Salmonella-based platform for efficient delivery of functional binding proteins to the cytosol.
Protein-based affinity reagents (like antibodies or alternative binding scaffolds) offer wide-ranging applications for basic research and therapeutic approaches. However, whereas small chemical molecules efficiently reach intracellular targets, the delivery of macromolecules into the cytosol of cells remains a major challenge; thus cytosolic applications of protein-based reagents are rather limited. Some pathogenic bacteria have evolved a conserved type III secretion system (T3SS) which allows the delivery of effector proteins into eukaryotic cells. Here, we enhance the T3SS of an avirulent strain of Salmonella typhimurium to reproducibly deliver multiple classes of recombinant proteins into eukaryotic cells. The efficacy of the system is probed with both DARPins and monobodies to functionally inhibit the paradigmatic and largely undruggable RAS signaling pathway. Thus, we develop a bacterial secretion system for potent cytosolic delivery of therapeutic macromolecules.


Functionalized ZnO/ZnO2 n-N straddling heterostructure achieved by oxygen plasma bombardment for highly selective methane sensing.
Metal oxide semiconductors have been extensively used as reducing gas sensors with major limitations regarding selectivity and operating temperature which is relatively high for most of the cases making the device unusable in some critical situations. Higher operating temperature is also associated with the higher power consumption, which goes against the miniaturization of the device. In order to resolve these problems, here we introduced a ZnO/ZnO2 straddling 'n-N' isotype heterostructure as a highly selective and sensitive methane sensor at moderately low operating temperature. ZnO-Zn(OH)2 precursor films were treated in oxygen plasma in a pulsed DC magnetron sputtering system. Morphological analyses by field emission scanning electron microscopy showed flake like growth of the grains with high surface roughness, whereas X-ray diffraction (XRD) showed polycrystalline nature of the films. Polycrystalline ZnO2 peaks were observed in the XRD pattern in addition to the existing ZnO, which indicates modification of the precursor to oxygen rich heterostructure of ZnO/ZnO2. This was further supported by the shifting of the O1s peak in the X-ray photoelectron spectroscopic analysis. Plasma treated ZnO/ZnO2 heterostructured films were found to show high selectivity towards methane (with respect to H2S and CO) and sensitivity (∼96%) at a comparatively low operating temperature.


Visualizing copper assisted graphene growth in nanoscale.
Control synthesis of high quality large-area graphene on transition metals (TMs) by chemical vapor deposition (CVD) is the most fascinating approach for practical device applications. Interaction of carbon atoms and TMs is quite critical to obtain graphene with precise layer number, crystal size and structure. Here, we reveal a solid phase reaction process to achieve Cu assisted graphene growth in nanoscale by in-situ transmission electron microscope (TEM). Significant structural transformation of amorphous carbon nanofiber (CNF) coated with Cu is observed with an applied potential in a two probe system. The coated Cu particle recrystallize and agglomerate toward the cathode with applied potential due to joule heating and large thermal gradient. Consequently, the amorphous carbon start crystallizing and forming sp(2) hybridized carbon to form graphene sheet from the tip of Cu surface. We observed structural deformation and breaking of the graphene nanoribbon with a higher applied potential, attributing to saturated current flow and induced Joule heating. The observed graphene formation in nanoscale by the in-situ TEM process can be significant to understand carbon atoms and Cu interaction.


What Drives Internet Entrepreneurial Intention to Use Technology Products? An Investigation of Technology Product Imagination Disposition, Social Support, and Motivation.
Technological products such as computer, communication, and consumer electronic products, apps, smart wearables, and streaming services have become inseparable from people's lives. In technological fields of practice, imagination, creativity, innovation, and entrepreneurship may influence one another. A vivid imagination can generate creativity and trigger the entrepreneurial intention to "bring new things to the market." This study aims to understand the formation of internet entrepreneurial intention to use technology products. Drawing on social cognitive theory, this study explores and empirically tests how technology product imagination disposition and social support impact internet entrepreneurial intention to use technology products. Drawing from self-determination theory, this study proposes and examines the mediating role of intrinsic (challenge and enjoyment) and extrinsic motivation (compensation and outward motivation) in the relationship between technology product imagination disposition and internet entrepreneurial intention, as well as the relationship between social support and internet entrepreneurial intention. We conducted a survey of 568 adults in Taiwan and used partial least squares to test our hypotheses. The results show the following: (1) Technology product imagination disposition is positively associated with challenge, enjoyment, compensation, and outward motivation. (2) Social support is positively associated with challenge, enjoyment, compensation, and outward motivation. (3) Challenge, enjoyment, and outward motivation are positively associated with internet entrepreneurial intention to use technology products. (4) Technology product imagination disposition intensifies internet entrepreneurial intention to use technology products by strengthening challenge, enjoyment, and outward motivation. Social support intensifies internet entrepreneurial intention by increasing challenge, enjoyment, and outward motivation. The partial mediation model represents a significant improvement in the total effect over the direct effect. We discuss the implications of these results for research and internet entrepreneurship practices.


High-Intensity Statin Therapy Is Associated With Improved Survival in Patients With Peripheral Artery Disease.
The relative benefit of higher statin dosing in patients with peripheral artery disease has not been reported previously. We compared the effectiveness of low- or moderate-intensity (LMI) versus high-intensity (HI) statin dose on clinical outcomes in patients with peripheral artery disease. We reviewed patients with symptomatic peripheral artery disease who underwent peripheral angiography and/or endovascular intervention from 2006 to 2013 who were not taking other lipid-lowering medications. HI statin use was defined as atorvastatin 40-80 mg or rosuvastatin 20-40 mg. Baseline demographics, procedural data, and outcomes were retrospectively analyzed. Among 909 patients, 629 (69%) were prescribed statins, and 124 (13.6%) were treated with HI statin therapy. Mean low-density lipoprotein level was similar in patients on LMI versus HI (80±30 versus 87±44 mg/dL, P=0.14). Demographics including age (68±12 versus 67±10 years, P=0.25), smoking history (76% versus 80%, P=0.42), diabetes mellitus (54% versus 48%, P=0.17), and hypertension (88% versus 89%, P=0.78) were similar between groups (LMI versus HI). There was a higher prevalence of coronary artery disease (56% versus 75%, P=0.0001) among patients on HI statin (versus LMI). After propensity weighting, HI statin therapy was associated with improved survival (hazard ratio for mortality: 0.52; 95% confidence interval, 0.33-0.81; P=0.004) and decreased major adverse cardiovascular events (hazard ratio: 0.58; 95% confidence interval 0.37-0.92, P=0.02). In patients with peripheral artery disease who were referred for peripheral angiography or endovascular intervention, HI statin therapy was associated with improved survival and fewer major adverse cardiovascular events compared with LMI statin therapy.


Analysis of Pelvis-Thorax Coordination Patterns of Professional and Amateur Golfers during Golf Swing.
The aim of this research was to quantify the coordination pattern between thorax and pelvis during a golf swing. The coordination patterns were calculated using vector coding technique, which had been applied to quantify the coordination changes in coupling angle (γ) between two different segments. For this, fifteen professional and fifteen amateur golfers who had no significant history of musculoskeletal injuries. There was no significant difference in coordination patterns between the two groups for rotation motion during backswing (p = 0.333). On the other hand, during the downswing phase, there were significant differences between professional and amateur groups in all motions (flexion/extension: professional [γ] = 187.8°, amateur [γ] = 167.4°; side bending: professional [γ] = 288.4°, amateur [γ] = 245.7°; rotation: professional [γ] = 232.0°, amateur [γ] = 229.5°). These results are expected to be a discriminating measure to assess complex coordination of golfers' trunk movements and preliminary study for interesting comparison by golf skilled levels.


High dose calcitriol may reduce thrombosis in cancer patients.
The incidence of venous and arterial thrombosis in a placebo-controlled randomised trial of DN-101 (high dose calcitriol) with docetaxel versus docetaxel was compared. Of the 13 thrombotic events observed in the 250 patients enroled in this study, two occurred in DN-101 and 11 in placebo-treated patients (P = 0.01). This difference remained significant after adjustment for baseline history of thrombosis, atrial fibrillation and use of anti-thrombotic agents. In vitro and vitamin D receptor (VDR) knockout mouse studies predict that nanomolar concentrations of calcitriol may act as an antithrombotic agent. We report the first clinical observation that supports this hypothesis in humans.


['Vegetable wasps and plant worms' and G-protein-coupled receptors].
Following the discovery of an immunosuppressive substance isolated from one of the entomopathogenic fungi 'vegetable wasps and plant worms', FTY720, whose immunosuppressive action is more potent than that of cyclosporin, was developed. In contrast to classical immunosuppressants such as cyclosporin, FTY720 does not affect the growth, maturation and activation of lymphocytes, but causes a reduction in the number of circulating lymphocytes as a result of enhanced homing of lymphocytes to the secondary lymphoid organs. It has recently been reported that FTY720 is phosphorylated in vivo by sphingosine kinase, and the phosphorylated FTY720 is a ligand of the receptors of sphingosine 1-phosphate (S1P), which are G-protein-coupled receptors. S1P has been recognized most prominently as a platelet-derived lipid mediator which regulates several functions of endothelial and smooth muscle cells. In addition, recent studies have suggested the crucial roles for S1P in immunological, and inflammatory modulation.


Subthalamic nucleus lesions enhance the psychomotor-activating, incentive motivational, and neurobiological effects of cocaine.
The subthalamic nucleus (STN) is traditionally thought to be involved in motor control, and dysfunction of the STN is thought to contribute to movement disorders. Here, we show that the STN also plays an important role in motivational processes and the response to drugs of abuse. Specifically, bilateral STN lesions produced a dose-dependent increase in the psychomotor-activating effects of cocaine, the rate at which animals acquired cocaine self-administration, and the motivation for cocaine assessed using a progressive ratio schedule. Furthermore, bilateral STN lesions enhanced the ability of cocaine to induce gene expression in the nucleus accumbens and caudate-putamen, two structures known to be involved in mediating the psychomotor-activating and incentive motivational effects of drugs of abuse. These findings suggest that engagement of the STN serves to dampen the psychomotor-activating and incentive motivational effects of drugs of abuse. Thus, the STN may serve as a novel target for therapeutic interventions aimed at treating drug dependence.


Pediatric impetigo: an expert panel opinion about its main controversies.
Bacterial impetigo is one of the most common skin infection in childhood. Uncertainty exists about its management. This article offers practical suggestions, given the existing evidence and experts' opinions, for correctly managing pediatric impetigo in both hospital and ambulatory settings. Italian physicians with an expertise on pediatric impetigo appointed a working group. A preliminary literature search using Pubmed/MEDLINE and Cochrane Library databases has been performed. The most common controversial issues about pediatric impetigo have been identified and then discussed from multidisciplinary perspectives, according to the 'structured controversy' methodology, a technique discovered and designed to get engaged in a controversy and then guide participants to seek consensus. The expert panels identified 10 main controversies about pediatric impetigo. All of them have been discussed from dermatological, pediatric, pharmacological and microbiological points of view reaching consensus. Each controversy has been revised thus giving practical issues for an easy use in clinical practice. Based on clinical experts' opinion, local epidemiology and literature review this article offers practical suggestions for the management of pediatric impetigo trying to reduce uncertainty in this setting of care.


Suicide attempt using potassium tablets for congenital chloride diarrhea: A case report.
Congenital chloride diarrhea (CCD) is a rare inherited disorder of intestinal electrolyte transport that results in a large wastage of electrolytes and water. Advances in substitution therapy using sodium chloride (NaCl) and potassium chloride (KCl) have dramatically improved survival for patients with CCD. Slow-release KCl is widely prescribed as a potassium supplement; however, it has also occasionally been used in suicide attempts, as potassium poisoning can generate life-threatening hyperkalemia. A 26-year-old female presented to the emergency department (ED) with self-poisoning, having taken 30 tablets of slow-release KCl (total: 240 mmol potassium) following an auditory hallucination. The patient had been undergoing substitution therapy with NaCl and KCl for CCD and been followed up in the pediatric department. One month prior, she developed insomnia and anxiety and had consulted a psychiatrist. At the ED, although her general condition was good, she appeared agitated. Her serum potassium level was 7.0 mmol/L, indicating hyperkalemia, and electrocardiographic changes showed tenting of the T-waves. She responded to the administration of calcium gluconate, sodium bicarbonate, and insulin with glucose, and the serum potassium level improved. Finally, she was diagnosed with schizophrenia. In CCD management, physicians should pay careful attention to patients' extraintestinal issues, including psychological disorders that may emerge in adulthood.


Histidine-rich glycoprotein plus zinc to neutralize heparin.
Histidine-rich glycoprotein (HRG) binds heparin and neutralizes its anticoagulant effect. Because zinc greatly enhances this interaction, it is possible that the combination of HRG and zinc could be used as an antidote for heparin. We have determined the plasma concentrations of HRG and zinc necessary to neutralize clinically relevant concentrations of heparin. Using a thrombin-time assay, we found that HRG plus zinc can neutralize 0.2 to 4.5 units/mL heparin, although the maximal effect requires an HRG plasma concentration about six times normal and a zinc concentration about 10 times normal. In a system using purified proteins and a thrombin-specific chromogenic substrate, we found that both the concentration of HRG and the molar ratio of HRG to zinc are important in determining the potency of the anti-heparin activity. Whether HRG and zinc can be administered as an antidote for heparin will depend on whether the requisite doses can be achieved without toxicity.


Place of work, communities and municipalities.
The Paris conference recognized that although national social policy and international research were undoubtedly important, a local dimension to research action and planning was essential. The potential for community and municipal action was illustrated in a number of papers which are summarized here. It was also the subject of a publication in the series associated with the European Alcohol Action Plan. The work-place was taken as a significant microcosm of a community, where healthy company policy and an enlightened approach to providing early help and support to individuals with alcohol-related problems is particularly beneficial.


Signaling pathways for tissue factor expression in lipopolysaccharide-stimulated bovine alveolar macrophages.
To investigate receptor-mediated intracellular events in bovine alveolar macrophages (AM) stimulated by bacterial lipopolysaccharide (LPS), using tissue factor (TF) expression as the measurable functional endpoint. Pulmonary AM harvested from 1- to 4-month-old male Holstein calves. Alveolar macrophages, acquired by use of volume-controlled bronchopulmonary lavage, were treated with CD14 monoclonal antibody (20 microg/ml), pertussis toxin (300 ng/ml), or 1 of 3 known protein kinase C (PKC) inhibitors (10 microM chelerythrin, 100 microM H-7, or 50 nM staurosporin), then were stimulated with LPS alone (0.01, 0.10, 1.0, 10.0 microg/ml) or LPS (0.25, 0.5, 1.0 ng/ml) in combination with concentrated bovine serum fraction 2 (500 ng/ml). Tissue factor expression was quantified by use of a colorimetric assay. Changes in intracellular Ca2+ concentration and pH were monitored, using Ca2+- and pH-sensitive fluorescent dyes, with changes in fluorescent intensity after incubation with LPS measured by spectrophotometry. Treatment of AM with a CD14 monoclonal antibody caused profound inhibition of TF expression (P < 0.0001) after stimulation by LPS combined with bovine serum fraction 2. Pertussis toxin had a significant (P < 0.0319) inhibitory effect on TF expression when cells were stimulated by LPS alone. Treatment with all 3 PKC inhibitors caused marked reduction in TF expression of cells stimulated with LPS alone or with phorbol myristate acetate. Stimulation of cells by LPS failed to mobilize intracellular Ca2+ stores or to alter cytosolic pH. LPS combined with serum factors binds to CD14 on the surface of AM, and PKC is an important signaling kinase in the pathway utilized by LPS, resulting in enhanced TF expression; a pertussis toxin-sensitive G protein is involved in the signaling pathway utilized by LPS alone; and mobilization of Ca2+ does not have a role in the signal transduction pathway utilized by LPS nor does LPS affect cytosolic pH of AM.


Maintenance of muscle mass is not dependent on the calcineurin-NFAT pathway.
In this study, the role of the calcineurin pathway in skeletal muscle atrophy and atrophy-reducing interventions was investigated in rat soleus muscles. Because calcineurin has been suggested to be involved in skeletal and cardiac muscle hypertrophy, we hypothesized that blocking calcineurin activity would eliminate beneficial effects of interventions that maintain muscle mass in the face of atrophy-inducing stimuli. Hindlimb suspension and spinal cord transection were used to induce atrophy, and intermittent reloading and exercise were used to reduce atrophy. Cyclosporin (CsA, 25 mg x kg(-1) x day(-1)) was administered to block calcineurin activity. Soleus muscles were studied 14 days after the onset of atrophy. CsA administration did not inhibit the beneficial effects of the two muscle-maintaining interventions, nor did it change muscle mass in control or atrophied muscles, suggesting that calcineurin does not play a role in regulating muscle size during atrophy. However, calcineurin abundance was increased in atrophied soleus muscles, and this was associated with nuclear localization of NFATc1 (a nuclear factor of activated T cells). Therefore, results suggest that calcineurin may be playing opposing roles during skeletal muscle atrophy and under muscle mass-maintaining conditions.


Portal vein computed tomography imaging characteristics and their relationship with bleeding risk in patients with liver cirrhosis undergoing interventional therapy.
This study aimed to analyze the predictive value of multi-slice spiral computed tomography (CT) perfusion imaging for upper gastrointestinal bleeding in patients with cirrhotic portal hypertension. A total of 62 patients with cirrhotic portal hypertension and 28 healthy individuals were included. The results showed that multi-slice spiral CT perfusion imaging had a significant predictive value for upper gastrointestinal bleeding in patients with cirrhotic portal hypertension. The vascular area, number of vascular cross-sections, and gastric coronary vein diameter (GCVD) showed high predictive values, with the vascular area having the best predictive value. To investigate the predictive accuracy of multi-slice spiral CT perfusion imaging for upper gastrointestinal bleeding in patients with cirrhosis and portal hypertension. This study included 62 patients with cirrhotic portal hypertension (disease group) and 28 healthy individuals (control group). The disease group was further divided into two subgroups: Group A (n = 27, bleeding) and group B (n = 35, no bleeding). All patients underwent multi-slice spiral CT perfusion imaging at our hospital, and we compared various parameters such as liver blood flow, vein size, number of blood vessels, and blood vessel area between the two groups. We employed statistical analysis to identify factors associated with upper gastrointestinal bleeding and created a graph comparing the predictive value of different factors for bleeding. We found no difference in hepatic artery (HAP) levels among the three groups (all P > 0.05). The portal vein levels in groups A and B were much lower than in the control group; group A was much lower than group B (all P < 0.05). The HAP perfusion index levels in groups A and B were much higher than in the control group; group A was much higher than group B (all P < 0.05). The portal vein diameter, splenic vein diameter, and GCVD levels in groups A and B were much higher than in the control group; those in group A were much higher than those in group B (all P < 0.05). The number of blood vessels and blood vessel area in groups A and B were much higher than in the control group; those in group A were much higher than those in group B (all P < 0.05). The statistical method showed a strong link between GCVD, number of blood vessels, blood vessel area, and upper gastrointestinal bleeding (odds ratio = 1.275, 1.346, 1.397, P < 0.05). The graph showed that GCVD, number of blood vessels, and blood vessel area could predict bleeding well, with blood vessel area having the best prediction power. That multi-slice spiral CT perfusion imaging can predict upper gastrointestinal bleeding well in patients with cirrhosis and high blood pressure in the portal vein. GCVD, number of blood vessels, and blood vessel area had high prediction power. The blood vessel area had the best prediction power, with an area under the curve of 0.831.


Lipid exchange and transfer on nanoparticle supported lipid bilayers: effect of defects, ionic strength, and size.
Lipid exchange/transfer has been compared for zwitterionic 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2-dimyristoyl-d54-sn-glycero-3-phosphocholine (DMPC) small unilamellar vesicles (SUVs) and for the same lipids on silica (SiO2) nanoparticle supported lipid bilayers (NP-SLBs) as a function of ionic strength, temperature, temperature cycling, and NP size, above the main gel-to-liquid crystal phase transition temperature, Tm, using d- and h-DMPC and DPPC. Increasing ionic strength decreases the exchange kinetics for the SUVs, but more so for the NP-SLBs, due to better packing of the lipids and increased attraction between the lipid and support. When the NP-SLBs (or SUVs) are cycled above and below Tm, the exchange rate increases compared with exchange at the same temperature without cycling, for similar total times, suggesting that defects provide sites for more facile removal and thus exchange of lipids. Defects can occur: (i) at the phase boundaries between coexisting gel and fluid phases at Tm; (ii) in bare regions of exposed SiO2 that form during NP-SLB formation due to mismatched surface areas of lipid and NPs; and (iii) during cycling as the result of changes in area of the lipids at Tm and mismatched thermal expansion coefficient between the lipids and SiO2 support. Exchange rates are faster for NP-SLBs prepared with the nominal amount of lipid required to form a NP-SLB compared with NP-SLBs that have been prepared with excess lipids to minimize SiO2 patches. Nanosystems prepared with equimolar mixtures of NP-SLBs composed of d-DMPC (d(DMPC)-NP-SLB) and SUVs composed of h-DMPC (h(DMPC)-SUV) show that the calorimetric transition of the "donor" h(DMPC)-SUV decreases in intensity without an initial shift in Tm, indicating that the "acceptor" d(DMPC)-NP-SLB can accommodate more lipids, through either further fusion or insertion of lipids into the distal monolayer. Exchange for d/h(DMPC)-NP-SLB is in the order 100 nm SiO2 > 45 nm SiO2 > 5 nm SiO2.


Cystic hemangioma of the spleen in a schizophrenic patient.
We have presented a single case of cavernous hemangioma of the spleen with cystic formation. We believe this is the first reported case associated with schizophrenia. Splenic scan demonstrated only a thin rim of functioning tissue. In the absence of a reliable history, celiac angiography provided the most important preoperative information on this large, barely functioning cystic spleen. A generous incision gave the required exposure for removal. The size of the cyst and the amount of compression and deviation of the splenic artery encouraged us to use a thoracoabdominal incision. We believe this is the 75th case of cystic hemangioma of the spleen reported in the literature.


The dynamic nature of the nuclear envelope: lessons from closed mitosis.
In eukaryotes, chromosomes are encased by a dynamic nuclear envelope. In contrast to metazoans, where the nuclear envelope disassembles during mitosis, many fungi including budding yeast undergo "closed mitosis," where the nuclear envelope remains intact throughout the cell cycle. Consequently, during closed mitosis the nuclear envelope must expand to accommodate chromosome segregation to the two daughter cells. A recent study by Witkin et al. in budding yeast showed that if progression through mitosis is delayed, for example due to checkpoint activation, the nuclear envelope continues to expand despite the block to chromosome segregation. Moreover, this expansion occurs at a specific region of the nuclear envelope- adjacent to the nucleolus- forming an extension referred to as a "flare." These observations raise questions regarding the regulation of nuclear envelope expansion both in budding yeast and in higher eukaryotes, the mechanisms confining mitotic nuclear envelope expansion to a particular region and the possible consequences of failing to regulate nuclear envelope expansion during the cell cycle.


Impact of four (13)C-proline isotope labels on the infrared spectra of ribonuclease T1.
Ribonuclease T1 was biosynthesized, with all four prolines (13)C-labeled in the peptide C[double bond]O bond, using a proline auxotrophic yeast strain of Saccharomyces cerevisiae. The (13)C- and (12)C-proline isotopomers of ribonuclease T1 were investigated by infrared spectroscopy in the thermally unfolded and natively folded state at 80 and 20 degrees C, respectively. In the thermally unfolded state, both proteins established almost indistinguishable spectral features in the secondary structure sensitive amide I region. In contrast, the spectra measured at 20 degrees C revealed substantial qualitative and quantitative differences, though parallel analysis by circular dichroism suggested identical native folds for both isotopomers. Major spectral differences in the infrared spectra were detected at 1626 and 1679 cm(-1), which are diagnostic marker bands for antiparallel beta-sheets in ribonuclease T1 and at 1645 cm(-1), a region that is characteristic for the infrared absorption of irregular structures. Starting with the known three-dimensional structure of ribonuclease T1, the observed effects of the isotope labeling are discussed on the basis of transition dipole coupling between the (12)C[double bond]O and (13)C[double bond]O groups. The experimental results were confirmed by transition dipole coupling calculations of the amide I manifold of the labeled and unlabeled variant.


Development of tolerance to the side effects of primidone.
Side effects and antiepileptic drug levels were measured during initiation of and chronic treatment with primidone (PRM). A significant change in the toxicity/serum level ratio was observed 6 h after the initial PRM dose. The amount of toxicity manifested by patients receiving PRM as their initial antiepileptic drug was not different from that of those patients previously treated with phenytoin (PHT) or carbamazepine (CBZ). These findings indicate rapid development of functional tolerance to side effects of PRM, which is not altered by previous exposure to PHT or CBZ. Phenobarbital pretreatment appears to produce cross-tolerance to the effects of PRM.


Interactions of the antiviral quinoxaline derivative 9-OH-B220 [2, 3-dimethyl-6-(dimethylaminoethyl)- 9-hydroxy-6H-indolo-[2, 3-b]quinoxaline] with duplex and triplex forms of synthetic DNA and RNA.
The binding of an antiviral quinoxaline derivative, 2,3-dimethyl- 6 - (dimethylaminoethyl) - 9 - hydroxy - 6H - indolo - [2,3 - b]quinoxaline (9-OH-B220), to synthetic double and triple helical DNA (poly(dA).poly(dT) and poly(dA).2poly(dT)) and RNA (poly(rA). poly(rU) and poly (rA).2poly(rU)) has been characterized using flow linear dichroism (LD), circular dichroism (CD), fluorescence spectroscopy, and thermal denaturation. When either of the DNA structures or the RNA duplex serve as host polymers a strongly negative LD is displayed, consistent with intercalation of the chromophoric ring system between the base-pairs/triplets of the nucleic acid structures. Evidence for this geometry also includes weak induced CD signals and strong increments of the fluorescence emission intensities upon binding of the drug to each of these polymer structures. In agreement with intercalative binding, 9-OH-B220 is found to effectively enhance the thermal stability of both the double and triple helical states of DNA as well as the RNA duplex. In the case of poly(dA).2poly(dT), the drug provides an unusually large stabilization of its triple helical state; upon binding of 9-OH-B220 the triplex-to-duplex equilibrium is shifted towards higher temperature by 52.5 deg. C in a 10 mM sodium cacodylate buffer (pH 7.0) containing 100 mM NaCl and 1 mM EDTA. When triplex RNA serves as host structure, LD indicates that the average orientation angle between the drug chromophore plane and the helix axis of the triple helical RNA is only about 60 to 65 degrees. Moreover, the thermal stabilizing capability, as well as the fluorescence increment, CD inducing power and perturbations of the absorption envelope, of 9-OH-B220 in complex with the RNA triplex are all less pronounced than those observed for the complexes with DNA and duplex RNA. These features indicate binding of 9-OH-B220 in the wide and shallow minor groove of poly(rA).2poly(rU). Based on the present results, some implications for the applications of this low-toxic, antiviral and easily administered drug in an antigene strategy, as well as its potential use as an antiretroviral agent, are discussed.


Quality evaluation of Hypericum ascyron extract by two-dimensional high-performance liquid chromatography coupled with the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.
In this paper, a heart-cutting two-dimensional high-performance liquid chromatography coupled with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was established for controlling the quality of different batches of Hypericum ascyron extract for the first time. In comparison with the common one-dimensional fingerprint, the second-dimensional fingerprint compiled additional spectral data and was hence more informative. The quality of H. ascyron extract was further evaluated by similarity measures and the same results were achieved, the correlation coefficients of the similarity of ten batches of H. ascyron extract were >0.99. Furthermore, we also evaluated the quality of the ten batches of H. ascyron extract by antibacterial activity. The result demonstrated that the quality of the ten batches of H. ascyron extract was not significantly different by MTT. Finally, we demonstrated that the second-dimensional fingerprint coupled with the MTT method was a more powerful tool to characterize the quality of samples of batch to batch. Therefore the proposed method could be used to comprehensively conduct the quality control of traditional Chinese medicines.


Interactions between the Tetrasodium Salts of EDTA and 1-Hydroxyethane 1,1-Diphosphonic Acid with Sodium Hypochlorite Irrigants.
A clinically useful all-in-one endodontic irrigant with combined proteolytic and decalcifying properties is still elusive. In this study, the chemical effects of dissolving the tetrasodium salts of 1-hydroxyethane 1,1-diphosphonic acid (Na4HEDP) or Na4EDTA directly in sodium hypochlorite (NaOCl) irrigants in polypropylene syringes were assessed during the course of 1 hour. The solubility of the salts in water was determined. Their compatibility with 1% and 5% NaOCl was measured by iodometric titration and in a calcium complexation experiment by using a Ca2+-selective electrode. The salts dissolved within 1 minute. The dissolution maximum of Na4HEDP in water (wt/total wt) was 44.6% ± 1.6%. The corresponding dissolution maximum of Na4EDTA was 38.2% ± 0.8%. Na4HEDP at 18% in 5% NaOCl caused a mere loss of 16% of the initially available chlorine during 1 hour. In contrast, a corresponding mixture between NaOCl and the Na4EDTA salt caused 95% reduction in available chlorine after 1 minute. Mixtures of 3% Na4EDTA with 1% NaOCl were more stable, but only for 30 minutes. Na4HEDP lost 24% of its calcium complexation capacity after 60 minutes. The corresponding loss for Na4EDTA was 34%. The compatibility and solubility of particulate Na4HEDP with/in NaOCl solutions are such that these components can be mixed and used for up to 1 hour. In contrast, short-term compatibility of the Na4EDTA salt with NaOCl solutions was considerably lower, decreasing at higher concentrations of either compound. Especially for Na4HEDP but also for Na4EDTA, the NaOCl had little effect on calcium complexation.


Patterns of recurrence and treatment in male breast cancer: A clue to prognosis?
Male breast cancer (MBC) patients seem to have inferior survival compared to female (FBC) ones, which is not fully explained by usual prognostic factors. Recurrence analysis could show differences in relapse patterns and/or in patients' approaches that justify these outcomes. Retrospective analysis of MBC patients treated in a cancer center between 1990 and 2014, looking for relapse. For each patient, three matched FBC patients were selected by: diagnosis' year, age (within 5 years), stage and tumors' type (only luminal-like were considered). Differences between cohorts were assessed by χ(2) test and hierarchical clustering was performed to define subgroups according to relapse local. Survival curves were calculated by Kaplan-Meier and compared using log-rank test. Statistical significance was defined as p < 0.05. Groups were balanced according to age, histological grade, stage, expression of hormonal receptors and adjuvant treatments. Median time to recurrence was equivalent, p = 0.72, with the majority of patients presented with distant metastases, p = 0.69, with more lung involvement in male, p = 0.003. Male patients were more often proposed to symptomatic treatment (21.1% vs. 4.4%, p = 0.02). Overall and from recurrence survivals were poorer for male, median: 5 years [95% confidence interval (CI): 4.1-5.9 years] and 1 year (95% CI: 0-2.1 years) vs. 10 years (95% CI: 7.8-12.2 years) and 2 years (95% CI: 1.6-2.4 years), p < 0.001 and p = 0.004, respectively, and this tendency remained in the five cluster subgroups, that identified five patterns of relapse, p = 0.003. MBC patients had the worst survival, even after controlling important factors, namely the local of relapse. Palliative systemic treatment had favorable impact in prognosis and its frequently avoidance in male could justify the outcomes differences.


Leflunomide exposure through second trimester of pregnancy: A case report.
Leflunomide is a commonly used disease modifying antirheumatic agent. However, its use is contraindicated in pregnancy. The American College of Rheumatology (ACR) guidelines recommend discontinuing Leflunomide at least 24 months before conception. If a woman is found to be pregnant while on Leflunomide, ACR suggests close monitoring and cholestyramine washout. We describe a case of a patient with a history of juvenile idiopathic arthritis who was on Leflunomide throughout the first and second trimester of her pregnancy. A cholestyramine washout regimen was started but not completed. The patient was induced at 37 weeks of gestation due to non-reassuring fetal heart rate. She ultimately delivered a healthy baby via emergency cesarian section.


Pulsed holmium:yttrium-aluminum-garnet laser-induced liquid jet as a novel dissection device in neuroendoscopic surgery.
A pressure-driven continuous jet of water has been reported to be a feasible tool for neuroendoscopic dissection owing to its superiority at selective tissue dissection in the absence of thermal effects. With respect to a safe, accurate dissection, however, continuous water flow may not be suitable for intraventricular use. The authors performed experiments aimed at solving problems associated with continuous flow by using a pulsed holmium:yttrium-aluminum-garnet (Ho:YAG) laser-induced liquid jet (LILJ). They present this candidate neuroendoscopic LILJ dissection system, having examined its mechanical characteristics and evaluated its controllability both in a tissue phantom and in a rabbit cadaveric ventricle wall. The LILJ generator was incorporated into the tip of a No. 4 French catheter so that the LILJ could be delivered via a neuroendoscope. Briefly, the LILJ was generated by irradiating an internally supplied column of physiological saline with a pulsed Ho:YAG laser (pulse duration time 350 microsec; laser energy 250-700 mJ/pulse) within a No. 4 French catheter (internal diameter 1 mm) and ejecting it from a metal nozzle (internal diameter 100 microm). The Ho:YAG laser energy pulses were conveyed by an optical fiber (core diameter 400 microm) at 3 Hz, whereas physiological saline (4 degrees C) was supplied at a rate of 40 ml/hour. The mechanical characteristics of the pulsed LILJ were investigated using high-speed photography and pressure measurements; thermal effects and controllability were analyzed using an artificial tissue model (10% gelatin of 1 mm thickness). Finally, the ventricle wall of a rabbit cadaver was dissected using the LILJ. Jet pressure increased in accordance with laser energy from 0.1 to 2 bar; this translated into a penetration depth of 0.08 to 0.9 mm per shot in the ventricle wall of the rabbit cadaver. The gelatin phantom could be cut into the desired shape without significant thermal effects and in the intended manner, with a good surgical view. The present results show that the pulsed LILJ has the potential to become a safe and reliable dissecting method for endoscopic procedures.


Cheminformatics Modeling of Amine Solutions for Assessing their CO2 Absorption Properties.
As stricter regulations on CO2 emissions are adopted worldwide, identifying efficient chemical processes to capture and recycle CO2 is of critical importance for industry. The most common process known as amine scrubbing suffers from the lack of available amine solutions capable of capturing CO2 efficiently. Tertiary amines characterized by low heats of reaction are considered good candidates but their absorption properties can significantly differ from one analogue to another despite high structural similarity. Herein, after collecting and curating experimental data from the literature, we have built a modeling set of 41 amine structures with their absorption properties. Then we analyzed their chemical composition using molecular descriptors and non-supervised clustering. Furthermore, we developed a series of quantitative structure-property relationships (QSPR) to assess amines' CO2 absorption properties from their structural characteristics. These models afforded reasonable prediction performances (e. g., Q2LOO =0.63 for CO2 absorption amount) even though they are solely based on 2D chemical descriptors and individual machine learning techniques (random forest and neural network). Overall, we believe the chemical analysis and the series of QSPR models presented in this proof-of-concept study represent new knowledge and innovative tools that could be very useful for screening and prioritizing hypothetical amines to be synthesized and tested experimentally for their CO2 absorption properties.


Epigenetic enhancement of antigen processing and presentation promotes immune recognition of tumors.
Histone deacetylase inhibitors (HDACi) have been hailed as a powerful new class of anticancer drugs. The HDACi, trichostatin A (TSA), is thought to interfere with epigenetic control of cell cycle progression in G1 and G2-M phase, resulting in growth arrest, differentiation, or apoptosis. Here, we describe a novel mechanism of action of HDACis in promoting immune responses against tumors. We report that treatment of carcinoma cells with TSA increases the expression of many components of the antigen processing machinery, including TAP-1, TAP-2, LMP-2, and Tapasin. Consistent with this result, we found that treatment of metastatic carcinoma cells with TSA also results in an increase in MHC class I expression on the cell surface that functionally translates into an enhanced susceptibility to killing by antigen-specific CTLs. Finally, we observed that TSA treatment suppresses tumor growth and increases tap-1 promoter activity in TAP-deficient tumor cells in vivo. Intriguingly, this in vivo anti-tumoral effect of TSA is entirely mediated by an increase in immunogenicity of the tumor cells, as it does not occur in immunodeficient mice. These novel insights into the molecular mechanisms controlling tumor immune escape may help revise immunotherapeutic modalities for eradicating cancers.


Scoring a backstage pass: mechanisms of ciliogenesis and ciliary access.
Cilia are conserved, microtubule-based cell surface projections that emanate from basal bodies, membrane-docked centrioles. The beating of motile cilia and flagella enables cells to swim and epithelia to displace fluids. In contrast, most primary cilia do not beat but instead detect environmental or intercellular stimuli. Inborn defects in both kinds of cilia cause human ciliopathies, diseases with diverse manifestations such as heterotaxia and kidney cysts. These diseases are caused by defects in ciliogenesis or ciliary function. The signaling functions of cilia require regulation of ciliary composition, which depends on the control of protein traffic into and out of cilia.


Labor Epidural Analgesia: Comparison of Two Different Intermittent Bolus Regimes.
Optimal labor analgesia can be provided with epidural by addition of opioid to the local anesthetic. The aim of this study is to compare the efficacy of labor epidural bolus regimes 20 mL of 0.1% ropivacaine with 40 μg fentanyl versus 15 mL of 0.1% ropivacaine with 15 μg fentanyl as epidural bolus dose. This was prospective double-blinded randomized study. After approval from the Institutional Ethical Committee, 50 consenting parturients in active labor were allotted into two groups by closed envelope technique. Group A received 20 mL of 0.1% ropivacaine with 40 μg fentanyl, whereas Group B received 15 mL of 0.1% ropivacaine with 15 μg fentanyl as an epidural bolus dose. The onset, duration of analgesia, motor block, top-up doses required, consumption of ropivacaine, and fentanyl and fetomaternal outcome were compared. Numerical variables are expressed as a mean and standard deviation and categorical variables are expressed as frequency and percentages. To obtain the association between categorical variables and different doses Fischer's exact test was applied. To compare clinical parameters between different drug doses independent two-sample t-test were applied. Mann-Whitney U-test applied for nonparametric data. Results: Effective labor analgesia with no motor blockade was observed in both groups with no failure rate. Duration of analgesia was significantly longer in Group A (166.8 ± 54.64 vs. 100.2 ± 32.39 min P < 0.001). The onset of analgesia was faster in Group A (88% vs. 16% within 7 min, P < 0.001). Labor epidural analgesia with larger volume boluses produces faster onset and prolonged duration of analgesia.


[The ultrastructural features of sputum deposition and its value in the diagnosis of pulmonary alveolar proteinosis].
To investigate the ultrastructural features of sputum deposition (SD) and its value in the diagnosis of pulmonary alveolar proteinosis (PAP). Seven patients with PAP diagnosed by lung biopsy and cytology were enrolled in this study. The patients consisted of 5 men and 2 women, whose median age was 48 years (range 36 to 73). SD and bronchoalveolar lavage fluid (BALF) sediment were made into ultrathin sections and observed under transmission electron microscope (TEM), respectively. Seven cases of control group composed of 4 men and 3 women whose median age was 49 years (range 39 to 68) including 3 cases of bacterial pneumonia, two cases of COPD and 2 cases of exudative pulmonary tuberculosis. Each SD was made into ultrathin section, and compared with the experimental group. In PAP group, Periodic acid-schiff (PAS) staining was performed on 7 sputum smears and none of them was tested positive for any components with diagnostic interest. Four cases from the 7 paraffin-embed sections of BALF sediment by microscopic examination suggested PAP. Under TEM, BALF sediment showed that many lamellar bodies existed in and outside alveolar epithelial cells, and 5 specimens were consistent with PAP diagnosis. Compared with BALF sediment, SD had apparent degeneration with more myelin phagosomes in the cytoplasm of macrophages, more lamellar bodies in alveolar epithelial cells, and lots of lamellar bodies in the shape of concentric circle in the extracellular spaces. Four from the 7 SD samples were consistent with the diagnosis of PAP. No significant difference was found between SD and BALF in the diagnosis of PAP by electronic examination (P > 0.05). In the 7 cases of control group no drifting osmiophilic lamellar bodies in extracellular space were detected. The osmiophilic lamellar bodies with diagnostic value were found in SD and BALF of patients with PAP. TEM of SD in combination with clinical manifestations and radiologic findings can make a definitive diagnosis of PAP, especially for those patients who have contraindications to lung biopsy and lung lavage.


Moral dilemmas involving anthropological and ethical dimensions in healthcare curriculum.
Currently a variety of novel scenarios have appeared within nursing practice such as confidentiality of a patient victim of abuse, justice in insolvent patients, poorly informed consent delivery, non-satisfactory medicine outputs, or the possibility to reject a recommended treatment. These scenarios presuppose skills that are not usually acquired during the degree. Thus, the implementation of teaching approaches that promote the acquisition of these skills in the nursing curriculum is increasingly relevant. The article analyzes an academic model which integrates in the curriculum a series of specific theoretical concepts together with practical skills to acquire the basic ethic assessment competency. The project includes designing two subjects, General Anthropology and Ethics-Bioethics, with an applied approach in the nursing curriculum. The sequential structure of the curriculum in both subjects is constituted by three learning domains (theoretical, practical, and communicative) with different educational strategies. No significant ethical considerations as this is a discussion paper. The model was structured from the anthropology's concepts and decision-making process, applied to real situations. The structure of the three domains theoretical-practical-communicative is present in each session. It is observed that theoretical domain fosters the capacity for critical analysis and subsequent ability to judge diverse situations. The practical domain reflected two significant difficulties: students' resistance to internalizing moral problems and the tendency to superficial criticism. The communicative domain has frequently shown that the conflicting points are in the principles to be applied. We conclude that this design achieves its objectives and may provide future nursing professionals with ethical competences especially useful in healthcare practice. The three domains of the presented scheme are associated with the same process used in decision making at individual levels, where the exercise of clinical prudence acquires particular relevance.


Administration of tranexamic acid to patients undergoing surgery for adolescent idiopathic scoliosis evokes pain and increases the infusion rate of remifentanil during the surgery.
We recently reported that tranexamic acid (TXA) evokes pain in rats by inhibiting γ-aminobutyric acid and glycine receptors on neurons in the spinal dorsal horn. Although TXA is commonly used to reduce perioperative blood loss during various surgeries, its potential to induce intraoperative nociception, thereby increasing the need for more analgesics during surgery, has not been investigated. Therefore, this study aimed to investigate whether TXA evokes pain and increases the need for a higher infusion rate of remifentanil in patients undergoing surgery for adolescent idiopathic scoliosis (AIS). Data were collected from patients with AIS who underwent posterior spinal fusion surgery from January 2008 to December 2015. All surgical procedures were performed under total intravenous anesthesia with propofol and remifentanil, by the same team of orthopedic surgeons and anesthesiologists at a single institution. Patients in the TXA group were administered TXA (loading and maintenance doses, 1000 mg and 100 mg/h) whereas those in the control group were not. Our primary outcome was the infusion rate of the intraoperative opioid analgesic remifentanil. The final analysis was based on data collected from 33 and 30 patients in the control and TXA groups, respectively. No differences were observed in the demographic data or the hemodynamic parameters between the two groups of patients. In the TXA group, the durations of surgery and anesthesia were shorter, intravascular fluid volume and total blood loss were lower, and the doses of fentanyl and ketamine administered were higher than they were in the control group (P < 0.05 for all). The mean infusion rate of intraoperative remifentanil was significantly higher in the TXA group than in the control group (control group: 0.23 ± 0.04 μg/kg/min; TXA group: 0.28 ± 0.12 μg/kg/min; P = 0.014). Patients who received TXA during the AIS surgery required a higher infusion rate of remifentanil, indicating that TXA evoked pain during the surgery.


An Overlapping Case of IgG4-related Disease and Klinefelter Syndrome with Lupus-like Serological and Neurological Features: A Case Report and Literature Review.
A 46-year-old man with Klinefelter syndrome (KS) presented with obliterative phlebitis of the lower legs with a deteriorated renal function, and elevated serum alkaline phosphatase and ataxia levels. Examinations demonstrated tubulointerstitial nephritis, obliterative phlebitis and lymphadenopathy with IgG4+ plasma cell infiltrate and sclerosing cholangitis. Although the serological profile and central nerve system involvement were compatible for systemic lupus erythematosus (SLE), a definite diagnosis of SLE was difficult to make. IgG4-related disease (IgG4-RD) with KS was finally diagnosed, and high dose prednisolone with intravenous cyclophosphamide was initiated and thereafter the patient demonstrated a prompt improvement. This is the first known case demonstrating overlapping IgG4-RD with lupus-like serological and neurological features in a patient with KS, thus highlighting the pathogenic role with the genomic background for IgG4-RD and SLE.


Study on a New Electromagnetic Flow Measurement Technology Based on Differential Correlation Detection.
Under the conditions of low flow rate and strong noise, the current electromagnetic flowmeter (EMF) cannot satisfy the requirement for measurement or separate the actual flow signal and interference signal accurately. Correlation detection technology can reduce the bandwidth and suppress noise effectively using the periodic transmission of signal and noise randomness. As for the problem that the current anti-interference technology cannot suppress noise effectively, the noise and interference of the electromagnetic flowmeter were analyzed in this paper, and a design of the electromagnetic flowmeter based on differential correlation detection was proposed. Then, in order to verify the feasibility of the electromagnetic flow measurement system based on differential correlation, an experimental platform for the comparison between standard flow and measured flow was established and a verification experiment was carried out under special conditions and with flow calibration measurements. Finally, the data obtained in the experiment were analyzed. The research result showed that an electromagnetic flowmeter based on differential correlation detection satisfies the need for measurement completely. The lower limit of the flow rate of the electromagnetic flowmeter based on the differential correlation principle could reach 0.084 m/s. Under strong external interferences, the electromagnetic flowmeter based on differential correlation had a fluctuation range in output value of only 10 mV. This shows that the electromagnetic flowmeter based on the differential correlation principle has unique advantages in measurements taken under the conditions of strong noise, slurry flow, and low flow rate.


Cloning and prokaryotic expression of WRKY48 from Caragana intermedia.
Caragana intermedia (C. intermedia) is a kind of drought-tolerant leguminous shrub. WRKY transcription factors are one of the largest family of transcription factors in plants and play critical regulatory roles in stress tolerance and the development of plants. In our study, CiWRKY48 was cloned from C. intermedia, analyzed using bioinformatics software, and expressed with a prokaryotic expression system. The results showed that the open reading frame (ORF) of CiWRKY48 was 1158bp, the molecular weight (MW) was 42 kDa, and its subcellular localization was in the nucleus. Additionally, fusion protein was obtained, and confirmed by western blotting. The stress resistance of the pET30a-His-MBP-CiWRKY48 transformed Escherichia coli expression strain was reduced under mannitol and salt treatment, compared with the control. Overall, our findings provided a foundation for uncovering the function of CiWRKY48.


Gut microbiota and microbiota-derived metabolites in cardiovascular diseases.
Cardiovascular diseases, including heart failure, coronary artery disease, atherosclerosis, aneurysm, thrombosis, and hypertension, are a great economic burden and threat to human health and are the major cause of death worldwide. Recently, researchers have begun to appreciate the role of microbial ecosystems within the human body in contributing to metabolic and cardiovascular disorders. Accumulating evidence has demonstrated that the gut microbiota is closely associated with the occurrence and development of cardiovascular diseases. The gut microbiota functions as an endocrine organ that secretes bioactive metabolites that participate in the maintenance of cardiovascular homeostasis, and their dysfunction can directly influence the progression of cardiovascular disease. This review summarizes the current literature demonstrating the role of the gut microbiota in the development of cardiovascular diseases. We also highlight the mechanism by which well-documented gut microbiota-derived metabolites, especially trimethylamine N-oxide, short-chain fatty acids, and phenylacetylglutamine, promote or inhibit the pathogenesis of cardiovascular diseases. We also discuss the therapeutic potential of altering the gut microbiota and microbiota-derived metabolites to improve or prevent cardiovascular diseases.


Transformation of an Amine Moiety of Atenolol during Water Treatment with Chlorine/UV: Reaction Kinetics, Products, and Mechanisms.
Transformation of atenolol (ATN), a micropollutant containing a secondary (2°) amine moiety, can be significantly enhanced in water treatment with sequential and combined use of chlorine and UV (chlorine/UV) through photolysis of the N-Cl bond. This study investigated the transformation kinetics, products, and mechanisms of the amine moiety of ATN in chlorine/UV (254 nm). The fluence-based, photolysis rate constant for N-Cl ATN was 2.0 × 10-3 cm2/mJ. Transformation products (TPs) with primary (1°) amines were mainly produced, but TPs with 2° and 3° amines were also formed, on the basis of liquid chromatography (LC)/quadrupole-time-of-flight/mass spectrometry and LC/UV analyses. The amine-containing TPs could be further transformed in chlorine/UV (with residual chlorine in post UV) via formation and photolysis of new N-Cl bonds. Photolysis of N-Cl 1° amine TPs produced ammonia as a major product. These data could be explained by a reaction mechanism in which the N-Cl bond was cleaved by UV, forming aminyl radicals that were transformed via 1,2-hydrogen shift, β-scission, intramolecular addition, and 1,2-alkyl shift. Among these, the 1,2-alkyl shift is newly discovered in this study. Despite enhanced transformation, only partial mineralization of the ATN's amine moiety was expected, even under chlorine/UV advanced oxidation process conditions. Overall, the kinetic and mechanistic information from this study can be useful for predicting the transformation of amine moieties by chlorine/UV water treatment.


Nitrogen starvation and TorC1 inhibition differentially affect nuclear localization of the Gln3 and Gat1 transcription factors through the rare glutamine tRNACUG in Saccharomyces cerevisiae.
A leucine, leucyl-tRNA synthetase-dependent pathway activates TorC1 kinase and its downstream stimulation of protein synthesis, a major nitrogen consumer. We previously demonstrated, however, that control of Gln3, a transcription activator of catabolic genes whose products generate the nitrogenous precursors for protein synthesis, is not subject to leucine-dependent TorC1 activation. This led us to conclude that excess nitrogen-dependent down-regulation of Gln3 occurs via a second mechanism that is independent of leucine-dependent TorC1 activation. A major site of Gln3 and Gat1 (another GATA-binding transcription activator) control occurs at their access to the nucleus. In excess nitrogen, Gln3 and Gat1 are sequestered in the cytoplasm in a Ure2-dependent manner. They become nuclear and activate transcription when nitrogen becomes limiting. Long-term nitrogen starvation and treatment of cells with the glutamine synthetase inhibitor methionine sulfoximine (Msx) also elicit nuclear Gln3 localization. The sensitivity of Gln3 localization to glutamine and inhibition of glutamine synthesis prompted us to investigate the effects of a glutamine tRNA mutation (sup70-65) on nitrogen-responsive control of Gln3 and Gat1. We found that nuclear Gln3 localization elicited by short- and long-term nitrogen starvation; growth in a poor, derepressive medium; Msx or rapamycin treatment; or ure2Δ mutation is abolished in a sup70-65 mutant. However, nuclear Gat1 localization, which also exhibits a glutamine tRNACUG requirement for its response to short-term nitrogen starvation or growth in proline medium or a ure2Δ mutation, does not require tRNACUG for its response to rapamycin. Also, in contrast with Gln3, Gat1 localization does not respond to long-term nitrogen starvation. These observations demonstrate the existence of a specific nitrogen-responsive component participating in the control of Gln3 and Gat1 localization and their downstream production of nitrogenous precursors. This component is highly sensitive to the function of the rare glutamine tRNACUG, which cannot be replaced by the predominant glutamine tRNACAA. Our observations also demonstrate distinct mechanistic differences between the responses of Gln3 and Gat1 to rapamycin inhibition of TorC1 and nitrogen starvation.


Detection of high-risk groups and individuals for periodontal diseases. Clinical assessment of the periodontium.
The fundamental concepts of measuring periodontal diseases and the interpretation of such information as an historical record of disease, rather than disease activity, emphasises the need for improved diagnostic and prognostic tests. Criteria for an indicator of disease activity were suggested and an index fulfilling these should allow sites to be categorised as "active", quiescent or healing, and enable one to predict the risk of future disease activity. The ability of current measurements and indices, routinely used during clinical assessments of periodontal diseases, to fulfill the suggested criteria was considered and rejected in all cases. It is concluded that clinical parameters are only capable of identifying disease retrospectively, indicating the need for longitudinal, rather than cross-sectional studies in the search for clinical and laboratory markers of disease activity and susceptibility.


Ultrafast photoinduced dynamics of pigment yellow 101: fluorescence, excited-state intramolecular proton transfer, and isomerization.
The ultrafast excited-state dynamics of the fluorescent pigment yellow 101 (P.Y.101) and the closely related 1,1'-naphthalazine, a nonfluorescent derivative that lacks the OH groups at the naphthyl rings, are studied combining femtosecond spectroscopy and high-level quantum chemical calculations. The observed ultrafast dynamics and the spectral signature of photoexcited 1,1'-naphthalzine can be consistently explained with a previously proposed mechanism, suggesting fluorescence quenching via an optically forbidden npi* state. In contrast, for a description of the excited-state dynamics of P.Y.101, the expected simple absorption/fluorescence model is not adequate. Instead, besides fluorescence as the main decay channel of the excited-state population, ultrafast excited-state intramolecular proton transfer (ESIPT) and isomerization processes have to be considered for a complete understanding of the complex subnanosecond dynamics. Combining experiment and theory, the following kinetic model is derived: upon photoexcitation a major part of the excited-state population decays via fluorescence from an enol-type isomer of P.Y.101, while a small part of the population undergoes ESIPT and fluoresces from a keto-type form. Furthermore, arguments are given that, to a minor extent, also trans-cis isomerization of the keto form takes place on the S1 surface leading probably to a long-lived cis-keto form in the ground state. The remarkable photostability of this organic pigment is thus achieved by the interplay of different ultrafast nondestructive decay channels.


A mitogenic lectin from Rhizoctonia bataticola arrests growth, inhibits metastasis, and induces apoptosis in human colon epithelial cancer cells.
The correlation between colorectal cancer (CRC) progression and altered expression of N-glycans can be considered in search for new biomarkers and anticancer agents to control CRC. Earlier N-glycan specific mitogenic lectin from Rhizoctonia bataticola (RBL) has been reported which has growth inhibitory and apoptotic effect on human ovarian and leukemic cells, but mitogenic effect on normal PBMCs revealing its clinical potential. Here, we report the effect of RBL on human colon cancer HT 29, SW480, and SW620 cell growth and its differential binding to human normal colon and cancer tissues. RBL has strong binding to both primary and metastatic colon cancer cells with MFI of 403, 404, and 192, respectively for HT 29, SW480, and SW620 cells. RBL shows dose and time dependent growth inhibitory effect with IC50 of 5, 6.4, and 6.8 µg/mL, respectively for HT 29, SW480, and SW620 cells. RBL inhibited the clonogenicity of colon carcinoma cells. RBL arrests metastatic SW620 cell growth at S phase, increased hypodiploid population by 6.1%, 14.3%, and 23.2%, respectively at 12, 24, and 36 h. Further, RBL induces SW620 cell apoptosis in time dependent manner, showed increased release of ROS and nuclear degradation compared to lectin untreated control. Adhesion, wound healing, invasion, and migration assays demonstrated anti-metastasis effect of RBL in SW620 cells apart from its growth inhibitory effect. Anti angiogenic effect of RBL was demonstrated by CAM assay. All these results show the promising potential of RBL both as diagnostic and therapeutic agent.


Cell cycle dysregulation by human cytomegalovirus: influence of the cell cycle phase at the time of infection and effects on cyclin transcription.
Human cytomegalovirus (HCMV) infection inhibits cell cycle progression and alters the expression of cyclins E, A, and B (F. M. Jault, J.-M. Jault, F. Ruchti, E. A. Fortunato, C. Clark, J. Corbeil, D. D. Richman, and D. H. Spector, J. Virol. 69:6697-6704, 1995). In this study, we examined cell cycle progression, cyclin gene expression, and early viral events when the infection was initiated at different points in the cell cycle (G0, G1, and S). In all cases, infection led to cell cycle arrest. Cells infected in G0 or G1 phase also showed a complete or partial absence, respectively, of cellular DNA synthesis at a time when DNA synthesis occurred in the corresponding mock-infected cells. In contrast, when cells were infected near or during S phase, many cells were able to pass through S phase and undergo mitosis prior to cell cycle arrest. S-phase infection also produced a delay in the appearance of the viral cytopathic effect and in the synthesis of immediate-early and early proteins. Labeling of cells with bromodeoxyuridine immediately prior to HCMV infection in S phase revealed that viral protein expression occurred primarily in cells which were not engaged in DNA synthesis at the time of infection. The viral-mediated induction of cyclin E, maintenance of cyclin-B protein levels, and inhibitory effects on the accumulation of cyclin A were not significantly affected when infection occurred during different phases of the cell cycle (G0, G1, and S). However, there was a delay in the observed inhibition of cyclin A in cells infected during S phase. This finding was in accord with the pattern of cell cycle progression and delay in viral gene expression associated with S-phase infection. Analysis of the mRNA revealed that the effects of the virus on cyclin E and cyclin A, but not on cyclin B, were primarily at the transcriptional level.


Membrane sialic acid on target particles modulates their phagocytosis by a trypsin-sensitive mechanism on human monocytes.
Monolayers of human peripheral blood monocytes in the absence of exogenous proteins ingest a variety of natural particulate activators of the human alternative complement pathway. Sheep erythrocytes, which do not ordinarily activate the human alternative complement pathway or initiate a direct monocyte phagocytic response, can be modified to exhibit both functions by the deletion or alteration of membrane sialic acid residues. Enzymatic removal of the sialic acid residues with sialidase or their conversion to heptulosonic acid derivatives by limited oxidation with NaIO4 and reduction with BH4- have equivalent dose-response effects on the capacity of the altered sheep erythrocytes to initiate the phagocytic response by human monocytes or to activate the alternative pathway in human serum. The deposition of C3b on native sheep erythrocytes had little effect on their ingestion by human monocytes, whereas the fixation of C3b on desialated sheep erythrocytes had a synergistic effect on the percentage of monocytes ingesting such a particle. The monocyte receptor essential for ingestion of desialated sheep erythrocytes or desialated sheep erythrocytes bearing C3b was inactivated by concentrations of trypsin that also prevented the monocytes from ingesting natural activators of the human alternative complement pathway, but did not alter the receptors for C3b or the Fc portion of IgG. The capacity of the nonimmune host to respond to desialated particles by initiating the monocyte ingestive process and by activating the alternative complement pathway to provide the synergy afforded by C3b deposition on that particle represents a primitive biochemical basis for differentiation of nonself from self.


[Gender dysphoria in children and adolescents].
Despite rising demand for treatment, far too few specialist services for gender dysphoric children and adolescents exist. Due to complex issues related to indication of physical treatment with lifelong consequences a thorough interdisciplinary treatment service with development-related evaluation is necessary. The aim of this paper is to outline the professional and social tension, in which the practitioner works, as well as to provide insight into the diversity of developmental trajectories and difficult decisions or dilemmas.


SGLT2 inhibitors to control glycemia in type 2 diabetes mellitus: a new approach to an old problem.
Sodium-glucose cotransporter 2 (SGLT2) inhibitors are a new class of antidiabetic agents with a novel insulin-independent mechanism of action. The SGLT2 is a transporter found in the proximal tubule of the kidney and is responsible for approximately 90% of renal glucose reabsorption. The SGLT2 inhibitors reduce reabsorption of glucose in the kidney, resulting in glucose excretion in the urine (50-90 g of ~180 g filtered by the kidneys daily), which in turn lowers plasma glucose levels in people with diabetes. The insulin-independent mechanism of action of SGLT2 inhibitors dictates that they are associated with a very low risk of hypoglycemia and can be used in patients with any degree of β-cell function or insulin sensitivity. Clinical trials have shown that SGLT2 inhibitors are effective at reducing blood glucose levels, body weight, and blood pressure when used as monotherapy or in combination with other antidiabetic agents in patients with type 2 diabetes mellitus. Treatment with SGLT2 inhibitors is generally well tolerated, although these agents have been associated with an increased incidence of genital infections. The SGLT2 inhibitors have become a valuable addition to the armory of drugs used to treat patients with type 2 diabetes mellitus, and several agents within the class are currently under investigation in phase III clinical trials.


Use of translesional coronary flow velocity for interventional decisions in a patient with multiple intermediately severe coronary stenoses.
Translesional pressure gradients are currently not performed routinely for most angioplasty procedures because of the technical limitations. With the use of an 0.18 inch Doppler flow velocity guidewire, coronary flow velocity, both proximal and distal to a lesion, can be easily assessed. In branching arteries, significant lesions are characterized by a ratio of proximal to distal flow velocity of > 1.7, loss of the normal phasic diastolic predominant flow velocity pattern, and/or loss of distal hyperemia. We describe the use of coronary flow velocity in assisting important decision making in a young patient with a recent myocardial infarction and multiple coronary lesions. The decision for angioplasty of the hemodynamically significant stenosis was confirmed by translesional flow velocity measurements. A rational approach to coronary intervention in patients with multiple stenoses of intermediate severity appears to be facilitated by direct measurement of translesional flow dynamics.


Dural Extranodal Marginal Zone Lymphoma in an XRCC2 Mutation Carrier.
Dural extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) is a rare entity without an associated recurrent genetic abnormality. Only one case has been described in a woman with history of breast carcinoma without a known genetic predisposition. Here, we report a case of a 56-year-old woman heterozygous for XRCC2 mutation with a history of Graves' disease and bilateral breast carcinomas, who was found to have a diffusely infiltrative extra-axial mass in the high parietal convexity with infiltration into the adjacent superior sagittal sinus. The morphologic, immunophenotypic, and molecular findings were diagnostic of MALT lymphoma. Staging bone marrow demonstrated involvement by the neoplasm. Although the study was limited to only the clinically significant laboratory evaluation, it may serve as an important addition to the current knowledge of the pathogenic potential of a loss of function mutation in this rarely reported cancer predisposition gene.


Widespread association between the ericoid mycorrhizal fungus Rhizoscyphus ericae and a leafy liverwort in the maritime and sub-Antarctic.
A recent study identified a fungal isolate from the Antarctic leafy liverwort Cephaloziella varians as the ericoid mycorrhizal associate Rhizoscyphus ericae. However, nothing is known about the wider Antarctic distribution of R. ericae in C. varians, and inoculation experiments confirming the ability of the fungus to form coils in the liverwort are lacking. Using direct isolation and baiting with Vaccinium macrocarpon seedlings, fungi were isolated from C. varians sampled from eight sites across a 1875-km transect through sub- and maritime Antarctica. The ability of an isolate to form coils in aseptically grown C. varians was also tested. Fungi with 98-99% sequence identity to R. ericae internal transcribed spacer (ITS) region and partial large subunit ribosomal (r)DNA sequences were frequently isolated from C. varians at all sites sampled. The EF4/Fung5 primer set did not amplify small subunit rDNA from three of five R. ericae isolates, probably accounting for the reported absence of the fungus from C. varians in a previous study. Rhizoscyphus ericae was found to colonize aseptically-grown C. varians intracellularly, forming hyphal coils. This study shows that the association between R. ericae and C. varians is apparently widespread in Antarctica, and confirms that R. ericae is at least in part responsible for the formation of the coils observed in rhizoids of field-collected C. varians.


Adenoviral gene transfer of hepatic stimulator substance confers resistance against hepatic ischemia-reperfusion injury by improving mitochondrial function.
Hepatic stimulator substance (HSS) has been suggested to protect liver cells from various toxins. However, the precise role of HSS in hepatic ischemia-reperfusion (I/R) injury remains unknown. This study aims to elucidate whether overexpression of HSS could attenuate hepatic ischemia-reperfusion injury and its possible mechanisms. Both in vivo hepatic I/R injury in mice and in vitro hypoxia-reoxygenation (H/R) in a cell model were used to evaluate the effect of HSS protection after adenoviral gene transfer. Moreover, a possible mitochondrial mechanism of HSS protection was investigated. Efficient transfer of the HSS gene into liver inhibited hepatic I/R injury in mice, as evidenced by improvement in liver function tests, the preservation of hepatic morphology, and a reduction in hepatocyte apoptosis. HSS overexpression also inhibited H/R-induced cell death, as detected by cell viability and cell apoptosis assays. The underlying mechanism of this hepatic protection might involve the attenuation of mitochondrial dysfunction and mitochondrial-dependent cell apoptosis, as shown by the good preservation of mitochondrial ultrastructure, mitochondrial membrane potential, and the inhibition of cytochrome c leakage and caspase activity. Moreover, the suppression of H/R-induced mitochondrial ROS production and the maintenance of mitochondrial respiratory chain complex activities may participate in this mechanism. This new function of HSS expands the possibility of its application for the prevention of I/R injury, such as hepatic resection and liver transplantation in clinical practice.


An examination of product packaging marketing strategies used to promote pediatric multivitamins.
The purpose of this study was to describe the nature of marketing strategies for multivitamin and multimineral (MVM) supplement packaging and to assess the extent to which these supplements are marketed as food products. A cross-sectional study of children's supplement packaging was conducted. Descriptive statistics identified common marketing practices. Websites of the three largest retail chain pharmacies in the United States and MVM manufacturers were accessed. The study's sample consisted of packaging for 52 children's MVM supplements. Child-targeted marketing included reference to trademarked characters on 42.3% of MVM packaging (n = 22). More than 80% of the sample (n = 42) listed fruity flavors and almost all packaging included descriptive words related to the MVMs' shape and/or flavor (88.5%, n = 46). Nearly one-fifth of the packaging (n = 10) pictured a food item. With respect to parent-targeted promotional language, almost 83% of the supplement packages (n = 43) included text on the support of bodily structure/function. More than half of the sample (53.8%, n = 28) had promotional language related to dietary practice (e.g. organic, gluten-free). Pediatricians can play a role in ensuring that parents are aware of (1) possible risks associated with MVM overconsumption, and (2) the importance of deriving vitamins and minerals from a balanced diet. Given the high number of exposures to pediatric MVMs among youth and established influence of food marketing on shaping children's perceptions and behaviors, further research is necessary to determine the extent to which children's MVMs are marketed as a food product and perceived as such by children.


Radical Crotylation of Aldehydes with 1,3-Butadiene by Photoredox Cobalt and Titanium Dual Catalysis.
Metal-hydride hydrogen atom transfer (MHAT) has been recognized as a powerful method for alkene functionalization; however, photochemical MAT-mediated chemoselective functionalization of dienes remains undeveloped. In this study, we report a radical strategy (1e-) through MHAT using photoredox cobalt and titanium dual catalysis for aldehyde crotylation with butadiene, achieving excellent regio- and diastereoselectivity.


Retained Digital Flexible Ureteroscopes.
This report documents two instances of retained flexible ureteroscopes at the time of ureteroscopy and laser lithotripsy in a healthy 37-year-old male and a 53-year-old male with a pelvic kidney. We describe maneuvers to remove the ureteroscope endoscopically in the first case, while the second case required conversion to open surgery for ureteroscope extrication.


Effect of volatile fatty acid infusion on development of the rumen epithelium in neonatal sheep.
The purpose of this study was to determine whether the continuous intraruminal infusion of calculated physiological concentrations of volatile fatty acids (VFA) stimulated the metabolic development of the neonatal rumen. Eight 1-wk-old lambs were assigned to one of three treatments: saline infusion (three lambs), VFA infusion (three lambs), or no infusion (two lambs). Rumen catheters were surgically implanted into lambs in the infusion groups. The amount of VFA infused, beginning at 2 wk of age, increased weekly in equal increments of 12.5% of the estimated net energy requirement until, at 6 wk of age, lambs received 50% of their estimated net energy requirement from the infused VFA. All lambs consumed milk replacer for ad libitum intake and had free access to water. The lambs that were infused with VFA tended to have longer rumen papillae. There were no differences in width or number of papillae per square centimeter across treatments. Rumen epithelial cells isolated from lambs that were infused with VFA tended to oxidize less glucose and produce more acetoacetate than did cells from lambs that were infused with saline or from uninfused lambs. beta-Hydroxybutyrate production by isolated rumen epithelial cells and concentrations of blood glucose, acetoacetate, and beta-hydroxybutyrate were not different among the three treatments. Thus, infusion of physiological concentrations of VFA appears to stimulate some aspects of rumen metabolic development.


Development of bone-conduction mobile phones: assessment of hearing mechanisms by measuring psychological characteristics and acoustical properties in the outer ear canal.
We have been developing novel mobile phones using bone conduction, with flat-panel loudspeakers that convey speech sound by vibrating the pinna. In bone conduction via the pinna, i.e., pinna conduction, it is thought that speech sounds are conveyed via both air- and bone-conduction pathways. To obtain useful information for further development of bone-conduction mobile phones, peripheral mechanisms of the pinna conduction need to be clarified. In this study, hearing thresholds, sound field in the outer ear canals, and vibrations of the inner wall of the outer ear canals were measured while normal-hearing participants used pinna-conduction mobile phones. Thresholds decreased linearly as contact pressure increased below 1 kHz, but contact pressure did not affect thresholds above 2 kHz. Additionally, sound fields in the ipsilateral ear canal showed similar results. These results indicate that there is a considerable degree of bone-conduction components from the pinna to the inner ear, which only allow sounds below 1 kHz through. Because similar characteristics were observed in the threshold and the sound field in the outer ear canal, we suggest that osseotympanic emission, sound emission into the ear canal from the inner wall, and air conduction via external auditory foramen are the dominant components of pinna conduction. However, in the vibration measurement, differences between the ipsi- and contra-lateral responses were smaller than the sound field measurement. The smaller inter-lateral differences of the vibration in the outer ear canal suggest the existence of a significant amount of bone-conduction components that directly reach the middle or inner ear. Although the amount of such bone-conduction components does not seem sufficient for pinna.


Impact of a 25% Salt Reduction on the Microbial Load, Texture, and Sensory Attributes of a Traditional Dry-Cured Sausage.
Catalão is a Portuguese dry-cured traditional sausage, highly appreciated for its distinctive sensory properties. The aim of this study was to evaluate the impact of a 25% salt reduction on Catalão manufactured with either purebred Alentejano (Al) or crossbred Iberian × Duroc (IDr) pork meat, on its physicochemical and microbiological stability, texture parameters, and sensory attributes. No significant effect of salt reduction or genotype was observed for pH, aW, and microbiological parameters. PUFA content was significantly higher for Al Catalão, particularly due to the content in linoleic and linolenic fatty acids. IDr 3% NaCl samples had the highest mean n6/n3 PUFA ratio, and the highest mean values for the atherogenicity and thrombogenicity indices, showing that both genotype and salt content influence these nutritional indices. Texture profile of Catalão was significantly influenced by salt content and genotype. Al samples were less adhesive, cohesive, and easier to chew. Low-salt Catalão was harder, more adhesive, and less cohesive, with lower resilience and higher chewiness values. Regarding sensory attributes, salt content influenced the product aroma, with reduced-salt sausages being evaluated as significantly less aromatic. Overall, a 25% salt reduction did not have a significant impact on the quality, stability, and sensory evaluation of Catalão.


Is the retrograde axonal transport of 3H-5-HT a specific process of serotoninergic neurons?
The specificity of the retrograde axonal transport of 3H-serotonin (3H-5-HT) was radioautographically studied in the afferents to the olfactory bulb (O.B.). Injections of 3H-5-HT of different concentrations (10(-2), 10(-3), 10(-4) and 10(-5) M) were performed into the O.B. of catron pretreated rats. Following injection of 3H-5-HT (10(-2) M), a cytoplasmic perikaryal labeling was observed in the bulk of afferents to the O.B. (aminergic and non-aminergic neurons). When lower concentrations of 3H-5-HT (10(-5) M) were injected into the O.B., the retrograde labeling was only seen in the raphe dorsalis (RD) serotoninergic perikarya. The specificity of the uptake-retrograde transport of 3H-5-HT seems to depend on the selectivity of uptake by nerve terminals.


tinselR-an R Shiny Application for Annotating Phylogenetic Trees.
Public health laboratories obtain whole-genome sequences of pathogens to confirm outbreaks and identify transmission routes. Here, we present tinselR, an open-source and user-friendly application for visualization and annotation of relatedness among pathogens with phylogenetic trees. By combining annotation and phylogenetics, we simplify one critical step in the pathogen analysis workflow.


Dramatic weight loss associated with commencing clozapine.
The authors report the case of a 44-year-old man with a long history of chronic enduring schizophrenia who experienced dramatic weight loss after commencing treatment with clozapine, an antipsychotic medication characteristically associated with the greatest degree of weight gain among medical treatments for schizophrenia. He was obese with a body mass index (BMI) of 41.5 kg/m(2), but after commencing clozapine therapy he experienced an improvement in psychotic symptoms and 40% loss of his body weight attained through an altered diet and exercise regime, which resulted in him attaining a normal BMI of 24.8 kg/m(2).


Aortic stenosis severity is not a risk factor for poststenotic dilatation of the ascending aorta.
Dilatation of the ascending aorta in aortic stenosis may be partly explained by intrinsic wall structure changes, but the relative contribution of altered hemodynamics is unclear. The aim of this study was to assess the association between ascending aortic dimensions and valve stenosis severity. An analysis of echocardiographic examinations was conducted in 296 patients with aortic stenosis (179 males, mean age 71 years), 57 with bicuspid and 239 with tricuspid aortic valve, mean transaortic gradient 43+/-20 mmHg, and not more than moderate aortic regurgitation. Aortic dimensions at the level of annulus, sinuses of Valsalva, sinotubular junction and proximal ascending aorta were measured. Only height (p<0.001), degree of aortic regurgitation (p<0.01) and presence of bicuspid aortic valve (p<0.001) were independent predictors of ascending aortic dimensions. An independent association between aortic pressure gradients and proximal ascending aortic dimensions was not observed in patients with bicuspid or tricuspid aortic valve stenosis. Therefore, the poststenotic dilatation of the ascending aorta is not explained by aortic stenosis severity itself. Possible nonhemodynamic causes deserve detailed study at the time of diagnosis.


Isolation and characterization of linear polylactosamines containing one and two site-specifically positioned Lewis x determinants: WGA agarose chromatography in fractionation of mixtures generated by random, partial enzymatic alpha3-fucosylation of pure polylactosamines.
We report that isomeric monofucosylhexasaccharides, Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1- 3Galbeta1-4(Fucalpha1-3) GlcNAc, Galbeta1-4GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3) GlcNAcbeta1-3Galbeta1-4 GlcNAc and Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3Galbeta1- 4GlcNAcbeta1-3Galbeta1-4 GlcNAc, and bifucosylhexasaccharides Galbeta1-4GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3) GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3)GlcNAc, Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3Galbeta1- 4GlcNAcbeta1-3Galbeta1-4 (Fucalpha1-3)GlcNAc and Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3Galbeta1-4( Fucalpha1-3)GlcNAcbeta1-3Galbeta1-4GlcNAc can be isolated in pure form from reaction mixtures of the linear hexasaccharide Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1- 3Galbeta1-4GlcNAc with GDP-fucose and alpha1,3-fucosyltransferases of human milk. The pure isomers were characterized in several ways;1H-NMR spectroscopy, for instance, revealed distinct resonances associated with the Lewis x group [Galbeta1-4(Fucalpha1-3)GlcNAc] located at the proximal, middle, and distal positions of the polylactosamine chain. Chromatography on immobilized wheat germ agglutinin was crucial in the separation process used; the isomers carrying the fucose at the reducing end GlcNAc possessed particularly low affinities for the lectin. Isomeric monofucosyl derivatives of the pentasaccharides GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1-3Galbeta1- 4Gl cNAc and Galalpha1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4G lcN Ac and the tetrasaccharide Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAc were also obtained in pure form, implying that the methods used are widely applicable. The isomeric Lewis x glycans proved to be recognized in highly variable binding modes by polylactosamine-metabolizing enzymes, e.g., the midchain beta1,6-GlcNAc transferase (Leppänen et al., Biochemistry, 36, 13729-13735, 1997).


Association of vitamin D receptor gene polymorphisms with prostate cancer risk in the Pakistani population.
Vitamin D receptor (VDR) gene has been a subject of extensive pharmacogenetic research recently. Association studies between different types of cancers including prostate cancer (PCa) and VDR gene polymorphism have also been conducted. The objective of this study was to find possible associations between PCa and VDR gene polymorphisms in the Pakistani population. A total of 162 subjects, including prostate cancer patients and controls, were genotyped for Apa I, Taq I and Fok I polymorphisms in the VDR gene using allele specific PCR, PCR-RFLP and direct DNA sequencing. Allelic frequencies were tested for Hardy-Weinberg equilibrium and associations between the genetic markers and PCa were calculated using logistic regression. Apa I CC genotype was found to have strongest association with PCa risk, and "A" genotype was found to have protective effect. Fok I and Taq I did not have appreciable levels of association with PCa, although Taq I "TC" heterozygotes seemed to have some protective effect. Similarly the "C" allele of Fok I also seemed to have protective effect. To our knowledge, this is the first report showing association between VDR gene polymorphisms and PCa in Pakistan. Our findings may be somewhat skewed because of small sample size and tendency of consanguineous marriages in Pakistani society; nevertheless, it shows the trend of association and protective effects of certain VDR gene polymorphisms against PCa.


Low intensity vibration exposure in postmen.
The complaints of white fingers in postmen who had used motorbikes and had been exposed to low intensity vibration were studied. Their vibration exposure was found to be 1-3 m/s2 per day, four hour equivalent value of frequency weighted acceleration. The rate of white fingers among all subjects was 2.8%. The rate was related not only to vibration/cold exposure but also to age and delivery area locations. The major part of the white finger occurrence reported here was considered to be due to primary Raynaud's syndrome, and it seems safe to conclude that, a daily vibration exposure of up to 2.5-3 m/s2 (4 h equivalent value) has not caused white fingers in this population.


Complicated Rapunzel syndrome mimicking intussusception.
A 5-year-old girl presented with a 3-day history of pain and distension of abdomen, bilious vomiting, bleeding per rectum and a hard lump in the left iliac fossa. Intussusception was clinically diagnosed. On exploratory laparotomy, trichobezoar showing cast of the stomach, duodenal C-loop and tail were extracted. The stomach cast was impacted at the distal ileum, while its tail traversed the ileum, ileocecal valve and extended up to the hepatic flexor. At the site of impaction, a large ileal perforation, covered by bezoar was present. Hence, x-ray did not reveal pneumoperitoneum. There was no evidence of trichobezoar in the stomach. Perforation was exteriorised as loop ileostomy. She was of normal intelligence. Psychological evaluation of the child was performed and a behaviour therapy was advocated. Ileostomy closure was done after 2 months. At 6 months follow-up, no recurrence was found.


(Ta/Si) multilayer as a wide-bandpass monochromator material.
Specular and non-specular X-ray reflectivity intensities of a (Ta/Si)(60) multilayer sample were measured to characterize its interface structure. Since the multilayer has a good reflectance at its multilayer peaks, its performance as a wide-bandpass monochromator for X-ray scattering experiments of polymers has been tested.


[The model of perioperative risk assessment in elderly patients - interim analysis].
Demographic changes in contemporary society require implementation of proper perioperative care of elderly patients due to an increased risk of perioperative complications in this group. Preoperative assessment of health status identifies risks and enables preventive interventions, improving outcomes of surgical treatment. The Comprehensive Geriatric Assessment contains numerous diagnostic tests and consultations, which is expensive and difficult to use in everyday practice. The development of a simplified model of perioperative assessment of elderly patients will help identifying the group of patients who require further diagnostic workup. The aim of the study is to evaluate the usefulness of the tests used in a proposed model of perioperative risk assessment in elderly patients. In a group of 178 patients older than 64 years admitted for surgical procedures, a battery of tests was performed. The proposed model of perioperative risk assessment included: Charlson Comorbidity Index, ADL (activities of daily living), TUG test (timed "up and go" test), MNA (mini nutritional assessment), AMTS (abbreviated mental test score), spirometry measurement of respiratory muscle strength (Pimax, Pemax). Distribution of abnormal results of each test has been analysed. The Charlson Index over 6 points was recorded in 10.1% of patients (15.1% in cancer patients). Abnormal result of the TUG test was observed in 32.1%. The risk of malnutrition in MNA test has been identified in 29.7% (39.2% in cancer patients). Abnormal test results at the level of 10-30% indicate potential diagnostic value of Charlson Comorbidity Index, TUG test and MNA in the evaluation of perioperative risk in elderly patients.


Specific TLR-mediated HSP70 activation plays a potential role in host defense against the intestinal parasite Giardia duodenalis.
Giardia duodenalis, an important flagellated noninvasive protozoan parasite, infects the upper small intestine and causes a disease termed giardiasis globally. Few members of the heat shock protein (HSP) family have been shown to function as potential defenders against microbial pathogens, while such information is lacking for Giardia. Here we initially screened and indicated that in vitro Giardia challenge induced a marked early upregulation of HSP70 in intestinal epithelial cells (IECs). As noted previously, apoptotic resistance, nitric oxide (NO)-dependent cytostatic effect and parasite clearance, and epithelial barrier integrity represent effective anti-Giardia host defense mechanisms. We then explored the function of HSP70 in modulating apoptosis, NO release, and tight junction (TJ) protein levels in Giardia-IEC interactions. HSP70 inhibition by quercetin promoted Giardia-induced IEC apoptosis, viability decrease, NO release reduction, and ZO-1 and occludin downregulation, while the agonist celastrol could reverse these Giardia-evoked effects. The results demonstrated that HSP70 played a previously unrecognized and important role in regulating anti-Giardia host defense via attenuating apoptosis, promoting cell survival, and maintaining NO and TJ levels. Owing to the significance of apoptotic resistance among those defense-related factors mentioned earlier, we then elucidated the anti-apoptotic mechanism of HSP70. It was evident that HSP70 could negatively regulate apoptosis in an intrinsic way via direct inhibition of Apaf-1 or ROS-Bax/Bcl-2-Apaf-1 axis, and in an extrinsic way via cIAP2-mediated inhibition of RIP1 activity. Most importantly, it was confirmed that HSP70 exerted its host defense function by downregulating apoptosis via Toll-like receptor 4 (TLR4) activation, upregulating NO release via TLR4/TLR2 activation, and upregulating TJ protein expression via TLR2 activation. HSP70 represented a checkpoint regulator providing the crucial link between specific TLR activation and anti-Giardia host defense responses. Strikingly, independent of the checkpoint role of HSP70, TLR4 activation was proven to downregulate TJ protein expression, and TLR2 activation to accelerate apoptosis. Altogether, this study identified HSP70 as a potentially vital defender against Giardia, and revealed its correlation with specific TLR activation. The clinical importance of HSP70 has been extensively demonstrated, while its role as an effective therapeutic target in human giardiasis remains elusive and thus needs to be further clarified.