Real-time fingerprinting of structural isomers using laser induced breakdown spectroscopy.
Laser induced breakdown spectroscopy (LIBS) has surfaced as an attractive alternative to mass spectrometry and wet chemistry methods for chemical identification, driven by its real-time, label-free nature. Rapid analysis needs, especially in high-energy materials and pharmaceutical compounds, have further fueled an increasing number of refinements in LIBS. Yet, isomers are seldom identifiable by LIBS as they generate nearly identical spectra. Here we employ a suite of chemometric approaches to exploit the subtle, but reproducible, differences in LIBS spectra acquired from structural isomers, a set of pyrazoles, to develop a sensitive and reliable segmentation method. We also investigate the possible mechanistic principles (causation) behind such spectral variations and confirm their statistically significant nature that empowers the excellent classification performance.


Synthesis of Waixenicin A: Exploring Strategies for Nine-Membered Ring Formation.
We present a comprehensive account on our efforts behind the recently published synthesis of waixenicin A. Our approach for constructing the dihydropyran ring relied on an Achmatowicz rearrangement. For the assembly of the nine-membered ring, four distinct strategies were investigated. Our initial attempts using radical-based addition/fragmentation reactions targeting the C7-C11 bond proved unsuitable for accessing the 6/9-bicycle. By employing anionic fragmentation conditions at the furfuryl alcohol stage, we successfully reached a 5/9-bicycle. However, subsequent ring-expansion was unsuccessful. Alternative approaches, such as Nozaki-Hiyama-Kishi or Heck reactions to connect the C6-C7 bond, also encountered difficulties, with no nine-membered ring formation observed. Our first breakthrough came from our attempts to install the C5-C6 bond via an intramolecular alkylation. Surprisingly, subsequent functional group modifications proved unexpectedly challenging, necessitating a redesign of our synthetic route. Drawing from all our investigations, we concluded that construction of the C9-C10 bond would enable efficient nine-membered ring alkylation and would facilitate the installation of the desired substitution pattern along the southern periphery. Exploration of this strategy yielded further surprises but ultimately led to the successful synthesis of waixenicin A and 9-deacetoxy-14,15-deepoxyxeniculin. For the latter compound, a bioinspired one-step rearrangement to xeniafauranol A was achieved.


Transposable gentamicin resistance in IncW plasmids from Hammersmith Hospital.
A transposon, Tn733, encoding the gentamicin acetyltransferase AAC(3) was found on two gentamicin R plasmids of IncW at Hammersmith Hospital. Transposon TN733 has a molecular mass of 5.8 megadaltons and gives a characteristic 2.4 megadalton fragment on digestion with EcoRI. The appearance of gentamicin resistance on a transposon will increase the chances of spread of this gene.


Cellular imaging of zif268 expression in the hippocampus and amygdala during contextual and cued fear memory retrieval: selective activation of hippocampal CA1 neurons during the recall of contextual memories.
The neuroanatomical and molecular basis of fear memory retrieval was studied by analyzing the expression of the plasticity-associated immediate early gene zif268. Cellular quantitative in situ hybridization revealed that zif268 is expressed within specific regions of the hippocampus and amygdala during fear memory retrieval. Within the hippocampus, increased expression of zif268 was observed within CA1 neurons, but not dentate gyrus neurons, during the retrieval of contextual, but not cued, fear associations. In contrast, zif268 expression was increased within neurons of the amygdala (lateral, basal, and central nuclei) during the retrieval of both contextual and cued fear memories. These results demonstrate activation of hippocampal CA1 neurons in contextual fear memory retrieval that was not merely a correlate of the behavioral expression of fear itself, because it was limited to the retrieval of contextual, and not cued, fear memories. Further studies revealed that the selective increase in hippocampal CA1 zif268 expression seen after contextual fear memory retrieval was limited to the retrieval of recent (24 hr) but not older (28 d) memories. These experiments represent the first demonstration that zif268 expression in specific neuronal populations is associated with memory retrieval and suggest that this gene may contribute to plasticity and reconsolidation accompanying the retrieval process.


Central and peripheral effects of CCK receptor antagonists on satiety in dogs.
The role of cholecystokinin (CCK) as a central and peripheral satiety factor was studied using the CCK-B (L-365,260) and CCK-A (MK-329) receptor antagonists in esophageal-fistula dogs. Suppression of feeding was induced by either balloon gastric distension or small bowel nutrient infusion and was measured as the volume sham fed. Intracerebroventricular L-365,260 abolished satiety behavior from gastric distension (volume sham fed 2,667 +/- 211 ml vs. vehicle alone 1,217 +/- 446 ml, P < 0.05) but not small bowel infusion. Intravenous MK-329 abolished satiety behavior from small bowel infusion (volume sham fed 1,900 +/- 521 ml vs. vehicle alone 210 +/- 198 ml, P < 0.05) but not from gastric distension. The volume sham fed after intracerebroventricular MK-329 with balloon gastric distension or small bowel infusion did not differ from control. These results suggest that, in the brain, CCK is a physiological mediator of satiety behavior from gastric distension but not small bowel nutrients and, in the periphery, CCK is a physiological mediator of satiety behavior from small bowel nutrients but not gastric distension.


Current expectations for newborns undergoing the arterial switch operation.
The arterial switch operation (ASO) represents a remarkable success story in the surgical treatment of cyanotic congenital heart disease. This study is designed to assess recent outcomes after the ASO in babies presenting with transposition of the great arteries (TGA) and Taussig-Bing anomaly (TBA). One hundred twenty-five consecutive neonatal and infant ASOs were performed by 2 surgeons at Texas Children's Hospital between July 1, 1995 and October 1, 2003. Patients with TGA and TBA were offered ASO irrespective of patient size and associated cardiac malformations. Primary cardiac diagnoses included TGA with intact ventricular septum (TGA/IVS, n = 79, 63%), TGA with ventricular septal defect (TGA/VSD, n = 37, 30%), and Taussig Bing Anomaly (TBA, n = 9, 7%). With complete follow-up, we observed a 30-day mortality rate of 1.6% (n = 2) with 2 late deaths (1.6%), for an overall actuarial survival rate of 96.3% at 7 years. Although there was a significant incidence of complex coronary ostial origin and branching including single coronary (n = 8, 6.4%) and intramural coronary artery (n = 8, 6.4%), this was not associated with increased operative risk. All patients are fully saturated and NYHA functional class I at latest clinic visit (0.3 to 88.4 months postoperatively). There have been no late coronary events. Of 121 survivors, 7 patients (5.8%) have required cardiovascular reoperation at an average of 15.3 +/-11.7 months postoperatively (range, 3.6 to 30.6 months) for an actuarial freedom from reoperation of 90% at 7 years. Using current methodologies, the ASO can be performed safely and with a low incidence of need for reoperation on intermediate follow-up. Recent experience indicates operative survival rates approaching 100%.


Distributed PACS using distributed file system with hierarchical meta data servers.
In this research, we propose a new distributed PACS (Picture Archiving and Communication Systems) which is available to integrate several PACSs that exist in each medical institution. The conventional PACS controls DICOM file into one data-base. On the other hand, in the proposed system, DICOM file is separated into meta data and image data and those are stored individually. Using this mechanism, since file is not always accessed the entire data, some operations such as finding files, changing titles, and so on can be performed in high-speed. At the same time, as distributed file system is utilized, accessing image files can also achieve high-speed access and high fault tolerant. The introduced system has a more significant point. That is the simplicity to integrate several PACSs. In the proposed system, only the meta data servers are integrated and integrated system can be constructed. This system also has the scalability of file access with along to the number of file numbers and file sizes. On the other hand, because meta-data server is integrated, the meta data server is the weakness of this system. To solve this defect, hieratical meta data servers are introduced. Because of this mechanism, not only fault--tolerant ability is increased but scalability of file access is also increased. To discuss the proposed system, the prototype system using Gfarm was implemented. For evaluating the implemented system, file search operating time of Gfarm and NFS were compared.


Flexibility and Distributive Synthesis Regulate RNA Priming and Handoff in Human DNA Polymerase α-Primase.
DNA replication in eukaryotes relies on the synthesis of a ∼30-nucleotide RNA/DNA primer strand through the dual action of the heterotetrameric polymerase α-primase (pol-prim) enzyme. Synthesis of the 7-10-nucleotide RNA primer is regulated by the C-terminal domain of the primase regulatory subunit (PRIM2C) and is followed by intramolecular handoff of the primer to pol α for extension by ∼20 nucleotides of DNA. Here, we provide evidence that RNA primer synthesis is governed by a combination of the high affinity and flexible linkage of the PRIM2C domain and the surprisingly low affinity of the primase catalytic domain (PRIM1) for substrate. Using a combination of small angle X-ray scattering and electron microscopy, we found significant variability in the organization of PRIM2C and PRIM1 in the absence and presence of substrate, and that the population of structures with both PRIM2C and PRIM1 in a configuration aligned for synthesis is low. Crosslinking was used to visualize the orientation of PRIM2C and PRIM1 when engaged by substrate as observed by electron microscopy. Microscale thermophoresis was used to measure substrate affinities for a series of pol-prim constructs, which showed that the PRIM1 catalytic domain does not bind the template or emergent RNA-primed templates with appreciable affinity. Together, these findings support a model of RNA primer synthesis in which generation of the nascent RNA strand and handoff of the RNA-primed template from primase to polymerase α is mediated by the high degree of inter-domain flexibility of pol-prim, the ready dissociation of PRIM1 from its substrate, and the much higher affinity of the POLA1cat domain of polymerase α for full-length RNA-primed templates.


A novel dysmorphic syndrome with open calvarial sutures and sutural cataracts maps to chromosome 14q13-q21.
We describe a new dysmorphic syndrome in an inbred Saudi Arabian family with 21 members. Five males and one female have similar craniofacial features including wide open calvarial sutures with large and late-closing anterior fontanels, frontal bossing, hyperpigmentation with capillary hemangioma of the forehead, significant hypertelorism, and a broad and prominent nose. In addition, these individuals have Y-shaped sutural cataracts diagnosed by 1-2 years of age. No chromosomal or biochemical abnormalities were identified. A genome-wide scan was performed, and two-point LOD score analysis, assuming autosomal recessive inheritance, detected linkage to chromosome 14q13-q21. The highest LOD scores were obtained for marker GATA136A04 (LOD=4.58 at theta=0.00) and for the adjacent telomeric marker D14S1048 (LOD=4.32 at theta=0.00). Multipoint linkage analysis resulted in a maximum LOD score of 5.44 between markers D14S1048 and GATA136A04. Model independent analysis by SIBPAL confirmed linkage to the same chromosomal region. Haplotype analysis indicated that all affected individuals were homozygous for the interval on chromosome 14q13-q21 with two recombinants for D14S1014 (centromeric) and one recombinant for D14S301 (telomeric). These recombinations limit the disease locus to a region of approximately 7.26 Mb. Candidate genes localized to this region were identified, and analysis of PAX9 did not identify mutations in these patients. The unique clinical phenotype and the mapping data suggest that this family represents a novel autosomal recessive syndrome.


A novel serine-dependent proteolytic activity is responsible for truncated signal transducer and activator of transcription proteins in acute myeloid leukemia blasts.
Hematopoietic cytokine receptor signaling involves activation of signal transducer and activator of transcription (STAT) proteins that are thought to control cellular differentiation. Truncated STAT isoforms (beta forms, rather than the normal alpha forms) have been described and found to block the normal signaling function of the alpha isoforms. We recently demonstrated STATbeta isoforms in bone marrow samples from 21 of 27 (78%) acute myeloid leukemia (AML) patients. We sought to determine the mechanism by which the STATbeta forms were generated. Samples from eight newly diagnosed AML patients were studied; four expressed predominantly STATalpha, and four expressed predominantly STATbeta. The reverse transcription-PCR generated identical products in the two groups, suggesting that alternate mRNA splicing is not responsible for the genesis of STATbeta. Extracts from cells expressing predominantly STATbeta incubated with cell extracts from the MO7E cell line, which expresses predominantly STATa, caused a decrease of the alpha isoforms and an increase of the beta isoforms, suggesting the presence of proteolytic activity. This proteolytic activity was: (a) specific for STAT3 and STAT5, but not for STAT6; (b) serine dependent; (c) equally present in nuclear and cytoplasmic fractions of the leukemic blasts; and (d) different than the activity detected in a murine hematopoietic cell line. The cleaved beta isoforms retained their DNA-binding activity. Because expression of truncated STATs may be involved in blocking differentiation of AML blasts, elucidation of the regulation of the proteolytic activity may contribute to our understanding of leukemogenesis.


Characterization of the immature secretory granule, an intermediate in granule biogenesis.
The events in the biogenesis of secretory granules after the budding of a dense-cored vesicle from the trans-Golgi network (TGN) were investigated in the neuroendocrine cell line PC12, using sulfate-labeled secretogranin II as a marker. The TGN-derived dense-cored vesicles, which we refer to as immature secretory granules, were found to be obligatory organellar intermediates in the biogenesis of the mature secretory granules which accumulate in the cell. Immature secretory granules were converted to mature secretory granules with a half-time of approximately 45 min. This conversion entailed an increase in their size, implying that the maturation of secretory granules includes a fusion event involving immature secretory granules. Pulse-chase labelling of PC12 cells followed by stimulation with high K+, which causes the release of secretogranin II, showed that not only mature, but also immature secretory granules were capable of undergoing regulated exocytosis. The kinetics of secretion of secretogranin II, as well as those of a constitutively secreted heparan sulfate proteoglycan, were reduced by treatment of PC12 cells with nocodazole, suggesting that both secretory granules and constitutive secretory vesicles are transported to the plasma membrane along microtubules. Our results imply that certain membrane proteins, e.g., those involved in the fusion of post-TGN vesicles with the plasma membrane, are sorted upon exit from the TGN, whereas other membrane proteins, e.g., those involved in the interaction of post-TGN vesicles with the cytoskeleton, may not be sorted.


Development of Enzyme Loaded Polyion Complex Vesicle (PICsome): Thermal Stability of Enzyme in PICsome Compartment and Effect of Coencapsulation of Dextran on Enzyme Activity.
Applications of enzymes are intensively studied, particularly for biomedical applications. However, encapsulation or immobilization of enzymes without deactivation and long-term use of enzymes are still at issue. This study focuses on the polymeric vesicles "PICsomes" for encapsulation of enzymes to develop a hecto-nanometer-scaled enzyme-loaded reactor. The catalytic activity of a PICsome-based enzyme nanoreactor is carefully examined to clarify the effect of compartmentalization by PICsome. Encapsulation by PICsome provides a stability enhancement of enzymes after 24 h incubation at 37 °C, which is particularly helpful for maintaining the high effective concentration of β-galactosidase. Moreover, to control the microenvironment inside the nanoreactor, a large amount of dextran, a neutral macromolecule, is encapsulated together with β-galactosidase in the PICsome. The resulting dextran-coloaded nanoreactor contributes to the enhancement of enzyme stability, even after exposure to 24 h incubation at -20 °C, mainly due to the antifreezing effect.


Apoptogenic activity of auraptene of Zanthoxylum schinifolium toward human acute leukemia Jurkat T cells is associated with ER stress-mediated caspase-8 activation that stimulates mitochondria-dependent or -independent caspase cascade.
To isolate pharmacologically safe compounds that can induce apoptosis of tumor cells, leaves of an aromatic plant (Zanthoxylum schinifolium), which are widely used as a food flavor and herbal medicine in Korea and Japan, were sequentially extracted by organic solvents. An apoptogenic ingredient in the methylene chloride extract was further purified by silica gel column chromatography and identified as auraptene (AUR). The IC(50) value of AUR against Jurkat T cells was 16.5 microg/ml. After the treatment of Jurkat T cells with AUR, the endoplasmic reticulum (ER) stress-mediated activation of caspase-12 and -8 and subsequent apoptotic events including c-Jun N-terminal kinase (JNK) activation, cleavage of FLICE inhibitory protein and Bid, mitochondrial cytochrome c release, activation of caspase-9 and -3, degradation of poly (ADP-ribose) polymerase and apoptotic DNA fragmentation were induced in a dose-dependent manner. The cytotoxicity of AUR was not blocked by the anti-Fas neutralizing antibody ZB-4. The AUR-induced cytotoxicity and apoptotic events were abrogated by ectopic over-expression of Bcl-xL or addition of the pan-caspase inhibitor z-VAD-fmk. The individual or simultaneous addition of the m-calpain inhibitor (E64d), JNK inhibitor (SP600125) and mitochondrial permeability transition pore inhibitor (CsA) failed to prevent apoptotic events including caspase-8 activation and Bid cleavage, unless the caspase-8 inhibitor (z-IETD-fmk) was combined, whereas AUR-induced caspase-12 activation was sustained even in the concomitant presence of z-IETD-fmk. These results demonstrated that the apoptotic effect of AUR on Jurkat T cells was exerted by the ER stress-mediated activation of caspase-8, and the subsequent induction of mitochondria-dependent or -independent activation of caspase cascade, which could be suppressed by Bcl-xL.


Engineering β-lactamase zymogens for use in protease activity assays.
In this study, the concept of autoinhibition, a mechanism of protein activity regulation, was applied to the design and engineering of a β-lactamase zymogen. Using this zymogen, a sensitive protease assay method was developed in which activation of the zymogen by proteases produces an amplified absorbance signal. The approach reported here can be adapted for engineering of zymogens as biological sensors and components of synthetic signaling pathways.


[Effects of sigma receptor ligands on psychiatric disorders].
It has recently been suggested that sigma receptors are involved in psychiatric disorders. Sigma 1 receptor antagonists are effective in animal models of positive symptoms, cognitive deficit and disruption of prepulse inhibition in schizophrenia. They also inhibit the development and expression of the conditioned place preference induced by cocaine. On the other hand, sigma 1 receptor agonists reduce the immobility time in the forced swimming and tail suspension tests. Furthermore, sigma 1 receptor agonists attenuate the conditioned fear stress (CFS) response (which is not attenuated by typical anxiolytics or antidepressants) in rodents. The attenuating effects are mediated through sigma 1 receptors, which are closely related to the mesolimbic dopaminergic systems. Sigma 1 receptor agonists also have anti-amnesic effects in various experimental models. Neurosteroids such as dehydroepiandrosterone sulfate and pregnenolone sulfate attenuate the CFS response and have anti-amnesic effects, the effects being mediated via sigma 1 receptors. These findings suggest that sigma receptors are novel potential targets for the treatment of psychiatric disorders such as schizophrenia, drug abuse, depression and dementia.


An interface comprising molecular wires and poly(ethylene glycol) spacer units self-assembled on carbon electrodes for studies of protein electrochemistry.
The characterization and application of a modified electrode interface for protein electrochemistry is reported. This generic interface is composed of a mixed monolayer of oligo(phenylethynylene) molecular wires (MWs) and poly(ethylene glycol) (PEG) deposited on glassy carbon electrodes by reductive adsorption of the respective aryl diazonium salts. Electrochemistry and scanning electron microscopy demonstrate that the PEG component exhibits a distinct decrease in nonspecific adsorption of blood serum and the proteins bovine serum albumin (BSA) and horseradish peroxidase (HRP) relative to a bare glassy carbon electrode. The ability of the MWs to facilitate efficient electron transfer through the PEG layer to the underlying electrode was demonstrated by covalently attaching ferrocenemethylamine to the end of the MWs. The calculated rate constant for this system was 229 +/- 30 s(-1). Covalent attachment of HRP to the MWs allowed direct electron transfer to the redox protein with almost ideal electrochemistry, indicating a specific interaction between the MW and HRP, with a rate constant of 13.4 +/- 2.3 s(-1). This rate constant is more rapid than previously reported for HRP shown to still be catalytically active. Retained catalytic activity of HRP was demonstrated by the enzyme responding to the addition of hydrogen peroxide. Similarly, by attaching myoglobin to the end of the MWs, a rate constant for this protein of 2 s(-1) was measured. The rigidity of the MWs, as well as it being longer than the PEG diluent, means this generic interface can be employed to investigate the electrochemistry of a wide range of redox proteins.


Studies on schistosomiasis. 8. The influence of age on the susceptibility of sheep to infestation with Schistosoma mattheei.
Twenty-eight Dorper wethers, allocated according to age into 4 groups of 7 animals each, and 1 group of 7 Merino wethers, were compared for susceptibility to Schistosoma mattheei infestation. The group mean ages of the Dorper sheep varied from 5-61 months and their live mass from 25-66 kg, while the Merinos were 8 months old and had a mean mass of 19 kg. Despite the marked differences in the age and live mass of the Dorper sheep and the inclusion of 2 breeds in the experiment, no statistically significant differences were found in cercariae which failed to penetrate the sheep the mean percentage development of cercariae to adult worms, worm distribution in the mesenteric and gastric radicles of the portal vein and the pulmonary arterial system, and worms not removed by perfusion. Significant differences between groups (5% significance level) were found, however, in the number of worms recovered from the hepatic portal system, and in the worm sex ratio. On 3 occasions the total number of eggs excreted per female schistosome in the mesentery per 24 hours differed significantly between groups, but each time a different group or groups of sheep were responsible for the variation which was probably due, therefore, not to the age or breed of the sheep, but to daily variations in individuals. Highly significant differences occurred in the infectivity of the 6 cercarial pools used for infestation in spite of standardized collection and handling of the cercariae. Possible reasons for this are discussed and a solution suggested. Frequent egg counts (5 per sheep per week) were done during the first 25 days of patency, until the sheep were slaughtered. Schistosome ova were detected in the faeces of only 1/18 sheep examined on Day +43 after infestation, and 3/17 on Day +44, whereafter this increased rapidly to 15/34 on Day +45, 25/33 on Day +46, etc. A highly significant correlation was found between the total worm egg excretion in the faeces of the sheep per day and the numbers of female schistosomes in the mesentery, especially shortly after the onset of egg production.


The life span of silicone gel breast implants and a comparison of mammography, ultrasonography, and magnetic resonance imaging in detecting implant rupture: a meta-analysis.
Because of the growing concern surrounding the integrity and life span of silicone gel breast implants and the reported variations in the diagnostic accuracy of various imaging techniques in identifying ruptured implants, the authors undertook a meta-analysis of articles in the scientific literature to examine these concerns. They were able to include reports from the literature that detailed the condition and removal of 1,099 breast implants during the past 7 years. The median life span of a silicone gel implant was estimated to be 16.4 years. Of the implants, 79.1% were intact at 10 years, falling to 48.7% by 15 years. The sensitivities and specificities of three imaging modalities used in the diagnosis of implant rupture (mammography, ultrasonography, and magnetic resonance imaging [MRI]) were also evaluated and compared statistically in an effort to discover which of the three techniques might serve as the most reliable screening tool in the diagnosis of gel implant rupture. The sensitivity of mammography for finding a ruptured implant is 28.4% with a specificity of 92.9%. Ultrasonography has a sensitivity and specificity of 59.0% and 76.8% respectively compared with MRI, which was 78.1% and 80.0% respectively. For implants in place for 10 years, one would need to image 3.3 implants by ultrasound to identify a single possible rupture. However, because of the 76.8% specificity, 8.1 implants would need to be imaged to find a confirmed intraoperative rupture. This was similar to MRI, in which 3.1 implants would need to be imaged to detect one suspected rupture, and 6.1 implants would need to be imaged to find one intraoperatively confirmed rupture. The authors do not recommend either ultrasound or MRI as a screening tool based on their meta-analysis.


Non-invasive monitoring of pH and oxygen using miniaturized electrochemical sensors in an animal model of acute hypoxia.
One of the most prevalent causes of fetal hypoxia leading to stillbirth is placental insufficiency. Hemodynamic changes evaluated with Doppler ultrasound have been used as a surrogate marker of fetal hypoxia. However, Doppler evaluation cannot be performed continuously. As a first step, the present work aimed to evaluate the performance of miniaturized electrochemical sensors in the continuous monitoring of oxygen and pH changes in a model of acute hypoxia-acidosis. pH and oxygen electrochemical sensors were evaluated in a ventilatory hypoxia rabbit model. The ventilator hypoxia protocol included 3 differential phases: basal (100% FiO2), the hypoxia-acidosis period (10% FiO2) and recovery (100% FiO2). Sensors were tested in blood tissue (ex vivo sensing) and in muscular tissue (in vivo sensing). pH electrochemical and oxygen sensors were evaluated on the day of insertion (short-term evaluation) and pH electrochemical sensors were also tested after 5 days of insertion (long-term evaluation). pH and oxygen sensing were registered throughout the ventilatory hypoxia protocol (basal, hypoxia-acidosis, and recovery) and were compared with blood gas metabolites results from carotid artery catheterization (obtained with the EPOC blood analyzer). Finally, histological assessment was performed on the sensor insertion site. One-way ANOVA was used for the analysis of the evolution of acid-based metabolites and electrochemical sensor signaling results; a t-test was used for pre- and post-calibration analyses; and chi-square analyses for categorical variables. At the short-term evaluation, both the pH and oxygen electrochemical sensors distinguished the basal and hypoxia-acidosis periods in both the in vivo and ex vivo sensing. However, only the ex vivo sensing detected the recovery period. In the long-term evaluation, the pH electrochemical sensor signal seemed to lose sensibility. Finally, histological assessment revealed no signs of alteration on the day of evaluation (short-term), whereas in the long-term evaluation a sub-acute inflammatory reaction adjacent to the implantation site was detected. Miniaturized electrochemical sensors represent a new generation of tools for the continuous monitoring of hypoxia-acidosis, which is especially indicated in high-risk pregnancies. Further studies including more tissue-compatible material would be required in order to improve long-term electrochemical sensing.


Surgical management of conjunctival tumors. The 1994 Lynn B. McMahan Lecture.
To our knowledge, there are no articles that describe the specific step-by-step details of the surgical removal of premalignant and malignant conjunctival tumors. We describe our current approach to the surgical management of squamous cell carcinoma (intraepithelial or invasive), localized melanoma, and primary acquired melanosis of the conjunctiva. The surgical method differs with limbal tumors, extralimbal tumors, and primary acquired melanosis. Limbal lesions are managed by localized alcohol corneal epitheliectomy, removal of the main mass by a partial lamellar scleroconjunctivectomy, and supplemental cryotherapy. Tumors located in the extralimbal conjunctiva are managed by alcohol application, wide circumferential surgical resection, and cryotherapy. Primary acquired melanosis is managed by alcohol epitheliectomy, removal of suspicious foci, quadrantic staging biopsies, and cryotherapy from the underside of the conjunctiva. In all cases, a "no touch" method is used and direct manipulation of the tumor is avoided to prevent tumor cell seeding into a new area. We have employed this technique on 109 patients with conjunctival squamous neoplasms and 137 patients with conjunctival melanoma, about 80 of which neoplasms were associated with primary acquired melanosis. Our observations suggest that well-planned initial surgical management using this technique decreases the chance of tumor recurrence for conjunctival melanoma and squamous cell carcinoma. We describe a detailed stepwise approach to the surgical management of conjunctival neoplasms. It requires meticulous clinical evaluation and complete removal of the tumor in one operation using a specific technique.


Effect of Parecoxib Sodium on Myocardial Ischemia-Reperfusion Injury Rats.
BACKGROUND We aimed to explore the effect of parecoxib sodium on myocardial ischemia-reperfusion (I/R) injury rats and its mechanism. MATERIAL AND METHODS The coronary artery of Sprague-Dawley rats was occluded for 6 h of myocardial ischemia, followed by reperfusion for 30 min (I/R group). Before ischemia, parecoxib sodium (10 mg/kg) was intraperitoneally injected twice a day for 3 consecutive days, followed by reperfusion for 6 h (I/R+Pare group). The cardiac function and changes in the infarction area were evaluated via echocardiography in each group. The differences in the expressions of apoptosis-related proteins were determined via immunohistochemistry and western blotting. Then, the percentage of reactive oxygen species (ROS)⁺ cells and the content of lipid peroxide were detected, based on which the degree of oxidative stress was evaluated. Next, the expressions of nuclear factor-kappaB (NF-kappaB) and nuclear factor E2-related factor 2 (Nrf-2) signaling pathways and downstream target genes were determined using real-time quantitative polymerase chain reaction (PCR). RESULTS After treatment with parecoxib sodium, the cardiac function of I/R injury rats was restored, and the infarction area and apoptosis level were reduced (P<0.05). Parecoxib sodium reduced the levels of ROS and lipid peroxidation in myocardial I/R injury rats, thereby weakening oxidative stress. It also regulated the redox imbalance caused by I/R injury through regulating NF-kappaB and Nrf-2 (P<0.01). In addition, after treatment with parecoxib sodium, NF-kappaB was significantly downregulated, while Nrf-2 was upregulated, and the content of proinflammatory cytokines was obviously reduced (P<0.01). CONCLUSIONS Parecoxib sodium exerts a protective effect against myocardial I/R injury through regulating antioxidant and inflammatory mechanisms.


Quality improvement implementation and disparities: the case of the health disparities collaboratives.
The Health Disparities Collaboratives (HDCs), a quality improvement (QI) collaborative incorporating rapid QI, a chronic care model, and learning sessions, have been implemented in over 900 community health centers across the country. To determine the HDC's effect on clinical processes and outcomes, their financial impact, and factors important for successful implementation. Systematic review of the literature. The HDCs improve clinical processes of care over short-term period of 1 to 2 years, and clinical processes and outcomes over longer period of 2 to 4 years. Most participants perceive that the HDCs are successful and worth the effort. Analysis of the Diabetes Collaborative reveals that it is societally cost-effective, with an incremental cost-effectiveness ratio of $33,386 per quality-adjusted life year, but that consistent revenue streams for the initiative do not exist. Common barriers to improvement include lack of resources, time, and staff burnout. Highest ranked priorities for more funding are money for direct patient services, data entry, and staff time for QI. Other common requests for more assistance are help with patient self-management, information systems, and getting providers to follow guidelines. Relatively low-cost ways to increase staff morale and prevent burnout include personal recognition, skills development opportunities, and fair distribution of work. The HDCs have successfully improved quality of care, and the Diabetes Collaborative is societally cost-effective, but policy reforms are necessary to create a sustainable business case for these health centers that serve many uninsured and underinsured populations.


The sequential catheterization amid progressive endograft deployment technique for fenestrated endovascular aortic aneurysm repair.
Fenestrated endovascular aneurysm repair (FEVAR) is an alternative to open repair of complex aortic aneurysms. Despite promising short-term results, the technical complexities of this procedure remain a considerable challenge. The risk of technical failure with loss of visceral or renal arteries is ubiquitous even in the most experienced hands, and thus many patients with unfavorable anatomy are frequently denied FEVAR. We have adopted a new technique for FEVAR that involves retrograde brachial artery access and stepwise deployment of the endograft during target vessel catheterization, overcoming many anatomic limitations encountered from a transfemoral approach. This technique, termed sequential catheterization amid progressive endograft deployment, has become our preferred approach for FEVAR and is described in this article. Of note, currently available Food and Drug Administration-approved fenestrated endografts may not be amenable to sequential catheterization amid progressive endograft deployment as this technique requires preloaded wires incorporated into the endografts.


[Determination of the carbamate kinase activity of bacteria].
The activity of carbamate kinase (EC 18.104.22.168) was determined in bacteria using a simple modified procedure. Carbamoyl phosphate produced under the action of carbamate kinase carbamoylated ammonia in a reaction which was not enzyme-catalyzed yielding urea that was assayed by the colorimetric technique. The activity of carbamate kinase was found by this method in a number of microorganisms. The method can be used to study other enzymes synthesizing carbamoyl phosphate. The advantages of the method over other techniques are discussed.


Intermediate endocrine-acinar pancreatic cells in duct ligation conditions.
When tissues were subjected to 24 h of duct ligation, intermediate pancreatic cells simultaneously displaying endocrine and exocrine phenotypes appeared. Immunocytochemistry by laser scanning confocal microscopy revealed the appearance of a large number of these cells coexpressing insulin and amylase. These cells were located within the islets of Langerhans as well as in the acinar parenchyma. They were also detected in a culture system of isolated pancreatic cells. With the use of immunoelectron microscopy, two types of secretory granules were identified in these cells. One was insulin immunoreactive, whereas the other, resembling zymogen granules, contained amylase. Occasionally, some small granules displayed a double labeling for both secretory proteins. Numerous crinophagic bodies and autophagosomes containing insulin and/or amylase were also present. In situ hybridization, applied with the specific probes, confirmed the presence of both insulin and amylase mRNAs in these cells. Because duct ligation is known to induce insulin cell proliferation, the present results confirm that endocrine-acinar cells do appear in such condition and may represent intermediate steps in a transdifferentiating process.


The importance of rectal cancer MRI protocols on interpretation accuracy.
Magnetic resonance imaging (MRI) is used for preoperative local staging in patients with rectal cancer. Our aim was to retrospectively study the effects of the imaging protocol on the staging accuracy. MR-examinations of 37 patients with locally advanced disease were divided into two groups; compliant and noncompliant, based on the imaging protocol, without knowledge of the histopathological results. A compliant rectal cancer imaging protocol was defined as including T2-weighted imaging in the sagittal and axial planes with supplementary coronal in low rectal tumors, alongside a high-resolution plane perpendicular to the rectum at the level of the primary tumor. Protocols not complying with these criteria were defined as noncompliant. Histopathological results were used as gold standard. Compliant rectal imaging protocols showed significantly better correlation with histopathological results regarding assessment of anterior organ involvement (sensitivity and specificity rates in compliant group were 86% and 94%, respectively vs. 50% and 33% in the noncompliant group). Compliant imaging protocols also used statistically significantly smaller voxel sizes and fewer number of MR sequences than the noncompliant protocols Appropriate MR imaging protocols enable more accurate local staging of locally advanced rectal tumors with less number of sequences and without intravenous gadolinium contrast agents.


Therapeutic options in hypertrophic cardiomyopathy: a pediatric perspective.
Hypertrophic cardiomyopathy (HCM) is the most prevalent genetic cardiomyopathy in children and young adults. Despite advances in understanding the molecular etiology of this disorder, the clinical phenotypes vary widely from asymptomatic septal hypertrophy, to frank congestive heart failure due to left ventricular outflow tract obstruction (LVOTO), to unexpected sudden cardiac death. Thus, isolating a specific genetic defect in this disease does little to predict the clinical consequence. This is best seen in large families with several people affected by HCM who all have the identical mutation but who may have very different clinical phenotypes. Evaluating individuals with HCM consists of establishing the diagnosis through cardiac imaging, noninvasively assessing the risk for sudden death, and medically or surgically treating symptomatic LVOTO when it is present.


Reliability, Sensitivity, and Minimal Detectable Change of a New Specific Climbing Test for Assessing Asymmetry in Reach Technique.
Čular, D, Dhahbi, W, Kolak, I, Iacono, AD, Bešlija, T, Laffaye, G, and Padulo, J. Reliability, sensitivity, and minimal detectable change of a new specific climbing test for assessing asymmetry in reach technique. J Strength Cond Res 35(2): 527-534, 2021-The aims of this study were to establish intertrial and intersession reliability, sensitivity, and minimal detectable change of a new climbing test specifically for assessing asymmetry in reach technique (TEST). Twenty-four young climbers (16 males and 8 females) participated in this study. The protocol consisted of performing, in counterbalanced random order, 3 tests; TEST, maximum handgrip force, and squat on the bench, in 2 sessions (with 3 trials for each session). TEST performance was expressed as: TEST performance for left hand (TESTL), TEST performance for right hand (TESTR), and absolute symmetry index (ASI). For intertrial and intersession reliability assessment, TESTL and TESTR showed excellent reliability (intraclass correlation coefficients ranged: 0.96-1.00; SEM% ranged: 0.07-1.23; and coefficient of variation-CV%: 1.28-2.53). In addition, SEMs were smaller than the smallest worthwhile change (SWC) values (SWC% = 1.07 and 0.99 for TESTL and TESTR, respectively), and the minimal detectable change (MDC95) for both sides was small (<4.36 cm). An exception was ASI, which showed low absolute reliability and marginal sensitivity (SEM% = 15.13 > SWC% = 8.40 and CV% = 41.98). Pairwise test comparisons revealed no difference between sides. Considering the high reliability and the satisfactory sensitivity, TEST can be used to define individual asymmetry in the performance of the reach technique to the left or the right body side in climbers. However, interpreting data using the ASI index requires caution because it had poor absolute reliability and marginal sensitivity.


The Effect of an Educational and Interactive Informed Consent Process on Patients With Cervical Spondylotic Myelopathy Caused by Ossification of the Posterior Longitudinal Ligament.
Prospective observational cohort study. In this study, an educational and interactive informed consent (EIC) program was proposed for patients with cervical spondylotic myelopathy caused by ossification of the posterior longitudinal ligament (OPLL-CSM) to improve their comprehension level during the informed consent process. OPLL-CSM is a slow progressive disease, and it is difficult for patients to understand the disease. Few studies have evaluated very specific programs to improve the informed consent process for these patients. This prospective study evaluated patients with OPLL-CSM who either underwent the proposed EIC process (n = 63) or the standard consent process (n = 124). The standard consent process only included a physician-patient interview. During the EIC process, information was provided regarding OPLL-CSM through information booklets, a video, verbal information, and initial and second physician-patient interviews. After the second physician-patient interview, the patient was requested to answer 14 medical questions to assess their knowledge about OPLL-CSM. The proposed EIC process took approximately 90 minutes. They were asked to report the most useful educational method and the most effective method of reinforcing verbal communication. The mean questionnaire scores were higher in the EIC group than in the control group (P < 0.001). Video was selected by 50/63 patients (79.4%) as the most useful EIC process method, and the most effective method of reinforcing verbal communication was video (n = 61; 96.8%). Patients in the EIC group reported having higher satisfaction with surgery (P = 0.024) than did those in the control group. The proposed EIC process was shown to result in good patient comprehension and recall regarding OPLL-CSM. Using a video was the most informative and effective reinforcement of verbal communication. The enhanced educational group had better knowledge and improved satisfaction following surgery. The EIC process might help physicians educate and counsel patients regarding OPLL-CSM and its treatment. 2.


Histone variants at the nexus of eukaryotic chromatin readouts, stress and plant development.
Histones are crucial proteins that are involved in packaging the DNA as condensed chromatin inside the eukaryotic cell nucleus. Rather than being static packaging units, these molecules undergo drastic variations spatially and temporally to facilitate accessibility of DNA to replication, transcription as well as wide range of gene regulatory machineries. In addition, incorporation of paralogous variants of canonical histones in the chromatin is ascribed to specific functions. Given the peculiar requirement of plants to rapidly modulate gene expression levels on account of their sessile nature, histones and their variants serve as additional layers of gene regulation. This review summarizes the mechanisms and implications of distribution, modifications and differential incorporation of histones and their variants across plant genomes and outlines emerging themes.


Preliminary results for avelumab plus axitinib as first-line therapy in patients with advanced clear-cell renal-cell carcinoma (JAVELIN Renal 100): an open-label, dose-finding and dose-expansion, phase 1b trial.
The combination of an immune checkpoint inhibitor and a VEGF pathway inhibitor to treat patients with advanced renal-cell carcinoma might increase the clinical benefit of these drugs compared with their use alone. Here, we report preliminary results for the combination of avelumab, an IgG1 monoclonal antibody against the programmed cell death protein ligand PD-L1, and axitinib, a VEGF receptor inhibitor approved for second-line treatment of advanced renal-cell carcinoma, in treatment-naive patients with advanced renal-cell carcinoma. The JAVELIN Renal 100 study is an ongoing open-label, multicentre, dose-finding, and dose-expansion, phase 1b study, done in 14 centres in the USA, UK, and Japan. Eligible patients were aged 18 years or older (≥20 years in Japan) and had histologically or cytologically confirmed advanced renal-cell carcinoma with clear-cell component, life expectancy of at least 3 months, an Eastern Cooperative Oncology Group performance status of 1 or less, received no previous systemic treatment for advanced renal cell carcinoma, and had a resected primary tumour. Patients enrolled into the dose-finding phase received 5 mg axitinib orally twice daily for 7 days, followed by combination therapy with 10 mg/kg avelumab intravenously every 2 weeks and 5 mg axitinib orally twice daily. Based on the pharmacokinetic data from the dose-finding phase, ten additional patients were enrolled into the dose-expansion phase and assigned to this regimen. The other patients in the dose-expansion phase started taking combination therapy directly. The primary endpoint was dose-limiting toxicities in the first 4 weeks (two cycles) of treatment with avelumab plus axitinib. Safety and antitumour activity analyses were done in all patients who received at least one dose of avelumab or axitinib. This trial is registered with ClinicalTrials.gov, number NCT02493751. Between Oct 30, 2015, and Sept 30, 2016, we enrolled six patients into the dose-finding phase and 49 into the dose-expansion phase of the study. One dose-limiting toxicity of grade 3 proteinuria due to axitinib was reported among the six patients treated during the dose-finding phase. At the cutoff date (April 13, 2017), six (100%, 95% CI 54-100) of six patients in the dose-finding phase and 26 (53%, 38-68) of 49 patients in the dose-expansion phase had confirmed objective responses (32 [58%, 44-71] of all 55 patients). 32 (58%) of 55 patients had grade 3 or worse treatment-related adverse events, the most frequent being hypertension in 16 (29%) patients and increased concentrations of alanine aminotransferase, amylase, and lipase, and palmar-plantar erythrodysaesthesia syndrome in four (7%) patients each. Six (11%) of 55 patients died before data cutoff, five (9%) due to disease progression and one (2%) due to treatment-related autoimmune myocarditis. At the end of the dose-finding phase, the maximum tolerated dose established for the combination was avelumab 10 mg/kg every 2 weeks and axitinib 5 mg twice daily. The safety profile of the combination avelumab plus axitinib in treatment-naive patients with advanced renal-cell carcinoma seemed to be manageable and consistent with that of each drug alone, and the preliminary data on antitumour activity are encouraging. A phase 3 trial is assessing avelumab and axitinib compared with sunitinib monotherapy. Pfizer and Merck.


Evolutionarily divergent herpesviruses modulate T cell activation by targeting the herpesvirus entry mediator cosignaling pathway.
The herpesvirus entry mediator (HVEM), a member of the TNF receptor (TNFR) superfamily, can act as a molecular switch that modulates T cell activation by propagating positive signals from the TNF-related ligand LIGHT (TNFR superfamily 14), or inhibitory signals through the Ig superfamily member B and T lymphocyte attenuator (BTLA). Competitive binding analysis and mutagenesis reveals a unique BTLA binding site centered on a critical lysine residue in cysteine-rich domain 1 of HVEM. The BTLA binding site on HVEM overlaps with the binding site for the herpes simplex virus 1 envelope glycoprotein D, but is distinct from where LIGHT binds, yet glycoprotein D inhibits the binding of both ligands, potentially nullifying the pathway. The binding site on HVEM for BTLA is conserved in the orphan TNFR, UL144, present in human CMV. UL144 binds BTLA, but not LIGHT, and inhibits T cell proliferation, selectively mimicking the inhibitory cosignaling function of HVEM. The demonstration that distinct herpesviruses target the HVEM-BTLA cosignaling pathway suggests the importance of this pathway in regulating T cell activation during host defenses.


Kindlin-1 regulates integrin dynamics and adhesion turnover.
Loss-of-function mutations in the gene encoding the integrin co-activator kindlin-1 cause Kindler syndrome. We report a novel kindlin-1-deficient keratinocyte cell line derived from a Kindler syndrome patient. Despite the expression of kindlin-2, the patient's cells display several hallmarks related to reduced function of β1 integrins, including abnormal cell morphology, cell adhesion, cell spreading, focal adhesion assembly, and cell migration. Defective cell adhesion was aggravated by kindlin-2 depletion, indicating that kindlin-2 can compensate to a certain extent for the loss of kindlin-1. Intriguingly, β1 at the cell-surface was aberrantly glycosylated in the patient's cells, and its expression was considerably reduced, both in cells in vitro and in the patient's epidermis. Reconstitution with wild-type kindlin-1 but not with a β1-binding defective mutant restored the aberrant β1 expression and glycosylation, and normalized cell morphology, adhesion, spreading, and migration. Furthermore, the expression of wild-type kindlin-1, but not of the integrin-binding-defective mutant, increased the stability of integrin-mediated cell-matrix adhesions and enhanced the redistribution of internalized integrins to the cell surface. Thus, these data uncover a role for kindlin-1 in the regulation of integrin trafficking and adhesion turnover.


Comparing Utility Scores in Common Spinal Radiculopathies: Results of a Prospective Valuation Study.
Study Design Prospective observational study. Objective To determine whether preference-based health utility scores for common spinal radiculopathies vary by specific spinal level. Methods We employed a standard gamble study using the general public to calculate individual preference-based quality of life for four common radiculopathies: C6, C7, L5, and S1. We compared utility scores obtained for each level of radiculopathy with analysis of variance and t test. Multivariable regression was used to test the effects of the covariates age, sex, and years of education. We also reviewed the literature for publications reporting EuroQol-5 Dimensions (EQ-5D) scores for patients with radiculopathy. Results Two hundred participants were included in the study. Average utility for the four spinal levels fell within a narrow range (0.748 to 0.796). There were no statistically significant differences between lumbar and cervical radiculopathies, nor were there significant differences among the different spinal levels (F = 0.0850, p = 0.086). Age and sex had no significant effect on utility scores. There was a significant correlation between years of education and utility values for S1 radiculopathy (p = 0.037). On review of the literature, no study separated utility values by specific spinal level. EQ-5D utilities for both cervical and lumbar radiculopathy were considerably lower than the results of our study. Conclusions Utility values associated with the most common levels of cervical and lumbar radiculopathy do not significantly differ from each other, validating the current practice of grouping utility by spinal segment rather than by specific root levels. The discrepancy in average utility values between our study and the EQ-5D highlights the need to be mindful of the underlying instruments used when assessing outcomes studies from different sources.


Identification of the advanced glycation end products N(epsilon)-carboxymethyllysine in the synovial tissue of patients with rheumatoid arthritis.
Generation of advanced glycation end products (AGEs) is an inevitable process in vivo and can be accelerated under pathological conditions such as oxidative stress. In serum and synovial fluid of patients with rheumatoid arthritis (RA) raised AGE levels have been found. To determine the presence of N(epsilon)-carboxymethyllysine (CML; marker of oxidative stress) in RA synovial tissue by immunohistology. Frozen synovial tissue samples from 10 patients with RA and eight controls (four patients without joint disease and four patients with osteoarthritis (OA)) were treated with rabbit-anti-CML-IgG and goat-antirabbit-IgG. Immunostaining was visualised by streptavidine-alkaline phosphatase (chromogen fuchsin). Cell differentiation was performed with antibodies against CD68, CD45RO, and CD20. CML was detected in the synovial lining, sublining, and endothelium in 10/10 RA and 4/4 OA synovial specimens. In RA some macrophages (CD68+) and T cells (CD45RO+) showed positive immunostaining for CML, whereas B cells were negative. Staining in OA synovial sublining was weak compared with RA. CML was detected for the first time in RA and OA synovial tissue. Different patterns of immunostaining in RA and OA and the presence of CML on macrophages and T cells, suggest a role for CML in the pathogenesis of RA. This might be due to presentation of new epitopes which can maintain or even trigger an autoimmune response.


Long term effects of high intensity laser therapy in lateral epicondylitis patients.
The objective of this study is to investigate short- and long-term effects of high-intensity laser therapy (HILT) in lateral epicondylitis (LE) patients. Thirty patients with LE diagnosis (23 unilateral and 7 bilateral in total 37 elbows) were treated using HILT. LE patients were evaluated before, right after, and 6 months following HILT intervention post-treatment using visual analogue scale for pain (VAS) during activity and resting. Disabilities of the Arm, Shoulder, and Hand (DASH) Score and hand grip strength test (HGST) were used. The participants of the present study were also evaluated using Short-Form 36 (SF-36) before and 6 months after the treatment. Out of the 30 patients, 8 were male and 22 female with a mean age of 47.2 ± 9.7. The activity and resting VAS, DASH, and HGST scores revealed statistically significant improvement (p = 0.001) following treatment. Whereas VAS activity, DASH, and HGST scores increased after treatment until post-treatment 6 months significantly (p = 0.001), VAS resting scores remained stable (p = 0.476). A statistically significant improvement was also evident in the physical and mental components of SF-36 scores following treatment until post-treatment 6 months compared to pre-treatment scores (p = 0.001). In conclusion, the results of the present study suggest that HILT is a reliable, safe, and effective treatment option in LE patients in the short and long term considering pain, functional status, and quality of life.


The influence of sodium on liking and consumption of salty food.
Excessive sodium (Na) intake has been linked to development of hypertension and related pathologies. In this study, we assessed if the sodium chloride (NaCl) concentration in a prototypical food influences the liking and intake of that food. In study 1, detection and recognition thresholds for NaCl were assessed, and perceived salt intensity and liking for hash browns of varying sodium concentrations (40 mg, 120 mg, 170 mg, and 220 mg Na/100 g) were compared in a lab setting. In study 2, detection and recognition thresholds for NaCl were assessed in a lab setting, and lunches consisting of hash browns, basic salad, and beverages were consumed freely in a dining setting on 4 separate occasions. Intake and liking ratings for hash browns were recorded after the lunch. In both studies, detection and recognition thresholds for NaCl were not associated with perceived saltiness, liking, or intake of hash browns. Liking and perceived salt taste intensity of hash browns were correlated (r = 0.547 P < 0.01), and in study 1 the 220 mg sodium hash brown was most liked (P < 0.05). In study 2, there was no association between Na concentration and liking or consumption of hash browns. In summary, liking of hash browns were influenced by whether testing was in a lab or dining room environment. In a dining room environment, large decreases (>50%) of sodium content of food were achievable with only minor decrease in liking and no effect on consumption of the food.


Effects of fatty (fa) allele and high-fat diet on adipose tissue leptin and lipid metabolism.
Leptin is an adipocyte-secreted hormone that binds hypothalamic receptors and potently decreases food intake. Leptin receptor defects in homozygous mutant Zucker fatty ( fa/fa) rats lead to massive obesity, hyperphagia, decreased energy expenditure, and insulin resistance, while the phenotype of heterozygous ( Fa/fa) lean rats lies between lean ( Fa/Fa) and obese ( fa/fa) rats. Whether heterezygotes exhibit specific changes in lipid metabolism in a diet-responsive manner is not clear. Thus, the specific aim of this study was to test whether the presence of one fa allele modulates lipid metabolism and leptin, and whether these effects are exacerbated by high-fat diet. We demonstrate that the presence of one fa allele significantly increases lipogenesis in adipose tissue assessed by glycerol-3-phosphate dehydrogenase (GPDH) and fatty acid synthase (FAS) activities. FAS is more responsive to high-fat diets than GPDH in Fa/fa rats. Adipose tissue leptin levels are significantly higher in fat pads of Fa/fa compared to Fa/Fa rats. Moreover, Fa/fa rats fed high-fat diet show an additional two-fold increase in leptin levels compared to wild type rats on the same diet. Collectively, these results indicate that the presence of one fa allele increase adipocyte lipogenic enzyme activities, which results in hyperleptinemia concurrent with increased adiposity.


The effects of forestry on Hg bioaccumulation in nemoral/boreal waters and recommendations for good silvicultural practice.
Mercury (Hg) levels are alarmingly high in fish from lakes across Fennoscandia and northern North America. The few published studies on the ways in which silviculture practices influence this problem indicate that forest operations increase Hg in downstream aquatic ecosystems. From these studies, we estimate that between one-tenth and one-quarter of the Hg in the fish of high-latitude, managed forest landscapes can be attributed to harvesting. Forestry, however, did not create the elevated Hg levels in the soils, and waterborne Hg/MeHg concentrations downstream from harvested areas are similar to those from wetlands. Given the current understanding of the way in which silviculture impacts Hg cycling, most of the recommendations for good forest practice in Sweden appear to be appropriate for high-latitude regions, e.g., leaving riparian buffer zones, as well as reducing disturbance at stream crossings and in moist areas. The recommendation to restore wetlands and reduce drainage, however, will likely increase Hg/MeHg loadings to aquatic ecosystems.


Fine-needle aspiration biopsy of adrenal masses in patients with extraadrenal malignancy.
Reports evaluating the efficacy of fine needle aspiration (FNA) of the adrenal gland have suggested a possible correlation between size of an adrenal mass and the presence of a primary or metastatic malignancy. These studies have focused on FNAs of all adrenal gland masses regardless of clinical history. This study investigates this relationship in a subpopulation of patients with a known history of primary extraadrenal malignancy. All patients who have undergone computed tomographic-guided FNA biopsy of an adrenal mass at Loyola University Medical Center and Hines Veterans Administration Hospital, from 1985 to 1991, were reviewed. If size was assumed to be an independent predictor for presence of metastases, the highest efficiency was obtained with a cutoff value of 3 cm. This value divided the group into 15 (42%) "low-risk" (< or = 3 cm) and 21 (58%) "high-risk" (> 3 cm) subjects. In the "low-risk" group, 87% of the masses (13 of 15) were benign and 13% (2 of 15) were malignant. Within the "high-risk" group, more than 95% of the masses (20 of 21) were malignant, with a single (5%) benign case (p < 0.05). There is a significant correlation between the size of an adrenal nodule and the presence of metastases in patients with a known primary extraadrenal malignancy. Nodules greater than 3 cm have a very high probability of involvement by metastatic tumor. Nodules 3 cm or smaller are usually benign, but metastatic tumor can still be found in up to 13%. FNA biopsy is useful in evaluating adrenal masses in this setting.


Methylphenidate does not influence smoking-reinforced responding or attentional performance in adult smokers with and without attention deficit hyperactivity disorder (ADHD).
Individuals with Attention Deficit Hyperactivity Disorder (ADHD) smoke cigarettes at rates higher than the general population and questions have been raised about how stimulant drugs-the frontline pharmacological treatment for ADHD-influence smoking risk and behavior in those with ADHD. In the present study adult regular smokers with (n = 16) and without (n = 17) ADHD participated in 3 experimental sessions in which they completed a Progressive Ratio (PR) task to measure the relative reinforcing effects of cigarette smoking and money after oral administration of placebo and 2 active doses of methylphenidate (10 mg and 40 mg). We also measured attention and inhibitory control via a Continuous Performance Test (CPT). Methylphenidate had no effect on smoking-reinforced responding, attention, or inhibitory control in either group. Attention and inhibitory control were associated with smoking-reinforced responding, but unsystematically and only in the non-ADHD group. Several design features, such as the value of the monetary response option, the PR schedule, and the potential effects of smoking on attention and inhibitory control, could have contributed to the negative findings and are discussed as such. Although inconsistent with some previous human laboratory studies of stimulant drugs and smoking, results are consistent with recent trials of stimulant drugs as adjuncts for smoking cessation in adult smokers with ADHD. In general, methylphenidate at mild and moderate doses did not influence the relative reinforcing effects of cigarette smoking in adults with and without ADHD.


Structure-activity relationships of sulfonamides derived from carvacrol and their potential for the treatment of Alzheimer's disease.
Five synthetic sulfonamides derived from carvacrol, a natural product and a small molecule with druglike properties, were evaluated with respect to their effects on the cognitive deficits of animals with streptozotocin (STZ)-induced Alzheimer's disease (AD). Memory, ambulation, anxiety and oxidative stress were evaluated. In vitro assays were performed to assess the inhibition of acetylcholinesterase (AChE), and the data were combined with molecular docking for the establishment of structure-activity relationships. The memories of animals treated with the compounds derived from morpholine (1), hydrazine (3) and 2-phenol (5) were improved. Compound 3 was the most promising, yielding excellent results in the inhibitory avoidance test. Moreover, the compounds did not exhibit any deleterious effects on the animals' ambulation in the open field test. Molecular docking confirmed the results obtained in the AChE inhibition assay. In short, compounds 1, 3 and 5 can reduce STZ-induced deficits and show potential for the treatment of Alzheimer's. In addition, these agents produce significant anxiolytic and antioxidant effects.


Characteristics of 698 patients with dissociative seizures: A UK multicenter study.
We aimed to characterize the demographics of adults with dissociative (nonepileptic) seizures, placing emphasis on distribution of age at onset, male:female ratio, levels of deprivation, and dissociative seizure semiology. We collected demographic and clinical data from 698 adults with dissociative seizures recruited to the screening phase of the CODES (Cognitive Behavioural Therapy vs Standardised Medical Care for Adults With Dissociative Non-Epileptic Seizures) trial from 27 neurology/specialist epilepsy clinics in the UK. We described the cohort in terms of age, age at onset of dissociative seizures, duration of seizure disorder, level of socioeconomic deprivation, and other social and clinical demographic characteristics and their associations. In what is, to date, the largest study of adults with dissociative seizures, the overall modal age at dissociative seizure onset was 19 years; median age at onset was 28 years. Although 74% of the sample was female, importantly the male:female ratio varied with age at onset, with 77% of female but only 59% of male participants developing dissociative seizures by the age of 40 years. The frequency of self-reported previous epilepsy was 27%; nearly half of these epilepsy diagnoses were retrospectively considered erroneous by clinicians. Patients with predominantly hyperkinetic dissociative seizures had a shorter disorder duration prior to diagnosis in this study than patients with hypokinetic seizures (P < .001); dissociative seizure type was not associated with gender. Predominantly hyperkinetic seizures were most commonly seen in patients with symptom onset in their late teens. Thirty percent of the sample reported taking antiepileptic drugs; this was more common in men. More than 50% of the sample lived in areas characterized by the highest levels of deprivation, and more than two-thirds were unemployed. Females with dissociative seizures were more common at all ages, whereas the proportion of males increased with age at onset. This disorder was associated with socioeconomic deprivation. Those with hypokinetic dissociative seizures may be at risk for delayed diagnosis and treatment.


Quantification of volumetric and geometric changes occurring during fractionated radiotherapy for head-and-neck cancer using an integrated CT/linear accelerator system.
Many patients receiving fractionated radiotherapy (RT) for head-and-neck cancer have marked anatomic changes during their course of treatment, including shrinking of the primary tumor or nodal masses, resolving postoperative changes/edema, and changes in overall body habitus/weight loss. We conducted a pilot study to quantify the magnitude of these anatomic changes with systematic CT imaging. Fourteen assessable patients were enrolled in this pilot study. Eligible patients had to have a pathologic diagnosis of head-and-neck cancer, be treated with definitive external beam RT, and had have gross primary and/or cervical nodal disease measuring at least 4 cm in maximal diameter. All patients were treated using a new commercial integrated CT-linear accelerator system (EXaCT) that allows CT imaging at the daily RT sessions while the patient remains immobilized in the treatment position. CT scans were acquired three times weekly during the entire course of RT, and both gross tumor volumes (GTVs: primary tumor and involved lymph nodes) and normal tissues (parotid glands, spinal canal, mandible, and external contour) were manually contoured on every axial slice. Volumetric and positional changes relative to a central bony reference (the center of mass of the C2 vertebral body) were determined for each structure. Gross tumor volumes decreased throughout the course of fractionated RT, at a median rate of 0.2 cm(3) per treatment day (range, 0.01-1.95 cm(3)/d). In terms of the percentage of the initial volume, the GTVs decreased at a median rate of 1.8%/treatment day (range, 0.2-3.1%/d). On the last day of treatment, this corresponded to a median total relative loss of 69.5% of the initial GTV (range, 9.9-91.9%). In addition, the center of the mass of shrinking tumors changed position with time, indicating that GTV loss was frequently asymmetric. At treatment completion, the median center of the mass displacement (after corrections for daily setup variation) was 3.3 mm (range, 0-17.3 mm). Parotid glands also decreased in volume (median, 0.19 cm(3)/d range, 0.04-0.84 cm(3)/d), and generally shifted medially (median, 3.1 mm; range, 0-9.9 mm) with time. This medial displacement of the parotid glands correlated highly with the weight loss that occurred during treatment. Measurable anatomic changes occurred throughout fractionated external beam RT for head-and-neck cancers. These changes in the external contour, shape, and location of the target and critical structures appeared to be significant during the second half of treatment (after 3-4 weeks of treatment) and could have potential dosimetric impact when highly conformal treatment techniques are used. These data may, therefore, be useful in the development of an adaptive RT scheme (periodic adjustment of the conformal treatment plan) that takes into account such treatment-related anatomic changes. In theory, such a strategy would maximize the therapeutic ratio of RT.


Absence of complementary sex determination in two Leptopilina species (Figitidae, Hymenoptera) and a reconsideration of its incompatibility with endosymbiont-induced thelytoky.
Complementary sex determination (CSD) is a widespread sex determination mechanism in haplodiploid Hymenoptera. Under CSD, sex is determined by the allelic state of one or multiple CSD loci. Heterozygosity at one or more loci leads to female development, whereas hemizygosity of haploid eggs and homozygosity of diploid eggs results in male development. Sexual (arrhenotokous) reproduction normally yields haploid male and diploid female offspring. Under asexual reproduction (thelytoky), diploidized unfertilized eggs develop into females. Thelytoky is often induced by bacterial endosymbionts that achieve egg diploidization by gamete duplication. As gamete duplication leads to complete homozygosity, endosymbiont-induced thelytokous reproduction is presumed to be incompatible with CSD, which relies on heterozygosity for female development. Previously, we excluded CSD in four Asobara (Braconidae) species and proposed a two-step mechanism for Wolbachia-induced thelytoky in Asobara japonica. Here, we conclusively reject CSD in two cynipid wasp species, Leptopilina heterotoma and Leptopilina clavipes. We further show that thelytoky in L. clavipes depends on Wolbachia titer but that diploidization and feminization steps cannot be separated, unlike in A. japonica. We discuss what these results reveal about the sex determination mechanism of L. clavipes and the presumed incompatibility between CSD and endosymbiont-induced thelytoky in the Hymenoptera.


Light-induced energy dissipation in iron-starved cyanobacteria: roles of OCP and IsiA proteins.
In response to iron deficiency, cyanobacteria synthesize the iron stress-induced chlorophyll binding protein IsiA. This protein protects cyanobacterial cells against iron stress. It has been proposed that the protective role of IsiA is related to a blue light-induced nonphotochemical fluorescence quenching (NPQ) mechanism. In iron-replete cyanobacterial cell cultures, strong blue light is known to induce a mechanism that dissipates excess absorbed energy in the phycobilisome, the extramembranal antenna of cyanobacteria. In this photoprotective mechanism, the soluble Orange Carotenoid Protein (OCP) plays an essential role. Here, we demonstrate that in iron-starved cells, blue light is unable to quench fluorescence in the absence of the phycobilisomes or the OCP. By contrast, the absence of IsiA does not affect the induction of fluorescence quenching or its recovery. We conclude that in cyanobacteria grown under iron starvation conditions, the blue light-induced nonphotochemical quenching involves the phycobilisome OCP-related energy dissipation mechanism and not IsiA. IsiA, however, does seem to protect the cells from the stress generated by iron starvation, initially by increasing the size of the photosystem I antenna. Subsequently, the IsiA converts the excess energy absorbed by the phycobilisomes into heat through a mechanism different from the dynamic and reversible light-induced NPQ processes.


Implementing SBIRT (Screening, Brief Intervention and Referral to Treatment) in primary care: lessons learned from a multi-practice evaluation portfolio.
Screening, Brief Intervention and Referral to Treatment (SBIRT) is a public health framework approach used to identify and deliver services to those at risk for substance-use disorders, depression, and other mental health conditions. Primary care is the first entry to the healthcare system for many patients, and SBIRT offers potential to identify these patients early and assist in their treatment. There is a need for pragmatic "best practices" for implementing SBIRT in primary care offices geared toward frontline providers and office staff. Ten primary care practices were awarded small community grants to implement an SBIRT program in their location. Each practice chose the conditions for which they would screen, the screening tools, and how they would provide brief intervention and referral to treatment within their setting. An evaluation team communicated with each practice throughout the process, collecting quantitative and qualitative data regarding facilitators and barriers to SBIRT success. Using the editing method, the qualitative data were analyzed and key strategies for success are detailed for implementing SBIRT in primary care. The SBIRT program practices included primary care offices, federally qualified health centers, school-based health centers, and a safety-net emergency department. Conditions screened for included alcohol abuse, drug abuse, depression, anxiety, child safety, and tobacco use. Across practices, 49,964 patients were eligible for screening and 36,394 pre-screens and 21,635 full screens were completed. From the qualitative data, eight best practices for primary care SBIRT are described: Have a practice champion; Utilize an interprofessional team; Define and communicate the details of each SBIRT step; Develop relationships with referral partners; Institute ongoing SBIRT training; Align SBIRT with the primary care office flow; Consider using a pre-screening instrument, when available; and Integrate SBIRT into the electronic health record. SBIRT is an effective tool that can empower primary care providers to identify and treat patients with substance use and mental health problems before costly symptoms emerge. Using the pragmatic best practices we describe, primary care providers may improve their ability to successfully create, implement, and sustain SBIRT in their practices.


Personal resiliency: serious diagnosis and prognosis with unexpected quality outcomes.
In this grounded theory study we set out to identify what was common in stories of people with serious disease who had less than a 10% chance of survival, and who had a good quality of life at the time of first interview. A core category of personal resiliency was the organizing theme. This was a way of being and acting in the world that had the person strongly connected to life through relationships and a quality-of-life experience that made their illness secondary to their living. Whereas individual participants might not have had this sense of resiliency at the beginning of their illness, they developed it during the time they were ill, both prior to and during their recovery. Resiliency has five dimensions: Connectedness to their social environment, to family, to their physical environment, to their sense of inner wisdom (experiential spirituality), and a personal psychology with a supportive mindset and way of living which supported their values.


Sub-optimal CD4 T-lymphocyte responses among HIV infected patients who develop TB during the first year of ART.
STUDY BACKGROUND: Poor CD4 T-lymphocyte responses to anti-retroviral treatment (ART) are associated with increased HIV disease progression and mortality. In sub-Saharan Africa a substantial proportion of HIV infected patients are co-infected with TB. This study evaluated the effect of active TB presenting after ART initiation on immunological responses to ART. METHODS: A retrospective cohort study was conducted of patients initiated on ART in a South African academic hospital between 1 January 2004 and 15 May 2008. Changes in CD4 T-lymphocyte count, virological suppression and incident TB episodes occurring in the first year of ART were assessed. Sub-optimal CD4 responses were defined as 'failure to increase CD4 T-lymphocyte count by 50cells/μl at 6 month on ART'. RESULTS: The cohort for analysis included 691 patients. 141 (20.4%) had sub-optimal CD4 responses at 6 months on ART. 49 patients (7.1%) developed incident TB within the first 12 months of ART. After adjustment for age, sex, baseline CD4 count and detectable viral load, patients with incident TB were found to have a 2.20 times greater odds of a sub-optimal CD4 response at 6 month of ART as compared to those who were TB free (95%CI: 1.14-4.23). CONCLUSION: Incident TB was associated with a poor CD4 response during early ART in this cohort. Although the direction of causality cannot be determined from these data, these findings provide additional support for the initiation of ART at higher CD4 counts.


Geographic distribution and morphometric differentiation of Triatoma nitida usinger 1939 (Hemiptera: Reduviidae: Triatominae) in Guatemala.
Triatoma nitida was found in 14 (0.4%) out of 3,726 houses located in six departments across Guatemala, which were surveyed from 1994 to 1998 by the man-hour collection method. Compared to previous information, the distribution of T. nitida in Guatemala has increased from five to nine departments; the species is present in mild climates at altitudes from 960 to 1,500 m. Fourteen percent of the intradomestic T. nitida were infected with Trypanosoma cruzi. The species was often found in conjunction with other triatomines (T. dimidiata and Rhodnius prolixus). The domestic and peridomestic presence of T. nitida in Guatemala was rare, but occasionally this species was colonizing human-made constructions. T. nitida appears to have a low importance as Chagas disease vector in Guatemala, as indicated by its scarce presence in the domestic habitats and defecation patterns. However, it clearly has potential to become a Chagas vector so we recommend an on-going study of the intradomestic presence of T. nitida following the control programs in Guatemala. Morphometric analysis of 47 T. nitida males from three localities showed quantitative differences between the populations, which indicates that geographic distance is an important factor in the structuring of T. nitida populations.


Structural basis for Arl1-dependent targeting of homodimeric GRIP domains to the Golgi apparatus.
Golgins are large coiled-coil proteins that play a role in Golgi structure and vesicle traffic. The Arf-like GTPase Arl1 regulates the translocation of GRIP domain-containing golgins to Golgi membranes. We report here the 1.7 A resolution structure of human Arl1-GTP in a complex with the GRIP domain of golgin-245. The structure reveals that the GRIP domain consists of an S-shaped arrangement of three helices. The domain forms a homodimer that binds two Arl1-GTPs using two helices from each monomer. The structure is consistent with golgin-245 forming parallel coiled-coils and suggests how Arl1-GTP/GRIP complexes interact with Golgi membranes via the N termini of Arl1-GTP and the C-terminal tails of the GRIP domains. In cells, bivalent association with Arl1-GTP would increase residence time of the golgins on Golgi membranes. Despite no conservation of sequence, topology, or even helical direction, several other effectors form similar interactions with small GTPases via a pair of alpha helices, suggesting a common structural basis for effector recognition.


Glucagon-like peptide 1 induces differentiation of islet duodenal homeobox-1-positive pancreatic ductal cells into insulin-secreting cells.
Glucagon-like peptide-1 (GLP-1) is an incretin hormone capable of restoring normal glucose tolerance in aging glucose-intolerant Wistar rats. Whether the antidiabetic properties of GLP-1 are exclusively due to its insulin secretory activity remains to be determined. A GLP-1-dependent differentiation of pancreatic precursor cells into mature beta-cells has recently been proposed. The aim of this study was to investigate whether pancreatic ductal epithelial cells could be differentiated into insulin-secreting cells by exposing them to GLP-1. Rat (ARIP) and human (PANC-1) cell lines, both derived from the pancreatic ductal epithelium, were used to test this hypothesis. A major difference distinguishes these two cell lines: whereas ARIP cells spontaneously express the beta-cell differentiation factor islet duodenal homeobox-1 (IDX-1), PANC-1 cells are characteristically IDX-1 negative. GLP-1 induced the differentiation of ARIP cells into insulin-synthesizing cells, although it did not affect the phenotype of PANC-1 cells, as determined by fluorescence-activated cell sorting (FACS) analysis. Differentiation of ARIP cells by exposure to human GLP-1 occurs in a time- and dose-dependent manner, and this is associated with an increase in IDX-1 and insulin mRNA levels. Secretion of insulin was also induced in a parallel manner, and it was regulated by the concentration of glucose in the culture medium. Interestingly, PANC-1 cells, when stably transfected with human IDX-1, gained responsiveness to GLP-1 and were able to differentiate into beta-cells, as determined by FACS analysis, insulin gene expression, intracellular insulin content, and insulin accumulation in the culture medium. Finally, we demonstrated that the receptor for GLP-1 is constitutively expressed by ARIP and PANC-1 cells and that the mRNA level for this transcript was increased by cellular transfection with human IDX-1. In summary, our study provides evidence that GLP-1 is a differentiation factor for pancreatic ductal cells and that its effect requires the expression of IDX-1.


Single-Dose Tafenoquine to Prevent Relapse of Plasmodium vivax Malaria.
Treatment of Plasmodium vivax malaria requires the clearing of asexual parasites, but relapse can be prevented only if dormant hypnozoites are cleared from the liver (a treatment termed "radical cure"). Tafenoquine is a single-dose 8-aminoquinoline that has recently been registered for the radical cure of P. vivax. This multicenter, double-blind, double-dummy, parallel group, randomized, placebo-controlled trial was conducted in Ethiopia, Peru, Brazil, Cambodia, Thailand, and the Philippines. We enrolled 522 patients with microscopically confirmed P. vivax infection (>100 to <100,000 parasites per microliter) and normal glucose-6-phosphate dehydrogenase (G6PD) activity (with normal activity defined as ≥70% of the median value determined at each trial site among 36 healthy male volunteers who were otherwise not involved in the trial). All patients received a 3-day course of chloroquine (total dose of 1500 mg). In addition, patients were assigned to receive a single 300-mg dose of tafenoquine on day 1 or 2 (260 patients), placebo (133 patients), or a 15-mg dose of primaquine once daily for 14 days (129 patients). The primary outcome was the Kaplan-Meier estimated percentage of patients who were free from recurrence at 6 months, defined as P. vivax clearance without recurrent parasitemia. In the intention-to-treat population, the percentage of patients who were free from recurrence at 6 months was 62.4% in the tafenoquine group (95% confidence interval [CI], 54.9 to 69.0), 27.7% in the placebo group (95% CI, 19.6 to 36.6), and 69.6% in the primaquine group (95% CI, 60.2 to 77.1). The hazard ratio for the risk of recurrence was 0.30 (95% CI, 0.22 to 0.40) with tafenoquine as compared with placebo (P<0.001) and 0.26 (95% CI, 0.18 to 0.39) with primaquine as compared with placebo (P<0.001). Tafenoquine was associated with asymptomatic declines in hemoglobin levels, which resolved without intervention. Single-dose tafenoquine resulted in a significantly lower risk of P. vivax recurrence than placebo in patients with phenotypically normal G6PD activity. (Funded by GlaxoSmithKline and Medicines for Malaria Venture; DETECTIVE ClinicalTrials.gov number, NCT01376167 .).


Health-related quality of life following treatment for extremity soft tissue sarcoma.
The primary objective of this study was to estimate the change in health-related quality of life (HRQL) 1 year following treatment for extremity soft tissue sarcoma (STS), measured by the EQ-5D. Secondary objectives included determining clinical variables associated with HRQL at 1 year, estimating the proportion with a clinically important difference (CID) in HRQL, and evaluating variability within EQ-5D domains. Patients over the age of 16 years, treated for a localized extremity STS, were included. The EQ-5D change score from pre-treatment to 1-year follow-up was determined. The association of clinical variables with EQ-5D scores was estimated using a linear regression model. The proportion of patients with a CID in HRQL score was determined. A vector analysis of the EQ-5D domains was undertaken. The mean EQ-5D change score was 0.02. Age, sex, disease status, and initial EQ-5D score were associated with EQ-5D score at 1 year. There was a CID improvement in 32% and a deterioration in 24%. The anxiety and depression domain demonstrated the most change between baseline and 1 year after treatment. Most patients maintain a high level of HRQL following treatment for extremity STS. J. Surg. Oncol. 2016;114:821-827. © 2016 2016 Wiley Periodicals, Inc.


[Arrhythmias in adults with congenital heart disease].
Patients with surgical correction of congenital cardiopathies have a high incidence of macro-reentrant arrhythmias. In previous reports the incidence of atrial fibrillation or flutter is around 20% preoperatively and increases to 10% more after surgery. In Mustard and Senning procedures the incidence could be as high as 30%. The physiopathology of these arrhythmias is due to conduction block and heterogeneity of refractory periods due to scaring and fibrosis left by the surgical procedure. Radiofrequency ablation is a good treatment option in this patients, but with conventional approaches the percentage of success is lower and with higher recurrence. In our institution out of 39 patients with macro-reentrant atrial tachycardia, acute success was 77% in patients with isthmus dependent flutter and 44% if the Isthmus was not part of the circuit. Recurrence in both groups was 42%. New mappings systems like Localisa, CARTO an NavX, are useful to localize areas of scar and block, that produce multiple conduction channels that can participate in reentrant arrhythmias. Radiofrequency ablation of these channels is up to day the ideal approach for these patients. Arrhythmias in patients with congenital cardiopathies are frequent and complicate the evolutions of these patients. Radiofrequency ablation is the treatment of choice in centers with experience. The use of non fluoroscopic electroanatomic mapping systems is of great help in this setting.


[The current methods for predicting the course of rheumatoid arthritis].
One of the lines of inquiry into rheumatoid arthritis (RA) is a quest for markers capable of prognosticating the course of the illness. It is only genetic factors, the presence of rheumatoid factor, and activity of the illness that are most significant prognostic criteria. Showing much promise among the current approaches is DAS (Disease Activity Score) which relies on four variables (Richey index, number of swollen joints, ESR, and health index). The baseline and cumulative DAS values over a certain period of time can be used as prognostic factors. It is these criteria that the European League Against the Rheumatic Diseases Committee on Standardization of Clinical and Therapeutic Investigations have given approval to. They have been termed EULAR criteria.


EPSPS gene amplification in glyphosate-resistant Bromus diandrus.
Glyphosate is the most widely used herbicide in the world and has been intensively used to control B. diandrus, a problematic weed of crops and pastures in southern Australia. Resistance to glyphosate was identified in two populations of B. diandrus that were nearly fivefold more resistant to glyphosate than wild-type plants. Both populations contained EPSPS gene amplification, with resistant plants having an average of around 20-fold the number of copies of EPSPS compared with susceptible plants. EPSPS expression was also increased in resistant plants of both populations; however, expression levels were not correlated with the number of EPSPS copies. Amplification of only one of the four EPSPS genes present in B. diandus was detected. Investigation into the inheritance of glyphosate resistance found no segregation in the F2 generation. Every individual in the F2 populations contained between three and 30 copies of EPSPS; however, on average they contained fewer copies compared with the parent resistant population. Glyphosate resistance in B. diandrus is due to EPSPS gene amplification. Resistance is heritable but complex.


Cancer Screening Differences Among Muslims and Non-Muslims: Insights from the Chicago Multiethnic Prevention and Surveillance Study.
While cancer screening disparities along socioeconomic and racial/ethnic lines are well studied, differences based on religious affiliation are under-researched. Though diverse in terms of race/ethnicity, Muslim Americans appear to share values and beliefs that similarly inform their health and healthcare seeking behaviors. Cancer screening disparities among Muslim Americans are also understudied. To examine differences in cancer screening behaviors based on Muslim affiliation, we analyzed data from a longitudinal cohort study examining lifestyle, healthcare access, environmental, and genetic factors on the health of Chicagoans. Of 7552 participants, 132 (1.7%) were Muslim. Between Muslim and non-Muslims, there were no significant differences in prostate, cervical, and breast cancer screening rates, but Muslims were less likely to undergo colorectal cancer screening. When differences in obesity and insurance status were accounted for in a multivariate regression model, religious affiliation was no longer significantly associated with screening rates. Religious values can influence cancer screening behaviors; hence, tracking cancer screening along religious lines may illuminate previously unknown disparities. Our analysis of a predominately African American cohort of Chicagoans, however, did not reveal religious affiliation to predict cancer screening disparities.


Effects of nitric oxide stimulation on the brain.
Nitric oxide (NO) is known as the neurotransmitter responsible for penile erection. Recent experimental studies showed that NO is recognized as an important messenger mediating male copulatory behavior and penile erection in the central nervous system. In this article, the roles of central NO on modulation of male copulatory behavior and penile erection in male rats are reviewed mainly based on our previous experimental results. We focused on two brain areas in the hypothalamus for this purpose: the medial preoptic area (MPOA) and paraventricular nucleus (PVN). Increase in extracellular NO level in the MPOA facilitates copulatory behaviors, while decreases in the NO level reduce them. The altered NO level in the PVN increases frequency of reflexive erections, but does not affect copulatory behavior. Taken together, the current results and previous reports suggest that central NO may modulate male sexual functions and NO in two distinct brain areas and may play different roles through activation of different neurotransmitters. Activities of NO in the hypothalamus are affected by the existence of gonadal steroids. Moreover, our experimental results clearly showed that the extracellular NO level in the MPOA and PVN decreased with aging, and testosterone replacement in aged rats restored NO levels in both brain areas. These results postulate that changes of central NO with aging and following hormone replacement might partly contribute to changes in male rats sexual behavior corresponding to aging and replacement. Moreover, our recent report demonstrated that long-term testosterone replacement in aged rats is more effective for restoration of sexual behavior, which may be partly induced by central NO, than high-dose, short-term replacement.


Causes of death abroad: analysis of data on bodies returned for cremation to Scotland.
The majority of travelers from the UK and Scotland visit Europe, particularly Spain and France, as well as North America, yet surveillance that aids in travel medicine guidance continues to focus on infectious diseases relating to developing countries. Here, we report on causes of death among all bodies returned to Scotland for cremation. Data collected by the Scottish Government on bodies being returned from abroad for cremation was collated for the period 2000 to 2004, and analyzed to identify the cause and location of death among travelers as well as to test the hypothesis that for death due to failure of the circulatory system among Scots there was a significant association between age at death and whether death occurred in Scotland or abroad. Of the 572 deaths reported between 2000 and 2004, 73% occurred in the European region and 10% in the Americas. With respect to the cause, trauma accounted for 20.4%, infectious diseases 1.5%, and other non-infectious causes accounted for 75.5% of deaths. Among the latter, the major cause of death was due to failure of the circulatory system (77.0%). A significant association was observed between death abroad due to failure of the circulatory system and younger age at death for all (χ(2) = 26.9, df = 3, p < 0.001) and for males (χ(2) = 20.7, df = 3, p < 0.001) but not for females (χ(2) = 2.7, df = 1, p = 0.099). The data indicates a low rate of death among Scots traveling abroad, with trauma and other non-infectious causes being the most common cause of death; failure of the circulatory system was the most common cause of death in the latter group. Europe and the Americas were the most common locations of death. Although travel health services should continue to advise travelers to developing countries on infectious disease risks, it is also important that travel health acts as venue for providing key advice and preventative means to all travelers, including those to developed countries. Those agencies, organizations, and companies who deal with travelers along their journey should also engage with travel health experts and practitioners to reduce the risk of adverse outcomes, including death, to travelers.


Effects of lipopolysaccharide on leptin transport across the blood-brain barrier.
Leptin is a 16-kDa protein secreted by fat cells and transported into the brain where it decreases appetite and increases body temperature. Leptin transport is saturable and regulated by epinephrine, triglycerides, and starvation. Lipopolysaccharide (LPS) is derived from bacterial cell walls and also decreases appetite and increases body temperature. LPS is known to increase leptin levels in serum and to affect the passage of other regulatory proteins across the blood-brain barrier (BBB). Here, we examined the ability of LPS, at doses which induce weight loss, to modify the BBB transport of radioactive leptin (I-Lep). The transport rate of intravenously injected I-Lep was decreased by 50-60% from 8 to 12 h after a single i.p. injection of LPS (3 mg/kg). The effect of LPS was dose-dependent. In comparison to the brain/serum ratio, the baseline cerebrospinal fluid (CSF)/serum ratio for I-Lep was much lower and not inhibited by LPS. LPS did not affect I-Lep transport when studied by the brain perfusion method nor was Ob-Ra mRNA expression in isolated brain microvessels altered, demonstrating that a circulating factor rather than altered BBB function was responsible for inhibition. Brain perfusion showed that LPS was not this factor. Serum leptin was doubled and serum triglycerides increased by 44% after LPS administration, suggesting these to be the circulating inhibitory factors. In conclusion, a single dose of LPS has long-lasting effects on the transport of serum leptin across the BBB that are likely mediated through self-inhibition and triglycerides.