Transcription Factor PecS Mediates Agrobacterium fabrum Fitness and Survival.
The transcriptional regulator PecS is encoded by select bacterial pathogens. For instance, in the plant pathogen Dickeya dadantii, PecS controls a range of virulence genes, including pectinase genes and the divergently oriented gene pecM, which encodes an efflux pump through which the antioxidant indigoidine is exported. In the plant pathogen Agrobacterium fabrum (formerly named Agrobacterium tumefaciens), the pecS-pecM locus is conserved. Using a strain of A. fabrum in which pecS has been disrupted, we show here that PecS controls a range of phenotypes that are associated with bacterial fitness. PecS represses flagellar motility and chemotaxis, which are processes that are important for A. fabrum to reach plant wound sites. Biofilm formation and microaerobic survival are reduced in the pecS disruption strain, whereas the production of acyl homoserine lactone (AHL) and resistance to reactive oxygen species (ROS) are increased when pecS is disrupted. AHL production and resistance to ROS are expected to be particularly relevant in the host environment. We also show that PecS does not participate in the induction of vir genes. The inducing ligands for PecS, urate, and xanthine, may be found in the rhizosphere, and they accumulate within the plant host upon infection. Therefore, our data suggest that PecS mediates A. fabrum fitness during its transition from the rhizosphere to the host plant. IMPORTANCE PecS is a transcription factor that is conserved in several pathogenic bacteria, where it regulates virulence genes. The plant pathogen Agrobacterium fabrum is important not only for its induction of crown galls in susceptible plants but also for its role as a tool in the genetic manipulation of host plants. We show here that A. fabrum PecS controls a range of phenotypes, which would confer the bacteria an advantage while transitioning from the rhizosphere to the host plant. This includes the production of signaling molecules, which are critical for the propagation of the tumor-inducing plasmid. A more complete understanding of the infection process may inform approaches by which to treat infections as well as to facilitate the transformation of recalcitrant plant species.


Using 100- instead of 120-kVp computed tomography to diagnose pulmonary embolism almost halves the radiation dose with preserved diagnostic quality.
Concern has been raised regarding the mounting collective radiation doses from computed tomography (CT), increasing the risk of radiation-induced cancers in exposed populations. To compare radiation dose and image quality in a chest phantom and in patients for the diagnosis of pulmonary embolism (PE) at 100 and 120 peak kilovoltage (kVp) using 16-multichannel detector computed tomography (MDCT). A 20-ml syringe containing 12 mg I/ml was scanned in a chest phantom at 100/120 kVp and 25 milliampere seconds (mAs). Consecutive patients underwent 100 kVp (n = 50) and 120 kVp (n = 50) 16-MDCT using a "quality reference" effective mAs of 100, 300 mg I/kg, and a 12-s injection duration. Attenuation (CT number), image noise (1 standard deviation), and contrast-to-noise ratio (CNR; fresh clot = 70 HU) of the contrast medium syringe and pulmonary arteries were evaluated on 3-mm-thick slices. Subjective image quality was assessed. Computed tomography dose index (CTDI(vol)) and dose-length product (DLP) were presented by the CT software, and effective dose was estimated. Mean values in the chest phantom and patients changed as follows when X-ray tube potential decreased from 120 to 100 kVp: attenuation +23% and +40%, noise +38% and +48%, CNR -6% and 0%, and CTDI(vol) -38% and -40%, respectively. Mean DLP and effective dose in the patients decreased by 42% and 45%, respectively. Subjective image quality was excellent or adequate in 49/48 patients at 100/120 kVp. No patient with a negative CT had any thromboembolism diagnosed during 3-month follow-up. By reducing X-ray tube potential from 120 to 100 kVp, while keeping all other scanning parameters unchanged, the radiation dose to the patient may be almost halved without deterioration of diagnostic quality, which may be of particular benefit in young individuals.


Mechanisms and physiological significance of the transport of the glutathione conjugate of 4-hydroxynonenal in human lens epithelial cells.
To study the mechanism and physiological significance of the transport of the glutathione (GSH) conjugate of 4-hydroxynonenal (4-HNE) from lens epithelial cells. HLE B-3 cells were treated with [(3)H] 4-HNE and efflux of its GSH conjugate, [(3)H] GS-HNE, into the medium was quantitated and characterized by HPLC and mass spectrometry. Inside-out vesicles (IOVs) were prepared from HLE B-3 cell membranes. The kinetics of adenosine triphosphate (ATP)-dependent uptake of GS-HNE and dinitrophenyl S-glutathione (DNP-SG) by these IOVs and inhibition of GS-HNE uptake by anti-RLIP76 IgG was studied. Localization of RLIP76 was studied by immunogold electron microscopy and kinetics of the adenosine triphosphatase (ATPase) activity of purified RLIP76 was determined. 4-HNE-induced apoptosis was compared in HLE-B3 cells coated with anti-RLIP76 IgG or preimmune IgG, by caspase activation assay. The results showed the presence of RLIP76 in plasma membranes of HLE B-3 cells and that it mediated ATP-dependent transport of GS-HNE as well as of DNP-SG. The transport was saturable with respect to GS-HNE (K(m) = 8.4 micro M), DNP-SG (100 micro M) as well as to ATP (K(m) 0.45 mM) and was sensitive to temperature and osmolarity of the medium. Anti-RLIP76 IgG inhibited approximately 65% of the transport of GS-HNE and DNP-SG, indicating that most of the transport was mediated by RLIP76. Compatible with its transport function, the EGTA- and ouabain-insensitive ATPase activity of purified RLIP76 was stimulated by DNP-SG and GS-HNE. 4-HNE-induced caspase activation in HLE-B3 cells was exacerbated when the transport of GS-HNE from these cells was blocked by anti-RLIP76 IgG. RLIP76 provides a defense against the deleterious effects of 4-HNE by transporting GS-HNE and can modulate apoptotic signaling by regulating the intracellular concentrations of 4-HNE.


Neural interactions mediating the detection of motion in the retina of the tiger salamander.
The neural circuitry underlying movement detection was inferred from studies of amacrine cells under whole-cell patch clamp in retinal slices. Cells were identified by Lucifer yellow staining. Synaptic inputs were driven by "puffing" transmitter substances at the dendrites of presynaptic cells. Spatial sensitivity profiles for amacrine cells were measured by puffing transmitter substances along the lateral spread of their processes. Synaptic pathways were separated and identified with appropriate pre- and postsynaptic pharmacological blocking agents. Two distinct amacrine cell types were found: one with narrow spread of processes that received sustained excitatory synaptic current, the other with very wide spread of processes that received transient excitatory synaptic currents. The transient currents found only in the wide-field amacrine cell were formed presynaptically at GABAB receptors. They could be blocked with baclofen, a GABAB agonist, and their time course was extended by AVA, a GABAB antagonist. Baclofen and AVA had no direct affect upon the wide-field amacrine cell, but picrotoxin blocked a separate, direct GABA input to this cell. The narrow-field amacrine cell was shown to be GABAergic by counterstaining with anti-GABA antiserum after it was filled with Lucifer yellow. Its narrow, spatial profile and sustained synaptic input are properties that closely match those of the GABAergic antagonistic signal that forms transient activity (described above), suggesting that the narrow-field amacrine cell itself is the source of the GABAergic interaction mediating transient activity in the inner plexiform layer (IPL). Other work has shown a GABAB sensitivity at some bipolar terminals, suggesting a population of bipolars as the probable site of interaction mediating transient action. The results suggest that two local populations of amacrine cell types (sustained and transient) interact with the two populations of bipolar cell types (transient forming and nontransient forming). These interactions underlie the formation of the change-detecting subunits. We suggest that local populations of these subunits converge to form the receptive fields of movement-detecting ganglion cells.


[Practical quality assurance in x-ray diagnosis (author's transl)].
The reproduction parameters of an radiographic are presented by those details of the various visualised objects which are important for diagnosis. These parameters determine the required technical quality. The following factors are suitable for assessing the technical quality of the radiographs: medium optical density, contrast, resolution, peak voltage, exposure time and exposure dose. The physician himself is expected to maintain a close watch on the quality of his x-ray examinations. The following measures are conductive to this end: 1. comparison of the current patient radiographs with the standard x-ray visualisations as seen in routine practice 2. control of the processing technique with test films, and 3. checking on the proper functioning of the x-ray equipment by means of a small phantom. Over and above this, the x-ray equipment should be checked for its efficiency status in yearly intervals by means of measurements effected with a standard phantom by an external institution.


Monitoring vaccine safety using the Vaccine Safety Datalink: utilizing immunization registries for pandemic influenza.
Mass vaccination campaigns during which new vaccines may be administered to many millions of people in a short period of time call for timely and accurate post-licensure surveillance to monitor vaccine safety. To address the need for timely H1N1 influenza vaccine safety information during the 2009-2010 H1N1 influenza pandemic, the Vaccine Safety Datalink (VSD) project assessed the feasibility and potential mechanisms for utilizing data from state and local immunization registries to capture vaccinations that would not otherwise be captured by the data systems of the participating VSD managed care organizations (MCOs). Three of the eight VSD sites were able to capture H1N1 immunization data electronically from the state and local registries, and one site was able to capture the immunizations through a paper-based system; however, the remaining four sites encountered various obstacles that prevented capture of such data. Additional work will be required at these sites to overcome the barriers, which included privacy and confidentiality laws, time constraints brought on by the pandemic, as well as data quality concerns.


Function of pituitary-thyroid axis in copper-deficient rats.
Thirty male weanling Sprague-Dawley rats were fed either a copper deficient diet (0.85 mg Cu/kg) or a copper-adequate diet (8 mg Cu/kg). After 7 weeks, the rats were fasted for 12 hours, and injected intravenously with thyrotropin-releasing hormone (TRH, 30 ng/100 g body weight). Six rats from each treatment were killed at 0, 0.5, 1, 2 and 4 hours after TRH injection. Sera obtained at 0, 0.5 and 1 hour, and at 0.2 and 4 hours were used for the radioimmunoassay of thyroid-stimulating hormone (TSH) and thyroxine (T4), respectively. Reduction in liver copper content confirmed that the rats fed the test diet were copper-deficient. Serum TSH levels appeared to have peaked at 30 minutes but declined to a level higher than basal at 1 hour. No difference in TSH response was observed between the 2 treatments. Serum T4 response to TRH was reduced in the copper-deficient as compared to the adequate rats at all time intervals. After 2 hours a slight elevation was observed in the controls, but marked elevations in T4 were observed in both treatments at 4 hours. This reduction in T4 levels could be due to an impaired T4 synthesis or release in copper-deficient rats.


Varicella-zoster virus (chickenpox) infection in pregnancy.
Congenital varicella syndrome, maternal varicella-zoster virus pneumonia and neonatal varicella infection are associated with serious fetomaternal morbidity and, not infrequently, mortality. Vaccination against varicella-zoster virus can prevent the disease, and outbreak control limits the exposure of pregnant women to the infectious agent. Maternal varicella-zoster immunoglobulin administration before rash development, with or without antiviral medication, can modify the progression of the disease.


Evidence of Rift Valley fever seroprevalence in the Sahrawi semi-nomadic pastoralist system, Western Sahara.
The increasing global importance of Rift Valley fever (RVF) is clearly demonstrated by its geographical expansion. The presence of a wide range of host and vector species, and the epidemiological characteristics of RVF, have led to concerns that epidemics will continue to occur in previously unaffected regions of Africa. The proximity of the Sahrawi territories of Western Sahara to endemic countries, such as Mauritania, Senegal, and Mali with periodic isolation of virus and serological evidence of RVF, and the intensive livestock trade in the region results in a serious risk of RVF spread in the Sahrawi territories, and potentially from there to the Maghreb and beyond. A sero-epidemiological survey was conducted in the Saharawi territories between March and April 2008 to investigate the possible presence of the RVF virus (RVFV) and associated risk factors. A two-stage cluster sampling design was used, incorporating 23 sampling sites. A total of 982 serum samples was collected from 461 sheep, 463 goats and 58 camels. Eleven samples (0.97%) tested positive for IgG against the RVFV. There were clusters of high seroprevalence located mostly in the Tifariti (7.69%) and Mehaires (7.14%) regions, with the Tifariti event having been found in one single flock (4/26 positive animals). Goats and older animals were at a significantly increased risk being seropositive (p = 0.007 and p = 0.007, respectively). The results suggest potential RVF activity in the study area, where intense livestock movement and trade with neighbouring countries might be considered as a primary determinant in the spread of the disease. The importance of a continuous field investigation is reinforced, in light of the risk of RVF expansion to historically unaffected regions of Africa.


Community professionals' compliance with consultants' recommendations for developmentally disabled children.
Much of the responsibility for managing children with developmental disabilities lies with professionals working in community services. In many instances advice and consultation for these frontline professionals is available from a regional consultation facility. The purpose of this study was to prospectively examine the relationship between community professional clients' perceptions of the quality of the consultations received and their subsequent compliance with the recommendations made to them for the management of children in their care. Two hundred and ninety-nine consultation recommendations were followed: 2 weeks after the advice had been provided, clients completed a general satisfaction questionnaire and a client perceptions questionnaire. Two months later, compliance with the recommendation was determined by a validated self-report questionnaire. Overall, compliance was good for 73% of the recommendations. Negative client perceptions regarding several aspects of the consultants' skills in conducting the consultation were strongly associated with subsequent noncompliance. The findings suggest that community professional compliance may be enhanced by attention to improving the process of consultation.


Converting an array of edge dislocations into a multi-vortex beam.
We theoretically show how, using a cylindrical lens, a Gaussian beam with a finite number of parallel zero-intensity lines (edge dislocations) is transformed into a vortex beam that carries orbital angular momentum (OAM) and topological charge (TC). Remarkably, while the original beam is assumed to carry a non-zero OAM and have no TC, the latter is shown to appear during free-space propagation. Considering two parallel center-symmetric zero-intensity lines located as an example, we look into the dynamics of generating two intensity nulls at the double focal length: with increasing distance between the vertical zero-intensity lines, two optical vortices are first generated on the horizontal axis before converging at the origin and then diverging along the vertical axis. Irrespective of the between-line distance, such an optical vortex has ${\rm TC} = - {2}$ at any distance from the optical axis, except for the original plane. With changing distance between the zero-intensity lines, the OAM that the beam carries is changing, taking positive and negative values, or a zero value at a certain between-line distance. We also show that if the number of zero-intensity lines is infinite, a vortex beam with finite OAM and infinite TC is generated.


Sonographic evaluation of visceral and subcutaneous fat in obese children.
To evaluate sonographic measurements of visceral and subcutaneous fat in children, and to investigate the usefulness of preperitoneal fat (PF) and the abdominal wall fat index (AFI) as parameters to determine visceral fat and presence of nonalcoholic fatty liver disease (NAFLD) in obese children. A case-control study of a sample including 44 children (22 cases and 22 controls) matched by sex and age. The Student t test and the Fisher exact test were utilized in the descriptive and bivariate analysis. The sonographic parameters evaluated - subcutaneous cell tissue, PF and intraperitoneal fat, and NAFLD - presented high statistical association with body mass index. NAFLD was observed in eight obese patients (36.36%), and PF and AFI were the variables with highest statistical significance, with p < 0.0001. Ultrasonography is useful tool in the differentiation and quantification of visceral and subcutaneous fat in children. The measures of PF and AFI are useful in the assessment of visceral fat and NAFLD in obese children.


Bovine NK subsets in the afferent lymph and lymph nodes have distinct expression of naïve and activation-associated cell surface expressed molecules, and are differentially stimulated by BCG vaccination.
Bovine natural killer (bNK) cells are heterogeneous cell populations defined by constitutive expression of the natural cytotoxicity receptor, NKp46 (CD335). Two major subsets of bNK cells, classified by differential expression of CD2, display divergent functions in innate immunity, and are hypothesised to contribute to adaptive immunity following vaccination. Here we characterised phenotypic variation of bNK cells within afferent lymph and lymph node (LN) tissues and between CD2+ and CD2- bNK subsets, and report phenotypic changes induced by BCG vaccination. CD2- bNK cells, which dominate in the afferent lymph and LN, displayed lower expression of the activation marker CD25 within the LN, with CD25+ cells being less than half as frequent as in afferent lymph. Furthermore, we found bNK cells had a lower expression of CD45RB, associated in cattle with naïve cell status, within LN compared to afferent lymph. Following BCG vaccination, bNK cells in afferent lymph draining the vaccination site showed increased CD2-CD25+ frequencies and increased expression of CD25 on CD2+ bNK cells, although the frequency of these cells remained unchanged. In summary, we provide an overview of the phenotype of bNK cells within bovine lymphatic tissues, and provide an indication of how subsets may diverge following BCG vaccination.


Primary pulmonary artery sarcoma.
Primary pulmonary artery sarcoma is an uncommon tumor. We report a case of a 73-year-old male patient with a two-week history of palpitations and shortness of breath, aggravated for two days and was believed to be pulmonary hypertension. Emergency heart ultrasound after admission presented a massive pulmonary embolism in the pulmonary artery. The patient's condition was successfully managed with urgent pulmonary artery embolectomy. The patient demonstrated improvement in hemodynamics after the operation. Histologic and immunohistochemical assays were performed and a diagnosis was made as primary pulmonary artery sarcoma arising from the left pulmonary artery. Resection of the tumor is recommended for the treatment of this rare malignant tumor. The corresponding chemotherapy, follow-up and prognosis are described as well in this case report.


[Effectiveness of the fixed combination of type II native collagen, omega-3 and astaxanthin in patients with knee osteoarthritis].
osteoarthritis is one of the most prevalent chronic diseases in the world and is defined as the gradual loss of cartilage in the joints, mainly that of the knee. It is considered a cause of disability in older adults and is characterized by pain, stiffness and loss of mobility. observational study to evaluate the effect of the combination of non-hydrolyzed type II native collagen (CII-NH), omega-3 (Om-3) and astaxanthin (AX), in a population of 182 patients with knee osteoarthritis grade I/II. Measurements of thigh circumference, arcs of movement and pain were obtained through international scales such as the visual analogue pain scale (VAS), the Lequesne index and the Western Ontario McMaster Universities Osteoarthritis Index (WOMAC) scale. Medical check-ups were performed every 30 days for three months. The Statgraphics software (Statgraphics Technologies, Virginia) was used, the evaluation of the variables and the statistical significance were determined by t Student test and the results are shown as a mean. it was shown that daily consumption increases mobility, decreases knee pain and inflammation in patients within three months. Additionally, there was a reduction in the consumption of nonsteroidal anti-inflammatory drugs (NSAIDs) by the study subjects. the fixed combination of non-hydrolyzed type II collagen, omega-3 and astaxanthin, generates, in the short term, a decrease in inflammation and stiffness in patients with osteoarthritis.


Association between response to triptans and response to erenumab: real-life data.
Triptans and erenumab are both migraine-specific agents acting on the calcitonin gene-related peptide pathway. Therefore, response to triptans might be associated with response to erenumab. In our study, consecutive patients referring to the Headache Centers of the Abruzzo region from January 2019 to March 2020 and treated with erenumab were interviewed about past use and efficacy of triptans. Triptan users were classified as 'triptan responders' if they were headache-free 2 h after treating ≥3 migraine attacks with ≥1 triptan. We considered patients as 'erenumab responders', if they had a ≥ 50% mean reduction in monthly migraine days between the 4th and the 6th month from treatment start compared with baseline. Of 91 triptan users, 73 (80.2%) were triptan responders and 58 (63.7%) were erenumab responders. The odds ratio of being erenumab responder was 3.64 (95% CI, 1.25-10.64) for triptan users as compared to non-users. (P = 0.014). Besides, starting erenumab improved triptan response in both erenumab responders and non-responders. Our data of an association between response to triptans and response to erenumab can be useful for patient advice and to improve the understanding of migraine pathophysiology and treatment.


Initiation of multicellular differentiation in Dictyostelium discoideum is regulated by coronin A.
Many biological systems respond to environmental changes by activating intracellular signaling cascades, resulting in an appropriate response. One such system is represented by the social amoeba Dictyostelium discoideum. When food sources become scarce, these unicellular cells can initiate a cAMP-driven multicellular aggregation program to ensure long-term survival. On starvation, the cells secrete conditioned medium factors that initiate cAMP signal transduction by inducing expression of genes such as cAMP receptors and adenylate cyclase. The mechanisms involved in the activation of the first pulses of cAMP release have been unclear. We here show a crucial role for the evolutionarily conserved protein coronin A in the initiation of the cAMP response. On starvation, coronin A-deficient cells failed to up-regulate the expression of cAMP-regulated genes, thereby failing to initiate development, despite a normal prestarvation response. Of importance, external addition of cAMP to coronin A-deficient cells resulted in normal chemotaxis and aggregate formation, thereby restoring the developmental program and suggesting a functional cAMP relay in the absence of coronin A. These results suggest that coronin A is dispensable for cAMP sensing, chemotaxis, and development per se but is part of a signal transduction cascade essential for system initiation leading to multicellular development in Dictyostelium.


Mucosal melanoma: from molecular landscape to current treatment strategies.
Mucosal melanoma (MM) is an aggressive tumor originating from melanocytes located in the respiratory, gastrointestinal, and urogenital tract with clinical and pathologic characteristics distinct from cutaneous melanoma. In addition, MMs have a unique biology that contributes to delayed diagnosis and, therefore an adverse prognosis. The factors all contribute to a treatment paradigm unique from its more studied cutaneous brethren. Due to the rarity of this disease, well-established protocols for the treatment of this pathology have yet to be established. The use of immune checkpoint inhibitors patterned after cutaneous melanoma has become the de facto primary therapeutic approach; however, cytotoxic strategies and pathway-targeted therapies have a defined role in treatment. Judicious use of these approaches can give rise to durable unmaintained disease responses.


An unusual case of multilocular Stafne bone cavity.
Stafne bone defects (SBDs) are asymptomatic lingual bone depressions of the lower jaw that are frequently caused by soft-tissue inclusion. The common variant of SBD exists at the third molar region of the mandible below the inferior dental canal is an and ovoid-shaped homogeneous well-defined radiolucency. In this report, an unusual occurrence of SBD with multilocular appearance is presented. Asymptomatic lingual bone defects may represent various radiographic features. Detailed radiographic evaluation with CT scans should be performed to differentiate SBDs from other pathologies.


Incidence of progressive supranuclear palsy and multiple system atrophy in Olmsted County, Minnesota, 1976 to 1990.
Information on the incidence of progressive supranuclear palsy (PSP) is limited; incidence rates for multiple system atrophy (MSA) are not available. We studied the incidence of PSP and MSA in Olmsted County, Minnesota, for the years 1976 to 1990. This study was part of a larger investigation of all forms of parkinsonism. We used the medical records-linkage system of the Rochester Epidemiology Project to identify all subjects whose records contained documentation of any from of parkinsonism, related neurodegenerative diseases, or tremor of any type. A nurse abstractor screened the records and, when applicable, a neurologist reviewed them to determine the presence or absence of parkinsonism. Cases of parkinsonism were classified using specified diagnostic criteria. Population denominators were derived from census data and were corrected by removing prevalent cases of parkinsonism. Over the 15 years of the study, we found 16 incident cases of PSP and nine incident cases of MSA. No cases of PSP or MSA had onset before age 50 years. The average annual incidence rate (new cases per 100,000 person-years) for ages 50 to 99 years was 5.3 for PSP and 3.0 for MSA. The incidence of PSP increased steeply with age from 1.7 at 50 to 59 years to 14.7 at 80 to 99 years, and was consistently higher in men. Median survival time from symptom onset was 5.3 years for PSP and 8.5 years for MSA. The incidence of PSP increases with age and is consistently higher in men at all ages. PSP and MSA are more common than previously recognized.


Effects of metabolic acidosis on PTH and 1,25(OH)2D3 response to low calcium diet.
To study the effects of chronic metabolic acidosis on the metabolism of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] rats were given either a low calcium diet (LCD) (0.002% calcium) or chow (1.2% calcium); ammonium chloride (NH4Cl) was added (1 or 1.5%) to the drinking water of some rats eating LCD or chow while others served as nonacidotic controls. LCD increased circulating 1,25(OH)2D3 levels from 46 +/- 14 to 204 +/- 24 pg/ml (P less than 0.001) in the absence of NH4Cl; 1.5% NH4Cl prevented the increase in 1,25(OH)2D3 (25 +/- 6 vs. 27 +/- 8 pg/ml (P, NS) but 1% NH4Cl did not (50 +/- 12 vs. 161 +/- 23 pg/ml; P less than 0.001). Acidosis suppressed neither serum immunoreactive parathyroid hormone (PTH) nor urine cAMP response to LCD. Although total serum calcium and phosphorus showed no regular changes with NH4Cl, acidosis raised blood ionized calcium in rats fed either chow or LCD, and serum 1,25(OH)2D3 levels were inversely correlated with ionized calcium (r = 0.714; P less than 0.001) during LCD. Chronic NH4Cl acidosis prevented serum 1,25(OH)2D3 from rising during LCD, independent of changes in PTH secretion, cAMP generation, or serum phosphorus. The absence of a 1,25(OH)2D3 response may be due to increased ionized calcium produced by acidosis.


An expanded regulatory network temporally controls Candida albicans biofilm formation.
Candida albicans biofilms are composed of highly adherent and densely arranged cells with properties distinct from those of free-floating (planktonic) cells. These biofilms are a significant medical problem because they commonly form on implanted medical devices, are drug resistant and are difficult to remove. C. albicans biofilms are not static structures; rather they are dynamic and develop over time. Here we characterize gene expression in biofilms during their development, and by comparing them to multiple planktonic reference states, we identify patterns of gene expression relevant to biofilm formation. In particular, we document time-dependent changes in genes involved in adhesion and metabolism, both of which are at the core of biofilm development. Additionally, we identify three new regulators of biofilm formation, Flo8, Gal4, and Rfx2, which play distinct roles during biofilm development over time. Flo8 is required for biofilm formation at all time points, and Gal4 and Rfx2 are needed for proper biofilm formation at intermediate time points.


The Travelling Particles: Investigating microplastics as possible transport vectors for multidrug resistant E. coli in the Weser estuary (Germany).
The prevalence of multidrug-resistant Gram-negative bacteria in aquatic environments has been a long withstanding health concern, namely extended-spectrum beta-lactamase (ESBL) producing Escherichia coli. Given increasing reports on microplastic (MP) pollution in these environments, it has become crucial to better understand the role of MP particles as transport vectors for such multidrug-resistant bacteria. In this study, an incubation experiment was designed where particles of both synthetic and natural material (HDPE, tyre wear, and wood) were sequentially incubated at multiple sites along a salinity gradient from the Lower Weser estuary (Germany) to the offshore island Helgoland (German Bight, North Sea). Following each incubation period, particle biofilms and water samples were assessed for ESBL-producing E. coli, first by the enrichment and detection of E. coli using Fluorocult® LMX Broth followed by cultivation on CHROMAgar™ ESBL media to select for ESBL-producers. Results showed that general E. coli populations were present on the surfaces of wood particles across all sites but none were found to produce ESBLs. Additionally, neither HDPE nor tyre wear particles were found to harbour any E. coli. Conversely, ESBL-producing E. coli were present in surrounding waters from all sites, 64% of which conferred resistances against up to 3 other antibiotic groups, additional to the beta-lactam resistances intrinsic to ESBL-producers. This study provides a first look into the potential of MP to harbour and transport multidrug-resistant E. coli across different environments and the approach serves as an important precursor to further studies on other potentially harmful MP-colonizing species.


Compensation for variable intrinsic neuronal excitability by circuit-synaptic interactions.
Recent theoretical and experimental work indicates that neurons tune themselves to maintain target levels of excitation by modulating ion channel expression and synaptic strengths. As a result, functionally equivalent circuits can produce similar activity despite disparate underlying network and cellular properties. To experimentally test the extent to which synaptic and intrinsic conductances can produce target activity in the presence of variability in neuronal intrinsic properties, we used the dynamic clamp to create hybrid two-cell circuits built from four types of stomatogastric neurons coupled to the same model Morris-Lecar neuron by reciprocal inhibition. We measured six intrinsic properties (input resistance, minimum membrane potential, firing rate in response to +1 nA of injected current, slope of the frequency-current curve, spike height, and spike voltage threshold) of dorsal gastric, gastric mill, lateral pyloric, and pyloric dilator neurons from male crabs of the species Cancer borealis. The intrinsic properties varied twofold to sevenfold in each cell type. We coupled each biological neuron to the Morris-Lecar model with seven different values of inhibitory synaptic conductance and also used the dynamic clamp to add seven different values of an artificial h-conductance, thus creating 49 different circuits for each biological neuron. Despite the variability in intrinsic excitability, networks formed from each neuron produced similar circuit performance at some values of synaptic and h-conductances. This work experimentally confirms results from previous modeling studies; tuning synaptic and intrinsic conductances can yield similar circuit outputs from neurons with variable intrinsic excitability.


Persistent organic pollutants and early menopause in U.S. women.
Endocrine-disrupting chemicals (EDCs) adversely affect human health. Our objective was to determine the association of EDC exposure with earlier age of menopause. Cross-sectional survey using National Health and Nutrition Examination Survey (NHANES) data from 1999 to 2008 (n = 31,575 females). Eligible participants included: menopausal women >30 years of age; not currently pregnant, breastfeeding, using hormonal contraception; no history of bilateral oophorectomy or hysterectomy. Exposures, defined by serum lipid and urine creatinine-adjusted measures of EDCs, data were analyzed: > 90th percentile of the EDC distribution among all women, log-transformed EDC level, and decile of EDC level. Multi linear regression models considered complex survey design characteristics and adjusted for age, race/ethnicity, smoking, body mass index. EDCs were stratified into long (>1 year), short, and unknown half-lives; principle analyses were performed on those with long half-lives as well as phthalates, known reproductive toxicants. Secondary analysis determined whether the odds of being menopausal increased with EDC exposure among women aged 45-55 years. This analysis examined 111 EDCs and focused on known reproductive toxicants or chemicals with half-lives >1 year. Women with high levels of β-hexachlorocyclohexane, mirex, p,p'-DDE, 1,2,3,4,6,7,8-heptachlorodibenzofuran, mono-(2-ethyl-5-hydroxyhexyl) and mono-(2-ethyl-5-oxohexyl) phthalate, polychlorinated biphenyl congeners -70, -99, -105, -118, -138, -153, -156, -170, and -183 had mean ages of menopause 1.9 to 3.8 years earlier than women with lower levels of these chemicals. EDC-exposed women were up to 6 times more likely to be menopausal than non-exposed women. This study of a representative sample of US women documents an association between EDCs and earlier age at menopause. We identified 15 EDCs that warrant closer evaluation because of their persistence and potential detrimental effects on ovarian function. Earlier menopause can alter the quantity and quality of a woman's life and has profound implications for fertility, human reproduction, and our global society.


Downregulation of Aquaporin3 in Systemic Sclerosis Dermal Fibroblasts.
Skin dryness and thickening are hallmarks of systemic sclerosis (SSc) disease. Aquaporins (AQPs) are plasma membrane proteins that transport glycerol and water, resulting in water retention and skin hydration. Expression of AQPs has been evaluated in human normal skin. However, expression of these proteins in SSc dermal fibroblasts has not yet been reported. The aim of this study was to assess the expression profile of AQPs in dermal fibroblasts of SSc patients. Fibroblast cells were extracted from SSc and healthy skin biopsies and characterized using fibroblast surface protein antibody. The SYBR Green Real-time PCR was used to evaluate the mRNA expression of AQP1, 3, 5, 7, 9, and 10 in dermal fibroblasts. Immunoblotting was performed to confirm the results of Real-time PCR. Our data demonstrated that only AQP1, AQP3, and AQP9 were expressed in human skin fibroblasts. Moreover, the expression of AQP3 mRNA and protein were significantly decreased in SSc dermal fibroblasts compare to healthy fibroblasts. AQP3, which involves in skin hydration and wound healing through water and glycerol transmission, is downregulated in SSc fibroblasts. Based on previous studies and our results, it seems that SSc manifestations like skin dryness, abnormal wound healing, and fibrotic lesions may be related to downregulation of AQP3 in SSc fibroblasts. Therefore, induction of AQP3 expression can be a potential treatment to relieve SSc skin thickness in the future.


Cone Beam Computed Tomography- An Effective Tool in Detecting Caries Under Fixed Dental Prostheses.
Caries under restorations is the most common reason for re-treatment and replacement in restorative failures. To avoid failures of fixed dental prostheses, it is important to diagnose caries under it earlier. Without image degradation and metal artifacts, Cone Beam Computed Tomography (CBCT) can be the solution to detect caries without removing fixed dental prostheses. The aim of this study was to determine the efficacy of CBCT in detecting caries under fixed dental prostheses. Each specimen was scanned with CBCT for evaluation of secondary caries under fixed prostheses. Exposure parameters were 60 kVp and 3mA. Field of View (FOV) used was 8cm X 8cm. According to International Caries Detection and Assessment System (ICDAS) criteria, six Grade 6 carious extracted teeth were selected. All teeth were prepared with four different specimens - full metal, metal-ceramic, full ceramic and metal-acrylic crowns for each tooth. Each specimen was scanned by CBCT. T-test was performed for mean gray value differences between caries and noncaries regions of each material. Gray values were recorded and evaluated for different parameters using two-way analysis of variance. Significant differences were found with respect to material (full metal, metal-ceramic, full ceramic and metal-acrylic) and situation (caries/noncaries) (p<0.001). There were no significant differences with respect to location (anterior or posterior). Mean gray values of caries and noncaries regions were found to be different for each material. CBCT can be used as a post-treatment diagnostic technique for detecting caries under fixed prostheses without removing it.


Effect of aromatherapy on symptoms of dysmenorrhea in college students: A randomized placebo-controlled clinical trial.
The purpose of this study was to explore the effect of aromatherapy on menstrual cramps and symptoms of dysmenorrhea. The study was a randomized placebo-controlled trial. The subjects were 67 female college students who rated their menstrual cramps to be greater than 6 on a 10-point visual analogue scale, who had no systemic or reproductive diseases, and who did not use contraceptive drugs. Subjects were randomized into three groups: (1) an experimental group (n = 25) who received aromatherapy, (2) a placebo group (n = 20), and (3) a control group (n = 22). Aromatherapy was applied topically to the experimental group in the form of an abdominal massage using two drops of lavender (Lavandula officinalis), one drop of clary sage (Salvia sclarea), and one drop of rose (Rosa centifolia) in 5 cc of almond oil. The placebo group received the same treatment but with almond oil only, and the control group received no treatment. The menstrual cramps levels was assessed using a visual analogue scale and severity of dysmenorrhea was measured with a verbal multidimensional scoring system. The menstrual cramps were significantly lowered in the aromatherapy group than in the other two groups at both post-test time points (first and second day of menstruation after treatment). From the multiple regression aromatherapy was found to be associated with the changes in menstrual cramp levels (first day: Beta = -2.48, 95% CI: -3.68 to -1.29, p < 0.001; second day: Beta = -1.97, 95% CI: -3.66 to -0.29, p = 0.02 and the severity of dysmenorrhea (first day: Beta = 0.31, 95% CI: 0.05 to 0.57, p = 0.02; second day: Beta = 0.33, 95% CI: 0.10 to 0.56, p = 0.006) than that found in the other two groups. These findings suggest that aromatherapy using topically applied lavender, clary sage, and rose is effective in decreasing the severity of menstrual cramps. Aromatherapy can be offered as part of the nursing care to women experiencing menstrual cramps or dysmenorrhea.


Targeting Neoantigens for Personalised Immunotherapy.
This review discusses the rapidly evolving field of immunotherapy research, focusing on the types of cancer antigens that can be recognised by the immune system and potential methods by which neoantigens can be exploited clinically to successfully target and clear tumour cells. Recent studies suggest that the likelihood of successful immunotherapeutic targeting of cancer will be reliant on immune response to neoantigens. This type of cancer-specific antigen arises from somatic variants that result in alteration of the expressed protein sequence. Massively parallel sequencing techniques now allow the rapid identification of these genomic mutations, and algorithms can be used to predict those that will be processed by the proteasome, bind to the transporter complex and encode peptides that bind strongly to individual MHC molecules. The emerging data from assessment of the immunogenicity of neoantigens suggests that only a minority of mutations will form targetable epitopes and therefore the potential for immunotherapeutic targeting will be greater in cancers with a higher frequency of protein-altering somatic variants. It is evident that neoantigens contribute to the success of some immunotherapeutic interventions and that there is significant scope for specific targeting of these antigens to develop new treatment approaches.


Isocoumarin formation by heterologous gene expression and modification by host enzymes.
Heterologous expression has been proven to be a successful strategy for the identification of metabolites encoded by cryptic/silent genes. Expression of a nonreducing polyketide synthase (NR-PKS) gene from Penicillium crustosum in Aspergillus nidulans led to the accumulation of three isocoumarins 1-3. Feeding experiments revealed that the PKS product 1 can be converted by the host enzymes to its hydroxylated (2) and methylated (3) derivatives. These results provided one additional example that unexpected further modifications of an enzyme product can take place in a heterologous host.


Differential responses of benthic microbes and meiofauna to fish-farm disturbance in coastal sediments.
Bacterial and meiofaunal abundance and biomass and their response to the disturbance induced by fish-farm biodeposition were investigated from March to October 1997 on a monthly basis at two stations of the Gaeta Gulf (Tyrrhenian Sea, Mediterranean Sea). The biopolymeric fraction of the organic matter was characterized by high concentrations which was similar at both fish-farming-impacted and control stations. Similarly, bacteria accounted for a small fraction of the biopolymeric organic carbon (< 1%), while the contribution due to auto-fluorescent cell biomass (i.e. prokaryotic and eukaryotic cells displaying auto-fluorescence) to the total biopolymeric carbon was quantitatively negligible (< 0.1%). Benthic bacteria appear to be sensitive to organic enrichment as their abundance increased significantly beneath the cage, whilst numbers of meiofauna was lower than in the control. Changes occurred also in terms of individual nematode biomass that increased as result of the biodeposition. A particularly useful tool appeared to be represented by the ratio of benthic auto-fluorescent cells to bacterial abundance, bacteria to meiofaunal biomass and auto-fluorescent cells to meiofauna biomass. All these parameters described well the impact due to biodeposition on the benthic environment as their ratios displayed significantly higher values in farm sediments, but recovered rapidly (15 days) to values observed in the control (i.e. undisturbed conditions) immediately after cage removal. Changes observed in the present study highlight that the increased organic loading determined a shift of the relative contribution of the different benthic components to the total biopolymeric carbon, so that in highly impacted systems total benthic biomass becomes increasingly dominated by microbial components.


Toward understanding the anticorrosive mechanism of some thiourea derivatives for carbon steel corrosion: A combined DFT and molecular dynamics investigation.
The mutually corroborated density functional theory (DFT) and molecular dynamics (MD) simulation methodology were employed to evaluate the inhibition performance of three thiourea derivatives (Inh1, Inh2, and Inh3) on carbon steel corrosion. Experimental results have shown that the corrosion rate follows the order: Inh3>Inh2>Inh1. Quantum chemical descriptors such as the frontier orbital energies (EHOMO and ELUMO), the energy gap between ELUMO and EHOMO (ΔE), dipole moment (μ), and Fukui index have been calculated and discussed. Some significant factors such as solvent, temperature, and coverage have been considered when investigating the adsorption of aforementioned thiourea derivatives on Fe(110) surface. Our results provide important atomic/molecular insights into the anticorrosive mechanism of inhibitor molecules, which could help in understanding the organic-metal interface and designing more appropriate organic corrosion inhibitors.


Spectral interferometry with waveform-dependent relativistic high-order harmonics from plasma surfaces.
The interaction of ultra-intense laser pulses with matter opened the way to generate the shortest light pulses available nowadays in the attosecond regime. Ionized solid surfaces, also called plasma mirrors, are promising tools to enhance the potential of attosecond sources in terms of photon energy, photon number and duration especially at relativistic laser intensities. Although the production of isolated attosecond pulses and the understanding of the underlying interactions represent a fundamental step towards the realization of such sources, these are challenging and have not yet been demonstrated. Here, we present laser-waveform-dependent high-order harmonic radiation in the extreme ultraviolet spectral range supporting well-isolated attosecond pulses, and utilize spectral interferometry to understand its relativistic generation mechanism. This unique interpretation of the measured spectra provides access to unrevealed temporal and spatial properties such as spectral phase difference between attosecond pulses and field-driven plasma surface motion during the process.


An enzyme-inorganic hybrid nanoflower based immobilized enzyme reactor with enhanced enzymatic activity.
A facile approach for the synthesis of enzyme-inorganic hybrid nanoflowers and their application as an immobilized α-chymotrypsin (ChT) reactor (IMER) for highly efficient protein digestion was described. The hybrid nanoflowers were room-temperature synthesized in aqueous solution using calcium phosphate (Ca3(PO4)2) as the inorganic component and ChT as the organic component. The effects of reaction parameters on the formation of the enzyme-embedded hybrid nanoflowers and their growth mechanism were investigated systematically. By monitoring the reaction of N-benzoyl-l-tyrosine ethyl ester (BTEE), the enzymatic activity of the immobilized ChT was calculated and the results showed 266% enhancement in enzymatic activity. The performance of such a nanoreactor was further demonstrated by digesting bovine serum albumin (BSA) and human serum albumin (HSA), with a stringent threshold for unambiguous identification of these digests, the yielding sequence coverages for nanoflower-based digestion were 48% and 34%, higher than those obtained with the free enzyme. The digestion time of BSA and HSA in the former case was less than 2 min, about 1/360 of that performed in the latter case (12 h). Furthermore, the residual activity of the nanoflowers decreased slightly even after eight repeated use, demonstrating promising stability. In addition, the hybrid nanoflower-based IMER was applicable to the digestion of a complex human sample, showing great promise for proteome analysis.


Circulating syndecan-1 is reduced in pregnancies with poor fetal growth and its secretion regulated by matrix metalloproteinases and the mitochondria.
Fetal growth restriction is a leading cause of stillbirth that often remains undetected during pregnancy. Identifying novel biomarkers may improve detection of pregnancies at risk. This study aimed to assess syndecan-1 as a biomarker for small for gestational age (SGA) or fetal growth restricted (FGR) pregnancies and determine its molecular regulation. Circulating maternal syndecan-1 was measured in several cohorts; a large prospective cohort collected around 36 weeks' gestation (n = 1206), a case control study from the Manchester Antenatal Vascular service (285 women sampled at 24-34 weeks' gestation); two prospective cohorts collected on the day of delivery (36 + 3-41 + 3 weeks' gestation, n = 562 and n = 405 respectively) and a cohort who delivered for preterm FGR (< 34 weeks). Circulating syndecan-1 was consistently reduced in women destined to deliver growth restricted infants and those delivering for preterm disease. Syndecan-1 secretion was reduced by hypoxia, and its loss impaired proliferation. Matrix metalloproteinases and mitochondrial electron transport chain inhibitors significantly reduced syndecan-1 secretion, an effect that was rescued by coadministration of succinate, a mitochondrial electron transport chain activator. In conclusion, circulating syndecan-1 is reduced among cases of term and preterm growth restriction and has potential for inclusion in multi-marker algorithms to improve detection of poorly grown fetuses.


Characterization of a putative ancestor of coxsackievirus B5.
Like other RNA viruses, coxsackievirus B5 (CVB5) exists as circulating heterogeneous populations of genetic variants. In this study, we present the reconstruction and characterization of a probable ancestral virion of CVB5. Phylogenetic analyses based on capsid protein-encoding regions (the VP1 gene of 41 clinical isolates and the entire P1 region of eight clinical isolates) of CVB5 revealed two major cocirculating lineages. Ancestral capsid sequences were inferred from sequences of these contemporary CVB5 isolates by using maximum likelihood methods. By using Bayesian phylodynamic analysis, the inferred VP1 ancestral sequence dated back to 1854 (1807 to 1898). In order to study the properties of the putative ancestral capsid, the entire ancestral P1 sequence was synthesized de novo and inserted into the replicative backbone of an infectious CVB5 cDNA clone. Characterization of the recombinant virus in cell culture showed that fully functional infectious virus particles were assembled and that these viruses displayed properties similar to those of modern isolates in terms of receptor preferences, plaque phenotypes, growth characteristics, and cell tropism. This is the first report describing the resurrection and characterization of a picornavirus with a putative ancestral capsid. Our approach, including a phylogenetics-based reconstruction of viral predecessors, could serve as a starting point for experimental studies of viral evolution and might also provide an alternative strategy for the development of vaccines.


[Role of IL-22 in the pathogenesis of skin diseases].
IL-22 is an IL-10 family cytokine that acts mainly on epithelial cells. It is produced by immune cell subsets, including CD4⁺ T cells, natural killer cells, and natural killer T cells. In the skin, IL-22 mediates keratinocyte proliferation and epidermal hyperplasia, inhibits terminal differentiation of keratinocytes, and induces the production of antimicrobial proteins. Although IL-22 production was initially linked with IL-17 expression in Th17 cells, IL-22 production can also occur in an apparently unique subset of cells that lacks the production of IL-17 and IFN-γ (Th22). Interestingly, Th22 cells express skin homing chemokine receptors CCR4 and CCR10. Indeed, Th22 cells reside in the normal skin and are shown to be enriched in the lesional skin of inflammatory skin diseases, indicating the importance of IL-22 in skin homeostasis and pathogenesis of skin diseases. Although psoriasis is the first example of an organ-specific immune disorder for which the role of IL-22 has been comprehensively studied, a growing body of evidence indicates that this cytokine also plays a critical role in the pathogenesis of atopic dermatitis. In this review, we discuss the role of IL-22 in the pathogenesis of skin diseases, particularly focusing on psoriasis and atopic dermatitis. Targeting IL-22 may have promise as a potential therapeutic for various skin diseases.


Reducing early smokers' risk for future smoking and other problem behavior: insights from a five-year longitudinal study.
To identify risk and protective factors during early and later adolescence that predict future regular smoking and multiple problem behavior among at-risk youth, defined as those who tried smoking by grade 7. At grades 7, 10, and 12, data were collected from 2,000 early smokers drawn from California and Oregon. Multivariate regression analyses tested predictors of the two grade 12 outcomes in separate models using data from grades 7 and 10. Gender interactions and buffering of risk factors by protective factors were assessed. For at-risk youth, consistent protective factors against future smoking and problem behavior included living in an intact nuclear family (all four models) plus getting good grades and parental disapproval of smoking/drug use (three of four models). Consistent risk factors included exposure to substance-using peers (four models) and problems in school (three of four models). Adult substance use was a predictor during early, but not later, adolescence; pro-smoking/drug use beliefs were significant predictors during later adolescence. There were few differences across gender and no significant buffers against risk. At-risk youth would likely benefit from peer resistance training, parental involvement in prevention efforts, and efforts to improve educational performance during both middle school and high school. Changing pro-drug beliefs may be more effective among older adolescents.


Population fraction of Parkinson's disease attributable to preventable risk factors.
Parkinson's disease is the fastest growing neurologic disease with seemingly no means for prevention. Intrinsic risk factors (age, sex, genetics) are inescapable, but environmental factors are not. We studied population attributable fraction and estimated fraction of PD that could be reduced if modifiable risk factors were eliminated. Assessing several known risk factors simultaneously in one study, we demonstrate that all were operative and independent, underscoring etiological heterogeneity within a single population. We investigated repeated blows to head in sports or combat as a potential new risk factor, and found it was associated with two-fold increased risk of PD. Considering modifiable risk factors, 23% of PD cases in females were attributable to pesticides/herbicides exposure, and 30% of PD cases in males was attributable to pesticides/herbicides, Agent Orange/chemical warfare, and repeated blows to the head. Thus, one-in-three cases of PD in males, and one-in-four cases in females could have potentially been prevented.


Genome-Wide Analysis of the DC1 Domain Protein Gene Family in Tomatoes under Abiotic Stress.
DC1 (Divergent C1) domain proteins are a new class of proteins that have been discovered in recent years, which play an important role in plant growth, development, and stress response. In order to better study the distribution and function of DC1 domain proteins in tomatoes, a genome-wide identification was conducted. It was found that there are twenty-one DC1 domain protein genes distributed on nine chromosomes of tomatoes, named SlCHP1-21. Phylogenetic analysis shows that twenty-one SlCHP genes are divided into six subfamilies. Most of the SlCHP genes in tomatoes have no or very short introns. All SlCHP proteins, with the exception of SlCHP8 and SlCHP17, contain variable amounts of C1 domain. Analysis of the SlCHP gene promoter sequence revealed multiple cis-elements responsive to plant stress. qRT-CR analysis showed that most members of SlCHP gene expressed in the roots. The SlCHP11, 13, 16, 17, and SlCHP20 genes showed specific responses to high temperature, low temperature, salt, and drought stress. In addition, the subcellular localization and interaction proteins of SlCHP were analyzed and predicted. Together, these results provides a theoretical basis for further exploration of the function and mechanism of the SlCHP gene in tomatoes.


Invertebrate models of dystonia.
The neurological movement disorder dystonia is an umbrella term for a heterogeneous group of related conditions where at least 20 monogenic forms have been identified. Despite the substantial advances resulting from the identification of these loci, the function of many DYT gene products remains unclear. Comparative genomics using simple animal models to examine the evolutionarily conserved functional relationships with monogenic dystonias represents a rapid route toward a comprehensive understanding of these movement disorders. Current studies using the invertebrate animal models Caenorhabditis elegans and Drosophila melanogaster are uncovering cellular functions and mechanisms associated with mutant forms of the well-conserved gene products corresponding to DYT1, DYT5a, DYT5b, and DYT12 dystonias. Here we review recent findings from the invertebrate literature pertaining to molecular mechanisms of these gene products, torsinA, GTP cyclohydrolase I, tyrosine hydroxylase, and the alpha subunit of Na+/K ATPase, respectively. In each study, the application of powerful genetic tools developed over decades of intensive work with both of these invertebrate systems has led to mechanistic insights into these human disorders. These models are particularly amenable to large-scale genetic screens for modifiers or additional alleles, which are bolstering our understanding of the molecular functions associated with these gene products. Moreover, the use of invertebrate models for the evaluation of DYT genetic loci and their genetic interaction networks has predictive value and can provide a path forward for therapeutic intervention.


Triptolide-induced transcriptional arrest is associated with changes in nuclear substructure.
Triptolide, an active component of the medicinal herb lei gong teng, is a potent anticancer and anti-inflammatory therapeutic. It potently inhibits nuclear factor-kappaB transcriptional activation after DNA binding, although a precise mechanism is as yet unknown. Here, we report that triptolide also induces distinct nuclear substructural changes in HeLa cells. These changes in the nucleolus and nuclear speckles are reversible and dependent on both time and concentration. Furthermore, nuclear changes occurred within hours of triptolide treatment and were calcium and caspase independent. Rounding of nuclear speckles, an indication of transcriptional arrest, was evident and was associated with a decrease in RNA polymerase II (RNA Pol II) COOH-terminal domain Ser(2) phosphorylation. Additionally, the nucleolus disassembled and RNA Pol I activity declined after RNA Pol II inhibition. We therefore conclude that triptolide causes global transcriptional arrest as evidenced by inactivity of RNA Pol I and II and the subsequent alteration in nuclear substructure.


Non-surgical treatment of hip osteoarthritis. Hip school, with or without the addition of manual therapy, in comparison to a minimal control intervention: protocol for a three-armed randomized clinical trial.
Hip osteoarthritis is a common and chronic condition resulting in pain, functional disability and reduced quality of life. In the early stages of the disease, a combination of non-pharmacological and pharmacological treatment is recommended. There is evidence from several trials that exercise therapy is effective. In addition, single trials suggest that patient education in the form of a hip school is a promising intervention and that manual therapy is superior to exercise. This is a randomized clinical trial. Patients with clinical and radiological hip osteoarthritis, 40-80 years of age, and without indication for hip surgery were randomized into 3 groups. The active intervention groups A and B received six weeks of hip school, taught by a physiotherapist, for a total of 5 sessions. In addition, group B received manual therapy consisting of joint manipulation and soft-tissue therapy twice a week for six weeks. Group C received a self-care information leaflet containing advice on "live as usual" and stretching exercises from the hip school. The primary time point for assessing relative effectiveness is at the end of the six weeks intervention period with follow-ups after three and 12 months.Primary outcome measure is pain measured on an eleven-point numeric rating scale. Secondary outcome measures are the hip dysfunction and osteoarthritis outcome score, patient's global perceived effect, patient specific functional scale, general quality of life and hip range of motion. To our knowledge this is the first randomized clinical trial comparing a patient education program with or without the addition of manual therapy to a minimal intervention for patients with hip osteoarthritis. ClinicalTrials NCT01039337.


Renal ultrasonographic abnormalities are associated with low glomerular filtration rate calculated by scintigraphy in dogs.
Ultrasound provides information on kidney morphology, but studies relating structural and functional abnormalities in chronic kidney disease (CKD) are lacking. The aim of this descriptive cross-sectional study was to compare individual kidney (IK) B-mode ultrasound abnormalities to IK glomerular filtration rate (GFR) estimated by scintigraphy normalized to plasma volume (PV) in dogs, to evaluate if ultrasonographic findings were associated with low IKGFR/PV. Eighty privately owned dogs with and without clinical suspicion of CKD were prospectively enrolled, and kidney ultrasound and IKGFR/PV were evaluated independently. Ultrasound images were assessed retrospectively for subjective abnormalities (shape, cortical, and medullary hyperechogenicity), and kidney size was measured. The normal IKGFR/PV cutoff was derived from dogs in the study group with no history and clinical signs of kidney disease and normal blood and urine results (n = 28) and was 16.84 mL/min/L. Kidneys were categorized into normal, mild, moderate, and severe ultrasound changes according to subjective ultrasound grades. Associations were found between low IKGFR/PV and abnormal kidney shape (P = .0004), cortical hyperechogenicity (P = .0008), medullary hyperechogenicity (P < .0001), and low kidney volume (P = .0092). Apart from the moderate and severe category comparison, IKGFR/PV value significantly decreased with increasing severity of category. The combination of ultrasonographic subjective abnormalities had a high sensitivity (93.8%) and moderate specificity (65.7%) for detecting low IKGFR/PV. Kidneys with normal IKGFR/PV had a low frequency of mild ultrasound changes. Findings indicate kidneys with increasing number and grade of subjective ultrasound abnormalities are more likely to have a lower IKGFR/PV.


Tissue-specific pretranslational regulation of complement production in human mononuclear phagocytes.
The net production of the complement protein C2, C4, and factor B differ among mononuclear phagocytes from peripheral blood and different tissues in experimental animals and in humans. To examine the mechanisms that regulate these differences in humans, the proportions of C2-producing cells, the average single-cell production rate of C2, the posttranslational glycosylation and kinetics of secretion of C2 and factor B, and the amounts of C2 and factor B mRNA were examined in freshly isolated peripheral blood monocytes, monocytes maintained up to 2 wk in culture, and freshly isolated tissue macrophages from breast milk and bronchoalveolar lavage. In addition, the biosynthesis of two other proteins synthesized and secreted by mononuclear phagocytes, C3 and lysozyme, were examined. We report that despite comparable rates of C3 and lysozyme synthesis and similar processing and kinetics of secretion of C2 and factor B, the freshly isolated tissue macrophage differs from the monocyte-derived macrophage in the proportion of C2-producing cells, in the average single-cell production rate of complement, and in the amounts of specific C2 and factor B mRNA. These differences are tissue specific, because C2-specific mRNA content in bronchoalveolar macrophages is considerably greater than in breast milk macrophages, although the amounts of factor B mRNA are comparable. These data suggest that tissue-specific regulation of complement production in human mononuclear phagocytes occurs at a pretranslational level. These studies now provide a basis for investigation of the molecular effects of agents that modulate the biologic functions of monocytes and macrophages.


Localization of factor-VIII-related antigen in human vascular subendothelium.
Factor-VIII-related antigen has previously been shown to be synthesized by vascular endothelial cells. Using both an immunofluorescent staining technique and electron microscopy, we have demonstrated the presence of factor-VIII-related antigen in human vascular subendothelium. This finding may have implications in the mechanism of platelet adhesion to deendothelialized blood vessel surfaces.


A comparison of three programming languages for a full-fledged next-generation sequencing tool.
elPrep is an established multi-threaded framework for preparing SAM and BAM files in sequencing pipelines. To achieve good performance, its software architecture makes only a single pass through a SAM/BAM file for multiple preparation steps, and keeps sequencing data as much as possible in main memory. Similar to other SAM/BAM tools, management of heap memory is a complex task in elPrep, and it became a serious productivity bottleneck in its original implementation language during recent further development of elPrep. We therefore investigated three alternative programming languages: Go and Java using a concurrent, parallel garbage collector on the one hand, and C++17 using reference counting on the other hand for handling large amounts of heap objects. We reimplemented elPrep in all three languages and benchmarked their runtime performance and memory use. The Go implementation performs best, yielding the best balance between runtime performance and memory use. While the Java benchmarks report a somewhat faster runtime than the Go benchmarks, the memory use of the Java runs is significantly higher. The C++17 benchmarks run significantly slower than both Go and Java, while using somewhat more memory than the Go runs. Our analysis shows that concurrent, parallel garbage collection is better at managing a large heap of objects than reference counting in our case. Based on our benchmark results, we selected Go as our new implementation language for elPrep, and recommend considering Go as a good candidate for developing other bioinformatics tools for processing SAM/BAM data as well.


A Concept Analysis of Synergy for Nursing Education.
Background and Purpose: Synergy is a term found widely throughout the sciences. There is a need for improved methods and delivery of instruction that supports learners in their pursuit of academic goals and facilitates their transition into the professional role. Critical for students is teamwork training, both as team members and facilitators. Some teams have outcomes greater than their own individual efforts, while others do not. One possible explanation for this phenomenon is synergy. A concept analysis was conducted to define synergy in the context of nursing education and to gain insight on its impact on learning. Using Walker and Avant's (2011) method, definitions, uses, attributes, antecedents, consequences, and empirical referents were identified. Methods: Six databases were searched, using the keyword synergy (syntactic variations) and search terms (nursing, education, student, learning, training, collaboration, teamwork, and cooperation) with the parameter of English language (1980-2017). Articles were delimited through duplication removal, title, and abstract review. Thirty articles were selected for review based upon the use of synergy within the context of education. Results: Synergy is demonstrated whengroup outcomes exceed the results of individual efforts. Attributes are mutuality of support, actions, goalswith greater than individual effects, and teaching strategies. Consequences are greater problem-solving, creativity, and goal attainment than are accomplished individually. Implications for Practice: Incorporating synergy into nursing education can promote teamwork and collaboration and facilitate learners to collaborate inter-professionally. Recommendations are to research synergy within nursing education.


Spectrum of metastatic and nonmetastatic skeletal findings with dual-phase 18F-FECH PET/CT in patients with biochemical relapse of prostate cancer.
The aim of this study was to evaluate the spectrum of skeletal findings on dual-phase fluorine-18-fluoroethylcholine (F-FECH) PET/CT performed during the work-up of patients referred for suspected prostate cancer relapse. Three hundred F-FECH PET/CT scans were evaluated prospectively. The low-dose CT features of all cases were categorized as isodense, sclerotic, lytic or mixed lytic/sclerotic and maximum standardized uptake value (SUVmax) values were calculated. Findings on F-FECH PET/CT were correlated with Technetium-99m-methylene diphosphonate planar bone scans and serum prostate-specific antigen. Patient age range was 50-90 years (median 71 years) and prostate-specific antigen values were in the range 0.04-372 ng/ml (Roche Modular method). Seventy-two lesions were detected on F-FECH PET/CT in 45 patients, including 31 (43%) in the pelvis, 17 (23%) in the spine (cervical 3, thoracic 8 and lumbar spine 6) and 10 (13%) in the ribs. Evaluation of low-dose CT in combination with PET helped to characterize benign findings in 21 (29%) lesions. The SUVmax for all except one benign lesion ranged from 0.49 to 2.15. In 51 (71%) lesions because of metastatic disease, SUVmax was 0.6-11.6 for those classified as sclerotic on low-dose CT, 0.7-8.58 for lytic lesions, 1.1-7.65 for isodense lesions and 1.27-3.53 for mixed lytic/sclerotic lesions. Of the 56 F-FECH-avid lesions, 21 lesions showed avidity on bone scan [3 (23%) of the 13 isodense lesions, 14 (40%) of the 35 sclerotic lesions, 2 (50%) of the lytic lesions and 2 (50%) of the mixed sclerotic/lytic lesions]. F-FECH PET/CT identified bone lesions in 15% of patients with suspected prostate cancer relapse. SUVmax in isolation cannot be used to characterize these lesions as benign or malignant. Minimal overlap of benign and malignant lesions was observed above SUVmax of 2.5. Low-dose CT of PET/CT is a useful tool to aid characterization.


Scaling-up anti-predator phenotypic responses of prey: impacts over multiple generations in a complex aquatic community.
Non-consumptive effects (NCEs) of predators owing to induced changes in prey traits are predicted to influence the structure of ecological communities. However, evidence of the importance of NCEs is limited primarily to simple systems (e.g. two to four species) over relatively short periods (e.g. less than one generation). We examined the NCEs of a fish predator, arising from phenotypic plasticity in zooplankton prey traits, over multiple generations of a diverse zooplankton community. The presence of fish, caged to remove consumptive effects, strongly influenced zooplankton community structure, through both direct and indirect NCE pathways, altering the abundance of many taxa by magnitudes as large as 3 to 10-fold. Presence of fish affected different species of cladocerans and copepods both positively and negatively. A particularly striking result was the reversal of dominance in copepod taxa: presence of fish reduced the ratio of calanoids to cyclopoids from 6.3 to 0.43. Further, the NCE of fish had a strong negative trophic cascade to zooplankton resources (phytoplankton). To our knowledge, this is the first experiment to show that NCEs can influence the abundance of multiple prey species over time spans of multiple prey generations. Our findings demonstrate that adaptive phenotypic plasticity of individuals can scale-up to affect the structure of ecological communities.


lncRNA CCAT1 promotes bladder cancer cell proliferation, migration and invasion.
To study the expression patterns of long noncoding RNA (lncRNA) colon cancer-associated transcript 1 (CCAT1) and the changes in cell proliferation, apoptosis, migration and invasion induced by silencing CCAT1 in bladder cancer cells. The expression levels of CCAT1 were determined using realtime quantitative polymerase chain reaction in cancerous tissues and paired normal tissues from 34 patients with bladder cancer. The relationship between clinical characteristics and CCAT1 expression was analyzed. And then we conducted cell experiments. Bladder urothelial carcinoma cell lines T24 and 5637 cells were transfected with CCAT1 small interfering RNA (siRNA) or scramble siRNA. Cell proliferation and apoptosis changes were determined using a Cell Counting Kit-8 (CCK-8) assay and a fl ow cytometry assay. Migration and invasion changes were measured using a wound healing assay and a trans-well assay. microRNAs (miRNAs) were predicted by Starbase 2.0, and their differential expression levels were studied. CCAT1 was signifi cantly upregulated in bladder cancer (P < 0.05). CCAT1 upregulation was positively related to tumor stage (P = 0.004), tumor grade (P = 0.001) and tumor size (P = 0.042). Cell proliferation, migration and invasion were promoted by abnormally expressed CCAT1. miRNAs miR-181b-5p, miR-152-3p, miR-24-3p, miR-148a-3p and miR-490-3p were potentially related to the aforementioned functions of CCAT1. CCAT1 plays an oncogenic role in urothelial carcinoma of the bladder. In addition, CCAT1 may be a potential therapeutic target in this cancer.


[Chronic pelvic pain in women].
This paper reviews empirical and clinical evidence of the aetiology and treatment of medically unexplained chronic pelvic pain in women. Clinical experience from an ongoing randomised treatment trial supplemented by computer-assisted reviews of studies obtained by a Premedline and Medline search (1996 to February 2002) and data from the Cochrane Database of Systematic Reviews and the EBM database of Abstracts of Reviews of Effectiveness. The aetiology of medically unexplained chronic pelvic pain is disputed but likely to be multifactorial. A history of interpersonal difficulties and a stressful life is common, and comorbid psychiatric disorders occur frequently. No treatment of choice emerges from the few controlled treatment trials, though a flexible biopsychosocial approach seems the most promising. Empathic medical evaluation and follow-up within a biopsychosocial framework is recommended. Analgesic, hormonal and, if appropriate, surgical treatment can relieve pain. Additional benefits may be obtained by adding sensory awareness-directed physiotherapy aimed at changing painful muscle tensions, body attitude, movement pattern and dysfunctional respiration pattern. Co-morbid psychiatric disorders should be diagnosed and treated. Cognitive-behavioural stress management intervention aimed at improving coping with pain and current life-situation may be indicated in a subsample of patients.


Symmetrical multifocal liposarcoma.
Multiple liposarcomas are extremely rare. We report on a patient with liposarcoma of the right lower limb who had undergone surgery and radiotherapy 4 years previously for a liposarcoma in the left lower limb. An outline of the management is discussed and a short review of available literature is given.


Treatment of patients with advanced cancer of the prostate: phase III trial, zoladex against castration; a study of the British Prostate Group.
The interim results of the randomised, Phase III trial of Zoladex against castration in the management of patients with metastatic carcinoma of the prostate is discussed. Trials commenced in October 1984 and incorporated 359 patients when recruitment ceased in January 1986. The preliminary report concerns the first 240 patients who had a minimum of 3 months follow-up. Entry criteria included patients who had no previous treatment with the exception of first-line localised radiotherapy and those who had distant bone or soft tissue metastases. Fourteen patients were excluded on the basis of protocol violations. The objective assessment of response was based on the British Prostate Group Criteria and was performed monthly for the first 3 months and 3-monthly thereafter. Pre-treatment disease characteristics of patients in both groups were similar at entry and there were no significant differences in the subjective response data of patients between the orchidectomy (n = 106) and the Zoladex group (n = 120). Objective response rates at 12 and 24 weeks of treatment were also identical for both treatment groups. Serum testosterone concentrations were below the 'castrate' level (less than 2 nmol/L) for Zoladex group as well as the orchidectomy group up to 48 weeks. The drug was well-tolerated with minimal side effects, those resulting from testosterone withdrawal were similar in both groups. The report therefore indicates clearly that this partical formulation of LH-RH analogue provides a valuable alternative to the surgical procedure in the treatment of carcinoma of the prostate.


[Teleradiotherapy of laryngeal cancer using a gas hypoxic mixture (GHM-10)].
A total of 96 laryngeal cancer patients were studied. A comparative analysis of short-term therapeutic results was performed in 2 similar groups: patients receiving radiotherapy without GHM-10 (50) and those receiving radiotherapy with the use of GHM-10 containing 10% of oxygen and 90% of nitrogen (46). Irradiation methods and general tactics of the patients management in both groups were identical. No differences in the short-term results were noted. Radiation reactions of the healthy tissues were less pronounced and noted less frequently in the patients on radiotherapy with the use of GHM-10.


Evaluation of event-free survival as a robust end point in untreated acute myeloid leukemia (Alliance A151614).
Event-free survival (EFS) is controversial as an end point for speeding approvals in newly diagnosed acute myeloid leukemia (AML). We aimed to examine the robustness of EFS, specifically timing of complete remission (CR) in defining induction failure and impact of hematopoietic cell transplantation (HCT). The study included 1884 untreated AML patients enrolled across 5 trials conducted through Alliance for Clinical Trials in Oncology using anthracycline and cytarabine induction chemotherapy. EFS was defined as time from randomization/registration to induction failure, relapse, or death. Three definitions of induction failure were evaluated: failure to achieve CR by 60 days after randomization/registration, failure to achieve CR by the end of all protocol-defined induction courses, and failure to achieve CR by the end of all protocol-defined treatment. We considered either censoring or no censoring at time of non-protocol-mandated HCT. Although relapse and death are firm end points, the determination of induction failure was not consistent across studies. There was minimal impact of censoring at HCT on EFS estimates; however, median EFS estimates differed considerably based on the timing of CR in defining induction failure, with the magnitude of difference being large enough in most cases to lead to incorrect conclusions about efficacy in a single-arm trial, if the trial definition was not consistent with the definition used for the historical control. Timing of CR should be carefully examined in the historical control data used to guide the design of single-arm trials using EFS as the primary end point. Trials were registered at www.clinicaltrials.gov as #NCT00085124, #NCT00416598, # NCT00651261, #NCT01238211, and #NCT01253070.


A network analysis of global cephalopod trade.
The global trade in cephalopods is a multi-billion dollar business involving the fishing and production of more than ten commercially valuable species. It also contributes, in whole or in part, to the subsistence and economic livelihoods of thousands of coastal communities around the world. The importance of cephalopods as a major cultural, social, economic, and ecological resource has been widely recognised, but research efforts to describe the extent and scope of the global cephalopod trade are limited. So far, there are no specific regulatory and monitoring systems in place to analyse the traceability of the global trade in cephalopods at the international level. To understand who are the main global players in cephalopod seafood markets, this paper provides, for the first time, a global overview of the legal trade in cephalopods. Twenty years of records compiled in the UN COMTRADE database were analysed. The database contained 115,108 records for squid and cuttlefish and 71,659 records for octopus, including commodity flows between traders (territories or countries) weighted by monetary value (USD) and volume (kg). A theoretical network analysis was used to identify the emergent properties of this large trade network by analysing centrality measures that revealed key insights into the role of traders. The results illustrate that three countries (China, Spain, and Japan) led the majority of global market movements between 2000 and 2019. Based on volume and value, as well as the number of transactions, 11 groups of traders were identified. The leading cluster consisted of only eight traders, who dominated the cephalopod market in Asia (China, India, South Korea, Thailand, and Vietnam), Europe (the Netherlands, and Spain), and the USA. This paper identifies the countries and territories that acted as major importers or exporters, the best-connected traders, the hubs or accumulators, the modulators, the main flow routes, and the weak points of the global cephalopod trade network over the last 20 years. This knowledge of the network is crucial to move towards an environmentally sustainable, transparent, and food-secure global cephalopod trade.


Pathological and toxicological findings in glyphosate-surfactant herbicide fatality: a case report.
Glyphosate herbicide is promoted by the manufacturer as having no risks to human health, with acute toxicity being very low in normal use. In Thailand, however, poisoning from glyphosate agricultural herbicides has been increasing. A case of rapid lethal intoxication from glyphosate-surfactant herbicide involved a 37-year-old woman, who deliberately ingested approximately 500 mL of concentrated Roundup formulation (41% glyphosate as the isopropylamine salt and 15% polyoxyethylene amine; Mosanto Company). The postmortem examination revealed that the stomach contained 550 mL of yellow fluid. The gastric mucosa of anterior fundus revealed hemorrhage and the small intestines had marked dilatation and thin walls. We used the high-performance liquid chromatography method for determination of serum and gastric content levels of glyphosate. The glyphosate levels of serum and gastric content were 3.05 and 59.72 mg/mL, respectively. Toxic effects of polyoxyethylene amine and Roundup were caused by their ability to erode tissues including mucous membranes and linings of the gastrointestinal and respiratory tracts. A mild degree of pulmonary congestion and edema was observed in both lungs. We proposed that the characteristic picture of microvesicular steatosis of the hepatocytes, seen predominantly in centrilobular zones of the liver, resembled drug-induced hepatic toxicity or secondary hypoxic stress.


The Bleph and the Brain: The Effect of Upper Eyelid Surgery on Chronic Headaches.
To determine effect of upper eyelid surgery on headache symptoms. Consecutive adults undergoing upper eyelid surgery for obscuration of superior visual field, who also reported headache symptoms for greater than 1 year completed a pre- and postoperative Headache Impact Test-6 quality of life questionnaire (study group). A cohort of patients undergoing other oculoplastic procedures with headaches also completed the questionnaire pre- and postoperatively (control group). The study was conducted over a 2-year period. Neither the patients nor the study investigators were masked. Twenty-eight patients met criteria for the study group, and 19 patients in the control group. Mean age was 58.7 and 60.7 years, respectively. There was no statistically significant difference in preoperative location of headaches. There was no significant difference in mean Headache Impact Test-6 scores preoperatively, 57.7 study group, 58.1 control group, p = 0.86. Mean postoperative scores were lower (improved) in the study arm, 45.3, as compared with the control arm, 58.6, p < 0.05. There was no statistically significant difference between individual preoperative survey questions between the study arm and control group, while every Headache Impact Test-6 question significantly improved in the study arm compared with the control arm. Mean Headache Impact Test-6 score improved 12.4 points in the study arm after surgery (p < 0.05), while the mean postoperative score worsened by 5 points in the control arm, but this was not significant (p = 0.48). Subjectively, 25 of 28 study patients, and 4 of 19 control patients noticed at least some improvement in headache symptoms after surgery. Correction of visually significant upper eyelid position may improve chronic headache symptoms.


Gut ischemia and mesenteric synthesis of inflammatory cytokines after hemorrhagic or endotoxic shock.
The intestine plays a major role in the pathophysiology of multiorgan failure. Although the systemic inflammatory response might be induced by endotoxin released through bacterial translocation, other factors such as intestinal ischemia might be implicated. We investigated the relationship between intestinal ischemia-reperfusion and cytokine release in rat models of hemorrhagic or endotoxic shock. Plasma levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), lactate, and endotoxin, as well as macrophage TNF-alpha and IL-6 mRNA expression, were assessed at the end of shock and resuscitation. Hemodynamic changes and lactate levels suggested the presence of intestinal ischemia in both models. Mesenteric levels of TNF-alpha and IL-6 were increased by hemorrhage and further increased after saline resuscitation. Similar results were obtained with mRNA cytokine gene expression in macrophages. Endotoxin was not detectable in the hemorrhagic group. Endotoxic shock also increased production of cytokines, which, in contrast to hemorrhage, was not further increased by resuscitation. These results suggest that intestinal ischemia-reperfusion upon hemorrhage and resuscitation may be a major trigger for cytokine gene expression in the absence of endotoxin.


The h subunit of eIF3 promotes reinitiation competence during translation of mRNAs harboring upstream open reading frames.
Upstream open reading frames (uORFs) are protein coding elements in the 5' leader of messenger RNAs. uORFs generally inhibit translation of the main ORF because ribosomes that perform translation elongation suffer either permanent or conditional loss of reinitiation competence. After conditional loss, reinitiation competence may be regained by, at the minimum, reacquisition of a fresh methionyl-tRNA. The conserved h subunit of Arabidopsis eukaryotic initiation factor 3 (eIF3) mitigates the inhibitory effects of certain uORFs. Here, we define more precisely how this occurs, by combining gene expression data from mutated 5' leaders of Arabidopsis AtbZip11 (At4g34590) and yeast GCN4 with a computational model of translation initiation in wild-type and eif3h mutant plants. Of the four phylogenetically conserved uORFs in AtbZip11, three are inhibitory to translation, while one is anti-inhibitory. The mutation in eIF3h has no major effect on uORF start codon recognition. Instead, eIF3h supports efficient reinitiation after uORF translation. Modeling suggested that the permanent loss of reinitiation competence during uORF translation occurs at a faster rate in the mutant than in the wild type. Thus, eIF3h ensures that a fraction of uORF-translating ribosomes retain their competence to resume scanning. Experiments using the yeast GCN4 leader provided no evidence that eIF3h fosters tRNA reaquisition. Together, these results attribute a specific molecular function in translation initiation to an individual eIF3 subunit in a multicellular eukaryote.


[Coupling and rec-independence of the processes of replication and transposition of Pseudomonas aeruginosa phage D3112. The effect of the phage genes controlling the replication of DNA of D3112].
D3112 phage was shown to replicate via the process of coupled replication--transposition: the phage DNA is not excised from the chromosome after prophage induction and new phage copies insert into many different sites. The transposition is controlled by two D3112 early genes--A (mapped in the 1.5-3 kbp region) and B (3-4.5 kbp), and requires intact attL site (involvement of the phage right end attR not studied). D3112 is capable to transpose RP4 plasmid into the chromosome; both the D3112 and RP4 transpositions are rec-independent. The product of the early C gene which is not required for D3112 transposition has pleiotropic effect on the development of D3112 and is necessary for the process of D3112 DNA excision from the chromosome, for cell lysis as well as for mature phage production. We suggest that this gene is responsible for positive regulation of D3112 late genes expression, similar to the C gene of Mu phage or Q gene of lambda. Mutations in four D3112 late genes ts25, ts35, ts73 and ts110 do not affect transposition or excision processes. No detectable (less than 0.02 copies per cell) amount of linear or circular D3112 DNA is formed during the replication--transposition. Hence, in the course of replication and transposition processes D3112 genome has its ends permanently bound covalently to the chromosome. The excision of the D3112 DNA takes place at late stages.


Probable Mechanisms of Doxorubicin Antitumor Activity Enhancement by Ginsenoside Rh2.
Ginsenoside Rh2 increases the efficacy of doxorubicin (DOX) treatment in murine models of solid and ascites Ehrlich's adenocarcinoma. In a solid tumor model (treatment commencing 7 days after inoculation), DOX + Rh2 co-treatment was significantly more efficacious than DOX alone. If treatment was started 24 h after inoculation, the inhibition of tumor growth of a solid tumor for the DOX + Rh2 co-treatment group was complete. Furthermore, survival in the ascites model was dramatically higher for the DOX + Rh2 co-treatment group than for DOX alone. Mechanisms underlying the combined DOX and Rh2 effects were studied in primary Ehrlich's adenocarcinoma-derived cells and healthy mice's splenocytes. Despite the previously established Rh2 pro-oxidant activity, DOX + Rh2 co-treatment revealed no increase in ROS compared to DOX treatment alone. However, DOX + Rh2 treatment was more effective in suppressing Ehrlich adenocarcinoma cell adhesion than either treatment alone. We hypothesize that the benefits of DOX + Rh2 combination treatment are due to the suppression of tumor cell attachment/invasion that might be effective in preventing metastatic spread of tumor cells. Ginsenoside Rh2 was found to be a modest activator in a Neh2-luc reporter assay, suggesting that Rh2 can activate the Nrf2-driven antioxidant program. Rh2-induced direct activation of Nrf2 might provide additional benefits by minimizing DOX toxicity towards non-cancerous cells.


Microbial etiologies in community acquired pneumonia (CAP).
The objective of the study was determination of bacterial etiologic factors, including antibiotic atypical pathogens, of community acquired pneumonia (CAP) in adults and of sensitivity of isolated strains. The examined group comprised 50 patients with clinical and x-ray image of pneumonia. Patients' expectoration sputum was analyzed. Amongst all isolated bacteria, the most frequent were Staphylococcus aureus - 17.9%, Haemophilus parainfluenzae - 12.5% and H. influenzae - 8.9%. Identification of Corynebacterium pseudodiphtheriticum in 8.9% of CAP cases drew our particular attention. Staphylococcus aureus was the least antibiotic sensitive microorganism. In the majority of patients (26: 52%), serologic markers of chlamydial infection were determined. Pneumonia often results from mixed typical and atypical flora infection. High percentage of atypical pathogens in the examined material suggests the necessity to administer intracellularly acting antibiotics.


Stroke suggests increased risk of dementia.
Stroke is a major cause of disability in the elderly and considerably increases the risk of dementia, which is another important source of disability. This population-based study aimed to examine the risk of dementia in patients with stroke compared with non-stroke cases with similar comorbidities. Using the Taiwan National Health Insurance databank covering the period 2001-2007, this retrospective cohort study evaluated the risk of dementia in 10,884 patients with first stroke who had no history of dementia. In this study, we performed a 1:5 case-control matched analysis, in which cases were matched to controls based on their estimated propensity scores, which were estimated with demographics and associated risk factors. This approach reduced selection bias. Cox proportional hazards regression analysis was then used to estimate the risk of dementia in stroke patients. During the 5-year follow-up period, 1,487 (13.74%) stroke and 1,402 (2.59%) non-stroke patients suffered dementia. Stroke was independently associated with a 6.09 (95% confidence interval [CI], 5.66 to 6.55) times greater risk of dementia 5 years after stroke. Older age was associated with a higher incidence of dementia after stroke. Each stroke type had different impacts on the occurrence of dementia. The hazard ratio of dementia among hemorrhagic stroke patients was much higher than those of ischemic stroke and controls. The findings of this study suggest that stroke confers an increased risk of dementia, especially in the elderly and in patients with hemorrhagic stroke. We advocate the need for close observation and enhanced health education programs to benefit patients with stroke.


Long Exercise Test in the Interattack Period of Periodic Paralysis: A Useful and Sensitive Diagnostic Tool.
The reference values of long exercise test are inconsistent in previous studies. Our research aims to determine the reference values and improve diagnostic efficiency of long exercise test in periodic paralysis. In this study, 100 healthy controls, 35 non-periodic paralysis patients, and 47 patients with periodic paralysis were recruited. The procedures of long exercise test were conducted following McMannis(1986) method. The decrease rate in compound muscle action potential amplitudes was correlated with gender, and it was not influenced by age, height, weight, and exercise. After analyzing receiver operating characteristic curves, the compound muscle action potential amplitude decrease rate was stratified by sex. The mean area under the curve, sensitivity, and specificity were higher in male and female than all pooled subjects. The reference values of compound muscle action potential amplitude decrease rate were 46.8% in male and 26.9% in female, respectively. The long exercise test is a useful and sensitive diagnostic tool for primary periodic paralysis and thyrotoxic periodic paralysis. However, the reference values should be set by sex, which has better diagnostic performance, sensitivity, and specificity.