Enhancing response of a protein conformational switch by using two disordered ligand binding domains.
Introduction: Protein conformational switches are often constructed by fusing an input domain, which recognizes a target ligand, to an output domain that establishes a biological response. Prior designs have employed binding-induced folding of the input domain to drive a conformational change in the output domain. Adding a second input domain can in principle harvest additional binding energy for performing useful work. It is not obvious, however, how to fuse two binding domains to a single output domain such that folding of both binding domains combine to effect conformational change in the output domain. Methods: Here, we converted the ribonuclease barnase (Bn) to a switchable enzyme by duplicating a C-terminal portion of its sequence and appending it to its N-terminus, thereby establishing a native fold (OFF state) and a circularly permuted fold (ON state) that competed for the shared core in a mutually exclusive fashion. Two copies of FK506 binding protein (FKBP), both made unstable by the V24A mutation and one that had been circularly permuted, were inserted into the engineered barnase at the junctions between the shared and duplicated sequences. Results: Rapamycin-induced folding of FK506 binding protein stretched and unfolded the native fold of barnase via the mutually exclusive folding effect, and rapamycin-induced folding of permuted FK506 binding protein stabilized the permuted fold of barnase by the loop-closure entropy principle. These folding events complemented each other to turn on RNase function. The cytotoxic switching mechanism was validated in yeast and human cells, and in vitro with purified protein. Discussion: Thermodynamic modeling and experimental results revealed that the dual action of loop-closure entropy and mutually exclusive folding is analogous to an engine transmission in which loop-closure entropy acts as the low gear, providing efficient switching at low ligand concentrations, and mutually exclusive folding acts as the high gear to allow the switch to reach its maximum response at high ligand concentrations.


Auditory dysfunction accompanying noise-induced hearing loss.
Two groups of 16 young male subjects with normal low- and midfrequency hearing were compared on a series of audiometric measures. One group was composed of subjects with 12- to 24-month histories of noise exposure and hearing loss at 4000 Hz greater than 40 dB; the other group was composed of normal-hearing subjects with no history of unusual noise exposure. On fixed-frequency Bekesy audiometry at 2000 Hz, 12 of the noise-exposed subjects demonstrated separation of 5 dB or more between pulsed- and continuous-tone tracings; similar separation occurred for only one of the non-noise-exposed subjects. Significant between-group differences also occurred on three tests of speech discrimination: PB-50 word lists and CID W-22 lists presented with two forms of competing noise.


[Treatment of gastric cancer based on the pathological and biophysiological aspects of cancer].
Spread of the gastric cancer is multifarious according to its pathological and biophysiological characteristics. The therapeutic strategy was reviewed by the analysis of more than 8.000 cases of gastric cancer treated at Cancer Institute Hospital. Main type of recurrence in early stage cancer was, though seldom, the hematogenous metastasis to the liver. On the other hand, in the advanced cases, macroscopically localized, and histologically well differentiated tumor type easily adhered and invaded to the adjacent organs and structures, and infiltrating and undifferentiated tumor type tended to cause peritoneal dissemination. Lymph node spread was observed in any tumor type, but frequently observed in intermediate type. Invasion to the adjacent organs and limited lymphatic spread can be controlled by wide combined resection of adjacent organs, and extended lymphadenectomy. However, hematogenous metastasis and peritoneal dissemination are difficult to be controlled by surgical intervention. Thus, minor surgery can be directed for small early stage cancer under strict criteria, and wide and extended resection for localized type, and combination of surgery and adjuvant chemotherapy for intermediate and infiltrating tumor types advancing more than stage II.


Elucidation of apparent non-maternity with DNA probes detecting highly polymorphic single locus systems.
During paternity testing, we encountered the following constellation in the Jk system: the mother's phenotype was Jk(a-b+), while the son was typed as Jk(a+b-). The deduced genotype of the mother would have been Jkb Jkb, and each offspring should then express the Jk(b) antigen. Consequently, non-maternity would be deduced. Since no material was available for extended family studies or HLA typing, except for the DNA of the propositi, only RFLP analysis could bring clarification in this case. The application of four highly polymorphic single locus probes proved the maternity and hence the existence of a Jk-Null allele. We conclude that direct testing at the DNA level may help resolving cases where, by conventional parentage testing, conclusive results are unachievable because of putative 'Null' alleles.


In vivo treatment with anti-ICAM-1 and anti-LFA-1 antibodies inhibits contact sensitization-induced migration of epidermal Langerhans cells to regional lymph nodes.
Development of contact hypersensitivity in mice depends on the migration of Langerhans cells from the epidermis to regional lymph nodes. Since ICAM-1 and LFA-1 play important roles in leukocyte migration, we sought to determine whether in vivo administration of anti-ICAM-1 and anti-LFA-1 antibodies would inhibit contact sensitization-induced migration of epidermal Langerhans cells to regional lymph nodes. Twenty-four hours after contact sensitization of mice with FITC, a brightly FITC-stained Ia+ population of dendritic cells capable of stimulating a FITC-specific Ia-restricted T-cell hybridoma was readily detected in their draining lymph nodes. Animals treated with anti-Ia mAb, which depletes Ia+ cells in lymph nodes and spleen but not Ia+ Langerhans cells in the epidermis, had normal numbers of FITC-bearing Ia+ cells capable of stimulating the T-cell hybridoma. Dendritic lymph node cells from mice treated with anti-ICAM-1 and anti-LFA-1 mAb were devoid of brightly FITC-stained cells and cells capable of stimulating the FITC-specific T-cell hybridoma. The combination of anti-ICAM-1 and anti-LFA-1 mAb completely inhibited the induction of contact hypersensitivity to FITC. Animals treated with anti-ICAM-1 or anti-LFA-1 monoclonal antibodies alone had significantly reduced (by 79 and 36%, respectively) numbers of brightly stained cells capable of stimulating the hybridomas. These data suggest that the adhesion molecules, ICAM-1 and LFA-1, play a significant role in contact hypersensitivity-induced migration of Langerhans cells to regional lymph nodes. The immunomodulatory effects of anti-adhesion molecule antibodies in vivo may be in part due to their effects on antigen-presenting cell migration.


Long-term effect of indomethacin on a rat model of neonatal hypoxia ischemic encephalopathy through behavioral tests.
Many medical experts prescribe indomethacin because of its anti-inflammatory, analgesic, tocolytic, and duct closure effects. This article presents an evaluation of the enduring impact of indomethacin on neonatal rats with hypoxic-ischemic (HI) insults, employing behavioral tests as a method of assessment. The experiment was conducted on male Wistar-Albino rats weighing 10 to 15 g, aged between seven and 10 days. The rats were divided into three groups using a random allocation method as follows: hypoxic ischemic encephalopathy (HIE) group, HIE treated with indomethacin group (INDO), and Sham group. A left common carotid artery ligation and hypoxia model was applied in both the HIE and INDO groups. The INDO group was treated with 4 mg/kg intraperitoneal indomethacin every 24 h for 3 days, while the Sham and HIE groups were given dimethylsulfoxide (DMSO). After 72 h, five rats from each group were sacrificed and brain tissue samples were stained with 2,3,5-Triphenyltetrazolium chloride (TCC) for infarct-volume measurement. Seven rats from each group were taken to the behavioral laboratory in the sixth postnatal week (PND42) and six from each group were sacrificed for the Evans blue (EB) experiment for blood-brain barrier (BBB) integrity evaluation. The open field (OF) test and Morris water maze (MWM) tests were performed. After behavioral tests, brain tissue were obtained and stained with TCC to assess the infarct volume. The significant increase in the time spent in the central area and the frequency of crossing to the center in the INDO group compared with the HIE group indicated that indomethacin decreased anxiety-like behavior (p < 0.001, p < 0.05). However, the MWM test revealed that indomethacin did not positively affect learning and memory performance (p > 0.05). Additionally, indomethacin significantly reduced infarct volume and neuropathological grading in adolescence (p < 0.05), although not statistically significant in the early period. Moreover, the EB experiment demonstrated that indomethacin effectively increased BBB integrity (p < 0.05). In this study, we have shown for the first time that indomethacin treatment can reduce levels of anxiety-like behavior and enhance levels of exploratory behavior in a neonatal rat model with HIE. It is necessary to determine whether nonsteroidal anti-inflammatory agents, such as indomethacin, should be used for adjuvant therapy in newborns with HIE.


Stable correction of antimongoloid deformity of the external canthus.
The antimongoloid deformity comprises inferior dystopia of the lateral canthus. It causes a sad looking appearance, inferior scleral show and laxity of the lower eyelid. Many different procedures have been described to correct this deformity. We believe that a stable and lasting correction can be achieved using a coronal-approach subperiosteal mask-lift, combined with DMAS (Deep Musculo-Aponeurotic System) suspension of a temporo-masseteric anchor-flap (instead of a simple canthoplasty). If bony augmentation of the cheek bones is required, we use an autograft of cartilage of the concha of the ear. While this 'orthomorphic' procedure is primarily applied in esthetic cosmetic surgery, it can also be used to correct deformities in reconstructive surgery.


Misorientation distributions in hot deformed NaCl using electron backscattered diffraction.
EBSD orientation mapping has been used to derive subgrain boundary misorientation distributions in a series of hot deformed and etched NaCl samples. The main objective of this study has been to examine the influence of data processing, noise caused by angular resolution limits and step size on the subgrain misorientation distributions in hot deformed NaCl. Processing of non-indexed EBSD patterns increased the average misorientations in etched NaCl. Noise contributed significantly to low angle misorientation peaks for step sizes less than the minimum subgrain size. Orientation data collected using a step size larger than the average subgrain size cumulated misorientations across individual subgrains and effectively measured an orientation gradient between steps. Orientation gradient distributions were not influenced by noise. Average misorientation values calculated from large step data correlated well with average misorientation from small step size data, Average misorientations showed a power law relationship with strain. Three types of substructures were identified using scanning electron microscopy and EBSD mapping, equiaxed subgrains, long subgrain boundaries and a core-mantle subgrain arrangement.


Peak oxygen uptake in relation to total heart volume discriminates heart failure patients from healthy volunteers and athletes.
An early sign of heart failure (HF) is a decreased cardiac reserve or inability to adequately increase cardiac output during exercise. Under normal circumstances maximal cardiac output is closely related to peak oxygen uptake (VO2peak) which has previously been shown to be closely related to total heart volume (THV). Thus, the aim of this study was to derive a VO2peak/THV ratio and to test the hypothesis that this ratio can be used to distinguish patients with HF from healthy volunteers and endurance athletes. Thirty-one patients with HF of different etiologies were retrospectively included and 131 control subjects (60 healthy volunteers and 71 athletes) were prospectively enrolled. Peak oxygen uptake was determined by maximal exercise test and THV was determined by cardiovascular magnetic resonance. The VO2peak/THV ratio was then derived and tested. Peak oxygen uptake was strongly correlated to THV (r2 = 0.74, p < 0.001) in the control subjects, but not for the patients (r2 = 0.0002, p = 0.95). The VO2peak/THV ratio differed significantly between control subjects and patients, even in patients with normal ejection fraction and after normalizing for hemoglobin levels (p < 0.001). In a multivariate analysis the VO2peak/THV ratio was the only independent predictor of presence of HF (p < 0.001). The VO2peak/THV ratio can be used to distinguish patients with clinically diagnosed HF from healthy volunteers and athletes, even in patients with preserved systolic left ventricular function and after normalizing for hemoglobin levels.


Identification of a necroptosis-related gene signature as a novel prognostic biomarker of cholangiocarcinoma.
Cholangiocarcinoma (CHOL) is the most prevalent type of malignancy and the second most common form of primary liver cancer, resulting in high rates of morbidity and mortality. Necroptosis is a type of regulated cell death that appears to be involved in the regulation of several aspects of cancer biology, including tumorigenesis, metastasis, and cancer immunity. This study aimed to construct a necroptosis-related gene (NRG) signature to investigate the prognosis of CHOL patients using an integrated bioinformatics analysis. CHOL patient data were acquired from the Gene Expression Omnibus (GEO) (GSE89748, GSE107943) and The Cancer Genome Atlas (TCGA) databases, with NRGs data from the necroptosis pathway in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Univariate and multivariate regression analyses were performed to establish the NRG signatures. Kaplan-Meier (KM) curves were used to evaluate the prognosis of patients with CHOL. Functional enrichment analysis was performed to identify key NRG-associated biological signaling pathways. We also applied integrative multi-omics analysis to the high- and low-risk score groups. Spearman's rank correlation was used to clarify the relationship between the NRG signature and immune infiltration. 65 differentially expressed (DE) NRGs were screened, five of which were selected to establish the prognostic signature of NRGS based on multivariate Cox regression analysis. We observed that low-risk patients survived significantly longer than high-risk patients. We found that patients with high-risk scores experienced higher immune cell infiltration, drug resistance, and more somatic mutations than patients with low-risk scores. We further found that sensitivities to GW843682X, mitomycin C, rapamycin, and S-trityl-L-cysteine were significantly higher in the low-risk group than in the high-risk group. Finally, we validated the expression of five NRGs in CHOL tissues using the TCGA database, HPA database and our clinical data. These findings demonstrate that the five-NRG prognostic signature for CHOL patients is reasonably accurate and valid, and it may prove to be of considerable value for the treatment and prognosis of CHOL patients in the future.


Zero-based print journal collection development in a community teaching hospital library: planning for the future.
The paper describes and evaluates the success of a zero-based collection development approach to print serials in a community teaching hospital. The authors first assessed the environmental factors that would determine future needs of the medical library and its customers. Liaisons to various departments and constituencies were substantially involved in the data-gathering phase. Using newly defined collection parameters, a list of journals to consider was compiled and each journal was categorized justifying its inclusion. Any title not having a strong fit in at least one category was eliminated from further consideration. Overall, 21 subscriptions were cancelled and 34 were added. Despite a 15% increase in total subscription costs, mostly due to normal annual journal price increases, the average cost per journal went down from $344 to $327. Journal usage went up over 30%, interlibrary loan lending went down 25%, and borrowing went up 20%. As resources available to libraries decline, it becomes critical that collections and services are continually and systematically reviewed with a view to keeping them aligned with the mission of the organization, needs of the customers, and emerging trends. Zero-based collection development can be a valuable tool in bringing a print journal collection into closer alignment with the needs of library customers.


Thermostability of soluble and immobilized alpha-amylase from Bacillus licheniformis.
In view of a possible application of the alpha-amylase from Bacillus licheniformis as a time-temperature integrator for evaluation of heat processes,(11) thermal inactivation kinetics of the dissolved and covalently immobilized enzyme were studied in the temperature range 90-108 degrees C. The D-values (95 degrees C) for inactivation of alpha-amylase, dissolved in tris-HCl buffer, ranged from 6 to 157 min, depending on pH, ionic strength, and Ca(2+) and enzyme concentration. The z-value fluctuated between 6.2 and 7.6 degrees C. On immobilization of the alpha-amylase by covalent coupling with glutaraldehyde to porous glass beads, the thermoinactivation kinetics became biphasic under certain circumstances. For immobilized enzyme, the D-values (95 degrees C) ranged between 17 and 620 min, depending largely on certain environmental conditions. The z-value fluctuated between 8.1 and 12.9 degrees C. In each case of biphasic inactivation, the z-value of the stable fraction (with the higher D-values) was lower than the z-value of the labile fraction.


The evolution of a cardiac catheterization laboratory fellowship.
This column explores the development and implementation of a cardiac catheterization laboratory nursing fellowship at a 600-bed, tertiary-care hospital that is part of a large, multi-facility health system.


Green fluorescent protein as a real time quantitative reporter of heterologous protein production.
Since its cloning and commercial availability, applications of green fluorescent protein (GFP) as a reporter gene have become prevalent in many aspects of science. The attributes of GFP could also be applied to the area of heterologous protein production. The work described here represents the first experiments to use GFP as a generic tool to monitor protein production in bioprocess development. We have constructed a plasmid containing an operon fusion of the two reporter genes GFP and chloramphenicol acetyl transferase (CAT). CAT served as a "model" recombinant protein product to demonstrate the in situ quantifiable reporting mechanism of GFP. Our results indicate there is a direct correlation between the fluorescence intensity of GFP and the functional activity of the downstream CAT protein. In addition, there is a quantitative relationship between the level of CAT protein concentration and GFP fluorescence. These experiments provide the groundwork for using GFP as an in situ reporter gene for scale-up and process optimization of recombinant protein production.


Strong alkalinization in the anterior midgut of larval yellow fever mosquitoes (Aedes aegypti): involvement of luminal Na+/K+-ATPase.
Recently, Na(+)/K(+)-ATPase has been detected in the luminal membrane of the anterior midgut of larval yellow fever mosquitoes (Aedes aegypti) with immunohistochemical techniques. In this study, the possible involvement of this ATPase in strong alkalinization was investigated on the level of whole larvae, isolated and perfused midgut preparations and on the molecular level of the Na(+)/K(+)-ATPase protein. Ouabain (5 mM) did not inhibit the capability of intact larval mosquitoes to alkalinize their anterior midgut. Also in isolated and perfused midgut preparations the perfusion of the lumen with ouabain (5 mM) did not result in a significant change of the transepithelial voltage or the capacity of luminal alkalinization. Na(+)/K(+)-ATPase activity was completely abolished when KCl was substituted with choline chloride, suggesting that the enzyme cannot act as an ATP-driven Na(+)/H(+)-exchanger. Altogether the results of the present investigation indicate that apical Na(+)/K(+)-ATPase is not of direct importance for strong luminal alkalinization in the anterior midgut of larval yellow fever mosquitoes.


Gout and hyperuricemia.
Gout continues to be a health problem around the world despite the availability of effective therapies. Although the prevalence is influenced by genetic factors, the associations of alcohol consumption, obesity, and hypertension appear to be partially responsible for the increased prevalence of gout and hyperuricemia in African and Oriental countries. The association between hyperuricemia and cardiovascular disease seems linked to insulin resistance. This relation, in part, explains the common coexistence of hyperlipidemia and glucose intolerance in patients with gout. Accordingly, it is recommended that one pay more attention to dietary manipulation in patients with gout in addition to managing hypertension, obesity, and other medical problems. Although acute gout attacks can be treated, eliminating gout requires effective removal of urate from the body. Allopurinol remains a dominant urate-lowering agent, however its use may be limited by allergic reactions. Uricosuric agents are also effective urate-lowering agents and provide an alternative to allopurinol. Strategies to treat patients who are sensitive to allopurinol continue to evolve.


Age and criminal poisonings.
We present a series of 8 cases of acute combined poisonings, occurred in an identical way in patients over 70 years of age for a period of 6 months. The way of exposure, characteristic of the clinical presentation, complications and the outcome of the intoxications, as well as the therapeutic approach is described. In all of the cases combined drug intoxication with benzodiazepines and opiates have been proved. The impact of the combination of two toxic substances: the first causing rapid and brief suppression of the consciousness and the second, causing prolonged continuation of the already suppressed consciousness, on the clinical course is discussed. The similarities in the circumstances of the exposure, clinical course of the poisonings, the identified toxic substances, lead to the consideration of criminal characteristic of the poisonings. The contact with the corresponding authorities brought off the disclosure of a group of criminals, committed the intentional intoxications with the aim of robbery. Age, with all its various characteristics, has been discussed as a factor for occurrence of criminal poisonings.


Effects of residual zinc compounds and chain-end structure on thermal degradation of poly(epsilon-caprolactone).
Effects of chain-end structure and residual metal compounds on thermal degradation of poly(epsilon-caprolactone) (PCL) were investigated by means of thermogravimetric and pyrolysis-gas chromatograph mass spectrometric analyses. Four types of PCL samples with different chain-end structures (alpha-carboxylic acid-omega-hydroxyl-PCL, alpha-dodecyl ester-omega-hydroxyl-PCL, alpha-carboxylic acid-omega-acetyl-PCL, and alpha-dodecyl ester-omega-acetyl-PCL) were prepared by ring-opening polymerization of epsilon-caprolactone in the presence of zinc-based catalyst and by subsequent acetylation reaction of polymers with acetic anhydride. PCL samples with different zinc contents were obtained by washing with acetic acid in chloroform solution of polymer. Thermal degradation behaviors of these PCL samples with different chain-end structures were examined under both isothermal and nonisothermal conditions. From both the isothermal and nonisothermal experiments, the thermal degradation of PCL samples containing high amounts of residual zinc compounds from synthesis process revealed the selective unzipping depolymerization step below 300 degrees C producing epsilon-caprolactone exclusively. In contrast, zinc-free PCL samples were stable at temperatures below 300 degrees C, and the thermal degradation occurred only at temperatures above 300 degrees C regardless of the chain-end structure. From (1)H NMR analysis of the residual molecules after isothermal degradation of zinc-free PCL, it was confirmed that the omega-chain-end of residual molecules was 5-hexenoic acid unit. However, the cyclic monomer and oligomers were detected as the volatile products of zinc-free PCL samples. These results suggest that the dominant reaction of thermal degradation for PCL above 300 degrees C is a competition between the random chain scission via cis elimination reaction and the cyclic rupture via intramolecular transesterification of PCL molecules.


[Ca2+]i in single isolated cardiac cells: a review of recent results obtained with digital imaging microscopy and fura-2.
The use of fluorescent Ca2+ indicators to observe [Ca2+]i transients in voltage-clamped single cells has many advantages over previous methods, such as the use of aequorin in multicellular preparations, for studying excitation-contraction coupling. In the studies reviewed in this article, [Ca2+]i in single isolated mammalian ventricular myocytes was observed through the use of the fluorescent Ca2+ indicator, fura-2. Individual cells, loaded with fura-2 either by internal perfusion or by exposure to fura-2/AM, were generally studied with the use of inverted microscopes equipped with ultraviolet epifluorescence illumination, intensified silicon intensifier target cameras (ISIT), and (or) a photomultiplier tube. Analysis of subcellular patterns of fura-2 fluorescence was performed by digital analysis of the images obtained with the ISIT camera. Variation of membrane voltage and exposure of cells to ryanodine (which was assumed to selectively block the release of Ca2+ from the sarcoplasmic reticulum) were used to investigate the cellular processes that determine the [Ca2+]i transient. The main results of these studies are the following. (1) In any population of enzymatically isolated heart cells, there are (i) mechanically quiescent cells in which [Ca2+]i is spatially uniform, constant over time, and relatively low; (ii) spontaneously contracting cells, which have a relatively elevated [Ca2+]i, but in which the spatial uniformity of [Ca2+]i is interrupted periodically by spontaneous, propagating waves of high [Ca2+]i; and (iii) cells that are hypercontracted (rounded up) and that have higher levels of [Ca2+]i than the other two types.(ABSTRACT TRUNCATED AT 250 WORDS)


Global disparity in the supply of commercial weather and climate information services.
Information about weather and climate is vital for many areas of decision-making, particularly under conditions of increasing vulnerability and uncertainty related to climate change. We have quantified the global commercial supply of weather and climate information services. Although government data are sometimes freely available, the interpretation and analysis of those data, alongside additional data collection, are required to formulate responses to specific challenges in areas such as health, agriculture, and the built environment. Using transactional data, we analyzed annual spending by private and public organizations on commercial weather and climate information in more than 180 countries by industrial sector, region, per capita, and percentage of GDP (gross domestic product) and against the country's climate and extreme weather risk. There are major imbalances regarding access to these essential services between different countries based on region and development status. There is also no relationship between the level of climate and weather risks that a country faces and the level of per capita spending on commercial weather and climate information in that country. At the international level, action is being taken to improve access to information services. With a better understanding of the flows of commercial weather and climate information, as explored in this study, it will be possible to tackle these regional and development-related disparities and thus to increase resilience to climate and weather risks.


Vitamin A during pregnancy.
Vitamin A is an essential micronutrient throughout life. In areas of developing countries where vitamin A deficiency is endemic, an estimated 40% of the children are likely to be sub clinically deficient of the vitamin. Yet, the traditional view that preschool age children represent the main population at risk of vitamin A deficiency has been replaced by the growing awareness that sub clinical and even clinical vitamin A deficiency also occurs in women of reproductive age and infants less than 6 month of age. During the period of early foetal development the supply of vitamin A must be carefully managed to ensure that the developing foetus is exposed to neither too little not too much vitamin A because either condition can have teratogenic consequences. Towards the end of gestation, adequate maternal vitamin A status and dietary intakes are important to maximize the vitamin A transferred to the foetus in preparation for parturition and lactation. A review of the current knowledge and prospects for future research is presented.


Ontogenetic changes in the olfactory antennules of the shore crab, Hemigrapsus oregonensis, maintain sniffing function during growth.
Malacostracan crustaceans capture odours using arrays of chemosensory hairs (aesthetascs) on antennules. Lobsters and stomatopods have sparse aesthetascs on long antennules that flick with a rapid downstroke when water flows between the aesthetascs and a slow return stroke when water is trapped within the array (sniffing). Changes in velocity only cause big differences in flow through an array in a critical range of hair size, spacing and speed. Crabs have short antennules bearing dense arrays of flexible aesthetascs that splay apart during downstroke and clump together during return. Can crabs sniff, and when during ontogeny are they big enough to sniff? Antennules of Hemigrapsus oregonensis representing an ontogenetic series from small juveniles to adults were used to design dynamically scaled physical models. Particle image velocimetry quantified fluid flow through each array and showed that even very small crabs capture a new water sample in their arrays during the downstroke and retain that sample during return stroke. Comparison with isometrically scaled antennules suggests that reduction in aesthetasc flexural stiffness during ontogeny, in addition to increase in aesthetasc number and decrease in relative size, maintain sniffing as crabs grow. Sniffing performance of intermediate-sized juveniles was worse than for smaller and larger crabs.


Diurnal changes in seawater carbonate chemistry speciation at increasing atmospheric carbon dioxide.
Natural variability in seawater pH and associated carbonate chemistry parameters is in part driven by biological activities such as photosynthesis and respiration. The amplitude of these variations is expected to increase with increasing seawater carbon dioxide (CO2) concentrations in the future, because of simultaneously decreasing buffer capacity. Here, we address this experimentally during a diurnal cycle in a mesocosm CO2 perturbation study. We show that for about the same amount of dissolved inorganic carbon (DIC) utilized in net community production diel variability in proton (H+) and CO2 concentrations was almost three times higher at CO2 levels of about 675 ± 65 in comparison with levels of 310 ± 30 μatm. With a simple model, adequately simulating our measurements, we visualize carbonate chemistry variability expected for different oceanic regions with relatively low or high net community production. Since enhanced diurnal variability in CO2 and proton concentration may require stronger cellular regulation in phytoplankton to maintain respective gradients, the ability to adjust may differ between communities adapted to low in comparison with high natural variability.


STREPTOMYCIN FORMATION BY INTACT MYCELIUM OF STREPTOMYCES GRISEUS.
Nomi, Ryosaku (Rutgers, The State University, New Brunswick, N.J.). Streptomycin formation by intact mycelium of Streptomyces griseus. J. Bacteriol. 86:1220-1230. 1963.-A study was made of streptomycin formation by intact mycelium of Streptomyces griseus 107 grown in glucose-yeast extract medium. When mycelium harvested after 24, 48, and 72 hr was compared, the earliest growth showed the highest activity in producing streptomycin from glucose. The concentration of streptomycin in the mycelium was higher in the older growth. Calcium chloride had a remarkable effect in increasing streptomycin production from the precursors in the mycelium, especially when the mycelium was grown for 48 hr or longer. The effect of calcium chloride cannot be attributed to the precipitation of an excess of inorganic phosphate in the medium. Glucose, fructose, glycerol, lactic acid, glucosamine, streptidine, and inositol stimulated streptomycin formation, whereas gluconic acid, glucuronic acid, streptamine, and strepturea did not. When 24-hr-old mycelium was suspended and shaken in 0.5% glucose solution, the antibiotic precursors necessary to produce streptomycin were found mainly in the supernatant of the culture rather than in the mycelium. The supernatant included some substance which had a weak antibiotic activity. This substance was less basic than streptomycin and was transformed to streptomycin with a remarkable increase in antibiotic activity.


Investigation of L-leucine in reducing the moisture-induced deterioration of spray-dried salbutamol sulfate power for inhalation.
The aim of this study was to investigate the ability of L-leucine (LL) in preventing moisture-induced deterioration in the in vitro aerosolization performance of spray-dried (SD) salbutamol sulfate (SS). Increasing mass fraction of LL (5-80%) were co-spray dried with SS, and the physicochemical properties of the powders were characterized by laser diffraction, X-ray powder diffraction (XRD) and dynamic vapour sorption (DVS). Furthermore, the surface morphology and chemistry of fine particles was analyzed by scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The in vitro aerosolization performance of powders stored at different relative humidity (RH) was evaluated by a next generation impactor (NGI). The SD SS powders were moderately hygroscopic and amorphous, of which the uptake of moisture upon storage caused a drop in the aerosolization performance. The results showed that 40% (w/w) LL was sufficient to eliminate the effect of moisture on the aerosolization performance at 60% RH. The formulation containing 40% (w/w) LL also maximized the aerosolization performance of SD SS powders (stored in desiccator) with the emitted fraction being 90.0±1.8%, and the fine particle fraction based on the recovered dose (FPFrecovered) and emitted dose (FPFemitted) being 78.0±3.7% and 86.6±2.9%, respectively. The underlying mechanisms were that the crystalline LL increased the degree of particle surface corrugation, and reduced particle fusion and cohesiveness to facilitate dispersion. However, there is still a great challenge to prevent the moisture-induced deterioration in the aerosolization performance at 75% RH due to the recrystallization of SD SS. In conclusion, LL is a potential excipient for reducing moisture-induced deterioration in the aerosolization performance of SD amorphous powders, but still has drawbacks in preventing the recrystallization-induced deterioration.


Relationship between Structure and Properties of Nonstoichiometric Protic Ionic Liquids: n-Butylammonium Butyrate System.
Nonstoichiometric protic ionic liquids have drawn much attention in applications, including fuel cells, batteries, and reaction media. An understanding of the relationship between their structure and properties is instructive for further applications. However, there are only a few studies on nonstoichiometric protic ionic liquids. Herein, the density, viscosity, and conductivity of nonstoichiometric n-butylammonium butyrate protic ionic liquids were measured, and we used small/wide-angle scattering (S/WAXS), electron paramagnetic resonance (EPR), and molecular dynamics (MD) simulation to explore the effect of mesostructure on their properties. It is found that the hydrogen bonds drive excess N-butyric acid (PrCOOH) molecules to wrap around ion clusters, resulting in the higher density and viscosity of PrCOOH-rich PILs. The microenvironments around various radicals differ significantly in BuNH2-rich and PrCOOH-rich PILs because of the distinct molecular arrangements. This research provided a link between the physicochemical properties and structures of nonstoichiometric PILs, which is essential for their applications in electrolytes and organic reactions.


Characterization of phthalides in Ligusticum chuanxiong by liquid chromatographic-atmospheric pressure chemical ionization-mass spectrometry.
High-performance liquid chromatography (HPLC) with diode-array detection interfaced to atmospheric pressure chemical ionization (APCI)-mass spectrometry (MS) is applied to analyze phthalides from Chuanxiong (the rhizome of Ligusticum chuanxiong). This herb material, containing plenty of phthalide compositions, is selected as the analytical target in this paper for its hematological activity. Some of the phthalides are not stable and are difficult to analyze by gas chromatography-MS. Under optimized LC-MS-MS conditions, six phthalides in the methanol extract of Chuanxiong are unambiguously identified, and characteristic fragments are obtained using homemade reference standards. Ten other phthalides in the extract are confirmed by means of LC-APCI-MS with positive-negative ion mode and collision-induced dissociation in combination with UV spectrophotometry. The results show that LC-MS-MS is a method of choice for fast detection and detailed structural analysis of such mixtures in the crude extract of Chuanxiong.


Ulcerative tracheobronchitis years after colectomy for ulcerative colitis.
Two men with severe ulcerative colitis developed ulcerative tracheobronchitis 4 and 8 years after total colectomy. Intense plasma cell infiltration of tracheal mucosa and submucosa and destruction of mucous glands occurred, with partial relief of symptoms with corticosteroids. We compare them with the only other case reported, also years after colectomy.


A network involving Rho-type GTPases, a paxillin and a formin homologue regulates spore length and spore wall integrity in the filamentous fungus Ashbya gossypii.
Fungi produce spores that allow for their dispersal and survival under harsh environmental conditions. These spores can have an astonishing variety of shapes and sizes. Using the highly polar, needle-shaped spores of the ascomycete Ashbya gossypii as a model, we demonstrated that spores produced by this organism are not simple continuous structures but rather consist of three different segments that correlate with the accumulation of different materials: a rigid tip segment, a more fragile main spore-compartment and a solid tail segment. Little is currently known about the regulatory mechanisms that control the formation of the characteristic spore morphologies. We tested a variety of mutant strains for their spore phenotypes, including spore size, shape and wall defects. The mutants that we identified as displaying such phenotypes are all known for their roles in the regulation of hyphal tip growth, including the formin protein AgBni1, the homologous Rho-type GTPases AgRho1a and AgRho1b and the scaffold protein AgPxl1. Our observations suggest that these proteins form a signalling network controlling spore length by regulating the formation of actin structures.


Integrated Safety Analysis of Abrocitinib for the Treatment of Moderate-to-Severe Atopic Dermatitis From the Phase II and Phase III Clinical Trial Program.
Pivotal phase III studies demonstrated that abrocitinib, an oral, once-daily, JAK1-selective inhibitor, is effective treatment for moderate-to-severe atopic dermatitis (AD) as monotherapy and in combination with topical therapy. The aim of this study was to evaluate the long-term safety of abrocitinib 200 mg and 100 mg in an integrated analysis of a phase IIb study, four phase III studies, and one long-term extension study. Two cohorts were analyzed: a placebo-controlled cohort from 12- to 16-week studies and an all-abrocitinib cohort including patients who received one or more abrocitinib doses. Adverse events (AEs) of interest and laboratory data are reported. Total exposure in the all-abrocitinib cohort (n = 2856) was 1614 patient-years (PY); exposure was ≥ 24 weeks in 1248 patients and ≥ 48 weeks in 606 (maximum 108 weeks). In the placebo-controlled cohort (n = 1540), dose-related AEs (200 mg, 100 mg, placebo) were nausea (14.6%, 6.1%, 2.0%), headache (7.8%, 5.9%, 3.5%), and acne (4.7%, 1.6%, 0%). Platelet count was reduced transiently in a dose-dependent manner; 2/2718 patients (200-mg group) had confirmed platelet counts of < 50 × 103/mm3 at week 4. Incidence rates (IRs) were 2.33/100PY and 2.65/100 PY for serious infection, 4.34/100PY and 2.04/100PY for herpes zoster, and 11.83/100PY and 8.73/100PY for herpes simplex in the 200-mg and 100-mg groups, respectively. IRs for nonmelanoma skin cancer, other malignancies, and major adverse cardiovascular events were < 0.5/100PY for both doses. Five venous thromboembolism events occurred (IR 0.30/100PY), all in the 200-mg group. There were three deaths due to gastric carcinoma (diagnosed at day 43), sudden death, and COVID-19. Abrocitinib, with proper patient and dose selection, has a manageable tolerability and longer-term safety profile appropriate for long-term use in patients with moderate-to-severe AD. ClinicalTrials.gov: NCT02780167, NCT03349060, NCT03575871, NCT03720470, NCT03627767, NCT03422822.


18F-FDG PET/CT in Lymphomatoid Papulosis Mimicking Primary Cutaneous Anaplastic Large Cell Lymphoma.
Lymphomatoid papulosis is a benign self-healing condition, presenting as papulonodular skin eruptions and mimicking malignant cutaneous lymphomas histopathologically. F-FDG PET/CT findings in this benign condition have not been described in detail in the literature. We present a case of lymphomatoid papulosis mimicking primary cutaneous anaplastic large cell lymphoma histopathologically and demonstrating intensely FDG-avid cutaneous lesions on F-FDG PET/CT, which disappear spontaneously in the follow-up scan.


Writing Development in Associate Degree in Nursing-to-Baccalaureate Degree in Nursing Students: Moving Out of the Comfort Zone.
The nursing profession has made several unsuccessful attempts to differentiate practice between nurses with associate degrees and baccalaureate degrees entering the workforce. Writing competency has been identified as one potential area of differentiation. A descriptive, qualitative research design was used to explore Associate Degree in Nursing-to-Baccalaureate Degree in Nursing (ADN-to-BSN) graduates' perceptions of scholarly writing development in their baccalaureate studies. Nine recent graduates from a public college of nursing participated in semistructured interviews. Findings from content analysis revealed three phases of awareness in development of a scholarly writer: being a scholarly writer, assessing writing abilities, and connecting to practice. The study findings provide insight into competencies that could differentiate practice between ADN and BSN nurses entering the workforce. Although this study aimed to find out how ADN-to-BSN students are shaped as writers, findings suggest that the experience of learning to write may also be shaping students as professional nurses. [J Nurs Educ. 2017;56(3):182-185.].


Prognostic value of health status in patients with heart failure after acute myocardial infarction.
Disease-specific health status instruments such as the Kansas City Cardiomyopathy Questionnaire (KCCQ) can quantify symptoms, functional limitations, and quality of life in patients with heart failure. Understanding the relationship between KCCQ scores and prognosis may assist clinicians in both interpreting KCCQ scores and stratifying risk in patients. We examined the prognostic value of the KCCQ in a prospective, international cohort of 1516 patients with heart failure after a recent acute myocardial infarction. We focused on the relationship between the KCCQ overall score (KCCQ-os), measured at the first outpatient visit (4 weeks after enrollment), and subsequent 1-year cardiovascular mortality or hospitalization (n=258, 20.3%). KCCQ-os was strongly associated with subsequent cardiovascular events in that those with a score > or =75 had an 84% 1-year event-free survival compared with 59% for those with a score <25 (P<0.001). After demographic and other clinical characteristics were controlled for in multivariable models, KCCQ-os remained strongly associated with outcome (hazard ratio, 2.02; 95% CI, 1.24 to 3.27 for KCCQ-os <25; P<0.001). In outpatients with heart failure complicating an acute myocardial infarction, KCCQ-os is strongly associated with subsequent 1-year cardiovascular mortality and hospitalization. Use of the KCCQ in outpatient clinical practice can both quantify patients' health status and provide insight into their prognosis.


Similarity, connectionism, and the problem of representation in vision.
A representational scheme under which the ranking between represented similarities is isomorphic to the ranking between the corresponding shape similarities can support perfectly correct shape classification because it preserves the clustering of shapes according to the natural kinds prevailing in the external world. This article discusses the computational requirements of representation that preserves similarity ranks and points out the relative straightforwardness of its connectionist implementation.


Live imaging screen reveals that TYRO3 and GAK ensure accurate spindle positioning in human cells.
Proper spindle positioning is crucial for spatial cell division control. Spindle positioning in human cells relies on a ternary complex comprising Gαi1-3, LGN and NuMA, which anchors dynein at the cell cortex, thus enabling pulling forces to be exerted on astral microtubules. We develop a live imaging siRNA-based screen using stereotyped fibronectin micropatterns to uncover components modulating spindle positioning in human cells, testing 1280 genes, including all kinases and phosphatases. We thus discover 16 components whose inactivation dramatically perturbs spindle positioning, including tyrosine receptor kinase 3 (TYRO3) and cyclin G associated kinase (GAK). TYRO3 depletion results in excess NuMA and dynein at the cortex during metaphase, similar to the effect of blocking the TYRO3 downstream target phosphatidylinositol 3-kinase (PI3K). Furthermore, depletion of GAK leads to impaired astral microtubules, similar to the effect of downregulating the GAK-interactor Clathrin. Overall, our work uncovers components and mechanisms governing spindle positioning in human cells.


New Green Determination of Cu, Fe, Mn, and Zn in Beetroot Juices along with Their Chemical Fractionation by Solid-Phase Extraction.
A new simple and rapid method for the determination of the total concentrations of Cu, Fe, Mn, and Zn in beetroot juices by flame atomic absorption spectrometry was developed and validated. The method included a very simple sample preparation, i.e., the two-fold dilution and acidification of the samples with HNO3 to 1 mol·L-1 and provided the precision within 2%-3% and the trueness better than 6%. The method was applied for the rapid screening analysis of the different commercially available beetroot juices. The chemical fractionation of Cu, Fe, Mn, and Zn was also proposed by the two-column solid-phase extraction with the reversed-phase and cation exchange tubes. It was revealed that Cu, Fe, Mn, and Zn were primarily in beetroot juices in the form of the organically bound forms, contributing to the distinguished hydrophobic and residual fractions. The sums of the mean contributions of both fractions were up to 98% (Cu, Fe, Zn) and 100% (Mn), pointing out that no labile nor unbound forms of the studied metals were present in the matrix of beetroot juices.


A fully human antitumor immunoRNase selective for ErbB-2-positive carcinomas.
We report the preparation and characterization of a novel, fully human antitumor immunoRNase (IR). The IR, a human RNase and fusion protein made up of a human single chain variable fragment (scFv), is directed to the ErbB-2 receptor and overexpressed in many carcinomas. The anti-ErbB-2 IR, named hERB-hRNase, retains the enzymatic activity of the wild-type enzyme (human pancreatic RNase) and specifically binds to ErbB-2-positive cells with the high affinity (K(d) = 4.5 nm) of the parental scFv. hERB-hRNase behaves as an immunoprotoxin and on internalization by target cells becomes selectively cytotoxic in a dose-dependent manner at nanomolar concentrations. Administered in five doses of 1.5 mg/kg to mice bearing an ErbB-2-positive tumor, hERB-hRNase induced a dramatic reduction in tumor volume. hERB-hRNase is the first fully human antitumor IR produced thus far, with a high potential as a poorly immunogenic human drug devoid of nonspecific toxicity, directed against ErbB-2-positive malignancies.


[Subglottic granuloma with extubating mini-trach II: report of a case].
A case of subglottic granuloma after extubating Mini-trach II is described. The patient went aorto-coronary bypass and Mini-trach II was inserted for suction of sputum. Mini-trach II was extubated 6 days after insertion. The patient complained of dyspnea 3 months later. Examination showed subglottic granuloma. First, we burned the granuloma by laser under local anesthesia. Because the visual field was not good due to vocal cord movement, we failed to resect the granuloma completely. Therefore, we performed laryngo microsurgery under general anesthesia and resected the granuloma perfectly. The patient is in good health without signs of recurrence 6 months after granuloma removal. When we used Mini-trach II, the most important issues are insertion to the cricothyroid membrane, preventing infection, and careful follow-up after removing the Mini-trach II.


Acute thrombosis of a transplanted renal artery after gastric ulcer bleeding in a patient with a long-term well-functioning renal allograft: A case report and literature review.
Acute thrombosis of a transplanted renal artery is a serious vascular complication following renal allograft transplantation, which usually occurs within the first month after transplantation and often results in graft loss. It rarely occurs beyond the first month, except in a rejected kidney or in a kidney with high-grade transplant renal artery stenosis. A 65-year-old male with a history of type 2 diabetes mellitus, hypertension, pulmonary tuberculosis, and end-stage renal disease was previously treated with hemodialysis (HD). He received a kidney transplant and had a well-functioning graft for 2 years. He presented to our emergency department with gastric ulcer bleeding and received treatment involving an endoscopic submucosal epinephrine injection, a proton pump inhibitor, and blood transfusions. Nine days later, he complained of sudden lower abdominal pain and had acute anuric kidney failure. Renal ultrasonography revealed an absence of blood flow to the allograft kidney. Renal artery angiogram demonstrated complete occlusion of the transplanted renal artery. After thrombectomy and percutaneous transluminal angioplasty (PTA) with stent placement, 60% stenosis of the proximal renal artery with distal perfusion was noted. However, his graft function did not improve, and he received HD again. Histopathology of the transplanted kidney revealed ischemic tubular nephropathy with focal infarction without rejection. This is the first case of acute thrombosis of the transplanted renal artery following gastric ulcer bleeding in a patient with a long-term well-functioning graft kidney.


Calcium at the carotid siphon as an indicator of internal carotid artery stenosis.
Carotid siphon calcification is often visible on unenhanced head CT (UCT), but the relation to proximal carotid artery stenosis (CAS) is unclear. We investigated the association of carotid siphon calcification with the presence of CAS. This IRB-waived retrospective study included 160 consecutive patients suspected of stroke (age 64 ± 14 years, 63 female) who underwent head UCT and CTA of the head and neck. CAS was rated on CTA as not present or present with non-significant (<50 %), moderate (50-69 %) or significant (≥70 %) stenosis. Presence, shape (on UCT) and volume (on CTA) of carotid siphon calcifications were related to CAS. Carotid siphon calcification was absent in 41 % of patients and bilateral in 94 % of those with calcifications. Presence, shape and volume of calcification resulted in odds ratios for having significant CAS of 10.1, 3.9 and 8.4, with 95 % CIs of 1.3-79.6, 1.1-14.1 and 2.6-26.8, respectively. Corresponding NPVs were 0.98, 0.98 and 0.96, while PPVs were 0.14, 0.07 and 0.29, respectively. Absence of calcification in the carotid artery siphon on UCT has high negative predictive value for carotid artery stenosis in patients with suspected stroke. However, siphon calcification is not a reliable indicator of significant carotid artery stenosis. • Many stroke patients do not have calcification in the carotid artery siphon. • Carotid stenosis ≥50 % is unlikely in stroke patients without siphon calcification. • Carotid siphon calcium is a poor indicator of significant carotid artery stenosis.


[Identification of the genetic sex chromosomes in the monogenic blowfly Chrysomya rufifacies (Calliphoridae, Diptera)].
Previous investigations have shown the sex determination in the monogenic blowfly Chrysomya rufifacies to be controlled by a cytologically not discernible homogametry-heterogamety mechanism in the female. Female-producing (thelygenic) females are assumed to be heterozygous for a dominant female sex realizer (F') with sex-predetermining properties, while male-producing (arrhenogenic) females as well as males are supposed to be homozygous for the recessive allele (f). In order to identify the genetic sex chromosomes of C. rufifacies among its five pairs of long euchromatic chromosomes (nos. 1-5) plus one pair of small heterochromatic ones (no. 6), all chromosomes were marked by reciprocal translocations induced by X-ray treatment of adult males. The inheritance of thirteen different heteroxygous translocations has been analyzed. All of the translocations (eleven) between two of the four longer chromosomes did not show sex-linked inheritance, thus demonstrating the autosomal character of the chromosomes nos 1, 2, 3 and 4. The same is true for the translocation T6 (2/6). Therefore the small heterochromatic chromosome no. 6, corresponding to the morphlogically differentiated six chromosomes within the amphogenic calliphorid species, remains without sex determining function in the monogenic fly. This could be confirmed by the analysis of monosomic (monosomy-6) and trisomic (trisomy-6) individuals, which resulted from meiotic non-disfunction in T6/+ translocation heterozygotes. Contrary to these translocations, the heteroxygous 5/2 translocation (T14) exhibited sex-linked inheritance: There was but a very low frequency (0,76 per cent) of recombinants resulting from crossing-over between F'/f and the translocation breakage point in theylgenic F1 T14/+females. The sex-linked inheritance of T14 was confirmed by the progeny of a thelygenic F1 T14/+ female crossed to a homozygous T14/T14 translocation male.Among the offspring of that F1 T14/+ female, which had received the translocation from its father, all of the F2 T14/+ females were thelygenic compared to their arrhenogenic T14/T14 sisters. These results prove that the chromosomes of pair no. 5 genetically act as X'X-XX sex chromosomes in C. rufifacies.


Cancer treatment in the extreme elderly: case study of a 100-year-old lymphoma patient.
Limited data are available on the treatment of older adults with cancer. Comorbidities may preclude the administration of effective therapies, particularly in the extreme elderly. Comprehensive geriatric assessment can identify specific weaknesses of the patient and predict unexpected toxicities, thus enabling an optimized treatment strategy in this population. We report a case of the successful management of a 99-year-old female lymphoma patient with a strong wish for active treatment to improve quality of life and prolong survival past her 100th birthday. This case demonstrates that cancer treatment in the extreme elderly is possible and highlights the need for a formalized treatment plan based on geriatric assessment, frank discussion with patients and families, and defined goals of therapy.


Ghrelin counteracts salt-induced hypertension via promoting diuresis and renal nitric oxide production in Dahl rats.
Ghrelin is the endogenous ligand for the growth hormone-secretagogue receptor expressed in various tissues including the heart, blood vessels and kidney. This study sought to determine the effects of long-term treatment with ghrelin (10 nmol/kg, twice a day, intraperitoneally) on the hypertension induced by high salt (8.0% NaCl) diet in Dahl salt-sensitive hypertensive (DS) rats. Systolic blood pressure (SBP) was measured by a tail cuff method. During the treatment period for 3 weeks, high salt diet increased blood pressure compared to normal salt (0.3% NaCl) diet, and this hypertension was partly but significantly (P<0.01) attenuated by simultaneous treatment with ghrelin. Ghrelin significantly increased urine volume and tended to increase urine Na⁺ excretion. Furthermore, ghrelin increased urine nitric oxide (NO) excretion and tended to increase renal neuronal nitric oxide synthase (nNOS) mRNA expression. Ghrelin did not alter the plasma angiotensin II level and renin activity, nor urine catecholamine levels. Furthermore, ghrelin prevented the high salt-induced increases in heart thickness and plasma ANP mRNA expression. These results demonstrate that long-term ghrelin treatment counteracts salt-induced hypertension in DS rats primarily through diuretic action associated with increased renal NO production, thereby exerting cardio-protective effects.


Comminuted epiphyseal fracture of the distal tibia. A case report and review of the literature.
Classification of epiphyseal injuries include only simple fractures and few cases of comminuted fracture of the distal tibial epiphysis, epiphyseal plate and adjacent metaphysis. On the basis of a review of the literature, as well as our own experience, it appears that that fracture is unstable and requires open reduction and internal fixation.


Histopathologic reaction of a calcium phosphate cement for alveolar ridge augmentation.
The objective of the present study was to evaluate the feasibility of using a calcium phosphate cement (CPC) in the reconstruction of a defective alveolar ridge in conjunction with implant placement. The CPC consisted of an equimolar amount of tetracalcium phosphate and dicalcium phosphate anhydrous. At the beginning of the experiment, all mandibular premolar teeth of mature beagle dogs were extracted. After 1 month of healing, alveolar bone was reduced to make a space for a CPC block that was prefabricated from a CPC mixed with water at a powder/liquid ratio of 5 g/mL. After an additional month, 8-mm long hydroxyapatite-coated titanium implants were placed in such a way that the apical half was embedded into alveolar bone and the coronal half in the preformed CPC block. The dogs were sacrificed and biopsies were obtained at 1, 3, and 6 months after surgery. Sections that included implants were evaluated for integration of the CPC block to the alveolar bone and of the implant to the alveolar bone. Additional sections without the implants served as controls. The results obtained from this study show that the CPC ridge augmentation gradually is replaced by natural bone. Six months after surgery, histopathologic features of the augmentation area were quite similar to those of natural alveolar bone. The coronal half of the implants, previously surrounded by the CPC block, was firmly fixed by natural bone. Therefore, this method may be useful for increasing the height of the alveolar ridge.


[pH-dependence of the esterolytic activity of terrylytine proteases].
The pH-dependences of the kinetic parameters of hydrolysis of alpha-N-henzoyl-L-arginine and N-benzoyl-L-tyrosine ethyl esters by terrylytine proteases I and II were studied. The experimental results proved evidence for the differences in the activities of these enzymes. The active center of protease I contains two groups with close pK values (6,5-7,0), one of which is presumably the imidazole group of histidine. The catalytic action of protease II is determined by the state of the ionizing groups with the ionization constants of about 6 and 8. The former functions in the deprotonated form, whereas the latter--in the protonated form.


Transcatheter aortic valve replacement using the new Evolut-Pro system: a prospective comparison with the Evolut-R device.
Evolut Pro (EVP) is a novel self-expandable aortic valve. This prosthesis consists of an external porcine pericardial wrap designed to reduce paravalvular leak (PVL), maintaining the benefits of its predecessor, the Evolut R (EVR). The aim was to compare the functional and clinical results in the short and medium term of the new EVP with the EVR system. Consecutive patients receiving either the EVR (n=50) or the EVP (n=33) from June 2015 to October 2018 were compared. Baseline characteristics, cardiovascular imaging, procedural outcomes, short and mid-term follow-up outcomes were prospectively collected and assessed. Residual mild PVL was common and comparable in the two groups (EVR 79% vs. EVP 70%; P=0.4). In the EVR group, the presence of PVL was directly related to prosthesis size, but this correlation was not observed in the EVP group. Conduction abnormalities were more prevalent with the EVP, but these did not translate into a higher need of permanent pacemaker implantation. Vascular and bleeding complications were infrequent in both groups. At mid-term clinical follow-up (median survival time: EVR 11±0.3 months, EVP 12±0.2 months), the 1-year rate of adverse events was similar (EVR: 24%, EVP: 33%; P=0.3). Both protheses are effective for the treatment of severe aortic stenosis with excellent results at mid-term clinical follow up. The EVP remains associated with a significant rate of residual mild PVL that appears to be similar to that observed with EVR.


Highlights for α-fetoprotein in determining prognosis and treatment monitoring for hepatocellular carcinoma.
To explore the prognostic value in the monitoring of treatment efficacy of serial α-fetoprotein (AFP) in hepatocellular carcinoma (HCC) patients. We searched MEDLINE, EMBASE and COCHRANE LIBRARY through April 21, 2012, to find qualifying articles. Our overall search strategy included terms for HCC, AFP, treatment response, and prognosis. Literature was limited to English-language, human studies. Studies reporting cumulative survival rates were summarized qualitatively. For the prognostic meta-analysis, we undertook a series of meta-analyses that summarised the Cox proportional hazard ratios (HRs) by assuming a random effects model. With regards to the correlation of AFP change with radiologic response, the categorical dichotomous variables were assessed using Poisson relative risks (RRs), which were incorporated into the random effects model meta-analysis of accuracy prediction. Between-study heterogeneity was estimated by use of the I² statistic. Publication bias was evaluated using the Begg funnel plot and Egger plot. Sensitivity analyses were conducted first by separating systemic treatment estimates from locoregional therapy estimates, evaluating different AFP response cut-off point effects, and exploring the impact of different study sizes. Of 142 titles identified in our original search, 11 articles (12 clinical studies) met our criteria. Six studies investigated outcome in a total of 464 cases who underwent systemic treatment, and six studies investigated outcome in a total of 510 patients who received locoregional therapy. A random-effects model meta-analysis showed that AFP response was associated with an mortality HR of 0.55 (95%CI, 0.47-0.65) across HCC in overall survival (OS) and 0.50 (95%CI, 0.38-0.65) in progression-free survival. Restricting analysis to the six eligible analyses of systemic treatment, the pooled HRs were 0.64 (95%CI, 0.53-0.77) for OS. Limiting analysis to the six analyses of locoregional therapy, the pooled HRs for OS was 0.39 (95%CI, 0.29-0.53). We showed a larger pooled HR in the 50% definition studies (HR, 0.67, 95%CI, 0.55-0.83) compared with that from the 20% definition studies (HR, 0.41, 95%CI, 0.32-0.53). Restricting analysis to the four studies including over 100 patients individually, the pooled HR was 0.65 (95%CI, 0.54-0.79), with a pooled HR for OS of 0.35 (95%CI, 0.23-0.46) in the studies of less than 100 patients. As to radiological imaging, 43.1% (155/360) of the patients in the AFP response group presented with a radiological overall response, while the response rate decreased to 11.5% (36/313) in the patients from the AFP nonresponse group. The RR of having no overall response was significantly lower in the AFP response group than the AFP nonresponse group (RR, 0.67; 95%CI, 0.61-0.75). In terms of disease control rate, 86.9% (287/330) in the AFP response group and 51.0% (153/300) in the AFP nonresponse group showed successful disease control, respectively. The RR of disease control failure, similarly, was significantly lower in the AFP response group (RR, 0.37; 95%CI, 0.23-0.58). But these findings could be overestimates because of publication and reporting bias. HCC patients presenting with an AFP response are at decreased risk of mortality. In addition, patients with an AFP response also present with a higher overall response rate and disease control rate.


Natural products modulate NLRP3 in ulcerative colitis.
Ulcerative colitis (UC) is a clinically common, progressive, devastating, chronic inflammatory disease of the intestine that is recurrent and difficult to treat. Nod-like receptor protein 3 (NLRP3) is a protein complex composed of multiple proteins whose formation activates cysteine aspartate protease-1 (caspase-1) to induce the maturation and secretion of inflammatory mediators such as interleukin (IL)-1β and IL-18, promoting the development of inflammatory responses. Recent studies have shown that NLRP3 is associated with UC susceptibility, and that it maintains a stable intestinal environment by responding to a wide range of pathogenic microorganisms. The mainstay of treatment for UC is to control inflammation and relieve symptoms. Despite a certain curative effect, there are problems such as easy recurrence after drug withdrawal and many side effects associated with long-term medication. NLRP3 serves as a core link in the inflammatory response. If the relationship between NLRP3 and gut microbes and inflammation-associated factors can be analyzed concerning its related inflammatory signaling pathways, its expression status as well as specific mechanism in the course of IBD can be elucidated and further considered for clinical diagnosis and treatment of IBD, it is expected that the development of lead compounds targeting the NLRP3 inflammasome can be developed for the treatment of IBD. Research into the prevention and treatment of UC, which has become a hotbed of research in recent years, has shown that natural products are rich in therapeutic means, and multi-targets, with fewer adverse effects. Natural products have shown promise in treating UC in numerous basic and clinical trials over the past few years. This paper describes the regulatory role of the NLRP3 inflammasome in UC and the mechanism of recent natural products targeting NLRP3 against UC, which provides a reference for the clinical treatment of this disease.


Mass Spectrometry Using Nanomechanical Systems: Beyond the Point-Mass Approximation.
The mass measurement of single molecules, in real time, is performed routinely using resonant nanomechanical devices. This approach models the molecules as point particles. A recent development now allows the spatial extent (and, indeed, image) of the adsorbate to be characterized using multimode measurements ( Hanay , M. S. , Nature Nanotechnol. , 10 , 2015 , pp 339 - 344 ). This "inertial imaging" capability is achieved through virtual re-engineering of the resonator's vibrating modes, by linear superposition of their measured frequency shifts. Here, we present a complementary and simplified methodology for the analysis of these inertial imaging measurements that exhibits similar performance while streamlining implementation. This development, together with the software that we provide, enables the broad implementation of inertial imaging that opens the door to a range of novel characterization studies of nanoscale adsorbates.


Characterization of the effects of 2-methylthio-ATP and 2-chloro-ATP on brain capillary endothelial cells: similarities to ADP and differences from ATP.
1. Brain capillary endothelial cells responded to 2-methylthio-ATP (2MeSATP) by large increases in [Ca2+]i (EC50 = 27 nM) that were partially dependent on the presence of extracellular Ca2+ and that were not associated with a measurable production of inositol phosphates. 2. 2-chloro-ATP (2ClATP) raised [Ca2+]i in a biphasic manner. At low concentrations, intracellular Ca2+ mobilization was not associated with a measurable production of inositol phosphates. At concentrations > 30 microM, 2ClATP activated phospholipase C. 3. The actions of 2ClATP, 2MeSATP and ADP on [Ca2+]i were additive to those of ATP and UTP. Non-additive actions of 2MeSATP and of low concentrations of ADP or of 2ClATP were observed. 4. Cross desensitizations of the actions of ADP, 2MeSATP and 2ClATP were observed. None of them desensitized cells to the action of ATP. 5. It is concluded that 2MeSATP and low concentrations of 2ClATP and ADP induce intracellular Ca2+ mobilization by acting via an atypical P2y purinoceptor that is not coupled to phospholipase C. At high concentrations, 2ClATP also activates phospholipase C and further increases [Ca2+]i probably by acting on P2u purinoceptors.


Investigation of anti-Toxoplasma gondii antibodies in cats of the Ankara region of Turkey Using the Sabin-Feldman dye test and an indirect fluorescent antibody test.
Blood samples from 99 cats from the Ankara province of Turkey were examined for the presence of anti-Toxoplasma gondii antibody with the use of both the Sabin-Feldman dye test (DT) and an indirect fluorescent antibody test (IFAT). Forty of the 99 sera (40.3%) were positive for antibodies against T. gondii with the DT, whereas the IFAT assay detected antibodies in 34 (34.3%). The study also evaluated 3 factors for their potential association with the presence of T. gondii antibody: age (<1 yr, 1-2 yr, and >2 yr), gender (female vs. male), and outdoor access (stray, owned with outdoor access, or indoor only). The DT detected antibodies in 3 cats under 1 yr of age, 22 cats between 1 and 2 yr, and 15 cats older than 2 yr, whereas the IFAT found 1, 18, and 15 cats positive for antibodies, respectively, in each of these categories. Of 61 female cats, 27 (44.2%) were positive by the DT; and of 38 male cats, 13 (34%) were positive by the DT. For the IFAT, 24 female cats (39.3%) and 10 male cats (26.3%) were positive. The percent seropositivity in indoor cats was 30.8% by the DT and 23.1% by the IFAT. In stray cats, the percent seropositivity was 52.8% by the DT and 41.7% by the IFAT. Antibody presence was significantly associated with age, but not with outdoor access.


Non-invasive, multimodal analysis of cortical activity, blood volume and neurovascular coupling in infantile spasms using EEG-fNIRS monitoring.
Although infantile spasms can be caused by a variety of etiologies, the clinical features are stereotypical. The neuronal and vascular mechanisms that contribute to the emergence of infantile spasms are not well understood. We performed a multimodal study by simultaneously recording electroencephalogram and functional Near-infrared spectroscopy in an intentionally heterogeneous population of six children with spasms in clusters. Regardless of the etiology, spasms were accompanied by two phases of hemodynamic changes; an initial change in the cerebral blood volume (simultaneously with each spasm) followed by a neurovascular coupling in all children except for the one with a large porencephalic cyst. Changes in cerebral blood volume, like the neurovascular coupling, occurred over frontal areas in all patients regardless of any brain damage suggesting a diffuse hemodynamic cortical response. The simultaneous motor activation and changes in cerebral blood volume might result from the involvement of the brainstem. The inconstant neurovascular coupling phase suggests a diffuse activation of the brain likely resulting too from the brainstem involvement that might trigger diffuse changes in cortical excitability.


On the physical basis of asymmetry and homochirality.
Mirror symmetry breaking is ubiquitous in our visible universe taking place in elementary particles, atoms, and molecules. Molecular chirality is not biogenic in itself, although its detection is often considered a biosignature, a conjecture inferred from the fact that we do not know life devoid of homochirality. The question of whether there is a connection between the cosmic preference for one enantiomer, as imposed by the weak force, and the single chirality displayed on Earth is vividly debated. This article gives a glimpse on the origin of asymmetry from a cosmological perspective and on physical transformations that lead to an enantiomeric imbalance, leaving chemical reactions essentially aside. These processes are more plausible as sources of prebiotic chirality than asymmetric amplifications requiring unnatural substrates and conditions and fighting against racemization. The latter may actually be a friend, not foe, and a driving force for enantioselection.


Anti-Interleukin-9 Antibody Increases the Effect of Allergen-Specific Immunotherapy in Murine Allergic Rhinitis.
Interleukin (IL)-9 induces allergic responses; however, the roles of anti-IL-9 antibody in the induction of tolerance remain unclear. This study investigated the effects of anti-IL-9 antibody on oral tolerance (OT) in a mouse model of allergic rhinitis (AR). BALB/c mice were divided into 4 groups: the control, AR, OT, and OT with anti-IL-9 antibody (OT+IL9AB) groups. Ovalbumin (OVA) was used for sensitization and challenge. Mice in the OT and OT+IL9AB groups were fed OVA for immunotherapy. During immunotherapy, OT+IL9AB mice were injected with anti-IL-9 antibody. Allergic symptoms, tissue eosinophil counts, and serum OVA-specific immunoglobulin E (IgE) were measured. The mRNA expressions of cytokines and transcription factors of T cells of nasal mucosa were determined by real-time polymerase chain reaction (PCR). The protein levels of GATA3, ROR-γt, and Foxp3 in nasal mucosa were determined by Western blot. CD4⁺CD25⁺Foxp3⁺ T cells in the spleen were analyzed by flow cytometry. Administration of anti-IL-9 antibody decreased allergic symptoms, OVA-specific IgE levels, and eosinophil counts. In addition, it inhibited T-helper (Th) 2 responses, but had no effect on Th1 responses. Protein levels of ROR-γt and mRNA levels of PU.1 and ROR-γt were reduced by anti-IL-9 antibody. Anti-IL-9 antibody increased Foxp3 and IL-10 mRNA expression, Foxp3 protein, and induction of CD4⁺CD25⁺Foxp3⁺ T cells. Anti-IL-9 antibody decreased allergic inflammation through suppression of Th2 and Th17 cells. Anti-IL-9 antibody enhanced the tolerogenic effects of regulatory T cells. These results suggest that anti-IL-9 antibody might represent a potential therapeutic agent for allergen immunotherapy in patients with uncontrolled allergic airway disease.


Hospital-based event medical support for the Baltimore Marathon, 2002-2005.
We present a four-year, cross-sectional epidemiologic description of injuries and illnesses among Baltimore Marathon participants and the evaluation, treatment, and disposition of those conditions by an on-site event medical team led by physicians and staff from an urban, academic emergency department. We analyzed data from injuries encountered during the marathon. Subjects presenting to a medical aid station along the course route or at the finish line were defined as "injured or ill" and were prospectively divided into two groups: 1) a brief-encounter group and 2) an extended-encounter group. Data collected included gender, presenting complaint(s), assessment, treatment(s), and disposition. Three percent (N=1,144) of approximately 33,700 total participants over four years presented to medical aid stations during the Baltimore Marathon between 2002 and 2005. Most participants (66%) did not require a full clinical evaluation. Common complaints encountered were dehydration (32%), musculoskeletal injuries (25%), and cutaneous wounds (20%). Transport to the hospital was required for 4% of all injured participants, and 61% returned to the race. Most injuries/illnesses encountered at the Baltimore Marathon in 2002-2005 were minor, although some were serious enough to require transport to a hospital. The year with the highest average race-day temperature had the highest observed injury rates and the highest number of hospital transports. These results help to improve understanding of the types, severity, and distribution of injuries commonly sustained by marathon participants and may guide decisions regarding the appropriate distribution of emergency medical resources at such events.


Distinguishing steric and electrostatic molecular probe orientational ordering via their effects on reorientation-induced spectral diffusion.
The theoretical framework for reorientation-induced spectral diffusion (RISD) describes the polarization dependence of spectral diffusion dynamics as measured with two-dimensional (2D) correlation spectroscopy and related techniques. Generally, RISD relates to the orientational dynamics of the molecular chromophore relative to local electric fields of the medium. The predictions of RISD have been shown to be very sensitive to both restricted orientational dynamics (generally arising from steric hindrance) and the distribution of local electric fields relative to the probe (electrostatic ordering). Here, a theory that combines the two effects is developed analytically and supported with numerical calculations. The combined effects can smoothly vary the polarization dependence of spectral diffusion from the purely steric case (least polarization dependence) to the purely electrostatic case (greatest polarization dependence). Analytic approximations of the modified RISD equations were also developed using the orientational dynamics of the molecular probe and two order parameters describing the degree of electrostatic ordering. It was found that frequency-dependent orientational dynamics are a possible consequence of the combined electrostatic and steric effects, providing a test for the applicability of this model to experimental systems. The modified RISD equations were then used to successfully describe the anomalous polarization-dependent spectral diffusion seen in 2D infrared spectroscopy in a polystyrene oligomer system that exhibits frequency-dependent orientational dynamics. The degree of polarization-dependent spectral diffusion enables the extent of electrostatic ordering in a chemical system to be quantified and distinguished from steric ordering.


Oncolytic Virotherapy and Gene Therapy Strategies for Hepatobiliary Cancers.
Advanced liver cancers and biliary cancers represent diseases with dismal prognosis because of frequent local invasion and metastasis. Effective therapeutic agents for these cancers have not been established. Oncolytic viruses (OVs) constitute a novel class of promising, selective anticancer agents and recent studies have elucidated their unique features. Moreover, clinical trials are demonstrating promising results. Numerous OVs are being tested in preclinical models of hepatocellular carcinoma (HCC). The lead agent Pexa-Vec (pexastimogene devacirepvec, JX-594), a recombinant Wyeth strain vaccinia virus, has demonstrated preliminary evidence of safety and efficacy for HCC in clinical trials. Few other OVs have entered clinical testing. Relatively few preclinical studies and clinical trials exist for biliary cancers. In this review, we introduce various approaches using OVs to treat the intractable hepatobiliary cancers.


Sodium Intake Among Persons Aged ≥2 Years - United States, 2013-2014.
High sodium consumption can increase hypertension, a major risk factor for cardiovascular diseases (1). Reducing sodium intake can lower blood pressure, and sodium reduction in the U.S. population of 40% over 10 years might save at least 280,000 lives (2). Average sodium intake in the United States remains in excess of Healthy People 2020 objectives,* and monitoring sources of sodium in the U.S. population can help focus sodium reduction measures (3,4). Data from 2013-2014 What We Eat in America (WWEIA), the dietary intake portion of the National Health and Nutrition Examination Survey (NHANES),† were analyzed to determine the ranked percentage sodium contribution of selected food categories and sources of sodium intake from all reported foods and beverages, both overall and by demographic subgroups. These latest data include updated food codes and separate estimates for intake among non-Hispanic Asians.§ In 2013-2014, 70% of dietary sodium consumed by persons in the United States came from 25 food categories; breads were the top contributor, accounting for 6% of sodium consumed. A majority of sodium consumed was from food obtained at stores; however, sodium density (mg/1,000 kcal) was highest in food obtained at restaurants. A variety of commonly consumed foods contributes to U.S. sodium intake, emphasizing the importance of sodium reduction across the food supply (4).


Pedicled vascularized bone grafts for disorders of the carpus: scaphoid nonunion and Kienbock's disease.
The use of reverse-flow pedicled vascularized bone grafts from the dorsal distal radius makes it possible to transfer bone with a preserved circulation and viable osteoclasts and osteoblasts. The resultant primary bone healing without creeping substitution within the dead bone is an alternative to conventional bone grafting in aiding or accelerating healing, replacing deficient bone, and/or revascularizing ischemic bone. Recent advances in understanding the anatomy and physiology of vascularized pedicled bone grafts have increased their use in treating a variety of carpal maladies. A basic understanding of the vascular anatomy, as well as the surgical principles and experimental and clinical results of pedicled vascularized bone grafts from the dorsal distal radius, is critical to understanding the use of these grafts in the treatment of scaphoid nonunions and Kienbock's disease.


An Introduction to the Inverted/Flipped Classroom Model in Education and Advanced Training in Medicine and in the Healthcare Professions.
In describing the inverted classroom model (ICM), the following paper is meant to provide an introduction to the subject matter and to serve as a practical guide for those wishing to employ its methods in basic and advanced medical training and education. The ICM is a blended-learning method in which a self-directed learning phase (individual phase) precedes the classroom-instruction phase. During the online phase, factual knowledge is imparted that serves as a basis for the classroom phase. The classroom phase should subsequently be used to assimilate and implement the previously gained knowledge. In contrast, traditional course concepts impart factual knowledge in lectures, for example, or in other face-to-face teaching formats and are followed by the students' self-instruction in order to assimilate this knowledge. The goal of the ICM is the shift from passive learning to accelerated learning in order to foster learning at cognitively demanding levels such as analysis, synthesis and evaluation. The concurrent increase in production and use of screencasts and educational videos, the Open Educational Resources "movement" and the widespread use of Massive Open Online Courses (MOOCS) have contributed to the increased dissemination of the inverted-classroom method. The intention of the present paper is to provide an introduction to the subject matter and simultaneously to offer a short overview of important projects and research results in the field of medical education and other health professions. Furthermore, an outline is given of the advantages and disadvantages of the model as well as its potential benefit to the future of medical education and training.


Role of bone marrow trephine biopsy in the diagnosis of marrow involvement in Hodgkin's disease.
Bone marrow involvement in Hodgkin's disease alters the stage and hence the prognosis and treatment modality. Seventy-six patients of biopsy proven Hodgkin's disease in whom a bone marrow aspiration and biopsy were done simultaneously at the time of diagnosis were included in the study. Bone marrow involvement was seen in 14 (18%) cases. All 14 cases were diagnosed on biopsy. Only one aspirate was suspicious of involvement. Rest was either diluted (28%) or was normal (64%). Eleven patients were upstaged from stage I, II and III to stage IV. All patients had cytopenia of one or more cell line. Increased fibrosis, necrosis and a polymorphic cellular infiltrate were associated findings in the marrow. We wish to highlight the importance of bone marrow trephine biopsy over aspirate in staging even clinically early stage Hodgkin's disease.


Torsade de pointes after amiodarone withdrawal; effects of mild hypokalaemia on repolarization.
Recurrent atypical ventricular tachycardia, with a long QT interval, was documented in a 56-year-old patient 16 days after discontinuing amiodarone 200 mg and 400 mg on alternate days. Tachycardia and variable prolongation of myocardial repolarization were abolished by temporary cardiac pacing and correction of mild hypokalaemia.


The prevalence and predictors of emotional distress and social difficulties among surviving cancer patients in Jordan.
This study examines the prevalence and degree of emotional distress, anxiety and depression, and social difficulties and their effect on cancer patients' quality of life (QoL). It describes the characteristics of patients who are at high risk of emotional distress. A descriptive cross-sectional design was used. A total of 226 patients with cancer completed the Hospital Anxiety and Depression scale (HAD), social difficulties inventory, comfort scale and EORTC-QoL-C30. Anxiety and depression were identified using the internationally recognized cut-off points of HAD-A ≥ 8 and HAD-D≥ 8. Adjusted odd ratio was calculated using socio-demographic and clinical factors. Both anxiety and depression were common among Jordanian cancer patients, although depression was the main emotional problem with a higher prevalence than anxiety (67.6% vs. 43%). Patients with anxiety or depression were more likely to have lower QoL scores and higher scores for complaints about symptoms. They were more likely to have social difficulties in everyday life. The results of logistic regression indicated that a high depression score was predicted by older age, a poor QOL total score, and a high social difficulty score. A high anxiety score was predicted by advanced cancer stage, in female patients, hospital readmission and a poor QOL total score. The significant level of emotional distress among cancer patients highlighted the importance of early assessment and identification of patients at greater risk of emotional distress, those with an advanced stage of cancer, having a poor quality of life and serious social difficulties.


Restoring meaning in life by meaning-focused coping: The role of self-distancing.
Meaning-focused coping is an important coping strategy in the face of stressful life events. The "construal level theory" and recent research suggest that reflecting on a stressful situation from a self-distanced stance (temporal or spatial) allows individuals to find meaning in negative life events; however, its long-term effects have not yet been explored. The present research had two main goals: (1) to determine whether distanced meaning making enhances meaning in life and (2) to explore the underlying emotional mechanism of this effect. We hypothesized that positive affect (relative to negative affect) may play a more important role in the relationship between self-distanced meaning making and meaning in life. A total of 136 university students were recruited as participants in two studies. The participants were prospectively instructed to make meaning of a highly stressful event. They were randomly assigned to one of three conditions: (a) self-immersed, (b) temporally distanced, or (c) spatially distanced. The findings indicate that both the temporally and spatially distanced perspectives in the meaning-making process can enhance meaning in life and that they do so by promoting positive affect. Implications for future research on meaning-focused coping and self-distancing are discussed.


The yeast aminopeptidase Y.
A metal-dependent aminopeptidase (EC 3.4.11.-), designated APase Y, has been purified to homogeneity by conventional methods. The enzyme is composed of a single polypeptide chain with molecular mass of 102 kilodaltons, estimated by sodium dodecyl sulphate - polyacrylamide gel electrophoresis, with a blocked N-terminal amino acid. It possesses neither endopeptidase nor carboxypeptidase activity and is strongly inhibited by metal-chelating agents, Zn2+, and the protein inhibitor from Neurospora crassa. APase Y is insensitive to Cl anions, S--S reducing reagents, serine protease inhibitors, and the peptidase inhibitor benzamidine. Co2+, Hg2+, and p-chloromercuribenzoate can activate the enzyme up to 22, 20, and 55%, respectively. The holoenzyme is resistant to yeast endopeptidases A, B, and Y, whereas the apoenzyme (obtained after treatment with chelators) is susceptible to the serine endopeptidases B and Y. The enzyme catalyzes hydrolysis of most L peptides possessing free alpha-amino (or imino) group by stepwise removal of N-terminal residue. Peptides with L-leucine at the N terminus are cleaved preferentially. The enzyme is unable to catalyze hydrolysis of X--Pro type peptide bonds, and inefficiently hydrolyzes bonds between Asp--X and Glu--X. L-leucine p-nitroanilide hydrolyzes optimally at pH 8.2 with a Km value of 1 mM. The purified enzyme is stable during storage in 0.05 M phosphate buffer, pH 6.7, containing 40-50% glycerol, at -20 degrees C.


Modular fluorescent-labeled siderophore analogues.
Biomimetic analogues 1 of the microbial siderophore (iron carrier) ferrichrome were labeled via piperazine with various fluorescent markers at a site not interfering with iron binding or receptor recognition (compounds 10-12). These iron carriers were built from a tetrahedral carbon symmetrically extended with three strands, each containing an amino acid (G = glycyl, A = alanyl, L = leucyl and P = phenylalanyl) and terminated by a hydroxamic acid, which together define an octahedral iron-binding domain. A fourth exogenous strand provided the site for connecting various fluorescent markers via a short bifunctional linker. Iron(III) titrations, along with fluorescence spectroscopy, generated quenching of fluorescence emission of some of the probes used. The quenching process fits the Perrin model which reinforces the intramolecular quenching process, postulated previously.1 All tested compounds, regardless of their probe size, polarity, or the linker binding them to the siderophore analogue, promote growth of Pseudomonas putida with the same efficacy as the nonlabeled analogues 1, with the added benefit of signaling microbial activity by fluorescence emission. All G derivatives of compounds 10-12 were found to parallel the behavior of natural ferrichrome, whereas A derivatives mediated only a modest iron(III) uptake by P. putida. Incubation of various Pseudomonas strains with iron(III)-loaded G derivatives resulted in the build-up of the labels' fluorescence in the culture medium to a much larger extent than from the corresponding A derivatives. The fluorescence buildup corresponds to iron utilization by the cells and the release of the fluorescent labeled desferrisiderophore from the cell to the media. The fact that the microbial activity of these compounds is not altered by attachment of various fluorescent markers via a bifunctional linker proposes their application as diagnostic tools for detecting and identifying pathogenic microorganisms.


The Group B Streptococcus NADH oxidase Nox-2 is involved in fatty acid biosynthesis during aerobic growth and contributes to virulence.
Numerous Streptococcaceae produce an H2O-forming NADH oxidase, Nox-2, which has been generally implicated in aerobic survival. We examined the roles of Nox-2 in Group B Streptococcus (GBS), a leading agent of neonatal infections. While nox2 inactivation caused an aerobic growth arrest, no improvement was seen by addition of antioxidants to cultures, suggesting that this defect was not due to accumulation of toxic oxygen species. Using several approaches, we show that the observed inability of the nox2 mutant to grow aerobically is mainly due to an underlying defect in fatty acid (FA) biosynthesis: (i) the nox2 aerobic growth defect is fully and rapidly complemented by adding oleic acid to culture medium, and (ii) direct assimilation of this unsaturated FA in both wild type (WT) and nox2 GBS membranes is demonstrated and correlated with mutant growth rescue. We propose that NAD+ depletion in the nox2 mutant results in reduced acetyl-CoA production, which perturbs FA biosynthesis and hence blocks growth in aerobiosis. The nox2 aerobic growth defect was also complemented when GBS respiration metabolism was activated by exogenous haem and menaquinone. The membrane NADH oxidase activity generated by the functional respiratory chain thus compensates the cytoplasmic NADH oxidase deficiency. The nox2 mutant was attenuated for virulence, as assessed in lung, intraperitoneal and intravenous murine infection models. As the nox2 defect seems only to affect aerobic growth of GBS, its reduced virulence supports the suggestion that aerobic conditions and NADH oxidase activities are relevant to the GBS infection process.


Knocking-down of CREPT prohibits the progression of oral squamous cell carcinoma and suppresses cyclin D1 and c-Myc expression.
As a regulator essential for many cell cycle-related proteins, the robust expression of Cell cycle-Related and Expression-elevated Protein in Tumor (CREPT) implicates a poor diagnosis of endoderm and mesoderm-derived tumors. Whether CREPT plays the same role in the tumorigenesis derived from ectodermal tissues remains elusive. To explore the role of CREPT in ectoderm-derived tumors, cells from 7oral squamous cell carcinoma (OSCC) lines and 84clinical OSCC samples were exploited in this study. Quantitative PCR, Western blot assay and immunohistochemistry were applied in the evaluation of CREPT, cyclin D1 and c-Myc expression. Knocking-down of CREPT was performed by lentivirus delivering specific shRNA of CREPT. The effects of CREPT on OSCC were examined by cell proliferation, colony formation, apoptosis, cell migration and xenograft implantation experiments. Compared with human normal oral keratinocytes, OSCC cell lines showed a significantly elevated expression of CREPT in both mRNA and protein levels. Consistently, samples from OSCC patients also exhibited a noticeably stronger CREPT expression than the noncancerous samples. In contrast, knocking down of CREPT in OSCC cell lines significantly reduced proliferation, colony formation and migration as well as the expression of cyclin D1 and c-Myc, but promoted apoptosis. Statistical analysis also suggested that CREPT expression was significantly correlated with the T and N classification of OSCC. Furthermore, CAL27 mouse xenograft model confirmed that down-regulation of CREPT prohibited cyclin D1 and c-Myc expression, through which decreased the in vivo tumor growth, but increased the survival ratio of hosts. In OSCC cell lines, up-regulated CREPT expression enhanced cell proliferation, migration and cell cycle as well as promoted cyclin D1 and c-Myc expression as it did in endoderm and mesoderm-origin tumors. Our study strongly suggests that CREPT could be used as a marker for the OSCC prognosis and might work as a potential target in future OSCC therapy.


Alternative splicing generates a novel non-secretable cytosolic isoform of NELL2.
NELL2 is as a neuron-specific secreted glycoprotein. The present study provides evidence of an alternatively spliced variant of the rat NELL2 gene that yields cytosolic NELL2 (cNELL2). cNELL2 was initially detected in the thymus and subsequently found to be ubiquitously expressed in many other tissues. The absence of the sequences corresponding to the third exon, which contains the terminal portion of the signal peptide, accounts for the uniform distribution of cNELL2 throughout the cytoplasm. This is in contrast to NELL2, which is preferentially located at distinct subcellular structures involved in the secretary process, such as endoplasmic reticulum and Golgi apparatus. Western blot analysis showed that cNELL2 was not present in the medium but only in lysates, while NELL2 was detected as a glycosylated larger form in both lysates and media. Immunoprecipitation analysis revealed that cNELL2 interacts with PKCbeta1. These results suggest that cNELL2 is involved in PKCbeta1-mediated intracellular signaling.