Stromal fibrosis: imaging features with diagnostic contribution of diffusion-weighted MRI.
To describe magnetic resonance imaging (MRI) and ultrasonography findings of breast stromal fibrosis (SF) and compare apparent diffusion coefficient (ADC) stromal fibrosis values with breast cancer and normal parenchyma. 45 patients (ages 22‒74) with histopathologically proven SF who underwent MRI were included in this study. Their MRI and ultrasonography features were examined and categorized. The mean ADC values for SF, contralateral normal parenchyma, and breast malignancy of the control group values were calculated and compared among each other. The vast majority of SF on sonography showed features suggestive of malignancy: (1) irregular in shape 25/45 (55%); (2) indistinct in margin 27/45 (60%); and (3) hypoechoic 39/45 (87%) with posterior acoustic shadowing 11/45 (24%). An SF MRI showed a mass in 12/45 (26%) and non-mass enhancement in 33/45 (74%), mostly with irregular (8/12; 67%) shape. Non-mass lesions showed heterogeneous (12/33), clumped (9/33), and homogenous (9/33) enhancement. The initial SF contrast uptake rate varied between slow (57%), rapid (22%), and medium (21%). Delayed SF enhancement may be persistent (66%) or plateau (34%). Small cysts were located around/near 21 (47%) of lesions. Ductal ectasia was found in 14 (31%) of all patients. Mean ADCs of parenchyma, SF, and malignancy were 1.32 ± 0.32, 1.23 ± 0.25, and 0.99 ± 0.24 × 10-3 mm2 sec-1, respectively. SF often mimics breast carcinoma on imaging and leads the radiology‒pathology disagreement. In terms of distinguishing SF from malignancy, ADC could be a significant and promising value in diffusion-weighted MRI along with conventional sequences. Slow initial uptake with delayed persistent contrast enhancement in a non-mass lesion with relatively higher ADC values are very helpful for differentiating SF from malignancy. The presence of small cysts and ductal ectasia were common findings around/near the SF. Advances in knowledge: A quantitative analysis for measuring ADC values along with additional MRI features can be very helpful in distinguishing SF from malignant lesions.


Exploring novel KDR inhibitors based on pharmaco-informatics methodology.
Kinase-insert domain-containing receptor (KDR) is one of the important mediators of Vascular endothelial growth factor (VEGF) function in endothelial cells. Inhibition of KDR can be therapeutically advantageous for treatment of a number of diseases. The present study focuses on exploring novel KDR inhibitors by means of pharmaco-informatics methodologies. Three-dimensional quantitative structure-activity relationship (3D-QSAR) analysis by atom-based pharmacophore mapping over a set of 85 molecules provides a proposition regarding the molecular fingerprint that can be optimized for designing more active inhibitors. The model was statistically validated with Q(2) = 0.865 for training and r(2) = 0.789, Pearson-r = 0.903 for test set molecules; r(2)(0.925) by external validation suggests model robustness and indicates it as a strong query for screening any compound library. Virtual screening shows the importance of active site and hinge region residue for interaction with KDR inhibitors. Remarkably the retrieved hits contain a urea backbone, implicating urea derivatives as promising candidate for designing KDR inhibitors. The hydrophobicity of active site, which has until now been overlooked, has been raised into the picture by this study. This can impact on KDR drug development. The study thus quantifies crucial structural requirements necessary for a favourable interaction with the receptor binding site while the cooperative pattern provides important structural clues to chemists for framing potent medicinal agents in future.


Generation of phase-shifting algorithms with N arbitrarily spaced phase-steps.
Phase-shifting (PS) is an important technique for phase retrieval in interferometry (and three-dimensional profiling by fringe projection) that requires a series of intensity measurements with known phase-steps. Usual PS algorithms are based on the assumption that the phase-steps are evenly spaced. In practice, however, this assumption is often not satisfied exactly, which leads to errors in the recovered phase. In this work we present a systematic algebraic approach for generating general PS algorithms with N arbitrarily spaced phase-steps, which present advantages (e.g., the PS error can be avoided) over known algorithms that assume equally spaced phase-steps. Simulations are presented.


Molecular mechanisms of cellular proliferation in acute myelogenous leukemia by leptin.
Leptin acts as a growth factor in normal cells as well as in various types of cancer cells. We investigated the effects of leptin on human acute myelogenous leukemia (AML) cells. Leptin stimulated the proliferation of HEL cells through the phosphorylation of STAT3 and ERK1/2. The blocking of STAT3 phosphorylation with the specific inhibitor, AG490, significantly reduced leptin-induced ERK1/2 phosphorylation and cellular proliferation, whereas the blocking of ERK1/2 activation by the specific ERK1/2 inhibitor, PD98059, did not affect the STAT3 phosphorylation or leptin-induced proliferation in HEL cells. Furthermore, knockdown of leptin receptor (OB-R) expression with stealth RNA interference (RNAi) reduced the leptin-induced proliferation of HEL cells and also significantly attenuated leptin-induced STAT3 and ERK1/2 activation. These results suggest that leptin promotes AML cell growth by activating STAT3 and MAPK, although not directly dependent on ERK.


Multifunctional molecule ERp57: From cancer to neurodegenerative diseases.
The protein disulfide isomerase (PDI) gene family is a protein family classically characterized by endoplasmic reticulum (ER) localization and isomerase and redox activity. ERp57, a prominent multifunctional member of the PDI family, is detected at various levels in multiple cellular localizations outside of the ER. ERp57 has been functionally linked to a host of physiological processes and numerous studies have demonstrated altered expression and aberrant functionality of ERp57 in association with diverse pathological states. Here, we summarize available knowledge of ERp57's functions in subcellular compartments and the roles of dysregulated ERp57 in various diseases toward an emphasis on the potential utility of therapeutic development of ERp57.


Numerical Investigation of the Microscopic Heat Current Inside a Nanofluid System Based on Molecular Dynamics Simulation and Wavelet Analysis.
Molecular dynamics simulation is employed to investigate the microscopic heat current inside an argon-copper nanofluid. Wavelet analysis of the microscopic heat current inside the nanofluid system is conducted. The signal of the microscopic heat current is decomposed into two parts: one is the approximation part; the other is the detail part. The approximation part is associated with the low-frequency part of the signal, and the detail part is associated with the high-frequency part of the signal. Both the probability distributions of the high-frequency and the low-frequency parts of the signals demonstrate Gaussian-like characteristics. The curves fit to data of the probability distribution of the microscopic heat current are established, and the parameters including the mean value and the standard deviation in the mathematical formulas of the curves show dramatic changes for the cases before and after adding copper nanoparticles into the argon base fluid.


The human complement inhibitor Sushi Domain-Containing Protein 4 (SUSD4) expression in tumor cells and infiltrating T cells is associated with better prognosis of breast cancer patients.
The human Sushi Domain-Containing Protein 4 (SUSD4) was recently shown to function as a novel inhibitor of the complement system, but its role in tumor progression is unknown. Using immunohistochemistry and quantitative PCR, we investigated SUSD4 expression in breast cancer tissue samples from two cohorts. The effect of SUSD4 expression on cell migration and invasion was studied in vitro using two human breast cancer cell lines overexpressing SUSD4. Tissue stainings revealed that both tumor cells and tumor-infiltrating cells expressed SUSD4. The highest SUSD4 expression was detected in differentiated tumors with decreased rate of metastasis, and SUSD4 expression was associated with improved survival of the patients. Moreover, forced SUSD4 expression in human breast cancer cells attenuated their migratory and invasive traits in culture. SUSD4 expression also inhibited colony formation of human breast cancer cells cultured on carcinoma-associated fibroblasts. Furthermore, large numbers of SUSD4-expressing T cells in the tumor stroma associated with better overall survival of the breast cancer patients. Our findings indicate that SUSD4 expression in both breast cancer cells and T cells infiltrating the tumor-associated stroma is useful to predict better prognosis of breast cancer patients.


CONSPIRATORIAL WHISPERS AND CONSPICUOUS DISPLAYS: GAMES OF SIGNAL DETECTION.
Recent models of signaling have assumed that the expenditure required to ensure detection of a display is negligible and have concentrated instead on the costs that may be necessary to maintain honesty. Such models predict that individuals who share the same interests are likely to communicate using "conspiratorial whispers," signals that are cheap and inconspicuous. Here, I present a game-theoretical model of signal detection (in a noisy environment, in the presence of potential eavesdroppers), which demonstrates that the idea of conspiratorial whispers is far too simplistic. It is true that in "cooperative" signaling systems (where signalers attempt to elicit responses that are beneficial for receivers), signal cost is not required to maintain honesty. However, some level of expenditure is still needed to ensure that a signal is reliably detected. Moreover, there exists a conflict of interest between signalers and receivers over the division of this expenditure. To predict the stable level of display in such cases, one needs to know how this conflict of interest will be resolved. The model reveals that the outcome may range from a whisper to a conspicuous and costly (though still conspiratorial) display. The more closely related the receiver is to the signaler, the greater the level of signal exaggeration that is expected-the opposite prediction to that of honest signaling models.


Predictability in diabetic nephropathy.
Possible association between several variables and the prognosis of nephropathy was investigated in 23 insulin-dependent diabetics with established diabetic nephropathy. Age at onset and duration of diabetes, blood pressure and metabolic control were among these variables. In the mean observation period of 27 months, end-stage renal disease (ESRD) developed in 11 patients. Nine patients died, including six with progression to ESRD, mainly of cardiac disease. Age, duration of diabetes, insulin requirement, blood pressure and glycosylated hemoglobin concentration showed higher values in the patients with progression to ESRD, but no difference was statistically significant. During the observation time, however, blood pressure was less well controlled in the patients who developed ESRD. The deterioration rate of renal function showed wide interindividual variation. In each patient, however, the decline seemed to follow a linear regression course. The results suggest that, in the absence of dependable prognostic factors, regular individual monitoring of glomerular filtration rate can provide a reliable predictive estimate of the progression rate.


[Investigation of biofilm formation properties of staphylococcus isolates].
Biofilm production is an important virulence factor which allows staphylococci to adhere to medical devices. The principal component of biofilm is a "polysaccharide intercellular adhesin (PIA)" which is composed of a beta-1,6-N-acetylglucosamine polymer synthesized by an enzyme (N-acetylglucosamine transferase) encoded by the ica operon found on the bacterial chromosome. This operon is composed of four genes (A, B, C, and D), and a transposable element IS256. In this study, we aimed to determine the biofilm production characteristics of invasive/non-invasive staphylococcus isolates and different staphylococcus species. Biofilm production of 166 staphylococci was phenotypically investigated on Congo Red Agar (CRA); the presence of icaA, icaD and IS256 genes were investigated by polymerase chain reaction (PCR). 74 of the isolates (44.6%) were identified as methicillin resistant Staphylococcus aureus (MRSA), 25 (15.1%) as methicillin sensitive S.aureus (MSSA), 25 (37.3%) as Staphylococcus hominis, 20 (12%) as S.epidermidis, ten (15%) as Staphylococcus haemolyticus, nine (13.4%) as Staphylococcus capitis, two (3%) Staphylococcus saprophyticus and one (1.5%) as Staphylococcus warnerii. Of the MRSA strains, 52 were isolated from blood and 22 from nose; all MSSA strains were isolated from nose cultures. Coagulase-negative staphylococci (CoNS) strains were composed of invasive and non-invasive strains isolated from nose, catheter tip and blood cultures from patients with catheter. Production with CRA method was found to be statistically significant in invasive isolates (p< 0.001). It is concluded that; as the biofilm formation capacity of invasive isolates can cause refractory infections and the importance of carriage and hospital infections of these bacteria, it is important to prevent the spread of these isolates. A combination of phenotypic and genotypic tests is recommended for the investigation of biofilm formation in staphylococci. 40.3% of the CoNS isolates, and 85.8% of S.aureus isolates produced biofilm on CRA (p< 0.001) and with PCR method the ratio of carrying three genes was found to be statistically important in S.aureus when compared with CoNS. Carriage of three genes and biofilm formation capacity of invasive isolates can cause refractory infections and the importance of carriage and hospital infections of these bacteria, it is important to prevent the spread of these isolates. A combination of phenotypic and genotypic tests is recommended for the investigation of biofilm formation in staphylococci.


Alternariol induces abnormal nuclear morphology and cell cycle arrest in murine RAW 264.7 macrophages.
The mycotoxin alternariol (AOH), a frequent contaminant in fruit and cereal products, is known to induce DNA damage with subsequent cell cycle arrest. Here we elucidated the effects of AOH on stages of cell cycle progression using the RAW 264.7 macrophage model. AOH resulted in an accumulation of cells in the G2/M-phase (4N). Most cells exhibited a large G2 nucleus whereas numbers of true mitotic cells were reduced relative to control. Both cyclin B1 and p-cdc2 levels increased, while cyclin B1 remained in the cytoplasm; suggesting arrest in the G2/M transition point. Remarkably, after exposure to AOH for 24h, most of the cells exhibited abnormally shaped nuclei, as evidenced by partly divided nuclei, nuclear blebs, polyploidy and micronuclei (MN). AOH treatment also induced abnormal Aurora B bridges, suggesting that cytokinesis was interfered within cells undergoing karyokinesis. A minor part of the resultant G1 tetraploid (4N) cells re-entered the S-phase and progressed to 8N cells.


Health benefits in 2006: premium increases moderate, enrollment in consumer-directed health plans remains modest.
Based on a survey of 2,122 randomly selected public and private employers, this paper reports on the state of employer-sponsored health insurance in spring 2006, including recent changes. Premiums increased 7.7 percent from spring 2005 to spring 2006 and have risen 68 percent since 2001. About 4 percent of workers are enrolled in high-deductible health plans with savings options. The percentage of workers covered by their own employer did not statistically change from 2005 to 2006.


Mass transfer characterization of an airlift probe for oxygenating and mixing cell suspensions in an NMR spectrometer.
The oxygen transfer characteristics of a 20-mm O.D. airlift contactor fitted with an oxygen microelectrode were determined by steady-state sulfite oxidation measurements. The volumetric mass transfer coefficient k(L)a was proportional to sparging power input per unit volume raised to a power which varied from 0.41 in water (coalescing bubbles) to 0.76 in NaCl solutions (noncoalescing bubbles). The highest observed k(L)a value was 0.012 s(-1) which is sufficient to aerate Escherichia coli in an NMR spectrometer at moderate to high cell densities, depending on the physiological state of the cells.


Retinal Vascular Changes During Pregnancy Detected With Optical Coherence Tomography Angiography.
To evaluate retinal vascular status during pregnancy by using optical coherence tomography angiography (OCTA). Women in their third trimester of pregnancy and nonpregnant age-matched women were recruited for this prospective, case-control study. Subjects were imaged with OCTA. Main outcome measures were foveal avascular zone parameters, perfusion density (PD) percentage in the superficial retinal capillary plexus (SCP), PD percentage in the deep retinal capillary plexus (DCP), SCP vessel length density (VLD), DCP-VLD, and choriocapillaris (CC) flow voids (i.e., flow deficits in the CC). Nineteen eyes of 10 pregnant subjects and 44 eyes of 27 nonpregnant control women were included. Mean ages were 36 ± 7 and 35 ± 8 years (SD), respectively (P value = 0.78). Mean gestational age of pregnant women was 33 weeks (range = 29-39, SD = 3). There was a significant reduction in the SCP-PD in the entire scan and in the nasal Early Treatment Diabetic Retinopathy Study subfield (47.9 vs. 49.7, P = 0.04 and 49.3 vs. 51.6, P = 0.03, respectively) in the pregnant cohort versus controls. There was a significant increase in the DCP-PD in the parafoveal region and in the temporal and inferior Early Treatment Diabetic Retinopathy Study subfields (58.0 vs. 55.9, P = 0.03; 57.9 vs. 55.5, P = 0.02; 58.0 vs. 55.9, P = 0.05, respectively) in the pregnant cohort. There was no significant difference in foveal avascular zone parameters, SCP-VLD, DCP-VLD, or CC flow voids between the two populations. This study detected retinal vasculature changes in the third trimester of pregnancy. Mean SCP-PD was significantly decreased and mean DCP-PD was significantly increased without a difference in VLD.


FOXN1 Duplication and Congenital Hypertrichosis.
We report a case of congenital hypertrichosis and FOXN1 duplication. FOXN1 is a member of the forkhead box gene family, located on chromosome 17. Its function includes differentiation of epithelial cells and regulation of keratinocytes, especially hair keratins. Loss of function of these transcription factors leads to a disruption in hair growth. As far as we are aware, this is the first case of FOXN1 duplication associated with congenital hypertrichosis to be reported in the literature.


In vivo three-dimensional molecular imaging with Biosensor Imaging of Redundant Deviation in Shifts (BIRDS) at high spatiotemporal resolution.
Spectroscopic signals which emanate from complexes between paramagnetic lanthanide (III) ions (e.g. Tm(3+)) and macrocyclic chelates (e.g. 1,4,7,10-tetramethyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate, or DOTMA(4-)) are sensitive to physiology (e.g. temperature). Because nonexchanging protons from these lanthanide-based macrocyclic agents have relaxation times on the order of a few milliseconds, rapid data acquisition is possible with chemical shift imaging (CSI). Thus, Biosensor Imaging of Redundant Deviation in Shifts (BIRDS) which originate from nonexchanging protons of these paramagnetic agents, but exclude water proton detection, can allow molecular imaging. Previous two-dimensional CSI experiments with such lanthanide-based macrocyclics allowed acquisition from ~12-μL voxels in rat brain within 5 min using rectangular encoding of k space. Because cubical encoding of k space in three dimensions for whole-brain coverage increases the CSI acquisition time to several tens of minutes or more, a faster CSI technique is required for BIRDS to be of practical use. Here, we demonstrate a CSI acquisition method to improve three-dimensional molecular imaging capabilities with lanthanide-based macrocyclics. Using TmDOTMA(-), we show datasets from a 20 × 20 × 20-mm(3) field of view with voxels of ~1 μL effective volume acquired within 5 min (at 11.7 T) for temperature mapping. By employing reduced spherical encoding with Gaussian weighting (RESEGAW) instead of cubical encoding of k space, a significant increase in CSI signal is obtained. In vitro and in vivo three-dimensional CSI data with TmDOTMA(-), and presumably similar lanthanide-based macrocyclics, suggest that acquisition using RESEGAW can be used for high spatiotemporal resolution molecular mapping with BIRDS.


Epithelial stem/progenitor cells in lung postnatal growth, maintenance, and repair.
The adult lung consists of a trachea leading into a system of branched airways ending in millions of alveolar sacs. It contains many different epithelial cell types arranged in precise patterns along the proximodistal axis. Each region of the lung has the capacity to repair through the proliferation of different epithelial cell types. However, the precise identity of the cells mediating repair is not fully resolved. To address this problem, we are using genetic lineage-labeling techniques in the mouse. The tools we have made will also be useful for understanding how progenitor cell behavior is regulated under normal and pathological conditions.


Dust accumulation due to anthropogenic impact induces anatomical and photochemical changes in leaves of Centranthus ruber growing on the slope of the Vesuvius volcano.
In Mediterranean ecosystems, some natural areas are exposed to severe anthropogenic impact. Especially in summer, the considerable number of tourists visiting such areas, often with vehicles, causes deposition of dust over the vegetation due to formation of powder clouds, also favoured by wind erosion, high temperature, low precipitation and incoherent soil structure. The main aim of this study was to analyse whether the deposition of dust can induce changes in leaf anatomical functional traits and in the efficiency of photosynthetic apparatus in Centranthus ruber, a species widespread in Mediterranean ecosystems. Leaf morpho-functional traits were quantified in plants growing at sites characterised by high (HD) and low (LD) dust deposition, in periods with high anthropogenic impact. Analyses included quantification of chlorophyll fluorescence emission parameters, photosynthetic pigment concentration as well as stomatal size and frequency, leaf lamina thickness, quantification of intercellular spaces and phenolics in the mesophyll through microscopy. The overall analysis suggested that the different conditions of dust deposition induced different adjustment of morpho-functional traits in leaves of C. ruber. High dust deposition shielded the leaf lamina, protecting the photosynthetic apparatus from excess light and favoured plant photochemical efficiency. Leaves exposed to low dust deposition showed higher accumulation of phenolic compounds, protecting chloroplast membranes and characterised by high thermal dissipation of excess light. Such adaptive phenomena can affect vegetation dynamics due to possible different species-specific plant responses, resulting in different plant competitiveness under the limiting conditions of Mediterranean environments.


Expanding the role of the virome: commensalism in the gut.
Viruses affect host physiology beyond causing acute disease, thereby giving rise to the concept that the virome is a component of the microbiome. However, the role of the enteric virome is understudied relative to the fast-paced research examining commensal bacteria in the intestine. In this article, I discuss our recent work on murine norovirus indicating that an animal virus in the intestine can provide many of the signals to the host that have been attributed to commensal bacteria. Our findings suggest that the surge in microbiome research should incorporate examination of the enteric virome.


The Association Between Binge-Watching Behavior and Psychological Problems Among Adolescents.
This study aims to investigate the relationship between binge-watching addiction and family, emotional, conduct, cognitive and anger control problems, hyperactivity, Diagnostic and Statistical Manual of Mental Disorders, 4th Edition (DSM-IV)-ADHD Index, DSM-IV-Inattention, and DSM-IV-Hyperactivity Impulsivity in a large sample of healthy Turkish adolescents. The association between binge-watching behavior addiction and mood disorders in healthy adolescents was examined in this cross-sectional study conducted in Turkey. Participants completed both measures, The Binge-Watching Addiction Questionnaire and The Conners-Wells' Adolescent Self-Report Scale. An online survey of 189 adolescents was conducted. One hundred fourteen subjects (60.3%) were categorized into the binge-watcher group. Results showed important positive associations between binge-watching behavior and emotional, conduct, cognitive problems, and DSM-IV-Inattention. The conducted analysis showed an important association between high frequency of binge-watching and emotional problems (p < 0.001), conduct problems (p = 0.012), cognitive problems (p = 0.001), and inattention (p < 0.001). These findings contributed to our comprehension of adolescents' psychological correlates of television viewing behavior. Further researches on the relationship between extreme binge-watching and psychiatric disorders of adolescents are recommended.


A submaximal test for the assessment of knee extensor endurance capacity.
We aimed to develop an undemanding test for endurance capacity of the knee extensor muscles, which can also be applied to frail participants. We hypothesized 1) that the first objective indications for peripheral fatigue during incremental unilateral repetitive isometric knee extensor contractions could be used to assess a fatigue threshold (FT), 2) that torque at FT would depend on training status, and 3) that this torque could easily be sustained for 30 min. Five trained and five untrained participants performed 5-min bouts of 60 repetitive contractions (3-s on and 2-s off). Torque, set at 25% maximal voluntary contraction (MVC), was increased by 5% MVC in subsequent bouts. The highest torque for which rectified surface EMG remained stable during the bout was defined as the FT. On separate occasions, 30-min bouts were performed at and above the FT to assess sustainable torque. Changes in gas exchange parameters, HR, and RPE were monitored to corroborate FT. At FT (RPE = 5.7 ± 1.7), torque was higher (P < 0.05) in trained (41.4% ± 5.8% MVC) than in untrained participants (30.5% ± 1.8% MVC). Sustainable torque was ∼4% higher than (P < 0.05) and significantly related to FT (r(2) = 0.79). When torque was increased by 5% MVC, significant increases in rectified surface EMG and V˙O2 were found. During incremental knee extensor contractions, FT could be assessed at a submaximal exercise intensity. FT was higher in trained than in untrained participants and was related to exercise sustainability. With the use of FT, changes in endurance capacity of single muscle groups can potentially also be determined in frail participants for whom exercise performed until exhaustion is unwarranted.


[New aspects of collagen formation].
New data are presented concerning the process of synthesis and secretion of collagen. The stages are described of its elaboration and biochemical transformations of collagen molecules occurring in the cell and the alterations leading to collagen fibres building-up in the extracellular spaces. Electronic microscopy of fibrogenesis study, carried out during an experimental research on lung fibrosis, has shown new aspects not yet described in the literature. It is demonstrated that collagen fibres could be formed by assembling the tropocollagen molecules in the fibroblast cytoplasm, before the expulsion from the cell. This phenomenon could represent a new way not yet mentioned among the mechanisms of collagen synthesis. It is likely that this process take place in the framework of pathological process of fibrosis development.


Application of a neural network in high-throughput protein crystallography.
High-throughput protein crystallography requires the automation of multiple steps used in the protein structure determination. One crucial step is to find and monitor the crystal quality on the basis of its diffraction pattern. It is often time-consuming to scan protein crystals when selecting a good candidate for exposure. The use of neural networks for this purpose is explored. A dynamic neural network algorithm to achieve a fast convergence and high-speed image recognition has been developed. On the test set a 96% success rate in identifying properly the quality of the crystal has been achieved.


Phosphatase UBLCP1 controls proteasome assembly.
Ubiquitin-like domain-containing C-terminal domain phosphatase 1 (UBLCP1), an FCP/SCP phosphatase family member, was identified as the first proteasome phosphatase. UBLCP1 binds to proteasome subunit Rpn1 and dephosphorylates the proteasome in vitro However, it is still unclear which proteasome subunit(s) are the bona fide substrate(s) of UBLCP1 and the precise mechanism for proteasome regulation remains elusive. Here, we show that UBLCP1 selectively binds to the 19S regulatory particle (RP) through its interaction with Rpn1, but not the 20S core particle (CP) or the 26S proteasome holoenzyme. In the RP, UBLCP1 dephosphorylates the subunit Rpt1, impairs its ATPase activity, and consequently disrupts the 26S proteasome assembly, yet it has no effects on the RP assembly from precursor complexes. The Rpn1-binding and phosphatase activities of UBLCP1 are essential for its function on Rpt1 dephosphorylation and proteasome activity both in vivo and in vitro Our study establishes the essential role of the UBLCP1/Rpn1/Rpt1 complex in regulating proteasome assembly.


Human fetal, cord blood, and adult lymphocyte progenitors have similar potential for generating B cells with a diverse immunoglobulin repertoire.
Several characteristics of the immunoglobulin (Ig) repertoire in fetuses and adults set them apart from each other. Functionally, this translates into differences in the affinity and effectiveness of the humoral immune response between adults and the very young. At least 2 possibilities could explain these differences: (1) fetal and adult lymphocyte progenitors differ significantly in their potential to form a diverse repertoire, and (2) factors extrinsic to the immunoglobulin locus are more influential to the character of the repertoire. To address this we used nonobese diabetic-severe combined immunodeficient-beta(2) microglobulin knockout (NOD/SCID/beta(2)m(-/-)) mice reconstituted with human B-cell progenitors to compare the immunoglobulin repertoire potential of human fetal, cord blood, and adult sources. We found nearly identical VH and JH gene segment use and only modest differences in the third complementarity determining region of the immunoglobulin heavy chain (HCDR3). We conclude that the repertoire potential is remarkably similar regardless of the age of the individual from which progenitors are derived. Age-related differences in the immunoglobulin repertoire and variance of B-cell responses to immunization appear to arise from selection rather than from changes in recombination of the immunoglobulin locus itself. From the standpoint of the Ig repertoire, an immune system reconstituted from fetal or neonatal stem cells would likely be as diverse as one generated from adult bone marrow.


[Testing two simple photometers (Lange) for use in small animal practice (author's transl)].
Two simple photometers were tested for their suitability for use in veterinary practice. Determinations of haemoglobin, glucose and total serum protein were studied using specially designed testing kits. The methods were found to be rapid, simple and reliable. Comparison of the results with those obtained using known standard methods showed a satisfactory correlation for haemoglobin and total serum protein. The results obtained on determination of glucose showed some minor differences which were possibly due to the different chemical methods used. Instruments as well as the methods used in testing are recommended.


CALCOCO2/NDP52 initiates selective autophagy through recruitment of ULK and TBK1 kinase complexes.
The selective macroautophagy of prospective cargo necessitates activity of the autophagy machinery at cargo-determined locations. Whether phagophore membranes are recruited to, or are generated de novo at, the cargo is unknown. In our recent study we show that damaged Salmonella-containing vacuoles, marked by LGALS8/galectin-8, engage the cargo receptor CALCOCO2/NDP52 to recruit the autophagy-initiating ULK and TBK1 complexes and cause the formation of WIPI2-positive phagophore membranes. CALCOCO2 functions in the induction of autophagy by forming a trimer with RB1CC1/FIP200 and TBKBP1/SINTBAD-AZI2/NAP1, components of the ULK and TBK1 kinase complexes, respectively. Such recruitment of the upstream autophagy machinery to prospective cargo reveals how in complex eukaryotes detection of cargo-associated 'eat me' signals, induction of autophagy, and juxtaposition of cargo and phagophores are integrated.


Hospitalisations before and after entry into a residential aged care facility: An interrupted time series analysis.
Hospitalisations are an important indicator of safety and quality of care in residential aged care facilities (RACFs). This study aimed to investigate changes in hospital use 12 months before and 12 months after RACF entry using routinely collected data from 25 Australian RACFs. This was a retrospective longitudinal cohort study using linked aged care provider and hospital record data. The sample comprised 1029 residents living in an aged care facility between July 2014 and December 2019 who had stayed a minimum of 12 months in an RACF. The outcome measures were all-cause hospitalisations and fall-related hospitalisations. We applied an interrupted time series analysis using segmented regression to examine changes in both outcome measures over time. Stratified analyses were conducted by gender and dementia status. The rate of all-cause hospitalisations increased dramatically over the 12 months before RACF entry, from 97 per 1000 residents per month 12 months prior to RACF admission to 303 per 1000 residents at the second month prior to RACF entry. All-cause hospitalisations then decreased considerably to 55 per 1000 residents upon RACF admission and stabilised across the next 12 months. Such trajectories were also observed in fall-related hospitalisations and were consistent for gender and dementia status. In this study, hospitalisation rates decreased significantly after RACF entry, and such reductions were maintained for residents who stayed for 12 months in RACFs. Multiple hospital admissions are likely to precipitate entry into RACF. Additional investigation of how community-based services can be successful in reducing the escalating hospitalisations is needed.


Rapid ELISA Using a Film-Stack Reaction Field with Micropillar Arrays.
A film-stack reaction field with a micropillar array using a motor stirrer was developed for the high sensitivity and rapid enzyme-linked immunosorbent assay (ELISA) reaction. The effects of the incubation time of a protein (30 s, 5 min, and 10 min) on the fluorescence intensity in ELISAs were investigated using a reaction field with different micropillar array dimensions (5-µm, 10-µm and 50-µm gaps between the micropillars). The difference in fluorescence intensity between the well with the reaction field of 50-µm gap for the incubation time of 30 s and the well without the reaction field with for incubation time of 10 min was 6%. The trend of the fluorescence intensity in the gap between the micro pillars in the film-stack reaction field was different between the short incubation time and the long incubation time. The theoretical analysis of the physical parameters related with the biomolecule transport indicated that the reaction efficiency defined in this study was the dominant factor determining the fluorescence intensity for the short incubation time, whereas the volumetric rate of the circulating flow through the space between films and the specific surface area were the dominant factors for the long incubation time.


Quantitative fluorescence analysis of cyclosporine binding to human leukocytes.
The purpose of this investigation was to estimate the binding of cyclosporine at the single-cell level on human peripheral lymphocytes, and to test possible identity of the cyclosporine-binding site with a common receptor of T cell activation. A dansyl-coupled derivative (Dans cyclosporine) was used as a fluorescent probe. The histograms of unseparated, labeled peripheral leukocytes obtained by a fluorescence-activated cell sorter (FACS) showed that Dans cyclosporine stained all leukocytes--but two distinct populations could be separated based on the intensity of fluorescence. The more brightly labeled cells consisted mainly of granulocytes and monocytes, whereas the less-bright cells represented the lymphocyte compartment. Fluorescence microscopy revealed binding on the membrane for both cell populations; the label was, however, rapidly internalized in phagocytes. For both populations binding was saturable, time and temperature dependent, and reversible. Half-saturation occurred at approximately 5 X 10(-7) M (Kd). With respect to lymphocyte subpopulations, no difference of cellular fluorescence was found between unseparated lymphocytes and T cell subsets. In addition, mitogens such as concanavalin A, phytohemagglutinin, phorbol 12-myristate 13-acetate, or OKT3 antibody did not inhibit Dans cyclosporine binding. These results clearly indicate that cyclosporine binds to all peripheral blood lymphocytes, and no preferential binding on T cell subsets can be detected.


Population Kinetics of 0.9% Saline Distribution in Hemorrhaged Awake and Isoflurane-anesthetized Volunteers.
Population-based, pharmacokinetic modeling can be used to describe variability in fluid distribution and dilution between individuals and across populations. The authors hypothesized that dilution produced by crystalloid infusion after hemorrhage would be larger in anesthetized than in awake subjects and that population kinetic modeling would identify differences in covariates. Twelve healthy volunteers, seven females and five males, mean age 28 ± 4.3 yr, underwent a randomized crossover study. Each subject participated in two separate sessions, separated by four weeks, in which they were assigned to an awake or an anesthetized arm. After a baseline period, hemorrhage (7 ml/kg during 20 min) was induced, immediately followed by a 25 ml/kg infusion during 20 min of 0.9% saline. Hemoglobin concentrations, sampled every 5 min for 60 min then every 10 min for an additional 120 min, were used for population kinetic modeling. Covariates, including body weight, sex, and study arm (awake or anesthetized), were tested in the model building. The change in dilution was studied by analyzing area under the curve and maximum plasma dilution. Anesthetized subjects had larger plasma dilution than awake subjects. The analysis showed that females increased area under the curve and maximum plasma dilution by 17% (with 95% CI, 1.08 to 1.38 and 1.07 to 1.39) compared with men, and study arm (anesthetized increased area under the curve by 99% [0.88 to 2.45] and maximum plasma dilution by 35% [0.71 to 1.63]) impacted the plasma dilution whereas a 10-kg increase of body weight resulted in a small change (less than1% [0.93 to 1.20]) in area under the curve and maximum plasma dilution. Mean arterial pressure was lower in subjects while anesthetized (P < 0.001). In awake and anesthetized subjects subjected to controlled hemorrhage, plasma dilution increased with anesthesia, female sex, and lower body weight. Neither study arm nor body weight impact on area under the curve or maximum plasma dilution were statistically significant and therefore no effect can be established.


Serological microarray for a paradoxical diagnostic of Whipple's disease.
Whipple's disease is a systemic chronic infection caused by Tropheryma whipplei. Asymptomatic people may carry T. whipplei in their digestive tract and this can be determined by PCR, making serological diagnosis useful to distinguish between carriers and patients. Putative antigenic proteins were selected by computational analysis of the T. whipplei genome, immunoproteomics studies and from literature. After expression, putative T. whipplei antigens were screened by microimmunofluorescence with sera of immunized rabbit. Selected targets were screened by microarray using sera from patients and carriers. Paradoxically, with 19 tested recombinant proteins and a glycosylated native protein of T. whipplei, a higher immune response was observed with asymptomatic carriers. In contrast, quantification of human IgA exhibited a higher reaction in patients than in carriers against 10 antigens. These results were used to design a diagnostic test with a cut-off value for each antigen. A blind test assay was performed and was able to diagnose 6/8 patients and 11/12 carriers. Among people with positive T. whipplei PCR of the stool, patients differ from carriers by having positive IgA detection and a negative IgG detection. If confirmed, this serological test will distinguish between carriers and patients in people with positive PCR of the stool.


Protective Effect of Alendronate on Lumbar Facet Degeneration in Ovariectomized Rats.
BACKGROUND Facet joint degeneration (FJD) is a potential source of lower back pain, and estrogen deficiency can accelerate FJD. The present study aimed to investigate the effects of alendronate (ALN) on FJD induced by ovariectomy (OVX) in rats. MATERIAL AND METHODS Thirty female Sprague-Dawley rats underwent either bilateral OVX (n=20) or sham surgery (n=10). The OVX rats subsequently received either subcutaneous ALN (70 µg/kg/week) or vehicle for 12 weeks. Subchondral bone mass and microarchitecture were evaluated by micro-computed tomography. Cartilage degradation was evaluated by toluidine blue staining and histological scoring. RESULTS Compared with the Sham group, the OVX group had significantly decreased bone mineral density, bone volume/trabecular volume, and trabecular thickness, significantly increased trabecular separation in subchondral bone, and significantly higher histological score for cartilage degeneration, particularly loss of cartilage thickness. ALN treatment significantly reversed the changes in subchondral bone, preserved cartilage thickness, and reduced the histological score. Immunohistochemical analyses showed significantly decreased expression of ADAMTS-4, MMP-13, and caspase-3 in the OVX+ALN group compared with the OVX group. CONCLUSIONS Treatment with ALN suppressed bone loss, subchondral bone architecture deterioration, and cartilage degeneration in OVX rats, which can be explained by roles of ALN in preservation of subchondral bone mass and microarchitecture, and counteraction of catabolism and chondrocyte apoptosis in cartilage.


Chromosomal breakage and scleroderma: studies in family members.
Chromosome studies were performed on 54 apparently healthy relatives of 29 scleroderma patients and on 40 controls. Increased chromosomal breakage is observed in 86 per cent of brothers and sisters and in 68 per cent of children of patients. If the findings in relatives are compared to those obtained in the 29 scleroderma patients of these families, the percentage of abnormal cells does not differ significantly between the two groups (25.8 per cent and 29.1 per cent respectively), and there is no difference for the frequencies of gaps and open breaks. However acentric fragments, "minutes" and morphologically abnormal chromosomes are significantly increased in patients as compared to their asymptomatic relatives with increased breakage. The distribution of breaks is found to be random. The presence of structural chromosome aberrations in relatives of scleroderma patients may have a special importance with regard to the concept of familial autoimmune disease.


Association between hunting and elevated blood lead levels in the critically endangered African white-backed vulture Gyps africanus.
Lead (Pb) toxicity caused by the ingestion of Pb ammunition fragments in carcasses and offal is a threat to scavenging birds across the globe. African vultures are in critical decline, but research on whether Pb exposure is contributing to declines is lacking. In Africa, recreational hunting represents an important economic activity; however, Pb in leftover hunted carcasses and gut piles represents a dangerous food source for vultures. It is therefore important to establish whether recreational hunting is associated with Pb exposure in African vultures. We explored this issue for the critically endangered white-backed vulture (Gyps africanus) in Botswana by examining their blood Pb levels inside and outside of the hunting season, and inside and outside of private hunting areas. From 566 birds captured and tested, 30.2% birds showed elevated Pb levels (10 to <45 μg/dl) and 2.3% showed subclinical exposure (≥45 μg/dl). Higher blood Pb levels were associated with samples taken inside of the hunting season and from within hunting areas. Additionally, there was a significant interaction between hunting season and areas, with Pb levels declining more steeply between hunting and non-hunting seasons within hunting areas than outside them. Thus, all our results were consistent with the suggestion that elevated Pb levels in this critically endangered African vulture are associated with recreational hunting. Pb is known to be highly toxic to scavenging birds and we recommend that Pb ammunition in Botswana is phased out as soon as possible to help protect this rapidly declining group of birds.


Assessing mechanical function of peristalsis with functional lumen imaging probe panometry: Contraction power and displaced volume.
The distal contractile integral (DCI) quantifies the contractile vigor of primary peristalsis on high-resolution manometry (HRM), whereas no such metric exists for secondary peristalsis on functional lumen imaging probe (FLIP) panometry. This study aimed to evaluate novel FLIP metrics of contraction power and displaced volume in asymptomatic controls and a patient cohort. Thirty-five asymptomatic controls and adult patients (with normal esophagogastric junction outflow/opening and without spasm) who completed HRM and FLIP panometry were included. The patient group also completed timed barium esophagram (TBE). Contraction power (estimate of esophageal work over time) and displaced volume (estimate of contraction-associated fluid flow) were computed from FLIP. HRM was analyzed per Chicago Classification v4.0. In controls, median (5th-95th percentile) contraction power was 27 mW (10-44) and displaced volume was 43 mL (17-66). 95 patients were included: 72% with normal motility on HRM, 17% with ineffective esophageal motility (IEM), and 12% with absent contractility. Among patients, DCI was significantly correlated with both contraction power (rho = 0.499) and displaced volume (rho = 0.342); p values < 0.001. Both contraction power and displaced volume were greater in patients with normal motility versus IEM or absent contractility, complete versus incomplete bolus transit, and normal versus abnormal retention on TBE; p values < 0.02. FLIP panometry metrics of contraction power and displaced volume appeared to effectively quantify peristaltic vigor. These novel metrics may enhance evaluation of esophageal motility with FLIP panometry and provide a reliable surrogate to DCI on HRM.


Effect of starvation and sampling time on plasma alkaline phosphatase activity and calcium homeostasis in the rat.
The effect of starvation and sampling time on plasma alkaline phosphatase activity, total plasma calcium concentration and whole blood ionized calcium concentration was determined in the rat. Starvation caused a significant fall in total and ionized calcium concentrations as well as in alkaline phosphatase activity. These changes were accompanied by a fall in whole blood pH and an increase in the anion gap and a decrease in urinary excretion of calcium. These indices were restored to normal following refeeding. There was no change in serum 25-OH vitamin D concentrations following starvation for 3 days. Alkaline phosphatase activity showed a pattern compatible with the presence of a circadian rhythm when sampling took place between 0800 and 1800 h. Total and ionized calcium concentrations did not show such a rhythm when animals were fed the present diet.


Clostridium difficile infection in hospitalized patients with inflammatory bowel disease: Prevalence, risk factors, and prognosis.
To evaluate the frequency, possible risk factors, and outcome of Clostridium difficile infection (CDI) in inflammatory bowel disease (IBD) patients.There has been an upsurge of CDI in patients with IBD who has been associated with increased morbidity and mortality. Various risk factors have been found to predispose IBD patients to CDI.A retrospective case-control study on IBD patients admitted with exacerbation and tested for CDI at the Tel Aviv Medical Center in 2008 to 2013. Epidemiologic, laboratory, and prognostic data were retrieved from electronic files and compared between patients who tested positive (CDI+) or negative (CDI-) for CDI.CDI was identified in 28 of 311 (7.31%) IBD patients hospitalized with diarrhea. IBD-specific risk factors (univariate analysis) for CDI included: use of systemic steroids therapy (odds ratio [OR] = 3.6, 95% confidence interval [CI] 1.2-10.6) and combinations of ≥2 immunomodulator medications (OR = 2.6, 95% CI 1.1-6.3). Additional risk factors for CDI that are common in the general population were hospitalization in the preceding 2 months (OR = 6.0, 95% CI 2.6-14.1), use of antacids (OR = 3.8, 95% CI 1.7-8.4), and high Charlson comorbidity score (OR = 2.5, 95% CI 1.1-5.7). A multivariate analysis confirmed that only hospitalization within the preceding 2 months and use of antacids were significant risk factors for CDI. The prognosis of CDI+ patients was similar to that of CDI- patients.Hospitalized IBD patients with exacerbation treated with antacids or recently hospitalized are at increased risk for CDI and should be tested and empirically treated until confirmation or exclusion of the infection.


Exciton multiplication from first principles.
Third-generation photovolatics require demanding cost and power conversion efficiency standards, which may be achieved through efficient exciton multiplication. Therefore, generating more than one electron-hole pair from the absorption of a single photon has vast ramifications on solar power conversion technology. Unlike their bulk counterparts, irradiated semiconductor quantum dots exhibit efficient exciton multiplication, due to confinement-enhanced Coulomb interactions and slower nonradiative losses. The exact characterization of the complicated photoexcited processes within quantum-dot photovoltaics is a work in progress. In this Account, we focus on the photophysics of nanocrystals and investigate three constituent processes of exciton multiplication, including photoexcitation, phonon-induced dephasing, and impact ionization. We quantify the role of each process in exciton multiplication through ab initio computation and analysis of many-electron wave functions. The probability of observing a multiple exciton in a photoexcited state is proportional to the magnitude of electron correlation, where correlated electrons can be simultaneously promoted across the band gap. Energies of multiple excitons are determined directly from the excited state wave functions, defining the threshold for multiple exciton generation. This threshold is strongly perturbed in the presence of surface defects, dopants, and ionization. Within a few femtoseconds following photoexcitation, the quantum state loses coherence through interactions with the vibrating atomic lattice. The phase relationship between single excitons and multiple excitons dissipates first, followed by multiple exciton fission. Single excitons are coupled to multiple excitons through Coulomb and electron-phonon interactions, and as a consequence, single excitons convert to multiple excitons and vice versa. Here, exciton multiplication depends on the initial energy and coupling magnitude and competes with electron-phonon energy relaxation. Multiple excitons are generated through impact ionization within picoseconds. The basis of exciton multiplication in quantum dots is the collective result of photoexcitation, dephasing, and nonadiabatic evolution. Each process is characterized by a distinct time-scale, and the overall multiple exciton generation dynamics is complete by about 10 ps. Without relying on semiempirical parameters, we computed quantum mechanical probabilities of multiple excitons for small model systems. Because exciton correlations and coherences are microscopic, quantum properties, results for small model systems can be extrapolated to larger, realistic quantum dots.


Significance of tumor necrosis factor alpha in patients with long-standing type-I diabetes mellitus.
Fifteen out of the forty-five patients with long-standing diabetes mellitus type I were characterised by the presence of TNFalpha in the sera when examined on several occasions over a period of five years. TNFalpha-producing patients had a better control of the disease and a smaller percentage of them suffered from diabetic complications as compared with those not producing this cytokine. TNFalpha-producing patients had lower levels of the proinflammatory markers - IL6 and CRP, and higher concentrations of ACTH and cortisol than those not producing this cytokine. We suggest that TNFalpha released systemically in diabetic patients stimulates the hypophysis-adrenal axis, and in that way indirectly ameliorates autoimmune response occurring during the advanced phase of the disease.


Prevalence of CCR5-Δ32 and CCR2-V64I polymorphisms in a mixed population from northeastern Brazil.
Chemokines are low-molecular weight proteins that play a key role in inflammatory processes. Genomic variations in chemokine receptors are associated with the susceptibility to various diseases. Polymorphisms in chemokine receptor type 5 (CCR5)-Δ32 and CCR2-V64I are related to human immunodeficiency virus infection resistance, which has led to genetic association studies for several other diseases. Given the heterogeneous distribution of these polymorphisms in different global populations and within Brazilian populations, we analyzed the prevalence of CCR5-Δ32 and CCR2-V64I polymorphisms in a mixed population from northeastern Brazil. The study included 223 individuals from the general population of the city of Parnaíba, Piauí, who had a mean age of 73 years. Of these individuals, 37.2% were men and 62.8% were women. Polymorphisms were analyzed using DNA extracted from peripheral blood leukocytes by using polymerase chain reaction alone (CCR5-Δ32) or accompanied by restriction endonuclease digestion (CCR2-V64I). In both cases, the genotypes were determined using 8% polyacrylamide gel electrophoresis and silver nitrate staining. The population conformed to Hardy-Weinberg equilibrium for both the loci studied. No individuals were homozygous for allele-Δ32, which was present in 1.8% of the population, whereas allele-64I was present in 13.9% of the participants studied; 74.9% were homozygous for the wild-type allele, while 22.4 and 2.7% were heterozygous and homozygous for the mutant allele, respectively. Additional studies are needed to investigate the relationship between these polymorphisms and disease etiopathogenesis in reference populations.


Genetics of gynaecological disorders.
From genomic imbalances associated with developmental abnormalities of the female genital tract to the molecular mechanisms underpinning endometriosis and uterine leiomyomatosis, new technologies have allowed the exploration of the genetic contribution and mapping the molecular pathways underpinning common and rare gynaecological conditions. While some of these conditions have historically been considered sporadic, recent research has demonstrated their potentially heritable nature linked to single genes or copy number variants. The phenotypic variability including non-penetrance indicates their multifactorial, complex aetiology encompassing genetic, epigenetic and environmental influences. Although genetic tests are not routinely conducted in gynaecological practice, there is an increasing body of evidence suggesting that, in appropriate cases, molecular investigations such as array CGH analysis may be an important part of the diagnostic algorithm. The subtlety of clinical features, especially in the context of syndromic diagnoses, requires the practitioner to become familiar with those conditions and the approach to diagnostic investigations. This chapter combines the recent research output related to gynaecological disorders with a clinical genetics approach aiming to highlight the multisystem character of some of these conditions, their implications for management, reproductive risks and options, and the importance of genetic counselling.


Dispersion of Complex Refractive Indices for Intense Vibrational Bands. I. Quantitative Spectra.
This paper reviews the dispersion of complex refractive indices for representative intense vibrational bands of organic liquids in the absolute scale. Conspicuous variations in both real and imaginary refractive indices over the intense bands are precisely determined as a function of infrared wavenumber by attenuated total reflection infrared absorption measurement. The accurate spectral data offer an excellent reference to calibrate the absolute infrared intensities by various quantum chemical calculations, and thus, critical comparison between the present experiment and computation was reported to clarify the general accuracy of the quantum chemical calculations. The precise data of the dispersion will be utilized to clarify their impact on the analysis of vibrational spectroscopy of interfaces in the subsequent paper.


Crystalline phases of polydisperse spheres.
We use specialized Monte Carlo simulation methods and moment free energy calculations to provide conclusive evidence that dense polydisperse spheres at equilibrium demix into coexisting fcc phases, with more phases appearing as the spread of diameters increases. We manage to track up to four coexisting phases. Each of these is fractionated: it contains a narrower distribution of particle sizes than is present in the system overall. We also demonstrate that, surprisingly, demixing transitions can be nearly continuous, accompanied by fluctuations in local particle size correlated over many lattice spacings.


Ventilatory failure during loaded breathing: the role of central neural drive.
Minute ventilation (VE), arterial blood gases, diaphragmatic electromyogram (EMG) activity, centroid frequency (Fc) and peak inspiratory airway pressures (Paw) were measured in five unanesthetized tracheostomized infant monkeys during various intensities of inspiratory resistive loaded breathing (IRL) until either 1) ventilatory failure occurred (failed trial) or 2) normocapnia was sustained for 1 h (successful trial). During successful trials VE and arterial PCO2 (PaCO2) were sustained at base-line levels, and an increase in peak integrated diaphragmatic EMG activity and peak inspiratory Paw occurred. In contrast, during ventilatory failure runs, VE decreased and PaCO2 rose compared with their respective base-line values. The fall in VE occurred secondary to a significant decline in breathing frequency. Tidal volume was sustained at base-line levels during all trials (both successful and failed groups). Inspiratory Paw's and peak moving time average EMG were sustained at elevated levels during ventilatory failure runs, suggesting that the respiratory muscles did not fail as pressure generators. Furthermore, the EMG Fc did not change from base line during either successful or failed trials. These data suggest that peripheral muscle fatigue did not occur, although in the absence of a more direct test of muscle performance, i.e., a force-frequency curve, we cannot rule out the possibility that a component of peripheral failure contributed to our results. Ventilatory failure during severe IRL in the infant monkey was most clearly associated with an alteration in the respiratory center timing mechanism, i.e., such failure was a function of a decline in respiratory frequency.


Constructing van der Waals Heterogeneous Photocatalysts Based on Atomically Thin Carbon Nitride Sheets and Graphdiyne for Highly Efficient Photocatalytic Conversion of CO2 into CO.
Atomically thin two-dimensional (2D) carbon nitride sheets (CNs) are attracting attention in the field of photocatalytic CO2 reduction. Because of the rapid recombination of photogenerated electron-hole pairs and limited more active sites, the photocatalytic efficiency of CNs cannot meet the actual requirements. Here, atomically thin 2D/2D van der Waals heterostructures of metal-free graphdiyne (GDY)/CNs are fabricated through a simple electrostatic self-assembly method. Experimental characterizations along with first-principles calculations show that the introduction of GDY in CNs promoted the transport of photogenerated carriers in the melon chain, thus suppressing the recombination of photogenerated electron-hole pairs. Both in situ FTIR measurements and DFT calculation confirm that the introduced GDY served as the CO2 adsorption site and enhanced the CO2 adsorption capacity of the CNs/GDY heterostructure. Thanks to the 2D/2D van der Waals heterojunction, the optimized CNs/GDY enhances significantly the CO generation rate up to 95.8 μmol g-1 that is 19.2-fold higher than that of CNs. This work provides a viable approach for the design of metal-free van der Waals heterostructure-based photocatalysts with high catalytic activity.


Social factors and leprosy in Lamjung, West Central Nepal: implication for disease control.
Such is the ability of leprosy to generate misconceptions and fears, that many patients are reluctant to be identified. Deformity and paralysis which may occur compound the stigma attached to this rare disease of slow insidious onset. Epidemiological studies of leprosy refer only to known disease and often to highly selected groups of the population. Cohorts are therefore incomplete, and variations in prevalence may reflect social attitudes and data reliability. This paper describes the demographic and spatial distribution of leprosy in Lamjung, a district of west central Nepal. Variations in known leprosy prevalence between sexes, ethnic groups and areas are related to social and physical factors. An apparent paradox of low leprosy prevalence in an ethnic group with a high proportion of infectious leprosy is associated with adverse social attitudes and poor survey coverage. Although the data are too limited for an epidemological analysis variations associated with social and physical factors have crucial implications for disease control.


Assessing the spatial distribution of Tuta absoluta (Lepidoptera: Gelechiidae) eggs in open-field tomato cultivation through geostatistical analysis.
The spatial distribution of insects is due to the interaction between individuals and the environment. Knowledge about the within-field pattern of spatial distribution of a pest is critical to planning control tactics, developing efficient sampling plans, and predicting pest damage. The leaf miner Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is the main pest of tomato crops in several regions of the world. Despite the importance of this pest, the pattern of spatial distribution of T. absoluta on open-field tomato cultivation remains unknown. Therefore, this study aimed to characterize the spatial distribution of T. absoluta in 22 commercial open-field tomato cultivations with plants at the three phenological development stages by using geostatistical analysis. Geostatistical analysis revealed that there was strong evidence for spatially dependent (aggregated) T. absoluta eggs in 19 of the 22 sample tomato cultivations. The maps that were obtained demonstrated the aggregated structure of egg densities at the edges of the crops. Further, T. absoluta was found to accomplish egg dispersal along the rows more frequently than it does between rows. Our results indicate that the greatest egg densities of T. absoluta occur at the edges of tomato crops. These results are discussed in relation to the behavior of T. absoluta distribution within fields and in terms of their implications for improved sampling guidelines and precision targeting control methods that are essential for effective pest monitoring and management. © 2017 Society of Chemical Industry.


Lipid emulsion improves survival in animal models of local anesthetic toxicity: a meta-analysis.
The Lipid Emulsion Therapy workgroup, organized by the American Academy of Clinical Toxicology, recently conducted a systematic review, which subjectively evaluated lipid emulsion as a treatment for local anesthetic toxicity. We re-extracted data and conducted a meta-analysis of survival in animal models. We extracted survival data from 26 publications and conducted a random-effect meta-analysis based on odds ratio weighted by inverse variance. We assessed the benefit of lipid emulsion as an independent variable in resuscitative models (16 studies). We measured Cochran's Q for heterogeneity and I2 to determine variance contributed by heterogeneity. Finally, we conducted a funnel plot analysis and Egger's test to assess for publication bias in studies. Lipid emulsion reduced the odds of death in resuscitative models (OR =0.24; 95%CI: 0.1-0.56, p = .0012). Heterogeneity analysis indicated a homogenous distribution. Funnel plot analysis did not indicate publication bias in experimental models. Meta-analysis of animal data supports the use of lipid emulsion (in combination with other resuscitative measures) for the treatment of local anesthetic toxicity, specifically from bupivacaine. Our conclusion differed from the original review. Analysis of outliers reinforced the need for good life support measures (securement of airway and chest compressions) along with prompt treatment with lipid.


Ocular Findings in the 16p11.2 Microdeletion Syndrome: A Case Report and Literature Review.
The recurrent 16p11.2 microdeletion is characterized by developmental delays and a wide spectrum of congenital anomalies. It has been well reported that individuals with this ∼593-kb interstitial deletion have an increased susceptibility toward the autism spectrum disorder (ASD). Abnormalities of the eye and ocular adnexa are also commonly associated findings seen in individuals with the 16p11.2 microdeletion syndrome, although these ophthalmic manifestations have not been well characterized. We conducted an extensive literature review to highlight the eye features in patients with the 16p11.2 microdeletion syndrome and describe a 5-year-old boy with the syndrome. The boy initially presented with intellectual disability, speech delay, and defiant behavior; diagnoses of attention deficit hyperactivity disorder (ADHD) and oppositional defiant disorder (ODD) were established. He had a Chiari malformation type 1. His ophthalmic features included strabismus, hyperopia, and ptosis, and a posterior embryotoxon was present bilaterally. From a systematic review of prior reported cases, the most common eye and ocular adnexa findings observed were downslanting palpebral fissures, deep-set eyes, ptosis, and hypertelorism.


Association between subcellular proliferating membranes and mitochondria in dog thyroid follicular cells induced by synthetic double-stranded RNA administration.
The administration of polyriboinosinic-polyribocytidylic acid [poly(rI:rC)], an interferon inducer and antitumour agent, induced proliferating intracellular membranes as well as a characteristic association between those membranes and mitochondria. The membrane-mitochondrion association was most often observed in small mitochondria lying close to the basal cell membrane. Although the meaning of this differential response is not known, these findings might be explained by an actual biogenesis of the mitochondria from subcellular membranes induced by poly (rI:rC), either directly or secondary to its toxic effects.


[Characterization of a T-cell subpopulation with a new monoclonal antibody BL-TSub/2].
The mab BL-TSub/2 binds to a majority (80%) of thymocytes. The antigen isolated from these cells has an apparent molecular weight of about 170 kDa. The mab BL-TSub/2 recognized antigen is expressed on approximately 50% of mature T cells in peripheral blood. The mab BL-TSub/2 does not bind to monocytes, granulocytes, platelets and CD16+NK cells. The antigen is possibly present on a part of peripheral B cells in very low density. Double labelling experiments showed that about 30% of CD4+ and 60% of CD8+ cells react with the mab BL-TSub/2. This BL-TSub/2+ subset is not identical with T-cell subpopulations till now defined. Within the CD4+ subset the majority of BL-TSub/2+ cells is included in the CD45RO+ subset while only a small proportion is CD45RA+. However, the CD8+ BL-TSub/2+ cells are mostly CD45RA+ with only few cells belonging to the CD45RO+ subset.


Synthetic Oligodeoxynucleotides Containing Multiple Telemeric TTAGGG Motifs Suppress Inflammasome Activity in Macrophages Subjected to Oxygen and Glucose Deprivation and Reduce Ischemic Brain Injury in Stroke-Prone Spontaneously Hypertensive Rats.
The immune system plays a fundamental role in both the development and pathobiology of stroke. Inflammasomes are multiprotein complexes that have come to be recognized as critical players in the inflammation that ultimately contributes to stroke severity. Inflammasomes recognize microbial and host-derived danger signals and activate caspase-1, which in turn controls the production of the pro-inflammatory cytokine IL-1β. We have shown that A151, a synthetic oligodeoxynucleotide containing multiple telemeric TTAGGG motifs, reduces IL-1β production by activated bone marrow derived macrophages that have been subjected to oxygen-glucose deprivation and LPS stimulation. Further, we demonstrate that A151 reduces the maturation of caspase-1 and IL-1β, the levels of both the iNOS and NLRP3 proteins, and the depolarization of mitochondrial membrane potential within such cells. In addition, we have demonstrated that A151 reduces ischemic brain damage and NLRP3 mRNA levels in SHR-SP rats that have undergone permanent middle cerebral artery occlusion. These findings clearly suggest that the modulation of inflammasome activity via A151 may contribute to a reduction in pro-inflammatory cytokine production by macrophages subjected to conditions that model brain ischemia and modulate ischemic brain damage in an animal model of stroke. Therefore, modulation of ischemic pathobiology by A151 may have a role in the development of novel stroke prevention and therapeutic strategies.


Editorial -- Reducing premature mortality among young and middle-aged adults.
The death of an individual in early and middle adulthood is an untimely event whose tragic effects are experienced by the parents, siblings, partners, children and friends of the deceased individual. Preventing premature death is a foremost goal of health care and public health programs, and of society at large.


Induction of indoleamine 2,3-dioxygenase in vascular smooth muscle cells by interferon-gamma contributes to medial immunoprivilege.
Atherosclerosis and graft arteriosclerosis are characterized by leukocytic infiltration of the vessel wall that spares the media. The mechanism(s) for medial immunoprivilege is unknown. In a chimeric humanized mouse model of allograft rejection, medial immunoprivilege was associated with expression of IDO by vascular smooth muscle cells (VSMCs) of rejecting human coronary artery grafts. Inhibition of IDO by 1-methyl-tryptophan (1-MT) increased medial infiltration by allogeneic T cells and increased VSMC loss. IFN-gamma-induced IDO expression and activity in cultured human VSMCs was considerably greater than in endothelial cells (ECs) or T cells. IFN-gamma-treated VSMCs, but not untreated VSMCs nor ECs with or without IFN-gamma pretreatment, inhibited memory Th cell alloresponses across a semipermeable membrane in vitro. This effect was reversed by 1-MT treatment or tryptophan supplementation and replicated by the absence of tryptophan, but not by addition of tryptophan metabolites. However, IFN-gamma-treated VSMCs did not activate allogeneic memory Th cells, even after addition of 1-MT or tryptophan. Our work extends the concept of medial immunoprivilege to include immune regulation, establishes the compartmentalization of immune responses within the vessel wall due to distinct microenvironments, and demonstrates a duality of stimulatory EC signals versus inhibitory VSMC signals to artery-infiltrating T cells that may contribute to the chronicity of arteriosclerotic diseases.


Studies on the cytotoxicity, mutagenicity and chromosomal aberration-inducing activity of aclacinomycin A in cultured mammalian cells.
The cytotoxicity mutagenicity and chromosomal aberration-inducing activity of aclacinomycin A were studied using cultured mammalian cells. Aclacinomycin A was found to be a potent inhibitor of RNA synthesis in HeLa S3 cells. The survival curve in the presence of aclacinomycin A exhibited a concentration- and time-dependent cytocidal effect, and showed a sigmoidal shoulder. A cytofluorometric study suggested that aclacinomycin A caused a temporary blockage of cell cycle progression. In mutation assay using V79 cells, aclacinomycin A was shown to induce 8-azaguanine-resistant mutation, being several times more mutagenic than adriamycin. The induction of chromosomal aberrations by aclacinomycin A was also demonstrated. When cells from various sources were treated with aclacinomycin A, they were affected at a similar concentration with the exception of human normal fibroblasts, which were a little less sensitive than the others.


Olfactomedin-like 3: possible functions in embryonic development and tumorigenesis.
Modern medical research has proven that human diseases are directly or indirectly related to genes. At the same time, genetic research has also brought updates to diagnostic techniques. Olfactomedin-like 3 (OLFML3) gene is a novel and clinically valuable gene. In order to better understand the role of OLFML3 in human diseases, we discuss and analyze the characteristics, function, and regulation mechanism of the OLFML3 gene in this review. A comprehensive search in PubMed and ScienceDirect database for English up to March 2019, with the keywords of "Olfactomedin-like 3," "Olfactomedin," "extracellular matrix," "Transforming Growth Factor β1," "anoikis-resistance," and "microRNA-155." Careful review of all relevant literature, the references of the retrieved articles were also screened to search for potentially relevant papers. OLFML3 is a secreted glycoprotein with 406 amino acid residues, belonging to the Olfactomedin (OLF) family. Due to the particularity of its structure and differential expression, OLFML3 has unique biological functions that could be distinct from other members in the OLF family. The currently known functions include embryonic development function and tumorigenesis. The regulation mechanism is still under investigation. It is directly related to many human diseases. OLFML3 is a multifunctional glycoprotein that is closely involved in embryonic development, tumor invasion, and metastasis. Unfortunately, current research on this important molecule is still very limited. Further investigations on the possible mechanism of OLFML3 biological functions and modulation will help us develop better diagnostics and treatments.


Prevalence of Function-Dependent Temporomandibular Joint and Masticatory Muscle Pain, and Predictors of Temporomandibular Disorders among Patients with Lyme Disease.
The aim was to determine the occurrence of temporomandibular disorders (TMDs) in patients with Lyme disease (LD), and to estimate the contribution of factors that may identify TMD among LD patients. In seventy-six (N = 76) adult patients with LD (mean age 57.6 ± 14.6 years) and 54 healthy non-Lyme volunteers with a mean age of 56.4 ± 13.5 years, possible function (i.e., non-pain) diagnoses were established using the Research Diagnostic Criteria of Temporomandibular Disorders (RDC/TMD). Pain diagnoses were established by means of the function-dependent dynamic and static tests. The two groups did not significantly differ in the frequency of disc displacements diagnoses and function-dependent pain diagnoses. LD showed a significantly higher frequency (p < 0.001) of osteoarthrosis than the control group. For the prediction of pain diagnoses in LD patients, the single regression analyses pointed out an association with age, sleep bruxism (SB), and awake bruxism (AB). Two predictors (i.e., SB (p = 0.002) and AB (p = 0.017)) were statistically significant in the final multiple variable model. The frequency of TMD in patients with LD based on function-dependent tests was not significantly different from that in the control group. This investigation suggests that the contribution of bruxism to the differentiation between patients with Lyme and TMD is high.


Development of a molecular assay specific for the Leishmania donovani complex that discriminates L. donovani/Leishmania infantum zymodemes: a useful tool for typing MON-1.
We have developed a simple, rapid, sensitive, and cost-effective typing method, based on the amplicon size of the K26 gene, capable of species/strain discrimination of Leishmania donovani complex strains causing visceral leishmaniasis (VL). It was evaluated on 112 strains and compared with multilocus enzyme electrophoresis (MLEE) typing. The K26 polymerase chain reaction (PCR) applied on 26 representative L. donovani complex strains gave 14 different amplicon sizes. The assay was specific to the L. donovani complex and discriminated L. infantum from L. donovani strains. MON-1 strains were also easily distinguished from other non-MON-1. Surprisingly, 29.3% of the Greek strains included in this study were MLEE typed as MON-98 and gave exclusively a 940-bp amplicon. The majority of Greek MON-1 strains gave also the 940-bp amplicon, whereas a 626-bp amplicon was consistently obtained with other European MON-1 strains. K26 PCR-restriction fragment length polymorphism, based on MON-1 K26 sequence polymorphism, gave 2 MON-1 subgroups. Application of the method may contribute to efficiently monitor VL.


Iliac Crest Avulsion Fracture in a Young Sprinter.
Avulsion fracture of the iliac crest is an uncommon pathology. It usually occurs in teenagers during sport activities, more common in boys. We report a case of 16-year-old male competitive sprinter, who had an avulsion of a part of the iliac crest and the anterior-superior iliac spine during a competition. The traumatism occurred during the period of acceleration phase out of the blocks which corresponds to the maximum traction phase on the tendons. Then a total loss of function of the lower limb appears forcing him to stop the run. X-ray and CT scan confirmed the rare diagnosis of avulsion of the quasitotality of the iliac crest apophysis, corresponding to Salter 2 fracture. We performed an open reduction and internal fixation with two screws, allowing a return to sport after 3 months and his personal best record in the 100 meters at the 6th postoperative month.


[Myocardial remodeling in patients with chronic heart failure and implanted cardiac contractility modulators].
To study the effect of cardiac contractility modulation on reverse remodeling and myocardial function in patients with chronic heart failure (HF) according to echocardiography (EchoCG). In a group of 40 patients with a combination of chronic HF and atrial fibrillation (AF), the dynamics of standard EchoCG parameters and the effectiveness of myocardial work of the left ventricle (LV) against the background of 12-month therapy of cardiac contractility modulation (CCM) were analyzed. The results obtained indicate a statistically significant positive effect of CCM on LV remodeling parameters and the effectiveness of myocardial work according to EchoCG. Transthoracic echocardiography is the main imaging method and provides great opportunities for evaluating the effectiveness of HF treatment, including non-drug methods such as CCM. Evaluation of LV myocardial function in patients with HF and implanted CCM devices is a promising scientific and practical research method.


15-deoxy-(Delta12,14)-prostaglandin J2 (15d-PGJ2) induces cell death through caspase-independent mechanism in A172 human glioma cells.
15-Deoxy-(Delta12,14)-prostaglandin J(2) (15d-PGJ(2)) is a naturally occurring cyclopentenone metabolite of prostaglandin D(2) (PGD(2)) and is known as a specific potent ligand for the peroxisome proliferators activator receptor-gamma (PPARgamma). 15d-PGJ(2) inhibits cell growth and induces apoptosis in a number of different cancer cells. However, the underlying mechanism by which 15d-PGJ(2) induces cell death remains to be defined. The present study was undertaken to determine the effect of 15d-PGJ(2) on cell death in A172 human glioma cells. 15d-PGJ(2) caused reactive oxygen species (ROS) generation. 15d-PGJ(2)-induced ROS production and cell death were prevented by the antioxidant N-acetylcysteine. Activation of mitogen-activated protein kinases (MAPK) was not observed in cells treated with 15d-PGJ(2 )and inhibitors of MAPK subfamilies also were not effective in preventing 15d-PGJ(2)-induced cell death. 15d-PGJ(2) treatment caused mitochondrial dysfunction, as evidenced by depolarization of mitochondrial membrane potential. 15d-PGJ(2) induced caspase activation at 24 h of treatment, but the 15d-PGJ(2)-induced cell death was not prevented by caspase inhibitors. The antiapoptotic protein XIAP levels and release of apoptosis inducing factor (AIF) into the cytosol were not altered by 15d-PGJ(2) treatment. Taken together, these findings indicate that 15d-PGJ(2) triggers cell death through a caspase-independent mechanism and ROS production and disruption of mitochondrial membrane potential play an important role in the 15d-PGJ(2)-induced cell death in A172 human glioma cells.


Development of microwave imaging reflectometry on the HL-2A tokamak.
A microwave imaging reflectometry system has been developed to visualize the density fluctuations on the HL-2A tokamak. This system is characterized by a quasi-optical system, a four frequency microwave transmitter, and a microwave quadrature receiver system with a 3D adjustable U-shaped horn antenna array, that generate 8 (poloidal) × 4 (radial) × 2 (toroidal) = 64 channel images of density fluctuations. Simulations and laboratory tests of the optical system have been conducted. The test results are in good agreement with the simulations.


Goals of care and end-of-life communication needs of persons with chronic respiratory disease.
To highlight recent advances in effective communication among persons with chronic respiratory diseases. The authors focus on communication science related to goals of care (GOC) discussions, medical devices, and life-sustaining invasive treatments. The authors discuss important considerations when working with individuals with low literacy and rurality. Communication handoffs between respiratory clinicians and/or palliative care to hospice clinicians are summarized to ensure effective person-centered and caregiver-centered care. Studies suggest the following communication approaches: (1) clarify differences between palliative and end of life; (2) conduct conversations early and gradual throughout the illness trajectory; (3) distinguish types of GOC discussions as they relate to treatment preferences; (4) for patients from rural communities, include family members and spiritual leaders; (5) assess literacy and employ supportive strategies; (6) apply time-limited-trial framework for life-sustaining treatment (LST) decisions; and (7) standardize processes for communication handoffs to hospice clinicians to improve communication fidelity. Effective communication tools for clinicians to engage in GOC discussions for persons with chronic respiratory diseases are grounded in a patient-centered framework. A trained clinician should lead these conversations and include interdisciplinary team members throughout the disease trajectory including at the end of life. These approaches may enable patients to express their values and care preferences as they evolve over time.


Speculations on disease states induced by excitatory amino acids.
There is much current interest in excitatory amino acids and their receptors because of their postulated involvement in several disorders of the nervous system. They function as neurotransmitters, but can act as neurotoxins in some situations. They have been implicated in the pathogenesis of cerebral hypoxic/ischemic and hypoglycemic damage, in epilepsy, in some degenerative diseases, and in some forms of neurotoxin-induced cerebral dysfunction. These diseases may reflect abnormality in a system which has evolved to provide synaptic plasticity essential for learning and memory. The purpose of this paper is to explore the ramifications of such a hypothesis.


Long-Term Impacts of COVID-19 Pneumonia on Quality of Life: A Single Institutional Pilot Study.
Many studies have demonstrated poor quality of life (QoL) at 3, 6, 12, and 24 months after coronavirus disease 2019 (COVID-19). However, these studies were limited due to cross-sectional design, a longer gap between visits, and lack of controls for comparison. Therefore, the aim of our prospective study was to assess the impact of COVID-19 pneumonia on QoL in both physical and mental health. A prospective study was conducted on adult patients with COVID-19 pneumonia. We used the 36-Item Short Form Health Survey (SF-36) and Euro Quality of Life-5 Dimensions-5 Levels (EQ-5D-5L), EQ visual analogue scale (EQ-VAS), and Hospital Anxiety and Depression Scale to collect data at months, 1, 3, 6, 9, and 12. Thirty-eight patients with COVID-19 pneumonia and twenty-five healthy subjects were completely followed up on all visits. All domains of SF-36, except bodily pain and EQ-5D-5L of the patients, were lower than controls. There was an improvement of EQ-VAS and SF-36 including physical functioning, social functioning, and role limitation (physical problems) domains throughout study period in the COVID-19 pneumonia group. Adult patients who recovered from COVID-19 pneumonia had lower QoL which improved over the one-year follow-up period.


Prevalence of hepatitis B and hepatitis C virus coinfection in chronic liver disease.
Coinfection with HBV and HCV may lead to serious consequences. The present study was done to find out the prevalence of coinfection in patients with chronic liver disease. From patients with hepatitis and chronic liver disease 1673 samples were received and analysed for HBsAg by ELISA. 1342 samples were analysed for anti HCV by third generation ELISA. 493 samples positive for HBsAg were also analysed for Anti HCV to see the prevalence of coinfection. 15(3.0%) were found positive for both HBsAg and anti HCV. Out of 15 patients with coinfection 4 (26.6%) had HCC. Prevalence of HCC in patients with coinfection was higher than either infection alone i.e. HBV-9.1% and HCV-16.5%.


Quantitative multiple reaction monitoring of peptide abundance introduced via a capillary zone electrophoresis-electrospray interface.
We demonstrate the use of capillary zone electrophoresis with an electrokinetic sheath-flow electrospray interface coupled to a triple-quadrupole mass spectrometer for the accurate and precise quantification of Leu-enkephalin in a complex mixture using multiple-reaction monitoring (MRM). Assay time is <6 min, with no re-equilibration required between runs. A standard curve of Leu-enkephalin was performed in the presence of a background tryptic digest of bovine albumin. We demonstrate reasonably reproducible peak heights (21% relative standard deviation), retention times (better than 1% relative standard deviation), and robust electrospray quality. Our limit of detection (3σ) was 60 pM, which corresponds to the injection of 335 zmol of peptide. This is a 10-20-fold improvement in mass sensitivity than we have obtained by nano HPLC/MRM and substantially better than reported for LC/MS/MS. Further quantification was performed in the presence of stable-isotope-labeled versions of the peptides; under these conditions, linearity was observed across nearly 4 orders of magnitude. The concentration detection limit was 240 pM for the stable-isotope-labeled quantification.


Advances in the application of high-throughput sequencing in invertebrate virology.
Over the last decade, advances in high-throughput sequencing technologies have revolutionised biological research, making it possible for DNA/RNA sequencing of any organism of interest to be undertaken. Sequencing approaches are now routinely used in the detection and characterisation of (novel) viruses, investigation of host-pathogen interactions, and effective development of disease treatment strategies. For the sequencing and identification of viruses of interest, metagenomics approaches using infected host tissue are frequently used, as it is not always possible to culture and isolate these pathogens. High-throughput sequencing can also be used to investigate host-pathogen interactions by investigating (temporal) transcriptomic responses of both the host and virus, potentially leading to the discovery of novel opportunities for treatment and drug targets. In addition, viruses in environmental samples (e.g. water or soil samples) can be identified using eDNA/metagenomics approaches. The promise that recent developments in sequencing brings to the field of invertebrate virology are not devoid of technical challenges, including the need for better laboratory and bioinformatics strategies to sequence and assemble virus genomes within complex tissue or environmental samples, and the difficulties associated with the annotation of the large number of novel viruses being discovered.


Emerging Roles of SIRT3 in Cardiac Metabolism.
The heart is a highly metabolically active organ that predominantly utilizes fatty acids as an energy substrate. The heart also derives some part of its energy by oxidation of other substrates, including glucose, lactose, amino acids and ketones. The critical feature of cardiac pathology is metabolic remodeling and loss of metabolic flexibility. Sirtuin 3 (SIRT3) is one of the seven mammalian sirtuins (SIRT1 to SIRT7), with NAD+ dependent deacetylase activity. SIRT3 is expressed in high levels in healthy hearts but downregulated in the aged or diseased hearts. Experimental evidence shows that increasing SIRT3 levels or activity can ameliorate several cardiac pathologies. The primary deacetylation targets of SIRT3 are mitochondrial proteins, most of which are involved in energy metabolism. Thus, SIRT3 improves cardiac health by modulating cardiac energetics. In this review, we discuss the essential role of SIRT3 in regulating cardiac metabolism in the context of physiology and pathology. Specifically, we summarize the recent advancements that emphasize the critical role of SIRT3 as a master regulator of cardiac metabolism. We also present a comprehensive view of all known activators of SIRT3, and elaborate on their therapeutic potential to ameliorate energetic abnormalities in various cardiac pathologies.


A Review of Hypnotherapy for Overactive Bladder.
This review examines the effectiveness of hypnotherapy for the relief of overactive bladder (OAB) symptoms. Ten studies examining outcomes of hypnotherapy for OAB were located from searches of electronic databases. Most reports were case studies or observational, but there were two randomized, controlled trials. Hypnotherapeutic treatment regimens were idiosyncratic and tailored to individual patients. All studies suggested benefits from hypnotherapy as an adjunct treatment for OAB, especially in terms of subjective reports of symptoms and increasing self-efficacy. These benefits suggest hypnotherapy increases patients' abilities to engage in relaxation, reduces condition-associated anxiety, and improves patients' perceptions of their symptom-coping abilities. Although strong objective evidence of improvement in OAB symptoms is lacking, these subjective improvements, combined with increasing use and acceptance of hypnotherapy in obstetric and gynecological settings, suggest the utility of hypnotherapy as a psychological adjunctive procedure in the treatment of OAB.


Embryo donation or embryo adoption? Conceptual and normative issues.
A central question in the ethical debate on the practice of relinquishing in vitro fertilization surplus embryos for family building is whether we ought to think of it more in terms of donating these embryos or in terms of having them adopted. Deciding between these two alternatives is more than a matter of mere terminology. It has an impact on normative questions, e.g., on the question of what criteria for parent selection ought to be applied to the recipients of the embryos, and on the moral evaluation of the act of 'donating' the embryo or 'having it adopted'. In this article, I defend the view that we should conceptualize the relinquishment of spare embryos according to the adoption model, not as a donation. Section 2 sketches the outline of the argument by making clear how we may ground a defense of the adoption model in a theory of parental responsibility without implicitly elevating the moral status of the embryo. Section 3 contains a preliminary defense of the adoption model that draws on geneticism as what seems to me the most persuasive theory of parental responsibility. In section 4, I examine three objections to geneticism and either rebut them or, insofar as they are justified, try to accommodate them into my view. In section 5, I point out some features that distinguish embryo adoption from the adoption of (born) children. I contend, however, that these differences are compatible with the adoption model. Section 6 is concerned with the normative ramifications of this view.


Chimerism and use of mesenchymal stem cells in umbilical cord blood transplantation.
We performed a retrospective single-center analysis of 50 umbilical cord blood transplantations (UCBTs), focusing on chimerism development. Complete donor chimerism (DC) was associated with acute graft-vs.-host disease (aGVHD) grades II-IV for the CD3 (+) cell lineage (p = 0.01) and, in multivariate analysis, with total body irradiation (TBI) for all lineages (p < 0.01). Overall survival (OS) was negatively associated with patient age, (p < 0.001); aGVHD grades III-IV, (p < 0.001); and treatment with mesenchymal stem cells (MSCs) (p = 0.027). In conclusion, though multiple factors may have contributed, the association of TBI and DC might be worthy of consideration in the treatment of patients with malignant disease in the UCBT setting. The negative influence of MSCs on OS may be a reason for more careful usage of this treatment modality in combination with UCBT.


Magnesium: the fifth but forgotten electrolyte.
Magnesium (Mg) is the second most abundant intracellular cation and is a cofactor in more than 300 enzymatic reactions involving energy metabolism and protein and nucleic acid synthesis. Ionized Mg is the physiologically active form of the element. Protein-bound and chelated Mg buffer the ionized pool. Approximately half the total Mg in the body is present intracellularly in soft tissue, and the other half is present in bone. Less than 1% of the total body Mg is present in blood. However, the majority of our clinical laboratory information comes from the determination of total Mg in serum. Currently, the clinical laboratory evaluation of Mg status is limited primarily to the total serum Mg concentration and a 24-hour urinary excretion. Instrumentation to determine ionized Mg in serum (ion-selective electrode) and in soft tissue (nuclear magnetic resonance spectroscopy) should be available in the near future. Magnesium may be a factor in the treatment of acute myocardial infarction and the rate of atherosclerosis. Chronic changes of Mg status, that may be latent, are poorly understood and require a better knowledge of ionized Mg metabolism.


Nanoparticulate coatings with efficient up-conversion properties.
Nanoparticulate films with high up-conversion emission (UC) properties were prepared by spray-deposition of nanometer-sized YVO4:Yb,Er particles. The optical properties of YVO4:Yb,Er were optimized upon annealing before the film deposition in order to get the highest possible UC signal in the considered type of system. Thanks to a simple model and some time-resolved spectroscopic investigations, the contribution of the scattering to the UC signal could be separated from the intrinsic properties (crystallinity, surface defects) of the material. The films obtained by this technique present the advantages of having both high UC and good transparency.