimated actual mortality with standard mortality ratio (SMR) of 1.28 (95%CI 1.19-1.38). However, the SSS slightly overestimated the actual mortality with an SMR of 0.94 (0.88-1.01). The NYSSS had moderate discrimination with an AUC of 0.772 (0.750-0.794), in contrast to the SSS which had good discrimination with an AUC of 0.889 (0.873-0.904). The AUC of the SSS was statistically higher than that of the NYSSS. The AUCs of both the NYSSS and SSS were significantly lower than other standard severity scores. The calibrations for all severity scores were poor. The SSS had better overall performance than the NYSSS (Brier score 0.149 and 0.201, respectively). The SSS had better discrimination and overall performance than the NYSSS. However, both sepsis severity scores were poorly calibrated.


Ethnicity as a factor in pizza delivery crashes.
A subjective assessment of patients admitted acutely to the orthopedic directorate revealed that there had been--over a short period of time--a small series of patients who had sustained fractures during the course of their employment as pizza delivery moped drivers. Consequently an observational study was performed in our district general hospital in order to investigate this further, and establish the demographic domain of the patients involved and the cost of their injuries. Our hospital has a patient catchment area of 350,000 and an emergency department that treats 95,000 patients annually. The patients in this series were described in terms of their injuries, ethnicity, and cost of injury treatment. The main findings were that pizza delivery personnel admitted acutely were male Afghans, with poor English language skills, who sustained significant fractures in moped accidents that were expensive to treat.


Morphological and biochemical correlates in the characterization of Sarcocystis spp.
Isoenzyme electrophoretic techniques were applied to the characterization of seven Sarcocystis spp. that had been identified by conventional morphological studies. Cystozoites were harvested from macroscopic cysts from sheep, cattle, and mice and from microscopic cysts from sheep, cattle, and goats. Soluble cystozoite extracts were subjected to cellulose acetate gel electrophoresis and characterized at 15 of the 39 enzyme loci examined. Genetic relationships among isolates were examined by simple phenetic clustering. Two different morphological types of macroscopic cysts from sheep, identified as S. gigantea (syn. S. ovifelis) and S. medusiformis, consistently differed at 40% of the loci examined. Such genetic divergence confirms their separate morphotypic classification. Both differed from microscopic cyst isolates from sheep at 87% of the loci examined; however, two different morphotypes of microscopic cysts were found in the sheep sampled (thick-walled and thin-walled cysts). Until sufficient numbers of each type can be isolated and examined separately, both were regarded as belonging to the species S. tenella (syn. S. ovicanis). Macroscopic and microscopic cysts from cattle consistently differed at 80% of the loci thereby supporting their separate classification as S. hirsuta (syn. S. bovifelis) and S. cruzi (syn. S. bovicanis), respectively. Isolates from goats (microscopic cysts identified as S. capracanis) differed from S. tenella and S. cruzi at 20% and 47% of the loci, respectively. All macroscopic cyst isolates from the various host animal species (including S. muris from mice) differed from each other at nearly all loci. Isoenzyme electrophoretic techniques therefore provided genetic evidence supporting the classification of these various Sarcocystis spp. by their morphological characteristics.


Effectiveness of Pulse Oximetry Versus Doppler for Tourniquet Monitoring.
Pulse oximeters are common and include arterial pulse detection as part of their methodology. The authors investigated the possible usefulness of pulse oximeters for monitoring extremity tourniquet arterial occlusion. Tactical Ratcheting Medical Tourniquets were tightened to the least Doppler-determined occluding pressure at mid-thigh or mid-arm locations on one limb at a time on all four limbs of 15 volunteers. A randomized block design was used to determine the placement locations of three pulse oximeter sensors on the relevant digits. The times and pressures of pulsatile signal absences and returns were recorded for 200 seconds, with the tourniquet being tightened only when the Doppler ultrasound and all three pulse oximeters had pulsatile signals present (pulsatile waveform traces for the pulse oximeters). From the first Doppler signal absence to tourniquet release, toe-located pulse oximeters missed Doppler signal presence 41% to 50% of the times (discrete 1-second intervals) and missed 39% to 49% of the pressure points (discrete 1mmHg intervals); fingerlocated pulse oximeters had miss rates of 11% to 15% of the times and 13% to 19% of the pressure points. On toes, the pulse oximeter ranges of sensitivity and specificity for Doppler pulse detection were 71% to 90% and 44% to 51%, and on fingers, the respective ranges were 65% to 77% and 78% to 83%. Use of a pulse oximeter to monitor limb tourniquet effectiveness will result in some instances of an undetected weak arterial pulse being present. If a pulse oximeter waveform is obtained from a location distal to a tourniquet, the tourniquet should be tightened. If a pulsatile waveform is not detected, vigilance should be maintained.


Correlation between atrial fibrillation, serum amyloid protein A and other inflammatory cytokines.
Atrial fibrillation (AF) may occur and persist due to atrial remodeling exacerbated by inflammation. However, whether inflammatory cytokines, such as serum amyloid protein A (SAA), are elevated in patients with atrial arrhythmias remains unclear. In a case-control study design, serum levels of inflammatory cytokines SAA, IL-1, IL-6, TNF-α and high-sensitivity C-reactive protein (hsCRP) were compared between 122 patients with AF and 63 non-AF controls. Of 122 AF patients, 48 had lone AF and 74 organic AF; 65 had paroxysmal AF, 36 persistent AF and 21 permanent AF. In unadjusted analyses, the levels of SAA, hsCRP, TNF-αP, IL-1 and IL-6 were significantly higher in AF patients compared to control patients (P<0.01). Patients with permanent and persistent AF had higher serum levels of inflammatory cytokines than those with paroxysmal AF; all AF patients had higher serum levels than controls (P<0.05). Using a multivariate logistic regression analysis, we found that SAA (OR 3.11; 95% CI, 1.12-6.05), hsCRP (OR 2.53; 95% CI, 1.17-4.38), TNF-α (OR 1.80; 95% CI, 1.03-3.20) and IL-6 (OR 1.21; 95% CI, 0.68-2.87) were correlated with AF (P<0.05). SAA and related inflammatory cytokines are elevated in AF patients. This study is the first to document elevated SAA in AF patients. Although the cause of elevated SAA levels in AF patients remains unknown, elevated SAA may reflect an inflammatory state that promotes the occurrence and persistence of AF.


Ion temperature anisotropy across a magnetotail reconnection jet.
A significant fraction of the energy released by magnetotail reconnection appears to go into ion heating, but this heating is generally anisotropic. We examine ARTEMIS dual-spacecraft observations of a long-duration magnetotail exhaust generated by antiparallel reconnection in conjunction with particle-in-cell simulations, showing spatial variations in the anisotropy across the outflow far (>100di ) downstream of the X line. A consistent pattern is found in both the spacecraft data and the simulations: While the total temperature across the exhaust is rather constant, near the boundaries Ti,|| dominates. The plasma is well above the firehose threshold within patchy spatial regions at |BX |∈[0.1,0.5]B0, suggesting that the drive for the instability is strong and the instability is too weak to relax the anisotropy. At the midplane ( |BX|≲0.1B0), Ti,⊥>Ti,|| and ions undergo Speiser-like motion despite the large distance from the X line.


Germ-cell tumors in patients with apparently normal testes.
Germ-cell tumors occasionally occur in patients with clinically normal testes. Although data exist correlating serial serum hCG and AFP in gem-cell neoplasms of testicular origin, the literature is sparse concerning the patients presenting with clinically normal testes. Presented in this report are data to confirm the reliability and usefulness of serial serum hCG and AFP levels in those patients with extragonadal germ-cell tumors and apparently normal testes. Six patients with nonseminomatous extragonadal tumors are presented. Serial serum hCG and AFP levels correlated with the clinical course in each of the patients. The serum hCG and/or AFP decreased with response to therapy and increased with progression of disease. It is concluded that serial serum hCG and AFP determinations are very useful parameters to evaluate and follow the course of disease in this group of patients.


Adaptation to hypoosmotic challenge in brachyuran crabs: a microanatomical and electrophysiological characterization of the intestinal epithelia.
Besides its role in digestion and nutrient absorption, the crustacean gut participates in osmo/ionic regulation. We investigate microanatomy, ionic permeability and transepithelial electrophysiological parameters in the mid- and hindguts of three hyperosmoregulating crabs that inhabit estuarine waters (Chasmagnathus granulata), brackish mangrove swamp (Sesarma rectum) or freshwater (Dilocarcinus pagei). The abdominal hindguts are cuticle lined, the single-layered epithelia consisting of narrow, columnar cells exhibiting apically dense, unvesiculated cytoplasm. In the saltwater species, the thoracic midgut epithelium consists of tall, narrow, columnar cells displaying numerous, apical microvilli above dense apical cytoplasm. However, the corresponding gut segment in the hololimnetic species, D. pagei, consists of squat cells lacking apical microvilli, overlain by a heavy cuticle, constituting a thoracic or anterior hindgut. The midgut/thoracic hindgut epithelia in all three crabs, and abdominal (posterior) hindgut of D. pagei, exhibit similar, small, lumen-negative voltages when perfused symmetrically with hemolymph-like salines. The hindguts of the saltwater species show similar, small, lumen-positive voltages. Small short-circuit currents are detectable after voltage clamping. Washout and/or addition of luminal glucose or amino acids do not alter current or conductance, suggesting the absence of active, electrogenic nutrient absorption. Ion substitution did not disclose active, electrogenic absorption or secretion of Na+ and/or Cl-. The midguts of the saltwater species exhibit similar conductances, greater than in D. pagei, but no ion selectivity; hindgut conductance is low, the epithelia showing moderate anion selectivity. The thoracic (anterior) and abdominal (posterior) hindgut epithelia of D. pagei, the freshwater species, exhibit similar, low conductances, and are ion selective. These findings reveal that active, electrogenic, salt and nutrient transport is undetectably low or absent. The reduced transepithelial conductances and notable ion selectivities in the abdominal and thoracic hindguts of D. pagei may reduce passive salt losses in fresh water, contributing to osmotic and ionic regulation.


TRP channels and lipids: from Drosophila to mammalian physiology.
The transient receptor potential (TRP) ion channel family was the last major ion channel family to be discovered. The prototypical member (dTRP) was identified by a forward genetic approach in Drosophila, where it represents the transduction channel in the photoreceptors, activated downstream of a Gq-coupled PLC. In the meantime 29 vertebrate TRP isoforms are recognized, distributed amongst seven subfamilies (TRPC, TRPV, TRPM, TRPML, TRPP, TRPA, TRPN). They subserve a wide range of functions throughout the body, most notably, though by no means exclusively, in sensory transduction and in vascular smooth muscle. However, their precise physiological roles and mechanism of activation and regulation are still only gradually being revealed. Most TRP channels are subject to multiple modes of regulation, but a common theme amongst the TRPC/V/M subfamilies is their regulation by lipid messengers. Genetic evidence supports an excitatory role of diacylglycerol (DAG) for the dTRP's, although curiously only DAG metabolites (PUFAs) have been found to activate the Drosophila channels. TRPC2,3,6 and 7 are widely accepted as DAG-activated channels, although TRPC3 can also be regulated via a store-operated mechanism. More recently PIP2 has been shown to be required for activity of TRPV5, TRPM4,5,7 and 8, whilst it may inhibit TRPV1 and the dTRPs. Although compelling evidence for a direct interaction of DAG with the TRPC channels is lacking, mutagenesis studies have identified putative PIP2-interacting domains in the C-termini of several TRPV and TRPM channels.


Association between the CHEK2*1100delC germ line mutation and estrogen receptor status.
Already published data were further analyzed regarding the association between the CHEK2*1100delC germ line mutation and estrogen receptor (ER) status in patients with breast cancer. The CHEK2*1100delC mutation was more prevalent among the patients with a positive ER status (4.2% versus 1.0%). An ER-negative status was beside CHEK2*1100delC mutation and independently associated with an earlier of age onset of breast cancer. There was a trend that an ER-negative status, beside the presence of a CHEK2*1100delC mutation, was associated with a worse disease-free survival. There might be an association between ER status and a CHEK2*1100delC mutation. More studies with larger number of patients are needed to further investigate the relation between CHEK2*1100delC and ER status.


Enhancing oligodendrocyte differentiation by transient transcription activation via DNA nanoparticle-mediated transfection.
Current approaches to derive oligodendrocytes from human pluripotent stem cells (hPSCs) need extended exposure of hPSCs to growth factors and small molecules, which limits their clinical application because of the lengthy culture time required and low generation efficiency of myelinating oligodendrocytes. Compared to extrinsic growth factors and molecules, oligodendrocyte differentiation and maturation can be more effectively modulated by regulation of the cell transcription network. In the developing central nervous system (CNS), two basic helix-loop-helix transcription factors, Olig1 and Olig2, are decisive in oligodendrocyte differentiation and maturation. Olig2 plays a critical role in the specification of oligodendrocytes and Olig1 is crucial in promoting oligodendrocyte maturation. Recently viral vectors have been used to overexpress Olig2 and Olig1 in neural stem/progenitor cells (NSCs) to induce the maturation of oligodendrocytes and enhance the remyelination activity in vivo. Because of the safety issues with viral vectors, including the insertional mutagenesis and potential tumor formation, non-viral transfection methods are preferred for clinical translation. Here we report a poly(β-amino ester) (PBAE)-based nanoparticle transfection method to deliver Olig1 and Olig2 into human fetal tissue-derived NSCs and demonstrate efficient oligodendrocyte differentiation following transgene expression of Olig1 and Olig2. This approach is potentially translatable for engineering stem cells to treat injured or diseased CNS tissues. Current protocols to derive oligodendrocytes from human pluripotent stem cells (hPSCs) require lengthy culture time with low generation efficiencies of mature oligodendrocytes. We described a new approach to enhance oligodendrocyte differentiation through nanoparticle-mediated transcription modulation. We tested an effective transfection method using cell-compatible poly (β-amino ester) (PBAE)/DNA nanoparticles as gene carrier to deliver transcription factor Olig1 and Olig2 into human fetal tissue-derived neural stem/progenitor cells, and showed efficient oligodendrocyte differentiation following transgene expression of Olig1 and Olig2. We believe that this translatable approach can be applied to many other cell-based regenerative therapies as well.


Long-Distance Spread of Verticillium dahliae Through Rivers and Irrigation Systems.
Verticillium dahliae Kleb. is a soilborne pathogen causing Verticillium wilt disease on several hosts. The pathogen survival structure (i.e., microsclerotia) can be efficiently spread by different dispersal methods. In the present study, the medium to long dispersal spread of the pathogen through rivers and irrigation canals was investigated. Samples of sediments (n = 29) were gathered from eight Lebanese rivers and three regional irrigation canals, in addition to samples of soil particles and plant residues (n = 14) from irrigation filters in commercial orchards. Specific conventional and real-time nested polymerase chain reaction assays detected the pathogen in six rivers-Al Kabir, Al Bared, Litani, Al Awali, Ostwan, and Litani South-and in all sampled canals-Ostwan, Al Bared, and Litani Canal 900. Starting DNA quantities ranged from 0.2 pg to 21.318 ng and an inoculum density, determined by a traditional plating method, varied between nondetectable and 0.2 microsclerotia/g. Viable V. dahliae microsclerotia were also found in residues collected from mesh-type irrigation filters of five commercial orchards. This study confirms that water is an important inoculum source of V. dahliae, being involved in the efficient spread of microsclerotia in Lebanese agricultural areas.


Protein evolution in different cellular environments: cytochrome b in sharks and mammals.
DNA sequences for the mitochondrial cytochrome b gene were determined for 13 species of sharks. Rates and patterns of amino acid replacement are compared for sharks and mammals. Absolute rates of cytochrome b evolution are six times slower in sharks than in mammals. Bivariate plots of the number of nonsynonymous and silent transversions are indistinguishable in the two groups, however, suggesting that the differences in amino acid replacement rates are due primarily to differences in DNA substitution rates. Patterns of amino acid replacement are also similar in the two groups. Conserved and variable regions occur in the same parts of the cytochrome b gene, and there is little evidence that the types of amino acid changes are significantly different between the groups. Similarity in the relative rates and patterns of protein change between the two groups prevails despite dramatic differences in the cellular environments of sharks and mammals. Poor penetrance of physiological differences through to rates of protein evolution provides support for the neutral theory and suggests that, for cytochrome b, patterns of evolution have been relatively constant throughout much of vertebrate history.


Lightness induction enhancements and limitations at low frequency modulations across a variety of stimulus contexts.
Lightness illusions are often studied under static viewing conditions with figures varying in geometric design, containing different types of perceptual grouping and figure-ground cues. A few studies have explored the perception of lightness induction while modulating lightness illusions continuously in time, where changes in perceived lightness are often linked to the temporal modulation frequency, up to around 2-4 Hz. These findings support the concept of a cut-off frequency for lightness induction. However, another critical change (enhancement) in the magnitude of perceived lightness during slower temporal modulation conditions has not been addressed in previous temporal modulation studies. Moreover, it remains unclear whether this critical change applies to a variety of lightness illusion stimuli, and the degree to which different stimulus configurations can demonstrate enhanced lightness induction in low modulation frequencies. Therefore, we measured lightness induction strength by having participants cancel out any perceived modulation in lightness detected over time within a central target region, while the surrounding context, which ultimately drives the lightness illusion, was viewed in a static state or modulated continuously in time over a low frequency range (0.25-2 Hz). In general, lightness induction decreased as temporal modulation frequency was increased, with the strongest perceived lightness induction occurring at lower modulation frequencies for visual illusions with strong grouping and figure-ground cues. When compared to static viewing conditions, we found that slow continuous surround modulation induces a strong and significant increase in perceived lightness for multiple types of lightness induction stimuli. Stimuli with perceptually ambiguous grouping and figure-ground cues showed weaker effects of slow modulation lightness enhancement. Our results demonstrate that, in addition to the existence of a cut-off frequency, an additional critical temporal modulation frequency of lightness induction exists (0.25-0.5 Hz), which instead maximally enhances lightness induction and seems to be contingent upon the prevalence of figure-ground and grouping organization.


Training of resistance to proactive interference and working memory in older adults: a randomized double-blind study.
Working memory (WM) performance is often decreased in older adults. Despite the growing popularity of WM trainings, underlying mechanisms are still poorly understood. Resistance to proactive interference (PI) constitutes a candidate process that contributes to WM performance and might influence training or transfer effects. Here, we investigated whether PI resistance can be enhanced in older adults using a WM training with specifically increased PI-demands. Further, we investigated whether potential effects of such a training were stable and entailed any transfer on non-trained tasks. Healthy old adults (N = 25, 68.8 ± 5.5 years) trained with a recent-probes and an n-back task daily for two weeks. Two different training regimens (high vs. low PI-amount in the tasks) were applied as between-participants manipulation, to which participants were randomly assigned. Near transfer tasks included interference tasks; far transfer tasks assessed fluid intelligence (gF) or speed. Immediate transfer was assessed directly after training; a follow-up measurement was conducted after two months. Both groups similarly improved in PI resistance in both training tasks. Thus, PI susceptibility was generally reduced in the two training groups and there was no difference between WM training with high versus low PI demands. Further, there was no differential near or far transfer on non-trained tasks, neither immediately after the training nor in the follow-up. PI-demands in WM training tasks do not seem critical for enhancing WM performance or PI resistance in older adults. Instead, improved resistance to PI appears to be an unspecific side-effect of a WM training.


Estimating preferences for modes of drug administration: The case of US healthcare professionals.
There are hidden drug administration costs that arise from a mismatch between end-user preferences and how manufacturers choose to formulate their drug products for delivery to patients. The corollary of this is: there are "intangible benefits" from considering end-user preferences in manufacturing patient-friendly medicines. It is important then to have some idea of what pharmaceutical manufacturers should consider in making patient-friendly medicines and of the magnitude of the indirect benefits from doing so. This study aimed to evaluate preferences of healthcare professionals in the US for the non-monetary attributes of different modes of drug administration. It uses these preference orderings to compute a monetary valuation of the indirect benefits from making patient-friendly medicines. A survey collected choice preferences of a sample of 210 healthcare professionals in the US for two unlabelled drug options. These drugs were identical except in the levels of attributes of drug administration. Using the choice data collected, statistical models were estimated to compute gross welfare benefits, measured by the expected compensating variation, from making drugs in a more patient-friendly manner. The monetary value of end-user benefits from developing patient-friendly drug delivery systems is: (1) as large as the annual acquisition costs per full treatment episode for some biologic drugs; and (2) likely to fall in the "high end" of the distribution of the direct monetary costs of drug administration. An examination of end-user preferences should help manufacturers make more effective and efficient use of limited resources for innovations in drug delivery system, or manufacturing research in general.


Comparative analysis of diagnostic procedures for tumor detection rates in paired data.
Diagnostic procedures are mostly used to detect a particular disease, and each procedure indicates the presence or absence of the disease in an individual. Sensitivity and positive predictive value, which are measures of the effectiveness of a diagnostic procedure, are simply calculated as the proportion of the individuals diagnosed with the disease by the test among the patients with the disease, and of the diseased persons among the individuals in whom the disease was detected by the test, respectively. For a diagnosis with such a binary result, sensitivity and the positive predictive value of diagnostic procedures can be compared using the chi-square statistic. However, in the treatment of cancer patients, it is important not only to diagnose the disease status of an individual patient but also to detect the correct location of the cancer. The tumor location may be incorrectly identified in some subjects diagnosed with cancer. It is therefore of interest whether a procedure that diagnoses cancer also correctly indicates the tumor location. In this paper, we re-define the sensitivity and the positive predictive value of tumor detection as the ratio of the number of cases with a correct diagnosis of the tumor location by the test to the number of cases of cancer, and as the ratio of patients with a correct diagnosis of the tumor location to the number of individuals diagnosed with cancer by the test, respectively. We refer to these parameters as 'semi-sensitivity' and 'semi-positive predictive value'. To compare these ratios between diagnostic procedures, test statistics are developed from binary diagnostic results. Simulation studies conducted to evaluate the nominal level and power are presented, and two sets of example data are also analyzed using the new test statistic.