[Cultivation of transgenic rice plants with OsCDPK7 gene and its salt tolerance].
A 1,700 bp DNA fragment, OsCDPK7 gene, was cloned with RT-PCR from liaoyan241 leaf treated under a low temperature of 4. Compared to the OsCDPK7 gene reported before (GenBank accession No. AB042550), this fragment, lack of 26 amino acids, possesses the activity of Ca2+-dependent protein kinase because of a complete integration of the Ca2+ binding structure domain and Ser/Thr protein kinase activity center. Plant expression vector was constructed,, OsCDPK7 gene was regulated by E12 promoter. OsCDPK7 gene was transferred into rice via Agrobacterium-mediated method. After Km screening and Southern blot, 10 transgenic plants were obtained. The analysis on the salt tolerance showed that the expression of OsCDPK7 gene composition enhanced the salt tolerance of T2 transgenic plants, part of T2 transgenic seeds could germinate in 0.2 mol/L NaCl medium, and T2 transgenic young plants could rejuvenate after treatment with 0.4 mol/L NaCl for 10 days, while the controlled plants could not germinate and died in salt stress. This research finding proved that the regulation factor of the plant signal transduction could enhance the salt tolerance of transgenic plants, while OsCDPK7 expression was different in the different tolerence transgenic plants.


Progesterone at high doses reduces the growth of U87 and A172 glioblastoma cells: Proteomic changes regarding metabolism and immunity.
While pregnancy may accelerate glioblastoma multiforme (GBM) growth, parity and progesterone (P4) containing treatments (ie, hormone replacement therapy) reduce the risk of GBM development. In parallel, low and high doses of P4 exert stimulating and inhibitory actions on GBM growth, respectively. The mechanisms behind the high-dose P4-suppression of GBM growth is unknown. In the present study, we assessed the changes in growth and proteomic profiles when high-dose P4 (100 and 300 µM) was administered in human U87 and A172 GBM cell lines. The xCELLigence system was used to examine cell growth when different concentrations of P4 (20, 50, 100, and 300 µM) was administered. The protein profiles were determined by two-dimensional gel electrophoresis in both cell lines when 100 and 300 µM P4 were administered. Finally, the pathways enriched by the differentially expressed proteins were assessed using bioinformatic tools. Increasing doses of P4 blocked the growth of both GBM cells. We identified 26 and 51 differentially expressed proteins (fc > 2) in A172 and U87 cell lines treated with P4, respectively. Only the pro-tumorigenic mitochondrial ornithine aminotransferase and anti-apoptotic mitochondrial 60 kDa heat shock protein were downregulated in A172 cell line and U87 cell line when treated with P4, respectively. Detoxification of reactive oxygen species, cellular response to stress, glucose metabolism, and immunity-related proteins were altered in P4-treated GBM cell lines. The paradox on the effect of low and high doses of P4 on GBM growth is gaining attention. The mechanism related to the high dose of P4 on GBM growth can be explained by the alterations in detoxification mechanisms, stress, and immune response and glucose metabolism. P4 suppresses GBM growth and as it is nontoxic in comparison to classical chemotherapeutics, it can be used as a new strategy in GBM treatment in the future.


A comparison of cochlear implant processing strategies in children using speech pattern audiometry.
The aim of this study was to use Speech Pattern Audiometry (SPA) to compare phoneme perception performance between SPEAK and ACE processing strategies with children using the Nucleus 24 cochlear implant. Listeners were evaluated in terms of ability to identify two synthetic word pairs, one signalling a voicing contrast and the other signalling a place of articulation contrast. For both word pairs, ACE was associated with more 'normal' phoneme boundaries than SPEAK. There was also a non-significant trend for better identification ability with ACE compared to SPEAK. The study suggests that there may be a place for SPA as part of the clinical battery of tests used for evaluation of cochlear implant speech processing strategies in children. Copyright (c) 2005 John Wiley & Sons, Ltd.


Spontaneous object-location memory based on environmental geometry is impaired by both hippocampal and dorsolateral striatal lesions.
We examined the role of the hippocampus and the dorsolateral striatum in the representation of environmental geometry using a spontaneous object recognition procedure. Rats were placed in a kite-shaped arena and allowed to explore two distinctive objects in each of the right-angled corners. In a different room, rats were then placed into a rectangular arena with two identical copies of one of the two objects from the exploration phase, one in each of the two adjacent right-angled corners that were separated by a long wall. Time spent exploring these two objects was recorded as a measure of recognition memory. Since both objects were in different locations with respect to the room (different between exploration and test phases) and the global geometry (also different between exploration and test phases), differential exploration of the objects must be a result of initial habituation to the object relative to its local geometric context. The results indicated an impairment in processing the local geometric features of the environment for both hippocampus and dorsolateral striatum lesioned rats compared with sham-operated controls, though a control experiment showed these rats were unimpaired in a standard object recognition task. The dorsolateral striatum has previously been implicated in egocentric route-learning, but the results indicate an unexpected role for the dorsolateral striatum in processing the spatial layout of the environment. The results provide the first evidence that lesions to the hippocampus and dorsolateral striatum impair spontaneous encoding of local environmental geometric features.


Mental health and risky sexual behaviors: evidence from DSM-IV Axis II disorders.
Several economic studies link poor mental health and substance misuse with risky sexual behaviors. However, none have examined the relationships between DSM-IV Axis II mental health disorders (A2s) and risky sexual behaviors. A2 disorders are a poorly understood, yet prevalent and disabling class of mental health conditions. They develop early in life through an interaction of genetics and environment, and are persistent across the life course. Common features include poor impulse control, addiction, social isolation, and elevated sexual desires, although the defining features vary substantially across disorder. To investigate the association between A2 disorders and three measures of risky sexual behavior. We obtain data on adults age 20 to 50 years from Wave II of the National Epidemiological Survey of Alcohol and Related Conditions (NESARC). Our outcome measures include early initiation into sexual activity, and past year regular use of alcohol before sex and sexually transmitted disease diagnosis. NESARC administrators use the Alcohol Use Disorder and Associated Disabilities Interview Schedule to classify respondents as meeting criteria for the ten A2 disorders recognized by the American Psychiatric Association. We construct several measures of A2 disorders based on the NESARC administrators' classifications. Given their comorbidity with A2 disorders, we explore the importance of Axis I disorders in the estimated associations. We find that A2 disorders are generally associated with an increase in the probability of risky sexual behaviors among both men and women. In specifications that disaggregate disorders into clusters and specific conditions, the significant associations are not uniform, but are broadly consistent with the defining features of the cluster or disorder. Inclusion of A1 disorders attenuates estimated associations for some risky sexual behaviors among men, but not for women. We find positive associations between A2 disorders and our measures of risky sexual behaviors. Our findings are subject to several data limitations, however. The NESARC lacks information on more advanced risky sexual behaviors and our measure of early initiation into sexual activity is retrospective. Identifying the causal effects of mental health and risky sexual behaviors is complicated due to bias from reverse causality and omitted variables. We believe these sources of bias are less of a concern in our study, however. Specifically, A2 disorders develop early in life and pre-date the risky sexual behaviors, thus negating reverse causality. Because the NESARC contains a rich set of personal characteristics, we are also able to minimize potential omitted variable bias. A2 disorders are significantly associated with risky sexual behaviors, which could lead to greater utilization and cost of health care services. Health care providers should consider A2 disorders when developing health promotion recommendations as these disorders may place individuals at elevated risk for unsafe sexual behaviors. Future studies should examine the causal mechanisms between A2 disorders and risky sexual behaviors.


Calsequestrin: more than 'only' a luminal Ca2+ buffer inside the sarcoplasmic reticulum.
In striated muscle, the sarcoplasmic reticulum (SR) Ca2+ release/ryanodine receptor (RyR) channel provides the pathway through which stored Ca2+ is released into the myoplasm during excitation-contraction coupling. Various luminal Ca2+-binding proteins are responsible for maintaining the free [Ca2+] at 10(-3)-10(-4) M in the SR lumen; in skeletal-muscle SR, it is mainly calsequestrin. Here we show that, depending on its phosphorylation state, calsequestrin selectively controls the RyR channel activity at 1 mM free luminal [Ca2+]. Calsequestrin exclusively in the dephosphorylated state enhanced the open probability by approx. 5-fold with a Hill coefficient (h) of 3.3, and increased the mean open time by about 2-fold, i.e. solely dephosphorylated calsequestrin regulates Ca2+ release from the SR. Because calsequestrin has been found to occur mainly in the phosphorylated state in the skeletal-muscle SR for the regulation of RyR channel activity, the dephosphorylation of calsequestrin would appear to be a quintessential physiological event.


Potassium and hypertension.
A recent meta-analysis supports the premise that increased potassium intake may play a role in the prevention and treatment of hypertension. The possible mechanisms by which potassium may act as an antihypertensive and recommendations regarding increasing potassium intake are also discussed.


An infectious clone of enterovirus 71(EV71) that is capable of infecting neonatal immune competent mice without adaptive mutations.
Enterovirus 71 (EV71) is a major pathogen that causes hand, foot and mouth disease (HFMD), which is a life threatening disease in certain children. The pathogenesis of EV71-caused HFMD is poorly defined due to the lack of simple and robust animal models with severe phenotypes that recapitulate symptoms observed in humans. Here, we generated the infectious clone of a clinical isolate from a severe HFMD patient. Virus rescued from the cDNA clone was infectious in cell lines. When administrated intraperitoneally to neonatal ICR, BALB/c and C57 immune competent mice at a dosage of1.4 × 104 pfu per mouse, the virus caused weight loss, paralysis and death in the infected mice after 4-5 days of infection. In the infected mice, detectable viral replication was detected in various tissues such as heart, liver, brain, lung, kidney, small intestine, leg skeletal muscle and medulla oblongata. The histology of the infected mice included massive myolysis, glomerular atrophy, villous blunting in small intestine, widened alveolar septum, diminished alveolar spaces and lymphocytes infiltration into the lung. By using the UV-inactivated virus as a control, we elucidated that the virus first amplified in the leg skeletal muscle tissue and the muscle tissue served as a primary viral replication site. In summary, we generated a stable EV71 infectious clone that is capable of infecting neonatal immune competent mice without adaptive mutations and provide a simple, valuable animal model for the studies of EV71pathogenesis and therapy.


Observational study of intravenous versus oral corticosteroids for acute asthma: an example of confounding by severity.
To demonstrate the problem of "confounding by severity" using the example of intravenous (IV) versus oral corticosteroids for the treatment of acute asthma. Double-blind, randomized trials have clearly demonstrated that IV and oral corticosteroids have comparable efficacy. Using a standardized protocol, 64 emergency departments enrolled 1,847 patients, aged 18-54 years, with acute asthma. Because route of corticosteroid therapy was not randomized, potential confounders of the association between corticosteroid route and hospital admission were controlled for by multivariate logistic regression and stratification. Among the 1,193 patients, 383 (32%) received IV corticosteroids and 810 (68%) received oral corticosteroids. The two groups differed markedly at baseline, with patients receiving IV corticosteroids having more severe asthma. Overall, patients receiving IV corticosteroids were more likely to be admitted or experience a relapse event within 48 hours (51% vs. 19%; p < 0.001). On multivariate analysis, patients receiving IV corticosteroids remained more likely to be admitted or experience a relapse event within 48 hours (odds ratio = 2.6; 95% confidence interval = 1.2 to 6.0). In this observational study, patients with worse asthma exacerbations were more likely to receive IV corticosteroids as compared with oral corticosteroids. Although we controlled for many markers of asthma severity, we were unable to completely control for baseline differences between the IV and oral corticosteroid groups. Observational research continues to serve as an important tool for describing problems and for understanding many exposure-disease associations. For examining the impact of treatments on adverse outcomes, randomized trials are often required to avoid intractable confounding by severity.


Design of an experimental setup for delivering intracortical microstimulation in vivo via Spiking Neural Network.
Electroceutical approaches for the treatment of neurological disorders, such as stroke, can take advantage of neuromorphic engineering, to develop devices able to achieve a seamless interaction with the neural system. This paper illustrates the development and test of a hardware-based Spiking Neural Network (SNN) to deliver neural-like stimulation patterns in an open-loop fashion. Neurons in the SNN have been designed by following the Hodgkin-Huxley formalism, with parameters taken from neuroscientific literature. We then built the set-up to deliver the SNN-driven stimulation in vivo. We used deeply anesthetized healthy rats to test the potential effect of the SNN-driven stimulation. We analyzed the neuronal firing activity pre- and post-stimulation in both the primary somatosensory and the rostral forelimb area. Our results showed that the SNN-based neurostimulation was able increase the spontaneous level of neuronal firing at both monitored locations, as found in the literature only for closed-loop stimulation. This study represents the first step towards translating the use of neuromorphic-based devices into clinical applications.Clinical Relevance- Stroke represents one of the leading causes of long-term disability and death worldwide. Intracortical microstimulation is an effective approach for restoring lost sensory motor integration by promoting plasticity among the affected brain areas. Stimulation delivered via neuromorphic-based open-loop systems (i.e. neuromorphic prostheses) can pave the way to novel electroceutical strategies for brain repair.


Decrease of tyrosine-O-sulfate-containing proteins found in rat fibroblasts infected with Rous sarcoma virus or Fujinami sarcoma virus.
Our interest was aroused by the recent report by Huttner [ Huttner , W. B. (1982) Nature (London) 299, 273-276] on general sulfation of tyrosine residues of proteins in normal and malignantly transformed tissues. Here we report on the reduction of sulfation in embryonic rat fibroblasts, line 3Y1, infected with Rous sarcoma virus or Fujinami sarcoma virus. In view of the instability of tyrosine O-sulfate in strong acid, the protein sulfation was tested for after incubation with [35S]sulfate and exhaustive Pronase hydrolysis. We found in general a reduction of sulfation in transformed tissue. It was greatest in the fibroblasts permanently transformed with Rous sarcoma virus. When fibroblasts transformed by the temperature-sensitive Fujinami sarcoma virus, line ts225 -3Y1, were used for comparison of sulfation at nonpermissive and permissive temperatures, the latter showed a strong reduction. Furthermore, we tested these cells for the uptake of inorganic [35S]sulfate. Uptake appeared highly reduced in the permanently infected fibroblasts, but ts225 -3Y1 grown at permissive and nonpermissive temperatures exhibited no difference. Uptake at both temperatures was comparable to uptake by normal 3Y1 cells. A recently much investigated cell surface protein, fibronectin, was reported to be lost on malignant transformation and to contain sulfate in an undetermined location. We found that ts225 -3Y1 cells grown at permissive temperature released fibronectin that contained tyrosine O-sulfate.


Mechanisms behind the stabilizing action of cellulose nanofibrils in wet-stable cellulose foams.
The principal purpose of the investigation was to clarify the mechanisms behind the stabilizing action of cellulose nanofibrils (CNFs) in wet-stable cellulose foams. Following the basic theories for particle-stabilized foams, the investigation was focused on how the surface energy of the stabilizing CNF particles, their aspect ratio and charge density, and the concentration of CNF particles at the air-water interface affect the foam stability and the mechanical properties of a particle-stabilized air-liquid interface. The foam stability was evaluated from how the foam height changed over time, and the mechanical properties of the interface were evaluated as the complex viscoelastic modulus of the interface using the pendant drop method. The most important results and conclusions are that CNFs can be used as stabilizing particles for aqueous foams already at a concentration as low as 5 g/L. The major reasons for this were the small dimensions of the CNF and their high aspect ratio, which is important for gel-formation and the complex viscoelastic modulus of the particle-filled air-water interface. The influence of the aspect ratio was also demonstrated by a much higher foam stability of foams stabilized with CNFs than of foams stabilized by cellulose nanocrystals (CNC) with the same chemical composition. The charge density of the CNFs affects the level of liberation within larger aggregates and hence also the number of contact points at the interface and the gel formation and complex viscoelastic modulus of the air-water interface. The charges also result in a disjoining pressure related to the long-range repulsive electrostatic pressure between particle-stabilized bubbles and hence contribute to foam stability.


Gene expression of antioxidant enzymes in experimental diabetic neuropathy.
Chronic hyperglycemia results in a large deficit in nerve blood flow. Both autoxidative- and ischemia-induced lipid peroxidation occurs, with resultant peripheral sensory neuropathy in streptozotocin-induced diabetes in the rat. Free radical defenses, especially involving antioxidant enzymes, have been suggested to be reduced, but scant information is available on chronic hyperglycemia. We evaluated the gene expression of glutathione peroxidase, catalase, and superoxide dismutase (cuprozinc and manganese separately) in L4,5 dorsal root ganglion (DRG) and superior cervical ganglion, as well as enzyme activity of glutathione peroxidase in DRG and sciatic nerve in experimental diabetic neuropathy of 3 months and 12 months durations. We also evaluated nerve electrophysiology of caudal, sciatic-tibial, and digital nerves. A nerve conduction deficit was seen in all nerves in experimental diabetic neuropathy at both 3 and 12 months. Gene expression of glutathione peroxidase, catalase, cuprozinc superoxide dismutase, and manganese superoxide dismutase were not reduced in experimental diabetic neuropathy at either 3 or 12 months. Catalase mRNA was significantly increased in experimental diabetic neuropathy at 12 months. Glutathione peroxidase enzyme activity was normal in sciatic nerve. We conclude that gene expression is not reduced in peripheral nerve tissues in very chronic experimental diabetic neuropathy. Changes in enzyme activity may be related to duration of diabetes or due to post-translational modifications.


Implementation of HIV pre-exposure prophylaxis for women of color: Perspectives from healthcare providers and staff from three clinical settings.
Women of color (WOC) account for 83% of new HIV infections among women in the United States. While pre-exposure prophylaxis (PrEP) is a safe, effective HIV prevention method for women, WOC are less likely to be prescribed PrEP than other populations. Guided by an implementation science research framework, we investigated the implementation of a PrEP initiative for WOC in a US city with high HIV incidence. Across three clinical sites, only three WOC were prescribed PrEP after one year. Analysis of qualitative interviews with clinic staff and providers identified time constraints, reluctance to prescribe PrEP, and discomfort with counseling as implementation barriers. Implementation facilitators included staff and leadership support for PrEP, alignment of PrEP services with organizational missions, and having a centralized PrEP Coordinator. By addressing these identified implementation barriers and facilitators, clinic staff and providers can ensure that WOC are provided with the full range of HIV prevention options.


Minimal incision total knee arthroplasty: early experience.
Total knee arthroplasty (TKA) has evolved during the past 30 years to a well-defined technical surgery with excellent results for as many as 20 years of followup. The concepts of exposure, ligament balancing, and joint alignment have been established clearly by Insall and others. In the early 1990s, Repicci introduced minimally invasive surgery for unicondylar knee arthroplasty. This approach suggested that less invasive surgery could accomplish similar results to those of standard unicondylar knee arthroplasty. Early reports in the literature support these conclusions. It only was logical that the minimally invasive surgery approach eventually would be applied to TKA. It is extremely important to establish a clear definition of the minimally invasive surgery and, then, to follow the results. We have completed 70 minimally-invasive TKAs during the past 9 months with early results that indicate less intraoperative blood loss, shorter length of stay, increased range of motion (ROM), with similar implant accuracy to standard TKA. These new procedures will require thorough evaluation as with any new clinical endeavor. It also is of paramount importance to remember that the main goal of any new technology is to advance the science of medicine without compromising the ultimate result for the patient. The early findings are encouraging for the future of minimally invasive TKA and we hope to improve the technology during the next few years.


The effective compounds and mechanisms of Cang-Fu-Dao-Tan Formula in treating polycystic ovary syndrome based on UPLC/Q-TOF-MS/MS, network pharmacology and molecular experiments.
Polycystic ovary syndrome (PCOS), as a common endocrine disease in reproductive-age women, which is characterized by both reproductive and metabolic disorders. Cang-Fu-Dao-Tan Formula (CFDTF) is an effective and relatively safe treatment for PCOS. However, the underlying mechanism is poorly understood. To explore the effective compounds and mechanisms of CFDTF in treating PCOS based on UPLC/Q-TOF-MS/MS, network pharmacology and molecular experiments. The UPLC/Q-TOF-MS/MS and TCMSP, SwissTargetPrediction databases were used to identify the active ingredients of CFDTF. Then GeneCards, Disgenet, Drugbank databases were used to obtain the PCOS related targets. Based above, the Drug-component-target (D-C-T) network and protein-protein-interaction (PPI) network were built to analysis the key targets. The Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis were performed to find the potential mechanisms. Finally, molecular docking analysis, molecular dynamics (MD) simulations and molecular experiments were used to confirm the interactions among the active compounds, targets and explore the potential mechanisms. A total of 20 compounds were identified by UPLC/Q-TOF-MS/MS, and 136 active compounds by TCMSP from CFDTF. After removing the duplicate results, there were 370 targets related to both CFDTF and PCOS, among which, MAPK3, AKT1, RELA, EGF, TP53 and MYC were proved to have high interactions with the components. The mechanisms of CFDTF against PCOS were related to PI3K-Akt, mTOR, MAPK signaling pathways, and the in vitro experiments proved that the CFDTF positively regulated the cell proliferation and inhibited the apoptosis levels in PCOS cell model. The combination of UPLC/Q-TOF-MS/MS, systematic network pharmacology and molecular experiments identified that the quercetin, hesperidin, and glycyrrhizin disaccharide are the TOP 3 effective compounds of CFDTF in treating PCOS and the potential mechanisms may involve in regulating proliferation and apoptosis of granulosa cells.


[Swelling of neurosecretory fibers (Herring bodies) in the reorganized hypophyseal stalk following hypophysectomy].
Herring bodies were revealed in the reorganized hypophyseal stalk of rats two months after hypophysectomy both in normal laboratory condition and under salt load. We observed Herring bodies with a storage of neurosecretory granules and neurohormones, as well as with massive destruction and disappearance of neurosecretory granules due to release of hormones in neuroplasm. Many Herring bodies were characterized by degenerative changes probably associated with aging of neurosecretory cells or their lesion due to the operation. Occasionally we revealed Herring bodies which contained a network of dilated neurotubules, that probably reflected either the reparative phase of secretory cycle of corresponding neurosecretory cells or restoration of their functions after hypophysectomy. Herring bodies were often situated around capillaries, but they were separated from perivascular space with narrow sprouts of pituicytes.


Effect of distraction osteogenesis of the mandible on upper airway volume and resistance in children with micrognathia.
Children with craniofacial anomalies often have compromise of the upper airway, a condition with potential for morbidity and mortality. In children with microretrognathia, the diminutive size and retruded position of the mandible reduces the size of the oropharynx, thereby predisposing to glossoptosis and airway obstruction. Although several authors have reported successful use of mandibular distraction osteogenesis to alleviate this type of upper airway obstruction, the physiologic relationship between changes in mandibular shape, size, and position and upper airway dynamics remains undefined. The purpose of this study was to develop methodologies to quantitatively evaluate upper airway dynamics in children with micrognathia both before and after mandibular distraction osteogenesis. The patient population consisted of four children with micrognathia who had successfully undergone upper airway stabilization by bilateral mandibular distraction osteogenesis. The data used were digitally archived computed tomographic scan data from high-resolution, thin-slice head computed tomographic scans obtained before and after mandibular distraction. Upper airway evaluation was performed in two ways: static and dynamic. Static analysis consisted of computer quantification of predistraction and postdistraction mandibular and upper airway volumes using Analyze imaging software. Dynamic analysis consisted of fabrication of rigid stereolithographic hollow cast models of the upper airway produced from computed tomographic scan data. Models were used for characterization of upper airway resistance and flow patterns as related to respiration. After distraction osteogenesis, mandibular total volume increased 32, 32, 18, and 25 percent (mean, 27 percent) and upper airway volume increased by 20, 31, 23, and 71 percent (mean, 37 percent). A significant decrease in flow resistance, both inspiratory and expiratory, was observed in the patient with the greatest upper airway volume increase (71 percent) after distraction. After distraction, the inspiratory resistance was diminished by 51 percent and the expiratory resistance diminished by 85 percent. However, the three patients with more modest upper airway volume increases of 20 to 31 percent demonstrated no statistically significant change in flow resistance after distraction. Results of this study support the conclusion that distraction osteogenesis of the micrognathic mandible increases the volume of the upper airway, roughly paralleling the increase in mandibular volume. In the biomechanical airway model studied, upper airway volume expansion has been shown to be able to decrease the flow resistance over the length of the airway, presumably secondary to an increase in the average cross-sectional area. The artificial rigidity of the stereolithographic "airway" compared with the elasticity of the human upper airway may account for the insensitivity of this model to smaller but clinically significant airway changes.


Histidine-stimulated divalent metal uptake in human erythrocytes and in the erythroleukaemic cell line HEL.92.1.7.
The uptake of 65Zn by human erythrocytes was investigated in the presence of high (40 mM) and low (5 mM) concentrations of histidine and 0-500 microM cobalt, nickel, manganese and zinc. Varying concentrations of metal mono- and bis-histidine complexes will be formed and the inhibition of 65Zn uptake could be correlated with the calculated complex concentrations to investigate competition between metals. For each metal, the calculated concentrations of bis-histidine complex giving 50% inhibition of 65Zn uptake were similar at both 5 mM and 40 mM histidine. Manganese-bis-histidine appeared to have a much higher affinity for the binding site than the other metal-bis-histidine complexes, which had similar affinities to each other. Studies of the inhibition of histidine-stimulated 54Mn uptake by the addition of manganese confirmed that manganese-bis-histidine does act as a substrate for the transporter in a similar fashion to the other metals studied. In addition, human erythroleukaemic cells (HEL cells) were used as a model for erythroid precursor cells. L-histidine, but not D-histidine, stimulated 65Zn uptake in a saturable fashion. The other metals competed with zinc in a similar manner to that seen in erythrocytes, and the affinity for manganese-bis-histidine was much greater than for the bis-histidine complexes of the other three metals. Both the capacity for metal transport per cell, and the affinity of the transporter for the metal-bis-histidine complexes, were much greater in the HEL cells than in the erythrocyte. It is suggested that histidine-stimulated metal transport may play a role in the supply of metals to maturing erythroid cells.


Jejunal perforation caused by a feeding jejunostomy tube: a case report.
Percutaneous endoscopic gastrostomy and feeding jejunostomy are used for providing long-term nutritional support to patients with neurological disorders. Various mechanical complications of these procedures are described. We report a case of a 17-year-old boy with cerebral injury who had a percutaneous endoscopic gastrostomy tube changed to a feeding jejunostomy tube. Twenty-four hours later he developed abdominal pain and became clinically septic. A contrast study through the feeding tube and a subsequent computed tomography scan did not reveal any intra-abdominal pathology. At laparotomy it was discovered that the tip of the feeding tube had perforated through the jejunal wall and was lying outside the lumen. This was successfully treated by re-inserting a feeding jejunostomy tube distally and closure of the perforation and previous FJ site We suggest that the threshold for contrast studies and operative intervention should be low in neurologically impaired patients to avoid the delay in treatment of tube-related complications.


Spatial distribution of triatomines (Reduviidae: Triatominae) in urban areas of the city of Salvador, Bahia, Brazil.
Environmental changes have a strong influence on the emergence and/or reemergence of infectious diseases. The city of Salvador, Brazil--currently the focus of a housing boom linked to massive deforestation--is an example in point as the destruction of the remaining areas of the Atlantic Forest around the city has led to an increased risk for Chagas disease. Human domiciles have been invaded by the triatomine vectors of Trypansoma cruzi, the flagellate protozoan causing Chagas disease, a problem of particular concern in urban/suburban areas of the city such as the Patamares sector in the north-east, where numbers of both the vector and human cases of the disease have increased lately. To control and prevent further deterioration of the situation, the control programme for Chagas disease, developed by the Bahia Center for Zoonosis Control, has divided the area into a grid of designated surveillance units (ZIs) that are subjected to vector examination. In six out of 98 of these ZIs, 988 triatomes were collected and georeferenced during the 3-year period between 2006 and 2009. The hottest months, that are also generally the driest, showed the highest numbers of triatomines with Triatoma tibiamaculata being the predominant species (98.3%) with Panstrongylus geniculatus present only occasionally (0.6%). Fifty-four percent of all triatomines captured were found inside the homes, and 48.6% out of 479 individuals in the affected ZIs selected for analysis tested positive for T. cruzi infection. The study presented here is a pioneering initiative to map the spatial distribution of triatomines based on geographical information systems with the additional aim of contributing to an expanded knowledge-base about T. cruzi and its vectors in urban areas and raise public health awareness of the risks involved.


Characterization of Volatile Compounds by SPME-GC-MS During the Ripening of Kedong Sufu, a Typical Chinese Traditional Bacteria-Fermented Soybean Product.
The purpose of the present study was to evaluate the volatile profile of Kedong sufu, which is a typical bacteria-fermented soybean product in China, using solid phase microextraction coupled to gas chromatography and mass spectrometry and to reveal the evolution and diversity of flavor substances for this specialty. A total of 75 compounds were identified, including 35 esters, 4 alcohols, 4 phenols, 4 aldehydes, 7 acids, 10 ketones, and 11 other compounds from sufu samples during ripening. Some volatile compounds increased with ripening time, especially hexadecenoic acid ethyl ester, methoxy acetic acid pentyl ester, benzene propanoic acid ethyl ester, ethyl 9-hexadecenoate, ethyl oleate, ethanol, 3-methyl-1-butanol, 5-methoxy-1-pentanol, and eugenol; these compounds enriched the flavors and provided the typical savory taste of Kedong sufu. PRACTICAL APPLICATION: This research elucidated the formation of flavor substances in sufu. For traditional fermented foods, this study provides a scientific basis for promoting the generation of typical flavor substances and for the precise determination of maturity time.


The effects of physical activity on psychogenic movement disorders.
Psychogenic movement disorders (PMD) are a diagnostic fascinating challenge in both neurologic and psychiatric setting. Many factors influence response to treatment, but few treatment strategies are available. Physical activity proves to be effective in the treatment of depression, anxiety, and other psychiatric disorders, but the effects of regular walking exercise on patients with PMD have never been investigated in a single-blind study. Sixteen outpatients [13 women; mean age 33.0 years (range 22-51)] with primarily mild-to-moderate PMD completed a thrice-weekly, 12-weeks mild walking program. Assessments included DSM-IV interview, the Psychogenic Movement Disorder Rating Scale (PMDRS), Beck Anxiety Inventory (BAI), Hamilton Depression Scale (HDS), V02 Max, and body mass index (BMI). Changes in total score on the PMDRS were the primary endpoint. A comparison of all measures taken at study onset and after completing the exercise program indicates statistically significant improvements. We observed a relevant improvement in 10 of 16 patients (62%). The mean difference for the primary outcome (PMDRS total) corresponded to about 70%. Compliance was good, and there were no adverse effects. This study provides preliminary evidence for regular low-medium intensity exercise as a safe, adequate, and pleasing intervention for PMD. Furthermore, well-designed studies appear justified to confirm these findings.


Mutagenicity and genotoxicity of PM2.5 issued from an urbano-industrialized area of Dunkerque (France).
Epidemiological studies have demonstrated the link between chronic exposure to particulate matter (PM), especially particles with an aerodynamic diameter lesser than 2.5 µm (PM(2.5) ), and lung cancer. Mechanistic investigations focus on the contribution of the various genotoxicants adsorbed onto the particles, and more particularly on polycyclic aromatic hydrocarbons or nitroaromatics. Most of the previous studies dealing with genotoxic and/or mutagenic measurements were performed on organic extracts obtained from PM(2.5) collected in polluted areas. In contrast, we have evaluated genotoxic and mutagenic properties of urbano-industrial PM(2.5) (PM) collected in Dunkerque (France). Thermally desorbed PM(2.5) (dPM) was also comparatively studied. Suspensions of PM and dPM (5-50 µg per plate) were tested in Salmonella tester strains TA98, TA102 and YG1041 ± S9mix. Significant mutagenicity was observed for PM in YG1041 ± S9 mix. In strain TA102 - S9mix, a slight, but not significant dose-response increase was observed, for both PM and dPM. Genotoxic properties of PM and dPM were evaluated by the measurement of (1) 8-OHdG in A549 cells and (2) bulky DNA adducts on A549 cells and on human alveolar macrophages (AMs) in primary culture. A dose-dependant formation of 8-OHdG adducts was observed on A549 cells for PM and dPM, probably mainly attributed to the core of the particles. Bulky DNA adducts were observed only in AMs after exposure to PM and dPM. In conclusion, using relevant exposure models, suspension of PM(2.5) induces a combination of DNA-interaction mechanisms, which could contribute to the induction of lung cancer in exposed populations.


Intervention of Bro1 in pH-responsive Rim20 localization in Saccharomyces cerevisiae.
Yeast cells contain two Bro1 domain proteins: Bro1, which is required for endosomal trafficking, and Rim20, which is required for the response to the external pH via the Rim101 pathway. Rim20 associates with endosomal structures under alkaline growth conditions, when it promotes activation of Rim101 through proteolytic cleavage. We report here that the pH-dependent localization of Rim20 is contingent on the amount of Bro1 in the cell. Cells that lack Bro1 have increased endosomal Rim20-green fluorescent protein (GFP) under acidic conditions; cells that overexpress Bro1 have reduced endosomal Rim20-GFP under acidic or alkaline conditions. The novel endosomal association of Rim20-GFP in the absence of Bro1 requires ESCRT components including Vps27 but not specific Rim101 pathway components such as Dfg16. Vps27 influences the localization of Bro1 but is not required for RIM101 pathway activation in wild-type cells, thus suggesting that Rim20 enters the Bro1 localization pathway when a vacancy exists. Despite altered localization of Rim20, the lack of Bro1 does not bypass the need for signaling protein Dfg16 to activate Rim101, as evidenced by the expression levels of the Rim101 target genes RIM8 and SMP1. Therefore, endosomal association of Rim20 is not sufficient to promote Rim101 activation.


Effects of a grapeseed procyanidin extract (GSPE) on insulin resistance.
Flavonoids are beneficial compounds against risk factors for metabolic syndrome, but their effects and the mechanisms on glucose homeostasis modulation are not well defined. In the present study, we first checked the efficacy of grapeseed procyanidin extract (GSPE) for stimulating glucose uptake in insulin-resistant 3T3-L1 adipocytes. Results show that when resistance is induced with chronic insulin treatment, GSPE maintain a higher stimulating capacity than insulin. In contrast, when dexamethasone is used as the resistance-inducing agent, GSPE is less effective. Next we evaluated how effective different GSPE treatments are at improving glucose metabolism in hyperinsulinemic animals (fed a cafeteria diet). GSPE reduced plasma insulin levels. The lower dose (25 mg GSPE/kg body weight per day) administered for 30 days improved the HOmeostasis Model Assessment-insulin resistance index. This was accompanied by down-regulation of Pparg2, Glut4 and Irs1 in mesenteric white adipose tissue. Similarly, a chronic GSPE treatment of insulin-resistant 3T3-L1 adipocytes down-regulated the mRNA levels of those adipocyte markers, although cells were still able to respond to the acute stimulation of glucose uptake. In summary, 25 mg/kg body weight per day of GSPE has a positive long-term effect on glucose homeostasis, and GSPE could be targeted at adipose tissue, where it might directly stimulate glucose uptake. This work also highlights the need to carefully consider the bioactive dose, since a higher dose does not necessarily correlate to a greater positive effect.


Selective metabolic effects of gold nanorods on normal and cancer cells and their application in anticancer drug screening.
Cetyltrimethylammonim bromide coated gold nanorods (Au NRs) has a potential to become anti-cancer nano-drugs. Previously, the comparative responses of human alveolar adenocarcinoma epithelial cells (A549) and normal bronchial epithelial cells (16HBE) exposed to Au NRs have been characterized. It has been shown that Au NRs are translocated from the lysosome to the mitochondria in A549 cells but not in normal 16HBE cell lines. However, the molecular information during this cellular translocation remains largely undetermined. Here, we have used a metabonomic technique to comparatively analyze the time-dependent metabolic changes in Au NRs-induced A549 and 16HBE. We found that Au NRs exposure caused a disruption in the intracellular environment of both A549 and 16HBE cells, which metabolically manifested in the reduction of lactate levels in both cell lines. In addition, Au NRs induced oxidative stress in both cells lines. However, the 16HBE cells are more able to offset the oxidative stress than the A549 cells; this is because de novo GSH synthesis is triggered in Au NRs treated 16HBE cells but not in A549 cells, and the conversion of GSH to GSSG is more profound in 16HBE cells compared to A549 cells. The severe oxidative stress induces damage to mitochondria in A549 cells, leading to cell death, which is evident in the marked reduction in the levels of nucleosides and nucleotides. Furthermore, significantly elevated levels of amino acids are likely due to stress hormones being produced in Au NRs treated cells. These findings provide comprehensive molecular information on the distinctive intracellular localization, cellular uptake and translocation of Au NRs in normal and tumor cells, highlighting the value of metabonomics in assessing biological effects of nano-drugs.


Designing highly emissive over-1000 nm near-infrared fluorescent dye-loaded polystyrene-based nanoparticles for in vivo deep imaging.
Polystyrene-based nanoparticles (PSt NPs) prepared by emulsion polymerization are promising organic matrices for encapsulating over-thousand-nanometer near-infrared (OTN-NIR) fluorescent dyes, such as thiopyrilium IR-1061, for OTN-NIR dynamic live imaging. Herein, we propose an effective approach to obtain highly emissive OTN-NIR fluorescent PSt NPs (OTN-PSt NPs) in which the polarity of the PSt NPs was adjusted by changing the monomer ratio (styrene to acrylic acid) in the PSt NPs and the dimethyl sulfoxide concentration in the IR-1061 loading process. Moreover, OTN-PSt NPs covalently modified with poly(ethylene glycol) (PEG) (OTN-PSt-PEG NPs) showed high dispersion stability under physiological conditions and minimal cytotoxicity. Notably, the optimized OTN-PSt-PEG NPs were effective in the dynamic live imaging of mice. This methodology is expected to facilitate the design of certain polar thiopyrilium dye-loaded OTN-NIR fluorescent imaging probes with high emissivity.


Double-stranded gap repair of DNA by gene conversion in Escherichia coli.
We demonstrated repair of a double-stranded DNA gap through gene conversion by a homologous DNA sequence in Escherichia coli. We made a double-stranded gap in one of the two regions of homology in an inverted orientation on a plasmid DNA molecule and introduced it into an E. coli strain which has the RecE system of recombination (genotype; sbcA23 recB21 recC22). We detected repair products by genetic selection. The repair products were those expected by the double-strand-gap repair model. Gene conversion was frequently accompanied by crossing over of the flanking sequences as in eukaryotes. This double-strand gap repair mechanism can explain plasmid recombination in the absence of an artificial double-stranded break reported in a companion study by Yamamoto et al.


Paradigms of Dynamic Control of Thyroid Hormone Signaling.
Thyroid hormone (TH) molecules enter cells via membrane transporters and, depending on the cell type, can be activated (i.e., T4 to T3 conversion) or inactivated (i.e., T3 to 3,3'-diiodo-l-thyronine or T4 to reverse T3 conversion). These reactions are catalyzed by the deiodinases. The biologically active hormone, T3, eventually binds to intracellular TH receptors (TRs), TRα and TRβ, and initiate TH signaling, that is, regulation of target genes and other metabolic pathways. At least three families of transmembrane transporters, MCT, OATP, and LAT, facilitate the entry of TH into cells, which follow the gradient of free hormone between the extracellular fluid and the cytoplasm. Inactivation or marked downregulation of TH transporters can dampen TH signaling. At the same time, dynamic modifications in the expression or activity of TRs and transcriptional coregulators can affect positively or negatively the intensity of TH signaling. However, the deiodinases are the element that provides greatest amplitude in dynamic control of TH signaling. Cells that express the activating deiodinase DIO2 can rapidly enhance TH signaling due to intracellular buildup of T3. In contrast, TH signaling is dampened in cells that express the inactivating deiodinase DIO3. This explains how THs can regulate pathways in development, metabolism, and growth, despite rather stable levels in the circulation. As a consequence, TH signaling is unique for each cell (tissue or organ), depending on circulating TH levels and on the exclusive blend of transporters, deiodinases, and TRs present in each cell. In this review we explore the key mechanisms underlying customization of TH signaling during development, in health and in disease states.


The Na+/Ca2+ exchange blocker SEA0400 fails to enhance cytosolic Ca2+ transient and contractility in canine ventricular cardiomyocytes.
This study was designed to evaluate the effects of the Na(+)/Ca(2+) exchange (NCX) inhibitor SEA0400 on Ca(2+) handling in isolated canine ventricular myocytes. Intracellular Ca(2+) ([Ca(2+)](i)) transients, induced by either field stimulation or caffeine flush, were monitored using Ca(2+) indicator dyes. [Ca(2+)](i)-dependent modulation of the inhibitory effect of SEA0400 on NCX was characterized by the changes in Ni(2+)-sensitive current in voltage-clamped myocytes. Sarcoplasmic reticulum (SR) Ca(2+) release and uptake were studied in SR membrane vesicles. Gating properties of single-ryanodine receptors were analysed in lipid bilayers. Ca(2+) sensitivity of the contractile machinery was evaluated in chemically skinned myocytes. In myocytes paced at 1 Hz, neither diastolic [Ca(2+)](i) nor the amplitude of [Ca(2+)](i) transients was significantly altered by SEA0400 up to the concentration of 1 microM, which was shown to inhibit the exchange current. The blocking effect of SEA0400 on NCX decreased with increasing [Ca(2+)](i), and it was more pronounced in reverse than in forward mode operation at every [Ca(2+)](i) examined. The rate of decay of the caffeine-induced [Ca(2+)](i) transients was decreased significantly by 1 microM SEA0400; however, this effect was only a fraction of that observed with 10 mM NiCl(2). Neither SR Ca(2+) release and uptake nor cell shortening and Ca(2+) sensitivity of the contractile proteins were influenced by SEA0400. The lack of any major SEA0400-induced shift in Ca(2+) transients or contractility of myocytes can well be explained by its limited inhibitory effect on NCX (further attenuated by elevated [Ca(2+)](i) levels) and a concomitant reduction in Ca(2+) influx due to the predominantly reverse mode blockade of NCX and suppression of L-type Ca(2+) current.


Consumption of Boiled, but Not Grilled, Roasted, or Barbecued Beef Modifies the Urinary Metabolite Profiles in Rats.
The consumption of processed meat is associated with increased risk of chronic diseases, but determining how the exposure to specific cooking processes alters the metabolome is an analytical challenge. This study aims to evaluate the impact of four typical cooking methods for beef (boiling, barbecuing, grilling, and roasting) on the urinary metabolite profiles in rats, using a non-targeted approach. Male Wistar rats (n = 48) are fed for 3 weeks with experimental diets containing either raw or cooked (boiled, barbecued, grilled, and roasted) beef. A control group is fed with milk proteins. The 24 h-urines are analyzed using LC-MS. The consumption of boiled meat leads to the specific excretion of di- and tri-peptides (aspartyl-leucine, glycyl-aspartate, and aspartyl-prolyl-threonine) and a cyclo-prolyl-proline (p < 0.001). No singular metabolite specifically associated with the groups "grilled," "roasted," and "barbecued" meat is observed. Urinary metabolite profiles of rats fed boiled beef are clearly distinct from those of rats fed with raw, grilled, roasted, or barbecued beef. The specific metabolites include the products of non-digested proteins and may be useful as potential intake biomarkers of this meat cooking method.


Bacteriologic cultural examination of the lower respiratory tract of laboratory dogs.
During lung transplantation experiments, the lower respiratory tracts of dogs were examined by bacteriologic cultural technique pre- and postoperatively. Three methods were used to obtain culture material: transtracheal aspiration, lung puncture, and lung biopsy collection. Both before and after operation, infections were commonplace. Antibiotics given at the time of operation tended to suppress infection.


Functional analysis of disease-causing mutations in human galactokinase.
Galactokinase (EC 18.104.22.168) catalyzes the first committed step in the catabolism of galactose. The sugar is phosphorylated at position 1 at the expense of ATP. Lack of fully functional galactokinase is one cause of the inherited disease galactosemia, the main clinical manifestation of which is early onset cataracts. Human galactokinase (GALK1) was expressed in and purified from Escherichia coli. The recombinant enzyme was both soluble and active. Product inhibition studies showed that the most likely kinetic mechanism of the enzyme was an ordered ternary complex one in which ATP is the first substrate to bind. The lack of a solvent kinetic isotope effect suggests that proton transfer is unlikely to be involved in the rate determining step of catalysis. Ten mutations that are known to cause galactosemia were constructed and expressed in E. coli. Of these, five (P28T, V32M, G36R, T288M and A384P) were insoluble following induction and could not be studied further. Four of the remainder (H44Y, R68C, G346S and G349S) were all less active than the wild-type enzyme. One mutant (A198V) had kinetic properties that were essentially wild-type. These results are discussed both in terms of galactokinase structure-function relationships and how these functional changes may relate to the causes of galactosemia.


Astatotilapia burtoni: A Model System for Analyzing the Neurobiology of Behavior.
Most biomedical research is performed using a very limited number of "model" species. In part, this has resulted from a combination of full genomes, manipulation of genes, and short generation times in these species. However, the advent of low-cost sequencing and gene editing in any organism has increased the use of nontraditional organisms. Many scientists have paraphrased the adage by Krogh [ Krogh , A. ( 2018 ) Science 70 , 200 - 204 ] that for many biological problems some species will prove to be most convenient and useful to study. In particular, using organisms most suited to the specific research question can lead to novel insights about fundamental physiological, neurobiological, immunological, and neuroendocrine systems that can advance our understanding of the well-being and health of humans. In addition, such studies have led to new ideas about the evolution and mechanisms that control social behavior. Fishes constitute about 50% of all vertebrate species and are the most diverse vertebrate radiation. Here we review behavioral and neurobiological discoveries of plasticity in social behavior resulting from analysis of an African cichlid fish, showing how its unique behavioral system has facilitated a broad range of discoveries. For many future questions, Astatotilapia burtoni and other cichlid fishes may be ideally suited to study as we advance our understanding of the neural basis of social decisions.


Contribution of autoallergy to the pathogenesis in the NOD mice.
The immunoglobulin isotype IgE is commonly associated with allergy. However, its involvement in autoimmune disease in general, and Type 1 diabetes (T1D) in particular, is still not completely clarified, nonetheless IgE has been observed in patients with T1D. In this article, we aimed to elucidate the contribution of IgE in the pathogenesis of the disease in a spontaneous model for T1D, i.e. the NOD mouse. We observed increased levels of IgE in splenic, lymph node and peripheral blood B cells in the NOD mice compared to the control C57BL/6 (B6) mice. No correlation was found between the IgE levels on B cells and those in the sera of these mice, indicating a B cell intrinsic property mediating IgE capture in NOD. Functionally, the B cells from NOD were similar to B6 in rescuing the IgE-mediated immune response via the low affinity receptor CD23 in a transgenic adoptive transfer system. However, the involvement of IgE in diabetes development was clearly demonstrated, as treatment with anti-IgE antibodies delayed the incidence of the diabetes in the NOD mice compared to the PBS treated group. Pancreas sections from a 13-week-old NOD revealed the presence of tertiary lymphoid structures with T cells, B cells, germinal centers and IgE suggesting the presence of autoantigen specific IgE. Our study provides an insight to the commonly overlooked immunoglobulin IgE and its potential role in autoimmunity.


Diet and apoptosis.
A range of compounds in or derived from the diet modulates apoptosis in cell cultures in vitro. These observations have important implications concerning the mechanisms whereby dietary components affect health. Proapoptotic compounds could protect against cancer by enhancing elimination of initiated, precancerous cells, and antiapoptotic compounds could promote tumor formation by inhibiting apoptosis in genetically damaged cells. Proapoptotic compounds could also contribute to age-related degenerative diseases by activating cell death in postmitotic cells or shifting the normal balance of mitosis and apoptosis in tissues with regenerative capacity. Many age-related diseases, for example macular degeneration and Parkinson's disease, appear to have oxidative stress as an underlying component that interacts with genetic, dietary, and environmental factors to determine relative risk in an individual. Oxidative stress activates apoptosis, and antioxidants protect against apoptosis in vitro; thus, a central role of dietary antioxidants may be to protect against apoptosis. However, little in vivo data are available to directly link diet with altered apoptosis as an underlying determinant of disease. Moreover, the possible antagonistic effects of different dietary components and the uncertainty about whether proapoptotic compounds that may protect against cancer could contribute to degenerative diseases and vice versa indicate that there is a great need for better in vivo assessment of apoptosis and that caution should be exercised when extrapolating in vitro data on apoptosis to in vivo dietary recommendations.


Dual Requirement of Cytokine and Activation Receptor Triggering for Cytotoxic Control of Murine Cytomegalovirus by NK Cells.
Natural killer (NK) cells play a critical role in controlling murine cytomegalovirus (MCMV) and can mediate both cytokine production and direct cytotoxicity. The NK cell activation receptor, Ly49H, is responsible for genetic resistance to MCMV in C57BL/6 mice. Recognition of the viral m157 protein by Ly49H is sufficient for effective control of MCMV infection. Additionally, during the host response to infection, distinct immune and non-immune cells elaborate a variety of pleiotropic cytokines which have the potential to impact viral pathogenesis, NK cells, and other immune functions, both directly and indirectly. While the effects of various immune deficiencies have been examined for general antiviral phenotypes, their direct effects on Ly49H-dependent MCMV control are poorly understood. To specifically interrogate Ly49H-dependent functions, herein we employed an in vivo viral competition approach to show Ly49H-dependent MCMV control is specifically mediated through cytotoxicity but not IFNγ production. Whereas m157 induced Ly49H-dependent degranulation, efficient cytotoxicity also required either IL-12 or type I interferon (IFN-I) which acted directly on NK cells to produce granzyme B. These studies demonstrate that both of these distinct NK cell-intrinsic mechanisms are integrated for optimal viral control by NK cells.


Surgical criteria for correcting the overlapping fifth toe.
Overlapping fifth toe, which is a varus deformity, usually occurs bilaterally and is a common familial deformity. It may be mild or severe and symptomatic or asymptomatic. The author discusses eight patients with overlapping varus deformity of the fifth toe and the various surgical procedures that may be used for correction.


Reproducibility of R-fMRI metrics on the impact of different strategies for multiple comparison correction and sample sizes.
Concerns regarding reproducibility of resting-state functional magnetic resonance imaging (R-fMRI) findings have been raised. Little is known about how to operationally define R-fMRI reproducibility and to what extent it is affected by multiple comparison correction strategies and sample size. We comprehensively assessed two aspects of reproducibility, test-retest reliability and replicability, on widely used R-fMRI metrics in both between-subject contrasts of sex differences and within-subject comparisons of eyes-open and eyes-closed (EOEC) conditions. We noted permutation test with Threshold-Free Cluster Enhancement (TFCE), a strict multiple comparison correction strategy, reached the best balance between family-wise error rate (under 5%) and test-retest reliability/replicability (e.g., 0.68 for test-retest reliability and 0.25 for replicability of amplitude of low-frequency fluctuations (ALFF) for between-subject sex differences, 0.49 for replicability of ALFF for within-subject EOEC differences). Although R-fMRI indices attained moderate reliabilities, they replicated poorly in distinct datasets (replicability < 0.3 for between-subject sex differences, < 0.5 for within-subject EOEC differences). By randomly drawing different sample sizes from a single site, we found reliability, sensitivity and positive predictive value (PPV) rose as sample size increased. Small sample sizes (e.g., < 80 [40 per group]) not only minimized power (sensitivity < 2%), but also decreased the likelihood that significant results reflect "true" effects (PPV < 0.26) in sex differences. Our findings have implications for how to select multiple comparison correction strategies and highlight the importance of sufficiently large sample sizes in R-fMRI studies to enhance reproducibility. Hum Brain Mapp 39:300-318, 2018. © 2017 Wiley Periodicals, Inc.


A highly sensitive fluorescent light-up probe for real-time detection of the endogenous protein target and its antagonism in live cells.
Real-time detection and monitoring of cancer-related biomolecular interactions in live cells are of paramount importance for disease diagnostics and drug screening. Herein, we developed a target-specific fluorescent light-up probe for cellular detection of Mdm2, the key negative regulator of the p53 tumour suppressor protein. Conjugation of a uniquely designed fluorogen (TPECM) with aggregation induced-emission properties, to a specific p53-derived peptide (12.1Pep) targeting Mdm2, yielded a cell-permeable probe (TPECM-12.1Pep) with turn-on fluorescence properties for real-time live cell imaging of Mdm2. This specific light-up probe is almost non-fluorescent in its isolated state but is highly emissive upon binding to Mdm2, enabling quantitative detection of both Mdm2 and its antagonism. Using a model compound (Nutlin-3a), we demonstrate that the as-developed probes can be used to screen p53-Mdm2 inhibiting drug candidates, both in vitro and in cells. Furthermore, the probe activity can be accurately monitored in cells using a fluorescently activated cell sorting machine. These features will expedite research in the areas of drug discovery, clinical diagnostics and fundamental cell biology.


Downregulation of KCNMA1 in mice accelerates auditory hair cells senescence via ferroptosis.
KCNMA1 encodes the K+ potassium channel α-subunit that plays a significant role in the auditory system. Our previous studies indicated that KCNMA1 is associated with age-related hearing loss(AHL). However, the detailed mechanism of KCNMA1 involvement in auditory age-related degradation has not been fully clarified. Therefore, we explored the expression of KCNMA1 in the peripheral auditory of 2-month-old and 12-month-old mice by Western blotting and immunofluorescence. The results of animal experiments showed that KCNMA1 expression was decreased in 12-month-old mice compared with 2-month-old mice, whereas the ferroptosis level was increased. To verify the role of KCNMA1 in AHL, we downregulated KCNMA1 in HEI-OC1 cells by transfecting shRNA. After downregulation, the ferroptosis level was increased and the aging process was accelerated. Furthermore, the aging process was affected by the expression of ferroptosis. In conclusion, these results revealed that KCNMA1 is associated with the aging process in auditory hair cells by regulating ferroptosis, which deepens our understanding of age-related hearing loss.


Functional status and clinical correlates in cognitively impaired community-living older people.
We describe the prevalence of cognitive impairment in a population of community-living older people, its association with functional decline, and degree of comorbidity. In addition, we examined the relationship between different levels of cognitive impairment and mortality. We conducted an observational study of 1787 patients aged 65 years and above with any degree of cognitive impairment. Patient data were collected with the Minimum Data Set for Home Care. More than 50% of patients had some level of cognitive impairment, which correlates with the degree of physical frailty. On the contrary, patients with cognitive impairment appear to have fewer comorbid conditions and are less likely to receive medications than patients with normal cognitive status. In particular, hypertension, congestive heart failure, chronic obstructive pulmonary disease, cancer, diabetes mellitus, and osteoporosis are found more frequently among patients with normal mental status compared with those showing some level of cognitive defects. Yet, more severe cognitive impairment is associated with a higher mortality rate. Demented patients are characterized by a high prevalence of functional disability and by increased mortality. This increased morbidity and mortality rate is associated with a lower prevalence of comorbid clinical conditions and drug use, relative to patients with normal cognitive performance. The present findings support the possibility that severe cognitive impairment has an independent effect on survival.


The genetics of lissencephaly.
Lissencephaly is a spectrum of severe brain malformations caused by the failure of migrating neurons to reach optimal positions in the developing cerebral cortex. Several syndromes associated with lissencephaly have been characterized in recent years. Identification of the genetic basis of these disorders has brought fascinating insights into the mechanisms of brain development, as well as benefits to patients through improved molecular diagnosis and genetic counseling. This review explores the clinical presentation, radiological features, histological findings and molecular basis of lissencephaly with the aim of facilitating the selection and interpretation of gene tests in patients with 'smooth brain' phenotypes.


Longitudinal Changes in Cardiac Structure and Function From Adolescence to Young Adulthood in Participants With Type 2 Diabetes Mellitus: The TODAY Follow-Up Study.
Heart failure is a prominent complication of type 2 diabetes mellitus (T2D). The goal of this study was to provide longitudinal data on cardiac structure and function (and cross-sectional comparison to normal-weight and obese controls without T2D) in individuals followed from adolescence with youth-onset T2D. In the TODAY study (Treatment Options for Type 2 Diabetes Mellitus in Adolescents and Youth), echocardiograms were performed at study years 4 to 5 and 9 to 10. Echocardiograms were also obtained at years 8 to 9 in a control population of age, race/ethnicity, and sex-matched normal-weight and obese individuals without diabetes mellitus. Study outcomes were measures of left ventricular structure and function. The cohort included 411 participants with T2D, 194 obese controls, and 51 normal-weight controls. At follow-up, mean participant age was 23 years, 65% women, 20% non-Hispanic white, 35% non-Hispanic black, and 39% Hispanic. Ejection fraction was <52% in 11.7% of male participants with T2D. Diastolic function declined during follow-up in participants with T2D (mitral valve lateral E/Em increased 0.72±0.12 in women and 0.50±0.17 in men; P<0.01) and was significantly higher than obese controls (women, 6.65±1.89 versus 5.66±1.37; men, 6.15±1.90 versus 5.26±1.31; P<0.0001). Predictors of adverse changes included hypertension, obesity, female sex, Hispanic and non-Hispanic black ethnicity, worse glycemic control, and elevated heart rate. Cardiac structural abnormalities, left ventricular hypertrophy, or concentric geometry, were highest in those with T2D (15.8% versus 5.7% obese versus 0% normal weight). Adverse changes in cardiac structure and function changed significantly from adolescence to early adulthood in participants with youth-onset T2D. Registration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT00081328.


[Limited indication for percutaneous lumbal nucleotomies.].
The aim of our retrospective study is to show how very circumspect we are to use this method. The clinical symptoms of our patients were lumbalgia or lumboischialgia with or without neurological deficiencies. All these patients were resistent to conservative therapies. We accepted for operation only patients who suffered from protrusions and types of prolaps without perforation of the posterior longitudinal ligament. 179 operations on the vertebral discs were performed in our department between 01. 01. 1992 until 31.12.1997. Of this number 165 (92 %) were conservative nucleotomies (63 females and 102 males) and 14 patients (8 %) hat percutane nucleotomies and discos-copies (6 females and 8 males). The average age at the time of operation was 49 years (from 12 to 81 years) with patients who had an open operation, whilst the patients who had percutane nucleotomy had an average age of 42 years (26 to 56 years). The time span between postoperative checkups was on average about 40 months (3 years and 4 months). The shortest postoperative checkup period was 2 months and the longest 71 months (5 years and 11 months). Of the 14 cases mentioned we operated 9 patients in the segment L4/5, 4 patients in the segment L5/S1 and one patient in the segment L3/4. In 7 instances it was the right side that was injured and 7 times it was the left side which was injured. We performed all operations under general anaesthesia, the patient lying prone on the abdomen with lumbal lordosis raised as much as possible. We always used an X-ray picture enlarger on two planes. As far as subjective pain is concerned, an analysis of the findings gives the following results: 100 % improvement 6 patients, 80 % improvement 2 patients, 50 % improvement 3 patients, no improvement 3 patients. Five praeoperative existing motorial deficiencies had completely vanished postoperatively. As to eventual complications, we had one patient suffering from a lesion at the root of L4 on the right side with postoperative Spondylodiscitis L3/4 who was treated in a conservative way. When patient selection for this method is critically considered, the advantage of endoscopic operation lies in minimal invasion and rapid rehabilitation is associated with it.


Mechanosensing and mechanochemical transduction: how is mechanical energy sensed and converted into chemical energy in an extracellular matrix?
Gravity plays a central role in vertebrate development and evolution. Gravitational forces acting on mammalian tissues cause the net muscle forces required for locomotion to be higher on earth than on a body subjected to a microgravitational field. As body mass increases during development, the musculoskeleton must be able to adapt by increasing the size of its functional units. Thus mechanical forces required to do the work (mechanical energy) of locomotion must be sensed by cells and converted into chemical energy (synthesis of new tissue). Extracellular matrices (ECMs) are multicomponent tissues that transduce internal and external mechanical signals into changes in tissue structure and function through a process termed mechanochemical transduction. Under the influence of an external gravitational field, both mineralized and unmineralized vertebrate tissues exhibit internal tensile forces that serve to preserve a synthetic phenotype in the resident cell population. Application of additional external forces alters the balance between the external gravitational force and internal forces acting on resident cells leading to changes in the expression of genes and production of protein that ultimately may alter the exact structure and function of the extracellular matrix. Changes in the equilibrium between internal and external forces acting on ECMs and changes in mechanochemical transduction processes at the cellular level appear to be important mechanisms by which mammals adjust their needs to store, transmit, and dissipate energy that is required during development and for bodily movements. Mechanosensing is postulated to involve many different cellular and extracellular components. Mechanical forces cause direct stretching of protein-cell surface integrin binding sites that occur on all eukaryotic cells. Stress-induced conformational changes in the extracellular matrix may alter integrin structure and lead to activation of several secondary messenger pathways within the cell. Activation of these pathways leads to altered regulation of genes that synthesize and catabolize extracellular matrix proteins as well as to alterations in cell division. Another aspect by which mechanal signals are transduced involves deformation of gap junctions containing calcium-sensitive stretch receptors. Once activated, these channels trigger secondary messenger activation through pathways similar to those involved in integrin-dependent activation and allow cell-to-cell communications between cells with similar and different phenotypes. Another process by which mechanochemical transduction occurs is through the activation of ion channels in the cell membrane. Mechanical forces have been shown to alter cell membrane ion channel permeability associated with Ca(+2) and other ion fluxes. In addition, the application of mechanical forces to cells leads to the activation of growth factor and hormone receptors even in the absence of ligand binding. These are some of the mechanisms that have evolved in vertebrates by which cells respond to changes in external forces that lead to changes in tissue strcture and function.


A truncated and dimeric format of an Affibody library on bacteria enables FACS-mediated isolation of amyloid-beta aggregation inhibitors with subnanomolar affinity.
The amyloid hypothesis suggests that accumulation of amyloid β (Aβ) peptides in the brain is involved in development of Alzheimer's disease. We previously generated a small dimeric affinity protein that inhibited Aβ aggregation by sequestering the aggregation prone parts of the peptide. The affinity protein is originally based on the Affibody scaffold, but is evolved to a distinct interaction mechanism involving complex structural rearrangement in both the Aβ peptide and the affinity proteins upon binding. The aim of this study was to decrease the size of the dimeric affinity protein and significantly improve its affinity for the Aβ peptide to increase its potential as a future therapeutic agent. We combined a rational design approach with combinatorial protein engineering to generate two different affinity maturation libraries. The libraries were displayed on staphylococcal cells and high-affinity Aβ-binding molecules were isolated using flow-cytometric sorting. The best performing candidate binds Aβ with a KD value of around 300 pM, corresponding to a 50-fold improvement in affinity relative to the first-generation binder. The new dimeric Affibody molecule was shown to capture Aβ1-42 peptides from spiked E. coli lysate. Altogether, our results demonstrate successful engineering of this complex binder for increased affinity to the Aβ peptide.


Activation of the DNA-binding ability of latent p53 protein by protein kinase C is abolished by protein kinase CK2.
p53 is one of the most important regulators of cell proliferation and differentiation and of programmed cell death, triggering growth arrest and/or apoptosis in response to different cellular stress signals. The sequence-specific DNA-binding function of p53 protein can be activated by several different stimuli that modulate the C-terminal domain of this protein. The predominant mechanism of activation of p53 sequence-specific DNA binding is phosphorylation at specific sites. For example, phosphorylation of p53 by PKC (protein kinase C) occurs in undamaged cells, resulting in masking of the epitope recognized by monoclonal antibody PAb421, and presumably promotes steady-state levels of p53 activity in cycling cells. In contrast, phosphorylation by cdk2 (cyclin-dependent kinase 2)/cyclin A and by the protein kinase CK2 are both enhanced in DNA-damaged cells. We determined whether one mechanism to account for this mutually exclusive phosphorylation may be that each phosphorylation event prevents modification by the other kinase. We used non-radioactive electrophoretic mobility shift assays to show that C-terminal phosphorylation of p53 protein by cdk2/cyclin A on Ser315 or by PKC on Ser378 can efficiently stimulate p53 binding to DNA in vitro, as well as binding of the monoclonal antibody Bp53-10, which recognizes residues 371-380 in the C-terminus of p53. Phosphorylation of p53 by CK2 on Ser392 induces its DNA-binding activity to a much lower extent than phosphorylation by cdk2/cyclin A or PKC. In addition, phosphorylation by CK2 strongly inhibits PKC-induced activation of p53 DNA binding, while the activation of p53 by cdk2/cyclin A is not affected by CK2. The presence of CK2-mediated phosphorylation promotes PKC binding to its docking site within the p53 oligomerization domain, but decreases phosphorylation by PKC, suggesting that competition between CK2 and PKC does not rely on the inhibition of PKC-p53 complex formation. These results indicate the crucial role of p53 C-terminal phosphorylation in the regulation of its DNA-binding activity, but also suggest that antagonistic relationships exist between different stress signalling pathways.


The case study as a research strategy.
A research strategy seldom used in the caring sciences is the case study. A case study is an empirical in-depth inquiry about an individual, family, group or organization. It is preferable when 'how' and 'why' questions are asked. The case study is mainly used to explain those causal links in real-life intervention that are too complex for either the survey or experimental strategies. Like other research strategies, its design includes questions or propositions, units of analysis, the logic linking the data to the questions or propositions, and the interpretations of the outcomes. A case study can be reported as a single case or as a compilation of a series of cases. In conclusion, a case study is a simple and excellent way for a care professional to present him or herself to the scientific world.


Digital economy impact on inclusive green growth: intermediary and spatial spillover effects in China.
As a new economic form that has emerged from the technological and digital revolution, the digital economy has great benefits to the green development of the economy, protection of the ecological environment, and improvement in social well-being. This study examines whether the digital economy affects the inclusive green growth (IGG). Measurement index system for the digital economy and IGG from 2013 to 2020 in China was constructed and the entropy evaluation method and super-efficiency epsilon-based measure (Super-EBM) model was used to evaluate them. We examined the direct effect, mediation effect, and spatial spillover effect of the digital economy on IGG using the basic regression model, mediating effect model, and the spatial Durbin model. Our results revealed that the development of the digital economy has clear regional differences. The role of digital economy on IGG has positive direct effects and positive spatial spillover effects. The digital economy affects IGG through industrial structure upgrading, so industrial structure upgrading has a significant intermediary effect. By comparing the coefficients, we found that the spatial spillover effects are stronger than the direct effects. Absorbing the spillover of the digital economy in surrounding areas is particularly important for promoting IGG. Heterogeneity analysis revealed that the digital economy is better for IGG and releases more digital dividends in eastern China. Finally, the research conclusions of this paper provide reference for the Chinese government to formulate relevant policies to deepen the integration of digital information technology and IGG, and promote the coordinated development of regional digital economy and IGG.


Bipolarity in a Collaborative Care Model Variation: Detection, Prevalence, and Outcomes.
To assess the prevalence of bipolarity and its impact on clinical course, psychiatric consultants' diagnostic impressions and respective treatment outcomes were examined for patients with depression who were treated in a collaborative care model (CoCM) of psychiatric consultation. Electronic records for 1,476 patients were reviewed for the presence of a mood disorder, which yielded 641 patients with complete data on several measures: the Composite International Diagnostic Interview, version 3.0 (CIDI); a questionnaire eliciting data on non-mania-related markers of bipolar disorder (family history, age of onset, course of illness, response to treatment); consultants' diagnostic impressions; and Patient Health Questionnaire-9 (PHQ-9) scores before and after consultation. Of referred patients, 97% were screened for bipolar disorder. A smooth distribution of scores on the CIDI was observed. Patients were divided into four groups on the basis of their CIDI scores (≥7, positive, or <7, negative) and on the consultant's recorded Impression (positive or negative for bipolarity). Of the study sample, 21% were CIDI positive (≥7), and 35% were Impression positive (sufficient bipolarity to guide treatment recommendations). All groups demonstrated equivalent decreases in PHQ-9 scores in the 6 months since consultation, including the potentially overdiagnosed group (CIDI negative, impression positive), which comprised 22% of the study sample. Universal screening for bipolarity in primary care is feasible in CoCM programs. Interpreting the data dimensionally is logical on the basis of the smooth distribution of CIDI scores. Such screening will yield high rates of bipolar disorder, much higher than previously reported. Offering treatment recommendations based on an impression of bipolarity to patients with negative CIDI results (<7) was not associated with outcomes worse than experienced by all other consultation patients. Multiple explanations of the latter finding are possible, warranting additional study.


Nano-scientific Application of Atomic Force Microscopy in Pathology: from Molecules to Tissues.
The advantages of atomic force microscopy (AFM) in biological research are its high imaging resolution, sensitivity, and ability to operate in physiological conditions. Over the past decades, rigorous studies have been performed to determine the potential applications of AFM techniques in disease diagnosis and prognosis. Many pathological conditions are accompanied by alterations in the morphology, adhesion properties, mechanical compliances, and molecular composition of cells and tissues. The accurate determination of such alterations can be utilized as a diagnostic and prognostic marker. Alteration in cell morphology represents changes in cell structure and membrane proteins induced by pathologic progression of diseases. Mechanical compliances are also modulated by the active rearrangements of cytoskeleton or extracellular matrix triggered by disease pathogenesis. In addition, adhesion is a critical step in the progression of many diseases including infectious and neurodegenerative diseases. Recent advances in AFM techniques have demonstrated their ability to obtain molecular composition as well as topographic information. The quantitative characterization of molecular alteration in biological specimens in terms of disease progression provides a new avenue to understand the underlying mechanisms of disease onset and progression. In this review, we have highlighted the application of diverse AFM techniques in pathological investigations.


Cell therapy of alpha-sarcoglycan null dystrophic mice through intra-arterial delivery of mesoangioblasts.
Preclinical or clinical trials for muscular dystrophies have met with modest success, mainly because of inefficient delivery of viral vectors or donor cells to dystrophic muscles. We report here that intra-arterial delivery of wild-type mesoangioblasts, a class of vessel-associated stem cells, corrects morphologically and functionally the dystrophic phenotype of virtually all downstream muscles in adult immunocompetent alpha-sarcoglycan (alpha-SG) null mice, a model organism for limb-girdle muscular dystrophy. When mesoangioblasts isolated from juvenile dystrophic mice and transduced with a lentiviral vector expressing alpha-SG were injected into the femoral artery of dystrophic mice, they reconstituted skeletal muscle in a manner similar to that seen in wild-type cells. The success of this protocol was mainly due to widespread distribution of donor stem cells through the capillary network, a distinct advantage of this strategy over previous approaches.


Multi-phasic eGFR trajectory during follow up and long-term graft failure after kidney transplantation.
The prevailing assumption is that following kidney transplantation the pattern of kidney function decline is consistent. Nevertheless, numerous factors leading to graft loss may emerge, altering the trajectory of kidney function. In this study, we aim to assess alterations in estimated glomerular filtration rate (eGFR) trajectory over an extended period of follow-up and examine its correlation with graft survival. We calculated eGFR using all creatinine values available from 1-year post transplantation to the end of follow-up. For pattern analysis, we used a piecewise linear model. Nine hundred eighty-eight patients were included in the study. After a median follow-up of 5.2 years, 297 (30.1%) patients had a multi-phasic eGFR trajectory. Change in eGFR trajectory was associated with increased risk for graft failure (HR 7.15, 95% CI 5.17-9.89, p < .001), longer follow-up time, younger age, longer cold ischemia time, high prevalence of acute rejection, longer hospitalization and a lower initial eGFR. Of the 988 patients included in the study, 494 (50.0%) had a mono-phasic stable trajectory, 197 (19.9%) had a mono-phasic decreasing trajectory, 184 (18.6%) had bi-phasic decreasing trajectory (initial stability and then decline, 46(4.7%) had a bi-phasic stabilized (initial decline and then stabilization) and 67(6.8%) had a more complex trajectory (tri-phasic). Out of the total 144 patients who experienced graft loss, the predominant pattern was a bi-phasic decline characterized by a bi-linear trajectory (66 events, 45.8%). Changes in eGFR trajectory during long-term follow-up can serve as a valuable tool for assessing the underlying mechanisms contributing to graft loss.


Application of extracorporeal shock wave lithotripter (ECSWL) in orthopaedics. II. Dose-response and pressure distribution measurements.
In order to apply the extracorporeal shock wave lithotripter (ECSWL) technique to the loosening of the bone-cement interface for the extraction of the cement during revision arthroplasty it is essential to know the dose-response characteristics. The present study shows that the number of shocks needed to break the interface between a 2- and 6-mm-thick bovine femoral bone and bone cement is similar to the fatigue behavior of a material, that is, Log(N) = C(kV) + D, where N is the number of shock impulses, kV is the power setting of the lithotripter machine in kilovolts, and C and D are constants. Iso-pressure distribution of the traveling shock wave front through a simulated bone in a Plexiglass tube using Fuji pressure film showed quantitative pressure contours from which one can understand the effective area of shock wave and its distribution. The most effective area of the shock wave was about 1.5 cm in diameter at 23 and 25 kV with pressure at least 7.0 MPa which is more than sufficient to break the bone-cement interface in tension.


Health state utilities and quality of life in patients with hepatitis B.
The effect of chronic hepatitis B (CHB) infection on health-related quality of life (HRQoL) and health state utilities has not been well characterized. To measure utility scores and HRQoL across disease states associated with CHB infection. Patients attending four tertiary care clinics for CHB were approached between July 2007 and March 2009. Respondents completed version 2 of the Short-Form 36 Health Survey, the EQ5D, a visual analogue scale, the Health Utilities Index Mark 3, standard gamble, and demographics and risk factor surveys in English, Cantonese or Mandarin. Charts were reviewed to determine disease stage and comorbidities. A total of 433 patients were studied: 294 had no cirrhosis; 79 had compensated cirrhosis; seven had decompensated cirrhosis; 23 had hepatocellular carcinoma; and 30 had received a liver transplant. The mean standard gamble utilities for these disease states were 0.89, 0.87, 0.82, 0.84 and 0.86, respectively. HRQoL scores in noncirrhotic patients were similar to those of the general population. Scores of patients with compensated cirrhosis were not significantly lower; however, patients with decompensated cirrhosis and hepatocellular carcinoma had significantly lower HRQoL scores compared with noncirrhotic patients (P<0.05). Similar scores were observed among patients on and off oral antiviral treatment. Post-liver transplant patients had a higher HRQoL than patients with decompensated cirrhosis. Age, number of comorbidities and relationship status were significantly associated with HRQoL scores. HRQoL in CHB patients is only impaired in the later stages of liver disease. Neither CHB infection nor antiviral treatment is associated with a lower quality of life.


Targeting tRNA-synthetase interactions towards novel therapeutic discovery against eukaryotic pathogens.
The development of chemotherapies against eukaryotic pathogens is especially challenging because of both the evolutionary conservation of drug targets between host and parasite, and the evolution of strain-dependent drug resistance. There is a strong need for new nontoxic drugs with broad-spectrum activity against trypanosome parasites such as Leishmania and Trypanosoma. A relatively untested approach is to target macromolecular interactions in parasites rather than small molecular interactions, under the hypothesis that the features specifying macromolecular interactions diverge more rapidly through coevolution. We computed tRNA Class-Informative Features in humans and independently in eight distinct clades of trypanosomes, identifying parasite-specific informative features, including base pairs and base mis-pairs, that are broadly conserved over approximately 250 million years of trypanosome evolution. Validating these observations, we demonstrated biochemically that tRNA:aminoacyl-tRNA synthetase (aaRS) interactions are a promising target for anti-trypanosomal drug discovery. From a marine natural products extract library, we identified several fractions with inhibitory activity toward Leishmania major alanyl-tRNA synthetase (AlaRS) but no activity against the human homolog. These marine natural products extracts showed cross-reactivity towards Trypanosoma cruzi AlaRS indicating the broad-spectrum potential of our network predictions. We also identified Leishmania major threonyl-tRNA synthetase (ThrRS) inhibitors from the same library. We discuss why chemotherapies targeting multiple aaRSs should be less prone to the evolution of resistance than monotherapeutic or synergistic combination chemotherapies targeting only one aaRS.


Single-center 10-year experience in treating patients with vascular tinnitus: diagnostic approaches and treatment outcomes.
Vascular tinnitus is the most common form of pulsatile tinnitus, particularly when the tinnitus corresponds with the pulse of patients. In this study, we reviewed the 10-year clinical data on vascular tinnitus of our tinnitus clinic to investigate the frequency of the underlying etiologies, to introduce a diagnostic protocol, and to evaluate the treatment outcomes. We retrospectively collected the data of 57 patients who were diagnosed as vascular tinnitus between April 2001 and December 2011. Careful history taking, otoscopy, thorough physical examinations, audiometry, laboratory tests, as well as radiologic examinations were performed according to our diagnostic protocol to find the origin of pulsatile tinnitus. Treatment options were individualized based on the specific etiology, and the outcomes were assessed using patient's subjective reports at the follow-up interviews. High jugular bulb was the most common cause (47.4%) of vascular tinnitus, and venous hum was the next (17.5%). Dural arteriovenous fistula, intracranial aneurysm, atherosclerotic carotid artery disease, and hypertension were less common causes. Vascular tinnitus was alleviated in most patients after the appropriate treatment: surgical intervention, tinnitus retraining therapy, reassurance, and medications. Vascular tinnitus can be successfully diagnosed by the regular use of the suggested protocol. Many patients with vascular tinnitus have treatable underlying etiologies. Treatment of those etiologies or at least counseling about the tinnitus itself can benefit the patients with troublesome vascular tinnitus.


Secondary Evolve and Resequencing: An Experimental Confirmation of Putative Selection Targets without Phenotyping.
Evolve and resequencing (E&R) studies investigate the genomic responses of adaptation during experimental evolution. Because replicate populations evolve in the same controlled environment, consistent responses to selection across replicates are frequently used to identify reliable candidate regions that underlie adaptation to a new environment. However, recent work demonstrated that selection signatures can be restricted to one or a few replicate(s) only. These selection signatures frequently have weak statistical support, and given the difficulties of functional validation, additional evidence is needed before considering them as candidates for functional analysis. Here, we introduce an experimental procedure to validate candidate loci with weak or replicate-specific selection signature(s). Crossing an evolved population from a primary E&R experiment to the ancestral founder population reduces the frequency of candidate alleles that have reached a high frequency. We hypothesize that genuine selection targets will experience a repeatable frequency increase after the mixing with the ancestral founders if they are exposed to the same environment (secondary E&R experiment). Using this approach, we successfully validate two overlapping selection targets, which showed a mutually exclusive selection signature in a primary E&R experiment of Drosophila simulans adapting to a novel temperature regime. We conclude that secondary E&R experiments provide a reliable confirmation of selection signatures that either are not replicated or show only a low statistical significance in a primary E&R experiment unless epistatic interactions predominate. Such experiments are particularly helpful to prioritize candidate loci for time-consuming functional follow-up investigations.


A scanning electron microscopic study of the dorsal surface of the human tongue.
To study the dorsal surface of the human tongue using a scanning electron microscopy (SEM), tissue specimens were taken from the anterior part of the tongues of 15 individuals aged from 21- to 28-years-old. The formalin-fixed samples were processed routinely for SEM. With SEM the surface of the normal tongue mucosa was shown to be rather evenly covered by filiform papillae, with some fungiform papillae scattered among them. Filiform papillae consisted of two parts: the body and hairs. The mucosal surface of the body was smooth; the squamous epithelial cells were polygonal, and their boundaries were prominent. On the surface of the superficial epithelial cells were parallel or branching microplicae. Each filiform papilla had 6-10 hairs, which were scaled and covered by an extensive plaque of microorganism. The upper surface of the fungiform papillae was smooth; only a few desquamating cells were seen. The superficial cells had a pitted appearance and cell boundaries overlapped. Taste pores, up to 3 pores in a single papilla, were found on the upper surface. Desquamation was more pronounced on the base of the fungiform papillae than on the upper surface. In almost all fungiform papillae some hairs protruded from the base. Parallel microplicae were found on the surface of the superficial cells of the base. The structure and function of the human tongue, as well as the microplicae of its superficial cells, are compared to those of various species of animals.


Biomarkers in inflammatory bowel disease: current practices and recent advances.
Crohn's disease and ulcerative colitis represent the two main forms of the idiopathic chronic inflammatory bowel diseases (IBD). Currently available blood and stool based biomarkers provide reproducible, quantitative tools that can complement clinical assessment to aid clinicians in IBD diagnosis and management. C-reactive protein and fecal based leukocyte markers can help the clinician distinguish IBD from noninflammatory diarrhea and assess disease activity. The ability to differentiate between forms of IBD and predict risk for disease complications is specific to serologic tests including antibodies against Saccharomyces cerevisiae and perinuclear antineutrophil cytoplasmic proteins. Advances in genomic, proteomic, and metabolomic array based technologies are facilitating the development of new biomarkers for IBD. The discovery of novel biomarkers, which can correlate with mucosal healing or predict long-term disease course has the potential to significantly improve patient care. This article reviews the uses and limitations of currently available biomarkers and highlights recent advances in IBD biomarker discovery.