Rate-dependent prolongation of cardiac action potentials by a methanesulfonanilide class III antiarrhythmic agent. Specific block of rapidly activating delayed rectifier K+ current by dofetilide.
Class III antiarrhythmic agents act by selective prolongation of cardiac action potential duration (APD). Methanesulfonanilide class III agents (e.g., E-4031 and dofetilide) are extremely potent and lengthen action potentials in a "reverse" rate-dependent manner; i.e., effects are greater at low compared with high rates of stimulation. By using the whole-cell current-clamp technique in isolated guinea pig ventricular myocytes, APD was shortened by rapid pacing (244 +/- 16 msec at 30 pulses per minute, 166 +/- 8 msec at 240 pulses per minute; n = 8). Dofetilide (1 microM) prolonged APD more when cells were stimulated at the rate of 30 pulses per minute (44 +/- 10-msec increase) than at 240 pulses per minute (21 +/- 5-msec increase). We investigated the mechanism of APD prolongation using voltage-clamp techniques. Dofetilide selectively inhibited IKr (IC50, 31.5 nM), defined as the rapidly activating inward rectifying component of net delayed rectifier K+ current (IK), without effects on the larger but more slowly activating component of IK (IKs) or on the inward rectifier K+ current (IK1). To examine the rate-dependent effects of dofetilide on APD, trains of conditioning pulses to 0 mV (200-msec duration) were applied at either 30 or 240 pulses per minute to mimic the action potential experiments. Test pulses or ramps were given after the conditioning train to quantitate changes in IK1, IKr, or IKs. The magnitude of neither IK1 nor IKr was dependent on the rate of the preceding train of depolarizations. Sensitivity to block of IKr by dofetilide was rate independent.(ABSTRACT TRUNCATED AT 250 WORDS)


Change in biased thinking in a 10-session treatment for borderline personality disorder: further evidence of the motive-oriented therapeutic relationship.
Borderline Personality Disorder (BPD) is characterized by both maladaptive thinking and problematic schemas. Kramer and colleagues (2011) showed that using the motive-oriented therapeutic relationship (MOTR), based on the individualized understanding of the patient according to Plan Analysis (Caspar, 2007), can improve treatment outcomes for BPD. The present process-outcome pilot study aimed to examine the effects of the motive-oriented therapeutic relationship on the cognitive biases of patients with BPD. Change in biased cognitions in N=10 patients who were subject to MOTR was compared to that of N=10 patients who received psychiatric-psychodynamic treatment (Gunderson & Links, 2008). Results show a greater decrease in over-generalizations in patients who received MOTR, compared to the patients who received the psychiatric-psychodynamic treatment. These changes were related to outcome in various ways. These findings underline the importance of an individualized case formulation method in bringing about therapeutic change.


Is there an optimal respiratory reference position for self-navigated whole-heart coronary MR angiography?
To test the direct influence of the reference respiratory position on image quality for self-navigated whole-heart coronary MRI. Self-navigated whole-heart coronary MRI was performed in 11 healthy adult subjects. Respiratory motion was compensated for by using three different respiratory reference positions of the heart: end-inspiratory, end-expiratory, and the mean of the entire respiratory excursion. All datasets were reconstructed without motion compensation for comparison. Image quality was assessed in all reconstructions using signal-to-noise ratio (SNR) and contrst-to-noise ratio (CNR) measurements, as well as percentage vessel sharpness and visible length of the coronary arteries. While SNR and CNR remained close to constant in all reconstructions, a clear and significant improvement in vessel sharpness was identified in all motion corrected datasets with respect to their uncorrected counterpart (e.g., percentage sharpness of the proximal right coronary artery (RCA): 61.6 ± 8.2% for end-inspiration, 64.1 ± 10.7% for end-expiration, and 63.3 ± 7.0% for the mean respiratory position versus 55.0 ± 10.4 for the uncorrected datasets; P < 0.05). Among all motion corrected reconstructions, the use of an end-expiratory reference position most consistently provided the highest image quality. In particular, some of the improvements in vessel sharpness and length measured for end-expiration were statistically significant with respect to the reconstructions performed at end-inspiration (e.g., percentage sharpness of the proximal left anterior descending coronary: 58.2 ± 7.4% versus 55.8 ± 8.4%; P < 0.05; and visible length of the RCA: 125.7 ± 25.9 mm versus 114.4 ± 27.4 mm; P < 0.05). The use of end-expiration as a reference position for respiratory motion correction in free-breathing self-navigated whole heart coronary MRA significantly improves image quality. J


RFLP and physical mapping with an rDNA-specific endonuclease reveals that nucleolus organizer regions of Arabidopsis thaliana adjoin the telomeres on chromosomes 2 and 4.
Ribosomal RNA genes are organized in tandem arrays called nucleolus organizer regions (NORs). In a prior study, RFLP mapping on pulsed-field gels placed NOR2 at the northern tip of Arabidopsis thaliana chromosome 2. New polymorphisms have allowed the other NOR, NOR4, to be mapped to the northern tip of chromosome 4. To map NOR-associated loci, rDNA-specific cleavage by I-Ppol, an endonuclease with a 15 nucleotide recognition sequence involved in rDNA-homing of a mobile, self-splicing Group I intron in Physarum was exploited. I-Ppol digestion of A. thaliana genomic DNA liberated two telomere-containing fragments no larger than 13 kbp, and telomere polymorphisms identified using I-Ppol cosegregated with NOR2 and NOR4. Restriction mapping suggested that telomere-proximal rRNA genes are oriented with their 5' ends nearest the chromosome ends and their 3' ends nearest the centromere. This orientation was confirmed using the polymerase chain reaction to clone one of the telomere-rDNA junctions, most likely the junction on chromosome 4. The telomeric repeats join the terminal rRNA gene downstream of its promoter, suggesting that this first gene is inactive. Subtelomeric repetitive DNAs are absent at the telomere-rDNA junction. Localization of NOR2, NOR4 and their associated telomeres, TEL2N and TEL4N, respectively, provides end points for the genetic and physical maps of chromosomes 2 and 4.


Robust C-PdNi-CNF Sandwich-Structured Catalyst for Suzuki Reactions and Experimental Study on the Mechanism.
The stability of metal nanoparticles is one of the key issues for their catalytic applications. In this study, we fabricated a sandwich structure to protect the metal nanoparticles. A carbon layer was used to wrap the PdNi nanoparticles on the carbon fiber, and the whole preparation process was simple and green. Electron transfer occurs between the carbon layer and the metal nanoparticles, making the two more closely combined. As a catalyst for the Suzuki reaction, it exhibits highly efficient catalysis and excellent stability. The calculated TOF reaches 18 662 h-1. After nine cycles, there was almost no decrease in performance. Additionally, we found that the addition of iodobenzene into the chlorobenzene reaction system could significantly improve the chlorobenzene conversion, and we proved that the catalyst has fine activity and stability with a bright future in commercial applications. The possible catalytic mechanism of Suzuki reaction was proposed based on experimental results. This study provides a simple and green method to prepare encapsulated metal nanoparticle catalysts and gives a deep insight into Suzuki reaction.


Endogenous cannabinoid and opioid systems and their role in nicotine addiction.
Nicotine addiction is a complex behavioural alteration, in which many neuronal pathways and neurotransmitters are involved. For a long time, dopamine has been considered one of the most important neurotransmitters in mediating the rewarding effects of nicotine. In addition, a great amount of research suggests that the endogenous cannabinoid and opioid systems play an overall modulatory effect on the reward circuitry and participate in the addictive properties of most of the prototypical drugs of abuse. This review focuses on recent behavioural and biochemical data involving these systems in the different processes that contribute to tobacco addiction. A possible role for the endogenous cannabinoid and opioid systems in the rewarding properties of nicotine as well as in the development of nicotine physical dependence and relapse to nicotine-seeking behaviour will be examined. According to preclinical studies, clinical trials suggest that the manipulation of these systems with cannabinoid or opioid antagonists could be a potential therapeutical strategy for treating nicotine addiction.


Liver regeneration on chicken chorioallantoic membrane.
To explore how the liver regenerates, liver pieces from 15-day-old chicken embryos were grafted onto the chorioallantoic membrane (CAM) of 9-day-old chicken embryos and cultured for 11 days at the longest. The cultured liver pieces were examined histologically. The liver implants were gradually engulfed into the CAM and underwent necrosis of hepatocytes, except in their peripheral areas, during the first 1-4 days after grafting. Surviving cells in the peripheral areas began to proliferate 4 days after grafting. Thereafter, the cells were assembled into normal liver tissues and represented almost all the areas of the implants 9 days after grafting. Only after penetration of blood vessels from CAM did the liver implants enter a phase of rapid regeneration to form well-organized liver tissues. At the early stage of regeneration, the cells at the peripheral areas did not produce albumin, but reproduced it in the regenerated liver tissues, implying that hepatocytes restored their functions that were temporarily lost in the process of regeneration. Thus, we concluded that the liver pieces from 15-day-old chicken embryos had the ability to form normal liver tissues on CAM and that the blood supply played an important role in liver regeneration.


Therapeutic approach of primary bone tumours by bisphosphonates.
Bone tumours can be dissociated in two main categories: i) primary bone tumours (benign or malignant) including mainly osteosarcoma and other sarcomas.ii)and giant cell tumour and bone metastases originate from others cancer (Breast, prostate, kidney cancer, etc). These tumours are able to destroy or/and induce a new calcified matrix. However, the first step of bone tumour development is associated with an induction of bone resorption and the establishment of a vicious cycle between the osteoclasts and the tumour growth. Indeed, bone resorption contributes to the pathogenesis of bone tumour by the release of cytokines (IL6, TNFα) which govern the bone tumour's development and which are trapped into the bone matrix. Bisphosphonates (BPs) are chemical compounds of P-C-P structure with a high affinity for bone hydroxyapatite crystals. Thus, they have been used as a carrier for radio nucleotides to develop novel approaches of bone imaging. BPs exert also indirect anti-tumour activities in vivo. Indeed, BPs directly interfere with the bone microenvironment and target osteoclasts, endothelial cells and immune cells (tumour-associated macrophages, γ9δ2 T cells). BPs induce tumour cell death in vitro and same activity is suspected in vivo. The present review summarizes the mechanisms of actions of BPs as well as their clinical interests in bone primary tumours.


Purification and characterization of a recombinant G-protein-coupled receptor, Saccharomyces cerevisiae Ste2p, transiently expressed in HEK293 EBNA1 cells.
The production of milligram quantities of purified, active, folded membrane protein from heterologous expression systems remains a general challenge due to intrinsically low expression levels, misfolding, and instability. Here we report the overexpression and purification of milligram quantities of functional Saccharomyces cerevisiae G-protein-coupled receptor, Ste2p, from transiently transfected human embryonic kidney 293 EBNA1 cells. Fluorescent microscopy indicates localization of Ste2p-GFP and Fc-Ste2p-GFP fusion receptors to the cell membrane. Up to 2 mg (approximately 10 pmol/million cells) of the Fc-Ste2p-GFP fusion and 1 mg of a Ste2p-Strep-TagII/(His)8-tagged version were purified per liter of culture following protein A-Sepharose and Talon metal affinity chromatography, respectively. Two distinct fluorescent labels, the hydrophobic 7-(diethylamino)-3-(4'-maleimidylphenyl)-4-methylcoumarin (CPM) and the more hydrophilic fluorescein-5-maleimide (FM), were individually attached to the C-terminus of the alpha-mating factor ligand by addition of a reactive cysteine residue to produce active fluorescent pheromones. In vitro fluorescent ligand binding assays demonstrated that a high percentage of the recombinant purified receptor is correctly folded and able to bind ligand. KD values of 34 +/- 3 and 300 +/- 20 nM were observed respectively for the CPM- and FM-labeled ligands. These results combined with blue-shifted emission peaks and loss of fluorescent quenching observed for both fluorescent-labeled Cys alpha-factors when bound to receptor support a model in which the C-terminus of the ligand is packed in a hydrophobic pocket at the interface between the transmembrane and extracellular loop domains. Overall, we present an efficient system for recombinant production of milligram quantities of purified Ste2p in a biologically active form with applications to future structure and functional studies.


Brainstem Associated Somatosensory Evoked Potentials and Response to Vagus Nerve Stimulation: An Investigation of the Vagus Afferent Network.
Despite decades of clinical usage, selection of patients with drug resistant epilepsy who are most likely to benefit from vagus nerve stimulation (VNS) remains a challenge. The mechanism of action of VNS is dependent upon afferent brainstem circuitry, which comprises a critical component of the Vagus Afferent Network (VagAN). To evaluate the association between brainstem afferent circuitry and seizure response, we retrospectively collected intraoperative data from sub-cortical recordings of somatosensory evoked potentials (SSEP) in 7 children with focal drug resistant epilepsy who had failed epilepsy surgery and subsequently underwent VNS. Using multivariate linear regression, we demonstrate a robust negative association between SSEP amplitude (p < 0.01), and seizure reduction. There was no association between SSEP latency and seizure outcomes. Our findings provide novel insights into the mechanism of VNS and inform our understanding of the importance of brainstem afferent circuitry within the VagAN for seizure responsiveness following VNS.


Abdominal cutaneous nerve entrapment syndrome (ACNES) in a patient with a pain syndrome previously assumed to be of psychiatric origin.
A 21-year-old female patient with chronic abdominal pain was referred to a psychiatric outpatient clinic after gastroenterological and gynaecological pathogeneses had been excluded and a treatment with an antidepressant had had no beneficial effects. The mental state examination, however, revealed no psychopathology whatsoever. The patient was injected with a local anaesthetic loco dolenti which resulted in immediate pain relief. She was diagnosed with abdominal cutaneous nerve entrapment syndrome (ACNES); no psychiatric diagnosis was given.


Pyrrolizidine alkaloids in Pulmonaria obscura.
Leaves, roots and rhizomes, and inflorescenses of Pulmonaria obscura from two localities and of two different growth forms within one site were analyzed by GC-MS for their content in pyrrolizidine alkaloids (PAs). In roots and rhizomes PAs of the lycopsamine type typical for the Boraginaceae could be detected, e. g. intermedine, lycopsamine, and their O(7)-derivatives. The total PA concentrations in roots and rhizomes lay between 0.026 and 0.158 mg/g dry weight. In leaves and inflorescenses, on the other hand, only trace amounts of PAs (below 0.4 ng/mg dry weight) could be detected. No significant differences in total concentration of PAs could be found between the two sites.


Recovery of apparent diffusion coefficient after ischemia-induced spreading depression relates to cerebral perfusion gradient.
Transient decreases of the apparent diffusion coefficient (ADC) of water as measured by fast diffusion-weighted imaging (DWI) in the ischemic border zone are thought to reflect cellular swelling associated with spreading depression. DWI and dynamic contrast-enhanced MRI were applied to study the characteristics of spreading depression and the correlation between ADC recovery time and tissue perfusion in focal ischemia. Serial DWI was performed during remote middle cerebral artery occlusion in rats (n = 5) with an echo-planar imaging technique. ADC maps were calculated and ADC values displayed as a function of time in user-defined regions of interest with a time resolution of 12 to 16 seconds. Dynamic contrast-enhanced MRI was performed for qualitative correlation of ADC changes with tissue perfusion. Recovery time of transient ADC decreases correlated with the degree of the perfusion deficit (r = .81, P < .001). Slowly recovering ADC declines were found close to the ischemic core and correlated with severe perfusion deficit, while short-lasting ADC declines were typically found in moderately malperfused or normal tissue. Transient ADC decreases originated in the subcortical and cortical ischemic border zones and propagated along the cortex with a velocity of 2.9 +/- 0.9 mm/min. The variation in the recovery time of transient ADC decreases in the ischemic periphery reflects the gradient of the tissue perfusion. Severely delayed recovery time after spreading depression is thought to represent the ischemic penumbra.


Thermodynamic analysis of beta-hairpin-forming peptides from the thermal dependence of (1)H NMR chemical shifts.
The temperature dependence of the (1)H chemical shifts of six designed peptides previously shown to adopt beta-hairpin structures in aqueous solution has been analyzed in terms of two-state (beta-hairpin left arrow over right arrow coil) equilibrium. The stability of the beta-hairpins formed by these peptides, as derived from their T(m) (midpoint transition temperature) values, parallels in general their ability to adopt those structures as deduced from independent NMR parameters: NOEs, Deltadelta(C)(alpha)(H), Deltadelta(C)(alpha), and Deltadelta(C)(beta) values. The observed T(m) values are dependent on the particular position within the beta-hairpin that is probed, indicating that their folding to a beta-hairpin conformation deviates from a "true" two-state transition. To obtain individual T(m) values for each hairpin region in each peptide, a simplified model of a successive uncoupled two-state equilibrium covering the entire process has been applied. The distribution of T(m) values obtained for the different beta-hairpin regions (turn, strands, backbone, side chains) in the six analyzed peptides reveals a similar pattern. A model for beta-hairpin folding is proposed on the basis of this pattern and the reasonable assumption that regions showing higher T(m) values are the last ones to unfold and, presumably, the first to form. With this assumption, the analysis suggests that turn formation is the first event in beta-hairpin folding. This is consistent with previous results on the essential role of the turn sequence in beta-hairpin folding.


Electron Transport in Low Dimensional Solids: A Surface Chemistry Perspective.
Electron transport is a fundamental process that controls the intrinsic chemical and physical properties of solid materials. The surface phase becomes dominant when downsized dimensionality into cluster scale in nanomaterials, and surface chemistry plays more and more important role in regulating electron transport. During past decades, varieties of chemical approaches have been developed to modify the surface of low dimensional solids, substantially providing versatile perspectives on engineering electron transport. In this Perspective, we focus on recent researches concerning surface chemical modification strategies, such as surface molecular adsorption, atomic incorporation, defect engineering and spin scattering to engineer electron transport of typical one-/two-dimensional systems. Under the framework of Drude's transport model, we highlight the core role of micro degrees of freedom, i.e., charge, lattice, and spin, in molecular-level understanding and optimizing the regulation effect of surface chemistry. Finally, based on the discussion and current achievements of surface chemistry effect on electron transport of low dimensional solids, some personal perspectives on the future development are also presented.


Gastrointestinal stromal tumours: a clinico-radiologic review from a single centre in South India.
Gastrointestinal stromal tumours (GISTs) are rare tumours but are the commonest mesenchymal neoplasms in the gastrointestinal tract. To our knowledge, there is no large case series in Asian countries in which a clinico-radiological descriptive analysis of these tumours has been carried out. In this retrospective study, we analysed our experience of 70 patients with histopathologically proven GISTs, who were presurgically investigated by using CT, and describe the demography, anatomical distribution, imaging features and clinical course of the GIST. We found an unusually large predominance of males in our study, stomach and small bowel appeared to have been involved similarly and small bowel tumours had a higher rate of metastases. We also highlight some unusual CT features of these tumours that we encountered during the study, such as the presence of metastatic lymphadenopathy and satellite nodules, relapse in appendices epiploicae of the bowel, metachronous liposarcoma, adrenal and lung metastases, multiplicity of lesions and aneurysmal dilatation of the bowel. Two of our patients also had multiple neurofibromas, whose association with GIST has been seen in earlier reports. To the best of our knowledge, this article presents one of the largest series of articles on GISTs, to date, in Asian countries. We conclude with a differential diagnosis of GIST, with salient features distinguishing each entity.


A proposal for mitochondrial processing peptidase catalytic mechanism.
The reaction mechanism of the Mitochondrial Processing Peptidase enzyme (MPP) was investigated by using hybrid density functional theory. This enzyme removes the NH(2)-terminal targeting signals of nuclear-encoded mitochondrial protein precursors in the mitochondrial matrix. The catalytic process was studied using a model for the active site consisting of 161 atoms locating all the stationary points on the potential energy curve and determining the main energetic, structural, and electronic features that drive the catalysis. Despite the differences between the B3LYP and MPWB1K descriptions, it is possible hypothesize that the rate-limiting step of the reaction is most likely the nucleophilic attack of zinc-bound hydroxide to a carbonyl carbon of the substrate. The results allowed assignment of the proper roles to some active site residues in this mechanism.


Metalloproteinase inhibitor attenuates neointima formation and constrictive remodeling after angioplasty in rats: augmentative effect of alpha(v)beta(3) receptor blockade.
Release of matrix metalloproteinases (MMP) from smooth muscle and foam cells following arterial injury facilitates cell migration, neointimal hyperplasia, and vessel wall remodeling. Inhibition of MMP activity using the hydroxamate, zinc-chelating mimicers of collagen, Batimastat and Marimastat, has shown efficacy in reducing constrictive vascular remodeling 6 weeks after experimental angioplasty but not intimal hyperplasia. Vitronectin receptor (alpha(v)beta(3)) blockade interferes with binding of this integrin to MMP-2 and proteolyzed collagen, thereby reducing cell invasion. This study tests the effect of MMP inhibition, with and without vitronectin receptor (alpha(v)beta(3)) blockade, on neointima formation and arterial remodeling in a long-term model (up to 212 months) of balloon injury in vivo. Male Sabra rats were treated with Batimastat (BB-94, British Biotech Pharmaceuticals Ltd., 30 mg/kg, intraperitoneally) and/or the alpha(v)beta(3) receptor inhibiting RGD peptide, G-Pen-GRGDSPCA (GIBCO BRL, 0.1 micromol), administered as a perivascular gel to the common carotid artery after balloon injury. Animals were sacrificed 3, 14, 25, and 75 days (n=21, 23, 22, and 21) after injury. Animals treated with BB-94, peptide, or both had markedly increased absolute luminal area with markedly reduced luminal cross-sectional-area narrowing by neointima and intima-to-media area ratio at all time points except for 3 days after balloon injury versus non-treated, ballooned animals. Combined treatment was significantly more effective than either one alone. Constrictive remodeling, most marked 212 months after balloon injury, was prevented at this time point in all treated animals. The pattern of reduction in luminal narrowing, neointimal formation, and constrictive remodeling across treatment groups correlated very significantly with the reduction in tissue MMP activity as determined by zymography at 3 days. Confirmation of the efficacy of this strategy in larger animals should be the next step toward testing the applicability of this novel approach to the interventional setting.


Practical lessons from the first outbreaks: clinical presentation, obstacles, and management strategies for severe pandemic (pH1N1) 2009 influenza pneumonitis.
During the initial spring wave of novel influenza pH1N1 (2009), several North American cities experienced localized epidemics that served as a harbinger of the larger second Fall wave of infection. The city of Winnipeg, the capital of the province of Manitoba in central Canada, was one of the first in North America to deal with a rapid presentation of large numbers of patients requiring critical care services resulting from pandemic (pH1N1) 2009 influenza-associated respiratory failure. Mexico City, Orlando, FL, and Salt Lake City, UT, were other Northern Hemisphere sites of heavy disease activity during the spring wave of the pandemic. This article is written in a narrative format that allows the reader to understand the problems (both major and mundane, anticipated and unexpected) experienced by healthcare workers in these sites during this pandemic. Descriptions cover a range of issues and difficulties that caused significant stress to the operations of intensive care units in these cities. We hope to offer some insight into potential pitfalls and problems that may be experienced by other centers and provide some potential approaches to addressing these issues.


Information content and scalability in signal transduction network reconstruction formats.
One of the first steps towards holistic understanding of cellular networks is the integration of the available information in a human and machine readable format. This network reconstruction process is well established for metabolic networks, and numerous genome wide metabolic reconstructions are already available. Extending these strategies to signalling networks has proven difficult, primarily due to the combinatorial nature of regulatory modifications. The combinatorial nature of possible protein-protein interactions and post translational modifications affects both network size and the correspondence between the reconstructed network and the underlying empirical data. Here, we discuss different approaches to reconstruction of signal transduction networks. We divide the current approaches into topological, specific state based and reaction-contingency based, and discuss their different information content and scalability. The discussion focusses on graphical formats but the points are in general applicable also to mathematical models and databases. While the formats have complementary strengths especially for small networks, reaction-contingency based formats have a number of advantages in the light of global network reconstruction. In particular, they minimise the need for assumptions, maximise the congruence with empirical data, and scale efficiently with network size.


Phase transitions of adsorbed atoms on the surface of a carbon nanotube.
Phase transitions of adsorbed atoms and molecules on two-dimensional substrates are well explored, but similar transitions in the one-dimensional limit have been more difficult to study experimentally. Suspended carbon nanotubes can act as nanoscale resonators with remarkable electromechanical properties and the ability to detect adsorption at the level of single atoms. We used single-walled carbon nanotube resonators to study the phase behavior of adsorbed argon and krypton atoms as well as their coupling to the substrate electrons. By monitoring the resonance frequency in the presence of gases, we observed the formation of monolayers on the cylindrical surface, phase transitions within them, and simultaneous modification of the electrical conductance.


Australian nurse supervisors' styles and their perceptions of ethical dilemmas within health care.
To examine clinical nurse supervisors' styles, in terms of models, organization, focus on supervision and theories used in supervision, as well as their perceptions of ethical dilemmas within health care. The importance of clinical supervision in clinical practice has been reported. However, literature dealing with its implementation is rare. This study is a descriptive-correlational study. Data were collected by means of a questionnaire from two groups of nurse supervisors randomly selected from a university (n = 55) and a hospital (n = 31). Data were analysed using descriptive and inferential statistics. The models most frequently used were 'case-oriented' and 'decision-making oriented'. The nurse supervision was reported as being organized on a one-to-one basis. The focus of supervision was on 'patient problems' and 'cooperation in team'. The theory most commonly used was the theory of reflection. The supervisors' perceptions of moral dilemmas were related to 'decision making' and actions impacting on quality of care resulting from their supervisor styles. By correlating the supervisors' styles and their perceptions of ethical dilemmas, we conclude that it is important to contribute to further research on supervisors' styles in order to ensure successful supervision.


Overdiagnosis and overtreatment; how to deal with too much medicine.
The past few decades have seen medical science making rapid strides in the field of diagnostic technologies, thus making it possible to detect some conditions at a very early stage. An unwanted effect of these developments is overdiagnosis which occurs when a true abnormality is discovered, but detection of that abnormality and its treatment does not benefit the patient. Overdiagnosis is encountered as a result cancer screening, in genetic diseases and in some chronic conditions when disease definition is broadened. There is urgent need of development of evidence based decision-making tools for clinicians which will help patients understand the benefits and harms of different screening and treatment methods. And the treating physician has to play a important role to convince the patient that watchful waiting, for some of the lesions may be the best option available in some circumstances.


[Central lymphodissection in carcinoma of the thyroid gland].
Experience with central lymphodissection (CLD) was analyzed in 93 patients with primary thyroid cancer (pT1-4aN0-1a). Prophylactic central lymphodissection was fulfilled in 48 patients, therapeutic CLD in 45 patients. The size and spread of the primary thyroid tumor had no difference in both groups of patients. In patients of the 2nd group thyroidectomy was fulfilled more often (71.1%) than in the 1st group (58.3%).The frequency of postoperative transitory hypoparathyroidism after surgery on the thyroid gland and therapeutic CLD made up 13.3%. There were no recurrent metastases of cancer after CLD fulfilled during the last 3.5 years. All cancer metastases in the postoperative period were detected in lymph nodes of the neck (2-5 levels) and their prevalence was 8.9%.


[Extended lymphadenectomy in radical cystectomy: margins, scope, methodology].
Radical cystectomy with pelvic lymphadenectomy is a gold standard for treatment of muscle-invasive urinary bladder cancer. However, therapeutic and prognostic value of pelvic lymphadenectomy is still controversial. Recent studies have demonstrated a better prognosis after extended lymphadenectomy. A multicenter study was made to standardize an extended lymphadenectomy procedure. We examined prospectively the total number of lymph nodes removed from various sites, number and location of positive nodes and its relation to location of primary tumors in the urinary bladder. Thirty five radical cystectomies with extended lymphadenectomy were performed for the treatment of invasive bladder cancer in National Urology Center in 1999-2004. The margins of extended lymphadenectomy were: cranial level of a.mesenterica inferior; lateral--n.n. genitofemoralis; caudal-fossa obturatoria. A total amount of removed lymph nodes comprised 1081, mean 34.2 +/- 8.1 lymph node per patient (range from 10 to 58). Fourteen patients (40.0%) were node positive (69 nodes). According to N category: N1--6 patients; N2--7 patients. Six patients had lymph node metastases up to the aortic bifurcation. In 6 cases a positive node was found on the contralateral side. Thus, we recommend extended radical lymphadenectomy for all patients undergoing radical cystectomy for bladder cancer.


Inflammation in periodontal tissues in response to mechanical forces.
Orthodontic forces are known to produce mechanical damage and inflammatory reactions in the periodontium and dental pulp, as well as inflammatory mediators, e.g. prostaglandins, interleukin (IL)-1, IL-6, tumor necrosis factor alpha, and receptor activator of nuclear factor kappaB ligand, in the periodontal ligament (PDL) and dental pulp. We have studied the effects of aging on the production of inflammatory mediators in the PDL using in vitro and in vitro methods and found that aging of PDL tissues may be an important factor in the severity of periodontal disease through a higher production of inflammatory mediators in response to mechanical forces. Further, the levels of inflammatory mediators in gingival crevicular fluid, an osmotically mediated inflammatory exudates found in the gingival sulcus, have been shown to be significantly elevated during orthodontic treatment. In order to reduce inflammation, low-level laser therapy has been recently studied in vitro and in vitro by many investigators as a substitute for anti-inflammatory drugs. Clinical and experimental studies have shown that low-level laser irradiation reduces orthodontic post-adjustment inflammation. We believe that orthodontic forces (mechanical forces) may play an important role in periodontal inflammation and that low-level laser therapy may be useful for its inhibition.


Fluorination of mixed γ-alumina/γ-gallia xerogels with trifluoromethane: some effects on bulk and surface characteristics.
Interaction of single γ-Al2O3 and γ-Ga2O3, and mixed γ-Al2O3/γ-Ga2O3 xerogels with CHF3 at intermediate temperatures results in partial fluorination. Fluorinated oxides remain amorphous and retain a considerable part of the initial surface area; for the fluorinated Al-based materials surface areas in all cases exceed 100 m2 g-1. Lewis acidity of mixed oxides, either before or after fluorination, is strongly influenced by the presence of surface Ga3+ ions, mainly due to their strong preference to replace highly acidic Al3+ ions in tetrahedral positions. Ion replacement leads to the formation of acidic sites with lower strengths what is confirmed by the model catalytic reaction, isomerisation of CCl2FCClF2. XPS investigations indicate that fluorination of mixed oxides is accompanied by substantial surface reconstructions and preferential formation of Al-F based phases with Ga remaining mainly in O environments. Further segregation processes, such as slow crystallisation of Al(F,OH)3·nH2O phases, are probably promoted by water adsorption.


[Results of posterolateral spinal fusion on spondylolisthesis (author's transl)].
44 patients with spondylolisthesis (type I Meyerding) which had had posterolateral fusion could be followed up for an average of five years. The operation had been performed for low back pain with or without sciatica when conservative treatment was not successful. In 42 patients (95%) fusion was solid and in 35 patients (80%) the symptomes could be distinctly deminished. Posterolateral fusion is a successful treatment with few complications in spondylolisthesis which does not respond to conservative measures.


Potential Antioxidant Activity of Calcium and Selected Oxidative Stress Markers in Lead- and Cadmium-Exposed Workers.
Occupational lead (Pb) and cadmium (Cd) exposure occurs during processing and casting of nonferrous metals such as zinc. In contrast to Pb and Cd, Ca is essential for living organisms due to its important role in a multitude of functions, from cell signaling to bone growth. Pb and Cd exposure affects calcium metabolism in various ways. The aim of this study was to investigate the blood levels of Pb, Cd, and Ca and the levels of selected oxidative stress biomarkers in workers exposed to Pb and Cd. Population groups included 264 male employees in a lead-zinc smelter. The study population was divided into two subgroups based on the median of Ca serum level (2.42 mmol/l): the low-Ca-level group (L-Ca group) and the high-Ca-level group (H-Ca group). Ca level was significantly higher in the H-Ca group than in the L-Ca group due to the study design (by 26%). The level of zinc protoporphyrin (ZPP) was significantly higher in the L-Ca group than in the H-Ca group by 13%, while the blood lead levels (PbB) were similar in the examined groups. The level of cadmium (CdB) was significantly higher in the L-Ca group than in the H-Ca group by 33%. From oxidative stress markers in serum, only the levels of malondialdehyde (MDA) and ceruloplasmin (CER) were significantly higher in the L-Ca group than in the H-Ca group, by 12% and 4%, respectively. The correlation analysis showed negative correlations between Ca level and the levels of PbB, ZPP, CdB, and MDA. The presented results indicate that Ca level modulates the serum concentration of Cd and has an impact on Pb-induced impairment of heme synthesis. The higher Ca levels may lead to a decrease in the concentration of lipid peroxidation products. Moreover, serum calcium level seems to be able to modify the level of acute-phase proteins. Obtained results suggest that higher Ca level may be useful in reducing Cd level in occupationally exposed workers.


Sugar regulation of plastid interconversions in epicarp of citrus fruit.
Seasonal transformations between chloroplasts and chromoplasts, as measured by changes in chlorophyll content, in the epicarp of degreening and regreening Citrus sinensis (L.) Osbeck cv Valencia fruit closely parallelled the accumulation and later loss of soluble sugars. At any stage of development, reversing the relative soluble sugar content in the epicarp by culturing pericarp segments on agar media with low (15 millimolar) or high (150 millimolar) sucrose concentrations reversed the direction of change in chlorophyll content. Fruit of C. madurensis Lour., which mature year around and do not regreen, also accumulated soluble sugars in the pericarp as degreening was initiated.The epicarp of C. sinensis fruit accumulated nitrogen, but total nitrogen concentrations and amino acid concentrations changed little, during degreening and regreening of C. sinensis fruit. Cessation of nitrogen fertilization reduced the tendency of pericarp segments to regreen in vitro during subsequent years, but regreening tendency was restored by inclusion of KNO(3) in the media.It is concluded that chloroplasts become chromoplasts and citrus fruit degreen partially in response to the accumulation of sugars in the epicarp and that the reverse transformation accompanying regreening of certain citrus species occurs when accumulated sugars disappear. Change in nitrogen flux to the fruit is probably not a factor in regulating seasonal transformations, but an abundance of nitrogen in the epicarp diminishes the effects of high sugar concentrations in inducing transformation of chloroplasts to chromoplasts, thereby retarding degreening and promoting regreening.


Long-Term Effects of Chronic Buspirone during Adolescence Reduce the Adverse Influences of Neonatal Inflammatory Pain and Stress on Adaptive Behavior in Adult Male Rats.
Neonatal pain and stress induce long-term changes in pain sensitivity and behavior. Previously we found alterations in pain sensitivity in adolescent rats exposed to early-life adverse events. We tested whether these alterations have long-lasting effects and if those effects can be improved by the 5-hydroxytryptamine 1A (5-HT1A) receptor agonist buspirone injected chronically during the adolescent period. This study investigates: (1) effects of inflammatory pain (the injection of formalin into the pad of a hind paw) or stress (short maternal deprivation-isolation, MI), or their combination in 1-2-day-old rats on the adult basal pain, formalin-induced pain, anxiety and depression; (2) effects of adolescent buspirone in adult rats that experienced similar early-life insults. Changes in nociceptive thresholds were evaluated using the hot plate (HP) and formalin tests; levels of anxiety and depression were assessed with the elevated plus maze and forced swim tests respectively. Both neonatal painful and stressful treatments induced long-term alterations in the forced swim test. Other changes in adult behavioral responses were dependent on the type of neonatal treatment. There was a notable lack of long-term effects of the combination of early inflammatory pain and stress of MI on the pain responses, anxiety levels or on the effects of adolescent buspirone. This study provides the first evidence that chronic injection of buspirone in adolescent rats alters antinociceptive and anxiolytic effects limited to adult rats that showed behavioral alterations induced by early-life adverse treatments. These data highlight the role of 5-HT1A receptors in long-term effects of neonatal inflammatory pain and stress of short MI on adaptive behavior and possibility of correction of the pain and psychoemotional behavior that were altered by adverse pain/stress intervention using buspirone during critical adolescent period.


Utilizing the Consolidated Framework for Implementation Research to Explore Palliative Care Program Implementation for American Indian and Alaska Natives throughout the United States.
Background: A significant shortage of palliative care (PC) services exists for American Indian and Alaska Native people (AI/ANs) across the United States. Using an implementation science framework, we interviewed key individuals associated with AI/AN-focused PC programs to explore what is needed to develop and sustain such programs. Objectives: To identify facilitators of implementation and barriers to sustainability associated with the development of PC programs designed for AI/ANs across the United States. Methods: We interviewed 12 key individuals responsible for the implementation of AI/AN-focused PC services. The Consolidated Framework for Implementation Research (CFIR) guided data coding and interpretation of themes. Results: We identified nine themes that map to CFIR constructs. Facilitators of implementation include high tension for change and respecting cultural values. Barriers to program sustainability include a lack of administrative leadership support. Discussion: AI/AN-focused PC programs should be congruent with community needs. PC program developers should focus on sustainability well before initial implementation.


[Analysis of acid-base disturbances in the emergency department].
The analysis of acid-base disturbances contributes to the diagnostic work-up of critically ill patients. Most emergency departments are equipped with blood gas point-of-care analyzers that quantify within minutes pH, pCO2, standard bicarbonate, standard base excess, sodium and chloride levels. This article provides a pragmatic stepwise approach to the analysis of acid-base disturbances in the emergency department. Standard base excess is used to assess the adequacy of the secondary (compensatory) response. Calculation of the anion gap based on the actual bicarbonate is used to identify the coexistence of metabolic acidosis and metabolic alkalosis. The delta anion gap allows for the identification of measurement errors, such as falsely elevated lactate and chloride values, which in turn may provide diagnostic clues.


Short-term effects of organic amendments on properties of a Vertisol.
Application of organic waste products as amendments has been proposed as a management option whereby soil quality of Vertisols could be improved. An incubation experiment was, therefore, conducted for 4 weeks under controlled temperature conditions (30 degrees C) to identify those potential organic amendments that might improve the quality of a Vertisol. Twelve organic amendments were investigated: cotton gin trash from three sources, cattle manure from two sources, green waste compost, chicken manure from three sources including a commercial product, biosolids and two commercial liquefied vermicomposts. Except for the biosolids, no other organic amendments had any effect on soil microbial biomass and respiration. Compared with NO3-N levels in the control, there was a 50% decrease in soil amended with 10 t ha(-1) green waste compost (65 microg g(-1)). The three different types of chicken manures increased the NO3-N concentration from 75% (228 microg g(-1)) to 226% (424 microg g(- 1)) over the control. Approximate recovery of P added by the amendment as resin-extractable soil P was 53% for cattle manure and 39% for chicken manure. Application of cattle manure resulted in a 22% increase in soil-exchangeable K over levels found in control. Organic amendments application also resulted in a significant increase in exchangeable Na concentration. Some of the organic wastes, viz. cotton gin trash (10 t ha(-1)), cattle manure (10 t ha(-1)), biosolids (10 t ha(-1)) and composted chicken manure (3 t ha(-1)) have value as a source of nutrients to soil and hence showed potential to improve Vertisol properties.


Progenitor/stem cell fate determination: interactive dynamics of cell cycle and microvesicles.
We have shown that hematopoietic stem/progenitor cell phenotype and differentiative potential change throughout cell cycle. Lung-derived microvesicles (LDMVs) also change marrow cell phenotype by inducing them to express pulmonary epithelial cell-specific mRNA and protein. These changes are accentuated when microvesicles isolated from injured lung. We wish to determine if microvesicle-treated stem/progenitor cell phenotype is linked to cell cycle and to the injury status of the lung providing microvesicles. Lineage depleted, Sca-1+ (Lin-/Sca-1+) marrow isolated from mice were cultured with interleukin 3 (IL-3), IL-6, IL-11, and stem cell factor (cytokine-cultured cells), removed at hours zero (cell cycle phase G0/G1), 24 (late G1/early S), and 48 (late S/early G2/M), and cocultured with lung tissue, lung conditioned media (LCM), or LDMV from irradiated or nonirradiated mice. Alternatively, Lin-/Sca-1+ cells not exposed to exogenous cytokines were separated into G0/G1 and S/G2/M cell cycle phase populations by fluorescence-activated cell sorting (FACS) and used in coculture. Separately, LDMV from irradiated and nonirradiated mice were analyzed for the presence of adhesion proteins. Peak pulmonary epithelial cell-specific mRNA expression was seen in G0/G1 cytokine-cultured cells cocultured with irradiated lung and in late G1/early S cells cocultured with nonirradiated lung. The same pattern was seen in cytokine-cultured Lin-/Sca-1 cells cocultured with LCM and LDMV and when FACS-separated Lin-/Sca-1 cells unexposed to exogenous cytokines were used in coculture. Cells and LDMV expressed adhesion proteins whose levels differed based on cycle status (cells) or radiation injury (LDMV), suggesting a mechanism for microvesicle entry. These data demonstrate that microvesicle modification of progenitor/stem cells is influenced by cell cycle and the treatment of the originator lung tissue.


Detection of microembolic signals in patients with neuropsychiatric lupus erythematosus.
The pathogenesis of central nervous system involvement in systemic lupus erythematosus (SLE) is not completely understood. In this study, we investigated the association of microembolic signals (MES) with a variety of neuropsychiatric SLE manifestations and compared our results with those from SLE patients without neuropsychiatric lupus and normal controls. Fifty-three patients with SLE (45 females and 8 males), all fulfilling the revised classification criteria for SLE, and 50 control subjects (44 females and 6 males) were enrolled in this study. All SLE patients were assessed by neuropsychological examination, including various neuropsychiatric tests. Twenty-five patients with SLE were found to have at least one of the neuropsychiatric syndromes defined by The American College of Rheumatology. The mean MES count in patients with neuropsychiatric lupus was significantly higher than those without (5.4 +/- 1.1 vs. 0.3 +/- 0.8/h; p < 0.005). We found a positive correlation between higher mean MES counts and the presence of neuropsychiatric syndromes in SLE. The mean MES count in the whole group of SLE patients was also significantly higher than that in healthy controls. The mean MES count of SLE patients with antiphospholipid (aPL) antibody positivity was significantly higher than those without aPL antibodies (3.6 +/- 1.6 vs. 0.8 +/- 0.1/h; p < 0.005). In conclusion, the association of MES with neuropsychiatric lupus may support the possible contribution of MES to the complex pathophysiology of this syndrome. More importantly, detection of MES on transcranial Doppler monitoring might suggest a high risk of involvement of the central nervous system in SLE, and could be used as a diagnostic tool.


Does funeral preference influence clinical necropsy request outcome?
Attitudes towards necropsy have been shown to be more favourable amongst those relatives preferring cremation as a method of disposal compared to those with a preference for burial. In a two-year retrospective study, no significant relationship was found between funeral preferences (burial or cremation) and clinical necropsy request outcome when age, sex and religion were taken into account. Potential religious objections to necropsy were infrequent and cremation was found to have become the most popular method of disposing of the dead during a period when local clinical necropsy rates have continued to decline. Funeral preference is unlikely to have been a significant factor in the decline in clinical necropsy rates.


Postsynaptic dorsal column and cuneate correlations in the raccoon: a re-evaluation by parallel-cascade analysis.
In a previous study, we reported evidence for correlations between the firing of postsynaptic dorsal column (PSDC) neurons and cuneate neurons with overlapping receptive fields on the glabrous skin of the raccoon forepaw. The evidence was based on cross-correlation and frequency response analyses of spontaneously firing neurons. However, cross-correlation without white noise Gaussian analog inputs or Poisson distributed pulse train inputs is difficult to interpret because of the inherent convolution with the autocorrelation of the unknown input signals. While the data suggested positive correlations in the spinocuneate direction for most neuron pairs, we could not estimate the temporal characteristics of these putative connections. We have now re-analyzed these data using a parallel-cascade method to estimate the first- and second-order kernels of a Volterra series approximation to the spinocuneate system. This unbiased analysis suggests that a positive correlation occurs after about 5 ms, probably followed by a negative correlation at about 12 ms. Second-order kernels also had repeatable structure, indicating dual pathways with time separations of at least 10 ms.


Continuous oral N-acetylcysteine treatment and development of nitrate tolerance in patients with stable angina pectoris.
The presence of sulfhydryl (SH) groups appears to be fundamental to nitrate-induced vasodilation and N-acetylcysteine (NAC), a sulfhydryl (SH)-donor substance, potentiates hemodynamic responsiveness to nitrates. We investigated the effect of simultaneous administration of NAC and isosorbide dinitrate (ISDN) on development of nitrate tolerance. In a double-blind, randomized, placebo-controlled cross-over study, seven patients with stable angina pectoris were treated for two 8-day periods with ISDN (40 mg four times daily, q.i.d.) together with NAC (controlled release 600 mg q.i.d.) or matching placebo. Bicycle exercise tests were performed before treatment was started, 1 h after treatment was started, and at day 8. After 8-day treatment with ISDN + placebo, responses determined by exercise testing were diminished as compared with responses obtained during acute therapy and did not differ from baseline values, suggesting development of tolerance to ISDN. During treatment with ISDN + NAC, time to 1-mm ST depression was significantly prolonged (441 +/- 44 vs. 381 +/- 40 s, mean +/- SEM) and total ST segment depression significantly reduced (1.9 +/- 0.7 vs. 3.5 +/- 0.8 mm) as compared with baseline values. The reduction in ST segment depression was significantly more pronounced during ISDN + NAC (46%) as compared with ISDN + placebo (23%). Although exercise time and time to angina pectoris were unaffected. NAC augmented the antiischemic effects of ISDN as assessed by ECG. This finding may suggest that development of nitrate tolerance is modified by chronic oral high-dose NAC administration.


Poor writing, not specialized concepts, drives processing difficulty in legal language.
Despite their ever-increasing presence in everyday life, contracts remain notoriously inaccessible to laypeople. Why? Here, a corpus analysis (n ≈10 million words) revealed that contracts contain startlingly high proportions of certain difficult-to-process features-including low-frequency jargon, center-embedded clauses (leading to long-distance syntactic dependencies), passive voice structures, and non-standard capitalization-relative to nine other baseline genres of written and spoken English. Two experiments (N=184) further revealed that excerpts containing these features were recalled and comprehended at lower rates than excerpts without these features, even for experienced readers, and that center-embedded clauses inhibited recall more-so than other features. These findings (a) undermine the specialized concepts account of legal theory, according to which law is a system built upon expert knowledge of technical concepts; (b) suggest such processing difficulties result largely from working-memory limitations imposed by long-distance syntactic dependencies (i.e., poor writing) as opposed to a mere lack of specialized legal knowledge; and (c) suggest editing out problematic features of legal texts would be tractable and beneficial for society at-large.


Host-soluble galectin-1 promotes HIV-1 replication through a direct interaction with glycans of viral gp120 and host CD4.
Sexual transmission of HIV-1 requires virus adsorption to a target cell, typically a CD4(+) T lymphocyte residing in the lamina propria, beneath the epithelium. To escape the mucosal clearance system and reach its target cells, HIV-1 has evolved strategies to circumvent deleterious host factors. Galectin-1, a soluble lectin found in the underlayers of the epithelium, increases HIV-1 infectivity by accelerating its binding to susceptible cells. By comparison, galectin-3, a family member expressed by epithelial cells and part of the mucosal clearance system, does not perform similarly. We show here that galectin-1 directly binds to HIV-1 in a β-galactoside-dependent fashion through recognition of clusters of N-linked glycans on the viral envelope gp120. Unexpectedly, this preferential binding of galectin-1 does not rely on the primary sequence of any particular glycans. Instead, glycan clustering arising from the tertiary structure of gp120 hinders its binding by galectin-3. Increased polyvalency of a specific ligand epitope is a common strategy for glycans to increase their avidity for lectins. In this peculiar occurrence, glycan clustering is instead exploited to prevent binding of gp120 by galectin-3, which would lead to a biological dead-end for the virus. Our data also suggest that galectin-1 binds preferentially to CD4, the host receptor for gp120. Together, these results suggest that HIV-1 exploits galectin-1 to enhance gp120-CD4 interactions, thereby promoting virus attachment and infection events. Since viral adhesion is a rate-limiting step for HIV-1 entry, modulation of the gp120 interaction with galectin-1 could thus represent a novel approach for the prevention of HIV-1 transmission.


A rare cause of severe recurrent abdominal pain.
Recurrent abdominal pain may be enigmatic to solve. We report three cases, presenting after a protracted history of severe recurrent bouts of epigastric pain, impeding daily activities. In all cases, serology for strongyloidiasis stercoralis proved positive. In one case, stool examination was positive for larvae. Treatment with ivermectin or albendazole resulted in complete resolution of abdominal symptoms without recurrence in all three cases.


A natural intergenotypic recombinant of hepatitis C virus identified in St. Petersburg.
Hepatitis C virus (HCV) evolution is thought to proceed by mutations within the six genotypes. Here, we report on a viable spontaneous HCV recombinant and we show that recombination may play a role in the evolution of this virus. Previously, 149 HCV strains from St. Petersburg had been subtyped by limited sequencing within the NS5B region. In the present study, the core regions of 41 of these strains were sequenced to investigate the concordance of HCV genotyping for these two genomic regions. Two phylogenetically related HCV strains were found to belong to different subtypes, 2k and 1b, according to sequence analysis of the 5' untranslated region (5'UTR)-core and the NS5B regions, respectively. By sequencing of the E2-p7-NS2 region, the crossover point was mapped within the NS2 region, probably between positions 3175 and 3176 (according to the numbering system for strain pj6CF). Sequencing of the 5'UTR-core regions of four other HCV strains, phylogenetically related to the above-mentioned two strains (based on analysis within the NS5B region), revealed that these four strains were also recombinants. Since a nonrecombinant 2k strain was found in St. Petersburg, the recombination may have taken place there around a decade ago. Since the frequency of this recombinant is now high enough to allow the detection of the recombinant in a fraction of the city's population, it seems to be actively spreading there. The reported recombinant is tentatively designated RF1-2k/1b, in agreement with the nomenclature used for HIV recombinants. Recombination between HCV genotypes must now be considered in the classification, laboratory diagnosis, and treatment of HCV infection.


Molecular cytogenetic evidence of rearrangements on the Y chromosome of the threespine stickleback fish.
To identify the processes shaping vertebrate sex chromosomes during the early stages of their evolution, it is necessary to study systems in which genetic sex determination was recently acquired. Previous cytogenetic studies suggested that threespine stickleback fish (Gasterosteus aculeatus) do not have a heteromorphic sex chromosome pair, although recent genetic studies found evidence of an XY genetic sex-determination system. Using fluorescence in situ hybridization (FISH), we report that the threespine stickleback Y chromosome is heteromorphic and has suffered both inversions and deletion. Using the FISH data, we reconstruct the rearrangements that have led to the current physical state of the threespine stickleback Y chromosome. These data demonstrate that the threespine Y is more degenerate than previously thought, suggesting that the process of sex chromosome evolution can occur rapidly following acquisition of a sex-determining region.


Diagnosis Dilemma of Angioimmunoblastic T-Cell Lymphoma in Tuberculosis Endemic Region.
Angioimmunoblastic T-cell lymphoma (AITL) is a rare hematologic malignancy recognized in the WHO 2016 classification as a clinical and histological entity. It is a very poorly described disease in Africa due to its rarity and diagnostic difficulties, particularly differential diagnosis with tuberculosis. Here, we report a 57-year-old man who presented with fever, weight loss, and lymphadenopathies. The diagnosis of tuberculosis was carried out based on lymph node fine needle aspiration showing the image of tuberculous adenitis and CT images in favor of necrotic lymphadenopathies. The presence of autoantibodies and the failure of tuberculosis treatment led us to perform a biopsy with immunostaining that confirmed pathological features of AITL. The patient was treated by CHOP-based chemotherapy, and complete remission was achieved. This case highlights the difficulty of recognizing AITL and the importance of considering other potential differential diagnoses of tuberculosis in the endemic region.


Tourniquet application during TKA did not affect the accuracy of implant positioning: a randomized clinical trial.
The accuracy of the identification of anatomical landmarks with versus without tourniquet application and its effect on implant positioning remained unknown. Therefore, the hypothesis of the present study was that tourniquet application did not affect the accuracy of the reconstruction of the mechanical leg alignment, the joint line level, and the patellar height. The prospective randomized monocentric trial (Clinical-Trials.gov NCT02475603) included a total of 86 patients scheduled to undergo primary TKA. The patients were allocated to receive TKA with (Group A, n = 43) or without tourniquet (Group B, n = 43). The mechanical leg alignment, the joint line level (modified Kawamura), and the patellar height (Plateau-patella angle, Insall Salvati index, and modified Insall Salvati index) were measured pre- and postoperatively on standardized calibrated digital radiographs. Mean, SEM, median, range, and p value were calculated for each parameter. There was no statistical difference between the groups with regard to demographics, preoperative deformity, implant design, and surgical technique (n.s.). The mechanical leg alignment, the joint line level, and the patellar height revealed, in both groups, similar results pre- and postoperatively (n.s.). The mechanical leg alignment, the joint line level, and the patellar height could be accurately reconstructed with and without tourniquet use. With respect to clinically relevant surrogate parameters of implant positioning, TKA can safely be performed without a tourniquet. Available data do not support a routine use of tourniquet during TKA and might justify a change of the clinical pathway. Level I.


Differential serum levels of eosinophilic eotaxins in primary sclerosing cholangitis, primary biliary cirrhosis, and autoimmune hepatitis.
To investigate pathogenic mechanisms of primary sclerosing cholangitis (PSC), primary biliary cirrhosis (PBC), and autoimmune hepatitis (AIH), serum levels of 26 chemokines and cytokines were determined and compared with patients with chronic hepatitis C or in healthy controls. The chemokine eotaxin-3 (E3; CCL26), which recruits eosinophils to sites of inflammation, was found to be highly elevated in all PSC, PBC, and AIH patients compared with HCV patients and healthy controls. Eotaxin-1 (E1; CCL11), another eosinophil-specific chemokine, was elevated in PSC but reduced in PBC and AIH, while the macrophage-derived chemokine (MDC; CCL22) was lower in all PSC, PBC, and AIH patients compared with HCV patients and controls. By incorporating levels of the interleukin (IL)-15 into a diagnostic algorithm, PSC, PBC, and AIH patients could each be differentiated with good sensitivity and specificity. These findings represent the first study to compare the level of serum cytokine/chemokine levels among these related autoimmune-like liver diseases. Furthermore, our data indicate that the measurement of serum E3, E1, CCL22, and IL-15 levels can aid in the diagnosis of these clinically challenging diseases and shed light on the potential pathogenic mechanisms underlying these diseases. By suggesting a potential role for an allergic phenomenon involving eosinophils, which may define them as liver-specific allergic diseases, this may open up potential new therapeutic avenues by abrogating the action of these disease-associated immune modulators.


Purification and characterization of two active derivatives of recombinant YplA, a secreted phospholipase from Yersinia entercolitica.
The virulence-associated phospholipase of Yersinia enterocolitica (YplA), which is secreted by a flagellar type III secretion system, was cloned and purified for structure-function analysis using a His(6)-tag expression system. Two versions of YplA have been proposed on the basis of two potential initiating methionine residues. The longer derivative possesses 59 additional amino acids at its N-terminus and appears to represent the native form of YplA; however, the shorter recombinant protein possesses enhanced activity in vitro. Both recombinant YplA derivatives are highly active as type-A(2) phospholipases and possess similar physical properties. Based on type III secretion substrates from other gram-negative bacteria, the N-terminus of YplA is probably required as a secretion signal; however, differences in the time-based activity of these two recombinant enzymes, the N-terminus of YplA may also have a regulatory function.


Suppression of inducible nitric oxide synthase expression and nitric oxide production by macrolide antibiotics in sulfur mustard-exposed airway epithelial cells.
Sulfur mustard, a vesicant chemical warfare agent, causes airway injury due to massive release of destructive enzymes and mediators of inflammation. Nitric oxide plays an important yet controversial role in inflammation. An impressive number of reports suggest that excessive amount of nitric oxide may promote inflammation-induced cell injury and death. Overproduction of nitric oxide is catalysed by up-regulated expression of the inducible isoform of nitric oxide synthase (iNOS). In this study, we used quantum dot-mediated immunocytochemistry to analyse iNOS expression and flow cytometry to analyse the intracellular nitric oxide production in sulfur mustard-exposed normal human small airway epithelial cells and bronchial/tracheal epithelial cells and studied the effect of four US Food and Drug Administration-approved macrolide antibiotics, namely, azithromycin, clarithromycin, erythromycin and roxithromycin. Exposure to 100 microM sulfur mustard significantly up-regulated iNOS expression and resulted in overproduction of nitric oxide in these cells. Addition of macrolide antibiotics to 100 microM in the medium reduced both iNOS expression and nitric oxide production to near normal level. Thus, the current study provides in vitro evidence of the immunomodulatory effects of macrolide antibiotics in sulfur mustard-exposed airway epithelial cells. These results suggest that macrolide antibiotics may serve as potential vesicant respiratory therapeutics through mechanisms independent of their antibacterial activity.


Influence of retinoblastoma-related gene silencing on the initiation of DNA replication by African cassava mosaic virus Rep in cells of mature leaves in Nicotiana benthamiana plants.
Geminiviruses mainly infect terminally differentiated tissues and cells in plants. They need to reprogramme host cellular machinery for DNA replication. This process is thought to be mediated by inactivation of cell-cycle repressor proteins and by induction of host DNA synthesis protein expression through actions of the geminviral replication initiator protein (Rep). Exploiting a Nicotiana benthamiana pOri2 line, which is transformed with a transgene consisting of a direct repeat of the African cassava mosaic virus (ACMV)-replication origin (Ori) flanking a non-viral DNA region, and virus-induced RNA silencing (VIGS), the impact of host gene expression on replication of the ACMV-derived replicon was investigated. The ACMV Rep trans-replicated the viral episomal replicon in leaves of young but not older pOri2 plants. Upon VIGS-mediated down-regulation of N. benthamiana NbRBR1, the retinoblastoma-related protein gene coding for a negative cell-cycle suppressor, recovered the ability of ACMV Rep for trans DNA replication, whereas the silencing of NbPCNA coding for the sliding clamp of DNA polymerase had no effect. These results suggest that the cellular machinery for DNA replication in differentiated tissues of older leaves cannot be reprogrammed by Rep alone but may need other uncharacterised viral and plant factors.


Twist1 Influences the Expression of Leading Members of the IL-17 Signaling Pathway in HER2-Positive Breast Cancer Cells.
Breast cancer (BC) is a heterogeneous disease composed of multiple subtypes with different molecular characteristics and clinical outcomes. The metastatic process in BC depends on the transcription factors (TFs) related to epithelial-mesenchymal transition (EMT), including the master regulator Twist1. However, its role beyond EMT in BC subtypes remains unclear. Our study aimed to investigate the role of Twist1, beyond EMT, in the molecular subtypes of BC. In patients, we observed the overexpression of TWIST1 in the HER2+ group. The silencing of TWIST1 in HER2+ BC cells resulted in the upregulation of 138 genes and the downregulation of 174 genes compared to control cells in a microarray assay. In silico analysis revealed correlations between Twist1 and important biological processes such as the Th17-mediated immune response, suggesting that Twist1 could be relevant for IL-17 signaling in HER2+ BC. IL-17 signaling was then examined, and it was shown that TWIST1 knockdown caused the downregulation of leading members of IL-17 signaling pathway. Taken together, our findings suggest that Twist1 plays a role on IL-17 signaling in HER2+ BC.


Influence of the Addition of Extruded Endogenous Tartary Buckwheat Starch on Processing and Quality of Gluten-Free Noodles.
Extruded starch could be used as a thickener for food processing due to its pre-gel properties. This study aimed to explore the influence of the addition of extruded endogenous Tartary buckwheat starch (ES) on the process and quality of gluten-free noodles. ES was mixed with Tartary buckwheat flour in different proportions (10-40%) to prepare the blended flour and noodles. When the content of ES was increased, the swelling power of blended flour at 90 °C had no significant changes, and the decrease in peak viscosity of blended flour was reduced. This indicated that the high-content ES could afford better thermal stability for blended flour and inhibit the swelling ability. The higher level of ES was beneficial to the formation and stabilization of dough, and the improvement of noodle tensile strength. Furthermore, there was no difference in cooking loss between noodles with 30% and 40% ES addition. The microstructure and water distribution of the noodles prepared by blended flour indicated that the gel-entrapped structure organized by the higher content ES could be closely related to the above results. In conclusion, higher ES could contribute to improving the processing properties and quality of noodles.


Clock-hour laminar displacement and age in primary open-angle glaucoma and normal tension glaucoma.
To find out the relationship between laminar displacement and age between patients with primary open-angle glaucoma and normal tension glaucoma. Retrospective study conducted at a tertiary university hospital. Twenty-six eyes of 26 primary open-angle glaucoma patients and 52 eyes of 52 normal tension glaucoma patients. Patients were scanned with a Stratus optical coherence tomography apparatus to measure the retinal nerve fibre layer thickness and to visualize the cross-sectional laminar displacement of 12 clock-hour segments, 30 degrees each. Depth1 was defined as the longest distance between the retinal pigment epithelium and the anterior laminar cribrosa surface, which represents the amount of laminar displacement. Partial correlation coefficients adjusted by mean deviation and intraocular pressure between (i) retinal nerve fibre layer thickness and age, and (ii) Depth1 and age. In the primary open-angle glaucoma group, strong negative correlations (approximately -0.343 ≈ -0.738) were found between Depth1 and age. Eight of 12 clock-hour segments' correlations were significant after Bonferroni correction (α = 0.0021; 24 comparisons). However, no significant correlations were found between Depth1 and age in the normal tension glaucoma group. When the correlation coefficients were compared between the two groups, eight clock-hour segments showed significant differences after Bonferroni correction. The significantly different correlation between laminar displacement and age between primary open-angle glaucoma and normal tension glaucoma patients may suggest a different role of the lamina cribrosa to the disease.


Chiral solvating agents for cyanohydrins and carboxylic acids.
We have shown that a structure as simple as an ion pair of (R)- or (S)-mandelate and dimethylamminopyridinium ions possesses structural features that are sufficient for NMR enantiodiscrimination of cyanohydrins. Moreover, (1)H NMR data of cyanohydrins of known configuration obtained in the presence of the mandelate-dimethylaminopyridinium ion pair point to the existence of a correlation between chemical shifts and absolute configuration of cyanohydrins. Mandelate-DMAPH(+) ion pair and mandelonitrile form a 1:1 complex with an association constant of 338 M(-1) (DeltaG(0), -3.4 kcal/mol) for the (R)-mandelonitrile/(R)-mandelate-DMAPH(+) and 139 M(-1) (DeltaG(0), -2.9 kcal/mol) for the (R)-mandelonitrile/(S)-mandelate-DMAPH(+) complex. To understand the origin of enantiodiscrimination, the geometry optimization and energy minimization of the models of ternary complexes of (S)-mandelonitrile/(R)-mandelate/DMAPH(+) and (S)-mandelonitrile/(S)-mandelate/DMAPH(+) complexes was performed using DFT methodology (B3LYP) with the 6-31+G(d) basis set in Gaussian 3.0. Further, analysis of optimized molecular model obtained from theoretical studies suggested that (i) DMAP may be replaced with other amines, (ii) the hydroxyl group of mandelic acid is not necessary for stabilization of ternary complex and may be replaced with other groups such as methyl, (iii) the ion pair should form a stable ternary complex with any hydrogen-bond donor, provided its OH bond is sufficiently polarized, and (iv) alpha-H of racemic mandelic acid should also get resolved with optically pure mandelonitrile. These inferences were experimentally verified, which not only validated the proposed model but also led to development of a new chiral solvating agent for determination of ee of carboxylic acids and absolute configuration of aryl but not alkyl carboxylic acids.


[Study of the mechanism of supporting cells repairing the organ of Corti in terms of cell kinetics--nuclear DNA synthesis of supporting cell of the organ of Corti in the cochlea damaged by nitromin administration].
3H-thymidine autoradiography in vivo was carried out on normal mature mice, whose organs of Corti were securely damaged by Nitrogen Mustard-N-oxide (nitromin). The cell kinetics of supporting cells of the organ of Corti were examined and relation between these changes and mechanisms of supporting cells to maintain the organ of Corti was discussed in this paper. A few grains in various kinds of supporting cells in S stage, which were not found in control mice without nitromin injection, were discovered. Those were detected in Hensen's cell, Deiters' cell, Claudius' cells, and inner phalangeal cell. Mitosis was seen in one Claudius' cell in addition. Therefore some supporting cells seem to be able to synthesize DNA and proliferate in the acutely damaged organ of Corti even in mature cochlea and those in G0 stage begin to go around cell cycle as occasion demands. Although labelled supporting cells decreased over time, the portion of supporting cell is likely to change in proportion as the extension of damage in the organ of Corti. If this new dynamic change of cell kinetics in the acutely damaged organ of Corti means the reaction to repair the organ of Corti, the supporting cells seem to have a reasonable role to maintain the organ of Corti including repairment of reticular lamina.


Versatile Nanoemulsion Assembly Approach to Synthesize Functional Mesoporous Carbon Nanospheres with Tunable Pore Sizes and Architectures.
Functional mesoporous carbons have attracted significant scientific and technological interest owning to their fascinating and excellent properties. However, controlled synthesis of functional mesoporous carbons with large tunable pore sizes, small particle size, well-designed functionalities, and uniform morphology is still a great challenge. Herein, we report a versatile nanoemulsion assembly approach to prepare N-doped mesoporous carbon nanospheres with high uniformity and large tunable pore sizes (5-37 nm). We show that the organic molecules (e.g., 1,3,5-trimethylbenzene, TMB) not only play an important role in the evolution of pore sizes but also significantly affect the interfacial interaction between soft templates and carbon precursors. As a result, a well-defined Pluronic F127/TMB/dopamine nanoemulsion can be facilely obtained in the ethanol/water system, which directs the polymerization of dopamine into highly uniform polymer nanospheres and their derived N-doped carbon nanospheres with diversely novel structures such as smooth, golf ball, multichambered, and dendritic nanospheres. The resultant uniform dendritic mesoporous carbon nanospheres show an ultralarge pore size (∼37 nm), small particle size (∼128 nm), high surface area (∼635 m2 g-1), and abundant N content (∼6.8 wt %), which deliver high current density and excellent durability toward oxygen reduction reaction in alkaline solution.


HIV reverse transcriptase inhibiting antibodies detected by a new technique: relation to p24 and gp41 antibodies, HIV antigenemia and clinical variables.
A new assay for HIV reverse transcriptase activity inhibiting antibodies (RTI-ab) was used for the analysis of a large collection of sera sampled before and after confirmation of HIV infection. In this assay HIV-RT was preincubated with diluted serum, after which residual RT activity was determined by a technique using a template coupled to macrobeads and 125I-lodo-deoxyuridine-triphosphate as the tracer-substrate. Of the 936 sera analysed, 818 were found positive for RTI-ab, and 824 were positive in Western blot (Wb). The prevalence of RTI-ab compared to Wb was therefore 99.3%. The corresponding figure for 930 sera analysed for envelope-ab, i.e., gp41-ab, was 823 positive, and of these 930 sera 815 were Wb positive, giving a comparative prevalence of 101%. In contrast, only 678 samples of 993 analyzed for core ab, i.e., p24, were positive, giving a prevalence of 77.0% as 880 of these samples were Wb positive. Thus, RTI-ab was as prevalent as gp41-ab, and although the analyses of RTI-ab amounts in different stages showed decreasing levels in stage IV compared to stages II or III, all of the sera except 1 were found positive in stages III and IV. Further, it was found that both the few RTI-ab negative samples in stage II and the few RTI-ab positive samples among Wb negative sera were sampled in connection with seroconversion. The specificity of the RTI-ab assay was 100% in a test of 200 serum samples from HIV negative blood donors. It was concluded that RTI-ab analyses can be made highly sensitive and specific and useful for studies of HIV infection.


Determination of microbial diversity in meju, fermented cooked soya beans, using nested PCR-denaturing gradient gel electrophoresis.
To identify the microbiota in meju, fermented cooked soya beans, that may directly affect the microbial communities of Korean fermented soya bean foods. Using conventional bacterial 16S rDNA, bacilli-specific 16S rDNA or fungi 18S rDNA-specific primers, PCR products were amplified through a series of PCRs using the DNA extracted from ten meju samples. The amplicons were analysed using denaturing gradient gel electrophoresis (DGGE), which showed that Enterococcus durans was commonly detected in nine of ten meju samples. Bacillus subtilis was shown to be the major strain of bacilli in the samples tested. Based on the DGGE analysis of fungi in meju, we determined that Absidia corymbifera, Aspergillus sp. and Candida rugosa were the main fungi in the tested samples. A variety of bacterial and fungal micro-organisms were identified in meju samples, in addition to the micro-organisms already known to be present. This is the first report showing the differences and similarities in the populations of micro-organisms in meju samples using nested PCR-DGGE, a culture-independent method. The results may be applicable to the development of improved meju, in which the indigenous micro-organisms required for fermentation can be standardized.


Transverse myelitis after measles and rubella vaccination.
The incidence of severe neurological complications associated with measles or rubella vaccination is low. A 9-year-old girl developed urinary retention and lower limb paralysis 16 days after measles and rubella vaccination. Her illness was diagnosed as transverse myelitis. Clinical, laboratory and magnetic resonance image findings were consistent with her diagnosis. She was treated with steroids and discharged with only mild lower limb weakness.


Activation of axonal receptors by GABA spillover increases somatic firing.
Axons can be depolarized by ionotropic receptors and transmit subthreshold depolarizations to the soma by passive electrical spread. This raises the possibility that axons and axonal receptors can participate in integration and firing in neurons. Previously, we have shown that exogenous GABA depolarizes cerebellar granule cell axons through local activation of GABA(A) receptors (GABA(A)Rs) and the soma through electrotonic spread of the axonal potential resulting in increased firing. We show here that excitability of granule cells is also increased by release of endogenous GABA from molecular layer interneurons (MLIs) and spillover activation of parallel fiber GABA(A)Rs in mice and rats. Changes in granule cell excitability were assessed by excitability testing after activation of MLIs with channelrhodopsin or electrical stimulation in the molecular layer. In granule cells lacking an axon, excitability was not changed, suggesting that axonal receptors are required. To determine the distance over which subthreshold potentials may spread, we estimated the effective axonal electrical length constant (520 μm) by excitability testing and focal uncaging of RuBi-GABA on the axon at varying distances from the soma. These data suggest that GABA(A)R-mediated axonal potentials can participate in integration and firing of cerebellar granule cells.


Symmetrization of cationic hydrogen bridges of protonated sponges induced by solvent and counteranion interactions as revealed by NMR spectroscopy.
The properties of the intramolecular hydrogen bonds of doubly (15)N-labeled protonated sponges of the 1,8-bis(dimethylamino)naphthalene (DMANH(+)) type have been studied as a function of the solvent, counteranion, and temperature using low-temperature NMR spectroscopy. Information about the hydrogen-bond symmetries was obtained by the analysis of the chemical shifts delta(H) and delta(N) and the scalar coupling constants J(N,N), J(N,H), J(H,N) of the (15)NH(15)N hydrogen bonds. Whereas the individual couplings J(N,H) and J(H,N) were averaged by a fast intramolecular proton tautomerism between two forms, it is shown that the sum |J(N,H)+J(H,N)| generally represents a measure of the hydrogen-bond strength in a similar way to delta(H) and J(N,N). The NMR spectroscopic parameters of DMANH(+) and of 4-nitro-DMANH(+) are independent of the anion in the case of CD(3)CN, which indicates ion-pair dissociation in this solvent. By contrast, studies using CD(2)Cl(2), [D(8)]toluene as well as the freon mixture CDF(3)/CDF(2)Cl, which is liquid down to 100 K, revealed an influence of temperature and of the counteranions. Whereas a small counteranion such as trifluoroacetate perturbed the hydrogen bond, the large noncoordinating anion tetrakis[3,5-bis(trifluoromethyl)phenyl]borate B[{C(6)H(3)(CF(3))(2)}(4)](-) (BARF(-)), which exhibits a delocalized charge, made the hydrogen bond more symmetric. Lowering the temperature led to a similar symmetrization, an effect that is discussed in terms of solvent ordering at low temperature and differential solvent order/disorder at high temperatures. By contrast, toluene molecules that are ordered around the cation led to typical high-field shifts of the hydrogen-bonded proton as well as of those bound to carbon, an effect that is absent in the case of neutral NHN chelates.


Outcomes of pediatric mixed phenotype acute leukemia treated with lymphoid directed therapy: Analysis of an institutional series from India.
There is limited data regarding pediatric mixed phenotype acute leukemia (MPAL) and there is no global consensus on its management yet. In this retrospective study, we analyzed the outcomes of children diagnosed with MPAL at our institute. This study included children ≤ 14 years with MPAL who presented to a tertiary cancer center in India from January 1st 2009 to December 31st 2015. Over a seven-year period, 1390 patients with leukemia presented to our institute of which 22 patients (1.5%) had MPAL. Sixteen patients (72.7%) had B/myeloid leukemia, while 4 (18.1%) and 2 (9%) patients had T/myeloid and B/T leukemia respectively. Twenty-one patients were treated with a modified BFM ALL 95 protocol. 76.1% (n = 16) of patients had a good prednisolone response (GPR) on day 8 and end-of-induction (EOI) marrow was in remission in 90.5% (n = 19). A poor prednisolone response (PPR) on day 8 correlated with an inferior relapse-free survival (25% vs 79.5%, P=.025). The 4-year event-free survival (EFS) and overall survival (OS) for the entire group was 60.8% and 64.9% respectively while the EFS for patients who had a GPR and remission at the EOI (n = 15) was 80% as compared to 16.7% in patients with PPR or induction failure. Lymphoid directed chemotherapy is seen to have good survival outcomes in pediatric MPAL. However, a PPR on day 8 or a positive EOI marrow may be an indication for more aggressive treatment.


Impacts of fludioxonil resistance on global gene expression in the necrotrophic fungal plant pathogen Sclerotinia sclerotiorum.
The fungicide fludioxonil over-stimulates the fungal response to osmotic stress, leading to over-accumulation of glycerol and hyphal swelling and bursting. Fludioxonil-resistant fungal strains that are null-mutants for osmotic stress response genes are easily generated through continual sub-culturing on sub-lethal fungicide doses. Using this approach combined with RNA sequencing, we aimed to characterise the effects of mutations in osmotic stress response genes on the transcriptional profile of the important agricultural pathogen Sclerotinia sclerotiorum under standard laboratory conditions. Our objective was to understand the impact of disruption of the osmotic stress response on the global transcriptional regulatory network in an important agricultural pathogen. We generated two fludioxonil-resistant S. sclerotiorum strains, which exhibited growth defects and hypersensitivity to osmotic stressors. Both had missense mutations in the homologue of the Neurospora crassa osmosensing two component histidine kinase gene OS1, and one had a disruptive in-frame deletion in a non-associated gene. RNA sequencing showed that both strains together differentially expressed 269 genes relative to the parent during growth in liquid broth. Of these, 185 (69%) were differentially expressed in both strains in the same direction, indicating similar effects of the different point mutations in OS1 on the transcriptome. Among these genes were numerous transmembrane transporters and secondary metabolite biosynthetic genes. Our study is an initial investigation into the kinds of processes regulated through the osmotic stress pathway in S. sclerotiorum. It highlights a possible link between secondary metabolism and osmotic stress signalling, which could be followed up in future studies.


Analysis of cognitive and motor functioning during pubertal development: a new approach.
We investigated cognitive-motor abilities in 303 (156 female) school children from Zagreb, Croatia, in the age span 10 to 14 years using a newly developed chronometrical reactionmeter system (CRD). The following tests were applied: CRD-311 (simple visual discrimination of signal location), CRD-324 (short-term memory actualisation), CRD-21 (simple convergent visual orientation), and CRD-11 (arithmetically conceptualised/operationalised convergent thinking). In both gender a statistically significant age related improvement of the performance for time related parameters (minimum time of test item solving (MT), total ballast (TB), and total time of test solving (TT) was observed. In contrast, the number of errors (NE), which was the only non-time related parameter tested, did not significantly change with age. Significant differences between boys and girls were observed for the time related parameters TB and MT. TB was significantly lower in girls, whereas boys tended to be faster in MT measurements. In TT as a composed measure of the mentioned parameters, no major differences were observed. We conclude that the CRD system is a new useful tool for investigating the complexity of cognitive-motor abilities in children. Our cross-sectional study demonstrated that the time-related parameters were significantly affected by age and gender during puberty.


Overactive bladder.
Overactive bladder (OAB) is a common condition (prevalence 14%-18% of Canadians) and has a significant negative impact on quality of life. OAB may be idiopathic or may occur with other common conditions such as bladder outlet obstruction, neurological disease, or stress incontinence. Primary care physicians may safely diagnose this condition by history and physical exam with a minimum of widely available lab tests. Management with behavioral therapies and pharmacotherapy is generally quite successful and warranted. Multiple anticholinergic medications are available and have been shown to be effective. Subtle differences in structure and mechanism of these agents may yield improved therapeutic benefit or tolerability and thus it is reasonable to try more than one drug to achieve the optimal results. For patients that fail behavioral and initial pharmacotherapy or when other complicating conditions are identified, referral to a specialist is indicated; however, the majority of patients with OAB do not require cystoscopy or urodynamics. Successful treatments for OAB do exist and it is worth screening for these disabling complaints at the primary care level.


Modulation of cardiopulmonary depressor reflex in nucleus ambiguus by electroacupuncture: roles of opioids and γ-aminobutyric acid.
Stimulation of cardiopulmonary receptors with phenylbiguanide (PBG) elicits depressor cardiovascular reflex responses, including decreases in blood pressure and heart rate mediated in part by the brain stem parasympathetic cardiac neurons in the nucleus ambiguus (NAmb). The present study examined NAmb neurotransmitter mechanisms underlying the influence of electroacupuncture (EA) on the PBG-induced hypotension and bradycardia. We hypothesized that somatic stimulation during EA modulates PBG responses through opioid and γ-aminobutyric acid (GABA) modulation in the NAmb. Anesthetized and ventilated cats were studied during repeated stimulation with PBG or cardiac vagal afferents while low-frequency EA (2 Hz) was applied at P5-6 acupoints overlying the median nerve for 30 min and NAmb neuronal activity, heart rate, and blood pressure were recorded. Microinjection of kainic acid into the NAmb attenuated the PBG-induced bradycardia from -60 ± 11 to -36 ± 11 beats/min. Likewise, EA reduced the PBG-induced depressor and bradycardia reflex by 52 and 61%, respectively. Cardiac vagal afferent evoked preganglionic cellular activity in the NAmb was reduced by EA for about 60 min. Blockade of opioid or GABA(A) receptors using naloxone and gabazine reversed the EA-related modulation of the evoked cardiac vagal activity by 73 and 53%, respectively. Similarly, naloxone and gabazine reversed EA modulation of the negative chronotropic responses from -11 ± 5 to -23 ± 6 and -13 ± 4 to -24 ± 3 beats/min, respectively. Thus EA at P5-6 decreases PBG evoked hypotension and bradycardia as well as the NAmb PBG-sensitive preganglionic cardiac vagal outflow through opioid and GABA neurotransmitter systems.


Health-Related Quality-of-Life Outcomes after Transoral Robotic Surgery for T1 and T2 Supraglottic Laryngeal Carcinoma Compared to the Transcervical Open Supraglottic Approach.
This prospective controlled study evaluated and compared health-related quality of life (HRQOL), after transoral robotic supraglottic laryngectomy (TORSGL) versus transcervical open supraglottic laryngectomy (TCOSGL) in patients with T1 and T2 supraglottic laryngeal carcinoma (SLC). The TORSGL group comprised 14 patients, and the TCOSGL group comprised 13 patients. All 27 patients completed the European Organization for Research and Treatment of Cancer 30-item core quality-of-life questionnaire version 3.0 (EORTC QLQ-C30), the European Organization for Research and Treatment of Cancer head-and-neck cancer-specific module (EORTC QLQ-H&N35), before treatment and during the early and the late postoperative periods. The present prospective study demonstrated the near-term postoperative HRQOL of patients with T1 or T2 SLC treated with TORSGL (Group A) or TCOSGL (Group B). On comparison of EORTC QLQ-C30 data for the two groups in the early postoperative period, all functional subscale scores and global health status scores were statistically significantly lower (all p < 0.05) in Group A than in Group B and in the late postoperative period, other than the cognitive function score (p = 0.450), all functional subscale scores and global health status scores were statistically significantly lower (all p < 0.05) in Group A than in Group B. On comparison of the EORTC QLQ-H&N35 data for the two groups in the early postoperative period, except for the teeth problems scale score (p = 0.061), all symptom scale scores were statistically significantly lower in Group A than in Group B (all p < 0.05) and in the late postoperative period, the speech, social eating, social contact, and coughing scale scores were statistically significantly lower (p = 0,0215, p = 0.021, p = 0.01, p = 0.011, respectively) in Group A than in Group B. HRQOL parameters recovered in the late postoperative period in both groups; recovery was better in Group A. This study suggested that TORSGL may provide patients with a better HRQOL than those TCOSGL, especially in the early period, but also in the late period.


The triple clinicopathologic features to seminal vesicle-sparing radical prostatectomy.
With the widespread early detection programs for prostate cancer, there has been a downward stage migration and a marked decrease in the percentage of men with seminal vesicle invasion (SVI) compared with previous data. We evaluated clinicopathologic findings that are associated with SVI to select patients for potential seminal vesicle-sparing surgery. We reviewed our radical prostatectomy database from 1997 to 2006 to evaluate the incidence and clinical correlates of SVI. Variables analyzed included serum prostate-specific antigen (PSA) level, clinical stage, percentage of positive cores with cancer, Gleason score on biopsy, age, prostate weight, and urethral and vesical surgical margins. Statistical analysis included univariate and multivariate logistic regressions. Of 267 patients, 32 (12%) had SVI. Preoperative PSA level, biopsy Gleason score, and percentage of positive cores were highly predictive of SVI on multivariate analysis. SVI was present in only 1/98 patients (1.02 %) with biopsy Gleason score ≤6, 0/23 patients (0%) with serum PSA level <4 ng/mL, and only 1 patient with ≤12.8% of positive cores on biopsy. In all cases of distal SVI, there was proximal involvement. Serum PSA level, Gleason score, and percentage of positive cores on biopsy are statistically significant predictors of SVI on multivariate analysis. Seminal vesiculectomy does not benefit almost 99% of patients with biopsy Gleason score ≤6, PSA level <4 ng/mL, and with <12% cores with cancer. In cases of seminal vesicle-sparing surgery, frozen section of the proximal portion may be of adjunct usefulness for the triple.


Intereleukin-10 promoter polymorphism in mild cognitive impairment and in its clinical evolution.
Specific proinflammatory alleles are associated with higher risk of Alzheimer disease (AD) in different onset age. The homozygosis for the A allele of -1082 polymorphism (G/A) of interleukin-10 (IL-10) promotes a higher risk of AD and reduced IL-10 generation in peripheral cells after amyloid stimulation. In this paper we analysed genotype and allele frequencies of this polymorphism in 138 subjects with mild cognitive impairment (MCI) diagnosed, respectively, as amnestic (a-MCI) and multiple impaired cognitive domains (mcd-MCI). The genotype frequencies were similar in a-MCI and AD subjects, whereas in mcd-MCI comparable to controls (AA genotype: 50% in a-MCI, 49.2% in AD, 28.7% in mcd-MCI and 31.8% in controls). Consequently, both allele and genotype distributions were significantly different between a-MCI and mcd-MCI (allele: P = .02, genotype: P < .05). These results support the theory that polymorphisms of cytokine genes can affect neurodegeneration and its clinical progression. IL-10 may partly explain the conversion of a-MCI to AD or be a genetic marker of susceptibility.