Inexpensive, rapid and convenient PCR-minigel SSCP protocol for polymorphisms and mutations analyses of LDL receptor gene.
Hypercholesterolemia has been recognized as a major risk factor of atherosclerosis and coronary artery disease. The elevation in plasma low density lipoprotein (LDL) cholesterol is frequently due to genetic alteration at the genetic locus specifying the LDL receptors, leading to defective catabolism of LDL. In order to facilitate the molecular diagnosis of LDL receptor disorder, single strand conformation polymorphism (SSCP) analysis of polymerase chain reaction (PCR) amplified genomic DNA fragments has become a simple and sensitive screening method for identification of DNA polymorphisms and mutations in LDL receptor gene prior to DNA sequencing. In addition, SSCP patterns can be detected by silver staining to avoid hazardous radioactive material or other costly nonradioactive detection techniques. However, the original SSCP protocol is generally large-formatted, which is both time and reagents consuming as well as cumbersome. Minigel SSCP protocols have thus been devised but they involve, although commercially available, costly precast gels. We describe here a nonradioactive PCR-minigel SSCP protocol which is sensitive, inexpensive, rapid, reproducible and manually convenient. The results in this study demonstrate that minigel-SSCP (gel size: 10 cm x 7.3 cm x 0.075 cm) can detect conformation polymorphisms in PCR-fragments with a comparative sensitivity to large gel SSCP (gel size: 30 cm x 40 cm x 0.04 cm) as exemplified by the SSCP analyses of exon 13 of the LDL receptor gene. For minigel SSCP, the reagents for gel components and silver staining are reduced approximately 9 times and 10 times, respectively. For electrophoresis, electrical power is also reduced 10 times. This improved technique can become routinely used for molecular diagnosis of LDL receptor defect as well as for other genetic disorders.


Mindfulness Training, Implicit Bias, and Force Response Decision-making.
The goal of this study was to assess the preliminary efficacy of a Mindfulness-Based Training (Mindfulness-based Resilience Training; MBRT) in improving weapon identification among Law Enforcement Officers (LEOs). Participants (N = 61) were randomly assigned to either MBRT or a no-intervention control group (NIC). A self-report questionnaire assessing mindfulness and a computerized measure assessing implicit stereotype reliance were administered at baseline, post-training, and three months following intervention completion. We also collected information about meditation practice outside of the training for LEOs in the MBRT group. Inferential analyses yielded improvements in mindfulness in the MBRT group compared to NIC. Analyses did not provide evidence for implicit stereotype reliance at baseline and therefore did not yield a significant impact of MBRT versus NIC on implicit stereotype reliance, ps > .05; however, participants across both conditions exerted more control when responding to Black male targets compared to White male targets, F(1,74) = 3.98, p = .05, 95% CI [-.05, -.01], d = .36. Our results do not provide evidence for the impact of MBRT on weapon identification, but do suggest that LEOs exerted more effort when responding to images of Black males compared to White males. We discuss recommendations for future clinical trials assessing implicit stereotype reliance, viz., that researchers utilize measures more sensitive to a wider range of LEO samples and with higher ecological validity, and we discuss potential reasons why our results do not align with past research.


Beam-surface scattering studies of the individual and combined effects of VUV radiation and hyperthermal O, O2, or Ar on FEP Teflon surfaces.
Beam-surface scattering experiments were used to probe products that scattered from FEP Teflon surfaces during bombardment by various combinations of atomic and molecular oxygen, Ar atoms, and vacuum ultraviolet (VUV) light. A laser-breakdown source was used to create hyperthermal (translational energies in the range 4-13 eV) beams of argon and atomic/molecular oxygen. The average incidence energy of these beams was tunable and was controlled precisely with a synchronized chopper wheel. A filtered deuterium lamp provided a source of VUV light in a narrow-wavelength range centered at 161 nm. Volatile products that exited the surfaces were monitored with a rotatable mass spectrometer detector. Hyperthermal O atoms with average translational energies above approximately 4 eV may react directly with a pristine FEP Teflon surface, and the reactivity appears to increase with the translational energy of the incident O atoms. VUV light or highly energetic collisions of O2 or Ar may break chemical bonds and lead to the ejection of volatile products; the ejection of volatile products is enhanced when the surface is subjected to VUV light and energetic collisions simultaneously. Exposure to VUV light or to hyperthermal O2 or Ar may increase the reactivity of an FEP Teflon surface to O atoms.


Soil Mineralogy as Factor in Spread of Fusarium Wilt of Banana.
A correlation is established between the spread of Fusarium wilt of banana and soil mineralogy. Montmorillonoid-type clay minerals occur in all soils in which disease spread is slow, but, with the exception of two soils, this group of minerals is absent in soils in which it is rapid.


RNAi-mediated suppression of constitutive pulmonary gene expression by small interfering RNA in mice.
The ability of synthetic small interfering RNA (siRNA) to silence gene expression makes it a useful tool in biomedical research. However, effective and non-toxic functional siRNA delivery to mouse lungs in vivo is still a key challenge, and regulation of constitutively expressed genes is poorly characterized. Following in vitro validation of siRNA molecules, naked, stabilized siRNA (AtuRNAi) was applied intranasally (i.n.) by droplets or intratracheally (i.t.) by MicroSprayer in female C57BL/6 mice. Distribution of Cy3-tagged siRNAs was examined. Pulmonary expression of ubiquitously (lamin B1) or cell-specific (E-cadherin, VE-cadherin), constitutive genes was analysed by TaqMan-realtime-PCR. Further, formulated lipoplex-siRNA, which has enhanced transfection efficiency, was applied i.t. or intravenously (i.v.). Single i.t. as compared to i.n. application of unformulated siRNA resulted in higher delivery efficiency and homogenous pulmonary distribution. After inhalation of target-specific siRNA, reduction of epithelial E-cadherin by 21%, but no significant reduction of endothelial VE-cadherin or ubiquitously expressed lamin B1 was observed. Pharmacokinetic analysis revealed rapid transfer of intact siRNA molecules into the vascular system and accumulation in the kidneys, calling lung specificity into question. I.t. application of lipoplex-siRNA evoked inflammation. In contrast, i.v. application of lipoplex-siRNA specifically reduced expression of VE-cadherin mRNA by about 50% in lungs without evoking lung cellular influx. In conclusion, sufficient pulmonary distribution of aerosolized siRNA was attained in mice by MicroSprayer, however development of appropriate siRNA carriers is highly desirable to improve lung-specific functional inhalative siRNA delivery. Pulmonary knockdown of constitutive endothelial targets by 50% was achieved by i.v. application of lipoplex-siRNA.


GBF1- and ACBD3-independent recruitment of PI4KIIIβ to replication sites by rhinovirus 3A proteins.
PI4KIIIβ recruitment to Golgi membranes relies on GBF1/Arf and ACBD3. Enteroviruses such as poliovirus and coxsackievirus recruit PI4KIIIβ to their replication sites via their 3A proteins. Here, we show that human rhinovirus (HRV) 3A also recruited PI4KIIIβ to replication sites. Unlike other enterovirus 3A proteins, HRV 3A failed to bind GBF1. Although HRV 3A was previously shown to interact with ACBD3, our data suggest that PI4KIIIβ recruitment occurred independently of both GBF1 and ACBD3.


Localisation of P2X2 receptor subunit immunoreactivity on nitric oxide synthase expressing neurones in the brain stem and hypothalamus of the rat: a fluorescence immunohistochemical study.
A large body of evidence suggests that nitric oxide (NO) and ATP act as neurotransmitters in the regulatory mechanisms concerning several autonomic functions at the level of both the hypothalamus and the brain stem. In the present study, we investigated whether neuronal NO synthase containing neurones also express P2X(2) receptor subunit of the ATP-gated ion channel via double-labelling fluorescence immunohistochemistry. Our data demonstrate that a high percentage of neuronal NO synthase-immunoreactive neurones are also P2X(2)-immunoreactive in the rostral ventrolateral medulla (98%) and supraoptic nucleus of the hypothalamus (92%). Significant numbers of neuronal NO synthase-immunoreactive neurones are also P2X(2)-immunoreactive in the subpostremal (48%) and commissural (65%) subdivisions of the nucleus tractus solitarius. In the caudal ventrolateral medulla and raphe obscurus, 96% and 89%, respectively, of neuronal NO synthase containing neurones also express P2X(2) receptor subunit. In contrast to the supraoptic nucleus, there was a lower percentage of co-localisation between NO synthase and P2X(2) receptor subunit in the paraventricular nucleus of the hypothalamus. In summary, this study demonstrates for the first time that there is a widespread co-localisation of neuronal NO synthase and P2X(2) receptor subunit in the hypothalamus and brain stem of the rat. Further studies are required to elucidate whether NO and ATP functionally interact within the hypothalamus and the brain stem.


Quantum-classical dynamics of scattering processes in adiabatic and diabatic representations.
We demonstrate the workability of a TDDVR based [J. Chem. Phys. 118, 5302 (2003)], novel quantum-classical approach, for simulating scattering processes on a quasi-Jahn-Teller model [J. Chem. Phys. 105, 9141 (1996)] surface. The formulation introduces a set of DVR grid points defined by the Hermite part of the basis set in each dimension and allows the movement of grid points around the central trajectory. With enough trajectories (grid points), the method converges to the exact quantum formulation whereas with only one grid point, we recover the conventional molecular dynamics approach. The time-dependent Schrodinger equation and classical equations of motion are solved self-consistently and electronic transitions are allowed anywhere in the configuration space among any number of coupled states. Quantum-classical calculations are performed on diabatic surfaces (two and three) to reveal the effects of symmetry on inelastic and reactive state-to-state transition probabilities, along with calculations on an adiabatic surface with ordinary Born-Oppenheimer approximation. Excellent agreement between TDDVR and DVR results is obtained in both the representations.


Toxic anterior segment syndrome following trabeculectomy with mitomycin C.
Toxic anterior segment (TASS) is a rare acute sterile anterior segment inflammation that typically develops within 12 to 24 hours after an anterior segment surgery. The purpose of this case report is to alert surgeons to the possibility of this complication following any anterior segment surgery, including trabeculectomy, and to highlight the possible etiologies and measures to prevent it. A 58-year-old male glaucoma patient was initially managed medically for primary open angle glaucoma with antiglaucoma medications. There was rapidly progressive glaucomatous optic nerve damage in his left eye within the following year, despite the use of antiglaucoma medications, hence the need for trabeculectomy. The post-operative condition of the patient's eye was stormy with diffuse limbus-to-limbus corneal edema and profound Descemet's membrane folds, among other features of TASS, with associated deteriorating visual acuity. Although there is no documented report of TASS following trabeculectomy with mitomycin C, surgeons should be alerted to this possibility. Preventive measures include extreme care to avoid errors while preparing and administering diluted solutions, especially medications that are administered into the intracameral space.


An unusual case of orthopnea.
Bilateral diaphragmatic paralysis is a known cause of respiratory failure. Diagnosis can be difficult, particularly in the acute setting. We present the case of a gentleman diagnosed with bilateral diaphragmatic paralysis secondary to phrenic neuropathy in the setting of cervical spondylosis.


At what costs will screening with CT colonography be competitive? A cost-effectiveness approach.
The costs of computed tomographic colonography (CTC) are not yet established for screening use. In our study, we estimated the threshold costs for which CTC screening would be a cost-effective alternative to colonoscopy for colorectal cancer (CRC) screening in the general population. We used the MISCAN-colon microsimulation model to estimate the costs and life-years gained of screening persons aged 50-80 years for 4 screening strategies: (i) optical colonoscopy; and CTC with referral to optical colonoscopy of (ii) any suspected polyp; (iii) a suspected polyp >or=6 mm and (iv) a suspected polyp >or=10 mm. For each of the 4 strategies, screen intervals of 5, 10, 15 and 20 years were considered. Subsequently, for each CTC strategy and interval, the threshold costs of CTC were calculated. We performed a sensitivity analysis to assess the effect of uncertain model parameters on the threshold costs. With equal costs ($662), optical colonoscopy dominated CTC screening. For CTC to gain similar life-years as colonoscopy screening every 10 years, it should be offered every 5 years with referral of polyps >or=6 mm. For this strategy to be as cost-effective as colonoscopy screening, the costs must not exceed $285 or 43% of colonoscopy costs (range in sensitivity analysis: 39-47%). With 25% higher adherence than colonoscopy, CTC threshold costs could be 71% of colonoscopy costs. Our estimate of 43% is considerably lower than previous estimates in literature, because previous studies only compared CTC screening to 10-yearly colonoscopy, where we compared to different intervals of colonoscopy screening.


Attitudes Expressed in Online Comments about Environmental Factors in the Tourism Sector: An Exploratory Study.
The object of this exploratory study is to identify the positive, neutral and negative environment factors that affect users who visit Spanish hotels in order to help the hotel managers decide how to improve the quality of the services provided. To carry out the research a Sentiment Analysis was initially performed, grouping the sample of tweets (n = 14459) according to the feelings shown and then a textual analysis was used to identify the key environment factors in these feelings using the qualitative analysis software Nvivo (QSR International, Melbourne, Australia). The results of the exploratory study present the key environment factors that affect the users experience when visiting hotels in Spain, such as actions that support local traditions and products, the maintenance of rural areas respecting the local environment and nature, or respecting air quality in the areas where hotels have facilities and offer services. The conclusions of the research can help hotels improve their services and the impact on the environment, as well as improving the visitors experience based on the positive, neutral and negative environment factors which the visitors themselves identified.


Mechanisms of Jak/STAT signaling in immunity and disease.
More than two decades ago, experiments on the antiviral mechanisms of IFNs led to the discovery of JAKs and their downstream effectors, the STAT proteins. This pathway has since become a paradigm for membrane-to-nucleus signaling and explains how a broad range of soluble factors, including cytokines and hormones, mediate their diverse functions. Jak/STAT research has not only impacted basic science, particularly in the context of intercellular communication and cell-extrinsic control of gene expression, it also has become a prototype for transition from bench to bedside, culminating in the development and clinical implementation of pathway-specific therapeutics. This brief review synthesizes our current understanding of Jak/STAT biology while taking stock of the lessons learned and the challenges that lie ahead.


[Evaluation of anastomotic effect with esophagogastrostomy covered by tongue-like sero-muscular flap of gastric wall in 1515 cases].
Esophagogastrostomy covered by tongue-like sero-muscular flap of gastric wall has already been performed in 1515 cases with esophageal cancer of middle or lower third segment over the last three years. All cases were of squamous cell carcinoma. The same anastomotic procedure was done in all cases by authors in different hospitals. In this series the operative mortality was 0.07%, there were 5 anastomotic leaks (0.33%), and no anastomotic stricture and reflux esophagitis happened within 6 months to 3 years after surgery. The operative procedure is described again in detail. The authors believe that this new anastomotic procedure is satisfactory and prospectively beneficial for improving the therapeutic effect and patients' living quality after operation.


Synthesis and crystal structure of a new hexagonal perovskite 7H-Ba7Li1.75Mn3.5O15.75 with Mn(4+)/Mn(5+) charge distribution.
Ba7Li1.75Mn3.5O15.75 is a new hexagonal perovskite whose crystal structure has elements typical for the layered hexagonal perovskites and quasi-one-dimensional oxides, hence representing a new polytype. It has been synthesized via a solid-state microwave route. The crystal structure was solved using a combination of X-ray and neutron diffraction data, which show that Ba7Li1.75Mn3.5O15.75 crystallizes in a hexagonal unit cell with parameters a = 5.66274(2) Å and c = 16.7467(1) Å (V = 465.063(4) Å(3)), with one formula unit, and can be described as columns of face-shared octahedra occupied by Mn(4+) and Li(+) cations and vacancies along the c axis separated in the ab plane by barium atoms. Every sixth layer, the coordination of Mn(5+) and Li(+) changes to tetrahedral. Additional local ordering of manganese and lithium atoms among cationic sites leading to the formation of a rhombohedral supercell has been observed by scanning transmission electron microscopy.


[CHRONIC Q FEVER IN PATIENTS AT THE HILLEL YAFFE MEDICAL CENTER - THREE YEARS OF FOLLOW UP].
Q fever is an acute zoonotic infection, which in some cases is complicated by a chronic disease. Diagnosis is based on serology, and in patients with a chronic disease, the source of infection must be investigated. To describe patients with chronic Q fever, who were treated at the Hillel Yaffe Medical Center: risk factors, course of the disease, and serological findings. This was an observational study; patients with chronic Q fever who were treated in the Infectious Diseases Clinic during the period 5/2015 - 1/2018 were included. The diagnosis was based on clinical findings and results of phase 1 IgG ≥ 800. Clinical, laboratory and imaging data from diagnosis to the end of treatment were collected. Sixteen patients were included in the study; all these patients were treated with antibiotics, and three also underwent operations. Risk factors for a chronic infection were a significant valvular disease in 11 patients (69%) and vascular diseases in five (31%). Trans-esophageal echocardiogram (TEE) was performed in 13 patients (81%), and a 18F-FDG/PET-CT was performed in eight patients (50%). The source of infection was found in seven patients, four with endocarditis and three with vascular infection. Endocarditis was more common than vascular infection. In 56% of the patients, the source of the infection was not found. We presented patients with chronic Q fever who were treated in a unique clinic in Israel. Diagnosing the source of the infection is challenging; the increasing use of 18F-FDG/PET-CT allowed accurate diagnosis in some patients in which TEE results were negative.


Regulation of UDP-glucuronosyltransferase 1A1 expression and activity by microRNA 491-3p.
The UDP-glucuronosyltransferase (UGT) 1A enzymes are involved in the phase II metabolism of many important endogenous and exogenous compounds. The nine UGT1A isoforms exhibit high interindividual differences in expression, but their epigenetic regulation is not well understood. The purpose of the present study was to examine microRNA (miRNA) regulation of hepatic UGT1A enzymes and determine whether or not that regulation impacts enzymatic activity. In silico analysis identified miRNA 491-3p (miR-491-3p) as a potential regulator of the UGT1A gene family via binding to the shared UGT1A 3'-untranslated region common to all UGT1A enzymes. Transfection of miR-491-3p mimic into HuH-7 cells significantly repressed UGT1A1 (P < 0.001), UGT1A3 (P < 0.05), and UGT1A6 (P < 0.05) mRNA levels. For UGT1A1, this repression correlated with significantly reduced metabolism of raloxifene into raloxifene-6-glucuronide (ral-6-gluc; P < 0.01) and raloxifene-4'-glucuronide (ral-4'-gluc; P < 0.01). In HuH-7 cells with repressed miR-491-3p expression, there was a significant increase (~80%; P < 0.01) in UGT1A1 mRNA and a corresponding increase in glucuronidation of raloxifene into ral-6-gluc (50%; P < 0.05) and ral-4'-gluc (22%; P < 0.01). Knockdown of endogenous miR-491-3p in HepG2 cells did not significantly alter UGT1A1 mRNA levels but did increase the formation of ral-6-gluc (50%; P < 0.05) and ral-4'-gluc (34%; P < 0.001). A significant inverse correlation between miR-491-3p expression and both UGT1A3 (P < 0.05) and UGT1A6 (P < 0.01) mRNA levels was observed in a panel of normal human liver specimens, with a significant (P < 0.05) increase in UGT1A3 and UGT1A6 mRNA levels observed in miR-491-3p nonexpressing versus expressing liver specimens. These results suggest that miR-491-3p is an important factor in regulating the expression of UGT1A enzymes in vivo.


Magnetic resonance angiography of pulmonary veins: implications for catheter ablation of atrial fibrillation.
Catheter ablation of atrial fibrillation (AF) requires exact anatomical information about pulmonary venous (PV) ostia. In this study, anatomy of pulmonary veins (PVs) was assessed using three-dimensional (3D) reconstructions of magnetic resonance angiography (MRA). Contrast-enhanced MRA of the PVs was performed in 40 patients (mean age 53 +/- 9 years) with paroxysmal (30 patients) or persistent (10 patients) AF, scheduled for circumferential ablation around PV ostia. PV ostial anatomy and diameters were evaluated from multiplanar reconstructions and compared with 3D reconstructions. Thirty (75%) patients presented with a common left-sided antrum (21 short and 9 long trunk), while additional PVs were found on right side in 23%. PV ostia were oblong in shape (mean diameters 17.0 +/- 4.3 vs 10.5 +/- 2.5 mm by two-dimensional (2D) measurements, and 20.8 +/- 7.6 mm vs 12.9 +/- 3.3 mm by 3D reconstruction, in long and short axis, respectively). There was a correlation between measurements obtained from 2D and 3D images, although 3D imaging provided slightly larger diameters. MRA with 3D reconstructions is an important technique for preprocedural assessment of PVs that allows full understanding of their anatomy and size. This information may be important for selection of appropriate tools.


Measurement of kallikrein activity in urine of rats and man using a chromogenic tripeptide substrate. Validation of the amidolytic assay by means of bradykinin radioimmunoassay.
Urinary kallikrein activity in rats and man was measured by an amidolytic assay using the chromogenic tripeptide D-valyl-leucyl-arginine-p-nitroanilide (S 2266). The sensitivity of the assay permits the measurement of 3.5 to 250 mKU of purified porcine pancreatic kallikrein. The intraassay coefficient of variation was 1.6 +/- 0.3% and the interassay coefficient of variation 7.4 +/- 0.9%. No changes in kallikrein activity was found in urine samples kept for 7 days at 4 degrees C for 14 months at -20 degrees C. Repeated freezing and thawing reduced kallikrein activity. Kallikrein activity of the same urine samples was also estimated by means of a bradykinin radioimmunoassay, which determines the kinins released during incubation of urine with partially purified dog kininogen (kininogenase activity). The urinary kallikrein activity assayed by means of these two methods showed a high correlation with coefficients of r = 0.9134 for rat urine (p less than 0.0001) and r = 0.8002 for human urine (p less than 0.001).


Ferrocene-Supported Compartmental Ligands for the Assembly of 3d/4f Complexes.
Site-specific coordination ligands, also known as compartmental ligands, have been used for the preparation of heterometallic complexes. These ligands, by virtue of possessing specific binding sites, can encapsulate different metal ions in their coordination pockets. Such compartmental ligands have been widely used for the preparation of heterometallic 3d/4f complexes which have applications in molecular magnetism. This Review summarizes our efforts in the use of ferrocene-based compartmental ligands for the preparation of heterometallic 3d/4f complexes, some of which are single-molecule or single-ion magnets.


Mesoscale diffusion magnetic resonance imaging of the ex vivo human hippocampus.
Mesoscale diffusion magnetic resonance imaging (MRI) endeavors to bridge the gap between macroscopic white matter tractography and microscopic studies investigating the cytoarchitecture of human brain tissue. To ensure a robust measurement of diffusion at the mesoscale, acquisition parameters were arrayed to investigate their effects on scalar indices (mean, radial, axial diffusivity, and fractional anisotropy) and streamlines (i.e., graphical representation of axonal tracts) in hippocampal layers. A mesoscale resolution afforded segementation of the pyramidal cell layer (CA1-4), the dentate gyrus, as well as stratum moleculare, radiatum, and oriens. Using ex vivo samples, surgically excised from patients with intractable epilepsy (n = 3), we found that shorter diffusion times (23.7 ms) with a b-value of 4,000 s/mm2 were advantageous at the mesoscale, providing a compromise between mean diffusivity and fractional anisotropy measurements. Spatial resolution and sample orientation exerted a major effect on tractography, whereas the number of diffusion gradient encoding directions minimally affected scalar indices and streamline density. A sample temperature of 15°C provided a compromise between increasing signal-to-noise ratio and increasing the diffusion properties of the tissue. Optimization of the acquisition afforded a system's view of intra- and extra-hippocampal connections. Tractography reflected histological boundaries of hippocampal layers. Individual layer connectivity was visualized, as well as streamlines emanating from individual sub-fields. The perforant path, subiculum and angular bundle demonstrated extra-hippocampal connections. Histology of the samples confirmed individual cell layers corresponding to ROIs defined on MR images. We anticipate that this ex vivo mesoscale imaging will yield novel insights into human hippocampal connectivity.


Validation of the diagnostic utility of D-dimer measurement in patients with acute aortic syndrome.
The objective of this study was to evaluate the validity of D-dimer measurements for the diagnosis of acute aortic syndrome in patients admitted to hospital with acute chest pain. A retrospective observational study design was used. Consecutive patients ( n=887) admitted to a tertiary hospital with acute chest pain (acute aortic syndrome, 123; acute pulmonary embolism, 29; and other disease, 735) from the emergency department between January 2011 and April 2014 were assessed to validate the diagnostic value of D-dimer measurements. The D-dimer level was significantly increased in patients with acute aortic syndrome (median (interquartile range) 4.9 (2.0-17.4) µg/ml) compared with control patients (median (interquartile range) 0.6 (0.3-1.4) µg/ml; p<0.001). At a cut-off point of 0.5 μg/ml, the sensitivity for acute aortic syndrome was 0.97 (95% confidence interval 0.92-0.99) and was similar to that for acute pulmonary embolism (0.97 (0.82-0.99)). The age-adjusted D-dimer cut-off point, defined as age × 0.01 μg/ml in patients ⩾50 years, successfully reduced the number of false-positive diagnoses by 13%, while still retaining a high sensitivity (0.96 (0.91-0.99)). The five false-negative diagnoses of acute aortic syndrome included three patients with intramural haematoma, one patient with a penetrating aortic ulcer and one patient with an impending aortic rupture. A combination of probability assessment and the D-dimer approach reduced the number of false-negatives from five patients to two patients. This study demonstrated that the D-dimer test can distinguish acute aortic syndrome from other diseases presenting with acute chest pain with high sensitivity and modest specificity. Using the D-dimer approach presents limitations with some subtypes of acute aortic syndrome, such as intramural haematoma.


[Medullary thyroid carcinoma: the comparison of the hereditary and sporadic types of cancer].
The assessment of frequency and type of mutation and differences in prognosis between sporadic and hereditary type of medullary thyroid carcinoma (MTC), based on own DNA analysis, was performed. The group of 190 persons with hereditary MTC or asymptomatic mutation carriers was analyzed. Patients with sporadic MTC without RET gene mutation were included into control group (708 persons). The recognition of MTC type was based on assessment of family history, physical examination and genetic analysis. The family history consisted of information about MTC, pheochromocytoma and other neoplasms and hyperparathyroidism in relatives. The mutations located in codon 634 of exon 11 were the most often (43% of all mutations and 49% of mutations in syndrome MEN 2A/FMTC). The age of diagnosis was ranged between 7 and 71 years (mean age: 39 +/- 15.2 years, median age: 41 years). In hereditary MTC the mean age of diagnosis was 27 +/- 13.9 years and was significantly lower than in sporadic one, where it was 45.7 +/- 14.3 years. The relationship between diagnosis, age and subtypes of hereditary MTC was assessed--no significant differences in examined subgroups were observed. The mean age of diagnosis in MEN 2A/FMTC and MEN 2A syndrome was 28-29 years, in MEN 2B - 21 years. The overall survival in sporadic MTC after 5 years was 97%, in hereditary MTC - 79%. Analysis performed after excluding suprarenal causes of death revealed no statistically significant differences in overall survival between both subtypes of MTC. 1. Hereditary MTC is still diagnosed too late, besides of DNA analysis. 2. In hereditary and sporadic MTC the prognosis is comparable.


Effect of adjunctive systemic azithromycin with periodontal surgery in the treatment of chronic periodontitis in smokers: a pilot study.
Along with conventional surgical therapy, systemic antibiotics may provide more effective treatment in smokers by targeting tissue-invasive bacteria. The aim of this randomized, placebo-controlled, double-masked clinical trial was to evaluate the adjunctive effects of systemic azithromycin (AZM) in combination with periodontal pocket reduction surgery in the treatment of chronic periodontitis in smokers. Thirty patients with a greater than one pack/day smoking habit and generalized moderate to severe chronic periodontitis were randomized to the test (surgery plus 3 days of AZM, 500 mg) or control group (surgery plus 3 days of placebo). Full-mouth probing depth (PD), clinical attachment level (CAL), bleeding on probing (BOP), gingival index (GI), plaque index, and wound healing indices (WHI) were assessed at baseline and at 2 weeks and 1, 3, and 6 months following surgical intervention. Plaque and gingival crevicular fluid were collected for trypsin-like enzyme activity (benzoyl-dl-arginine naphthylamine) and bone biomarker (crosslinked telopeptide of type I collagen [ICTP]) analyses, respectively, at baseline, 2 weeks, and 1, 3, and 6 months. Surgical treatment of moderate (PD = 4 to 6 mm) and deep (PD > 6 mm) pockets significantly improved clinical parameters of treated and untreated teeth (CAL gain, PD reduction, and reduction of BOP). The additional use of AZM did not enhance this improvement nor did it promote reduction of ICTP levels. Compared to the control group, the test group had significantly better WHI scores at 1 month, significantly less GI at 2 weeks, and sustained reductions of red-complex bacteria with trypsin-like enzyme activity at 3 months. For non-surgery teeth, only the test group showed significant gains in overall CAL compared to baseline. The findings of this pilot study demonstrated that in heavy smokers, adjunctive systemic AZM in combination with pocket reduction surgery did not significantly enhance PD reduction or CAL gain. However, the clinical value of adjunctive AZM may be appreciated by more rapid wound healing, less short-term gingival inflammation, and sustained reductions of periopathogenic bacteria. More expanded studies are recommended to better determine the clinical effects of adjunctive AZM in patients who smoke.


Anti-hepatoma activities of ethyl acetate extract from Ampelopsis sinica root.
Ampelopsis sinica root (ASR) is a known hepatoprotective folk traditional Chinese medicine. The anti‑hepatoma activity of ethyl acetate extract from A. sinica root (ASRE) in vitro and in vivo and its possible mechanism were explored. This study was designed to investigate cytotoxicity by MTT assay, induction of apoptosis via Hoechst 33258 staining, scanning electron microscopy and bivariate flow cytometric analysis (Annexin V-FITC/PI), inflammation and apoptosis related genes expression by RT-PCR and p53 protein expression by immunofluorescence assay in HepG2 cells. Then, the antitumor activity in vivo was detected by hepatoma H22 xenograft tumor in mice. The results showed that ASRE had powerful anti‑hepatoma activity in vitro without obvious toxicity on normal cells and could induce HepG2 cell apoptosis. The mechanism may be associated with downregulation of inflammatory cytokines including cyclooxygenase-2, 5-lipoxygenase and FLAP, increase of the ratio of bax/bcl-2, activation caspase-3 and inhibition of survivin, and increased expression of p53 protein. Furthermore, the HPLC assay showed the main compounds of ASRE were gallic acid, catechin and gallic acid ethyl ester. In animal experiments, ASR ethanol extract decreased the tumor weights of hepatoma H22 tumor-bearing mice. Therefore, ASR may be a potential therapeutic agent in the treatment of hepatocellular carcinoma.


How do we re-engage the pharmaceutical industry in research on serotonin and psychiatric disorders?
The Serotonin Club celebrated its silver jubilee in 2012 with a meeting in Montpellier, France. During the past 25 years, great advances have been made in our understanding of the pharmacology of serotonin receptors and the roles of this neurotransmitter in psychiatric disorders. Most of these advances have involved effective collaborations between academic and industrial scientists. In recent years, however, this picture has changed, as many of the major pharmaceutical companies have pulled out of in-house psychopharmacology research into the major psychiatric disorders, despite an increasing worldwide burden of these disorders and a clear need for improved treatment, particularly in terms of improved efficacy. This Viewpoint investigates the reasons for the decline in industrial involvement and makes proposals as to how future academic research on serotonin function in the brain might reawaken industry interest in serotonin-based research. Briefly, academic preclinical scientists need to alter their experimental approach to research into the psychiatric disorders. This will require a move from a single-target approach to understanding the complex neuronal pathways the cause diverse functional and behavioral outputs, using novel technological advances and the development of animal models with enhanced translational values. It is hoped that such an approach will reveal novel drug targets and thus re-engage the pharmaceutical industry in research that will result in improved human health and social well-being.


The Trouble with Tablets.
The biggest recommendation is to get your tablet screen up. This can be done by using a case or a tablet stand. Avoid using the tablet on your lap because posture follows vision. If you must use it on your lap, try placing it on a pillow to raise its height and reduce dangerous neck flexion. Whenever possible, set the tablet on a table rather than holding it with your hands because hands and forearms fatigue quickly. Consider finding an external keyboard that can be separated from the tablet screen. This way, you can set the tablet on a higher surface to elevate the screen and have a lower keyboard to eliminate some of the wrist extension when keying.


Social care interface in early-stage dementia: practitioners' perspectives on the links between formal and informal networks.
Although issues of complementarity and coordination between health care institutions have recently generated great interest, few scholars have studied the thorny interface between formal and informal service networks. This exploratory study investigates the interface between health care practitioners and caregivers of people with Alzheimer's disease living in the community. Using Pescosolido's multilevel network model and Martuccelli's sociology of the individual, the authors interviewed 20 practitioners drawn from seven urban groups and analyzed contacts between practitioners and caregivers at the beginning of the care trajectory. The quality of the practitioner-caregiver interface was determined by the players' profiles, internal structures, and external links. Heterogeneous professional groups and the establishment of contacts in the early stages of dementia seemed beneficial. The problematic interface between formal and informal service networks could be improved by a greater understanding of microsocial- and organizational-level interactions and implementing a social model of care.


Cognitive function and behavioural status in paediatric heart and heart-lung transplant recipients: the Harefield experience.
To assess the psychological impact of cardiac and cardiopulmonary transplantation on children. Retrospective cross sectional study. One British centre performing paediatric heart and heart-lung transplant operations, four cardiac units in London, three London schools, two London health centres, and the dental department of a London children's hospital. 65 children who had been given heart or heart-lung transplants and two reference groups of 52 children who had had other types of cardiac surgery and 45 healthy children. Development, cognition, and behaviour at home and at school as assessed by measures with proved validity and reliability. Developmental and cognitive measures indicated that children given transplants had significantly lower scores on several parameters, particularly in terms of development in children under 4 1/2 years of age. Performance on all tests, however, was within the normal range. There were no significant differences in behavioural ratings between the transplant and reference groups, though problem behaviour at home was more prevalent in the transplant group. Though cognitive development may be within the normal range, there are adverse psychological effects associated with cardiac and cardiopulmonary transplantation. These data indicate the need for a controlled prospective study in which children and their families are seen before and at regular intervals after transplantation. Interventions should be developed that are tailored to the particular needs of this very specialised group of paediatric patients and their families.


Recognizing syndromic hidradenitis suppurativa: a review of the literature.
Hidradenitis suppurativa (HS) is an inflammatory skin disease causing painful inflammation and suppuration. It may occur in rare syndromes: follicular occlusion, Bazex-Dupré-Christol, Down's, KID, PAPASH, PASS, PASH, and SAPHO syndromes, as well as Dowling-Degos disease. An overview of syndromic HS may inform the search for aetiological factors in HS. PubMed, Ovid and Web of Science were systematically searched using '(hidradenitis OR acne invers*) AND (syndrome OR KID OR PASS OR PAPA OR PASH OR SAPHO OR bazex-dupre OR 'dowling degos' OR triad OR tetrad)' and Cochrane Library using 'hidradenitis OR acne invers*'. A total of 82 articles were included in the final review. We summarize 134 cases collected from the 82 included articles. The syndromes are discussed, focusing on etiopathogenesis, clinical presentation and treatment. This study is based on case reports; therefore, conclusions may be subject to the selection bias. These syndromes are rare; however, it is important to recognize them, as treating them may require a different approach. Three subtypes of syndromic hidradenitis are suggested: syndromes with known genetic abnormalities, syndromes characterized by follicular plugging or structural defects, and syndromes with possible autoinflammatory pathogenesis. There was no universally effective treatment for syndromic HS, and treatment was individualized.


Total wrist arthroplasty. A clinical review.
Since 1974, a cementable, semi-constrained total wrist prosthesis has been implanted in patients with advanced destruction of the carpus usually secondary to rheumatoid arthritis. The single most common problem observed after surgery in the first one hundred cases was that of ulnar deformity. Since 1977, 25 total wrist arthroplasties have been performed in 22 patients (3 bilateral cases). A modification of the original prosthesis was employed in these. The newer designs allow for the precise reduplication of the instant center of motion of the normal wrist. Additionally, in the last six patients operated on, methylmethacrylate cement was not used to secure the metacarpal component because a secure press-fit was possible in each case. To date, postoperative results have disclosed no cases of radial ulnar imbalance. No cases of dislocation, infection, or loosening have occurred. No difference in the degree of postoperative pain relief has been observed between those cases in which cement was employed to secure the prosthetic components and those cases in which cement was used only to secure the radial component.


Clinician/patient connections in ethnoculturally nonconcordant encounters with political-asylum seekers: a comparison of physicians and nurses.
The article compares the ability of nurses and physicians to connect with patients in ethnoculturally nonconcordant clinical encounters with 41 randomly selected political-asylum seekers (PAS) residing at five Finnish reception centers in summer 2002. Doctors and nurses were equally unlikely to draw congruent assessments of the patient's past and present health condition, mixed use of biomedical/ethnocultural practices, adherence with medication and eat/drink instructions, (dis)satisfaction, and future confidence in recommended biomedical and ethnocultural approaches. Nurses were considerably more likely to hold views that were congruent with the patient's reported health care effectiveness in Finland. The findings suggest that doctors should request and place special weight on the insights of the principal attending nurse when assessing the potential contributions of personal, family, and host-society health care assets and inhibitors to a migrant patient's overall health plan. The results also suggest that culturally sensitive health care training offers specific advantages to nurses who attend to PAS.


Blood levels of neurofilament light are associated with disease progression in a mouse model of spinocerebellar ataxia type 3.
Increased neurofilament light (NfL; NEFL) protein in biofluids is reflective of neurodegeneration and has gained interest as a biomarker across neurodegenerative diseases. In spinocerebellar ataxia type 3 (SCA3), the most common dominantly inherited ataxia, patients exhibit progressive NfL increases in peripheral blood when becoming symptomatic, and NfL remains stably elevated throughout further disease course. However, progressive NfL changes are not yet validated in relevant preclinical SCA3 animal models, hindering its application as a biomarker during therapeutic development. We used ultra-sensitive single-molecule array (Simoa) to measure blood NfL over disease progression in YACQ84 mice, a model of SCA3, assessing relationships with measures of disease severity including age, CAG repeat size and magnetic resonance spectroscopy. YACQ84 mice exhibited plasma NfL increases that were concomitant with ataxia-related motor deficits as well as increased serum NfL, which correlated with previously established neurometabolite abnormalities, two relevant measures of disease in patients with SCA3. Our findings establish the progression of NfL increases in the preclinical YACQ84 mouse, further supporting the utility of blood NfL as a peripheral neurodegeneration biomarker and informing on coinciding timelines of different measures of SCA3 pathogenesis.


Lipopolysaccharide structures in Enterobacteriaceae, Pseudomonas aeruginosa, and Vibrio cholerae are immunologically related to Campylobacter spp.
To determine whether lipopolysaccharide (LPS) structures of Campylobacter species are immunologically related to those of 11 other gram-negative organisms, we immunoblotted from polyacrylamide gels the LPS of these strains with immune rabbit serum raised against six Campylobacter jejuni strains and two Campylobacter fetus strains. The LPS studied were from Salmonella minnesota wild type and Ra to Re mutants, Salmonella typhi, Escherichia coli, Yersinia enterocolitica, Vibrio cholerae, and Pseudomonas aeruginosa. None of the 11 LPS preparations was recognized by the eight antisera, but antisera to each of the Campylobacter strains recognized core determinants of some LPS preparations. Antiserum directed against the most serum-sensitive C. jejuni strain, 79-193, was the only antiserum sample that recognized core regions of the rough Salmonella mutants. In converse experiments, when LPS preparations from five Campylobacter strains were blotted with antiserum to Salmonella lipid A, recognition of core structures of each was shown; data from an enzyme-linked immunosorbent assay confirmed this result. In contrast, antiserum to Salmonella typhimurium Re LPS showed no reactivity. We conclude that LPS of Campylobacter strains share lipid A antigenic determinants with the core region of LPS of several other gram-negative organisms.


Ultrasensitive flow-based immunoassays using single-molecule counting.
Immunoassay (IA) technology has expanded the clinical utility of protein biomarkers, but demands for increased sensitivity, dynamic reporting ranges, and small sample volumes have limited the potential clinical usefulness of many biomarkers. We assessed the performance, including limits of detection (LODs) and the dynamic reporting range, of an IA-based technology, Erenna Immunoassay System, for a series of biomarkers, including cardiac troponin I (cTnI). Erenna IAs were used with 10 different and clinically important biomarkers to ascertain the LOD with various sample sizes (10 microL to 200 microL). The Erenna Immunoassay System generated LODs of 10-100 pg/L using 100 microL of sample. For cTnI, the LOD was 0.2 ng/L and a 10% CV was seen between 0.78 and 1.6 ng/L. The Erenna IA-based technology reproducibly measures protein biomarkers with detection limits of 10-100 pg/L, with a dynamic range of >4.5 logs in sample volumes of 50-150 microL.


Applicability of the International Affective Picture System in Chinese older adults: A validation study.
The International Affective Picture System (IAPS) is a standardized tool widely used to induce emotions in psychological studies. The present study examined the reliability and validity of the IAPS in Chinese older adults. A sample consisting of 126 Chinese older adults (86 females; aged 67.3 ± 4.96 years; 13.2 ± 2.75 years of education) was recruited to rate valence and arousal for 942 IAPS pictures. The results showed a satisfactory reliability and validity of the IAPS among these participants. The reliability was supported by high internal consistency and low inter-individual variance of participants' ratings; the validity was supported by high similarities (in rating scores and in the distribution of the pictures in the valence-arousal affective space) and small differences (in mean ratings) between Chinese and German older adults. In conclusion, the study adds supportive evidence to the cross-cultural validity of the IAPS in older adults, and provides a set of normative emotional ratings that could be adopted as a criterion in the selection of emotional pictures in future studies engaging Chinese older adults. Comparison of the IAPS ratings across cultures and ages is also discussed.


Molecular cloning of rat renin cDNA and its gene.
Renin (EC 126.96.36.199) is an aspartyl protease that cleaves its only known substrate, angiotensinogen, to release the vasopressor hormone angiotensin. We have isolated full-length cDNAs for renin from a rat kidney cDNA library. The cDNAs are complementary to a 1434-nucleotide rat kidney mRNA that encodes preprorenin, the 402-amino acid precursor of renin. This rat cDNA was used to isolate the complete copy of a renin gene from a rat genomic library, and a comparison of this genomic clone with rat genomic DNA showed that renin is a single-copy gene in the rat. Rat renin is 85% identical to one mouse renin isozyme (renin-1) and 81% identical to the second mouse renin isozyme (renin-2), suggesting that the duplication of the mouse renin genes is a more recent event than the speciation of rats and mice. Analyses of rat, human, and mouse renin sequences revealed that the potential to form two-chain renin is apparently peculiar to mouse renin, and the expression of a tenth exon (which results in a three-amino acid insertion) is observed only in the human renin gene.


Deep Bayesian-Assisted Keypoint Detection for Pose Estimation in Assembly Automation.
Pose estimation is crucial for automating assembly tasks, yet achieving sufficient accuracy for assembly automation remains challenging and part-specific. This paper presents a novel, streamlined approach to pose estimation that facilitates automation of assembly tasks. Our proposed method employs deep learning on a limited number of annotated images to identify a set of keypoints on the parts of interest. To compensate for network shortcomings and enhance accuracy we incorporated a Bayesian updating stage that leverages our detailed knowledge of the assembly part design. This Bayesian updating step refines the network output, significantly improving pose estimation accuracy. For this purpose, we utilized a subset of network-generated keypoint positions with higher quality as measurements, while for the remaining keypoints, the network outputs only serve as priors. The geometry data aid in constructing likelihood functions, which in turn result in enhanced posterior distributions of keypoint pixel positions. We then employed the maximum a posteriori (MAP) estimates of keypoint locations to obtain a final pose, allowing for an update to the nominal assembly trajectory. We evaluated our method on a 14-point snap-fit dash trim assembly for a Ford Mustang dashboard, demonstrating promising results. Our approach does not require tailoring to new applications, nor does it rely on extensive machine learning expertise or large amounts of training data. This makes our method a scalable and adaptable solution for the production floors.


Physical examination of the cervical spine and shoulder girdle in patients with shoulder complaints.
To look for differences in mobility in randomly selected patients without shoulder complaints depending on age, gender and left- or right-handedness; to investigate in the patient group whether specific differences exist, depending on the diagnosis made or the afflicted side; and to detect differences between the findings on the mobility examination of the cervical and cervicothoracic spine in patients with shoulder complaints compared with the healthy population. Comparative study. The physical examination findings after inclusion and after 26 wk of 101 shoulder patients were compared with those of 75 randomly selected patients without shoulder complaints and adjusted for age and gender. In the population of patients without shoulder complaints, the mobility in the cervical and upper thoracic spine was found to decrease with aging. Significant differences were found between the healthy and the afflicted population at the time of inclusion as well as after 26 wk, even though the majority of patients felt cured at that time. With all types of shoulder complaints, functional disorders in the shoulder girdle were found in which, at the time of inclusion, the emphasis was on pain and, after 26 wk, on restricted motion. Thus, functional disorders in the cervical spine, the higher thoracic spine and the adjoining ribs are not extrinsic causes of shoulder complaints, but an integral part of the intrinsic causes of shoulder complaints. Further study is needed to determine if this finding explains the high recurrence rate of shoulder complaints. It may be advisable to include the treatment of functional disorders of the shoulder girdle in treatment of patients with shoulder complaints.


Gastrostomy tube insertion into intestinal-cutaneous tract fistulas is a new technique to improve fistula control.
The management of gastrointestinal-cutaneous fistulas may be complicated by the difficulty in obtaining adequate control of the fistula tract. This study describes a new method to obtain better fistula control utilizing a semi-rigid stent in the form of a gastrostomy tube. Consecutive patients with intestinal-cutaneous fistulas of at least 3 weeks duration and treated by the new technique were analyzed. The technique involved the insertion of a guide wire into the fistula tract from the luminal side using an endoscope, snaring the wire with a Dormia basket inserted into the fistula tract from the cutaneous side and then exteriorized. The gastrostomy tube was then pulled with the guide wire from the lumen along the fistula tract and out through the skin. Five patients had had fistulas for a median duration of 42 (range 26-140) days before insertion of the gastrostomy tube. The gastrostomy tube was replaced with a smaller diameter tube in 4 of the patients (range 1-3 changes). The patients were discharged from the hospital at a median of 14 (range 12-23) days after the tube insertion but with the tube in situ. The median time from the insertion of the tube to its removal was 42 (range 32-108) days. Gastrostomy tube insertion using minimally invasive techniques may improve fistula control enabling patients to be discharged home sooner than otherwise and improve the rate of healing.


Effect of deoxycholic acid on Ca2+ movement, cell viability and apoptosis in human gastric cancer cells.
Deoxycholic acid (DOA) is one of the secondary bile acids used as a mild detergent for the isolation of membrane associated proteins. This study examined whether the secondary bile acid, DOA, altered Ca(2+) movement, cell viability and apoptosis in SCM1 human gastric cancer cells. The Ca(2+)-sensitive fluorescent dye fura-2 was used to measure [Ca(2+)]i. DOA-evoked [Ca(2+)]i rises concentration dependently. The response was reduced by removing extracellular Ca(2+). DOA-evoked Ca(2+) entry was inhibited by store-operated Ca(2+) channel inhibitors (nifedipine, econazole and SKF96365), the protein kinase C (PKC) activator phorbol 12-myristate 13 acetate (PMA) and the PKC inhibitor GF109203X. In Ca(2+)-free medium, treatment with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin (TG) abolished DOA-evoked [Ca(2+)]i rises. Conversely, treatment with DOA abolished TG-evoked [Ca(2+)]i rises. Inhibition of phospholipase C with U73122 abolished DOA-evoked [Ca(2+)]i rises. At 100-500 μM, DOA decreased cell viability, which was not changed by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). DOA between 100 and 300 μM also induced apoptosis. Collectively, in SCM1 cells, DOA-induced [Ca(2+)]i rises by evoking phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via store-operated Ca(2+) channels. DOA also caused Ca(2+)-independent apoptosis.


Vitamin D Signaling Pathways Confer the Susceptibility of Esophageal Squamous Cell Carcinoma in a Northern Chinese Population.
Experimental studies have determined the chemopreventive effects of vitamin D against the esophageal squamous cell carcinoma (ESCC); however, results from the epidemiological studies are not yet well established. The current study aimed to evaluate the associations between plasma vitamin D levels and variants on vitamin D metabolic-related genes with the risks for ESCC. A hospital-based case-control study was performed. Five hundred eighty-two ESCC patients and 569 controls were recruited in a Northern Chinese population. Common variants on vitamin D metabolism-related genes CYP24A1, DHCR7, GC, CYP27B1, and vitamin D receptor (VDR) and the plasma 25(OH)D level were determined. The unconditional logistic regression method was applied to determine the associations between the variants and vitamin D level and ESCC. Higher plasma 25(OH)D was associated with a reduced risk for ESCC, especially for rs2296241, rs11568820, and rs4646536. The variants rs2296241 on CYP24A1 and rs11568820 on VDR are significantly associated with ESCC cancer. Vitamin D signaling pathways may participate in the ESCC development. Further studies with larger sample size are warranted to confirm the results. Intervention studies are needed to determine whether vitamin D supplementation may reduce the ESCC risk in the Chinese population.


Psychoeducational Services for Children and Youth in the Orient: Current Status, Problems, and some Remedies.
Utilizing information obtained through an international survey and existing literature, patterns in the practices, research, and preparation of professionals who deliver psychoeducational services to children and youth in six Oriental countries (i.e., China, Hong Kong(1), Japan, the Philippines, Taiwan, and Thailand) are described. Services are provided by many professional groups. While few formal programs exist for the preparation of psychologists to work in schools, those providing such services have at least a bachelor's degree. Services commonly provided include assessment, vocational and educational guidance, counseling, parent education, and teaching. School psychological services generally are not governed by legislation or professional standards. Research tends to be applied and directed toward the construction and translations of tests and toward the needs of the mentally retarded, learning disabled, and behaviorally disordered. Ten major problems creating barriers for the delivery of psychoeducational services are identified. Three suggestions to help resolve these problems are offered.


Conservation of the carboxyl terminal epitope of myelin proteolipid protein in the tetrapods and lobe-finned fish.
Immunochemical analysis of the myelin proteolipid protein (PLP) has identified the carboxyl terminal amino acid phenylalanine 276 as the only PLP epitope conserved between the PLP components of rat and lungfish, species representing the phylogenetically most widely separated groups that synthesise typical CNS myelin. Immunoblotting using a rabbit antiserum raised against the carboxyl terminal sequence of rat PLP (residues 257-276) identified this epitope on the PLP components of both tetrapod (rat, chicken, lizard, and frog) and lobe-finned fish (coelacanth and lungfish) CNS myelin, including the DM-20 isoform of PLP, which is restricted to rat, chicken, and lizard CNS myelin. The conservation of the carboxyl terminus of PLP during evolution suggests this structure may play an important role in maintaining the organisation and function of PLP in the myelin membrane.


Breathing intensity and word use in individuals with COPD.
Shifts in natural or spontaneous language have been reported with psychological and physical changes, as well as shifts in predetermined words selected to describe dyspnea. Less evidence exists for natural language shifts and breathing intensity, particularly for everyday life and for negative emotion. Therefore, this study purpose was to describe the relationship between levels of everyday breathing intensity and the natural language word categories used in describing breathing, including a negative emotion category. A longitudinal descriptive research design and a convenience sample of 45 individuals were used. Natural language was analyzed as rates of word used based on three levels of breathing intensity. Non-parametric statistics were used to test differences between rates of word use and non-use on low, moderate, and high breathing intensity days, as well as correlations to subscales that measured negative emotion on the Bronchitis-Emphysema Symptom Checklist (BESC) and the Breathing Standard Index. Statistically significant associations were reported between level of breathing intensity and negative emotion, anger, cause, insight, time, and body words. Several significant correlations were reported with the psychosocial measures, including negative emotion and anxiety words with the BESC hopelessness/helplessness, time subscale, and negative emotion words with the inverse of typical breathing (i.e., atypical breathing). By showing significant associations between key word categories, in particular negative emotion and anger word use, levels of breathing intensity, and selected psychosocial measures, this study contributes to the body of knowledge about the influence of levels of breathing intensity on natural word use in everyday life.


Dietary practices of South Carolina adolescents and their parents.
To assess and identify correlates of adolescents' and parents' compliance with food guide pyramid recommendations (FGPR) and weight-control behaviors (WCB). Data were collected from a random sample of adolescents (2,021) and parents (1,231) and were analyzed using multiple and logistic regression. Only 7% eight graders, 4% eleventh graders, and 3% parents met all FGPR. The most significant predictors of FGPR and WCB include knowledge, attitudes, and eating concerns. Extreme dieters were less likely and moderate dieters were more likely to meet recommendations. Results have relevance for developing nutritional programs for adolescents.


[Effects of magnesium supply level on growth, nutrient element absorption and distribution, and quality of Panax quinquefolium].
This study aims to investigate the effects of different magnesium supply levels on the growth, nutrient absorption and distribution, and quality of Panax quinquefolium, and to determine the optimum content of exchangeable magnesium in soil. Three-year-old plants of P. quinquefolium were used in this study, and eight magnesium supply gradients(CK, Mg1-Mg7) were designed for indoor pot experiment(cultivation in soil). The plant growth indexes, nutrient element content in soil and plant, and root saponin content were determined at the end of the growth period. The correlation analysis of nutrient element content in aboveground and underground parts of P. quinquefolium showed significantly negative correlations of magnesium-calcium, magnesium-potassium, and magne-sium-manganese. With the increase in magnesium supply level, the biological absorption coefficient of magnesium increased, while that of total nitrogen, potassium, iron, and manganese decreased; the biological transfer coefficient of magnesium decreased, while that of nitrogen, phosphorus, calcium, iron, and manganese increased. The saponin content was analyzed by principal component analysis, which showed the comprehensive score in the order of Mg4(2.537), Mg2(1.001), Mg3(0.600), Mg1(0), Mg7(-0.765), CK(-0.825), Mg6(-0.922), and Mg5(-1.663). The partial least squares-path modeling(PLS-PM) showed that the correlation coefficients of exchangeable magnesium and pH with quality were-0.748 and-0.755, respectively, which were significant. Magnesium-calcium, magnesium-potassium, and magnesium-manganese showed antagonism in the nutritional physiology of P. quinquefolium. Excessive application of magnesium can lead to the imbalance of nutrient elements in P. quinquefolium. The content of exchangeable magnesium in soil suitable for the quality formation of P. quinquefolium was 193.34-293.34 mg·kg~(-1). In addition to exchangeable magnesium, pH was also important to the quality formation of P. quinquefolium. Therefore, exchangeable magnesium and pH could be regarded as monitoring factors for the quality formation of P. quinquefolium.


On optically stimulated luminescence properties of household salt as a retrospective dosemeter.
Thermoluminescence (TL) and optically stimulated luminescence (OSL) in the UV (270-370 nm) spectral region have been investigated for five types of table salt (NaCl) available in Romanian supermarkets with a view to applying them in retrospective dosimetry. The salt samples gave bright TL signals with two main peaks at ∼100°C and at 300 or 260°C, depending on the origin of the salt and bright OSL signals under continuous stimulation with blue light. The OSL signal (stimulated at 100°C after a pre-heat of 10 s at 150°C) was used for investigations in a standard multiple aliquot procedure. The dose-response was found to be linear in the dose range investigated (up to ∼100 mGy) and the lower limit of detection for the samples varied from ∼0.01 to 14 mGy. These characteristics, along with the widespread abundance and low cost of household salt, confirm its potential as a retrospective dosemeter.


ZnVSe2O7 and Cd6V2Se5O21: new d10 transition-metal selenites with V(IV) or V(V) cations.
Two new metal selenites with a combination of vanadium(IV) or vanadium(V) cations, namely, ZnVSe 2O 7 and Cd 6V 2Se 5O 21, have been synthesized by hydrothermal and high-temperature solid-state reactions, respectively. The structure of ZnVSe 2O 7 features a 3D network of vanadium(IV) selenite with 1D tunnels occupied by zinc(II) ions. The 3D network of vanadium(IV) selenite is formed by corner-sharing V (IV)O 6 octahedral chains bridged by selenite groups. In Cd 6V 2Se 5O 21, the interconnection of cadmium(II) ions by bridging and chelating selenite groups led to a 3D framework with large tunnels along the b axis, and the 1D chains of corner-sharing V (V)O 4 tetrahedra are inserted in the above large tunnels and are bonded to the cadmium selenite framework via Cd-O-V bridges. Both compounds exhibit broad emission bands in the blue-light region. Results of magnetic property measurements show there is significant antiferromagnetic interaction between V (4+) centers in ZnVSe 2O 7. The electronic structure calculations for both compounds have been also performed.


Actual fusion efficiency in the lipid mixing assay - Comparison between nanodiscs and liposomes.
Lipid exchange occurs between membranes during fusion or active lipid transfer. These processes are necessary in vivo for the homeostasis of the cell at the level of the membranes, the organelles and the cell itself. They are also used by the cell to interact with the surrounding medium. Several assays have been developed to characterize in vitro these processes on model systems. The most common one, relying on fluorescence dequenching, measures lipid mixing between small membranes such as liposomes or nanodiscs in bulk. Usually, relative comparisons of the rate of lipid exchange are made between measurements performed in parallel. Here, we establish a quantitative standardization of this assay to avoid any bias resulting from the temperatures, the chosen fluorescent lipid fractions and from the various detergents used to normalize the measurements. We used this standardization to quantitatively compare the efficiency of SNARE-induced fusion in liposome-liposome and liposome-nanodisc configurations having similar collision frequency. We found that the initial yield of fusion is comparable in both cases, 1 per 2-3 million collisions in spite of a much larger dequenching signal with nanodiscs. Also, the long-term actual fusion rate is slightly lower with nanodiscs than in the liposome-liposome assay.


Contrasting genes conferring short- and long-term biofilm adaptation in Listeria.
Listeria monocytogenes is an opportunistic food-borne bacterium that is capable of infecting humans with high rates of hospitalization and mortality. Natural populations are genotypically and phenotypically variable, with some lineages being responsible for most human infections. The success of L. monocytogenes is linked to its capacity to persist on food and in the environment. Biofilms are an important feature that allow these bacteria to persist and infect humans, so understanding the genetic basis of biofilm formation is key to understanding transmission. We sought to investigate the biofilm-forming ability of L. monocytogenes by identifying genetic variation that underlies biofilm formation in natural populations using genome-wide association studies (GWAS). Changes in gene expression of specific strains during biofilm formation were then investigated using RNA sequencing (RNA-seq). Genetic variation associated with enhanced biofilm formation was identified in 273 genes by GWAS and differential expression in 220 genes by RNA-seq. Statistical analyses show that the number of overlapping genes flagged by either type of experiment is less than expected by random sampling. This novel finding is consistent with an evolutionary scenario where rapid adaptation is driven by variation in gene expression of pioneer genes, and this is followed by slower adaptation driven by nucleotide changes within the core genome.


Distribution of Cytokeratin 17 in the Parenchymal Elements of Rat's Submandibular Glands Subjected to Fractionated Radiotherapy.
The aim of this research was to study the intensity of cytokeratin 17 (CK17) in the parenchymal elements of rat's submandibular salivary glands subjected to fractionated radiotherapy regimen that used for treatment of head and neck malignancy. Twenty male albino rats were divided into two equal groups (normal and irradiated). The irradiated group received a radiation dose of 5 Grays daily for 5 days using therapeutic X-ray beam. Six months later, submandibular gland was dissected out and prepared for both histological and immunohistochemical studies. Submandibular gland of irradiated group showed two different types of histological alterations. The first alteration showed severe gland atrophy replaced by either fibrous or fatty tissues. In some sections, the gland exhibited proliferating activity in the form of profuse amounts of mitotic figures. Immunohistochemical examination of control glands displayed a mild cytoplasmic expression of CK17 of duct cells as well as serous acini. The staining pattern was either diffused or concentrated at the basal part of the cell with negative expression at its apical part. Expression of CK17 in submandibular gland of irradiated group displayed a highly significant differences (P < 0.001) in both intercalated and striated ducts. Many serous acini displayed a highly significant differences (P < 0.001) whereas, mucous acini were negatively stained. The intensity and diffusion of CK17 expression in our results foretell the pathological effect of radiotherapy on the intermediate filaments of salivary gland parenchyma that interfered with production and/or secretion of saliva leading to xerostomia.


Theory of critical enhancement of photorefractive beam coupling.
We develop the idea of critical enhancement of the photorefractive response near the threshold of parametric excitation of space-charge waves (the spatial subharmonics) taking into account the vectorial character of beam coupling and a fairly strong broadening of the nonlinear resonance owing to light absorption. The results of our calculations are a description of the measurable characteristics of critical enhancement and optimization of the experimental conditions for detection of anomalously high amplification gain factors in cubic Bi(12)SiO(20) crystals.


Acquired forms of the Brugada syndrome.
The Brugada syndrome is characterized by ST-segment elevation in the right precordial leads (V1 through V3) and an episode of ventricular fibrillation in the absence of structural heart disease. SCN5A, the gene encoding the alpha subunit of the sodium channel, is the only gene thus far linked to the Brugada syndrome but is identified in only 18% to 30% of patients with clinically diagnosed Brugada syndrome. On the other hand, experimental studies have suggested that an intrinsically prominent transient outward current-mediated action potential (AP) notch and a subsequent loss of the AP dome in the epicardium but not in the endocardium of the right ventricular outflow tract give rise to a transmural voltage gradient, resulting in ST-segment elevation and phase 2 reentry-induced ventricular fibrillation. Therefore, any intervention that increases outward currents (eg, transient outward current, adenosine triphosphate-sensitive potassium current, delayed modifier potassium current) or decreases inward currents (eg, L-type calcium current, fast sodium current) at the end of phase 1 of the AP can accentuate or unmask ST-segment elevation, similar to that found in the Brugada syndrome, thus producing acquired forms of the Brugada syndrome. In this review, several drugs in addition to sodium-channel blockers and conditions that induce transient ST-segment elevation such as that in the Brugada syndrome, developing acquired forms of the Brugada syndrome, are discussed.


TREM2 is required for microglial instruction of astrocytic synaptic engulfment in neurodevelopment.
Variants in the microglial receptor TREM2 confer risk for multiple neurodegenerative diseases. However, it remains unknown how this receptor functions on microglia to modulate these diverse neuropathologies. To understand the role of TREM2 on microglia more generally, we investigated changes in microglial function in Trem2-/- mice. We found that loss of TREM2 impairs normal neurodevelopment, resulting in reduced synapse number across the cortex and hippocampus in 1-month-old mice. This reduction in synapse number was not due directly to alterations in interactions between microglia and synapses. Rather, TREM2 was required for microglia to limit synaptic engulfment by astrocytes during development. While these changes were largely normalized later in adulthood, high fat diet administration was sufficient to reinitiate TREM2-dependent modulation of synapse loss. Together, this identifies a novel role for microglia in instructing synaptic pruning by astrocytes to broadly regulate appropriate synaptic refinement, and suggests novel candidate mechanisms for how TREM2 and microglia could influence synaptic loss in brain injury and disease.


Failure to pop out: Feature singletons do not capture attention under low signal-to-noise ratio conditions.
Pop-out search implies that the target is always the first item selected, no matter how many distractors are presented. However, increasing evidence indicates that search is not entirely independent of display density even for pop-out targets: search is slower with sparse (few distractors) than with dense displays (many distractors). Despite its significance, the cause of this anomaly remains unclear. We investigated several mechanisms that could slow down search for pop-out targets. Consistent with the assumption that pop-out targets frequently fail to pop out in sparse displays, we observed greater variability of search duration for sparse displays relative to dense. Computational modeling of the response time distributions also supported the view that pop-out targets fail to pop out in sparse displays. Our findings strongly question the classical assumption that early processing of pop-out targets is independent of the distractors. Rather, the density of distractors critically influences whether or not a stimulus pops out. These results call for new, more reliable measures of pop-out search and potentially a reinterpretation of studies that used relatively sparse displays. (PsycINFO Database Record


IL-1Rahigh-IL-4low-IL-13low: A Novel Plasma Cytokine Signature Associated with Olfactory Dysfunction in Older US Adults.
Inflammation has been implicated in physical frailty, but its role in sensory impairment is unclear. Given that olfactory impairment predicts dementia and mortality, determining the role of the immune system in olfactory dysfunction would provide insights mechanisms of neurosensory decline. We analyzed data from the National Social Life, Health and Aging Project, a representative sample of home-dwelling older US adults. Plasma levels of 18 cytokines were measured using standard protocols (Luminex xMAP). Olfactory function was assessed with validated tools (n-butanol sensitivity and odor identification, each via Sniffin' Sticks). We tested the association between cytokine profiles and olfactory function using multivariate ordinal logistic regression, adjusting for age, gender, race/ethnicity, education level, cognitive function, smoking status, and comorbidity. Older adults with the IL-1Rahigh-IL-4low-IL-13low cytokine profile had worse n-butanol odor sensitivity (odds ratio [OR] = 1.61, 95% confidence interval [CI] 1.19-2.17) and worse odor identification (OR = 1.42, 95% CI 1.11-1.80). Proinflammatory, Th1, or Th2 cytokine profiles were not associated with olfactory function. Moreover, accounting for physical frailty did not alter the main findings. In conclusion, we identified a plasma cytokine signature-IL-1Rahigh-IL-4low-IL-13low-that is associated with olfactory dysfunction in older US adults. These data implicate systemic inflammation in age-related olfactory dysfunction and support a role for immune mechanisms in this process, a concept that warrants additional scrutiny.


Corpuscular bodies in the palate of the rat. 2. Innervation and central projection.
The source of innervation of the corpuscular bodies in the palate and the central projections of the afferent fibres of the entire palate was studied in rats by transganglionic transport of horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP) and with substance P (SP) immunohistochemistry. WGA-HRP injected into the incisal papilla was taken up by the nerve fibres that terminated in the corpuscles. Retrogradely labelled neurons were observed in the trigeminal ganglion as well as anterogradely labelled terminals in the dorsolateral part of the spinal trigeminal nucleus and in the lateral part of the nucleus of the solitary tract. No labelling could be found in the geniculate ganglion, the facial nerve and the hypoglossal nucleus. Following WGA-HRP injection in the intermolar area and in the soft palate, labelling was only restricted to the trigeminal ganglion. The lamina propria of the entire palate and the corpuscle-enriched area of the incisal papilla and the soft palate were richly innervated by SP-containing fibres. Numerous SP-containing fibres were also observed in the nerve plexus at the base of the corpuscle. In addition, SP-positive neurons were identified in the trigeminal ganglion and SP-labelled terminals in the sensory trigeminal nuclear complex and in the solitary tract nucleus. On the basis of our morphological observations we conclude that the palatal corpuscular bodies are involved in taste perception which is of trigeminal origin.


Isolation and culture of microvascular endothelial cells from gliomas of different WHO grades.
Gliomas are the most common intrinsic brain tumors. The degree of vascularization corresponds to malignancy and is related to prognosis. In order to retrieve information about tumor behavior in situ, the use of primary tissue material for experiments is advantageous. With increasing evidence for the importance of microenvironment and vascularization in tumor biology, we concentrated on the isolation of endothelial cells (EC) from primary tumor material to investigate the role of endothelium within tumor tissue. We developed a method for isolation and purification of tumor-derived endothelial cells. EC were isolated and cultivated from normal brain using tissue digestion and Percoll density gradient centrifugation resulting in a <95% of EC culture. For isolation of EC from gliomas of different malignancy grades a combination of tissue digestion, Percoll gradient centrifugation and magnetic bead sorting by anti-CD31, -VE-Cadherin and -CD 105 was employed. This approach provided a purity of <98%. Cells were classified and characterized by testing expression of CD105, CD31, VE-Cadherin, vWF, UEA-1 and measuring DiI-Ac-LDL-uptake. To exclude contamination, staining and negative selection with anti-SMA, -GFAP, and -CD68 was performed. Tumors were histopathologically diagnosed according to WHO classification. We isolated EC from normal brain (NBEC, n = 11), low-grade gliomas WHO II (LGEC, n = 22), and high-grade gliomas WHO III & IV (HGEC, n = 11). There were no clear differences in EC morphology between the different tumor grades. However, a significantly higher proliferation rate of HGEC compared to LGEC was observed as well as distinctive antigen expression. Already in early passages isolated EC showed a rapid change in antigen expression indicating a phenotypic shift under culture conditions. We could establish a protocol for reliable and reproducible isolation and culture of EC from gliomas with different WHO grading. In first phenotypical and functional analyses, NBEC, LGEC and HGEC show remarkable differences. EC from all tumors could be grown in culture. However, passage related changes of EC phenotype demand very early passages to work with.


Not just little adults: palliative care physician attitudes toward pediatric patients.
Palliative care physicians are increasingly being asked to provide end-of-life (EOL) care for children. Yet very little is known about physicians' level of comfort and willingness to do so. This study assessed the attitudes of palliative care physicians toward providing care for pediatric patients and to describe the supports they desire in order to do so. An online questionnaire was e-mailed to all physicians in the Division of Palliative Care at the University of Toronto. The questionnaire explored perceptions, attitudes, and level of comfort caring for pediatric patients. Results are reported using frequencies, ratios, and other descriptive analyses. Forty-four physicians of the 74 (59%) surveyed responded. On average, physicians cared for fewer than one child per each year of practice. Although the majority of respondents perceived their pediatric training to be inadequate, 70% were willing to provide care to children. Respondents felt at ease applying their knowledge and skills in some aspects of pediatric care (e.g., principles of pain and symptom management, communication about EOL issues) but less so in others (e.g., medication dosing, ethical issues unique to pediatrics). All respondents welcomed opportunities for additional training, but a third felt it was not essential. In particular, the most frequently expressed need was for mentorship by pediatric palliative care specialists. Palliative physicians tend to be willing to care for children, but perceive their level of training to be insufficient. Although additional training is endorsed, physicians favored real-time support and mentorship from a pediatric expert.


Variations in bile tolerance among Lactococcus lactis strains derived from different sources.
Lactococcus lactis subsp. lactis has been isolated from the intestines of marine fish and is a candidate probiotic for aquaculture. In order to use the bacterium as a probiotic, properties such as bile tolerance need to be assessed. Here, we compared bile tolerance in L. lactis strains derived from different sources. Three L. lactis subsp. lactis strains from marine fish (MFL), freshwater fish (FFL), and cheese starter (CSL) were used along with an Lactococcus lactis subsp. cremoris strain from cheese starter (CSC). The four strains were grown under various culture conditions: deMan-Rogosa-Sharpe (MRS) broth containing bile salts/acids, MRS agar containing oxgall, and phosphate-buffered saline (PBS) containing fish bile. Survival/growth of the strains in the presence of sodium cholate and sodium deoxycholate varied in the order MFL, CSL > CSC > FFL; in the presence of sodium taurocholate, the order was MFL > CSL > CSC > FFL. In liquid media containing various concentrations of oxgall, survival of the strains was observed in the order MFL > CSL > FFL and CSC. The survival of MFL was not affected by bile collected from the goldfish (Carassius auratus subsp. auratus) or the puffer fish (Takifugu niphobles), although the other strains showed significant inhibition of growth. It is a novel and beneficial finding that MFL has the highest resistance to bile acid.


Relationship among risk factors of atherosclerosis, leukocyte count, and soluble intercellular adhesion molecule-1.
We investigated the effects of risk factors of atherosclerosis on soluble intercellular adhesion molecule-1 (sICAM-1) concentration and leukocyte count by using multivariate statistical analysis. The subjects were 90 people who were hospitalized for a complete check-up examination. Eight risk factors were selected as candidates for predictor variables: age, body mass index (BMI), uric acid, glycohemoglobin A1c, number of cigarettes smoked daily, total cholesterol, triglyceride, and high density lipoprotein (HDL)-cholesterol. The predictor variables were selected using a stepwise method, [criterion variable - predictor variable (standardized regression coefficient)]; sICAM1 - age (0.1859), number of cigarettes (0.2558), triglyceride (0.2447); leukocytes - number of cigarettes (0.2827), triglyceride (0.2526), HDL-cholesterol (-0.2800); stab leukocytes - number of cigarettes (0.2460); segmented leukocytes - glycohemoglobin A1c (0.1495), number of cigarettes (0.2716), HDL-cholesterol (-0.3254); lymphocytes - BMI (0.2639), number of cigarettes (0.1495), triglyceride (0.3520); monocytes - glycohemoglobin A1c (0.2617). These findings indicated that the risk factors of atherosclerosis may influence sICAM1 concentration and leukocyte count.


The Streptomyces galP1 promoter has a novel RNA polymerase recognition sequence and is transcribed by a new form of RNA polymerase in vitro.
We report the identification of DNA sequences that determine the activity of the Streptomyces galP1 promoter and a new form of RNA polymerase holoenzyme that recognizes these sequences in vitro. Base substitutions were introduced throughout the galP1 promoter region, and bases at positions -34, -36, and -11 with respect to the transcription start site were shown to be required for promoter function. These bases correspond in their positions to regions known to be important for RNA polymerase binding in several classes of eubacterial promoters, but the sequences themselves are not similar to those previously described. The -35 region of the galP1 promoter consists of six G residues, and base changes in this G hexamer had a dramatic effect on promoter activity. By using galP1-containing DNA template, a new RNA polymerase activity was purified from Streptomyces. Holoenzyme reconstitution experiments identified a new sigma factor that directs galP1 transcription in vitro. DNase I protection experiments identified a binding site for this new holoenzyme immediately upstream of the galP1 transcription start site.


Prevalence of multiple drug-resistant Plasmodium falciparum malaria cases in Northeast India.
Two numbers of Plasmodium falciparum field isolates from Gossingpara, Runikhata area in Chirang district of Assam had shown multiple mutations in Pfcrt-dhfr-dhps gene (up to seven mutations: One mutation in Pfcrt gene, three mutations in Pfdhfr gene and three mutations in Pfdhps gene). Similarly, two cases in Bat camp, Miao area under Changlang district of Arunachal Pradesh had shown a total of eight mutations, of which one mutation in Pfcrt gene, three mutations in Pfdhfr gene, three mutations in Pfdhps gene and one mutation in PfATPase6 gene. One case in 3 Miles, Miao area of Changlang district has shown mutations in Pfcrt(one mutation), Pfdhfr(four mutations) and Pfdhps(three mutations) gene. These results indicated that there is an existence of multiple mutant P. falciparum malaria cases in northeastern region of India.


[Comparative analysis of genetic structure of 2 species of marine fish Dicentrarchus labrax and Dicentrarchus punctatus].
We present here the genetic structure existing among five samples of the spotted sea bass Dicentrarchus punctatus, and we compare it to what prevails in the common sea bass D. labrax, a congeneric species sampled on almost the same geographical range. A genetic distance tree inferred from the polymorphism at six microsatellite loci shows a distinct pattern for the two species. D. labrax samples appears to be genetically more homogeneous with a global Fst of 3% as compared to the 10% observed at D. punctatus, indicating a lesser level of gene flow in the latter species. While appearing more differentiated, D. punctatus presents no clear geographical organisation of its genetic variability in opposition to D. labrax samples. This allows us to propose this pair of closely relative species as a good candidate for the study by comparative analysis of the biological and/or historical factors affecting genetic differentiation in marine environment.


Changes in bacterial community metabolism and composition during the degradation of dissolved organic matter from the jellyfish Aurelia aurita in a Mediterranean coastal lagoon.
Spatial increases and temporal shifts in outbreaks of gelatinous plankton have been observed over the past several decades in many estuarine and coastal ecosystems. The effects of these blooms on marine ecosystem functioning and particularly on the dynamics of the heterotrophic bacteria are still unclear. The response of the bacterial community from a Mediterranean coastal lagoon to the addition of dissolved organic matter (DOM) from the jellyfish Aurelia aurita, corresponding to an enrichment of dissolved organic carbon (DOC) by 1.4, was assessed for 22 days in microcosms (8 l). The high bioavailability of this material led to (i) a rapid mineralization of the DOC and dissolved organic nitrogen from the jellyfish and (ii) the accumulation of high concentrations of ammonium and orthophosphate in the water column. DOM from jellyfish greatly stimulated heterotrophic prokaryotic production and respiration rates during the first 2 days; then, these activities showed a continuous decay until reaching those measured in the control microcosms (lagoon water only) at the end of the experiment. Bacterial growth efficiency remained below 20%, indicating that most of the DOM was respired and a minor part was channeled to biomass production. Changes in bacterial diversity were assessed by tag pyrosequencing of partial bacterial 16S rRNA genes, DNA fingerprints, and a cultivation approach. While bacterial diversity in control microcosms showed little changes during the experiment, the addition of DOM from the jellyfish induced a rapid growth of Pseudoalteromonas and Vibrio species that were isolated. After 9 days, the bacterial community was dominated by Bacteroidetes, which appeared more adapted to metabolize high-molecular-weight DOM. At the end of the experiment, the bacterial community shifted toward a higher proportion of Alphaproteobacteria. Resilience of the bacterial community after the addition of DOM from the jellyfish was higher for metabolic functions than diversity, suggesting that jellyfish blooms can induce durable changes in the bacterial community structure in coastal lagoons.


MDM2 Antagonist Nutlin-3 Stimulates Global DNA Hydroxymethylation by Enhancing p53-TET1 Signaling Axis.
DNA hydroxymethylation is involved in many biological processes, including nuclear reprogramming, embryonic development, and tumor suppression. In this study, we report that an anticancer agent, nutlin-3, selectively stimulates global DNA hydroxymethylation in TP53 wild-type cancer cells as manifested by the elevation of 5-hydroxymethylcytosine (5hmC) in genomic DNA. In contrast, nutlin 3 fails to enhance DNA hydroxymethylation in TP53-mutated cancer cells. Consistently, nutlin-3 as a MDM2 antagonist only activates wild-type but not mutated TP53. Furthermore, nutlin-3 does not alter the expression of TET1 but slightly reduces the expression of TET2 and TET3 proteins. These TET family proteins are responsible for converting 5-methylcytosine (5mC) to 5hmC. Interestingly, TET1 knockdown could significantly block the nutlin-3-induced DNA hydroxymethylation as well as TP53 and P21 activation. Immunoprecipitation analysis supports that p53 strongly interacts with TET1 proteins. These results suggest that nutlin-3 activates TP53 and promotes p53-TET1 interaction. As positive feedback, the p53-TET1 interaction further enhances p53 activation and promotes apoptosis. Collectively, we demonstrate that nutlin-3 stimulates DNA hydroxymethylation and apoptosis via a positive feedback mechanism.


A Microwell-Printing Fabrication Strategy for the On-Chip Templated Biosynthesis of Protein Microarrays for Surface Plasmon Resonance Imaging.
A two-step templated, ribosomal biosynthesis/printing method for the fabrication of protein microarrays for surface plasmon resonance imaging (SPRI) measurements is demonstrated. In the first step, a sixteen component microarray of proteins is created in microwells by cell free on chip protein synthesis; each microwell contains both an in vitro transcription and translation (IVTT) solution and 350 femtomoles of a specific DNA template sequence that together are used to create approximately 40 picomoles of a specific hexahistidine-tagged protein. In the second step, the protein microwell array is used to contact print one or more protein microarrays onto nitrilotriacetic acid (NTA)-functionalized gold thin film SPRI chips for real-time SPRI surface bioaffinity adsorption measurements. Even though each microwell array element only contains approximately 40 picomoles of protein, the concentration is sufficiently high for the efficient bioaffinity adsorption and capture of the approximately 100 femtomoles of hexahistidine-tagged protein required to create each SPRI microarray element. As a first example, the protein biosynthesis process is verified with fluorescence imaging measurements of a microwell array containing His-tagged green fluorescent protein (GFP), yellow fluorescent protein (YFP) and mCherry (RFP), and then the fidelity of SPRI chips printed from this protein microwell array is ascertained by measuring the real-time adsorption of various antibodies specific to these three structurally related proteins. This greatly simplified two-step synthesis/printing fabrication methodology eliminates most of the handling, purification and processing steps normally required in the synthesis of multiple protein probes, and enables the rapid fabrication of SPRI protein microarrays from DNA templates for the study of protein-protein bioaffinity interactions.


Changes in peripheral blood Th1 and Th2 cells in rat liver transplantation under different immune statuses.
In this study, early expressions of peripheral blood Th1 and Th2 cells were documented following rat liver transplantation and related to immune status. Rats were divided into 3 groups: group A (control): syngeneic transplantation (Brown Norway (BN) → BN); group B: allogeneic transplantation + cyclosporine A (CsA); group C: allogeneic transplantation (Lewis → BN). Flow cytometry was used to analyze peripheral blood CD4(+)CD45RC percentage on days 1, 3, 5, 7, and 14 following transplantation, and were compared to graft rejection pathological grades and receptor survival times. The average survival of groups A and B exceeded 100 days, which was significantly longer than that of group C (3.56 ± 34.3 days). With the exception of the first day, rejection grades were significantly higher in groups C and B compared to group A, and group C rejection grades were significantly higher than those of group B. Three days after transplantation, the CD4(+)CD45RC(+) to CD4(+)CD45RC(-) ratio of group C was significantly higher than that of groups A and B. In group B, the CD4(+)CD45RC(+) to CD4(+)CD45RC(-) ratio was negatively correlated to the rejection grade (r = -0.565, P < 0.01), whereas this relationship was positive in group C (r = 0.745, P < 0.01). In conclusion, peripheral blood Th1 was highly expressed during rejection in rat liver grafts. Peripheral blood Th2 tended to increase early under rejection inhibition with CsA, and its high expression level may correlate with long-term acceptance or tolerance of transplanted livers.


Berberine remodels adipose tissue to attenuate metabolic disorders by activating sirtuin 3.
Adipose tissue remodelling is considered a critical pathophysiological hallmark of obesity and related metabolic diseases. Berberine (BBR), a natural isoquinoline alkaloid, has potent anti-hyperlipidaemic and anti-hyperglycaemic effects. This study aimed to explore the role of BBR in modulating adipose tissue remodelling and the underlying mechanisms. BBR protected high fat diet (HFD)-fed mice against adiposity, insulin resistance and hyperlipidemia. BBR alleviated adipose tissue inflammation and fibrosis by inhibiting macrophage infiltration, pro-inflammatory macrophage polarization and the abnormal deposition of extracellular matrix, and the effect was mediated by BBR directly binding and activating the deacetylase Sirtuin 3 (SIRT3) and suppressing the activation of the mitogen-activated protein kinases and nuclear factor-κB signalling pathways. Furthermore, BBR decreased microRNA-155-5p secretion by macrophages, which in turn ameliorated liver injury. Moreover, BBR mitigated inflammatory responses in both LPS-stimulated macrophages and TNF-α-treated adipocytes and suppressed macrophage migration towards adipocytes by activating SIRT3. Collectively, this study revealed that BBR improved adipose tissue remodelling, and subsequently inhibited the secretion of microRNA-155-5p by macrophages, which alleviated adiposity, insulin resistance and liver injury in obese mice. The modulation of adipose tissue remodelling by activating SIRT3 could contribute to the anti-hyperlipidemic and anti-hyperglycemic effects of BBR.