Black African migrants: the barriers with accessing and utilizing health promotion services in the UK.
The inequalities for different ethnicities and social classes in accessing health services is well documented, but although a number of recent policy developments have aimed to tackle health inequalities, very little is known about the experiences of Black African migrant communities in accessing health promotion information and services. The aim of the study were to examine the experiences of Black African migrant families in accessing health promotion services. A convenience sample of 90 Black African migrants in the north of England participated in the study. Quantitative and qualitative data were collected utilizing a self-administered questionnaire in Arabic, English, French and Swahili. The findings were analysed using descriptive analysis and a variation of the constant comparative method. Themes and categories were identified across transcripts and appropriate quotations have been used to illustrate themes. Major findings that emerged from the analysis showed that participants were concerned about their insufficient ability to communicate, which appeared to undermine their capacity to access health promotion services. Lack of literacy and proficiency in English was perceived as an underlying problem in seeking health promotion information and support. The findings have a number of implications for health promotion practitioners; in particular, the need to work collaboratively with migrant groups in order to identify and develop appropriate cultural sensitive communication strategies. The study concludes by suggesting the need to explore further the communication needs of migrant families and the implications for the take-up of health promotion services.


Mortality Rate and Antibiotic Resistance in Complicated Diverticulitis: Report of 272 Consecutive Patients Worldwide: A Prospective Cohort Study.
Background: Complicated diverticulitis (CD) is a common problem for surgeons. In treating it, as a general principle, every verified source of infection should be controlled. Supplementary antimicrobial management involves a delicate balance of optimizing empirical therapy while reducing unnecessary antibiotic use. The necessity to know the most frequent infecting pathogens and their spectra of resistance is becoming pivotal. The aim of this study was to determine the microbiologic profile of complicated intra-abdominal infections (IAIs) secondary to CD, to analyze the role of empirical antimicrobial therapy, and to describe the clinical aspects of CD worldwide. Material and Methods: The study derives from two multicenter prospective observational studies: CIAO (Complicated Intra-Abdominal infection Observational study) and CIAOW (Complicated Intra-Abdominal infection Observational World" study). The aim of the study was to analyze the intra-abdominal bacteriology in complicated diverticulitis and its relation to the clinical outcome. Results: The 272 patients had a mean/median age of 66.3 ± 14.9 (standard deviation; SD) and 69 (range 18-99). Patients >70 years old totaled 122 (44.9%). Conditions at admission were sepsis in 113 patients (41.5%) and severe sepsis and septic shock in 37 (13.6%) and 21 (7.7%), respectively; and localized peritonitis in 148 patients (54.4%), whereas in 124 (45.6%), the condition was generalized. Some 94 patients (34.6%) experienced a delay in initial intervention (>24 h). The mean and median duration of antimicrobial therapy were 12.3 ± 8.7 (standard deviation) and 10 (range 1-59) days. A total of 162 patients (59.6%) obtained adequate empirical antibiotic therapy, and 96 patients (35.3%) were admitted to the intensive care unit (ICU). The 30-day mortality rate was 12.1%. A total of 311 bacteria were isolated: 136 (43.7%) gram-negative, 76 (24.4%) gram-positive, 22 (7%) fungi, and 77 (24.7%) anaerobes. Of the 363 bacteria isolated, 22 (7%) were drug resistant. Four of these infections (22.2%) were health-care-associated and 18 (5.7%) community-acquired. By univariable analysis, the only statistically significant factor associated with resistant bacteria was inadequacy of the empirical antimicrobial therapy (p = 0.004). The factors associated with death were delay in initial intervention (p = 0.006) and ICU admission because of severe sepsis on admission (p = 0.004). Conclusion: Early source control is mandatory to reduce the mortality rate in complicated diverticulitis. Effective empirical antimicrobial agent therapy is necessary to reduce resistance and improve the clinical outcome.


Colloid Carcinoma of the Uterine Cervix and Its Immunohistochemical Analysis: A Case Report.
Colloid carcinoma, which is a very rare tumor of the uterine cervix, is composed of an excessive amount of mucus and a relative paucity of tumoral glandular cells within them. Herein, we report a rare case of colloid carcinoma of the cervix with adenocarcinoma in situ (AIS), intestinal and usual types, and endocervical adenocarcinoma (usual type) components. We also discuss the morphological and immunohistochemical characteristics of this tumor. A 51-year-old woman was referred to our outpatient clinic with the symptom of genital bleeding lasting for 5 months. She had a cervix surrounded by an irregular tumor with a diameter of 5 cm. The colloid carcinoma cells were positive for MUC2, MUC5AC, and cytokeratin (CK) 7, focal positive for CDX2, and negative for MUC6 and CK20. Also, the intestinal type AIS showed a similar staining pattern. Colloid carcinoma cells producing mucin showed an intestinal phenotype and AIS. The intestinal type can be considered as a precursor lesion of colloid carcinoma.


The design of controllers for batch bioreactors.
The implementation of control algorithms to batch bioreactors is often complicated by variations in process dynamics that occur during the course of fermentation. Such a wide operating range often renders the performance of fixed gain proportional-integral-differential (PID) controllers unsatisfactory. In this work, detailed studies on the control of batch fermentations are per formed. Two simple controller designs are presented with the intent to compensate for changing process dynamics. One design incorporates the concepts of static feedforward-feedback control. While this technique produces tighter control than feedback alone, it is not as successful as a controller based on gain scheduling. The gain-scheduling controller, a subclass of adaptive controllers, uses the oxygen uptake rate as an auxiliary variable to fine-tune the PID controller parameters. The control of oxygen tension in the bioreactor is used as a vehicle to convey the proposed ideas, analyses, and results. Simulation experiments indicate significant improvement in controller performance can be achieved by both of the proposed approaches even in the presence of measurement noise.


Changing characteristics of livestock-associated meticillin-resistant Staphylococcus aureus isolated from humans - emergence of a subclade transmitted without livestock exposure, the Netherlands, 2003 to 2014.
Since 2007, livestock-associated meticillin-resistant Staphylococcus aureus (LA-MRSA) has become the predominant MRSA clade isolated from humans in the Netherlands. To assess possible temporal changes, we molecularly characterised over 9,000 LA-MRSA isolates submitted from 2003 to 2014 to the Dutch MRSA surveillance. After an initial rapid increase with a peak in 2009 (n = 1,368), the total number of submitted LA-MRSA isolates has been slowly decreasing to 968 in 2014 and over 80% of LA-MRSA belonged to one of three predominant MLVA/spa-types. Next generation sequencing (n=118) showed that MT569/t034 isolates were genetically more diverse than MT398/t011 and MT572/t108. Concurrent with the decrease in LA-MRSA, fewer people reported having contact with livestock and this was most prominent for people carrying MT569/t034 LA-MRSA. The proportion of LA-MRSA isolated from infection-related materials increased from 6% in 2009, to 13% in 2014 and most of these isolates originated from patients older than 50 years of age. Remarkably, 83% of these patients reported not having contact with livestock. The results reveal an ongoing change in the genotypic and epidemiological characteristics of Dutch LA-MRSA isolated from humans with the emergence of a LA-MRSA subclade independent of livestock exposure, suggesting LA-MRSA starts to resemble non-LA-MRSA in terms of transmissibility and pathogenicity.


[Study on the apoptosis of HL-60 human promyeloid leukaemia cells induced by SFPS].
The inhibition effects of Sargassum fusiforme Polysaccharides (SFPS) on proliferation of human HL-60 promyeloid leukemia cells were measured by MTT assay. SFPS-induced apoptosis in HL-60 cells was observed by electron microscopy, flow cytometry and DNA electrophoresis. The results revealed that SFPS exhibited antiproliferative activity which depended on dosage and time. After incubation for 24, 36, 48 and 72 h, SFPS had growth inhibitory effect on HL-60 cells with IC50 of 390, 362, 402 and 421 mg/L. A ladder-like pattern of DNA fragmentation was demonstrated on agarose gel electrophoresis after HL-60 cells induced by SFPS at a dose of 300 mg/L and 500 mg/L. Morphological examination by electron microscopy showed cells with few microvilli on the surface, chromatin condensation and margination, cell shrinkage and the presence of "apoptosis bodies". The DNA content analysis of FCM showed pre-G1 cells and increased G2 period ratio, in addition, it demonstrated that apoptosis induced by SFPS was dose-and time-dependent. In conclusion, the antitumor effect of SFPS is relative to cell apoptosis and the blocking of G2/M period cells.


Diascopy: a clinical technique for the diagnosis of vascular lesions.
Dentists can quickly determine the vascular nature of many intraoral lesions by applying a clinical test for blanchability. Such a clinical test is called a diascopy. Described here are the circumstances in which diascopy can be used in a dental practice and what useful information is provided by performing diascopy on oral mucosal lesions.


PROPELLER MRI visualizes detailed pathology of hippocampal sclerosis.
Hippocampal sclerosis (HS) is the most common cause of refractory temporal lobe epilepsy. Histopathologically, HS is characterized by neuron loss and gliosis. HS can be identified on MRI by signal increase on T(2)-weighted images and volume loss on T(1)-weighted volume images. The Periodically Rotated Overlapping Parallel Lines with Enhanced Reconstruction ("PROPELLER") sequence has excellent contrast between grey and white matter and compensates for subjects moving during the scan. The aim of the current report was to explore PROPELLER image quality of the hippocampus compared to routine sequences. Routine sequences (T(1) volume, T(2)-weighted, PD and FLAIR images) and PROPELLER images were acquired in four presurgical patients with HS using a GE 3T Excite HD scanner (General Electric). Resected tissue was stained with LFB, and for GFAP, NeuN and dynorphin immunohistochemistry. Hippocampal sections were compared with PROPELLER images. PROPELLER images were T(2)-weighted and had superior tissue contrast compared to routine sequences. PROPELLER images showed the internal hippocampal structures and tissue changes associated with HS. This corresponded to changes seen on histopathological sections confirming that the sequence could distinguish between different strata and subfields of the hippocampus. The PROPELLER sequence shows promise for detailed in vivo imaging of the hippocampus in patients who did not move overtly, negating the inevitable subtle movements during scans. More detailed in vivo studies of the hippocampal formation, investigating subtle abnormalities such as end folium sclerosis, and the neocortex are now possible and may increase the diagnostic yield of MRI.


Association between chloroplast DNA and mitochondrial DNA haplotypes in Prunus spinosa L. (Rosaceae) populations across Europe.
Chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) were studied in 24 populations of Prunus spinosa sampled across Europe. The cpDNA and mtDNA fragments were amplified using universal primers and subsequently digested with restriction enzymes to obtain the polymorphisms. Combinations of all the polymorphisms resulted in 33 cpDNA haplotypes and two mtDNA haplotypes. Strict association between the cpDNA haplotypes and the mtDNA haplotypes was detected in most cases, indicating conjoint inheritance of the two genomes. The most frequent and abundant cpDNA haplotype (C20; frequency, 51 %) is always associated with the more frequent and abundant mtDNA haplotype (M1; frequency, 84 %). All but two of the cpDNA haplotypes associated with the less frequent mtDNA haplotype (M2) are private haplotypes. These private haplotypes are phylogenetically related but geographically unrelated. They form a separate cluster on the minimum-length spanning tree.


Warburg effect-related risk scoring model to assess clinical significance and immunity characteristics of glioblastoma.
Glioblastoma (GBM), the most common primary malignant brain tumor, has a poor prognosis, with a median survival of only 14.6 months. The Warburg effect is an abnormal energy metabolism, which is the main cause of the acidic tumor microenvironment. This study explored the role of the Warburg effect in the prognosis and immune microenvironment of GBM. A prognostic risk score model of Warburg effect-related genes (Warburg effect signature) was constructed using GBM cohort data from The Cancer Genome Atlas. Cox analysis was performed to identify independent prognostic factors. Next, the nomogram was built to predict the prognosis for GBM patients. Finally, the drug sensitivity analysis was utilized to find the drugs that specifically target Warburg effect-related genes. Age, radiotherapy, chemotherapy, and WRGs score were confirmed as independent prognostic factors for GBM by Cox analyses. The C-index (0.633 for the training set and 0.696 for the validation set) and area under curve (>0.7) indicated that the nomogram exhibited excellent performance. The calibration curve also indicates excellent consistency of the nomogram between predictions and actual observations. In addition, immune microenvironment analysis revealed that patients with high WRGs scores had high immunosuppressive scores, a high abundance of immunosuppressive cells, and a low response to immunotherapy. The Cell Counting Kit-8 assays showed that the drugs targeting Warburg effect-related genes could inhibit the GBM cells growth in vitro. Our research showed that the Warburg effect is connected with the prognosis and immune microenvironment of GBM. Therefore, targeting Warburg effect-related genes may provide novel therapeutic options.


Studying risk factors for skin cancer development.
In most countries with a population of mainly European origin, non-melanoma skin carcinoma incidence rate has increased over the last decade. Understanding of what is the best way to identify people at high risk for skin cancer development will help to optimize the strategy for prevention and treatment tactics. Aim of the research involves studying risk factors for skin cancer development in patients with cutaneous squamous cell carcinoma and squamous cell carcinoma in situ (cSCC and SCCis correspondingly) and patients with actinic keratosis (AK). we conducted a survey of patients with cSCC, SCCis and AK who had been under case follow-up at the State Scientific Institution "Scientific and Practical Centre of Preventive and Clinical Medicine" of the State Administration (hereinafter SIS) during 2014-2016.Clinical diagnosis in 100% of cases was confirmed by pathomorphological study. Statistical processing of the obtained data was performed using STATISTICA 7.0 programme. The questionnaire involved 129 patients, including 21(16.3%) patient with cSCC, 40 (31%) patients with SCCis, 57 (44.2%) patients with AK and 11 (8.5%) patients who had mentioned above combined pathology i.e. cSCC, SCCis and АК at the same time. Only two studied risk factors for skin cancer development were found to have influence on the skin pathology: "Not using sun-protection preparation for the skin" and "Burdened family history of thefirst level of relationship". In forming the risk groups for skin cancer development it is advisable to consider burdened family history and using sun-protection preparation for the skin.


Integrated lithium niobate optical phased array for two-dimensional beam steering.
Optical phased arrays (OPAs) with high speed, low power consumption, and low insertion loss are appealing for various applications, including light detection and ranging, free-space communication, image projection, and imaging. These OPAs can be achieved by fully harnessing the advantages of integrated lithium niobate (LN) photonics, which include high electro-optical modulation speed, low driving voltage, and low optical loss. Here we present an integrated LN OPA that operates in the near-infrared regime. Our experimental results demonstrate 24 × 8° two-dimensional beam steering, a far-field beam spot with a full width at half maximum of 2 × 0.6°, and a sidelobe suppression level of 10 dB. Furthermore, the phase modulator of our OPA exhibits a half-wave voltage of 6 V. The low power consumption exhibited by our OPA makes it highly attractive for a wide range of applications. Beyond conventional applications, our OPA's high speed opens up the possibility of novel applications such as high-density point cloud generation and tomographic holography.


Modulation of external pancreatic secretion by endogenous norepinephrine: study with a norepinephrine uptake blocker in the rat.
The effect of endogenous catecholamines on pancreatic secretion was analyzed with nisoxetine, a specific norepinephrine uptake blocker, and specific adrenoceptor antagonists in anesthetized acute fistula rats. Nisoxetine was administered alone or with alpha-1 (prazosin), alpha-2 (idazoxan or yohimbine), or beta (propranolol) adrenoceptor antagonists. Pancreatic secretion was measured in basal conditions or after stimulation by 2-deoxy-D-glucose (2DG), electrical vagal stimulation, or acetylcholine. (a) Basal. Nisoxetine alone had no effect. Associated with idazoxan or yohimbine, nisoxetine produced a dose-related stimulation (p < 0.01) of water and electrolyte without changing protein output. Addition of propranolol abolished this stimulation. (b) 2DG. Nisoxetine inhibited 2DG-induced secretion (p < 0.01). Idazoxan or yohimbine suppressed the nisoxetine inhibition of water and electrolyte output (p < 0.01) but had no effect on protein output, which was restored only by adding a mixture of idazoxan, prazosin, and propranolol. (c) Electrical stimulation. Nisoxetine did not modify water and electrolyte but inhibited protein response by 75%. Adding idazoxan to nisoxetine significantly increased (p < 0.01) water and bicarbonate response and partly restored protein response. Water and bicarbonate response was restored by propranolol, whereas protein response was only restored by adding a mixture of idazoxan, prazosin, and propranolol. (d) Nisoxetine did not modify pancreatic response to acetylcholine. In conclusion, endogenous norepinephrine affects the response to vagally mediated effects through several subtypes of adrenoceptors, without changing basal or acetylcholine stimulated secretion.


Extracellular vesicles secreted by highly metastatic clonal variants of osteosarcoma preferentially localize to the lungs and induce metastatic behaviour in poorly metastatic clones.
Osteosarcoma (OS) is the most common pediatric bone tumor and is associated with the emergence of pulmonary metastasis. Unfortunately, the mechanistic basis for metastasis remains unclear. Tumor-derived extracellular vesicles (EVs) have been shown to play critical roles in cell-to-cell communication and metastatic progression in other cancers, but their role in OS has not been explored. We show that EVs secreted by cells derived from a highly metastatic clonal variant of the KHOS cell line can be internalized by a poorly metastatic clonal variant of the same cell line and induce a migratory and invasive phenotype. This horizontal phenotypic transfer is unidirectional and provides evidence that metastatic potential may arise via interclonal co-operation. Proteomic analysis of the EVs secreted by highly metastatic OS clonal variants results in the identification of a number of proteins and G-protein coupled receptor signaling events as potential drivers of OS metastasis and novel therapeutic targets. Finally, multiphoton microscopy with fluorescence lifetime imaging in vivo, demonstrated a preferential seeding of lung tissue by EVs derived from highly metastatic OS clonal variants. Thus, we show that EVs derived from highly metastatic clonal variants of OS may drive metastatic behaviour via interclonal co-operation and preferential colonization of the lungs.


Proteomic analysis of tears following acupuncture treatment for menopausal dry eye disease by two-dimensional nano-liquid chromatography coupled with tandem mass spectrometry.
The purpose of this study was to investigate whether acupuncture is effective at treating dry eye disease among postmenopausal women and to identify the possible mechanisms. Twenty-eight postmenopausal women with dry eye disease were randomly divided into two groups: an acupuncture plus artificial tears (AC + AT) group and an artificial tears (AT) only group. After baseline examination of clinical parameters and tear sample collection, each patient received the designated modality of topical therapy for 2 months. Post-treatment documentation of clinical parameters was recorded, and tear samples were collected. Tear samples from the AC + AT group were subjected to two-dimensional nano-liquid chromatography coupled with tandem mass spectrometry (2D nano-LC-MS/MS). Western blot analysis was also performed on tear samples from both groups. After treatment, the Ocular Surface Disease Index scores, symptom assessment scores, scores of sign assessment, and tear break-up time were significantly improved in both groups (P=0.000). Symptom assessment scores were significantly improved in the AC + AT group (P=0.000) compared with the AT group. 2D nano-LC-MS/MS identified 2,411 proteins, among which 142 were downregulated and 169 were upregulated. After combined AC + AT treatment, the abundance of secreted proteins was increased, whereas that of cytoplasmic proteins decreased (Pearson's χ2 test, P=0.000, P=0.000, respectively). Proteins involved in immunity and regulation were also more abundant (Pearson's χ2 test, P=0.040, P=0.016, respectively), while components and proliferation-related proteins were downregulated (Pearson's χ2 test, P=0.003, P=0.011, respectively). AC + AT treatment increased protein synthesis and secretion, and improved clinical symptoms. These results indicate that acupuncture may be a complimentary therapy for treating postmenopausal dry eye disease.


Active Instrument Engagement Combined with a Real-Time Database Search for Improved Performance of Sample Multiplexing Workflows.
Quantitative proteomics employing isobaric reagents has been established as a powerful tool for biological discovery. Current workflows often utilize a dedicated quantitative spectrum to improve quantitative accuracy and precision. A consequence of this approach is a dramatic reduction in the spectral acquisition rate, which necessitates the use of additional instrument time to achieve comprehensive proteomic depth. This work assesses the performance and benefits of online and real-time spectral identification in quantitative multiplexed workflows. A Real-Time Search (RTS) algorithm was implemented to identify fragment spectra within milliseconds as they are acquired using a probabilistic score and to trigger quantitative spectra only upon confident peptide identification. The RTS-MS3 was benchmarked against standard workflows using a complex two-proteome model of interference and a targeted 10-plex comparison of kinase abundance profiles. Applying the RTS-MS3 method provided the comprehensive characterization of a 10-plex proteome in 50% less acquisition time. These data indicate that the RTS-MS3 approach provides dramatic performance improvements for quantitative multiplexed experiments.


Decreased uptake of 3H-serotonin and endogenous content of serotonin in blood platelets in hypertensive patients.
The uptake and content of serotonin in blood platelets were studied in patients with essential hypertension and in five families in which at least one member was hypertensive. Blood was obtained from male and female normotensive volunteers and hypertensive patients who were free of medication. Lineweaver-Burk plots of 3H-serotonin uptake from both control subjects and hypertensive patients were linear, which suggested simple Michaelis-Menten uptake kinetics. The maximal uptake velocity (Vmax) in hypertensive patients was significantly lower than in control subjects (control = 41.7 +/- 3.3 pmol/min/10(8) platelets, n = 17; hypertensive = 26.6 +/- 3.0 pmol/min/10(8) platelets, n = 16; p less than 0.005). The affinity constant (Km) was slightly but significantly lower in hypertensive patients (control = 0.70 +/- 0.08 microM; hypertensive = 0.46 +/- 0.08 microM; p less than 0.05). The serotonin content in blood platelets determined by high pressure liquid chromatography with electrochemical detection was significantly lower in hypertensive patients (control = 165.0 +/- 12.9 nmol/10(11) platelets, n = 29; hypertensive = 105.9 +/- 10.4 nmol/10(11) platelets, n = 27; p less than 0.001). In the five families investigated, the lowered serotonin content was observed in some normotensive members. The reduced number of carriers of serotonin uptake and the slight decrease in the affinity constant observed in platelets of patients with essential hypertension suggest that serotonin metabolism is altered in essential hypertension and that blood platelets may be a useful model in studying the serotonergic modifications at the molecular level.


Molecular evidence for conspecificity of two desert Liolaemus lizards (Iguania: Liolaemidae).
Liolaemus audituvelatus (Núñez Yáñez 1983) and L. manueli (Núñez, Navarro, Garín, Pincheira-Donoso Meriggio 2003) are endemic species of the Atacama Desert of northern Chile that belong to the montanus group. Both species are considered cryptic from each other and can only be distinguished by their distribution ranges and karyotypes. Originally, there was a wide separation zone between their known distribution ranges, but later collections reduced the gap from 430 km to only 150 km. In this study, we review the geographic information about both species and report new localities within the distribution gap, where species identification becomes difficult. We performed a molecular phylogenetic analysis and applied several species delimitation methods to reassess the taxonomic status of both nominal species and new intermediate populations. Our analyses support the placement of L. manueli in the synonymy of L. audituvelatus. We discuss the biogeographic and conservation implications of this new synonymy.


Molecular, biochemical characterization and assessment of immunogenic potential of cofactor-independent phosphoglycerate mutase against Leishmania donovani: a step towards exploring novel vaccine candidate.
Despite immense efforts, vaccine against visceral leishmaniasis has yet not been developed. Earlier our proteomic study revealed a novel protein, cofactor-independent phoshoglycerate mutase (LdiPGAM), an important enzyme in glucose metabolism, in T helper cells type 1 (Th1) stimulatory region of soluble Leishmania donovani antigen. In this study, LdiPGAM was biochemically and molecularly characterized and evaluated for its immunogenicity and prophylactic efficacy against L. donovani. Immunogenicity of recombinant LdiPGAM (rLdiPGAM) was initially assessed in naïve hamsters immunized with it by analysing mRNA expression of inducible nitric oxide (NO) synthase (iNOS) and other Th1/T helper cells type 2 cytokines, which revealed an upregulation of Th1 cytokines along with iNOS. Immunogenicity of rLdiPGAM was further evaluated in lymphocytes of treated Leishmania-infected hamsters and peripheral blood mononuclear cells of Leishmania patients in clinical remission by various parameters, viz. lymphoproliferation assay and NO production (hamsters and patients) and levels of various cytokines (patients). rLdiPGAM induced remarkable Lymphoproliferative response and NO production in treated Leishmania-infected hamsters as well as in patients and increase in interferon gamma (IFN-γ), interleukin-12 (IL-12p40) responses in Leishmania patients in clinical remission. Vaccination with rLdiPGAM exerted considerable prophylactic efficacy (73%) supported by increase in mRNA expression of iNOS, IFN-γ and IL-12p40 with decrease in transforming growth factor beta and interleukin-10. Above results indicate the importance of rLdiPGAM protein as a potential vaccine candidate against visceral leishmaniasis.


LC3-Associated Endocytosis Facilitates β-Amyloid Clearance and Mitigates Neurodegeneration in Murine Alzheimer's Disease.
The expression of some proteins in the autophagy pathway declines with age, which may impact neurodegeneration in diseases, including Alzheimer's Disease. We have identified a novel non-canonical function of several autophagy proteins in the conjugation of LC3 to Rab5+, clathrin+ endosomes containing β-amyloid in a process of LC3-associated endocytosis (LANDO). We found that LANDO in microglia is a critical regulator of immune-mediated aggregate removal and microglial activation in a murine model of AD. Mice lacking LANDO but not canonical autophagy in the myeloid compartment or specifically in microglia have a robust increase in pro-inflammatory cytokine production in the hippocampus and increased levels of neurotoxic β-amyloid. This inflammation and β-amyloid deposition were associated with reactive microgliosis and tau hyperphosphorylation. LANDO-deficient AD mice displayed accelerated neurodegeneration, impaired neuronal signaling, and memory deficits. Our data support a protective role for LANDO in microglia in neurodegenerative pathologies resulting from β-amyloid deposition.


Towards a true neural stance on consciousness.
Consciousness is traditionally defined in mental or psychological terms. In trying to find its neural basis, introspective or behavioral observations are considered the gold standard, to which neural measures should be fitted. I argue that this poses serious problems for understanding the mind-brain relationship. To solve these problems, neural and behavioral measures should be put on an equal footing. I illustrate this by an example from visual neuroscience, in which both neural and behavioral arguments converge towards a coherent scientific definition of visual consciousness. However, to accept this definition, we need to let go of our intuitive or psychological notions of conscious experience and let the neuroscience arguments have their way. Only by moving our notion of mind towards that of brain can progress be made.


[Steroid pulse therapy].
Intravenous methylprednisolone pulse (IVMP) is usually given because of its powerful and rapid immunosuppressive effect. In a randomized control trial for all patients with Kawasaki disease(KD), IVMP plus initial immune globulin(IVIG), compared with IVIG plus placebo, did not decrease the incidence of coronary artery lesions(CAL). It has been reported that suspected IVIG-resistant patients who received initial IVIG plus IVMP, compared with IVIG alone, had earlier defervescence and a significantly lower rate of CAL. For patients resistant to initial or additional IVIG, some studies showed that IVMP was effective for fast defervescence and prevention of CAL. As it stands, IVMP is indicated for initial therapy combined with IVIG for suspected IVIG-resistant patients or rescue therapy for IVIG-resistant patients.


Fume levels during bone cement mixing.
This article compares some of the various products available for mixing and preparing bone cement. It should be noted that the tests were not independent as the author is employed by one of the product manufacturers. It should also be noted that the tests were carried out under laboratory conditions and therefore do not include practices carried out by scrubbed staff that could affect the results.


Exposure to Umbelliferone Reduces Ralstonia solanacearum Biofilm Formation, Transcription of Type III Secretion System Regulators and Effectors and Virulence on Tobacco.
Ralstonia solanacearum is one of the most devastating phytopathogens and causes bacterial wilt, which leads to severe economic loss due to its worldwide distribution and broad host range. Certain plant-derived compounds (PDCs) can impair bacterial virulence by suppressing pathogenic factors of R. solanacearum. However, the inhibitory mechanisms of PDCs in bacterial virulence remain largely unknown. In this study, we screened a library of coumarins and derivatives, natural PDCs with fused benzene and α-pyrone rings, for their effects on expression of the type III secretion system (T3SS) of R. solanacearum. Here, we show that umbelliferone (UM), a 7-hydroxycoumarin, suppressed T3SS regulator gene expression through HrpG-HrpB and PrhG-HrpB pathways. UM decreased gene expression of six type III effectors (RipX, RipD, RipP1, RipR, RipTAL, and RipW) of 10 representative effector genes but did not alter T2SS expression. In addition, biofilm formation of R. solanacearum was significantly reduced by UM, though swimming activity was not affected. We then observed that UM suppressed the wilting disease process by reducing colonization and proliferation in tobacco roots and stems. In summary, the findings reveal that UM may serve as a plant-derived inhibitor to manipulate R. solanacearum T3SS and biofilm formation, providing proof of concept that these key virulence factors are potential targets for the integrated control of bacterial wilt.


Thrombotic Microangiopathy after a 15-year Treatment with Interferon Beta-1b in a Patient with Multiple Sclerosis: A Case Report and Review of Literature.
A 54-year-old woman with multiple sclerosis treated with interferon-β (IFN-β)-1b for 15 years presented with sustained hypertension (240/124 mmHg) and retinal bleeding. She had proteinuria, anemia, thrombocytopenia, elevated serum creatinine levels, and haptoglobin depletion. Intravenous nicardipine stabilized her blood pressure, but her renal function and platelet count deteriorated. The initial disintegrin-like metalloprotease with thrombospondin type 1 motif 13 (ADAMTS13) activity was 28% of normal without its inhibitor. The subsequent peripheral appearance of schistocytes suggested thrombotic microangiopathy (TMA). After IFN-β-1b cessation, the platelet count increased, and the blood pressure stabilized. The ADAMTS13 activity normalized, although the creatinine level did not. TMA may develop after the long-term use of IFN-β without adverse events.


Simulation-based low-dose, high-frequency plus mobile mentoring versus traditional group-based trainings among health workers on day of birth care in Nigeria; a cluster randomized controlled trial.
The aim of this study was to compare health workers knowledge and skills competencies between those trained using the onsite simulation-based, low-dose, high frequency training plus mobile mentoring (LDHF/m-mentoring) and the ones trained through traditional offsite, group-based training (TRAD) approach in Kogi and Ebonyi states, Nigeria, over a 12-month period. A prospective cluster randomized controlled trial was conducted by enrolling 299 health workers who provided healthcare to mothers and their babies on the day of birth in 60 health facilities in Kogi and Ebonyi states. These were randomized to either LDHF/m-mentoring (intervention, n = 30 facilities) or traditional group-based training (control, n = 30 facilities) control arm. They received Basic Emergency Obstetrics and Newborn Care (BEmONC) training with simulated practice using anatomic models and role-plays. The control arm was trained offsite while the intervention arm was trained onsite where they worked. Mentorship was done through telephone calls and reminder text messages. The multiple choice questions (MCQs) and objective structured clinical examinations (OSCEs) mean scores were compared; p-value < 0.05 was considered statistically significant. Qualitative data were also collected and content analysis was conducted. The mean knowledge scores between the two arms at months 3 and 12 post-training were equally high; no statistically significant differences. Both arms showed improvements in composite scores for assessed BEmONC clinical skills from around 30% at baseline to 75% and above at end line (p < 0.05). Overall, the observed improvement and retention of skills was higher in intervention arm compared to the control arm at 12 months post-training, (p < 0.05). Some LDHF/m-mentoring approach trainees reported that mentors' support improved their acquisition and maintenance of knowledge and skills, which may have led to reductions in maternal and newborn deaths in their facilities. The LDHF/m-mentoring intervention is more effective than TRAD approach in improving health workers' skills acquisition and retention. Health care managers should have the option to select the LDHF/m-mentoring learning approach, depending on their country's priorities or context, as it ensures health workers remain in their place of work during training events thus less disruption to service delivery. The trial was retrospectively registered on August 24, 2017 at ClinicalTrials.Gov: NCT03269240.


Nurses' knowledge of infection control.
The article is based on a research study examining infection control in nurse education and practice. A survey of a large population was carried out to establish the perceived importance of microbiological knowledge to nurses and to ascertain whether this knowledge was present. The results suggest that although microbiological knowledge is considered necessary for safe infection control practice, nurses' actual knowledge falls far short of the level required for 'informed' practice. What this implies in relation to patient care, and recommendations regarding education and practice, are discussed.


Lennox-Gastaut syndrome. Clinical description and diagnosis.
The Lennox-Gastaut syndrome (LGS) is widely known as one of the most severe and prognostically unfavourable epileptic conditions. Its entity as a syndrome is based on the occurrence of certain characteristic types of seizure (particularly typical drop attacks and axial tonic seizures) and on EEG patterns such as the slow spike-wave complex and runs of rapid spikes (10-25/sec).


Detailed simulation of the response function of an Si(Li) detector.
The response function of an Si(Li) detector was calculated by using a Monte Carlo method for photon energies below 10 keV. In the simulation, electron elastic-scattering cross sections calculated using the partial-wave expansion method were used instead of the Born approximation to simulate the electron scattering more accurately. The carrier collection probability near the gold-silicon interface was derived by solving the carrier continuity equation with drift, diffusion and surface recombination by a finite recombination velocity. For two detectors, good agreement was found for low-energy tailing between measured and simulated response functions. The integrated detection efficiency is almost identical with the transmittance in the front gold electrode showing a maximum difference of 2% at the Au M(4)-absorption edge.


Impact of prolactin on epididymal lipid profile in castrated rats.
Prolactin treatment to castrated rats led to accumulation of triacylglycerol and esterified cholesterol. There was no appreciable drift in epididymal cholesterol: phospholipid ratio between the prolactin treated and control animals. However, further analysis of phospholipids showed a build up of phosphatidyl inositol, phosphatidyl choline and phosphatidyl ethanolamine but a drop in the levels of phosphatidyl serine and sphingomyelin in prolactin treated castrated rats as compared to those castrated animals injected with vehicle alone. Changes in phospholipids reported above were prominently seen in the group of castrated rats that received 100 micrograms oPRL/100 g body weight but not in those animals which received either lower or higher doses of the hormone. Interestingly, bromocryptine treatment in castrated rats produced a general depletion in the levels of all lipid classes studied in the epididymis. It is suggested that this may be due to impaired synthesis and/or increased breakdown of lipids in this organ.


Implementing objective structured clinical skills evaluation (OSCE) in nurse registration programmes in a centre in Ireland: a utilisation focused evaluation.
This paper presents findings from a research study of two objective structured clinical skills evaluation (OSCE) processes that were used in the new BSc. in General and Psychiatric Nursing Registration programmes offered in an Institute of Technology in the south west of Ireland. OSCEs, which have gained widespread acceptance throughout the western world as valid academic measures of nursing competence has yet to be established in this country. This research aimed to evaluate the process and outcomes of OSCE from the perspective of the major stakeholder groups: first and second year nursing students, lecturers, clinical placement coordinators and assessors. A utilisation focussed evaluation approach was adopted which values the opinions and experiences of stakeholders. It is intended to use the findings to develop the validity and quality of the OSCE assessment. The OSCE process was found to have a positive impact on all stakeholders. OSCEs were perceived to be a meaningful and fair form of assessment. Students identified that they felt more prepared for and more confident about forthcoming placements. The OSCE process was, however, perceived to be a stressful experience and requires considerable preparation effort by students and academic staff. Mature students claimed that more practice effort was required but also felt more prepared for placements and achieved higher OSCE scores.


Development of Omni InDel and supporting database for maize.
Insertions-deletions (InDels) are the second most abundant molecular marker in the genome and have been widely used in molecular biology research along with simple sequence repeats (SSR) and single-nucleotide polymorphisms (SNP). However, InDel variant mining and marker development usually focuses on a single type of dimorphic InDel, which does not reflect the overall InDel diversity across the genome. Here, we developed Omni InDels for maize, soybean, and rice based on sequencing data and genome assembly that included InDel variants with base lengths from 1 bp to several Mb, and we conducted a detailed classification of Omni InDels. Moreover, we screened a set of InDels that are easily detected and typed (Perfect InDels) from the Omni InDels, verified the site authenticity using 3,587 germplasm resources from 11 groups, and analyzed the germplasm resources. Furthermore, we developed a Multi-InDel set based on the Omni InDels; each Multi-InDel contains multiple InDels, which greatly increases site polymorphism, they can be detected in multiple platforms such as fluorescent capillary electrophoresis and sequencing. Finally, we developed an online database website to make Omni InDels easy to use and share and developed a visual browsing function called "Variant viewer" for all Omni InDel sites to better display the variant distribution.


Left ventricular hypertrophy in athletes and hypertensive patients.
Systemic hypertension and physical exercise are both associated with cardiac adaptations. The impact is most prominent on the left side of the heart, which hypertrophies leading to left ventricular hypertrophy. This article reviews structural and functional cardiac changes seen in hypertensive and athlete's hearts.


Bone marrow core biopsy specimens in AL (primary) amyloidosis. A morphologic and immunohistochemical study of 100 cases.
We retrospectively reviewed 100 bone marrow core biopsy specimens from patients with AL (primary) amyloidosis. The morphologic and immunohistochemical features were assessed by standard histochemical stains (H&E, periodic acid-Schiff, Congo red) and immunohistochemical stains for light chain immunoglobulins. Bone marrow core biopsy revealed a plasma cell dyscrasia in 83% (lambda, 65; kappa, 18) of cases. Amyloid deposits were observed in 60% of the bone marrow core biopsy specimens and, when present, were detected most often in blood vessel walls only (39/60). However, if present, interstitial amyloid deposition was significantly more associated with patients with a monoclonal kappa light chain gammopathy (P = .04). Through the careful analysis of standard histochemical and immunohistochemical stains, bone marrow core biopsy provides essential diagnostic information in cases of AL amyloidosis.


A Longitudinal Assessment of the Effect of Resident-Centered Care on Quality in Veterans Health Administration Community Living Centers.
To examine whether changes in resident-centered care (RCC) over time were associated with changes in quality. Data sources were the Minimum Dataset quality indicators (which consist of measures of both prevalence and incidence of adverse events) and the Artifacts of Culture Change Tool (which measures RCC; FYs 2009-2012) from 130 Veterans Health Administration community living centers. A retrospective longitudinal study. Data were from VA secondary data sources. The overall relationship between RCC and quality was not statistically significant (p = .22), although there was a weakly significant negative relationship (i.e., increased RCC was associated with poorer quality) in the seven quarters after implementation of an automated version of the Artifacts Tool (p = .08). In facility-specific analyses, there were 15 facilities with a weakly significant (p < .10) positive relationship between RCC and quality and 21 with a weakly significant negative relationship. Adjusted cost per patient day was over 50 percent higher in the 21 facilities with a negative relationship than in the 15 facilities with a positive relationship (p < .05). The Artifacts score is a formal performance metric in the VA, and thus, facilities were explicitly incentivized to increase RCC. Using qualitative methods to identify characteristics that distinguished those facilities able to increase both RCC and quality from those that suffered declines in quality as RCC was improved is an important follow-up to this study.


JNK pathway plays a key role in the immune system of the pea aphid and is regulated by microRNA-184.
Different from holometabolous insects, the hemipteran species such as pea aphid Acyrthosiphon pisum exhibit reduced immune responses with the absence of the genes coding for antimicrobial peptide (AMP), immune deficiency (IMD), peptidoglycan recognition proteins (PGRPs), and other immune-related molecules. Prior studies have proved that phenoloxidase (PO)-mediated melanization, hemocyte-mediated phagocytosis, and reactive oxygen species (ROS) participate in pea aphid defense against bacterial infection. Also, the conserved signaling, Jun N-terminal kinase (JNK) pathway, has been suggested to be involved in pea aphid immune defense. However, the precise role of the JNK signaling, its interplay with other immune responses and its regulation in pea aphid are largely unknown. In this study, using in vitro biochemical assays and in vivo bioassays, we demonstrated that the JNK pathway regulated hemolymph PO activity, hydrogen peroxide concentration and hemocyte phagocytosis in bacteria infected pea aphids, suggesting that the JNK pathway plays a central role in regulating immune responses in pea aphid. We further revealed the JNK pathway is regulated by microRNA-184 in response to bacterial infection. It is possible that in common the JNK pathway plays a key role in immune system of hemipteran insects and microRNA-184 regulates the JNK pathway in animals.


Effect of a single neonatal treatment with steroid hormone or steroid-like molecules on myocardial ouabain binding in the adult rat.
A single neonatal treatment of rats with vitamin D3, gibberellin, allylestrenol or diethylstilbestrol (DES) influenced the ouabain binding capacity of myocardial Na, K-dependent ATP-ase. Of the active molecules tested, vitamin D3, DES and gibberellin had appreciable impact on myocardial ouabain receptors, enhancing and depressing their activity, respectively. The thymic dexamethasone and uterine estrogen receptors did not alter their binding capacity in response to neonatal exposure to vitamin D3 or gibberellin.


[Changes in pathology since the times of Purkinje].
The discipline of pathology was founded in the Purkinje era on the principles of building scientific knowledge of diseases through co-operation among clinical specialists. During the 20th century the focus of pathologists work has progressed from autopsies to biopsy diagnostics. The discipline of histopathological and cytopathological biopsy took form and continuously incorporates new methodologies and new standards. Its future course compels us to reflect on the changing requirements of clinical medicine. Current undergraduate education programs are directed toward forming a basis of expertise for both clinicians and non-medical healthcare professionals. At the postgraduate level and in research we are seeing an ever-closer interdisciplinary link, especially within the fields of genetics. This approach brings pathological processes to a new level, but also demands new levels of knowledge. Through comprehensive diagnostic experience, both prognostic and predictive, a pathologist is an indispensable member of the diagnostic and therapeutic team.


Gastrointestinal cell lines form polarized epithelia with an adherent mucus layer when cultured in semi-wet interfaces with mechanical stimulation.
Mucin glycoproteins are secreted in large quantities by mucosal epithelia and cell surface mucins are a prominent feature of the glycocalyx of all mucosal epithelia. Currently, studies investigating the gastrointestinal mucosal barrier use either animal experiments or non-in vivo like cell cultures. Many pathogens cause different pathology in mice compared to humans and the in vitro cell cultures used are suboptimal because they are very different from an in vivo mucosal surface, are often not polarized, lack important components of the glycocalyx, and often lack the mucus layer. Although gastrointestinal cell lines exist that produce mucins or polarize, human cell line models that reproducibly create the combination of a polarized epithelial cell layer, functional tight junctions and an adherent mucus layer have been missing until now. We trialed a range of treatments to induce polarization, 3D-organization, tight junctions, mucin production, mucus secretion, and formation of an adherent mucus layer that can be carried out using standard equipment. These treatments were tested on cell lines of intestinal (Caco-2, LS513, HT29, T84, LS174T, HT29 MTX-P8 and HT29 MTX-E12) and gastric (MKN7, MKN45, AGS, NCI-N87 and its hTERT Clone5 and Clone6) origins using Ussing chamber methodology and (immuno)histology. Semi-wet interface culture in combination with mechanical stimulation and DAPT caused HT29 MTX-P8, HT29 MTX-E12 and LS513 cells to polarize, form functional tight junctions, a three-dimensional architecture resembling colonic crypts, and produce an adherent mucus layer. Caco-2 and T84 cells also polarized, formed functional tight junctions and produced a thin adherent mucus layer after this treatment, but with less consistency. In conclusion, culture methods affect cell lines differently, and testing a matrix of methods vs. cell lines may be important to develop better in vitro models. The methods developed herein create in vitro mucosal surfaces suitable for studies of host-pathogen interactions at the mucosal surface.


Ingenol-3-Angelate Suppresses Growth of Melanoma Cells and Skin Tumor Development by Downregulation of NF-κB-Cox2 Signaling.
BACKGROUND A recent focus in skin cancer prevention intervenes though modulating molecular links between inflammation and cell growth signaling, such as NF-κB. This study elucidates the effect of a non-tumor promoting phorbol ester, ingenol-3-angelate (I3A), on the growth of human melanoma cells and on the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced skin inflammation and 7,12-Dimethylbenz(a)anthracene (DMBA)-induced skin carcinoma in mice. MATERIAL AND METHODS Cell viability was assessed by MTT assay, cell proliferation by clonogenic assay, apoptosis and cell cycle arrest was analyzed by flow cytometry, protein expression was studied by IHC and Western blotting, and gene expression by qPCR. RESULTS I3A suppressed the survival and proliferation of human melanoma cells with estimated IC50 values around 38 and 46 μM for A2058 and HT144 cell, respectively. I3A activated the protein levels of PKCδ and PKCε, which induced apoptosis by activating caspase-9 and caspace-3 followed by lowering of mitochondrial membrane potential and enhancing DNA fragmentation. I3A induced G1 phase cell cycle arrest as well as G2/M phase arrest in both cell lines. I3A inhibited the levels of NFκB p65 protein as well as phosphorylation of p65 and its nuclear translocation. I3A suppressed the gene expression of NF-κB, COX-2 and iNOS. I3A inhibited TPA-induced inflammation and epidermal hyperplasia in female ICR mice by downregulating NF-κB and iNOS. I3A suppressed the growth of skin tumor in DMBA-induced mice in dose-dependent manner. CONCLUSIONS The mechanism of I3A induces apoptosis in human melanoma cells and suppresses skin inflammation and carcinoma via downregulation of NF-κB-iNOS-COX-2 signaling.


Clinical and electrophysiological aspects of distal ulnar neuropathy.
To evaluate the use of fixed distance side to side comparison of abductor digiti minimi (ADM) and first dorsal interosseous (FDI) compound muscle action potential (CMAP) studies in the diagnosis of distal ulnar neuropathy. Thirty normal controls underwent ADM and FDI CMAP studies bilaterally at a fixed ADM recording to stimulating point distance of 6 cm. Side to side mean latency differences to both muscles were calculated. Twenty patients with suspected unilateral distal ulnar neuropathy from routine nerve conduction studies had positive results when compared with controls. The upper limit of normal for side to side mean latency difference at 3 SD above the mean was 0.394 and 0.474 ms for ADM and FDI, respectively. All 20 patients had side to side mean CMAP latency difference above 3 SDs in the ADM, FDI or both muscles. Fifty percent of cases had involvement of the superficial sensory branch. Fixed distance side to side CMAP latency comparison is a useful electrodiagnostic adjunct for distal ulnar neuropathy. Trauma was the most common aetiology in the 20 reported cases. Correlation was found between aetiological factors and sites of lesions as localized with this method.


Parasite performance and host alternation: is there a negative effect in host-specific and host-opportunistic parasites?
Environmental fluctuations are expected to require special adaptations only if they are associated with a decrease in fitness. We compared reproductive performance between fleas fed on alternating (preferred and non-preferred) hosts and fleas fed solely on either a preferred or a non-preferred host to determine whether (1) host alternation incurs an immediate negative effect, and, if yes, then (2) whether this effect is greater in a host specialist (Parapulex chephrenis) than in host generalists (Xenopsylla conformis and Synosternus cleopatrae). We also compared flea performance under alternating host regimes with different host order (initial feeding on either a preferred or a non-preferred host). An immediate negative effect of alternating hosts on reproductive performance was found in P. chephrenis only. These fleas produced 44·3% less eggs that were 3·6% smaller when they fed on alternating hosts as compared with a preferred host. In contrast, X. conformis and S. cleopatrae appeared to be able to adapt their reproductive strategy to host alternation by producing higher quality offspring (on average, 3·1% faster development and 2·1% larger size) without compromising offspring number. However, the former produced eggs that were slightly, albeit significantly, smaller when it fed on alternating hosts as compared with a preferred host. Moreover, host order affected reproductive performance in host generalists (e.g. 2·8% larger eggs when the first feeding was performed on a non-preferred host), but not in a host specialist. We conclude that immediate effects of environmental fluctuation on parasite fitness depend on the degree of host specialization.


Genetically contextual effects of smoking on genome wide DNA methylation.
Smoking is the leading cause of death in the United States. It exerts its effects by increasing susceptibility to a variety of complex disorders among those who smoke, and if pregnant, to their unborn children. In prior efforts to understand the epigenetic mechanisms through which this increased vulnerability is conveyed, a number of investigators have conducted genome wide methylation analyses. Unfortunately, secondary to methodological limitations, these studies were unable to examine methylation in gene regions with significant amounts of genetic variation. Using genome wide genetic and epigenetic data from the Framingham Heart Study, we re-examined the relationship of smoking status to genome wide methylation status. When only methylation status is considered, smoking was significantly associated with differential methylation in 310 genes that map to a variety of biological process and cellular differentiation pathways. However, when SNP effects on the magnitude of smoking associated methylation changes are also considered, cis and trans-interaction effects were noted at a total of 266 and 4353 genes with no marked enrichment for any biological pathways. Furthermore, the SNP variation participating in the significant interaction effects is enriched for loci previously associated with complex medical illnesses. The enlarged scope of the methylome shown to be affected by smoking may better explicate the mediational pathways linking smoking with a myriad of smoking related complex syndromes. Additionally, these results strongly suggest that combined epigenetic and genetic data analyses may be critical for a more complete understanding of the relationship between environmental variables, such as smoking, and pathophysiological outcomes.


Structural changes in the cytoplasmic domain of phospholamban by phosphorylation at Ser16: a molecular dynamics study.
Phospholamban is a 52-residue integral membrane protein that regulates the activity of the sarcoplasmic reticulum calcium pump in cardiac muscle. Its inhibitory action is relieved when phospholamban is phosphorylated at Ser16 by cAMP-dependent protein kinase. To computationally explore all possible conformations of the phosphorylated form, and thereby to understand the structural effects of phosphorylation, replica-exchange molecular dynamics (REMD) was applied to the cytoplasmic domain that includes Ser16. The simulations showed that (i) without phosphorylation, the region from Lys3 to Ser16 takes all alpha-helical conformations; (ii) when phosphorylated, the alpha-helix is partially unwound in the C-terminal part (from Ser10 to Ala15) resulting in less extended conformations; (iii) the phosphate at Ser16 forms salt bridges with Arg9, Arg13, and/or Arg14; and (iv) the salt bridges with Arg13 and Arg14 distort the alpha-helix and induce unwinding of the C-terminal part. We then applied conventional all-atom molecular dynamics simulations to the full-length phospholamban in the phospholipid bilayer. The results were consistent with those obtained with REMD simulations, suggesting that the transmembrane part of phospholamban and the lipid bilayer itself have only minor effects on the conformational changes in the cytoplasmic domain. The distortions caused by the salt bridges involving the phosphate at Ser16 readily explain the relief of the inhibitory effect of phospholamban by phosphorylation, as they will substantially reduce the population of all helical conformations, which are presumably required for the binding to the calcium pump. This will also be the mechanism for releasing the phosphorylated phospholamban from kinase.


MicroRNA-23a/24-2/27a as a potential diagnostic biomarker for cancer: A systematic review and meta-analysis.
An increasing number of studies have proven that microRNAs play an important role in the occurrence, development and prognosis of various types of cancer. As a vital gene cluster, the microRNA (miR)-23a/24-2/27a cluster may be an important marker for predicting cancer prognosis and tumor progression. A search was conducted through PubMed, Medline and the Cochrane Library to identify studies investigating the association between the miR-23a/24-2/27a cluster and cancer, and the identified related studies were included in the present meta-analysis. The strength of the association was assessed by hazard ratio (HR) and its 95% confidence interval (95% CI). A total of 21 studies were included in this meta-analysis. The results indicated that a high level of miR-23a exerted a significant effect on overall survival (OS) (HR=2.33, 95% CI: 1.18-4.58; P=0.014), but not on disease-free survival (DFS)/recurrence-free survival (RFS) (HR=1.13, 95% CI: 0.37-3.44; P=0.836). There was an obvious statistically significant association between OS and the expression of miR-24 (HR=2.49, 95% CI: 1.84-3.37; P=0.000), particularly in the digestive system (pooled HR=2.99, 95% CI: 2.17-4.13, P=0.000). In addition, the result suggested a statistically significant association between the expression of miR-27a and OS (pooled HR=1.89, 95% CI: 1.32-2.69; P=0.001), as well as DFS/RFS/progression-free survival (HR=2.19, 95% CI: 1.29-3.70; P=0.003), particularly in renal cell carcinoma (HR=2.30, 95% CI: 1.16-4.67; P=0.017). A subgroup analysis by ethnicity, cancer type and statistical methodology was performed. There was no obvious publication bias. In conclusion, the present study demonstrated that the miR-23a/24-2/27a cluster may be a useful marker for predicting cancer prognosis and tumor progression.


Structural and functional comparisons of the environmental microbiota of pond and tank environments at different locations for the commercial aquaculture of American shad.
The environmental microbiota plays a significant role in the growth and development of aquatic life. In recent years, American shad has become an important economic aquaculture species in China. However, information on the correlation between the growth of American shad and the aquaculture environment is limited. Through 16S rDNA-based sequencing, the microbiota communities in ponds at different locations (Jiangyin and Yancheng in Jiangsu, China) were investigated. The results showed that the richness and diversity of the microbiota in the pond were greater than those in the tank at the same location. At the phylum level, Proteobacteria and Firmicutes were more abundant in the samples from Jiangyin than in those from Yancheng. In addition, the ratio of Firmicutes to Bacteroidetes was larger in the JYT samples than in the YCT samples, which implied that the American shad cultured in the Jiangyin environment tended to be fatter than those cultured in Yancheng. The body weight data also confirmed this finding. Moreover, the proportions of functional annotations in the samples from the Jiangyin and Yancheng environments were similar, but there were differences between the overall levels. Our results highlighted the correlations between the environmental microbiome and the growth tendency of American shad.


The incidence of cystic fibrosis in Scotland calculated from heterozygote frequencies.
The incidence of cystic fibrosis (CF) has previously been calculated from epidemiological surveys and from neonatal screening. With the cloning of the CF gene it has become possible to derive incidence figures from heterozygote frequencies, provided that the distribution of mutant alleles among healthy carriers is the same as among affected people. We have estimated the allele frequencies for four CF mutations, AF508, G551D, G542X and R117H, in 14360 unselected women undergoing antenatal heterozygote screening. The proportion of R117H, an allele of known mild effect, was much greater for heterozygotes than for homozygotes. The incidence of CF was therefore calculated from the heterozygote frequencies of AF508, G551D and G542X in a larger cohort of 27 161 successively screened women. The point estimate for the incidence of CF in the Scottish population was 1 in 1984, with 95% confidence intervals of 1 in 1692 to 1 in 2336.


[Degree of satisfaction of patients continuing overactive bladder treatment with mirabegron].
Overactive bladder syndrome is chronic disease with high prevalence rate (9-42%). This syndrome requires long term therapy, but the treatment persistence is after 3 months over all 26% with further decline in one-year period as low as 18.5%. Main reasons for stopping the treatment are low efficacy, the medication didnt work as expected and side effects. How much satisfied are patients with mirabegron persisting on its treatment? To answer this question, we provided secondary analysis of multicentre follow-up study of patients on mirabegron. We compared subjective and objective parameters between patients continuing mirabegron treatment and those who discontinued the medication. Secondary analysis multicentre prospective follow-up. Ob/Gyn department First Faculty of Medicine, Charles University and General University Hospital, Prague. It is secondary analysis of multicentre prospective study following patients with mirabegron 50 mg treatment. We have analysed objective data from micturition diary and subjective data using visual analogue scales (UB-VAS - urgency bother visual analogue scale, and TS-VAS - treatment satisfaction visual analogue scale) and compared data between the group of patients continuing mirabegron treatment and patients who stopped the medication during the study. We included 206 patients (176 women, 30 men) with diagnosis of overactive bladder. Patients continuing the treatment (group n1) had baseline UB-VAS 70.1 vs. 75.0 (p = n.s.) in patients who stopped the medication during the follow-up period (group n2). Baseline episodes of severe urgency and urge incontinence where n1 - 5.1 vs. n2 - 6.2 (p = n.s.). Six months urgency bother score UB-VAS was n1 - 32.4 vs. n2 - 58.9 (p < 0,001). Treatment satisfaction TS-VAS was n1 - 80.3 vs. n2 - 57.7 (p < 0,001). Number of severe urgencies with or without urge incontinence was after 6 months n1 - 2.1 vs. n2 - 3.3 (p = n.s.), lower in group continuing the treatment. When comparing the data between patients stopping the medication for reason of low efficacy (group s1) with patients stopping for other reasons (group s2) UB-VAS bas: s1 - 68.5 vs. s2 - 43.9 (p = 0.001); TS VAS s1 - 45.1 vs. s2 - 58.4 (p = n.s.) and number of severe urgency with or without incontinence s1 - 5.9 vs. s2 - 3.2 (p = 0.009). Our data shows that patients expectation on treatment with mirabegron is not low. Patients accept treatment either without side effects or with decrease of severe urgency with or without urge incontinence around 50%. Regardless the reason the patients continuing the treatment scale treatment satisfaction - TS-VAS over 70 points.


[A method of rational particle counting (author's transl)].
A method is described, which simplifies the counting of particles in a known volume. The counting of neurons of a brain region is used as an example for this procedure. The total volume is processed by histological serial sections. The total area of all sections of this volume is divided into counting squares by an ocular grid, from which we get a sample. This sample is described by the frequency distribution. The real number of particles in the volume is estimated by the values of the frequency distribution. The way of dividing, the size and the way of selection of these counting squares for a certain exactness of the estimation is determined. The method can be used for non-systematic distribution of particles in morphological structures.


Morphological and functional alterations of the ductus arteriosus in a chicken model of hypoxia-induced fetal growth retardation.
The hypoxic conditions in which children with intrauterine growth retardation (IUGR) develop are hypothesized to alter the development of the ductus arteriosus (DA). We aimed to evaluate the effects of in ovo hypoxia on chicken DA morphometry and reactivity. Hypoxia (15% O2 from day 6 to 19 of the 21-d incubation period) produced a reduction in the body mass of the 19-d fetuses and a shortening of right and left DAs. However, ductal lumen and media cross-sectional areas were not affected by hypoxia. The ductal contractions induced by oxygen, KCl, H2O2, 4-aminopyridine, and endothelin-1 were similar in control and hypoxic fetuses. In contrast, the DAs from the hypoxic fetuses showed increased contractile responses to norepinephrine and phenylephrine and impaired relaxations to acetylcholine, sodium nitroprusside, and isoproterenol. The relaxations induced by 8-Br-cGMP, forskolin, Y-27632, and hydroxyfasudil were not altered by chronic hypoxia. In conclusion, chronic in ovo hypoxia-induced growth retardation in fetal chickens and altered the response of the DA to adrenergic agonists and to endothelium-dependent and -independent relaxing agents. Our observations support the concept that prolonged patency of the DA in infants with IUGR may be partially related with hypoxia-induced changes in local vascular mechanisms.


Morphologic changes in the rat enamel organ following a single intraperitoneal injection of sodium fluoride.
Enamel organs in developing teeth of young rats were studied after single intraperitoneal injections of a high dose of sodium fluoride (60 mg NaF/kg body wt.). The study employed primarily light microscopy, but electron microscopy was used to clarify some of the light microscopic findings. The pathogenesis of the fluoride-induced changes was followed during 72 h. Cellular changes were consistently found in the molars, but were never seen in the incisors. In the maxillary molars, ameloblastic injury was most commonly seen on the mesial surfaces of the cusps. One hour after injection, the most prominent findings were swollen mitochondria in the secretory ameloblasts and cleft formations between the ameloblasts and the enamel matrix. The clefts were filled with a stippled material. Some of the clefts gradually expanded to cystic cavities. The stippled material began to calcify after 24 h and formed small, darkly stained globules. After 72 h dearranged ameloblasts were found as islands intermingled with calcified rounded structures in the stellate reticulum. In stratum intermedium numerous atypic autophagic vacuoles appeared 2 h after injection. No light microscopic changes were observed in the postsecretory ameloblasts.


Microangiopathic hemolytic anemia associated with metastatic carcinoma of the colon.
We have reported a case of microangiopathic hemolytic anemia and metastatic carcinoma of the colon in a 38-year-old woman. Despite the mucin content and the high incidence of carcinoma of the colon in western populations, there are only four cases of colon cancer among the 77 reported cases of malignancy-associated MHA.


[Expression of protein p53: the marker of low neoplastic cell differentiation in thyroid carcinoma].
The aim of the study was to evaluate the frequency of positive staining of p53 protein in the course of thyroid carcinoma and to establish the prognostic value of such expression. Immunohistochemical staining of frozen sections was performed in 159 patients with thyroid carcinoma (139 females and 29 males). The average age was 50.4 years. All patients have been followed-up for at least 5 years and the average time of observation was 10.7 years. There were 46% patients with papillary carcinoma, 39% with follicular carcinoma and 15% with poorly differentiated thyroid carcinoma in the study group. Additionally, staining was performed in 6 cases of anaplastic carcinoma. The expression of p53 protein was assessed by two-degree scale: a medium (10% of cells with positive staining) and strong (50% of positive staining). In the group of 135 patients with well differentiated thyroid carcinoma the expression of p53 protein has been found in 55 cases (40.1%) but significantly more often in medium (69.1%) than in strong degree (30.9%). Significantly more often in expression has occurred among patients with poorly differentiated carcinoma (66%) and anaplastic carcinoma (100% of patients with strong expression in all cases). Comparing the frequency of p53 protein expression on every stage of disease, no significant difference has been found. The presence of p53 protein has not correlated with reduced changes in survival. In multivariate analyses the expression of p53 protein have not shown prognostic value. The expression of p53 protein has concerned the poorly differentiated carcinomas of thyroid and has not correlated with the stage of disease. No influence of expression on clinical course of the well differentiated thyroid carcinoma has been found. Worse prognoses have correlated with poor differentiated thyroid cancer cases and with strong expression of p53 protein.


Dynamics of supercooled and glassy dipropyleneglycol dibenzoate as functions of temperature and aging: Interpretation within the coupling model framework.
Dielectric relaxation measurements of a typical small molecular glassformer, dipropyleneglycol dibenzoate show the presence of two secondary relaxations. Their dynamic properties differ in the equilibrium liquid and glassy states, as well as the changes during structural recovery after rapid quenching the liquid to form a glass. These differences enable us to identify the slower secondary relaxation as the genuine Johari-Goldstein (JG) beta-relaxation, acting as the precursor of the primary alpha-relaxation. Agreement between the JG beta-relaxation time and the independent relaxation time of the coupling model leads to predicted quantitative relations between the JG beta-relaxation and the alpha-relaxation that are supported by the experimental data.


Muscle metabolites and performance during high-intensity, intermittent exercise.
Six men were studied during four 30-s "all-out" exercise bouts on an air-braked cycle ergometer. The first three exercise bouts were separated by 4 min of passive recovery; after the third bout, subjects rested for 4 min, exercised for 30 min at 30-35% peak O2 consumption, and rested for a further 60 min before completing the fourth exercise bout. Peak power and total work were reduced (P < 0. 05) during bout 3 [765 +/- 60 (SE) W; 15.8 +/- 1.0 kJ] compared with bout 1 (1,168 +/- 55 W, 23.8 +/- 1.2 kJ), but no difference in exercise performance was observed between bouts 1 and 4 (1,094 +/- 64 W, 23.2 +/- 1.4 kJ). Before bout 3, muscle ATP, creatine phosphate (CP), glycogen, pH, and sarcoplasmic reticulum (SR) Ca2+ uptake were reduced, while muscle lactate and inosine 5'-monophosphate were increased. Muscle ATP and glycogen before bout 4 remained lower than values before bout 1 (P < 0.05), but there were no differences in muscle inosine 5'-monophosphate, lactate, pH, and SR Ca2+ uptake. Muscle CP levels before bout 4 had increased above resting levels. Consistent with the decline in muscle ATP were increases in hypoxanthine and inosine before bouts 3 and 4. The decline in exercise performance does not appear to be related to a reduction in muscle glycogen. Instead, it may be caused by reduced CP availability, increased H+ concentration, impairment in SR function, or some other fatigue-inducing agent.


Bacillomycin D Produced by Bacillus amyloliquefaciens Is Involved in the Antagonistic Interaction with the Plant-Pathogenic Fungus Fusarium graminearum.
Fusarium graminearum (teleomorph: Ascomycota, Hypocreales, Gibberella, Gibberella zeae) is a destructive fungal pathogen that threatens the production and quality of wheat and barley worldwide. Controlling this toxin-producing pathogen is a significant challenge. In the present study, the commercially available strain Bacillus amyloliquefaciens (Bacteria, Firmicutes, Bacillales, Bacillus) FZB42 showed strong activity against F. graminearum The lipopeptide bacillomycin D, produced by FZB42, was shown to contribute to the antifungal activity. Purified bacillomycin D showed strong activity against F. graminearum, and its 50% effective concentration was determined to be approximately 30 μg/ml. Analyses using scanning and transmission electron microscopy revealed that bacillomycin D caused morphological changes in the plasma membranes and cell walls of F. graminearum hyphae and conidia. Fluorescence microscopy combined with different dyes showed that bacillomycin D induced the accumulation of reactive oxygen species and caused cell death in F. graminearum hyphae and conidia. F. graminearum secondary metabolism also responded to bacillomycin D challenge, by increasing the production of deoxynivalenol. Biological control experiments demonstrated that bacillomycin D exerted good control of F. graminearum on corn silks, wheat seedlings, and wheat heads. In response to bacillomycin D, F. graminearum genes involved in scavenging reactive oxygen species were downregulated, whereas genes involved in the synthesis of deoxynivalenol were upregulated. Phosphorylation of MGV1 and HOG1, the mitogen-activated protein kinases of F. graminearum, was increased in response to bacillomycin D. Taken together, these findings reveal the mechanism of the antifungal action of bacillomycin D.IMPORTANCE Biological control of plant disease caused by Fusarium graminearum is desirable. Bacillus amyloliquefaciens FZB42 is a representative of the biocontrol bacterial strains. In this work, the lipopeptide bacillomycin D, produced by FZB42, showed strong fungicidal activity against F. graminearum Bacillomycin D caused morphological changes in the plasma membrane and cell wall of F. graminearum, induced accumulation of reactive oxygen species, and ultimately caused cell death in F. graminearum Interestingly, when F. graminearum was challenged with bacillomycin D, the deoxynivalenol production, gene expression, mitogen-activated protein kinase phosphorylation, and pathogenicity of F. graminearum were significantly altered. These findings clarified the mechanisms of the activity of bacillomycin D against F. graminearum and highlighted the potential of B. amyloliquefaciens FZB42 as a biocontrol agent against F. graminearum.


A Saccharomyces cerevisiae gene encoding a potential adenine phosphoribosyltransferase.
The nucleotide sequence of a Saccharomyces cerevisiae gene encoding a potential adenine phosphoribosyltransferase (APRT) has been determined. The protein encoded by this gene shows a high degree of similarity with APRTs from a variety of other species. The S. cerevisiae gene, named APT2, has been mapped to chromosome IV. The sequence has been deposited in the GenBank data library under Accession Number L14434.


Kinetic studies and bioactivity of potential manzamine prodrugs.
The manzamines represent a class of marine natural products that show considerable promise in the control of malaria but generate GI distress in rodents when administered orally in high doses. In an effort to generate manzamine prodrugs with improved antimalarial activity and reduced GI toxicity, we prepared acetylated 8-hydroxymanzamine A analogues including 8-acetoxymanzamine A (3) and 8,12-diacetoxymanzamine A (4), and 8-methoxymanzamine A (5) beginning with 8-hydroxymanzamine A (2). The semisynthetic analogues were assayed for antimalarial and antimicrobial activities, cytotoxicity, and biological and chemical stability. Due to gradual hydrolysis of the ester group, application of monoacetate 3 as an antimalarial prodrug was investigated. The in vitro and in vivo bioassays show that acetylated analogues exhibit significant antimalarial activity (IC50( 3) 9.6-30 ng/mL), which are comparable to the parent molecule; however the monoaceate 3 was shown to actually produce higher toxicity at 30 mg/kg when administered orally.


Lymphohematopoietic malignancies and oil exploitation in Koprivnica-Krizevci County, Croatia.
To determine lymphohematopoietic malignancy incidence rates between populations living near oil-gas fields (two regions) compared with those living in areas free from oil-gas exploitation (one region) in Koprivnica-Krizevci County, Croatia, 803 new cases of lymphohematopoietic malignancies were studied for two time periods: 1971-1980 (prior to oil-gas exploitation) and 1981-2000 (during oil-gas exploitation). The results showed considerable geographic differences in the incidences of chronic myeloid leukemia and multiple myeloma. The authors conclude that proximity to oil or gas fields represents an increased population risk of developing lymphohematopoietic cancers.


Immunologic assessment determined by response to IL-2 and immunophenotyping of tumor-infiltrating lymphocytes, of invaded and non invaded lymph nodes and of peripheral blood lymphocytes from twenty-one patients with primary laryngeal cancer.
The aim of the study was to identify the lymphocyte sets and/or subsets possibly involved in the response to malignant cells. For this purpose we have investigated both the cells at the tumor site, i.e. tumor-infiltrating lymphocytes (TIL), and the cells present in the draining lymph nodes, either invaded or non invaded, as well as the peripheral blood lymphocytes from twenty-one patients with primary laryngeal epidermoid carcinoma. The functional assay was carried out by the proliferative response to mitogens, to Interleukin 2 and to their association, the surface immunophenotyping was performed with a large panel of monoclonal antibodies. TIL are the most responsive cells to mitogens, while the responsiveness of TIL and of LN cells to IL-2 was in the same range. PHA-activated TIL are the most responsive cells to IL-2. Our data indicate that TIL do show in vitro, and probably also in vivo, "activation" with elevated responsiveness to IL-2. The surface phenotype showed a strikingly increased proportion of T8+ cells in TIL as compared to T8+ cells in all types of LN, thus confirming within TIL variable, but high, proportions of clones which display cytolytic activity, possibly induced by IL-2. Our data seem to support the perspective for a therapeutic approach in vivo with IL-2, which via its influence on TIL, may act on tumor cells.


Annular Elastolytic Giant Cell Granuloma Successfully Treated With Adalimumab Subsequently Complicated by Drug-Induced Lupus.
We report a 51-year-old female with a 3-year history of recalcitrant annular elastolytic giant cell granuloma (AEGCG) who was effectively treated with the anti-tumor necrosis factor (TNF)-alpha antibody, adalimumab. Her disease was refractory to topical glucocorticoids, intralesional glucocorticoids, narrow-band ultraviolet light (UV)-B phototherapy and cyclosporine. During her treatment with adalimumab she developed a positive anti-nuclear-antibody and double-stranded-DNA antibody and her treatment was terminated. Our findings suggest that adalimumab is an efficacious therapeutic alternative for the treatment of annular elastolytic giant cell granuloma unresponsive to standard therapies, however drug-induced lupus is a potential side effect that clinicians must be cognizant of. To our knowledge, this is the first time adalimumab has successfully been used in the treatment of AEGCG.
J Drugs Dermatol. 2017;16(2):169-171.
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[Effects of telmisartan on inflammation and fibrosis after acute myocardial infarction in rats].
To explore the protection mechanisms of telmisartan on inflammation and fibrosis after myocardial infarction in rats. The model of acute myocardial infarction (AMI) was established by ligating left anterior descending coronary artery. The surviving rats were divided into AMI (AMI) and telmisartan treatment (telmisartan) groups. And another sham operation group (sham) was designated (n = 8). At the end of study, total heart weight (THW), left ventricular weight (LVW) and weight index were measured; myocardial infarction and inflammatory reactions detected by hematoxylin and eosin and Masson staining; the serum levels of C-reactive protein (CRP), tumor necrosis factor-alpha (TNFα), monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6) and interleukin-1 beta (IL-1β) by enzyme-linked immunosorbent assay (ELISA); the levels of transforming growth factor 1 (TGFβ1), collagen I, collagen III and MMP9 mRNA in myocardial tissue by reverse transcription-polymerase chain reaction (RT-PCR); the expressions of TGFβ1, collagen I, collagen III, matrix metallopeptidase 9 (MMP9) and nuclear factor-kappa B (NF-κB) by Western blot. Compared with sham group, significant pathological changes of myocardium occurred in AMI group. The serum levels of CRP [(472 ± 132) vs (104 ± 28) ng/ml], TNFα [(229 ± 41) vs (18 ± 5) pg/ml], MCP-1[(558 ± 116) vs (158 ± 20) pg/ml], IL-6 [(404 ± 63) vs (21 ± 4) pg/ml] and IL-1β [(625 ± 145) vs (189 ± 34) pg/ml] increased (P < 0.05). RT-PCR analysis showed that the expression levels of TGFβ1, collagen I, collagen III and MMP9 increased significantly. The results of Western blot were consistent and NF-κB was activated significantly (P < 0.05). Compared with AMI group, the above-mentioned indicators decreased obviously in telmisartan group (P < 0.05). Telmisartan may regulate inflammation and myocardial fibrosis after acute myocardial infarction by signaling pathways of NF-κB and TGFβ in rats.


[Location of cochlear defect by AR spectrum analysis of TEOAE].
A new method is proposed in this paper to locate the position of hearing loss (high frequency, middle frequency or low frequency). The method is based on AR spectrum analysis of the transient evoked oto acoustic emission (TEOAE). The results obtained are essentially the same as those obtained by DPGRAM, which comes from DPOAE. Thus, a new field of TEOAE application is found by the proposed method.


4-Bromodiphenyl Ether Induces Germ Cell Apoptosis by Induction of ROS and DNA Damage in Caenorhabditis elegans.
Polybrominated diphenyl ethers may affect male reproductive function; however, the underlying mechanism is still uncertain. By using Caenorhabditis elegans, a commonly used model to study basic biological processes in apoptosis, we investigated the toxic effects of 4-bromodiphenyl ether (BDE-3), the most fundamental mono-BDE generated from degradation of polybrominated diphenyl ethers in the environment. We found that BDE-3 treated worms exhibited decreased life spans, impaired fecundity and delayed egg laying. BDE-3 induced dose-dependent germ cell apoptosis in wild-type N2 strain; however, this effect was blocked in mutants of p53/cep-1 and DNA damage response gene hus-1. Moreover, the knockout of the MAPK kinases (mutants mek-1 and sek-1) and the p53 antagonist protein ABL-1 (abl-1), which are essential for stress-induced germ cell apoptosis, also abrogated the germ cell apoptosis induced by BDE-3. Generation of reactive oxygen species (ROS) in intact animals was determined by a fluorescent probe, 2,7-dichlorofluorescein diacetate, and the ROS level was significantly elevated by BDE-3 treatment. Microarray analysis on gene expression profiles further revealed the possible pathways involved in BDE-3 toxicity. Overall, our findings suggested that BDE-3 could induce reproductive dysfunction and germ cell apoptosis in C. elegans by induction of ROS and DNA damage.


Duodenal mucosa FOXP3 expression in different etiologies of lymphocytic duodenosis.
In celiac disease there is an increase of lymphocytes expressing FOXP3 in the intestinal mucosa associated with varying degrees of villous atrophy. Our aim was to evaluate FOXP3 expression in duodenal mucosa with lymphocytic enteritis according to aetiology and correlation with lymphocytes T-γδ. We compared three adult patient groups suffering lymphocytic enteritis: celiacs following a gluten-free diet (n=12), first-degree relatives of celiac patients with genetic risks (n=14) and patients with functional dyspepsia (n=14), along with a control group not suffering from duodenal enteritis (n=16). The population of duodenal lymphocytes was analysed by immunohistochemistry assays for CD3+ characterisation and FOXP3 expression. Quantification of lymphocytes T-γδ in duodenal mucosa was performed by flow cytometry in fresh tissue samples. Presence of lymphocytes T-γδ was significantly higher in the group of celiac individuals compared to the group of relatives of these individuals (37.44 vs 5,52: p<0.0001) and the group with functional dyspepsia (37.44 vs 11.76: p=0.008). FOXP3 expression was also significantly higher in the celiac group than in the groups of relatives (18.85 vs 6.31; p=0.001) and functional dyspepsia patients (18.85 vs 7.61; p=0.023). The proportion of lymphocytes T-γδ and FOXP3- expressing lymphocytes was similar in the control group to that in the relatives or functional dyspepsia groups. Lymphocytic enteritis associated to celiac disease shows an increase of FOXP3 expression and lymphocytes T-γδ that is not detected in other etiologies of enteritis.


[Theoretical background of our original single-incision laparoscopic-assisted surgery technique for colon cancer via a periumbilical approach].
We assessed the theoretical background of our original single-incision laparoscopic-assisted surgery( SILS) technique involving a periumbilical approach. The subjects included 10 cases who underwent periumbilical SILS colectomy and had their surgical wounds photographed before and after skin incision between September 2009 and October 2010. Using an image analyzer, we estimated the theoretical oval area after a 3/4-circumferential periumbilical skin incision, the actual oval area after placement of the wound retractor, and the length of the skin incision. The mean oval area after the placement of the wound retractor was 2.9 times (range: 1.6-5.0 times) larger than that of the theoretical area. The square of the length of the skin incision positively correlated with the actual oval area created by placing the wound retractor( p=0.04, r=0.67). There were 5 patients, whose actual oval area was ≤700 mm2, and thus required additional radial skin incision( s)( 1 in 3 cases, 2 in 1 case and 3 in 1 case). When performing our original SILS via the periumbilical approach, the area of the actual surgical window can be predicted by measuring the distance from the center of the umbilicus to its edge.


Basal ganglia calcification in a patient with beta-propeller protein-associated neurodegeneration.
Beta-propeller protein-associated neurodegeneration is a newly described X-linked dominant condition due to heterozygous mutations in WDR45. The condition is associated with characteristic changes on brain magnetic resonance imaging. Previous literature relating to this disorder has not specifically referred to intracranial calcification. A female patient presented with significant developmental delay in early childhood and subsequently demonstrated neurodegeneration with progressive dystonia and dementia in her third decade. Brain magnetic resonance imaging revealed low signal in the substantia nigra and both globus pallidi on T2-weighted imaging, with no eye-of-the-tiger sign. Computed tomography revealed bilateral dense calcification of the globus pallidus. We performed Sanger sequencing of the WDR45 gene in the patient and her parents. We identified a heterozygous c.488del C p.Pro163Argfs*34 variant in exon 8 of WDR45. Neither parent carried the same mutation, indicating that the molecular change had occurred de novo. Although the characteristic features of beta-propeller protein-associated neurodegeneration were present in our patient, the observation of basal ganglia calcification was considered atypical. Previous descriptions of basal ganglia calcification in individuals with neuronal brain iron accumulation led us to review the frequency of calcification in these disorders.


Isolation and seasonal effects on characteristics of fulvic acid isolated from an Australian floodplain river and billabong.
Fulvic acids from an Australian floodplain river and billabong were isolated using DEAE and DAX-8 resins, and characterised with the use of size exclusion chromatography and solid-state CP-MAS (13)C NMR spectroscopy. Differences between the two resin isolates were evident. Fulvic acids isolated using DEAE-cellulose had higher apparent M(n) and M(w) values, while the DAX-8 resin showed a slight preference for aliphatic components. Fulvic acids from the river and billabong had the same functional groups present, however, the river fulvic acids had higher apparent M(n) (number average molecular weight) and M(w) values (weight average molecular weight), and were more polydisperse than the billabong fulvic acid. There were no significant changes in the characteristics of the fulvic acid isolated from the river at four sampling times: summer, autumn, winter and spring. In contrast, fulvic acids isolated from a billabong displayed seasonal variation in molecular weights. This work emphasizes the importance in ecological studies of isolation procedure for the operationally defined fulvic acids.


Theoretical and experimental studies of enflurane. Infrared spectra in solution, in low-temperature argon matrix and blue shifts resulting from dimerization.
Theoretical studies are performed on enflurane (CHFCl-CF(2)-O-CHF(2)) to investigate the conformational properties and vibrational spectra. Calculations are carried out at the B3LYP/6-31G(d) level along with a natural bond orbital (NBO) analysis. Experimental infrared spectra are investigated in carbon tetrachloride solution at room temperature and in argon matrix at 12 K. In agreement with previously reported data (Pfeiffer, A.; Mack, H.-G.; Oberhammer, H. J. Am. Chem. Soc. 1998, 120, 6384), it is shown that the four most stable conformers possess a trans configuration of the C-C-O-C skeleton and a gauche orientation of the CHF(2) group (with respect to the central C-O bond). These conformations are favored by electrostatic interaction between the H atom of the CHF(2) group and the F atoms of the central CF(2) group. Hyperconjugation effects from the O lone pairs to the antibonding orbitals of the neighboring C-H and C-F bonds also contribute to the stability of the four conformers. The vibrational frequencies, infrared intensities, and potential energy distributions are calculated at the same level of theory for the most stable conformers. On the basis of the theoretical results, these conformers are identified in an argon matrix. The influence of the concentration on the nu(CH) vibrations suggests the formations of higher aggregates in solution. Theoretical calculations are carried out on the enflurane dimer. The results show that the dimer is formed between two enflurane conformers having the largest stability. The dimer has an asymmetric cyclic structure, the two enflurane molecules being held together by two nonequivalent C-H...F hydrogen bonds, the C-H bond of the CHFCl group acting as a proton donor, and one of the F atoms of the CHF(2) groups acting as a proton acceptor. The theory predicts a contraction of 0.0014-0.0025 A of the two CH bonds involved in the interaction along with a blue shift of 30-38 cm(-1) of the corresponding nu(C-H) bands, in good agreement with the blue shifts of 35-39 cm(-1) observed in an argon matrix.


Direct digital radiography versus storage phosphor radiography in the detection of wrist fractures.
To define the value of digital radiography with a clinical flat panel detector system for evaluation of wrist fractures in comparison with state of the art storage phosphor radiography. Hard copy images of 26 fractured wrist specimens were acquired with the same exposure dose on a state of the art storage phosphor radiography system and a clinical flat panel detector. Image features like cortical bone surface, trabecular bone, soft tissues and fracture delineation were independently analysed by 4 observers using a standardised protocol. Image quality ratings were evaluated with an analysis of variance (ANOVA). Flat panel detector radiographs were rated superior with respect to cortical and trabecular bone representation as well as fracture evaluation, while storage phosphor radiographs produced better soft tissue detail. In some of the observed image quality aspects, the performance of caesium iodide/amorphous silicon flat panel detector exceeds state of the art storage phosphor radiography. This makes it well suited for skeletal imaging particularly in trauma as seen in the detection of wrist fractures.


Adiponectin and AdipoR1 regulate PGC-1alpha and mitochondria by Ca(2+) and AMPK/SIRT1.
Adiponectin is an anti-diabetic adipokine. Its receptors possess a seven-transmembrane topology with the amino terminus located intracellularly, which is the opposite of G-protein-coupled receptors. Here we provide evidence that adiponectin induces extracellular Ca(2+) influx by adiponectin receptor 1 (AdipoR1), which was necessary for subsequent activation of Ca(2+)/calmodulin-dependent protein kinase kinase beta (CaMKKbeta), AMPK and SIRT1, increased expression and decreased acetylation of peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1alpha), and increased mitochondria in myocytes. Moreover, muscle-specific disruption of AdipoR1 suppressed the adiponectin-mediated increase in intracellular Ca(2+) concentration, and decreased the activation of CaMKK, AMPK and SIRT1 by adiponectin. Suppression of AdipoR1 also resulted in decreased PGC-1alpha expression and deacetylation, decreased mitochondrial content and enzymes, decreased oxidative type I myofibres, and decreased oxidative stress-detoxifying enzymes in skeletal muscle, which were associated with insulin resistance and decreased exercise endurance. Decreased levels of adiponectin and AdipoR1 in obesity may have causal roles in mitochondrial dysfunction and insulin resistance seen in diabetes.