Effect of activated carbon microstructure and adsorption mechanism on the efficient removal of chlorophyll a and chlorophyll b from Andrographis paniculata extract.
In order to reveal the effect of activated carbon (AC) properties on the adsorption of chlorophyll a (Chl a) and chlorophyll b (Chl b) in Andrographis paniculata extract, four commercial activated carbons were first tested and characterized. The results showed that activated carbon 1 (AC1) had the best surface area, pore structure and adsorption capacity. Therefore, adsorption isotherms, adsorption kinetics and adsorption mechanism were further carried out on AC1. The application of Langmuir model (R2 > 0.978) and Freundlich model (R2 > 0.977) indicated that the adsorption process of Chl a and Chl b on AC1 may be a complex adsorption process of single-layer and multilayer adsorption. The adsorption kinetics indicated that the pseudo-second-order kinetic model (R2 > 0.999) was dominant and was mainly chemisorption. The intra-particle diffusion model (R2 > 0.937) shows that the intra-particle diffusion is the rate-limiting step. The decrease of adsorption of AC1 to Chl a and Chl b due to the oxidation of acrylic acid proves the importance of π-π interaction.


Rapid Ca2+-dependent decrease of protein ubiquitination at synapses.
Protein ubiquitination has been implicated in the regulation of axonal growth and synaptic plasticity as well as in the pathogenesis of neurodegenerative diseases. Here we show that depolarization-dependent Ca2+ influx into synaptosomes produces a global, rapid (range of seconds), and reversible decrease of the ubiquitinated state of proteins, which correlates with the Ca2+-dependent dephosphorylation of several synaptic proteins. A similar general decrease in protein ubiquitination was observed in nonneuronal cells on Ca2+ entry induced by ionomycin. Both in synaptosomes and in nonneuronal cells, this decrease was blocked by FK506 (a calcineurin antagonist). Proteins whose ubiquitinated state was decreased include epsin 1, a substrate for the deubiquitinating enzyme fat facets/FAM, which we show here to be concentrated at synapses. These results reveal a fast regulated turnover of protein ubiquitination. In nerve terminals, protein ubiquitination may play a role both in the regulation of synaptic function, including vesicle traffic, and in the coordination of protein turnover with synaptic use.


Intervacuole exchange in the yeast zygote: a new pathway in organelle communication.
A new pathway of vesicle traffic between organelles has been identified. The vacuoles (lysosomes) of Saccharomyces cerevisiae zygotes rapidly exchange their contents at a specific point in the cell cycle. With the use of fluorescence microscopy, "tracks" were observed that connect the original parental vacuoles to the newly forming bud vacuoles. These observations suggest that vacuole-derived vesicles rapidly move along the tracks in both directions, equilibrating vacuole contents. This rapid vesicle movement may be responsible for vacuole formation in newly developing cells.


Bronchoconstrictor effects of leukotriene C in humans.
Maximum expiratory flow rate at 30 percent of vital capacity above residual volume served as an index of airway obstruction in comparing the effects of leukotriene C and histamine administered by aerosol to five normal persons. Leukotriene C was 600 to 9500 times more potent than histamine on a molar basis in producing an equivalent decrement in the residual volume. The leukotriene C response was slow in onset and prolonged, reminiscent of the effects of aerosol allergen challenge in asthmatic allergic subjects.


String tension and stability of magic tip-suspended nanowires.
Multishell helical gold nanowires were recently imaged by electron microscopy. We show theoretically that the contact with the gold tips at either end of the wire plays a crucial role and that local minima in the string tension rather than the total wire free energy determine the nanowire stability. Density functional electronic structure calculations of the simplest and thinnest coaxial gold and silver wires of variable radius and chirality were carried out. We found a string tension minimum for a single-tube gold nanowire that is chiral and consists of seven strands, in striking agreement with observation. In contrast, no such minimum was found for silver, where the s-d competition leading to surface reconstruction is lacking.


Protein folding cooperativity: basic insights from minimalist models.
Basic concepts about two-state, cooperative protein folding and its relation to first-order phase transitions are reviewed. Minimalist models capable of reproducing the required free energy barrier between folded and unfolded macroscopic states are described. A significantly more restrictive "calorimetric" criterion is also discussed, which is based on direct comparison between model and experimental heat capacities with additional assumptions about conformational enthalpy variation in the unfolded state.


Site-directed mutagenesis of the substrate-binding cleft of human estrogen sulfotransferase.
The sulfonation of estrogens by human estrogen sulfotransferase (humSULT1E1) plays a vital role in controlling the active levels of these hormones in the body. To understand more fully the structural and functional characteristics of humSULT1E1, we have carried out site-directed mutagenesis of critical amino acids found in the substrate-binding cleft. Three single amino acid mutations of humSULT1E1 (V145E, H107A, and K85A) were created in this study. Kinetic studies were used to provide information about the importance of these residues in substrate specificity and catalysis, using a variety of substrates. Lysine at position 85 has been proposed to be within hydrogen bonding distance to the 3alpha-phenol group of beta-estradiol, thereby stabilising the substrate in the active site. However, substitution to a neutral alanine at this position improved substrate specificity of humSULT1E1 for beta-estradiol, estrone, and dehydroepiandrosterone (DHEA). The exchange of valine 145 for negatively charged glutamic acid markedly improved the ability of humSULT1E1 to sulfonate dopamine, but caused a reduction in specificity constants toward steroids tested, in particular DHEA. The presence of a histidine residue at position 107 was shown to be essential for the production of a functional protein, as substitution of this amino acid to alanine resulted in complete loss of activity of humSULT1E1 towards all substrates tested.


Allometric escape and acoustic signal features facilitate high-frequency communication in an endemic Chinese primate.
The principle of acoustic allometry-the larger the animal, the lower its calls' fundamental frequency-is generally observed across terrestrial mammals. Moreover, according to the Acoustic Adaptation Hypothesis, open habitats favor the propagation of high-frequency calls compared to habitats with complex vegetational structures. We carried out playback experiments in which the calls of the Guizhou snub-nosed monkey (Rhinopithecus brelichi) were used as stimuli in sound attenuation and degradation experiments to test the hypothesis that propagation of Guizhou snub-nosed monkey calls is favored above vs through the forest floor vegetation. We found that low-pitched Guizhou snub-nosed monkey vocalizations suffered less attenuation than its high-pitched calls. Guizhou snub-nosed monkeys were observed emitting high-pitched calls from 1.5 to 5.0 m above the ground. The use of high-pitched calls from these heights coupled with the concomitant behavior of moving about above the understory may provide a signal for receivers which maximizes potential transmission and efficacy. Our results support the Acoustic Adaptation Hypothesis and suggest that by uncoupling its vocal output from its size, this monkey can produce a high-pitched call with a broad spectral bandwidth, thereby increasing both its saliency and the frequency range over which the animal may more effectively communicate in its natural habitat.


Medical Correlates of Chronic Multisymptom Illness in Gulf War Veterans.
Chronic multisymptom illness (CMI) is more prevalent among deployed than nondeployed veterans from the first Gulf War. Objective physiologic markers of CMI are lacking. The purpose of this study is to determine whether measurable abnormalities in the autonomic nervous system or hypothalamic-pituitary adrenal axis would distinguish CMI cases (CMI+) from controls (CMI-) among deployed veterans of the 1990-1991 Gulf War. This is a cross-sectional case-control cohort study that examined deployed veterans who participated in the Phase III study: National Health Survey of Gulf War Veterans and Their Families. Autonomic nervous system and hypothalamic-pituitary adrenal axis function-related measures included: 24-hour heart-rate variability, urinary catecholamines and cortisol, hypertension, insulin sensitivity, dyslipidemia, body fat, bone mineral density, and ultrasensitive C-reactive protein. Veterans of the first Gulf War with CMI (n = 73) and without the condition (n = 111) were studied. Sociodemographic characteristics were similar. Veterans with CMI reported poorer mental and physical functioning, greater use of prescription medications, and more nonroutine clinic visits. These veterans were also more likely to have fibromyalgia syndrome, irritable bowel syndrome, metabolic syndrome, and among males, a larger waist-to-hip ratio. Lower values for a nonlinear heart-rate-variability parameter-the short-term fractal scaling exponent (DFA1), reflecting an increased randomness of beat-to-beat changes in heart rate-were observed in veterans with CMI than those veterans without it (1.28±0.16vs 1.35±0.15; p=0.005). Hypothalamic-pituitary-adrenal axis function measures were similar between the two groups. In this cohort of deployed veterans from the first Gulf War, we identified abnormal heart-rate variability in veterans with CMI compared to veterans without the condition, which suggests abnormal functioning of the autonomic nervous system and possible long-term cardiovascular effects.


In situ monitoring of the nascent Pseudomonas fluorescens biofilm response to variations in the dissolved organic carbon level in low-nutrient water by attenuated total reflectance-Fourier transform infrared spectroscopy.
Drinking water quality management requires early warning tools which enable water supply companies to detect quickly and to forecast degradation of the microbial quality of drinking water during its transport throughout distribution systems. This study evaluated the feasibility of assessing, in real time, drinking water biostability by monitoring in situ the evolution of the attenuated total reflectance-Fourier transform infrared (ATR-FTIR) fingerprint of a nascent reference biofilm exposed to water being tested. For this purpose, the responses of nascent Pseudomonas fluorescens biofilms to variations in the dissolved organic carbon (DOC) level in tap water were monitored in situ and in real time by ATR-FTIR spectroscopy. Nascent P. fluorescens biofilms consisting of a monolayer of bacteria were formed on the germanium crystal of an ATR flowthrough cell by pumping bacterial suspensions in Luria-Bertani (LB) medium through the cell. Then they were exposed to a continuous flow of dechlorinated sterile tap water supplemented with appropriate amounts of sterile LB medium to obtain DOC concentrations ranging from 1.5 to 11.8 mg/liter. The time evolution of infrared bands related to proteins, polysaccharides, and nucleic acids clearly showed that changes in the DOC concentration resulted in changes in the nascent biofilm ATR-FTIR fingerprint within 2 h after exposure of the biofilm to the water being tested. The initial bacterial attachment, biofilm detachment, and regrowth kinetics determined from changes in the areas of bands associated with proteins and polysaccharides were directly dependent on the DOC level. Furthermore, they were consistent with bacterial adhesion or growth kinetic models and extracellular polymeric substance overproduction or starvation-dependent detachment mechanisms.


Cerebral Dynamics during the Observation of Point-Light Displays Depicting Postural Adjustments.
Objective: As highly social creatures, human beings rely part of their skills of identifying, interpreting, and predicting the actions of others on the ability of perceiving biological motion. In the present study, we aim to investigate the electroencephalographic (EEG) cerebral dynamics involved in the coding of postural control and examine whether upright stance would be codified through the activation of the temporal-parietal cortical network classically enrolled in the coding of biological motion. Design: We registered the EEG activity of 12 volunteers while they passively watched point light displays (PLD) depicting quiet stable (QB) and an unstable (UB) postural situations and their respective scrambled controls (QS and US). In a pretest, 13 volunteers evaluated the level of stability of our two biological stimuli through a stability scale. Results: Contrasting QB vs. QS revealed a typical ERP difference in the right temporal-parietal region at an early 200-300 ms time window. Furthermore, when contrasting the two biological postural conditions, UB vs. QB, we found a higher positivity in the 400-600 ms time window for the UB condition in central-parietal electrodes, lateralized to the right hemisphere. Conclusions: These results suggest that PLDs depicting postural adjustments are coded in the brain as biological motion, and that their viewing recruit similar networks with those engaged in postural stability control. Additionally, higher order cognitive processes appear to be engaged in the identification of the postural instability level. Disentangling the EEG dynamics during the observation of postural adjustments could be very useful for further understanding the neural mechanisms underlying postural control.


Conjugated equine estrogens and incidence of probable dementia and mild cognitive impairment in postmenopausal women: Women's Health Initiative Memory Study.
The Women's Health Initiative Memory Study (WHIMS) previously found increased risk for dementia and no effect on mild cognitive impairment (MCI) in women treated with conjugated equine estrogens (CEE) plus medroxyprogesterone acetate (MPA). To determine the effects of CEE alone and CEE plus MPA on incidence of probable dementia and MCI in older women. Randomized, double-blind, placebo-controlled clinical trials of CEE (estrogen-alone trial) or CEE plus MPA (estrogen plus progestin trial) in community-dwelling women aged 65 to 79 years, conducted from June 1995 to July 8, 2002 (estrogen plus progestin; n = 4532), or to February 29, 2004 (estrogen-alone; n = 2947), in 39 of the 40 WHI clinical centers. In the estrogen-alone trial, 1 daily tablet containing either 0.625 mg/d of CEE vs matching placebo; in the estrogen plus progestin trial, 1 daily tablet containing CEE (0.625 mg/d) plus MPA (2.5 mg/d) vs matching placebos. Probable dementia and MCI. In the estrogen-alone trial, 47 participants were diagnosed with probable dementia, of whom 28 were assigned to CEE and 19 to placebo (hazard ratio [HR], 1.49; 95% confidence interval [CI], 0.83-2.66). Incidence rates for probable dementia in the estrogen-alone trial were statistically similar to those in the estrogen plus progestin trial (45 vs 22 per 10 000 person-years for CEE plus MPA vs placebo, respectively; P =.11). When data were pooled per the original WHIMS protocol, the overall HR for probable dementia was 1.76 (95% CI, 1.19-2.60; P =.005). After excluding participants with baseline Modified Mini-Mental State Examination scores at or below the screening cut point, the HR was 1.77 (95% CI, 0.74-4.23; P =.20) in the estrogen-alone trial and 2.19 (95% CI, 1.25-3.84; P =.006) in the pooled trials. In the estrogen-alone trial, 76 participants were diagnosed with MCI in the CEE group vs 58 in the placebo group (HR, 1.34; 95% CI, 0.95-1.89). In the combined trial data, the HR was similar (1.25; 95% CI, 0.97-1.60). In the estrogen-alone trial, 93 participants receiving CEE were diagnosed with either probable dementia or MCI vs 69 receiving placebo (HR, 1.38; 95% CI, 1.01-1.89; P =.04). Estrogen therapy alone did not reduce dementia or MCI incidence and increased the risk for both end points combined. Pooling data for estrogen alone and estrogen plus progestin resulted in increased risks for both end points. Use of hormone therapy to prevent dementia or cognitive decline in women 65 years of age or older is not recommended.


Contribution of serum biomarkers to the diagnosis of invasive candidiasis.
Invasive candidiasis is the most important opportunistic fungal infection, causing high morbidity and mortality. Traditional methods of diagnosis, which include blood culture and biopsy, usually lack both sensitivity and specificity, or become positive late in the course of the infection. Therefore, new nonculture-based methods are being developed. In this review, we will discuss the most recent studies concerning the use of serum biomarkers in the diagnosis of invasive Candida infections.


Effects of food deprivation and refeeding on total protein and actomyosin degradation.
Total protein and actomyosin degradation rates were determined in perfused rat hemicorpus preparations. By simultaneously measuring the release of two nonmetabolizable amino acids phenylalanine and N tau-methylhistidine from the hemicorpus, the respective rates of total protein and actomyosin degradation could be calculated. When rats were deprived of food for 48 h, the rate of total protein degradation increased to 148% of the fed controls. If rats were food deprived and then refed for 24 h, the degradation rate decreased to only 79% of the rate of fed controls. Measurement of N tau-methylhistidine release indicated that food deprivation led to a dramatic increase in the rate of actomyosin degradation (427% of fed), whereas refeeding decreased the actomyosin degradation rate to that of the fed controls. Calculations of the fractional degradation rates show that actomyosin breaks down at a much slower rate than the nonactomyosin proteins (1.5 vs. 20.8%/day in preparations from fed rats, and 6.2 vs. 28.2%/day in preparations from food-deprived rats). Therefore, the contribution of actomyosin breakdown to total muscle protein breakdown is small in the fed state (11%) and increased threefold after food deprivation. The addition of insulin to the perfusion medium decreased the rate of total protein degradation by 18% in preparations from food-deprived rats with no significant effect on actomyosin degradation. Thus, in vitro, insulin's major effect may be to decrease the degradation of more rapidly turning over, nonactomyosin proteins. Protein degradation, as well as protein synthesis, contributes to the adaptation of muscle to starvation and refeeding.(ABSTRACT TRUNCATED AT 250 WORDS)


Glyoxal oxidases: their nature and properties.
H2O2 has been found to be required for the activity of the main microbial enzymes responsible for lignin oxidative cleavage, peroxidases. Along with other small radicals, it is implicated in the early attack of plant biomass by fungi. Among the few extracellular H2O2-generating enzymes known are the glyoxal oxidases (GLOX). GLOX is a copper-containing enzyme, sharing high similarity at the level of active site structure and chemistry with galactose oxidase. Genes encoding GLOX enzymes are widely distributed among wood-degrading fungi especially white-rot degraders, plant pathogenic and symbiotic fungi. GLOX has also been identified in plants. Although widely distributed, only few examples of characterized GLOX exist. The first characterized fungal GLOX was isolated from Phanerochaete chrysosporium. The GLOX from Utilago maydis has a role in filamentous growth and pathogenicity. More recently, two other glyoxal oxidases from the fungus Pycnoporus cinnabarinus were also characterized. In plants, GLOX from Vitis pseudoreticulata was found to be implicated in grapevine defence mechanisms. Fungal GLOX were found to be activated by peroxidases in vitro suggesting a synergistic and regulatory relationship between these enzymes. The substrates oxidized by GLOX are mainly aldehydes generated during lignin and carbohydrates degradation. The reactions catalysed by this enzyme such as the oxidation of toxic molecules and the production of valuable compounds (organic acids) makes GLOX a promising target for biotechnological applications. This aspect on GLOX remains new and needs to be investigated.


The mitochondrial adenosine 5'-triphosphatase in slow and fast heart rate hearts.
A survey of 12 species has revealed that reversible ischemia-induced protonic inhibition of the cardiac muscle mitochondrial adenosine 5'-triphosphatase (ATPase) described by this author earlier (Rouslin, W. J. Biol. Chem. 258: 9657-9661, 1983) occurs only in animals with heart rates lower than approximately 200 beats/min. It was thus fully demonstrable in rabbit, dog, sheep, human, pig, and beef heart mitochondria. In contrast, the in situ ATPase inhibition was completely absent in six smaller species capable of heart rates of approximately 300 or more beats/min. These were chicken, pigeon, guinea pig, rat, hamster, and mouse. Analyses of the cardiac muscle mitochondria of 9 of the 12 species studied showed them to contain normal levels of mitochondrial ATPase inhibitor; the three smallest species, rat, hamster, and mouse contained only very low levels of inhibitor. Thus, although chicken, pigeon, and guinea pig heart mitochondria contained normal levels of ATPase inhibitor, they (like the rat, hamster, and mouse) showed no in situ ischemia-induced ATPase inhibition. This and other observations suggest that the lack of in situ ATPase inhibition in hearts capable of 300 or more beats/min may be due to the presence of either an in situ nonfunctional ATPase inhibitor protein or to an in situ uninhibitable form of the mitochondrial ATPase in the faster-paced hearts. Alternatively, the mitochondria of the fast-paced hearts may be insulated somehow against the cytosolic acidosis which develops during ischemia and which triggers the ATPase inhibition in the slow heart-rate hearts. In the faster paced hearts, ATP hydrolysis does not appear to be regulated by inhibitor binding to the ATPase under nonenergizing conditions.


A degradable soybean-based biomaterial used effectively as a bone filler in vivo in a rabbit.
The 'gold standard' for bone filling is currently the bone autograft, but its use is limited by material availability and by the possible risks of infection or other donor site morbidity. Materials proposed so far as bone fillers do not show all the characteristics which are desirable. These are (a) osteoconductivity, (b) controlled biodegradation and (c) ease of adaptation to the implantation site. Recently, a new class of biodegradable material based on soybeans has been presented which shows good mechanical properties and an intrinsic bioactivity on inflammatory and tissue cells in vitro. The authors investigated the morphology in vivo of bone response in repairing a surgical lesion in the presence of granules of a novel soybean-based biomaterial (SB), comparing it with a sham-operated contralateral lesion of critical size (non-healing model); 26 operations were performed in New Zealand White rabbits, with back scattered electron microscopy as the analysis technique of choice. Implantation of SB granules over 8 weeks produced bone repair with features distinct from those obtained by healing in a non-treated defect. New and progressively maturing trabeculae appeared in the animal group where SB granules were implanted, while sham operation produced only a rim of pseudo-cortical bone still featuring a large defect. The trabeculae forming in the presence of SB granules had features typical of reticular bone. These findings suggest that the bone regeneration potential of SB granules and their intrinsic bioactivity, combined with their relatively easy and cost-effective preparation procedures, make them suitable candidates as a bone filler in clinical applications.


Comparison of Two Point-of-Care CYP2C19 Genotyping Assays for Genotype-Guided Antiplatelet Therapy.
CYP2C19 polymorphisms contribute about 12% of the variability in the antiplatelet effect of clopidogrel, which is commonly prescribed for patients undergoing percutaneous coronary intervention. For these patients, rapid turnaround time of CYP2C19 genotyping may be critical. We validated and compared the performance of two point-of-care CYP2C19 genotype tests, Nanosphere Verigene CYP2C19 Nucleic Acid Test and Spartan RX CYP2C19 System. Our CLIA certified Molecular Diagnostic Laboratory performed 99 Verigene tests and 108 Spartan RX CYP2C19 assays. We compared performance and genotype results between the two platforms, across runs, and among technologists. Based on our validation results, we started offering CYP2C19 genotyping using the Spartan RX CYP2C19 assay for post-percutaneous coronary intervention patients. Laboratory validation genotype results were consistent between both assays when the assays produced results (100% accuracy); however, the Verigene CYP2C19 had a 33% no call rate. In contrast, Spartan consistently showed accurate results. Using a newly established clinical workflow, we assayed 342 post-percutaneous coronary intervention patients with the Spartan test. Within one hour of submitting patient samples, ordering physicians were notified of any clinically significant results and provided clinical decision support. Every approach has its limitations, but our practice of using the Spartan RX CYP2C19 test in our acute cardiac workflow provides accurate and rapid results to guide clinical decision-making at the point-of-care. Prospective follow-up is ongoing to evaluate outcomes and effectiveness of CYP2C19 testing.


Water encapsulation in a polyoxapolyaza macrobicyclic compound.
A new heteroditopic macrobicyclic compound (t(2)pN(5)O(3)) containing two separate polyoxa and polyaza compartments was synthesized in good yield through a [1 + 1] "tripod-tripod coupling" strategy. The X-ray crystal structure of H(3)t(2)pN(5)O(3)(3+) revealed the presence of one encapsulated water molecule accepting two hydrogen bonds from two protonated secondary amines and donating a hydrogen bond to one amino group. The acid-base behavior of the compound was studied by potentiometry at 298.2 K in aqueous solution and at ionic strength 0.10 M in KCl. The results revealed unusual protonation behavior, namely a surprisingly low fourth protonation constant contrary to what was expected for the compound. (1)H NMR and DOSY experiments, as well as molecular modeling studies, showed that the water encapsulation and the conformation observed in the solid state are retained in solution. The strong binding of the encapsulated water molecule, reinforced by the cooperative occurrence of a trifurcated hydrogen bond at the polyether compartment of the macrobicycle, account for the very low log K(4)(H) value obtained.


The Saccharomyces cerevisiae enolase-related regions encode proteins that are active enolases.
In addition to two genes (ENO1 and ENO2) known to code for enolase (EC18.104.22.168), the Saccharomyces cerevisiae genome contains three enolase-related regions (ERR1, ERR2 and ERR3) which could potentially encode proteins with enolase function. Here, we show that products of these genes (Err2p and Err3p) have secondary and quaternary structures similar to those of yeast enolase (Eno1p). In addition, Err2p and Err3p can convert 2-phosphoglycerate to phosphoenolpyruvate, with kinetic parameters similar to those of Eno1p, suggesting that these proteins could function as enolases in vivo. To address this possibility, we overexpressed the ERR2 and ERR3 genes individually in a double-null yeast strain lacking ENO1 and ENO2, and showed that either ERR2 or ERR3 could complement the growth defect in this strain when cells are grown in medium with glucose as the carbon source. Taken together, these data suggest that the ERR genes in Saccharomyces cerevisiae encode a protein that could function in glycolysis as enolase. The presence of these enolase-related regions in Saccharomyces cerevisiae and their absence in other related yeasts suggests that these genes may play some unique role in Saccharomyces cerevisiae. Further experiments will be required to determine whether these functions are related to glycolysis or other cellular processes.


Fracture prediction for the proximal femur using finite element models: Part II--Nonlinear analysis.
In Part I we reported the results of linear finite element models of the proximal femur generated using geometric and constitutive data collected with quantitative computed tomography. These models demonstrated excellent agreement with in vitro studies when used to predict ultimate failure loads. In Part II, we report our extension of those finite element models to include nonlinear behavior of the trabecular and cortical bone. A highly nonlinear material law, originally designed for representing concrete, was used for trabecular bone, while a bilinear material law was used for cortical bone. We found excellent agreement between the model predictions and in vitro fracture data for both the onset of bone yielding and bone fracture. For bone yielding, the model predictions were within 2 percent for a load which simulated one-legged stance and 1 percent for a load which simulated a fall. For bone fracture, the model predictions were within 1 percent and 17 percent, respectively. The models also demonstrated different fracture mechanisms for the two different loading configurations. For one-legged stance, failure within the primary compressive trabeculae at the subcapital region occurred first, leading to load transfer and, ultimately, failure of the surrounding cortical shell. However, for a fall, failure of the cortical and trabecular bone occurred simultaneously within the intertrochanteric region. These results support our previous findings that the strength of the subcapital region is primarily due to trabecular bone whereas the strength of the intertrochanteric region is primarily due to cortical bone.


Titanium elastic nail - Complications in the treatment of paediatric diaphyseal fracture of femur [corrected].
Femoral shaft fracture is the most common major paediatric orthopaedic. For generations traction and casting was the standard treatment for all femoral shaft fractures in children. Over the past two decades the advantages of fixation and rapid mobilisation have been increasingly recognised. A prospective study was conducted in five private hospitals in the district of Bankura, West Bengal over a period of two years (April 2010 to March 2012) on 70 patients with closed shaft femur fractures between 6- 14 years age of either sex. The aim was to find out the short term complications of titanium elastic nailing in diaphyseal fracture of femur in children and compare the findings of this study with pre- existing studies in this field. In our study the most common complication was pain at nail entry site (60%). 5.71% had local inflammatory reaction due to nails. Superficial infection occurred in 2.85%. At the end of 1 year, 2.85% had limb length discripancies. Proximal migration occurred in 2.85%. 2.85% encountered acute reactive synovitis, 5.71% developed angulation of fracture site and 2.85% developed per operative breakage of nail. The treatment of paediatric shaft femur fracture has been drastically changed over the last two decades to internal fixation by elastically stable intra- medullary nail (ESIN). In our study, we encountered only a few complications most of them being minor. Most of the complications were surgical technique related and were seen at the initial phase of the learning curve.


Plasmacytoid dendritic cell biology and its role in immune-mediated diseases.
Plasmacytoid dendritic cells (pDCs) are a unique subset of dendritic cells specialised in secreting high levels of type I interferons. pDCs play a crucial role in antiviral immunity and have been implicated in the initiation and development of many autoimmune and inflammatory diseases. This review summarises the latest advances in recent years in several aspects of pDC biology, with special focus on pDC heterogeneity, pDC development via the lymphoid pathway, and newly identified proteins/pathways involved in pDC trafficking, nucleic acid sensing and interferon production. Finally, we also highlight the current understanding of pDC involvement in autoimmunity and alloreactivity, and opportunities for pDC-targeting therapies in these diseases. These new insights have contributed to answers to several fundamental questions remaining in pDC biology and may pave the way to successful pDC-targeting therapy in the future.


Evaluation of quality of sleep and effecting factors in hospitalized neurosurgical patients.
To evaluate the quality of sleep and effecting factors in hospitalized patients in a neurosurgery clinic. This research was conducted as a descriptive study in a neurosurgery clinic of a University Hospital, in Turkey, between November 2005 and June 2006. The study sample consisted of 102 patients who met the inclusion criteria and agreed to participate in the study. Data were collected using the "Questionnaire Form," which included socio-demographic, illness, sleep, and hospital characteristics, and the "Pittsburgh Sleep Quality Index" (PSQI). The data obtained were assessed by SPSS 10.00 Program, using number, percentage, arithmetic mean, t-test, variance, and correlation analysis. The mean age of the patients was 43.22+/-16.72 years, 54.9% were females, and 72.5% were married. Hospitalization duration of the patients was 14.31+/-8.97 days on average, 84.3% had previous hospital experience, and 72.5% had knowledge about their illnesses. Of them, 51% expressed a sleep problem. Pain (41.2%) and noise (31.4%) was found to affect the sleep quality of hospitalized patients. Of the patients hospitalized in the neurosurgery ward, 64.7% were found to have poor quality of sleep prior to admission, while poor quality of sleep was determined for 49% before being discharged from the hospital. However, a better sleep quality was noted prior to discharge, compared to that at their admission to the hospital. Varied degrees of quality of sleep have been reported by the patients and influences of personal and environmental factors on sleep quality have been noted. Strategies to enhance physical and mental comfort, and to minimize disruptions should positively enhance sleep quality.


Mechanosensitive channels and bacterial cell wall integrity: does life end with a bang or a whimper?
Mechanogated channels are fundamental components of bacterial cells that enable retention of physical integrity during extreme increases in cell turgor. Optical tweezers combined with microfluidics have been used to study the fate of individual Escherichia coli cells lacking such channels when subjected to a bursting stress caused by increased turgor. Fluorescence-activated cell sorting and electron microscopy complement these studies. These analyses show that lysis occurs with a high probability, but the precise path differs between individual cells. By monitoring the loss of cytoplasmic green fluorescent protein, we have determined that some cells release this protein but remain phase dark (granular) consistent with the retention of the majority of large proteins. By contrast, most cells suffer cataclysmic wall failure leading to loss of granularity but with the retention of DNA and overall cell shape (protein-depleted ghosts). The time span of these events induced by hypo-osmotic shock varies but is of the order of milliseconds. The data are interpreted in terms of the timing of mechanosensitive channel gating relative to osmotically induced water influx.


Immunoprofiling using NAPPA protein microarrays.
Protein microarrays provide an efficient method to immunoprofile patients in an effort to rapidly identify disease immunosignatures. The validity of using autoantibodies in diagnosis has been demonstrated in type 1 diabetes, rheumatoid arthritis, and systemic lupus, and is now being strongly considered in cancer. Several types of protein microarrays exist including antibody and antigen arrays. In this chapter, we describe the immunoprofiling application for one type of antigen array called NAPPA (nucleic acids programmable protein array). We provide a guideline for setting up the screening study and designing protein arrays to maximize the likelihood of obtaining quality data.


Identification of a novel UDP-Gal:GalNAc beta 1-4GlcNAc-R beta 1-3-galactosyltransferase in the connective tissue of the snail Lymnaea stagnalis.
Connective tissue of the freshwater pulmonate Lymnaea stagnalis was shown to contain galactosyltransferase activity capable of transferring Gal from UDP-Gal in beta 1-3 linkage to terminal GalNAc of GalNAc beta 1-4GlcNAc-R [R = beta 1-2Man alpha 1-O(CH2)8COOMe, beta 1-OMe, or alpha,beta 1-OH]. Using GalNAc beta 1-4GlcNAc beta 1-2Man alpha-1-O(CH2)8COOMe as substrate, the enzyme showed an absolute requirement for Mn2+ with an optimum Mn2+ concentration between 12.5 mM and 25 mM. The divalent cations Mg2+, Ca2+, Ba2+ and Cd2+ at 12.5 mM could not substitute for Mn2+. The galactosyltransferase activity was independent of the concentration of Triton X-100, and no activation effect was found. The enzyme was active with GalNAc beta 1-4GlcNAc beta 1-2Man alpha 1-O(CH2)8COOMe (Vmax 140 nmol.h-1.mg protein-1; Km 1.02 mM), GalNAc beta 1-4GlcNAc (Vmax 105 nmol.h-1.mg protein-1; Km 0.99 mM), and GalNAc beta 1-4GlcNAc beta 1-OMe (Vmax 108 nmol.h-1.mg protein-1; Km 1.33 mM). The products formed from GalNAc beta 1-4GlcNAc beta 1-2Man alpha 1-O(CH2)8COOMe and GalNAc beta 1-4GlcNAc beta 1-OMe were purified by high performance liquid chromatography, and identified by 500-MHz 1H-NMR spectroscopy to be Gal beta 1-3GalNAc beta 1-4GlcNAc 1-OMe, respectively. The enzyme was inactive towards GlcNAc, GalNac beta 1-3 GalNAc alpha 1-OC6H5, GalNAc alpha 1--ovine-submaxillary-mucin, lactose and N-acetyllactosamine. This novel UDP-Gal:GalNAc beta 1-4GlcNAc-R beta 1-3-galactosyltransferase is believed to be involved in the biosynthesis of the hemocyanin glycans of L. stagnalis.


The effect of dietary supplementation with Aurantiochytrium limacinum on lactating dairy cows in terms of animal health, productivity and milk composition.
The aim of this study was to investigate the effect of dietary supplementation with the docosahexaenoic acid (DHA)-rich microalgae, Aurantiochytrium limacinum (AURA), on a variety of health and productivity parameters in lactating cows. Twenty-four cows were blocked by parity and number of days in milk and then randomly assigned to a control (CON; n = 12) group with no algal supplementation, or a treatment group (AURA; n = 12) provided with 100 g AURA cow-1 day-1 or 16 g DHA cow-1 day-1 . A variety of health and productivity measurements were taken, and results indicated that supplementation had no negative effects on animal health in terms of somatic cell count, haematological and biochemical blood parameters, while body condition was marginally improved by algal supplementation. No differences were found for the various production parameters measured; however, a tendency towards increased milk production was observed for the AURA group during the final stage of the study (+4.5 kg cow-1 day-1 , day 78-84). The fatty acid profile of milk was improved by supplementation, with significantly lower saturated fatty acids, significantly higher omega-3 fatty acids and an improved omega-3/omega-6 ratio observed when compared to the control group. The amount of DHA in the milk of cows provided 105 g AURA head-1 day-1 was 4.7 mg/100 g milk with a peak transfer efficiency from feed to milk at day 49 of 8.3%. These results indicate that supplementation with 105 g AURA head-1 day-1 resulted in the successful enrichment of milk with DHA without negatively impacting the health or productivity of the animals.


The design and construction of a realistic thorax phantom for in vivo measurements of low-energy photon emitters.
Calibration of detector systems for the in vivo measurement of transuranic radionuclides is notoriously difficult due to the low energies and intensities of the photon emissions and their severe attenuation within the tissues of the body. A realistic chest phantom has been designed and constructed to provide calibration data to relate detector responses to lung contents of radionuclides; the phantom incorporates a human thoracic cage and tissue substitutes that have been carefully selected to reproduce the scattering and absorption coefficients of muscle and lung tissue. The measured attenuation coefficients of the substitutes are compared with values calculated for the tissues of interest. Preliminary calibration data obtained from this phantom are presented and the uses and limitations of this calibration procedure are discussed.


Locomotor muscle profile of a deep (Kogia breviceps) versus shallow (Tursiops truncatus) diving cetacean.
When a marine mammal dives, breathing and locomotion are mechanically uncoupled, and its locomotor muscle must power swimming when oxygen is limited. The morphology of that muscle provides insight into both its oxygen storage capacity and its rate of oxygen consumption. This study investigated the m. longissimus dorsi, an epaxial swimming muscle, in the long duration, deep-diving pygmy sperm whale (Kogia breviceps) and the short duration, shallow-diving Atlantic bottlenose dolphin (Tursiops truncatus). Muscle myoglobin content, fiber type profile (based upon myosin ATPase and succinate dehydrogenase assays), and fiber size were measured for five adult specimens of each species. In addition, a photometric analysis of sections stained for succinate dehydrogenase was used to create an index of mitochondrial density. The m. longissimus dorsi of K. breviceps displayed significantly a) higher myoglobin content, b) larger proportion of Type I (slow oxidative) fibers by area, c) larger mean fiber diameters, and d) lower indices of mitochondrial density than that of T. truncatus. Thus, this primary swimming muscle of K. breviceps has greater oxygen storage capacity, reduced ATP demand, and likely a reduced rate of oxygen consumption relative to that of T. truncatus. The locomotor muscle of K. breviceps appears able to ration its high onboard oxygen stores, a feature that may allow this species to conduct relatively long duration, deep dives aerobically.


PTEN-inducible kinase 1 (PINK1)/Park6 is indispensable for normal heart function.
Oxidative stress is caused by an imbalance between reactive oxygen species (ROS) production and the ability of an organism to eliminate these toxic intermediates. Mutations in PTEN-inducible kinase 1 (PINK1) link mitochondrial dysfunction, increased sensitivity to ROS, and apoptosis in Parkinson's disease. Whereas PINK1 has been linked to the regulation of oxidative stress, the exact mechanism by which this occurs has remained elusive. Oxidative stress with associated mitochondrial dysfunction leads to cardiac dysfunction and heart failure (HF). We hypothesized that loss of PINK1 in the heart would have deleterious consequences on mitochondrial function. Here, we observed that PINK1 protein levels are markedly reduced in end-stage human HF. We also report that PINK1 localizes exclusively to the mitochondria. PINK1(-/-) mice develop left ventricular dysfunction and evidence of pathological cardiac hypertrophy as early as 2 mo of age. Of note, PINK1(-/-) mice have greater levels of oxidative stress and impaired mitochondrial function. There were also higher degrees of fibrosis, cardiomyocyte apoptosis, and a reciprocal reduction in capillary density associated with this baseline cardiac phenotype. Collectively, our in vivo data demonstrate that PINK1 activity is crucial for postnatal myocardial development, through its role in maintaining mitochondrial function, and redox homeostasis in cardiomyocytes. In conclusion, PINK1 possesses a distinct, nonredundant function in the surveillance and maintenance of cardiac tissue homeostasis.


Isolated Limb Perfusion on Nonmelanoma Skin Cancer for Limb Salvage: A Series of Four Cases.
Nonmelanoma skin cancer (NMSC) is more prevalent than all the other cancers combined together. The most common regions affected by skin cancer are the head and neck, but there is a large proportion of the cases located on the limbs. They could be bulky, very extensive and/or located in specialized regions like the hands or near to a joint. Most of those cases should be amputated, with several compromises of the function and a negative impact on the patients' quality of life. Isolated limb perfusion is a proven alternative to limb salvage on soft tissue sarcomas, but there are just a few reports about its application on non-melanoma skin cancer. The aim of the article is to describe the outcomes and prove the benefits and effectiveness to avoid limb amputation when using isolated limb perfusion on locally advanced non-melanoma skin cancer. We present clinical, retrospective study as a case series report. The study includes four patients with locally advanced non-melanoma skin cancer in the limb - three cases with squamous cell carcinoma (SCC), and one patient with Merkel cell carcinoma (MCC) who underwent to tumor necrosis factor-alpha (TNFα) and melphalan based isolated perfusion of the limb (TM-ILP). Toxicity, clinical response, and limb salvage were described. Patients were treated in the oncological surgical department in a referral hospital in Mexico. The limb salvage rate was achieved in 75%. All patients had a favorable response to TM-ILP. Complete response of the tumor was noted on two (50%) of the patients (one with histopathological confirmation), the other 50% had a partial response to the TM-ILP. No serious toxicity related to TM-ILP was observed. One patient (25%) developed regional lymph involvement six months after perfusion. Lymphadenectomy was performed, and the patient subsequently has a six-year disease-free survival. In conclusion, TM-ILP could be an effective, reliable, and safe therapy on limb salvage in selected patients with non-melanoma skin cancer who are not adequate surgical candidates.


HIV serostatus disclosure among people living with HIV/AIDS in Mwanza, Tanzania.
Disclosing HIV serostatus is important for HIV prevention and maintenance of health for people living with HIV their spouses and the community, it plays a role in the social relation which is critical in reducing HIV transmission. The process may have positive and negative effects to the HIV infected people who disclose their status. The present study was undertaken to describe HIV serostatus disclosure among HIV infected people attending care and treatment clinic at Sekou-Toure hospital in Mwanza, Tanzania. A cross-sectional study was carried out on 270 HIV infected adults attending Care and Treatment Clinic (CTC) at Sekou-Toure hospital between September and October, 2010. A Swahili questionnaire was used to obtain demographic and HIV disclosure information. Hundred and ninety five (72.5%) of all recruited participants were females, 88.1% (238/270) were aged above 30 years and 44.1% (119/270) were married. The prevalence of serostatus disclosure was 93.3% (252/270) with participants aged above 30 years having significantly higher proportion of serostatus disclosure compared to those aged below 30 years (94.5% vs. 84.4%, p < 0.05). Among the participants who disclosed their status, 69.3% reported closeness to the disclosed person as the reason for disclosure while 25.8% (65/252) disclosed because they needed help. Two hundred (79.4%) reported to have received emotional support following disclosure while 25.8% and 29.7% received financial support and freedom to use their anti-retroviral drugs around the person they disclosed their status respectively. Thirty four participants reported to have been discriminated following disclosure and 12 participants reported to have been divorced. Rate of disclosure of HIV serostatus was noted to be high in this study. Delayed disclosure was also noted in small proportion of participants. Negative outcomes following disclosure of serostatus were reported by participants. Efforts need to be increased to promote disclosure of HIV serostatus in Tanzania through health education and awareness for both HIV infected individuals and the community.


Transactivation of cell death signals by glutamate transmission in dopaminergic neurons.
The intrinsic susceptibility of dopaminergic neurons underlies the pathophysiology of Parkinson's disease and is possibly related to developmental injury in schizophrenia. However, the molecular substrates for this susceptibility are not well understood. We review the evidence of selective susceptibility of dopaminergic neurons to excessive glutamate receptor stimulation and discuss the molecular pathways that differentiate between physiological and pathological signaling leading to this particular form of neuronal death. In vitro as well as in vivo, activation of GluRAMPA causes concentration-dependent, severe pruning of neurites and selective death of dopaminergic neurons. In primary cultures of mesencephalon, this form of injury is mediated through release of calcium from intracellular stores (CICR), leading to loss of calcium homeostasis, oxidative stress, and activation of the transcription factor NFkappaB and the cell death protein p53. Post-translational modification of p53 may be an important target for neuroprotection in Parkinson's disease and perhaps in prevention of other neuropsychiatric disorders.


Role of gamma-secretase in human umbilical-cord derived mesenchymal stem cell mediated suppression of NK cell cytotoxicity.
Mesenchymal stem cells (MSCs) are increasingly considered to be used as biological immunosuppressants in hematopoietic stem cell transplantation (HSCT). In the early reconstitution phase following HSCT, natural killer (NK) cells represent the major lymphocyte population in peripheral blood and display graft-vs-leukemia (GvL) effects. The functional interactions between NK cells and MSCs have the potential to influence the leukemia relapse rate after HSCT. Until date, MSC-NK cell interaction studies are largely focussed on bone marrow derived (BM)-MSCs. Umbilical cord derived (UC)-MSCs might be an alternative source of therapeutic MSCs. Thus, we studied the interaction of UC-MSCs with unstimulated allogeneic NK cells. UC-MSCs could potently suppress NK cell cytotoxicity in overnight cultures via soluble factors. The main soluble immunosuppressant was identified as prostaglandin (PG)-E2. Maximal PGE2 release involved IL-1β priming of MSCs after close contact between the NK cells and UC-MSCs. Interestingly, blocking gamma-secretase activation alleviated the immunosuppression by controlling PGE2 production. IL-1 receptor activation and subsequent downstream signalling events were found to require gamma-secretase activity. Although the role of PGE2 in NK cell-MSC has been reported, the requirement of cell-cell contact for PGE2 induced immunosuppression remained unexplained. Our findings shed light on this puzzling observation and identify new players in the NK cell-MSC crosstalk.


The lipid composition of Legionella dumoffii membrane modulates the interaction with Galleria mellonella apolipophorin III.
Apolipophorin III (apoLp-III), an insect homologue of human apolipoprotein E (apoE), is a widely used model protein in studies on protein-lipid interactions, and anti-Legionella activity of Galleria mellonella apoLp-III has been documented. Interestingly, exogenous choline-cultured Legionella dumoffii cells are considerably more susceptible to apoLp-III than non-supplemented bacteria. In order to explain these differences, we performed, for the first time, a detailed analysis of L. dumoffii lipids and a comparative lipidomic analysis of membranes of bacteria grown without and in the presence of exogenous choline. (31)P NMR analysis of L. dumoffii phospholipids (PLs) revealed a considerable increase in the phosphatidylcholine (PC) content in bacteria cultured on choline medium and a decrease in the phosphatidylethanolamine (PE) content in approximately the same range. The interactions of G. mellonella apoLp-III with lipid bilayer membranes prepared from PLs extracted from non- and choline-supplemented L. dumoffii cells were examined in detail by means of attenuated total reflection- and linear dichroism-Fourier transform infrared spectroscopy. Furthermore, the kinetics of apoLp-III binding to liposomes formed from L. dumoffii PLs was analysed by fluorescence correlation spectroscopy and fluorescence lifetime imaging microscopy using fluorescently labelled G. mellonella apoLp-III. Our results indicated enhanced binding of apoLp-III to and deeper penetration into lipid membranes formed from PLs extracted from the choline-supplemented bacteria, i.e. characterized by an increased PC/PE ratio. This could explain, at least in part, the higher susceptibility of choline-cultured L. dumoffii to G. mellonella apoLp-III.


Clinical characteristics, management, and outcome of incidental pulmonary embolism in cancer patients.
Incidental pulmonary embolisms (IPEs) are common in cancer patients. Examining the characteristics and outcomes of IPEs in cancer patients can help to ensure proper management, promoting better outcomes. To determine the clinical characteristics, management, and outcomes of IPEs for cancer patients, we conducted a 1:2 ratio case-control study and identified all consecutive patients with IPE who visited the emergency department at The University of Texas MD Anderson Cancer Center between 1 January 2006 and 1 January 2016. Each IPE case was matched with 2 controls using a propensity score obtained using logistic regression for IPE status with other factors affecting overall survival. A total of 904 confirmed cases were included in the analysis. IPE frequently occurred during the first year after cancer diagnosis (odds ratio [OR], 2.79; 95% confidence interval [95% CI], 2.37-3.29; P < .001). Patients receiving cytotoxic chemotherapy had a nearly threefold greater risk of developing IPE (OR, 2.87; 95% CI, 2.42-3.40; P < .001). In-hospital mortality was 1.9%. The 7- and 30-day mortality rates among the cases were 1.8% and 9.9%, respectively, which was significantly higher than in the control groups: 0.2% and 3.1%, respectively (P < .001). IPE was associated with reduced overall survival (hazard ratio [HR], 1.93; 95% CI, 1.74-2.14; P < .001). Concurrent incidental venous thromboembolism was identified in 189 of the patients (20.9%) and was also associated with reduced overall survival (HR, 1.65; 95% CI, 1.21-2.25; P = .001). Our results show that IPE events are associated with poor outcomes in cancer patients. Proper management plans similar to those of symptomatic pulmonary embolisms are essential.


Incorporating time-delays in S-System model for reverse engineering genetic networks.
In any gene regulatory network (GRN), the complex interactions occurring amongst transcription factors and target genes can be either instantaneous or time-delayed. However, many existing modeling approaches currently applied for inferring GRNs are unable to represent both these interactions simultaneously. As a result, all these approaches cannot detect important interactions of the other type. S-System model, a differential equation based approach which has been increasingly applied for modeling GRNs, also suffers from this limitation. In fact, all S-System based existing modeling approaches have been designed to capture only instantaneous interactions, and are unable to infer time-delayed interactions. In this paper, we propose a novel Time-Delayed S-System (TDSS) model which uses a set of delay differential equations to represent the system dynamics. The ability to incorporate time-delay parameters in the proposed S-System model enables simultaneous modeling of both instantaneous and time-delayed interactions. Furthermore, the delay parameters are not limited to just positive integer values (corresponding to time stamps in the data), but can also take fractional values. Moreover, we also propose a new criterion for model evaluation exploiting the sparse and scale-free nature of GRNs to effectively narrow down the search space, which not only reduces the computation time significantly but also improves model accuracy. The evaluation criterion systematically adapts the max-min in-degrees and also systematically balances the effect of network accuracy and complexity during optimization. The four well-known performance measures applied to the experimental studies on synthetic networks with various time-delayed regulations clearly demonstrate that the proposed method can capture both instantaneous and delayed interactions correctly with high precision. The experiments carried out on two well-known real-life networks, namely IRMA and SOS DNA repair network in Escherichia coli show a significant improvement compared with other state-of-the-art approaches for GRN modeling.


Conversion of citrate synthase into citryl-CoA lyase as a result of mutation of the active-site aspartic acid residue to glutamic acid.
The active-site aspartic acid residue, Asp-362, of Escherichia coli citrate synthase was changed by site-directed mutagenesis to Glu-362, Asn-362 or Gly-362. Only very low catalytic activity could be detected with the Asp----Asn and Asp----Gly mutations. The Asp----Glu mutation produced an enzyme that expressed about 0.8% of the overall catalytic rate, and the hydrolysis step in the reaction, monitored as citryl-CoA hydrolysis, was inhibited to a similar extent. However, the condensation reaction, measured in the reverse direction as citryl-CoA cleavage to oxaloacetate and acetyl-CoA, was not affected by the mutation, and this citryl-CoA lyase activity was the major catalytic activity of the mutant enzyme. This high condensation activity in an enzyme in which the subsequent hydrolysis step was about 98% inhibited permitted considerable exchange of the methyl protons of acetyl-CoA during catalysis by the mutant enzyme. The Km for oxaloacetate was not significantly altered in the D362E mutant enzyme, whereas the Km for acetyl-CoA was about 5 times lower. A mechanism is proposed in which Asp-362 is involved in the hydrolysis reaction of this enzyme, and not as a base in the deprotonation of acetyl-CoA as recently suggested by others. [Karpusas, Branchaud & Remington (1990) Biochemistry 29, 2213-2219; Alter, Casazza, Zhi, Nemeth, Srere & Evans, (1990) Biochemistry 29, 7557-7563].


Specific role of mitochondrial electron transport in blood-stage Plasmodium falciparum.
The origin of all mitochondria can be traced to the symbiotic arrangement that resulted in the emergence of eukaryotes in a world that was exclusively populated by prokaryotes. This arrangement, however, has been in continuous genetic flux: the varying degrees of gene loss and transfer from the mitochondrial genome in different eukaryotic lineages seem to signify an ongoing 'conflict' between the host and the symbiont. Eukaryotic parasites belonging to the phylum Apicomplexa provide an excellent example to support this view. These organisms contain the smallest mitochondrial genomes known, with an organization that differs among various genera; one genus, Cryptosporidium, seems to have lost the entire mitochondrial genome. Here we show that erythrocytic stages of the human malaria parasite Plasmodium falciparum seem to maintain an active mitochondrial electron transport chain to serve just one metabolic function: regeneration of ubiquinone required as the electron acceptor for dihydroorotate dehydrogenase, an essential enzyme for pyrimidine biosynthesis. Transgenic P. falciparum parasites expressing Saccharomyces cerevisiae dihydroorotate dehydrogenase, which does not require ubiquinone as an electron acceptor, were completely resistant to inhibitors of mitochondrial electron transport. Maintenance of mitochondrial membrane potential, however, was essential in these parasites, as indicated by their hypersensitivity to proguanil, a drug that collapsed the membrane potential in the presence of electron transport inhibitors. Thus, acquisition of just one enzyme can render mitochondrial electron transport nonessential in erythrocytic stages of P. falciparum.


A change in rabies post-exposure treatment guidelines after decision analysis in Israel.
Rabies remains a significant public health problem in Israel. Some 16 000-20 000 persons come yearly to the district health offices after being bitten by animals and 16-18% receive rabies post-exposure treatment. A quality assessment of the rabies post-exposure prophylaxis decisions was never held in Israel before. The purpose of this study was to analyse the decision-making process and to update our rabies prevention guidelines accordingly. A retrospective evaluation study of physicians' compliance with rabies post-exposure treatment guidelines in Haifa District Health Office, Ministry of Health, during 1/11/1999 to 31/12/2002. Records of all persons seeking advice (5037) following exposure to animals during the 3 years of the study were examined. Of these 2477 were eligible for the quality audit. The remaining files were analysed for the relative contribution of the different variables considered in the decision-making process. Observed agreement rate and Kappa coefficient were 0.984 and 0.803, respectively. The probability of recommending vaccination was greater when the exposure was from stray dogs as opposed to cats (OR = 48.9; 95% CI 17.9-133.3), if the wound was a bite rather than a scratch (OR = 29.0; 95% CI 19.5-43.2) or in a location defined as rabies enzootic as opposed to rabies free (OR = 7.6; 95% CI 5.8-10.1). The study demonstrated high compliance with the written guidelines. We weighted the relative importance of the variables determining the decisions. This information was used in formulating the updated guidelines.


The effects of cannabinoids on glioblastoma growth: A systematic review with meta-analysis of animal model studies.
Glioblastoma multiforme (GBM) is the most frequent and aggressive malignant brain tumour, with a poor prognosis despite available surgical and radio-chemotherapy, rising the necessity for searching alternative therapies. Several preclinical studies evaluating the efficacy of cannabinoids in animal models of GBM have been described, but the diversity of experimental conditions and of outcomes hindered definitive conclusions about cannabinoids efficacy. A search in different databases (Pubmed, Web of Science, Scopus and SciELO) was conducted during June 2019 to systematically identify publications evaluating the effects of cannabinoids in murine xenografts models of GBM. The tumour volume and number of animals were extracted, and a random effects meta-analysis of these results was performed to estimate the efficacy of cannabinoids. The impact of different experimental factors and publication bias on the efficacy of cannabinoids was also assessed. Nine publications, which satisfied the inclusion criteria, were identified and subdivided in 22 studies involving 301 animals. Overall, cannabinoid therapy reduced the fold of increase in tumour volume in animal models of GBM, when compared with untreated controls. The overall weighted standardized difference in means (WSDM) for the effect of cannabinoids was -1.399 (95% CI: -1.900 to -0.898; P-value<0.0001). Furthermore, treatment efficacy was observed for different types of cannabinoids, alone or in combination, and for different treatment durations. Cannabinoid therapy was still effective after correcting for publication bias. The results indicate that cannabinoids reduce the tumour growth in animal models of GBM, even after accounting for publication bias.


Prevalence of carbapenemases among Gram-negative bacteria in Tunisia: first report of KPC-2 producing Acinetobacter baumannii.
The rapid evolution of the antibacterial resistance problem worldwide, including the Mediterranean countries, constitutes a real threat to public health. This study aims to characterize carbapenemase encoding genes among Gram-negative bacteria collected from some Tunisian hospitals. Twenty-two clinical carbapenem-resistant Gram-negative bacteria were recovered, and identified by the matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) method. Antibiotic resistance was tested by disk diffusion method on Muller-Hinton Agar. The minimum inhibitory concentration (MIC) for imipenem was revealed by the E-test method. Carbapenemase encoding genes were screened by polymerase chain reaction (PCR). Genetic relatedness was performed by the pulsed field gel electrophoresis (PFGE) method. Our isolates, identified as K. pneumoniae (n = 7), P. mirabilis (n = 1), A. baumannii (n = 13), and P. aeruginosa (n = 1), presented high MIC values for imipenem. Enterobacerales were resistant to carbapenems due to OXA-48 production. Only, four K. pneumoniae harbored the blaNDM-1 gene. VIM-2 production was detected in P. aeruginosa. However, OXA-23 production was observed in A. baumannii isolates, one of which co-produced the KPC-2 enzyme that was identified for the first time in Tunisia in this species. A high genetic diversity was demonstrated by pulsed-field gel electrophoresis in K. pneumoniae and A. baumannii after XbaI and ApaI digestion respectively. Our findings highlight the spread of various unrelated clones of carbapenemase-producers in some Tunisian hospitals as well as the spread of several carbapenemase types.


Orthognathic Surgical Outcomes in Patients With and Without Craniofacial Anomalies.
The objective of this study is to examine hospitalization outcomes after orthognathic surgery. This study tests the hypothesis that patients with craniofacial anomalies have higher billed hospital charges, longer lengths of stay, and increased odds of development of infectious complications when compared with patients without craniofacial anomalies. The Nationwide Inpatient Sample for the years 2012 and 2013 was used. All patients who underwent an orthognathic surgical procedure were selected. The primary independent variable of interest was presence of a congenital cleft and/or craniofacial anomaly. The outcome variables were the occurrence of complications, billed hospital charges, and length of stay. Multivariable logistic and linear regression models were used to examine the effect of the presence of craniofacial anomalies on outcomes. During the study period, a total of 16,515 patients underwent an orthognathic surgical procedure in the United States. Of these patients, 2,760 had a cleft and/or craniofacial anomaly. An infectious complication occurred in 7.4% of those with a craniofacial anomaly (compared with 0.6% of those without a craniofacial anomaly). The mean billed hospital charges in those with a craniofacial anomaly was $139,317 (compared with $56,189 in those without a craniofacial anomaly). The mean length of stay in the hospital in patients with a craniofacial anomaly was 8.8 days (compared with 1.8 days in those without a craniofacial anomaly). These differences in outcomes between patients with and patients without craniofacial anomalies were significant after we adjusted for patient- and hospital-level confounders. Patients with a craniofacial anomaly are at higher risk of development of infectious complications, have higher hospital charges, and stay in the hospital for a longer duration after orthognathic surgery when compared with those without a craniofacial anomaly.


[Status and distribution of atmospheric ultrafine particles during Olympics Games in Beijing].
To understand the characteristics of size and concentration of the ultrafine particles (UFPs) during the 2008 Olympics in Beijing, as well as the influencing factors for number concentration of UFPs. Ultrafine particle monitor model 3031, manufactured by USA TSI Co., was used to monitor the daily number concentration of 20-500 nm particles, and descriptive analysis was conducted based on the data collected. During the Olympics, the mean value of number concentration for 20-100 nm particulate matter is 5 920.43 particles/ cm3. Particles ranging 30-50 nm account for the largest portion(23.49%) among particles of all sizes. There are two peak values on the curve of 24 hours' number concentration for 20-100 nm particles, respectively, at noon and evening. This paper indicates that the number concentration of ultrafine particles registered considerable reduction in the 2008's Olympics compared to the pre-Olympics years. And observation of the curves indicated that the number concentrations of the UFPs with bigger size are related to traffic volume, whereas for smaller size UFPs, illumination may be a more important influencing factor. Also there are significant correlations between number concentration and temperature as well as relative humidity.


Purification and partial characterization of glutamate synthase from Rhodospirillum rubrum grown under nitrogen-fixing conditions.
Glutamate synthase, a key enzyme in ammonia assimilation, has been purified from the photosynthetic bacterium Rhodospirillum rubrum. The purification procedure involves ion-exchange chromatography, affinity chromatography and gel filtration. The recovery in the procedure is high (62%) and the specific activity is 21 mumol of NADPH oxidized/min per mg. The enzyme is specific for its substrates, and no activity was demonstrated with NADH or NH4+ ions substituting for NADPH and glutamine respectively. The enzyme is composed of two dissimilar subunits with molecular masses of 53 and 152 kDa, and it is shown that Cl- ions have an effect on the aggregation of the enzyme. Km values for the substrates are: NADPH, 16 microM; 2-oxoglutarate, 10 microM; and glutamine, 65 microM. The enzyme is inhibited by amidotransferase inhibitors at micromolar concentrations. The role of the enzyme in the metabolic regulation of nitrogenase is discussed.


Screening markers for chronic atrophic gastritis in Chiapas, Mexico.
Intestinal-type gastric adenocarcinomas usually are preceded by chronic atrophic gastritis. Studies of gastric cancer prevention often rely on identification of this condition. In a clinical trial, we sought to determine the best serological screening method for chronic atrophic gastritis and compared our findings to the published literature. Test characteristics of potential screening tests (antibodies to Helicobacter pyloni or CagA, elevated gastrin, low pepsinogen, increased age) alone or in combination were examined among consecutive subjects enrolled in a study of H. pylori and preneoplastic gastric lesions in Chiapas, Mexico; 70% had chronic atrophic gastritis. English-language articles concerning screening for chronic atrophic gastritis were also reviewed. Sensitivity for chronic atrophic gastritis was highest for antibodies to H. pylori (92%) or CagA, or gastrin levels >25 ng/l (both 83%). Specificity, however, was low for these tests (18, 41, and 22%, respectively). Pepsinogen levels were highly specific but insensitive markers of chronic atrophic gastritis (for pepsinogen I <25 microg/l, sensitivity was 6% and specificity was 100%; for pepsinogen I:pepsinogen II ratio <2.5, sensitivity was 14% and specificity was 96%). Combinations of markers did not improve test characteristics. Screening test characteristics from the literature varied widely and did not consistently identify a good screening strategy. In this study, CagA antibodies alone had the best combination of test characteristics for chronic atrophic gastritis screening. However, no screening test was both highly sensitive and highly specific for chronic atrophic gastritis.


[Clinical value of polyposition quantitative 99mTc-pyrophosphate and 99mTc-sulfur colloid scintigraphy in assessing the degree of spreading of tumorous process in lymphogranulomatosis].
To assess the degree of tumor proliferation radionuclide studies were performed in 187 patients with lymphogranulomatosis. Scintigraphy was conducted with 99mTc-pyrophosphate and 99mTc-sulfur colloid on gammer chambers FO GAMMA LFOV and OH 110 subsequently followed by processing a scintigraphic image on special-purpose computers CDP 11/34 and OH DC 160. The signs of bone affection revealed by 99mTc-pyrophosphate were supported in 86% of cases roentgenotomographically and histologically. A correlation between scintigraphic findings and the results of morphological studies in 45 patients showed a coincidence of the results in 93% of the cases with scintigraphy of the liver and in 81% of cases with radionuclide examination of the spleen by 99mTc-sulfur colloid. A statistical analysis has indicated that scintigraphic findings are consistent with histological ones qualitatively, the probability value being p > 0.999 (X2 = 15.35).


IL-15 can signal via IL-15Rα, JNK, and NF-κB to drive RANTES production by myeloid cells.
IL-15 plays many important roles within the immune system. IL-15 signals in lymphocytes via trans presentation, where accessory cells such as macrophages and dendritic cells present IL-15 bound to IL-15Rα in trans to NK cells and CD8(+) memory T cells expressing IL-15/IL-2Rβ and common γ chain (γ(c)). Previously, we showed that the prophylactic delivery of IL-15 to Rag2(-/-)γ(c)(-/-) mice (mature T, B, and NK cell negative) afforded protection against a lethal HSV-2 challenge and metastasis of B16/F10 melanoma cells. In this study, we demonstrated that in vivo delivery of an adenoviral construct optimized for the secretion of human IL-15 to Rag2(-/-)γ(c)(-/-) mice resulted in significant increases in spleen size and cell number, leading us to hypothesize that IL-15 signals differently in myeloid immune cells compared with lymphocytes, for which IL-15/IL-2Rβ and γ(c) expression are essential. Furthermore, treatment with IL-15 induced RANTES production by Rag2(-/-)γ(c)(-/-) bone marrow cells, but the presence of γ(c) did not increase bone marrow cell sensitivity to IL-15. This IL-15-mediated RANTES production by Rag2(-/-)γ(c)(-/-) bone marrow cells occurred independently of the IL-15/IL-2Rβ and Jak/STAT pathways and instead required IL-15Rα signaling as well as activation of JNK and NF-κB. Importantly, we also showed that the trans presentation of IL-15 by IL-15Rα boosts IL-15-mediated IFN-γ production by NK cells but reduces IL-15-mediated RANTES production by Rag2(-/-)γ(c)(-/-) myeloid bone marrow cells. Our data clearly show that IL-15 signaling in NK cells is different from that of myeloid immune cells. Additional insights into IL-15 biology may lead to novel therapies aimed at bolstering targeted immune responses against cancer and infectious disease.


Radiotherapy and CD40 Activation Separately Augment Immunity to Checkpoint Blockade in Cancer.
Immunotherapy in pancreatic ductal adenocarcinoma (PDA) remains a difficult clinical problem despite success in other disease types with immune checkpoint blockade (ICB) and chimeric antigen receptor T-cell therapy. Mechanisms driving immunosuppression and poor T-cell infiltration in PDA are incompletely understood. Here, we use genetically engineered mouse models of PDA that recapitulate hallmarks of human disease to demonstrate that CD40 pathway activation is required for clinical response to radiotherapy and ICB with αCTLA-4 and αPD-1. The combination of an agonist αCD40 antibody, radiotherapy, and dual ICB eradicated irradiated and unirradiated (i.e., abscopal) tumors, generating long-term immunity. Response required T cells and also short-lived myeloid cells and was dependent on the long noncoding RNA myeloid regulator Morrbid Using unbiased random forest machine learning, we built unique, contextual signatures for each therapeutic component, revealing that (i) radiotherapy triggers an early proinflammatory stimulus, ablating existing intratumoral T cells and upregulating MHC class I and CD86 on antigen-presenting cells, (ii) αCD40 causes a systemic and intratumoral reorganization of the myeloid compartment, and (iii) ICB increases intratumoral T-cell infiltration and improves the CD8 T-cell:regulatory T-cell ratio. Thus, αCD40 and radiotherapy nonredundantly augment antitumor immunity in PDA, which is otherwise refractory to ICB, providing a clear rationale for clinical evaluation.Significance: Radiotherapy and αCD40 disrupt key links between innate and adaptive immunity, ameliorating resistance to immune checkpoint blockade in pancreatic cancer via multiple cellular mechanisms. Cancer Res; 78(15); 4282-91. ©2018 AACR.


Molecular Recognition with Macrocyclic Receptors for Application in Precision Medicine.
ConspectusMacrocyclic receptors can serve as alternatives to natural recognition systems as recognition tools. They provide effectively preorganized cavities to encapsulate guests via host-guest interactions, thereby affecting the physiochemical properties of the guests. Macrocyclic receptors exhibit chemical and thermal stabilities higher than those of natural receptors and thus are expected to resist degradation inside the body. This reduces the risk of harmful degradation byproducts and ensures optimal levels of effectiveness. Macrocyclic receptors have precise molecular weights and well-defined structures; this ensures their batch-to-batch reproducibility, which is critical for ensuring quality and effectiveness levels. Moreover, macrocyclic receptors exhibit broad modification tunabilities, rendering them adaptable to various guests. Molecular recognition is the basis of numerous biological processes. Macrocyclic receptors may display considerable potential for application in diagnosing and treating diseases, depending on the host-guest recognition of bioactive molecules. However, the binding affinities and selectivities of macrocyclic receptors toward bioactive molecules are generally insufficient, which may lead to problems such as low diagnosis accuracies, off-target leaking, and interference with normal functions. Therefore, addressing the challenge of the strong and specific complexation of bioactive molecules and macrocyclic receptors is imperative.To overcome this challenge, we proposed the innovative strategies of longitudinal cavity extension and coassembled heteromultivalent recognition for application in the recognition of small molecules and biomacromolecules, respectively. The deepened cavity provides a stronger hydrophobic effect and a larger interaction area while maintaining the framework rigidity. By coassembling two macrocyclic amphiphiles into one ensemble, we achieved the desired heteromultivalent recognition. This strategy affords the necessary binding properties while preventing the requirement of tedious steps and site mismatch in covalent synthesis. Using these two strategies, we achieved specific and strong binding of macrocyclic receptors to various bioactive molecules including biomarkers, drugs, and disease-related peptides/proteins. We then applied these macrocyclic receptor-based recognition systems in biosensing and bioimaging, drug delivery, and therapeutics.In this Account, we summarize the strategies we used in the recognition of small molecules and biomacromolecules. Thereafter, we discuss their applications in precision medicine, involving the (1) sensing of biomarkers and imaging of lesion sites, which are critical in the early screening of diseases and accurate diagnoses; (2) precise loading and targeted delivery of drugs, which are crucial in improving their therapeutic efficacies and reducing their side effects; and (3) capture and removal of disease-related biomacromolecules, which are significant for precise intervention in life processes. Finally, we propose recommendations for the further development of macrocyclic receptor-based recognition systems in biomedicine. Macrocyclic receptors exhibit considerable potential for research, and continued investigation may not only expand the applications of supramolecular chemistry but also open novel avenues for the development of precision medicine.


Lipoprotein-binding sites in human corpus luteum membrane fractions.
In a previous report evidence was presented that plasma low density lipoprotein (LDL), but not high density lipoprotein (HDL), is the major source of cholesterol used by the human corpus luteum for progesterone biosynthesis and, that plasma LDL is taken up by corpus luteum tissues via a receptor-mediated endocytotic process. Using membrane fractions prepared from fresh corpus luteum tissue obtained from nine women at various phases of the menstrual cycle, we conducted the present investigation to characterize lipoprotein-binding sites and changes in such binding sites that might occur throughout the ovarian cycle. High affinity, low capacity binding sites for [125I]iodo-LDL and also for [125I]iodo-HDL were detected in membrane fractions prepared from fresh corpus luteum tissue. The interaction of [125I]iodo-LDL with the high affinity binding sites in fresh corpus luteum membrane fractions was prevented by incubation with heparin. Also, the binding capacity of corpus luteum membrane fractions for [125I]iodo-LDL was reduced by treatment with pronase. On the other hand, the specific binding capacity for [125I]iodo-HDL was unaffected by pronase treatment. The binding capacity for [125I]iodo-LDL in membrane fractions prepared from midluteal phase corpus luteum was significantly greater than that of membrane fractions from tissue obtained at any other phase of the cycle, a finding that suggests that changes in progesterone biosynthesis throughout the cycle are positively correlated with changes in the numbers of binding sites for LDL in the corpus luteum.


[Antipsychotic drugs and hypokalemia].
The author analyzed 52 cases whose hypokalemic changes in EKG were definitely caused by antipsychotic drugs and found that the incidence was 7.6% and significantly higher in females than in males. It was suggested that the hypokalemic changes in EKG, the potassium; in serum and clinical manifestation of hypokalemia were not 1/2 parallel with each other. The EKG abnormalities were considered as the result of disturbance of intra-extracellular potassium proportion. The mechanism may be multiplex and still suggestive. It was also hinted that the sudden death occurred in patients taking APD may be related with hypokalemia. All of these provided the inspiration of further investigation.


Gold Polar Intermetallics: Structural Versatility through Exclusive Bonding Motifs.
The design of new materials with desired chemical and physical characteristics requires thorough understanding of the underlying composition-structure-property relationships and the experimental possibility of their modification through the controlled involvement of new components. From this point of view, intermetallic phases, a class of compounds formed by two or more metals, present an endless field of combinations that produce several chemical compound classes ranging from simple alloys to true ionic compounds. Polar intermetallics (PICs) belong to the class that is electronically situated in the middle, between Hume-Rothery phases and Zintl compounds and possessing e/a (valence electron per atom) values around 2. In contrast to the latter, where logical rules of formation and classification systems were developed decades ago, polar intermetallics remain a dark horse with a huge diversity of crystal structures but unclear mechanisms of their formation. Partial incorporation of structural and bonding features from both nonpolar and Zintl compounds is commonly observed here. A decent number of PICs can be described in terms of complex metallic alloys (CMAs) following the Hume-Rothery electron-counting schemes but exhibit electronic structure changes that cannot be explained by the latter. Our research is aimed at the discovery and synthesis of new polar intermetallic compounds, their structural characterization, and investigation of their properties in line with the analysis of the principles connecting all of these components. Understanding of the basic structural tendencies is one of the most anticipated outcomes of this analysis, and systematization of the available knowledge is the initial and most important step. In this Account, we focus on a well-represented but rather small section of PICs: ternary intermetallic compounds of gold with electropositive and post-transition metals of groups 12 to 15. The strong influence of relativistic effects in its chemical bonding results in special, frequently unique structural motifs, while at the same time gold participates in common structure types as an ordinary transition element. Enhanced bonding strength leads to the formation and stabilization of complex homo- and heteroatomic clusters and networks that are compositionally restricted to just a few options throughout the periodic table. Because it has the highest absolute electronegativity among metals, comparable to those of some halogens, gold usually plays the role of an anion, even being able to form true salts with the most electropositive metals. We discuss the occurrence of the structure types and show the place of gold intermetallics in the general picture. Among the structures considered are ones as common as AlB2 or BaAl4 types, in line with the recently discovered diamond-like homoatomic metal networks, formation of local fivefold symmetry, different types of tunneled structures, and more complex intergrown multicomponent structures.


Cytokine and adhesion molecule expression evolves between the neutrophilic and lymphocytic phases of viral meningitis.
Viral meningitis is characterized by cerebrospinal fluid (CSF) lymphocyte pleocytosis, although neutrophils may predominate in the early phase. The T helper 1 (Th1)/Th2 cytokine balance and expression of adhesion molecules seem to be involved in the CSF chemotaxis. We aimed to determine expression of cytokines and adhesion molecules in enteroviral meningitis. We investigated the serum and CSF levels of adhesion molecules (E-selectin, L-selectin, vascular cell adhesion molecule-1 [VCAM-1], and intracellular adhesion molecule-1 [ICAM-1]) and cytokines (interleukin-12 [IL-12] and IL-4) in 105 children during an outbreak of enteroviral meningitis. Diagnosis was confirmed with positive polymerase chain reaction (PCR) and/or serology for echovirus or Coxsackie virus, and matched with control subjects for clinical features but with negative PCR and/or serology. Apart from VCAM-1, the CSF levels of all investigated inflammatory molecules were significantly increased. In serum, sL-selectin and ICAM-1 levels were significantly higher than control subjects. Serum and CSF L-selectin, serum VCAM-1, and CSF IL-12 were all observed to be expressed in significantly higher levels in the neutrophil-dominant subgroup (72% had duration of symptoms <24 h) than in the lymphocyte-dominant group (87.5% had duration of symptoms >24 h). Serum and CSF ICAM-1 was found at significantly higher levels in the latter group. Evolving expression of adhesion molecules and cytokines indicates a shift from Th1 to Th2 immune responses as infection progresses.


Stretching Training Rehabilitation Has Potential to Alleviate Ankylosing Spondylitis in Mice by Inactivating the Wnt/β-Catenin Pathway.
Ankylosing spondylitis (AS) is a chronic inflammatory disease characterized by invasion of the joints of the central axis that involves soft tissues and joints surrounding the spine. Stretching training rehabilitation (STR) has been widely applied for the treatment of AS. The Wnt/β-catenin signalling pathway is closely related to AS. In this study, we aimed to explore the potential molecular mechanisms underlying the protective effect of STR on AS both in vitro and in vivo. Male DBA/1 mice were employed to establish an AS animal model. Hematoxylin-eosin staining showed that STR reversed pathological damages in bone tissues and the total antioxidant capacity of AS mice and increased the antioxidant capacity by upregulating superoxide dismutase and malondialdehyde expression in DBA/1 mice. The MTT, RT-qPCR, and Western blotting results further indicated that STR improved the survival rate of cells by downregulating the expression of target genes in the Wnt/β-catenin pathway and by inhibiting cell inflammation and apoptosis. In conclusion, our findings indicated that STR treatment might be an effective therapeutical strategy for AS.


Immune checkpoints in rheumatoid arthritis: progress and promise.
Rheumatoid arthritis (RA) is one of the most prevalent autoimmune inflammatory conditions, and while the mechanisms driving pathogenesis are yet to be completely elucidated, self-reactive T cells and immune checkpoint pathways have a clear role. In this review, we provide an overview of the importance of checkpoint pathways in the T cell response and describe the involvement of these in RA development and progression. We discuss the relationship between immune checkpoint therapy in cancer and autoimmune adverse events, draw parallels with the involvement of immune checkpoints in RA pathobiology, summarise emerging research into some of the lesser-known pathways, and the potential of targeting checkpoint-related pathways in future treatment approaches to RA management.


Homozygous myotonic dystrophy: clinical and molecular studies of three unrelated cases.
We report the clinical and molecular study of three unrelated homozygous myotonic dystrophy patients. In the first family, the homozygous patient shows the classical form of the disease with two DM alleles of very different expansion sizes (1000 and 60 repeats). In the second family, the homozygous patient is mildly affected and carries a minimally expanded allele (64 repeats) and a "normal" allele (38 repeats) that increases in size when transmitted. Such an intergenerational expansion of an allele in this range of repeats has not been reported to date. The third homozygous case has late onset bilateral cataracts as the only symptom. She has two minimally expanded alleles (51 and 120 repeats) that showed different intergenerational enlargement during transmission to the next generation.


Antinociceptive profiles of mu and kappa opioid agonists in a rat tooth pulp stimulation procedure.
The purpose of the study was to develop a tooth pulp stimulation procedure in the awake, freely moving rat and to then quantitatively assess the analgesic effects of compounds reported to act on mu and/or kappa receptors. The mu receptor agonists produce a biphasic (primary and secondary slope) dose-response curve (DRC) whereas kappa agonist and mixed agonist/antagonist analgesics produce single-slope DRCs. The primary slopes calculated from the mu agonist biphasic DRC are steeper than the slopes calculated for the other types of opioid analgesics. The rank order of analgesic potency for the mu agonist analgesics is oxymorphone greater than morphine = methadone greater than meperidine. The rank order analgesic potency for the kappa agonist analgesics is tifluadom greater than ethylketocyclazocine greater than U50488H and for the mixed agonist/antagonist analgesics, butorphanol greater than nalbuphine greater than pentazocine. The nonsteroidal anti-inflammatory drugs, aspirin and zomepirac, are also effective analgesics in this test procedure; but the DRC slopes for these compounds are lower than all opioid analgesics tested. The opioid antagonist naloxone produces no significant changes in threshold responses. Naloxone did reverse the threshold increases produced by morphine and ethylketocyclazocine but not aspirin. This study demonstrates that the electrical stimulation of the tooth pulp in the rat can be used as an assay for evaluation of opioid and nonopioid analgesics. When minimal effective dose values of each analgesic are plotted as a function of the clinical analgesic dose a high correlation is observed.


Optical Characteristics and Chemical Composition of Dissolved Organic Matter (DOM) from Riparian Soil by Using Excitation-Emission Matrix (EEM) Fluorescence Spectroscopy and Mass Spectrometry.
Understanding the quantity and quality of soil dissolved organic matter (DOM) in riparian buffer zones is critical for explaining the biogeochemical processes of soil DOM in river ecosystems. Here, we investigated the dissolved organic carbon, fluorescent DOM (FDOM), and DOM molecules from riparian soils on Chongming Island in eastern China. Simultaneously, the soil DOM was extensively characterized in terms of the total aromaticity index (TAI) and several optical indices. The excitation (Ex)-emission (Em) matrix parallel factor analysis results showed that two humic-like components were present (Ex/Em = 283(364)/454 nm; 337/410 nm), a fulvic-like component (Ex/Em = 241/426 nm) and a microbial degradation component (Ex/Em = 295/382 nm). The humic-like and fulvic-like substances were the main components in the riparian soil FDOM, accounting for ~90% of the FDOM. Mass spectrometry provided more detailed information for the soil DOM molecules. Six chemical fractions, amino acids, carbonyl compounds, fatty acids, lipids, proteins and sugars, were identified using liquid chromatography with quadrupole time-of-flight mass spectrometry. Lipids, proteins, and carbonyl compounds were dominant in the soil DOM, accounting for >85% of the detected molecules (m/z < 1000). Significant differences were observed between the quantities of the six soil DOM chemical fractions at the different sampling locations. Discriminant molecules verified the hypothesis that the chemical soil DOM fractions varied with the land use of the adjacent watersheds. The TAI for the soil DOM could provide more reliable information regarding the biogeochemical processes of DOM. The carbonyl compounds and lipid fractions controlled this index. Overall, the optical indices and TAI values can improve our understanding of soil DOM quality; however, the optical indices did not provide quantitative evidence regarding the sources or properties of the soil DOM. The observations from this study provided detailed information regarding the soil DOM quality and the presence of specific molecules and improved our understanding of the biogeochemical processes of DOM.


The effect of birth weight on lung spirometry in white, school-aged children and adolescents born at term: a longitudinal population based observational cohort study.
To evaluate how birth weight affects lung function measurements in childhood and adolescence in term-born children. We used data for white, term-born, singletons, from the Avon Longitudinal Study of Parents and Children to determine the association between birth weight and lung function at age 8-9 (n=4086) and 14-17 (n=2582) years. z-scores for lung function measures, adjusted for sex, height, and age, were modeled in terms of birth weight z-score adjusted for sex. In addition, gestation and head circumference then confounders (maternal smoking during pregnancy and social class) were added to the model. At age 8-9 years, birth weight z-scores were significantly associated with lung function z-scores (forced expiratory volume in 1 second, forced vital capacity [FVC], and forced mid-expiratory flow between 25% and 75% of FVC). These relationships essentially were unchanged when birth weight z-scores were further adjusted for gestation, head circumference, and confounders, except for forced mid-expiratory flow between 25% and 75% of FVC, which was no longer significant after we adjusted for head circumference and confounders. At age 14-17 years, the associations between adjusted birth-weight z-scores and spirometry z-scores were in general not significant. Estimated differences for forced expiratory volume in 1 second were 30 mL at ages 8-9 years and 33 mL at 14-17 years for 1 kg change in birth-weight standardized for gestation and sex. Birth weight is associated with lung function in term-born children at 8-9 years, but less so at 14-17 years, suggesting that birth weight influences lung function in early childhood but has lesser effect later in life.


Focal glomerulosclerosis in renal allografts: association with the nephrotic syndrome and chronic rejection.
Focal segmental and/or global sclerotic glomerular lesions with hyalinosis were noted in three allografts in association with the nephrotic syndrome (NS) and chronic rejection (CR). Similar lesions were absent in eight allografts with CR without NS. Previous reports have stressed the presence of this lesion in allografts with recurrence of the disease entity "idiopathic nephrotic syndrome with focal glomerulosclerosis". Both clinical and pathologic evidence suggest that recurrence was not a factor in the pathogenesis of the lesion in the three allografts with CR and NS. The presence of these lesions in failing allografts may represent the result of CR with associated NS rather than recurrence of the disease entity.


Height as a risk factor for osteosarcoma.
Previous investigations have suggested that osteosarcoma may be associated with a taller stature, but the relationship between height and osteosarcoma remains controversial. Height at diagnosis was evaluated in a continuous series of 962 osteosarcoma subjects treated between 1981 and 2001. Patients diagnosed during growth (group 1) were separated from those diagnosed in adulthood (group 2). Height (H) and final height (FH) were expressed as standard deviation scores (SDS), calculated by national reference data. Group 1 subjects were above the 50th centile and their mean H-SDS values (0.31 +/- 1.1) were significantly higher than the mean FH-SDS values (P < 0.0001) of the group 2 subjects, both in males and females. In contrast, the mean FH-SDS (0.01 +/- 1.1) of group 2 did not differ from that of the reference population. The highest incidence of osteosarcoma was at 12.5 years in females, 14.5 years in males. These data confirm previous observations of an association between osteosarcoma development and height, at least in growing individuals. The higher incidence during the pubertal spurt, in the anatomic sites of greater growth and in taller individuals, suggests that growth factors play an important role in the pathogenesis of this bone cancer.