Comparative studies with three-week-old chickens, turkeys, ducks, and quails on the response in phosphorus utilization to a supplementation of monobasic calcium phosphate.
We studied whether the availability of P is different among poultry species. A basal diet was mixed mainly based on corn, potato protein, and dried egg white. It was calculated according to the recommendations for young turkeys with the exception of P and Ca concentrations, which were deficient. Monobasic calcium phosphate (MCP) was added in graded levels, and analyzed P concentrations in the 7 diets were (in g/kg of dry matter) 2.9, 3.7, 4.5, 5.3, 6.2, 7.2, and 7.7. Four experiments were conducted with 3-wk-old broiler chickens, turkeys, ducks, and quails. Birds were kept in balance crates, and 8 individuals were allocated to each treatment diet. Birds were fed the treatment diets for 10 d, and excreta were quantitatively collected during the last 5 d. P utilization was calculated as the proportion of P intake that was accreted by the birds. The P accretion response of birds to incremental MCP intake was described with sigmoidal functions, and the marginal efficiency of P utilization ((delta)y/(delta)x) was calculated. Utilization of P from the unsupplemented basal diet was 58% in broilers, 55% in quails, 46% in ducks, and 39% in turkeys. Supplementation of MCP significantly increased P accretion in all species. Ninety-five percent of the estimated ymax in P accretion was achieved with 8.4, 7.3, and 4.8 g P/kg of dietary DM in broilers, ducks, and quails. No plateau in P accretion was achieved in turkeys. These differences correlate well with the differences in the feed/gain ratio, which was 1.3 in turkeys, 1.7 in broilers, 1.9 in ducks, and 3.5 in quails. The maximum in marginal efficiency of supplemented P was 96% in ducks, 81% in turkeys, 74% in broilers, and 77% in quails. These maxima were achieved at different levels of MCP supplementation. We concluded that differences in P availability exist between poultry species for plant and mineral P sources. Quails can be used as model species for broilers in P availability studies, but dietary P levels need special adjustment.


The Effects of Letrozole and Metformin Combined with Targeted Nursing Care on Ovarian Function, LH, and FSH in Infertile Patients with Polycystic Ovary Syndrome.
Polycystic ovary syndrome (PCOS) is a common gynecological endocrine disorder in women of gestational age and the most common cause of female infertility. This study aimed to investigate the effect of letrozole and metformin combined with targeted nursing on ovarian function, LH, and FSH in infertile patients with PCOS. A total of 72 infertile patients with PCOS were divided into the control group and combined group. The control group was given metformin tablets combined with targeted nursing therapy. The combined group was treated with letrozole on the basis of the control group. Ovarian function, LH, and FSH were compared between the two groups before and after treatment. After treatment, the total effective rate (94.44%) of the combined group was significantly higher than that of the control group (80.56%). After treatment, the levels of HbA1c, FINS, HOMA-IR, TG, and TC in the two groups were lower than those before treatment, and the HDL-C level was higher than that before treatment. The full-term delivery rate, ovulation rate, and normal menstrual rate in the combined group were higher than those in the control group. After treatment, the LH level in the combined group was lower than that in the control group, and the FSH level was higher than that in the control group. After 1 month, 3 months, and 5 months of treatment, RI, PI, and ovarian volume were decreased, and the endometrial thickness was increased in both groups. Also, after 5 months of treatment, the RI, PI, and ovarian volume in the combined group were significantly lower than those in the control group, and the endometrial thickness was significantly higher than that in the control group. There was no statistically significant difference in the incidence of adverse reactions between the two groups. The combination of letrozole and metformin with targeted nursing in the treatment of infertility patients with PCOS has better clinical effect and high safety. It can improve endometrial receptivity and ovarian reserve function and regulate LH and FSH levels.


[Effects of soil compactness stress on root activity and leaf photosynthesis of cucumber].
Responses of root activity and leaf photosynthesis to soil compactness stress were studied in cucumber plants grown in pots. Soil compaction was expressed by soil bulk density. There were three compactness treatments with soil bulk densities, 1.2, 1.4 and 1.6 g/cm(3). The results showed that when the soil compactness increased, the dry weight and activity of roots reduced (Fig. 1); the relative electrical conductivity and malondialdehyde (MDA) content of cucumber leaf (Fig. 2) increased; the soluble protein content decreased (Fig. 3); the activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) increased (Fig. 4); net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (E) and specific leaf weight (SLW) decreased, but intercellular CO(2) concentration (Ci) increased (Fig. 5). These results mean that high soil compaction brings stress to cucumber plants.


Symplasmic Isolation Contributes to Somatic Embryo Induction and Development in the Tree Fern Cyathea delgadii Sternb.
In this report, we describe studies on symplasmic communication and cellular rearrangement during direct somatic embryogenesis (SE) in the tree fern Cyathea delgadii. We analyzed changes in the symplasmic transport of low-molecular-weight fluorochromes, such as 8-hydroxypyrene-1,3,6-trisulfonic acid, trisodium salt (HPTS) and fluorescein (delivered to cells as fluorescein diacetate, FDA), within stipe explants and somatic embryos originating from single epidermal cells and developing during 16-d long culture. Induction of SE is preceded by a restriction in fluorochrome distribution between certain explant cells. Microscopic analysis showed a series of cellular changes like a decrease in vacuole size, increase in vacuole numbers, and increased density of cytoplasm and deposition of electron-dense material in cell walls that may be related with embryogenic transition. In somatic embryos, the limited symplasmic communication between cells was observed first in linear tri-cellular embryos. Further development of the fern embryo was associated with the formation of symplasmic domains corresponding to the four segments of the plant body. Using symplasmic tracers, we provided evidence that the changes in plasmodesmata permeability are corelated with somatic-to-embryogenic transition and somatic embryo development.


Scalable Colored Subambient Radiative Coolers Based on a Polymer-Tamm Photonic Structure.
Daytime radiative coolers cool objects below the air temperature without any electricity input, while most of them are limited by a silvery or whitish appearance. Colored daytime radiative coolers (CDRCs) with diverse colors, scalable manufacture, and subambient cooling have not been achieved. We introduce a polymer-Tamm photonic structure to enable a high infrared emittance and an engineered absorbed solar irradiance, governed by the quality factor (Q-factor). We theoretically determine the theoretical thresholds for subambient cooling through yellow, magenta, and cyan CDRCs. We experimentally fabricate and observe a temperature drop of 2.6-8.8 °C on average during the daytime and 4.0-4.4 °C during the nighttime. Furthermore, we demonstrate a scalable-manufactured magenta CDRC with a width of 60 cm and a length of 500 cm by a roll-to-roll deposition technique. This work provides guidelines for large-scale CDRCs and offers unprecedented opportunities for potential applications with energy-saving, aesthetic, and visual comfort demands.


The impact of the COVID-19 pandemic on physical activity and sleep among healthy adults: a systematic review and meta-analysis.
The COVID-19 pandemic has had a significant impact on physical and mental health, while physical activity and sleep are two important indicators of the impact that have been explored in recent studies. However, the results of studies with different measurement methods and populations with different levels of physical activity have been diverse in that physical activity and sleep are affected by the COVID-19 pandemic in some studies but not in others. Our study aimed to investigate the impact of the COVID-19 pandemic on physical activity and sleep and the role of measurement methods and populations on results. PubMed, Web of Science, and CNKI databases were used to search for related studies systematically. Study characteristics and data on physical activity and sleep were collected and analyzed from each included study. Standardized mean differences (SMDs) with 95% confidence intervals (CIs) were used to estimate pooled effect sizes. A total of 13 articles were included in the systematic review, 11 of which were included in the meta-analysis. We found that moderate-to-vigorous physical activity (MVPA) time was 0.33 (95% CI 0.07 to 0.59) and sleep quality was 0.37 (95% CI 0.21 to 0.53) decreased, while sleep duration was -0.24 (95% CI -0.28 to -0.20) increased during the lockdown; overall physical activity time had no significant difference (p = 0.07) during the lockdown. The "wearables" subgroup had no heterogeneity (p = 0.89, I2 = 0) in sleep duration, while MVPA time measured by subjective scales was not significantly changed. The "elite athletes" subgroup had lower heterogeneity (p = 0.69, I2 = 0) in sleep duration than general adults, while the results of sleep quality for population subgroups were significant and there was no heterogeneity within either. The COVID-19 pandemic had a significant impact on MVPA time, sleep duration, and sleep quality, instead of overall physical activity time among healthy adults. The results of MVPA time and sleep duration were greatly influenced by the measurement methods, and sleep behavior differed among populations with varying physical activity levels. Thus, when researching physical activity, especially MVPA time, should consider measurement methods, and more attention should be given to differences in populations when researching sleep behavior.


Glucocorticoid inhibition of granulocyte macrophage-colony-stimulating factor from T cells is independent of control by nuclear factor-kappaB and conserved lymphokine element 0.
Release of granulocyte macrophage-colony-stimulating factor (GM-CSF) from T cells is important in the differentiation, maturation, and survival of inflammatory cells. Here the induction of GM-CSF expression from T cells was dependent on transcription and translation and was prevented by dexamethasone. In primary human CD3(+) T cells, up to 3.3 kb of human GM-CSF promoter was strongly activated by PMA + PHA. Mutations in either the -85/-76 nuclear factor (NF)-kappaB site or the activator protein-1 region in the -54/-31 conserved lymphokine element 0 (CLE0) site substantially reduced promoter activity. Both GM-CSF promoter and NF-kappaB-dependent constructs were unresponsive to dexamethasone whereas the release of GM-CSF was potently repressed. Analysis of GM-CSF mRNA and protein expression at various time points and the effect of adding dexamethasone after the stimulus revealed the existence of potent mechanisms of inhibition acting at a translational level. The expression of tristetraproline and HuR, proteins that bind the AU-rich element in the GM-CSF 3'-untranslated region was unaffected by dexamethasone and overall AU-rich element binding activity was unaltered. Taken together our data support an important role for the NF-kappaB and CLE0 sites in the transcriptional control of GM-CSF expression in primary human T cells and suggest that post-transcriptional/translational mechanisms are key mediators of glucocorticoid-dependent repression.


A multistep oriented attachment kinetics: coarsening of ZnS nanoparticle in concentrated NaOH.
Crystal growth of ZnS nanoparticles during hydrothermal coarsened in 4 M NaOH occurs via a two-stage process. In the first stage, the primary particles grow into a size over hundred times of the original volume. The initial growth rate can be fitted by an asymptotic curve. High-resolution transmission electron microscope (HRTEM) data indicate that in this stage, crystal growth mainly occurs via a multistep crystallographically specific oriented attachment (OA). The higher the coarsening temperature, the earlier the first stage ends. In the second stage, an abrupt transition from asymptotic to square parabola growth kinetics occurs. The crystal growth data can be fitted by a standard Ostwald ripening (OR) model consistent with growth controlled by dissolution/precipitation of ions in solution. HRTEM data indicate that a minor amount of OA-based growth also occurs in the early period of the second stage. A new multistep OA kinetics model analogous to the reaction between molecules was proposed to illustrate the asymptotic growth in the first stage of coarsening. The effect of concentrated NaOH was discussed and proved to be the key that hindered the OR process, attributing to the almost exclusive pure OA-based growth of ZnS particles in the first stage.


Clinical anatomy of the inferior labial gland: a narrative review.
In this article we review the literature on the inferior labial gland from a clinical and anatomical perspective. Regardless of its importance in clinical practice, there are no medical literature that comprehensively reviewed the inferior labial gland. A database search using PubMed and Google Scholar was conducted. The following keywords were used in the search: "lower labial salivary gland", "lower labial gland", "inferior labial salivary gland", AND "inferior labial gland". The human labial glands are types of minor salivary gland that continuously secrete small amounts of mucous and serous substances to maintain oral health. The inferior labial glands are innervated by the inferior labial branch of the mental nerve, and the inferior labial branch of the facial artery is the main arterial supply to the lower lip. Although they only have an auxiliary role in saliva production compared to the major salivary glands, minor salivary glands provide a certain amount of lubrication in the oral cavity by the continuous outflow of saliva. The inferior labial gland not only promotes moisturization in the oral cavity but also secretes substances with antibacterial effects, which is important for the function of the oral cavity. A recent study showed that the rate of salivary secretion from the inferior labial glands does not change with age, and in some cases the inferior labial glands are used for diagnosing intractable diseases such as Sjogren's syndrome and cystic fibrosis. In addition, since the inferior labial glands themselves can be the site of cyst and/or neoplasia development, we should be careful to distinguish them from other diseases. Elucidation of the anatomy, physiology, and pathology of the inferior labial glands, is important for understanding human health and diseases.


New avenues to an old material: controlled nanoscale doping of germanium.
We review our recent research into n-type doping of Ge for nanoelectronics and integrated photonics. We demonstrate a doping method in ultra-high vacuum to achieve high electron concentrations in Ge while maintaining atomic-level control of the doping process. We integrated this doping technique with ultra-high vacuum scanning tunneling microscope lithography and femtosecond laser ablation micron-scale lithography, and demonstrated basic components of donor-based nanoelectronic circuitry such as wires and tunnel gaps. By repetition of controlled doping cycles we have shown that stacking of multiple Ge:P two-dimensional electron gases results in high electron densities in Ge (>10(20) cm(-3)). Because of the strong vertical electron confinement, closely stacked 2D layers - although interacting - maintain their individuality in terms of electron transport. These results bode well towards the realization of nanoscale 3D epitaxial circuits in Ge comprising stacked 2DEGs and/or atomic-scale Ge:P devices with confinement in more dimensions.


Sertoli-cell-specific knockout of connexin 43 leads to multiple alterations in testicular gene expression in prepubertal mice.
A significant decline in human male reproductive function has been reported for the past 20 years but the molecular mechanisms remain poorly understood. However, recent studies showed that the gap junction protein connexin-43 (CX43; also known as GJA1) might be involved. CX43 is the predominant testicular connexin (CX) in most species, including in humans. Alterations of its expression are associated with different forms of spermatogenic disorders and infertility. Men with impaired spermatogenesis often exhibit a reduction or loss of CX43 expression in germ cells (GCs) and Sertoli cells (SCs). Adult male transgenic mice with a conditional knockout (KO) of the Gja1 gene [referred to here as connexin-43 (Cx43)] in SCs (SCCx43KO) show a comparable testicular phenotype to humans and are infertile. To detect possible signaling pathways and molecular mechanisms leading to the testicular phenotype in adult SCCx43KO mice and to their failure to initiate spermatogenesis, the testicular gene expression of 8-day-old SCCx43KO and wild-type (WT) mice was compared. Microarray analysis revealed that 658 genes were significantly regulated in testes of SCCx43KO mice. Of these genes, 135 were upregulated, whereas 523 genes were downregulated. For selected genes the results of the microarray analysis were confirmed using quantitative real-time PCR and immunostaining. The majority of the downregulated genes are GC-specific and are essential for mitotic and meiotic progression of spermatogenesis, including Stra8, Dazl and members of the DM (dsx and map-3) gene family. Other altered genes can be associated with transcription, metabolism, cell migration and cytoskeleton organization. Our data show that deletion of Cx43 in SCs leads to multiple alterations of gene expression in prepubertal mice and primarily affects GCs. The candidate genes could represent helpful markers for investigators exploring human testicular biopsies from patients showing corresponding spermatogenic deficiencies and for studying the molecular mechanisms of human male sterility.


Human infections with Angiostrongylus cantonensis.
Angiostrongylus cantonensis is a parasite that causes eosinophilic meningitis and has been reported to be present on most Pacific islands. Rats are the principal host and several species of land snails the intermediate host. Important paratenic hosts are fresh water shrimp and fish. Modes of transmission include ingestion by man of raw fish, snails and fresh leafy vegetables contaminated by snail slime trails containing larvae. The parasitic worms are neurotropic in man, and the diagnosis should be considered in any adult or child, who presents, in endemic areas or areas with suitable intermediate hosts, with severe unrelenting headache, paresthesias, or a cranial nerve palsy. Eosinophils in the cerebral spinal fluid suggest the diagnosis. Simple analgesia is sufficient for mild cases. Treatment of those with severe symptoms remains controversial. Glucocorticoids, lumbar puncture to reduce intercranial pressure and antihelminthic agents have been used.


Confirmation of nasogastric tube placement by colorimetric indicator detection of carbon dioxide: a preliminary report.
Inadvertent insertion of nasogastric tubes into the trachea and distal airways is reported to range from 0.3% to 15% of insertions. Critically ill, mechanically ventilated patients are at a higher risk for such complications, some of which can be fatal. This preliminary prospective clinical investigation was designed to determine whether a colorimetric carbon dioxide (CO2) indicator device (Easy-Cap, Nellcor, Inc., Hayward, CA) attached to the proximal end of a small bore feeding tube (FT) would reliably discriminate between those tubes passed into the airways and those passed into the alimentary tract. Ten critically ill, mechanically ventilated trauma patients requiring a FT insertion were evaluated. An Easy-Cap device was adapted to the proximal port of each FT. Each patient had one tube inserted per the nasogastric route and then another through the endotracheal tube while the Easy-Cap was observed for color changes consistent with the presence of CO2. All transtracheal insertions showed immediate and unambiguous color changes consistent with the presence of CO2. None of the nasogastric insertions resulted in indicator color changes and all were confirmed with radiography (sensitivity 100%, specificity 100%, accuracy 100%). This preliminary report suggests colorimetric CO2 detection accurately and reliably identifies transtracheal FT insertion.


Methane-rich water induces cucumber adventitious rooting through heme oxygenase1/carbon monoxide and Ca(2+) pathways.
Methane-rich water triggered adventitious rooting by regulating heme oxygenase1/carbon monoxide and calcium pathways in cucumber explants. Heme oxygenase1/carbon monoxide (HO1/CO) and calcium (Ca(2+)) were reported as the downstream signals in auxin-induced cucumber adventitious root (AR) formation. Here, we observed that application of methane-rich water (MRW; 80% saturation) obviously induced AR formation in IAA-depleted cucumber explants. To address the universality, we checked adventitious rooting in soybean and mung bean explants, and found that MRW (50 and 10% saturation, respectively) exhibited the similar inducing results. To further determine if the HO1/CO system participated in MRW-induced adventitious rooting, MRW, HO1 inducer hemin, its activity inhibitor zinc protoporphyrin IX (ZnPP), and its catalytic by-products CO, bilirubin, and Fe(2+) were used to detect their effects on cucumber adventitious rooting in IAA-depleted explants. Subsequent results showed that MRW-induced adventitious rooting was blocked by ZnPP and further reversed by 20% saturation CO aqueous solution. However, the other two by-products of HO1, bilirubin and Fe(2+), failed to induce AR formation. Above responses were consistent with the MRW-induced increases of HO1 transcript and corresponding protein level. Further molecular evidence indicted that expression of marker genes, including auxin signaling-related genes and cell cycle regulatory genes, were modulated by MRW alone but blocked by the cotreatment with ZnPP, the latter of which could be significantly rescued by the addition of CO. By using the Ca(2+)-channel blocker and Ca(2+) chelator, the involvement of Ca(2+) pathway in MRW-induced adventitious rooting was also suggested. Together, our results indicate that MRW might serve as a stimulator of adventitious rooting, which was partially mediated by HO1/CO and Ca(2+) pathways.


Pre-natal exposure of mice to bisphenol A elicits an endometriosis-like phenotype in female offspring.
Endometriosis is a chronic gynecological disease characterized by the growth of endometrial tissue outside the uterine cavity. Exposure to endocrine disruptors during critical period of development causes long-lasting effects, being the genital system one of the targets. This study describes the effects on female genital system caused by developmental exposure to the endocrine-disrupting chemical bisphenol A (BPA) during pre- and peri-natal development in mice. To this end, timed pregnant Balb-C mice were treated from day 1 of gestation to 7 days after delivery with BPA (100, or 1000 microg/kg/day). After delivery, pups were held for 3 months; then, pelvic organs were analyzed in their entirety and livers of both pups and moms were studied for the presence of BPA. We found in the adipose tissue surrounding the genital tracts of a consistent number of treated animals, endometriosis-like structure with the presence of both glands and stroma and expressing both estrogen receptor and HOXA-10. Moreover, cystic ovaries, adenomatous hyperplasia with cystic endometrial hyperplasia and atypical hyperplasia were significantly more frequent in treated animals respect to the controls. Finally, BPA was found in the livers of exposed moms and female offspring. In conclusion, we describe for the first time an endometriosis-like phenotype in mice, elicited by pre-natal exposition to BPA. This observation may induce to thoroughly reconsider the pathogenesis and treatment of endometriosis, considering the high incidence of endometriosis and the problems caused by associated infertility.


Quantification of human mature frataxin protein expression in nonhuman primate hearts after gene therapy.
Deficiency in human mature frataxin (hFXN-M) protein is responsible for the devastating neurodegenerative and cardiodegenerative disease of Friedreich's ataxia (FRDA). It results primarily by epigenetic silencing the FXN gene due to up to 1400 GAA triplet repeats in intron 1 of both alleles of the gene; a subset of approximately 3% of FRDA patients have a mutation on one allele. FRDA patients die most commonly in their 30s from heart disease. Therefore, increasing expression of heart hFXN-M using gene therapy offers a way to prevent early mortality in FRDA. We used rhesus macaque monkeys to test the pharmacology of an adeno-associated virus (AAV)hu68.CB7.hFXN therapy. The advantage of using non-human primates for hFXN-M gene therapy studies is that hFXN-M and monkey FXN-M (mFXN-M) are 98.5% identical, which limits potential immunologic side-effects. However, this presented a formidable bioanalytical challenge in quantification of proteins with almost identical sequences. This was overcome by development of a species-specific quantitative mass spectrometry-based method, which revealed for the first time, robust transgene-specific human protein expression in monkey heart tissue. The dose response was non-linear resulting in a ten-fold increase in monkey heart hFXN-M protein expression with only a three-fold increase in dose of the vector.


Quantum and Molecular Mechanical (QM/MM) Monte Carlo Techniques for Modeling Condensed-Phase Reactions.
A recent review (Acc. Chem. Res. 2010, 43:142-151) examined our use and development of a combined quantum and molecular mechanical (QM/MM) technique for modelling organic and enzymatic reactions. Advances included the PDDG/PM3 semiempirical QM (SQM) method, computation of multi-dimensional potentials of mean force (PMF), incorporation of on-the-fly QM in Monte Carlo simulations, and a polynomial quadrature method for rapidly treating proton-transfer reactions. The current article serves as a follow up on our progress. Highlights include new reactions, alternative SQM methods, a polarizable OPLS force field, and novel solvent environments, e.g., "on water" and room temperature ionic liquids. The methodology is strikingly accurate across a wide range of condensed-phase and antibody-catalyzed reactions including substitution, decarboxylation, elimination, isomerization, and pericyclic classes. Comparisons are made to systems treated with continuum-based solvents and ab initio or density functional theory (DFT) methods. Overall, the QM/MM methodology provides detailed characterization of reaction paths, proper configurational sampling, several advantages over implicit solvent models, and a reasonable computational cost.


Characteristic changes of the S2/S1 difference FTIR spectrum induced by Ca2+ depletion and metal cation substitution in the photosynthetic oxygen-evolving complex.
Effects of Ca2+ depletion and substitution with other metal cations on the structure of the protein matrices of the oxygen-evolving complex (OEC) and their corresponding changes upon the S1 to S2 transition were examined using Fourier transform infrared (FTIR) spectroscopy. Ca2+ depletion and further supplementation with Li+, Na+, Mg2+, Ca2+, or Sr2+ did not significantly affect the typical vibrational features in the double difference S2/S1 spectrum, including the symmetric [1365(+)/1404(-) cm(-1)] and the asymmetric [1587(+)/1566(-) cm(-1)] stretching modes of the carboxylate ligand and the amide I and II modes of the backbone polypeptides. On the other hand, supplementation with K+, Rb+, Cs+, or Ba2+ significantly modified the S2/S1 spectrum, in which the carboxylate modes disappeared and the amide I and II modes were modified. Results indicate that the binding of metal cations that have ionic radii larger than that of Ca2+ to the Ca2+ site induces perturbations in the protein matrices in the vicinity of the Mn cluster to interrupt the characteristic structural and/or conformational changes upon the oxidation of the Mn cluster accompanied with the S1 to S2 transition. The spectrum was also altered by the supplementation of Cd2+, which has an ionic radius comparable to that of Ca2+. A single-pulse-induced S2/S1 difference spectrum revealed that bands that have been assigned to the vibrational modes for the Y(Z) tyrosine and the histidine ligand for the Mn cluster were not induced in the K+-supplemented membranes, although the histidine band is likely to be preserved in the Ca2+-depleted membranes. The Y(Z) band was considerably small in the double difference S2/S1 spectrum in the Ca2+-depleted and the cation-substituted membranes but distinctively present in the Sr2+- or Ca2+-replenished membranes. Furthermore, cation supplementation induced several new bands that disappeared following the Ca2+ replenishment. These results suggest that the proper organization of the hydrogen bond network within OEC for the water oxidation chemistry requires the Ca2+ ion and indicate that the role of Ca2+ is not purely structurally defined by the physical properties of the ion, such as valence and ionic radius. On the basis of these and other findings, we propose that Ca2+ is necessary for the formation of the hydrogen bond network that is involved in the reaction step of water oxidation.


Temporal-order discrimination for selected auditory and visual stimulus dimensions.
Thresholds for the discrimination of temporal order were determined for selected auditory and visual stimulus dimensions in 10 normal-adult volunteers. Auditory stimuli consisted of binary pure tones varying in frequency or sound pressure level, and visual stimuli consisted of binary geometric forms varying in size, orientation, or color. We determined the effect of psychophysical method and the reliability of performance across stimulus dimensions. Using a single-track adaptive procedure, Experiment 1 showed that temporal-order thresholds (TOTs) varied with stimulus dimension, being lowest for auditory frequency, intermediate for size, orientation, and auditory level, and longest for color. Test performance improved over sessions and the profile of thresholds across stimulus dimensions had a modest reliability. Experiment 2 used a double-interleaved adaptive procedure and TOTs were similarly ordered as in Experiment 1. However, TOTs were significantly lower for initially ascending versus descending tracks. With this method, the reliability of the profile across stimulus dimensions and tracks was relatively low. In Experiment 3, psychometric functions were obtained for each of the stimulus dimensions and thresholds were defined as the interpolated 70.7% correct point. The relative ordering of TOTs was similar to those obtained in the first two experiments. Non-monotonicities were found in some of the psychometric functions, with the most prominent being for the color dimension. A cross-experiment comparison of results demonstrates that TOTs and their reliability are significantly influenced by the psychophysical method. Taken together, these results support the notion that the temporal resolution of ordered stimuli involves perceptual mechanisms specific to a given sensory modality or submodality.


Involvement of the phosphatidylinositol 3-kinase/rac1 and cdc42 pathways in radial migration of cortical neurons.
During cortical development, newly generated neurons migrate radially toward their final positions. Although several candidate genes essential for this radial migration have been reported, the signaling pathways regulating it are largely unclear. Here we studied the role of phosphatidylinositol (PI) 3-kinase and its downstream signaling molecules in the radial migration of cortical neurons in vivo and in vitro. The expression of constitutively active and dominant-negative PI 3-kinases markedly inhibited radial migration. In the neocortical slice culture, a PI 3-kinase inhibitor suppressed the formation of GTP-bound Rac1 and Cdc42 and radial migration. Constitutively active and dominant-negative forms of Rac1 and Cdc42 but not Akt also significantly inhibited radial migration. In migrating neurons, wild-type Rac1 and Cdc42 showed different localizations; Rac1 localized to the plasma membrane and Cdc42 to the perinuclear region on the side of the leading processes. These results suggest that both the PI 3-kinase/Rac1 and Cdc42 pathways are involved in the radial migration of cortical neurons and that they have different roles.


Analysis of Mesorhizobium loti glycogen operon: effect of phosphoglucomutase (pgm) and glycogen synthase (g/gA) null mutants on nodulation of Lotus tenuis.
The phosphoglucomutase (pgm) gene codes for a key enzyme required for the formation of UDP-glucose and ADP-glucose, the sugar donors for the biosynthesis of glucose containing polysaccharides. A Mesorhizobium loti pgm null mutant obtained in this study contains an altered form of lipopolysaccharide (LPS), lacks exopolysaccharide (EPS), beta cyclic glucan, and glycogen and is unable to nodulate Lotus tenuis. The nonnodulating phenotype of the pgm mutant was not due to the absence of glycogen, since a glycogen synthase (glgA) null mutant effectively nodulates this legume. In M. loti, pgm is part of the glycogen metabolism gene cluster formed by GlgP (glycogen phosphorylase), glgB (glycogen branching), glgC (ADP-glucose pyrophosphorylase), glgA, pgm, and glgX (glycogen debranching). The genes are transcribed as a single transcript from glgP to at least pgm under the control of a strong promoter (promoter I) upstream of glgP. An alternative promoter (promoter II), mapping in a 154-bp DNA fragment spanning 85 bp upstream of the glgA start codon and the first 69 bp of the glgA coding region, controls the expression of glgA and pgm, independently of the rest of the upstream genes. Primer extension experiments showed that transcription starts 19 bp upstream of the glgA start codon.


Essential roles of G9a in cell proliferation and differentiation during tooth development.
Teeth develop through interactions between epithelial and mesenchymal tissues mediated by a signaling network comprised of growth factors and transcription factors. However, little is known about how epigenetic modifiers affect signaling pathways and thereby regulate tooth formation. We previously reported that the histone 3 lysine 9 (H3K9) methyltransferase (MTase) G9a is specifically enriched in the tooth mesenchyme during mouse development. In this study, we investigated the functions of G9a in tooth development using G9a conditional knockout (KO) mice. We used Sox9-Cre mice to delete G9a in the tooth mesenchyme because Sox9 is highly expressed in the mesenchyme derived from the cranial neural crest. Immunohistochemical analyses revealed that G9a expression was significantly decreased in the mesenchyme of Sox9-Cre;G9afl/fl (G9a cKO) mice compared with that in Sox9-Cre;G9a fl/+(control) mice. Protein levels of the G9a substrate H3K9me2 were also decreased in the tooth mesenchyme. G9a cKO mice showed smaller tooth germ after embryonic day (E) 16.5 and E17.5, but not at E15.5. The developing cusp tips, which were visible in control mice, were absent in G9a cKO mice at E17.5. At 3 weeks after birth, small first molars with smaller cusps and unseparated roots were formed. Organ culture of tooth germs derived from E15.5 cKO mouse embryos showed impaired tooth development, suggesting that tooth development per se is affected independently of skull development. BrdU labeling experiments revealed that the proliferation rates were decreased in the mesenchyme in G9a cKO mice at E17.5. In addition, the proliferation rates in the tooth inner enamel epithelium were also decreased. In situ hybridization revealed altered localization of genes associated with tooth development. In cKO mice, intensively localized expression of mRNAs encoding bone morphogenic protein (Bmp2 and Bmp4) was observed in the tooth mesenchyme at E17.5, similar to the expression patterns observed in control mice at E15.5. Localization of Shh and related signaling components, including Gli1, Ptch1, and Ptch2, in the tooth mesenchyme of cKO mice was generally similar to that at earlier stages in control mice. In addition, expression of Fgf3 and Fgf10 in the mesenchyme was decreased in G9a cKO mice at P0. Expression levels of Fgf9 and p21, both of which were expressed in the secondary enamel-knot, were also decreased. Thus, the expression of genes associated with tooth development was delayed in cKO mice. Our results suggest that H3K9MTase G9a regulates cell proliferation and timing of differentiation and that G9a expression in the tooth mesenchyme is required for proper tooth development.


Specificity of anti-lymphocyte antibodies in sera from patients with AIDS-related complex (ARC) and healthy homosexuals.
The presence and specificity of anti-lymphocyte antibodies (ALA) was investigated in sera from male homosexuals with AIDS-Related Complex (ARC) as well as healthy homosexuals. Individuals in the healthy homosexual group had no detectable antibodies to human immunodeficiency virus (HIV). Antibodies reactive with normal peripheral blood mononuclear cells were detected by Western blot analysis in sera from both groups of homosexuals. Of those individuals whose sera contained ALA, 71% of ARC patients and 83% of healthy homosexuals had antibodies recognizing a 73 kilodalton (kD) molecule. ALA present in ARC sera reacted with CD3+, CD4+ and CD8+ lymphocytes while little reactivity with B cells was observed. Our results indicate that ALA appear in homosexuals prior to HIV infection and are reactive primarily with T lymphocytes. A 73 kD structure associated with the T cell membrane is frequently the target for these antibodies.


Evolution of clinical trials for irritable bowel syndrome: issues in end points and study design.
Irritable bowel syndrome (IBS) involves a broad range of physiological and psychological alterations that may affect brain-gut dysregulation, gut function, visceral perception, and mucosal integrity and function. Despite advances in our understanding of basic neuroenteric mechanisms and the role of effectors and transmitters in the brain-gut axis, a reliable biologic marker of IBS has yet to be identified. IBS diagnosis and status depend entirely on an assessment of IBS signs and symptoms. This has made development of optimal end points and study design for evaluation of efficacy of IBS drugs a challenge. This article addresses three main topics: the evolution of primary end points for IBS clinical trials; a potential path forward for IBS end points in new clinical trials; and recommendations for the future development of patient-reported outcome (PRO) instruments for use in IBS clinical trials.


Effect of cannula position in the thoracic aorta with continuous left ventricular support: four-dimensional flow-sensitive magnetic resonance imaging in an in vitro model.
Left ventricular assist devices (LVADs) have become an important treatment option for heart failure patients. However, altered blood flow patterns are suspected to affect perfusion in the aorta or cause structural changes to the aortic root, leading to regurgitation and valve dysfunction or thrombus formation. The purpose of this study was to evaluate flow patterns in a realistic in vitro model system using four-dimensional flow-sensitive magnetic resonance imaging. A magnetic resonance compatible model system was developed consisting of an aorta connected to a VAD simulating the pulsatile flow of the native heart. An LVAD was connected to the aorta model via three different cannula positions. Flow patterns in the entire system as well as flow rates in predefined positions for reduced and zero cardiac output were evaluated. Cannula position influences flow patterns and flow rates in the entire thoracic aorta. For a residual cardiac output, a larger anastomosis and a decreased flow rate of the LAVD result in a higher flow rate and smaller retrograde flow in the ascending aorta when compared with a smaller anastomosis or a cannula position in the descending aorta. Pronounced flow turbulences in the aorta were observed for the cannula position in the descending aorta. In the setting of reduced cardiac output, as commonly observed in patients on LVAD therapy, a large anastomosis to the ascending aorta for the outflow cannula induces the least-adverse flow patterns in the aortic root. Our approach may aid in a better understanding of LVAD-induced flow-pattern changes. Optimization of the cannula position and anastomosis may help to prevent the progression of aortic valve-regurgitation and thrombus formation.


Factors affecting the decision to replace failed implants: a retrospective study.
The purpose of the present study was to explore the major factors that can affect the decision to replace failed implants. A retrospective cohort study was conducted on 194 patients who presented following dental implant failure during a 6-year period (2000 to 2006). The collected data included patient characteristics, failed implant characteristics, the anatomic status of the alveolar ridge after failure, and factors affecting the decision to avoid reimplantation. The study group included patients in whom the failed dental implants were replaced, whereas there was no reimplantation in the control group. Seventy-four patients (135 implants) made up the control group, and 120 patients (157 implants) made up the study group. The mean patient age was higher and the medical status was worse in the control group. The number of failed implants per patient was higher in the control group. The time between the diagnosis of failure and removal and between implant placement and removal were greater in the control group. The chances of a patient with minor bone loss undergoing reimplantation was 20 times greater (odds ratio, 20.4) than a patient with severe bone loss. The main patient-related reasons for avoiding reimplantation were the additional costs (27%), fear of additional pain (17.7%), and fear of a second failure (16.2%). The removal of a failing implant as soon as it is diagnosed as hopeless will improve the chances for reimplantation.


Changes in Bone Mineral Density, Body Composition, Vitamin D Status, and Mineral Metabolism in Urban HIV-Positive South African Women Over 12 Months.
Human immunodeficiency virus (HIV) infection and antiretroviral therapy (ART) are associated with bone loss and poor vitamin D status in white populations, though their relative roles are not known. No previous studies have examined longitudinal changes in areal bone mineral density (aBMD), measured by dual-energy X-ray absorptiometry (DXA), or in vitamin D status in HIV-positive African women. Of 247 premenopausal, urban, black African women from Soweto, South Africa, initially recruited, 187 underwent anthropometry, DXA scanning and blood and urine collections at both baseline and 12 months. Of these, 67 were HIV-negative throughout (Nref), 60 were HIV-positive with preserved CD4 counts at baseline (Ppres), and 60 were HIV-positive with low CD4 counts at baseline, eligible for ART by South African standards of care at the time (Plow). No participant had been exposed to ART at baseline. By 12 months, 51 Plow women had initiated ART, >85% of whom took combined tenofovir disoproxil fumarate (TDF), lamivudine, and efavirenz. By 12 months, Plow and Nref, but not Ppres, increased in body weight and fat mass (group-by-timepoint p ≤ 0.001, p = 0.002, respectively). Plow had significant decreases in aBMD of 2% to 3%, before and after size adjustment, at the femoral neck (p ≤ 0.002) and lumbar spine (p ≤ 0.001), despite significant weight gain. These decreases were associated with increased bone turnover but there were no significant differences or changes over time in vitamin D status, serum phosphate concentrations, or renal phosphate handling. Excluding data from nine Plow women unexposed to ART and 11 Ppres women who had initiated ART accentuated these findings, suggesting the bone loss in Plow was related to ART exposure. This is the first study describing DXA-defined bone loss in HIV-positive Sub-Saharan African women in association with ART. Further work is required to establish if bone loss continues with ongoing ART and, if so, whether this results in increased fracture rates. © 2017 American Society for Bone and Mineral Research.


The immunosuppressive effect of vincristine on allostimulatory potential of human dendritic cells interferes with their function and survival.
Mature dendritic cells (mDC) are professional and potent antigen presenting cells required for initiation of primary immune responses. In the present study, we have investigated the effect of vincristine on the T cell allostimulatory potential of human monocyte-derived mDC in allogeneic mixed lymphocyte reaction. Using T lymphocytes as responding cells and mDC as stimulating cells, our data indicate that incubation of DC with vincristine decreased the accessory potency dose dependently and resulted in a subsequent inhibition of T cell proliferative responses. Treatment of mDC with vincristine also led to the alteration of their capacity to produce IL-12 but enhanced their production of IL-10. Using flow cytometry, we demonstrated that vincristine had no effect on mDC phenotype (CD83, CD40, CD86, CD58, CD54) but promoted apoptotic cell death of these cells as revealed by PI and annexin-V. These findings suggest that DC may be potential targets of cytotoxic drugs and point out the possible impairment of the immunocompetence of these cells following chemotherapy.


Coronary artery bypass on survival in patients with moderate ischemic mitral regurgitation: meta-analysis.
The objective of the study was to systematically evaluate the effect of coronary artery bypass grafting (CABG) or CABG combined with mitral valve surgery (cMVS) on post-operative survival in patients with moderate ischemic mitral valve regurgitation. Databases including PubMed, Web of Science, COCHRANE LIBRARY, WanFang Data, and CNKI Data were searched from inception to January 2020. According to the inclusion criterion, relevant articles were screened. After that we extracted data, assessed quality, and performed meta-analysis using RevMan 5.2. A total of 4 randomized controlled trial and 14 retrospective study involving 4476 patients were included in the study. The CABG group was 2278 and the cMVS group was 1698. The results of meta-analysis showed that compared with CABG group, there were no statistically significant differences in the recent mortality (odds ratio [OR] = 0.88, p = 0.62), 1-year survival (OR = 1.03, p = 0.82), 1-year survival (OR = 1.07, p = 0.62), and long-term survival (OR = 0.95, p = 0.61) of the cMVS group. Current evidence indicates that patients in the cMVS group did not benefit from CABG group in survival after surgery.


[Environmental factors in juvenile delinquency].
Experimental data are cited for the proposition that the complicated aspects of juvenile delinquency can only be understood and explained by adopting a simultaneous psychological and sociological approach. It is also shown that the manifestations of delinquency, though not its actual presence, may be influenced by sex and a depressed city or rural background. Environmental factors serving as stimulating features and hereditary (i.e. predisposing) factors undoubtedly contribute to the formation of the Ego. The former, however, are elaborated by their receipient and are not sufficient to explain a certain type of behaviour. The influence of cultural models should not be underestimated. These, where predominant, tend to render normative what may be considered as deviant.


15-deoxyspergualin: from cytostasis to immunosuppression.
This article gives a survey on the pharmacological action of the new polyamine compound 15-Deoxyspergualin (15-DS). 15-DS is a derivative of the antitumor antibiotic spergualin which has been isolated by Prof. Umezawa's group in Japan. Besides its antitumor activity 15-DS was shown to possess immunosuppressive properties making the substance suitable for the application in organ transplantation and autoimmune disorders. Due to its immunopharmacological mode of action known hitherto 15-DS clearly differs from other immunosuppressants as e.g. Cyclosporin A.


The influence of narrow-leafed lupin seed fermentation on their chemical composition and ileal digestibility and microbiota in growing pigs.
The aims of this study were to provide a controlled fermentation process of blue lupin seeds (Lupinus angustifolius, cv. Neptun), monitor the changes in seed composition and determine the influence of the fermentation on the apparent ileal digestibility (AID) of crude protein (CP) and amino acids in growing pigs, compared with raw lupin seeds. The fermentation with bacteria and yeast was conducted for 24 h at 25ºC under aerobic conditions. Seed fermentation increased the contents of CP, fibre, fat and ash and most of the analysed amino acids but reduced the levels of the nitrogen-free extractives. Furthermore, fermentation decreased the contents of raffinose family oligosaccharides and phytic acids but increased the alkaloid content. The AID was estimated on three barrows (mean initial body weight 25 kg), surgically fitted with a T-cannula in the distal ileum. The pigs received three diets, each for 6 d, within three experimental periods (3 × 3 Latin Square design). The diets contained soybean meal (Group SBM), raw lupin seeds (Group RL) or fermented lupin seeds (Group FL) as solely protein sources. Fermentation had a positive impact on the AID of CP and methionine, cysteine, isoleucine, leucine, phenylalanine and valine (p < 0.05). Feeding raw or fermented lupin seeds did not affect the microbial status of the ileal digesta. Moreover, ammonia content in the caecal digesta did not differ between Groups RL and FL, although it was significantly higher than in Group SBM. It can be concluded that the fermentation process modified the chemical composition of nutrients in seeds, which can influence the digestibility and utilisation of the fermentation product in animal diets compared to raw seeds.


Effects of low doses of UV-B radiation supplementation on tuber quality in purple potato (Solanum tuberosum L.).
UV-B is an important environmental factor that differentially affects plant growth and secondary metabolites. However, our knowledge regarding the physiological and biochemical changes in under-ground plant organs responded to UV-B treatment remains limited. In this study, we investigated potato plant (Solanum tuberosum L.) and tuber responses to short-term supplemental UV-B exposure performed during tuber development. Our results indicated that the supplemental UV-B radiation with relative low dose had no obvious adverse impact on plant growth or tuber production. Nutritional composition analyses of tubers revealed that the contents of starch, soluble sugars, and proteins were significantly increased under lower UV-B radiation relative to controls. Similarly, low dose of UV-B treatment promoted the health-promoting compounds, including anthocyanin, phenols, and flavonoids in tubers. Moreover, higher activities of antioxidant enzymes were significantly induced in tubers in response to lower UV-B radiation. These findings suggest that short-term UV-B radiation supplementation at relative low doses can improve the tuber quality in potato plants.


Connexin43 and connexin45 form heteromeric gap junction channels in which individual components determine permeability and regulation.
Two gap junction proteins, connexin43 (Cx43) and connexin45 (Cx45), are coexpressed in many cardiac and other cells. Homomeric channels formed by these proteins differ in unitary conductance, permeability, and regulation. We sought to determine the ability of Cx43 and Cx45 to oligomerize with each other to form heteromeric gap junction channels and to determine the functional and regulatory properties of these heteromeric channels. HeLa cells were transfected with Cx45 or (His)(6)-tagged Cx43 or sequentially transfected with both connexins. Immunoblots verified production of the transfected connexins, and immunofluorescence demonstrated that they were colocalized in the HeLa-Cx43(His)(6)/Cx45 cells. Connexons were solubilized from HeLa-Cx43(His)(6)/Cx45 cells by using Triton X-100 and were applied to a Ni(2+)-NTA column, which binds the His(6) sequence. Cx45 was coeluted from the column with Cx43(His)(6), demonstrating that some hemichannels contain both connexins. Single-channel recordings showed that the HeLa-Cx43(His)(6)/Cx45 cells exhibited single-channel conductances that were not observed in cells expressing either connexin alone. Dye-coupling experiments showed that HeLa-Cx43(His)(6) cells readily passed Lucifer yellow and N-(2-aminoethyl)biotinamide hydrochloride (neurobiotin); in contrast, HeLa-Cx45 and HeLa-Cx43(His)(6)/Cx45 cells showed extensive intercellular passage of neurobiotin but little coupling with Lucifer yellow. Treatment with the protein kinase C activator 12-O-tetradecanoylphorbol 13-acetate reduced junctional conductance in cells expressing Cx43, Cx45, or both connexins, but it reduced the extent of neurobiotin transfer only in HeLa-Cx43(His)(6) and HeLa-Cx43(His)(6)/Cx45 cells but not in the HeLa-Cx45 cells. Thus, biochemical and electrophysiological evidence suggests that Cx43 and Cx45 extensively mix to form heteromeric channels; however, individual connexin components dominate aspects of the physiological behavior of these channels.


[Severe astigmatism following transfixing keratoplasty: how to attempt to reduce it].
Surgical treatment of astigmatism has been practised since the nineteenth century. Ophthalmologists' attitude has to change with respect to the problem of astigmatism after penetrating keratoplasty. A high astigmatism must no longer be considered as a fatality, particularly in young patients, except in special circumstances to be considered case by case. Rather than waiting for corneal healing, and operating only some time after ablation of the edge-to-edge suture by practising for example a wedge resection, it appears logical to act sooner to benefit from the temporary flexibility of a recent scar. Initially, 5 or 6 very tight sutures should be inserted in the axis of the flatter meridian as a routine procedure. Overcorrection must be as high as the astigmatism to be corrected; the total correction achieved will be reduced by about 50% in the following weeks and will then become stable. The sutures have to be kept in position until the corneal scar is completely healed. Such a method has the drawback of remaining approximate, but this is also the case with other operations proposed at a later stage. It has the advantage of being simple and without risk of lasting overcorrection. This study was essentially a prospective and preliminary one, and analysis of results with more cases will determine whether this method is effective.


Small cell neuroendocrine carcinoma of the uterine cervix presenting with syndrome of inappropriate antidiuretic hormone secretion.
Small cell carcinoma of the uterine cervix is rare. It is estimated that 10% of patients with small-cell lung cancer have syndrome of inappropriate secretion of antidiuretic hormone (SIADH) and hyponatremia has been reported to be significantly associated with a poor prognosis. A proportion of small cell carcinoma of the uterine cervix exhibit neuroendocrine characteristics as revealed by immunohistochemistry, However, cases presenting typical symptoms due to SIADH are extremely rare. This report of the SIADH of the uterine cervix is a rare case in the small cell carcinoma of the cervix presenting with tumor-associated paraneoplastic syndrome.


Successive surveys of an expert panel: research in decision-making with health visitors.
A United Kingdom expert panel was consulted to develop a self-completion instrument for research on decision making in health visiting practice. The use of a Delphi-type technique demonstrated how a stand-alone postal survey was an insufficient measure for health visitors, requiring a supplementary follow-up method to study types of casework. A review of literature raised questions about the significance of intuitive thinking and reasoning in health visitor practice, in relation to measuring professional service and individual performance.


An in-situ photocrosslinking microfluidic technique to generate non-spherical, cytocompatible, degradable, monodisperse alginate microgels for chondrocyte encapsulation.
Alginate microgels are widely generated by ionic crosslinking methods, but this method has limitations in controlling the microgel degradation and generating non-spherical microgels. By employing oxidized methacrylated alginate (OMA) that is degradable and photocrosslinkable, we have successfully photocrosslinked monodisperse OMA microgels and demonstrated the feasibility to generate discoid alginate microgels. However, several technical issues obstructed our opto-microfluidic method from being a useful technique. Here, we further characterized and optimized this method. Monodisperse discoid OMA microgels with good shape consistency were, for the first time, generated. The curability of OMA microgels was characterized as the macromer concentration varied from 2% to 10%, and the minimum required photoinitiator (VA-086) concentrations were determined. The effects of crosslinking density and the presence of ions in the storage solution on swelling of OMA hydrogels were identified to give insights into accurate controlling of the microgel size. A much quicker degradation rate (within three weeks) compared to ionically crosslinked alginate hydrogels was indirectly identified by quantifying the elastic modulus using atomic force microscopy. The viability of encapsulated chondrocytes in OMA microgels formed by this method was higher than those from other existing methods, demonstrating its favorable cytocompatibility. It was found that the oxygen tension played a critical role in both the curability of microgels and the cytocompatibility of this technique. We also summarize common practical issues and provide related solutions and/or operational suggestions. By this method, OMA microgels are expected to be valuable alternatives to traditional ionically crosslinked alginate microgels in drug delivery, tissue engineering, and single cell analysis areas due to their multiple favorable properties.


Differential cytokine production and Toll-like receptor signaling pathways by Candida albicans blastoconidia and hyphae.
Toll-like receptors (TLR) are crucial for an efficient antifungal defense. We investigated the differential recognition of blastoconidia and hyphae of Candida albicans by TLRs. In contrast to Candida blastoconidia, which stimulated large amounts of gamma interferon (IFN-gamma), the tissue-invasive Candida hyphae did not stimulate any IFN-gamma by human peripheral blood mononuclear cells (PBMC) or murine splenic lymphocytes. After stimulation with blastoconidia, the production of IFN-gamma was TLR4 dependent, as shown by the significantly decreased IFN-gamma production in anti-TLR4-treated PBMC and in splenic lymphocytes from TLR4-defective ScCr mice. In addition, peritoneal macrophages from ScCr mice produced less tumor necrosis factor alpha (TNF-alpha) than macrophages of control mice did when stimulated with Candida blastoconidia, but not with hyphae, indicating that TLR4-mediated signals are lost during hyphal germination. In contrast, macrophages from TLR2 knockout mice had a decreased production of TNF-alpha in response to both Candida blastoconidia and hyphae. Candida hyphae stimulated production of interleukin-10 through TLR2-dependent mechanisms. In conclusion, TLR4 mediates proinflammatory cytokine induction after Candida stimulation, whereas Candida recognition by TLR2 leads mainly to anti-inflammatory cytokine release. TLR4-mediated proinflammatory signals are lost during germination of Candida blastoconidia into hyphae. Phenotypic switching during germination may be an important escape mechanism of C. albicans, resulting in counteracting host defense.


Xylopiana A, a Dimeric Guaiane with a Case-Shaped Core from Xylopia vielana: Structural Elucidation and Biomimetic Conversion.
Xylopiana A (1), a dimeric guaiane with an unprecedented pentacyclo[5.2.1.01,2.04,5'.05,4']decane-3,2'-dione core, and three biosynthetically related intermediates, compounds 2-4, were isolated from the leaves of Xylopia vielana. Their structures and absolute configurations were determined by a combination of spectroscopic data, X-ray crystallography, electronic circular dichroism calculations, and chemical conversion. The structure of known vielanin A was revised to be compound 3. Compound 4 exerted a 3.7-fold potentiation effect on doxorubicin susceptibility at the tested concentration of 10 μM.


IL-10 synergizes with IL-4 and IL-13 in inhibiting lysosomal enzyme secretion by human monocytes and lamina propria mononuclear cells from patients with inflammatory bowel disease.
Tissue injury and inflammation in inflammatory bowel disease (IBD) are associated with enhanced monocytic lysosomal enzyme release. In this study, peripheral monocytes and lamina propria mononuclear cells (LPMNC) were isolated from IBD patients and normal controls. Cells were stimulated with lipopolysaccharide after treatment with IL-13, IL-4, and IL-10, and enzyme secretion was assessed by using the corresponding p-nitrophenyl glycosides as substrates. Molecular forms of cathepsin D were examined to describe the mode of enzyme release. IL-10 and IL-4 strongly down-regulate enzyme secretion in IBD monocytes. IBD monocytes showed a diminished responsiveness to the inhibitory effect of IL-13. Impaired monocyte response was not found with combinations of IL-13 and IL-10 or IL-4 and IL-10. LPMNC from involved IBD mucosa showed significantly higher enzyme secretion compared with LPMNC from noninvolved IBD mucosa but responded inefficiently to either IL-4, IL-13, or IL-10 alone. However, combined treatment with IL-10 and IL-4 or IL-10 and IL-13 strongly suppressed enzyme release by these cells. Both the precursor and mature forms of cathepsin D were elevated in IBD patients. While IL-13 reduced mainly the precursor form, the effect of IL-4 and IL-10 concerns both the precursor and mature form of cathepsin D. Our results favor the potent clinical utility of combined treatment, thus improving chances of developing effective treatments for human IBD.


Long-term efficacy and safety of lamotrigine for all types of bipolar disorder.
We investigated whether the long-term efficacy and safety of lamotrigine (LTG) for bipolar disorder (BP) differs between disease types (BP-I, BP-II, or BP not otherwise specified [BP-NOS]), and the efficacy of the concomitant use of antidepressants (ADs). For >1 year, we observed 445 outpatients with BP (diagnosed by DSM-IV criteria) who initiated LTG treatment between July 1 and October 31, 2011, using the Himorogi Self-rating Depression (HSDS) and Anxiety Scales and the Clinical Global Impression-Improvement scale and also recorded adverse events. Treatment efficacy was observed at week 4, with the improved HSDS scores sustained until week 52 for all types of BP; 50% of the patients with any type of BP could be treated with LTG for 1 year, whereas ~40% could be treated for >1.5 years. However, 25% of the patients were withdrawn within the first 4 weeks. The overall incidence of adverse events was 22.9% (104/455): 34.1% (14/41) for BP-I, 22.7% (15/66) for BP-II, and 22.2% (75/338) for BP-NOS. The most common adverse event was skin rash: 22.0% for BP-I, 16.7% for BP-II, and 12.1% for BP-NOS. There was no control group. Data were collected retrospectively. With careful and adequate titration, long-term treatment with LTG is possible for any type of BP, with BP-NOS patients, the largest population in clinical practice, responding particularly well. Symptoms can improve with or without ADs. Large-scale prospective studies of the efficacy of ADs in bipolar treatment are warranted.


Soft tissue sarcomas in adolescents and young adults: a comparison with their paediatric and adult counterparts.
Survival outcomes for adolescent and young adult patients with soft tissue sarcomas lag behind those of children diagnosed with histologically similar tumours. To help understand these differences in outcomes, we discuss the following issues with regard to the management of these patients with soft tissue sarcomas: delays in diagnosis, trial availability and participation, aspects of the organisation of care (with an emphasis on age-specific needs), national centralisation of sarcoma care, international consortia, and factors related to tumour biology. Improved understanding of the causes of the survival gap between adolescents and young adults with sarcomas will help drive new initiatives to improve final health outcomes in these populations. In this Review, we specifically focus on embryonal and alveolar rhabdomyosarcoma, synovial sarcoma, and adult soft tissue sarcomas diagnosed in adolescents and young adults, and discuss the age-specific needs of these patients.


Anti-vascular endothelial growth factor therapy in the era of personalized medicine.
To review the role of anti-vascular endothelial growth factor (anti-VEGF) therapies in the personalized medicine era. We searched PubMed for prospective clinical trials published through October 2012 of anti-VEGF agents approved by the U.S. Food and Drug Administration or the European Medicines Agency. The use of anti-VEGF drugs as single agents or in combination with other targeted or cytotoxic agents was associated with improved response rates and progression-free survival. Anti-VEGF therapy exerts its action by blocking tumor vessel formation and, thus, proliferation. Some investigators demonstrated modest to no improvement in overall survival, although the maintenance of anti-VEGF therapy beyond progression was shown to result in longer overall survival. The use of anti-VEGF therapy was associated with adverse events (i.e., thromboembolism, hemorrhage, myocardial infarction, and hypertension) and transformation to a more invasive phenotype. The development of multikinase targeting agents that include anti-VEGF properties warrants further investigation. The role of anti-VEGF therapy is evolving in the era of personalized medicine, and its use needs to be reassessed in tumor types with effective FDA-approved targeted agents, especially in light of its relatively high cost.


Effect of femoral head size and abductors on dislocation after revision THA.
Dislocation is a common complication after revision total hip arthroplasty, particularly if the abductor mechanism is deficient. Use of a large femoral head may reduce the incidence of dislocation. However, it is not clear if the large femoral head is effective in controlling dislocation when the abductor mechanism is deficient. We separated 230 patients who underwent revision total hip arthroplasty into four groups: Group 1 (159 patients) had an intact abductor mechanism and a 28-mm femoral head, Group 2 (20 patients) had an absent abductor mechanism (trochanteric nonunion or complete segmental proximal femoral bone loss) and a 28-mm femoral head, Group 3 (42 patients) had an intact abductor mechanism and a 36-mm femoral head, and Group 4 (nine patients) had an absent abductor mechanism and a 36-mm femoral head. The minimum followup was 6 months (mean, 27 months; range, 6 months to 7 years). A 36-mm head was associated with a lower dislocation rate after revision total hip arthroplasty than a 28-mm head with an intact abductor mechanism; the rate of dislocation was 12.7% for Group 1, 40.0% for Group 2, 0% for Group 3, and 33.3% for Group 4. However, the use of a large-diameter head does not reduce the rate of dislocation if the abductor mechanism is absent.


Evaluating shock absorption behavior of small-sized systems under programmable electric field.
A simple ball-drop impact tester is developed for studying the dynamic response of hierarchical, complex, small-sized systems and materials. The developed algorithm and set-up have provisions for applying programmable potential difference along the height of a test specimen during an impact loading; this enables us to conduct experiments on various materials and smart structures whose mechanical behavior is sensitive to electric field. The software-hardware system allows not only acquisition of dynamic force-time data at very fast sampling rate (up to 2 × 10(6) samples/s), but also application of a pre-set potential difference (up to ±10 V) across a test specimen for a duration determined by feedback from the force-time data. We illustrate the functioning of the set-up by studying the effect of electric field on the energy absorption capability of carbon nanotube foams of 5 × 5 × 1.2 mm(3) size under impact conditions.


Marital quality and congruent drinking.
This research considered whether changes in marital quality over the early years of marriage were related to patterns of alcohol use among three groups of couples: congruent nondrinkers, congruent drinkers who usually drank with their spouses and congruent drinkers who usually drank apart from their spouses. Newlywed couples (N = 418) were assessed for marital satisfaction and drinking behaviors and then were reassessed at their first and second anniversaries. Cross-sectional analyses compared couples at each assessment and multilevel modeling assessed changes in marital satisfaction over time. At each assessment, husbands and wives who usually drank with their partners reported greater levels of marital satisfaction. Over time, marital satisfaction declined for both husbands and wives. When we assessed changes in marital quality based on the three groups, husbands in each group experienced similar declines in marital quality. Among wives, however, the rate of decline was not the same. Although wives in the nondrinking group and wives who usually drank with their husbands had similar initial marital satisfaction, the nondrinkers experienced a greater decline in marital satisfaction than the wives who drank with their husbands. The rate of change for the wives in the nondrinking group was quite similar to wives who more often drank apart from their spouses. These findings suggest that alcohol use may be a part of the couple's socializing and may increase interaction, thereby increasing marital satisfaction.


Design novel dual agonists for treating type-2 diabetes by targeting peroxisome proliferator-activated receptors with core hopping approach.
Owing to their unique functions in regulating glucose, lipid and cholesterol metabolism, PPARs (peroxisome proliferator-activated receptors) have drawn special attention for developing drugs to treat type-2 diabetes. By combining the lipid benefit of PPAR-alpha agonists (such as fibrates) with the glycemic advantages of the PPAR-gamma agonists (such as thiazolidinediones), the dual PPAR agonists approach can both improve the metabolic effects and minimize the side effects caused by either agent alone, and hence has become a promising strategy for designing effective drugs against type-2 diabetes. In this study, by means of the powerful "core hopping" and "glide docking" techniques, a novel class of PPAR dual agonists was discovered based on the compound GW409544, a well-known dual agonist for both PPAR-alpha and PPAR-gamma modified from the farglitazar structure. It was observed by molecular dynamics simulations that these novel agonists not only possessed the same function as GW409544 did in activating PPAR-alpha and PPAR-gamma, but also had more favorable conformation for binding to the two receptors. It was further validated by the outcomes of their ADME (absorption, distribution, metabolism, and excretion) predictions that the new agonists hold high potential to become drug candidates. Or at the very least, the findings reported here may stimulate new strategy or provide useful insights for discovering more effective dual agonists for treating type-2 diabetes. Since the "core hopping" technique allows for rapidly screening novel cores to help overcome unwanted properties by generating new lead compounds with improved core properties, it has not escaped our notice that the current strategy along with the corresponding computational procedures can also be utilized to find novel and more effective drugs for treating other illnesses.


Deep Margin Elevation: Next-Level Adhesive Dentistry to Avoid Surgical Crown Lengthening.
Surgical crown lengthening (SCL) is the treatment of choice to ensure healthy tissues when subgingival tooth defects encroach on the biologic width.1 However, restorative techniques employing adhesive dentistry may provide viable alternatives to or complement SCL in select cases without the repercussions of surgical bone removal and open interproximal spaces. One such clinical technique, deep margin elevation (DME), employs a direct restoration to relocate the cervical margin of small subgingival defects of posterior teeth supragingivally. The "elevated" margin simplifies impression-making and bonding of indirect restorations, especially inlays and onlays. Recent scientific systematic literature reviews indicate favorable clinical outcomes and suggest that DME restorations made with scrupulous care have high success rates and are compatible with periodontal health. Optimal working field isolation, meticulous placement of matrices, proper bonding and buildup procedure, as well as regular maintenance and follow-up are essential for success. This article provides an overview of this approach.


Hydrogeochemistry of boron in borewell water of Durgapur coal mine area, District Chandrapur, Maharashtra, India.
Two formations, Barakar and Talchir belonging to Lower Gondwana sequence form aquifer in Durgapur area. Groundwater of these formations sampled for boron investigation in two seasons, Post-monsoon and Pre-monsoon. The water quality differed from aquifer to aquifer, mainly with respect to conductivity and B contents. Talchir groundwater shows greater conductivity and greater B content than Barakar groundwater. The maximum concentration of B was found in Talchir pre-monsoon groundwater. Greater B in Talchir groundwater is attributed to glacio-marine environment of sediment deposition. The correlation of B with major ions also varies. It shows positive correlation with conductivity, Na, HCO3 and SO4 in Barakar groundwater and with SO4 in Talchir groundwater. Seasonal variation in concentration of B exists but not appreciable.


Circularization, photomechanical switching, and a supercoiling transition of actin filaments.
We confine actin filaments onto a 2D surface using depletion interactions and show that this significantly increases the probability of intramolecular circularization. Quantitative analysis reveals that the resulting semiflexible rings fluctuate significantly less then their linear counterparts with equal stiffness-an effect induced by the constraint of circular geometry. When exposed to fluorescence excitation light, rhodamine-phalloidin-labeled filaments undergo a change in their natural twist. This photomechanical transition induces a localized small-wavelength supercoiling transition of absorbed actin rings. Upon completion of the photoinduced reaction, the twist of neighboring monomers in an actin filament changes by approximately 0.26 degrees .


Arterial Tortuosity and Its Correlation with White Matter Hyperintensities in Acute Ischemic Stroke.
The association between arterial tortuosity and acute ischemic stroke (AIS) has been reported, but showing inconsistent results. We hypothesized that tortuosity of extra- and intracranial large arteries might be higher in AIS patients. Furthermore, we explored the correlation between artery tortuosity and white matter hyperintensity (WMH) severity in AIS patients. 166 AIS patients identified as large artery atherosclerosis, and 83 control subjects were enrolled. All subjects received three-dimensional computed tomography angiography (CTA). Arterial tortuosity was evaluated using the tortuosity index. WMHs were evaluated using magnetic resonance imaging in all AIS patients. AIS patients showed significantly increased arterial tortuosity index relative to controls, including left carotid artery (CA) (p = 0.001), right CA (p < 0.001), left common carotid artery (CCA) (p < 0.001), right CCA (p < 0.001), left internal carotid artery (p = 0.001), right internal carotid artery (p = 0.01), left extracranial internal carotid artery (EICA) (p < 0.001), right EICA (p = 0.01), and vertebral artery dominance (VAD) (p = 0.001). The tortuosity of all above arteries was associated with the presence of AIS. AIS patients with moderate or severe WMHs had a higher tortuosity index in left CA (p = 0.005), left CCA (p = 0.003), left EICA (p = 0.07), and VAD (p = 0.001). In addition, the tortuosity of left EICA and VAD was associated with WMH severity in AIS patients. Increased extra- and intracranial large arteries tortuosity is associated with AIS. The tortuosity of left carotid artery system and vertebral artery may be the independent risk factors for WMH severity in AIS patients. Clinical Trial Registration. This trial is registered with NCT03122002 (http://www.clinicaltrials.gov).


Quality of life and Economic Aspects of Community Support Programs for Caregivers of Dementia Patients.
This economic analysis of community programs for caregivers of dementia patients first reviews the clinical data on caregiver stress and the effectiveness of a variety of intervention programs. The authors then discuss issues relevant to the measurement of the true costs of specific community support programs and their impact on the quality of life of caregivers. Using cost data derived from other published studies, the authors conducted a cost-utility analysis using incremental costs per quality-adjusted life-year (QALY) gained. A sensitivity analysis of cost: utility ratios reveals a range from $3,333 (1988 US. dollars) to $28,571 per QALY gained. Although this is comparable to other health care programs, community support programs for caregivers of dementia patients present special challenges for evaluation of outcome.


IgG subclass responses to pigeon intestinal mucin are related to development of pigeon fanciers' lung.
Pigeon fanciers' lung (PFL) is a form of extrinsic allergic alveolitis. Affected individuals produce antibodies to various pigeon antigens, and the resulting immune complexes are thought to initiate the disease. However, high antibody titres also occur in some asymptomatic individuals. Previously attention has focused on protein antigens, but we have recently identified pigeon intestinal mucin as a novel antigen in PFL. To determine the relationship between IgG subclass antibodies to pigeon intestinal mucin and the development of pigeon fanciers' lung. Sera were collected from 250 pigeon fanciers, who also completed a clinical questionnaire. Sera were screened for precipitating antibodies to pigeon serum and droppings. Individuals with symptoms and precipitating antibodies were considered to have classical PFL. Serum IgG and IgG subclass antibodies to pigeon intestinal mucin and pigeon serum proteins were investigated by quantitative enzyme-linked immunosorbent assay (ELISA). Very high titres of IgG antibodies against pigeon mucin were found in all precipitin-positive individuals. A strong positive correlation was seen between titres of antibodies to mucin and to serum proteins, but this was not due to crossreactivity. No significant differences in IgG titres to either mucin or pigeon serum proteins were found between individuals with PFL and asymptomatic precipitin positive fanciers. IgG1 and IgG2 were the major subclasses of anti-mucin, with lower titres of IgG3. Patients with PFL had significantly higher titres of IgG1 to mucin than asymptomatic, precipitin-positive individuals. In contrast, no significant differences were seen between PFL and asymptomatic precipitin-positive sera with respect to the subclass titres against pigeon serum proteins. The high titres of anti-mucin IgG in sera of all individuals with PFL, together with the finding that high IgG1 titres to mucin are associated with the development of disease confirm pigeon intestinal mucin as an important antigen in PFL.


Molecular diagnostic testing for determination of myeloid lineage in acute leukemias.
Determination of myeloid vs. lymphoid cell lineage in acute leukemias is essential for diagnosis, prognosis, and treatment. However, some leukemic cells are too primitive to be identified based on conventional morphology, cytochemistry, and immunophenotype criteria. The gene for myeloperoxidase (MPO) is central to the lineage designation of myeloid cells and their function. Expression of MPO messenger ribonucleic acid (mRNA) may serve to determine the myeloid lineage of primitive leukemic cells that do not express the final product, MPO. Recently, a procedure has been developed for detection of MPO mRNA in leukemic cells, based on message amplification by reverse transcription-polymerase chain reaction (RT-PCR). Three variant polymerase chain reaction (PCR) methods, developed in view of future diagnostic applications of the RT-PCR procedure are reported: (A) nested PCR, using a second pair of internal primers for reamplification of the first PCR product; (B) multiplex PCR for concomitant amplification of up to seven sequences encompassing all regions of MPO mRNA; (C) adaptation of the standard PCR procedure and the two variant methods for use of cellular material from air-dried unstained or Wright stained smears. These variant procedures developed on the HL-60 cell line make the RT-PCR detection of MPO mRNA easily applicable as a new diagnostic test in acute leukemias and confer the versatility needed in the clinical setting. The advantages of our new procedure for MPO mRNA detection are: high sensitivity and specificity, speed, simplicity, and safety. Myeloid lineage of leukemic blasts can be established, confirmed, or ruled out in 6 to 10 hours, making results available at the time of bone marrow sign-out.(ABSTRACT TRUNCATED AT 250 WORDS)


Contributions of the oligopeptide permeases in multistep of Vibrio alginolyticus pathogenesis.
Vibrio alginolyticus has been associated with several diseases of cultivated marine animals, and has led to considerable economic losses. The oligopeptide permease (Opp) has been proven to play a variety of important roles in nutrition and virulence in several bacteria. In our previous research, the opp gene cluster was identified in Vibrio alginolyticus with transcriptome sequence, which also indicated that the Opp system might play roles in the regulation of adhesion. In this study, the relationship between V. alginolyticus virulence and the opp gene cluster was determined using gene silencing followed by RT-qPCR, in vitro adhesion assay, growth curves detection in the presence of glutathione (GSH) as a toxic substrate, hemolysis assay, biofilm assay, and artificial infection. Silencing these genes led to deficiencies in adhesion, peptide internalization, biofilm production, hemolytic activity, and virulence. The expression levels of hapr, hapa, tlh, and hlya, which are important genes closely related to the hemolytic activity of Vibrio, were significantly downregulated in all of the RNAi groups. Furthermore, the expression of oppA, oppB, oppC, oppD, and oppF was significantly influenced by temperature, starvation, and pH. These results indicate that (1) oppABCDF contributed in multistep of V. alginolyticus pathogenesis, including adhesion, biofilm production, and hemolytic activity; (2) oppABCDF was sensitive to different temperatures, changes in pH, and increased starvation time.


A Cross-Study Biomarker Signature of Human Bronchial Epithelial Cells Infected with Respiratory Syncytial Virus.
Respiratory syncytial virus (RSV) is a major cause of lower respiratory tract infections in children, elderly, and immunocompromised individuals. Despite of advances in diagnosis and treatment, biomarkers of RSV infection are still unclear. To understand the host response and propose signatures of RSV infection, previous studies evaluated the transcriptional profile of the human bronchial epithelial cell line-BEAS-2B-infected with different strains of this virus. However, the evolution of statistical methods and functional analysis together with the large amount of expression data provide opportunities to uncover novel biomarkers of inflammation and infections. In view of those facts publicly available microarray datasets from RSV-infected BEAS-2B cells were analyzed with linear model-based statistics and the platform for functional analysis InnateDB. The results from those analyses argue for the reevaluation of previously reported transcription patterns and biological pathways in BEAS-2B cell lines infected with RSV. Importantly, this study revealed a biosignature constituted by genes such as ABCC4, ARMC8, BCLAF1, EZH1, FAM118A, FAM208B, FUS, HSPH1, KAZN, MAP3K2, N6AMT1, PRMT2, S100PBP, SERPINA1, TLK2, ZNF322, and ZNF337 which should be considered in the development of new molecular diagnosis tools.


Activation of sirtuin 1/3 improves vascular hyporeactivity in severe hemorrhagic shock by alleviation of mitochondrial damage.
Vascular hyporeactivity is one of the major causes responsible for refractory hypotension and associated mortality in severe hemorrhagic shock. Mitochondrial permeability transition (mPT) pore opening in arteriolar smooth muscle cells (ASMCs) is involved in the pathogenesis of vascular hyporeactivity. However, the molecular mechanism underlying mitochondrial injury in ASMCs during hemorrhagic shock is not well understood. Here we produced an in vivo model of severe hemorrhagic shock in adult Wistar rats. We found that sirtuin (SIRT)1/3 protein levels and deacetylase activities were decreased in ASMCs following severe shock. Immunofluorescence staining confirmed reduced levels of SIRT1 in the nucleus and SIRT3 in the mitochondria, respectively. Acetylation of cyclophilin D (CyPD), a component of mPT pore, was increased. SIRT1 activators suppressed mPT pore opening and ameliorated mitochondrial injury in ASMCs after severe shock. Furthermore, administration of SIRT1 activators improved vasoreactivity in rats under severe shock. Our data suggest that epigenetic mechanisms, namely histone post-translational modifications, are involved in regulation of mPT by SIRT1/SIRT3- mediated deacetylation of CyPD. SIRT1/3 is a promising therapeutic target for the treatment of severe hemorrhagic shock.


ENaC- and CFTR-dependent ion and fluid transport in human middle ear epithelial cells.
Ion channels, such as the epithelial sodium channel (ENaC), are essential for maintaining a fluid-free middle ear cavity by controlling periciliary fluid. Deviations from the normal volume or compositions of periciliary fluid are probably responsible for otitis media with effusion. To elucidate the physiologic roles of the ENaC and cystic fibrosis transmembrane conductance regulator (CFTR) in the middle ear mucosa, we compared the electrophysiological activity and protein expressions of ENaC and CFTR in normal human middle ear epithelial (NHMEE) cells with those in normal human nasal epithelial (NHNE) cells. We also evaluated the role of ENaC and CFTR in fluid transport by NHMEE cells. Short-circuit current (Isc) was measured in cell monolayers by modified Ussing chambers. Immunoblotting was performed for ENaC and CFTR. In addition, transepithelial fluid transport was measured after loading 100 microl of fluid onto the luminal cell surface. The amiloride-sensitive Isc in NHMEE cells was much larger than in NHNE cells, whereas the forskolin-induced Isc, presumably mediated by CFTR, was significantly smaller in NHMEE cells. ENaC subunits alpha, beta, and gamma were all detected in NHMEE cells, and their expressions were stronger than those in NHNE cells. In comparison, CFTR was also detected in the middle ear mucosa, but at a lower expression level than in NHNE cells. NHMEE cells showed more amiloride-sensitive fluid absorption than NHNE cells. In contrast, fluid absorption was less sensitive to forskolin/IBMX in NHMEE cells than in NHNE cells. The ATP induced Cl- efflux and the amplitude of ATP-induced current in NHMEE cells was much larger than in NHNE cells. In the present study, we have demonstrated an enhanced amiloride-sensitive Isc and fluid absorption in NHMEE cells, where the role of CFTR is limited. Our data also suggest that the ATP-induced Cl- channel could be an alternative Cl- channel to CFTR in NHMEE cells.


[The relationship between within-day drinking occasion and intake amount of water of primary and middle school students in four cities of China].
To investigate the number of drinking occasions per day and average amount consumed per drinking occasion of primary and middle school students in four cities of China, and understand the relationship among drinking occasion, average amount consumed per drinking occasion and total drinking water. A total of 5914 primary and middle school students from Beijing, Shanghai, Guangzhou and Chengdu were selected using multiple-stage random sampling method, and 5868 students completed the study from September to October in 2011. The detailed information of amounts and types of daily drinking water was recorded by subjects using a 24 hours measurement for seven consecutive days. Analysis of the relationship among drinking occasion, average amount consumed per drinking occasion and total drinking water was carried out. The daily total drinking water of subjects was (1089 ± 540) ml; the daily number of drinking occasions was (4.7 ± 1.8) times, with 79.1% (4639/5868) of subjects reporting 6 or less drinking occasions. The amount consumed per drinking occasion was (239 ± 96) ml, plain water (231 ± 112) ml, and beverages (237 ± 112) ml. The number of drinking occasions of subjects was positively correlated with total drinking water (r = 0.614, P < 0.05), and negatively correlated with the average amount consumed per drinking occasion (r = -0.211, P < 0.05). Total drinking water and the average amount consumed per drinking occasion was positively correlated (r = 0.598, P < 0.05). The number of drinking occasion of primary and middle school students more than 6 times was fewer in four cities of China, but the average amount of beverages consumed per drinking occasion was relatively more. With the increasing of drinking occasion, the average amount consumed per drinking occasion decreased, but total drinking water increased.


Unilateral cortical application of interleukin-1beta (IL1beta) induces asymmetry in fos, IL1beta and nerve growth factor immunoreactivity: implications for sleep regulation.
Unilateral injection of interleukin-1 beta (IL1beta) into the somatosensory cortex enhances EEG slow wave activity ipsilaterally during non-rapid eye movement sleep [Yasuda, T., Yoshida, H., Garcia-Garcia, F., Kay, D., Krueger, J.M., 2005. Interleukin-1beta has a role in cerebral cortical state-dependent electroencephalographic slow-wave activity. Sleep 28, 177-184]. We show that a similar unilateral microinjection of IL1beta (10 ng) into layer VI or onto the surface of the primary somatosensory cortex induced increases in the neuronal activity marker, Fos, relative to the contralateral side that received saline or heat-inactivated IL1beta. When IL1beta was microinjected into layer VI, increases in Fos-immunoreactive nuclei were evident in layers II, III and VI of the somatosensory cortex and connected cortical regions, such as the endopiriform, secondary somatosensory, piriform and prefrontal cortex. Asymmetrical increases in Fos were also observed in subcortical regions, such as the reticular thalamus, which receives a main cortical projection, and hypothalamic regions implicated in sleep regulation, such as the ventrolateral preoptic area and dorsal median preoptic nucleus. Fos activation was not observed in many other brain regions. In the reticular thalamus and somatosensory cortex, the number of IL1beta-immunoreactive glial cells increased. Further, the number of NGF-immunoreactive cells in the primary somatosensory cortex and magnocellular preoptic nucleus increased on the IL1beta-injected side. These results are consistent with the hypothesis that sleep is initiated within the cortex after the local activation of specific cytokines and that whole organism sleep is coordinated via cortical connections with the subcortical sites.


2D/0D graphene hybrids for visible-blind flexible UV photodetectors.
Nitrogen-functionalized graphene quantum dots (NGQDs) are attractive building blocks for optoelectronic devices because of their exceptional tunable optical absorption and fluorescence properties. Here, we developed a high-performance flexible NGQD/graphene field-effect transistor (NGQD@GFET) hybrid ultraviolet (UV) photodetector, using dimethylamine-functionalized GQDs (NMe2-GQDs) with a large bandgap of ca. 3.3 eV. The NMe2-GQD@GFET photodetector exhibits high photoresponsivity and detectivity of ca. 1.5 × 104 A W-1 and ca. 5.5 × 1011 Jones, respectively, in the deep-UV region as short as 255 nm without application of a backgate voltage. The feasibility of these flexible UV photodetectors for practical application in flame alarms is also demonstrated.


Impaired expression and promotor hypermethylation of O6-methylguanine-DNA methyltransferase in retinoblastoma tissues.
To investigate the role of epigenetic changes in the promoter region of tumor-suppressor genes in the retinoblastoma genome and to study the disruption of expression of O6-methylguanine-DNA Methyltransferase (MGMT) due to aberrant methylation and its association with retinoblastoma. A series of 23 retinoblastoma tissue specimens and 2 retinoblastoma cell lines (Y79 and WERI-Rb1) were subjected to methylation-specific PCR (MSP) analysis of hypermethylated genes identified in human cancers, including p14(ARF), p15(INK4b), p16(INK4a), VHL, and MGMT. Further, the expression of MGMT was studied by immunohistochemistry and, when fresh tissue was available, by Western blot analysis and RT-PCR. Aberrant methylation of at least one MGMT locus was detected in 8 of the 23 tumors (35%), all of which (100%) had impaired or absent expression of MGMT. The remaining 15 tumor specimens were nonmethylated, and, among them, 7 (43%) showed defective expression. No methylation of tumor DNA was found on the p14(ARF), p15(INK4b), p16(INK4a), and VHL genes. Hypermethylation in the MGMT promoter was found to be prominently present in retinoblastoma with poor tissue differentiation, and was more frequently detected among patients with bilateral disease. Production of MGMT was consistent with expression of mRNA. No methylation of MGMT promoter was detected in the two retinoblastoma cell lines (Y79, WERI-Rb1). The data show a clear association between impaired production of MGMT and hypermethylation of the MGMT promoter, which appeared to relate to early onset and poor differentiation, suggesting that epigenetic silencing of MGMT by methylation of the promoter and reduced expression of MGMT may play an important role in the development and progression of retinoblastoma.


Organic Single-Crystal Semiconductor Films on a Millimeter Domain Scale.
Nucleation and growth processes can be effectively controlled in organic semiconductor films through a new concept of template-mediated molecular crystal seeds during the phase transition; the effective control of these processes ensures millimeter-scale crystal domains, as well as the performance of the resulting organic films with intrinsic hole mobility of 18 cm(2) V(-1) s(-1).


Perpendicular Iliac Screw Placement for Reinforcement of Spinopelvic Stabilization.
Iliac fixation is indicated to provide spinopelvic stabilization in select cases of long segment thoracolumbar spine fusion, spinal osteotomy/realignment, trauma, and instability caused by tumor or infection. Traditional iliac fixation with iliac screws or S2 alar/iliac (S2AI) screws may provide inadequate fixation in select clinical scenarios, such as severely compromised bone quality or spinopelvic dissociation. The purpose of this article is to describe the perpendicular iliac screw technique with ipsilateral iliac crest screw plus S2AI fixation. The technique may be applied for select individuals in cases of 3-column osteotomy in the lower lumbar spine, spinopelvic dissociation (caused by trauma, neoplasm, or Charcot arthropathy), and failure of previous iliac fixation and when anatomic constraints limit standard iliac screw or S2AI screw placement. [Orthopedics. 2016; 39(6):e1209-e1212.].


Giant true brachial artery aneurysm.
True brachial artery aneurysms (BAAs) are uncommon peripheral vessel aneurysms that typically occur in the setting of injury. While its relationship with trauma and infectious etiologies are well defined, the association between arteriovenous fistulas (AVFs) and BAA is less well understood. We present a case of a giant true BAA that presented several years post-AVF ligation. A review of BAA, its association with AVF, and its management follows.