[Transperitoneal laparoscopic adrenalectomy: results after 70 procedures].
To present our surgical experience in laparoscopic adrenalectomy after 70 procedures, comparing our results with the published series and analyzing the learning curve. Between June 1997 and December 2007 we performed 87 adrenalectomies (70 transperitoneal laparoscopy, 1 retroperitoneoscopy and 16 open surgery); 60% were female. Mean age was 54 years 15-80); 35 were left adrenalectomies, 33 right and one bilateral. Mean mass size was 3,7 cm (1-10). All patients were assessed regarding the following variables: preoperative diagnosis, histopathology, operative time, blood loss, complications, conversion to open surgery and hospital stay. We compared the results of the first 40 surgical procedures (Group 1) and the last 30 (Group 2). We also compared our re-sults with published series. Mean time follow-up was 12 months (2-60). The most frequent preoperative diagnoses were: active suprarenal mass (50%), incidentaloma (20%), pheochromocytoma (20%). Histopathology confirmed 60% adenomas, 14% pheochromocytomas and 11% cortical nodular hyperplasia. Mean operative time was 161 minutes (60-300). Average blood loss (hematocrit) was 5.2 (1-10). Conversion and complication rate were 8.5 and 7%, respectively. Mean hospital stay was 4 days (2-30). In group 2, only reduction on mean operative time (120 min) and hospital stay (3 days) was statistically significant. Transperitoneal laparoscopic adrenalectomy is a feasible and safe procedure in urological groups with previous laparoscopic experience. Our results are similar to the published series and confirm the efficacy, safety and reproducibility of this technique.


Functional analyses of the interaction of chicken interleukin 23 subunit p19 with IL-12 subunit p40 to form the IL-23 complex.
This study represents the first description of the cloning of chicken IL-23p19 (ChIL-23α) and the function of the IL-23 complex in birds. Multiple alignment of ChIL-23α with other known IL-23α amino acid sequences revealed regions of amino acid conservation. The homologies of ChIL-23α, IL-12p35, and similar mammalian subunits ranged between 26% and 42%. ChIL-23α consisted of four exons and three introns; similar to those in humans and mice, and limited conservation of synteny between the human and chicken genomes was observed. Using bioinformatics tools, we identified the NF-κB, C/EBPα-β, c-Jun, c-Rel, AP-1, GATA-1, and ER promoter sites in ChIL-23α. Moreover, IL-23α mRNA was more highly expressed than IL-12p40 and IL-12p35 mRNA in several organs of chickens infected with Salmonella. In addition, ChIL-23 complex are associated with IL-23R, IL-12Rβ1 receptors; activate the JAK2/TYK2, STAT1/3, SOCS1 genes, and induced proinflammatory cytokines in immune cells. Collectively, these results indicate that ChIL-23 is a member of the IL-12 family, has proinflammatory properties related to IL-23R and IL-12Rβ1 receptor expression, and activates the JAK/STAT signaling pathway that results in the interaction of ChIL-23α with ChIL-12p40 to form the novel ChIL-23 complex. Our results provide novel insights into the regulation of immunity, inflammation, and immunopathology.


Orlistat in clozapine- or olanzapine-treated patients with overweight or obesity: a 16-week open-label extension phase and both phases of a randomized controlled trial.
To explore long-term effects of orlistat in adult clozapine- or olanzapine-treated patients with DSM-IV-diagnosed schizophrenia and overweight or obesity who tolerate orlistat. Orlistat or placebo was added to clozapine or olanzapine in stable doses in a 16-week randomized controlled trial. Open-label orlistat was added to the antipsychotics during a 16-week extension phase for those completing the double-blind phase. No low-calorie diet or participation in behavioral programs was required. Body weight (primary outcome) and some metabolic parameters were measured prospectively. Analyses were performed for those completing both phases (ie, population differing from that reported earlier). The study was conducted from 2004 through 2005. During the open-label phase, the 44 patients experienced mean ± SD body weight loss of -1.29 ± 3.04 kg, P = .007. During both phases, men (but not women) showed a weight loss of -2.39 ± 5.45 kg, P = .023. Some subgroups showed desirable changes in several metabolic parameters. Prolonged (32 weeks) orlistat treatment yielded no additional benefits as compared to short (16 weeks) treatment. In clozapine- or olanzapine-treated overweight or obese patients able to take orlistat on a long-term basis, the drug, with no concomitant hypocaloric diet or behavioral interventions, caused moderate weight loss only in men. However, some metabolic benefits may be achieved independently of weight changes. In patients who do not respond to orlistat within the first 16 weeks, continuation treatment may provide no additional benefits. controlled-trials.com Identifier: ISRCTN65731856.


Older age does not impact perioperative complications after robot-assisted radical prostatectomy.
To determine whether men aged 70 years and older had more perioperative complications after robot-assisted radical prostatectomy (RARP) compared with younger patients, a retrospective review was performed on patients who underwent RARP between March 2004 and September 2009. Subjects were stratified according to age into four groups (age 30-49, 50-59, 60-69, and ≥70 years). American Society of Anesthesiologists (ASA) scores were obtained. Complication rates in the perioperative period, transfusion rates, and length of stay were compared. Complications were classified using the previously validated Clavien system. There were a total of 293 patients aged 70 years and older amongst the 1,223 total subjects. ASA comorbidity scores did vary significantly amongst the different age groups, and there was modest correlation noted between ASA and age. There was no statistically significant difference amongst complication rates in men aged 70 years and older (15%) compared with the other cohorts (P = 0.832). There was also no significant difference in transfusion rates (P = 0.170) or length of stay (P = 0.131). Patients with higher ASA scores (ASA 3-4) had more Clavien I-II complications compared with patients with ASA scores of 1-2 (15.5% versus 10.3%, P = 0.03). There was no difference in transfusion rates or length of stay between the ASA scores. There are no more complications in men aged 70 years and older compared with men <70 years of age undergoing robot-assisted radical prostatectomy. RARP is a safe treatment option to offer to the selected elderly patient.


Deletion of FRT-sites by no-SCAR recombineering in Escherichia coli.
Lambda-Red recombineering is the most commonly used method to create point mutations, insertions or deletions in Escherichia coli and other bacteria, but usually an Flp recognition target (FRT) scar-site is retained in the genome. Alternative scarless recombineering methods, including CRISPR/Cas9-assisted methods, generally require cloning steps and/or complex PCR schemes for specific targeting of the genome. Here we describe the deletion of FRT scar-sites by the scarless Cas9-assisted recombineering method no-SCAR using an FRT-specific guide RNA, sgRNAFRT, and locus-specific ssDNA oligonucleotides. We applied this method to construct a scarless E. coli strain suitable for gradual induction by l-arabinose. Genome sequencing of the resulting strain and its parent strains demonstrated that no additional mutations were introduced along with the simultaneous deletion of two FRT scar-sites. The FRT-specific no-SCAR selection by sgRNAFRT/Cas9 may be generally applicable to cure FRT scar-sites of E. coli strains constructed by classical λ-Red recombineering.


Who can best recruit to randomized trials? Randomized trial comparing surgeons and nurses recruiting patients to a trial of treatments for localized prostate cancer (the ProtecT study).
Recruitment to randomized trials is often difficult, but few studies have investigated interventions to improve recruitment. In a randomized trial nested within a trial of treatments for localized prostate cancer, we investigated the comparative effectiveness and cost-effectiveness of nurses and surgeons in recruiting patients. Men with localized prostate cancer were randomized to see a nurse or urologic surgeon for an "information appointment" in which they were asked to consent to the ProtecT treatment trial comparing surgery, radiotherapy, and active monitoring. Analysis was conducted by intention to treat using chi-square with 95% confidence intervals for proportions and differences between groups. An economic evaluation was performed using the duration of appointments and grade of recruitment staff. Case-finding identified 167 men with localized prostate cancer. One hundred fifty (90%) took part in the recruitment trial. There was a 4.0% difference between nurses and surgeons in recruitment rates (67% nurses, 71% urologists, 95% CI -10.8% to +18.8%, P=.60). Cost-minimization analysis showed that nurses spent longer times with patients but surgeon costs were higher and nurses often supported surgeon-led clinics. Nurses were as effective and more cost-effective recruiters than urologic surgeons. This suggests an increased role for nurses in recruiting patients to randomized trials.


Histone lysine methyltransferases and demethylases in Plasmodium falciparum.
Dynamic histone lysine methylation, regulated by methyltransferases and demethylases, plays fundamental roles in chromatin structure and gene expression in a wide range of eukaryotic organisms. A large number of SET-domain-containing proteins make up the histone lysine methyltransferase (HKMT) family, which catalyses the methylation of different lysine residues with relatively high substrate specificities. Another large family of Jumonji C (JmjC)-domain-containing histone lysine demethylases (JHDMs) reverses histone lysine methylation with both lysine site and methyl-state specificities. Through bioinformatic analysis, at least nine SET-domain-containing genes were found in the malaria parasite Plasmodium falciparum and its sibling species. Phylogenetic analysis separated these putative HKMTs into five subfamilies with different putative substrate specificities. Consistent with the phylogenetic subdivision, methyl marks were found on K4, K9 and K36 of histone H3 and K20 of histone H4 by site-specific methyl-lysine antibodies. In addition, most SET-domain genes and histone methyl-lysine marks displayed dynamic changes during the parasite asexual erythrocytic cycle, suggesting that they constitute an important epigenetic mechanism of gene regulation in malaria parasites. Furthermore, the malaria parasite and other apicomplexan genomes also encode JmjC-domain-containing proteins that may serve as histone lysine demethylases. Whereas prokaryotic expression of putative active domains of four P. falciparum SET proteins did not yield detectable HKMT activity towards recombinant P. falciparum histones, two protein domains expressed in vitro in a eukaryotic system showed HKMT activities towards H3 and H4, respectively. With the discovery of these Plasmodium SET- and JmjC-domain genes in the malaria parasite genomes, future efforts will be directed towards elucidation of their substrate specificities and functions in various cellular processes of the parasites.


ACKR3-arrestin2/3 complexes reveal molecular consequences of GRK-dependent barcoding.
Atypical chemokine receptor 3 (ACKR3, also known as CXCR7) is a scavenger receptor that regulates extracellular levels of the chemokine CXCL12 to maintain responsiveness of its partner, the G protein-coupled receptor (GPCR), CXCR4. ACKR3 is notable because it does not couple to G proteins and instead is completely biased towards arrestins. Our previous studies revealed that GRK2 and GRK5 install distinct distributions of phosphates (or "barcodes") on the ACKR3 carboxy terminal tail, but how these unique barcodes drive different cellular outcomes is not understood. It is also not known if arrestin2 (Arr2) and 3 (Arr3) bind to these barcodes in distinct ways. Here we report cryo-electron microscopy structures of Arr2 and Arr3 in complex with ACKR3 phosphorylated by either GRK2 or GRK5. Unexpectedly, the finger loops of Arr2 and 3 directly insert into the detergent/membrane instead of the transmembrane core of ACKR3, in contrast to previously reported "core" GPCR-arrestin complexes. The distance between the phosphorylation barcode and the receptor transmembrane core regulates the interaction mode of arrestin, alternating between a tighter complex for GRK5 sites and heterogenous primarily "tail only" complexes for GRK2 sites. Arr2 and 3 bind at different angles relative to the core of ACKR3, likely due to differences in membrane/micelle anchoring at their C-edge loops. Our structural investigations were facilitated by Fab7, a novel Fab that binds both Arr2 and 3 in their activated states irrespective of receptor or phosphorylation status, rendering it a potentially useful tool to aid structure determination of any native GPCR-arrestin complex. The structures provide unprecedented insight into how different phosphorylation barcodes and arrestin isoforms can globally affect the configuration of receptor-arrestin complexes. These differences may promote unique downstream intracellular interactions and cellular responses. Our structures also suggest that the 100% bias of ACKR3 for arrestins is driven by the ability of arrestins, but not G proteins, to bind GRK-phosphorylated ACKR3 even when excluded from the receptor cytoplasmic binding pocket.


Alignment of 1000 Genomes Project reads to reference assembly GRCh38.
The 1000 Genomes Project produced more than 100 trillion basepairs of short read sequence from more than 2600 samples in 26 populations over a period of five years. In its final phase, the project released over 85 million genotyped and phased variants on human reference genome assembly GRCh37. An updated reference assembly, GRCh38, was released in late 2013, but there was insufficient time for the final phase of the project analysis to change to the new assembly. Although it is possible to lift the coordinates of the 1000 Genomes Project variants to the new assembly, this is a potentially error-prone process as coordinate remapping is most appropriate only for non-repetitive regions of the genome and those that did not see significant change between the two assemblies. It will also miss variants in any region that was newly added to GRCh38. Thus, to produce the highest quality variants and genotypes on GRCh38, the best strategy is to realign the reads and recall the variants based on the new alignment. As the first step of variant calling for the 1000 Genomes Project data, we have finished remapping all of the 1000 Genomes sequence reads to GRCh38 with alternative scaffold-aware BWA-MEM. The resulting alignments are available as CRAM, a reference-based sequence compression format. The data have been released on our FTP site and are also available from European Nucleotide Archive to facilitate researchers discovering variants on the primary sequences and alternative contigs of GRCh38.


Embedding implant dentistry into the undergraduate dental curriculum.
This article represents the consensus from a meeting of the British Society of Prosthetic Dentistry's Education Group which met to discuss implant dentistry at the undergraduate student level. The consensus recognises the need to embed the teaching of implant supported prostheses if such a treatment modality is to be widely practised after graduation. This short article makes some recommendations as well as detailing some challenges and barriers that need to be overcome if implant dentistry is to be successfully embedded in an undergraduate dental curriculum.


High-throughput sequencing defines donor and recipient HLA B-cell epitope frequencies for prospective matching in transplantation.
Compatibility for human leukocyte antigen (HLA) genes between transplant donors and recipients improves graft survival but prospective matching is rarely performed due to the vast heterogeneity of this gene complex. To reduce complexity, we have combined next-generation sequencing and in silico mapping to determine transplant population frequencies and matching probabilities of 150 antibody-binding eplets across all 11 classical HLA genes in 2000 ethnically heterogeneous renal patients and donors. We show that eplets are more common and uniformly distributed between donors and recipients than the respective HLA isoforms. Simulations of targeted eplet matching shows that a high degree of overall compatibility, and perfect identity at the clinically important HLA class II loci, can be obtained within a patient waiting list of approximately 250 subjects. Internal epitope-based allocation is thus feasible for most major renal transplant programs, while regional or national sharing may be required for other solid organs.


[Clinical and morphological features of infectious endocarditis in HIV-infected intravenous drug abusers].
The aim of this work was to study the clinical course and morphogenesis of infectious endocarditis in HIV-infected intravenous drug abusers. In this patients, endocarditis was an acute disease with high frequency of thromboembolic complications and ill-defined exudative component of the inflammatory reaction. Autopsies dated to 1993-2008 showed that neither the presence of morphological signs of early HIV infection nor chronic hepatitis C with minimal or moderate inflammation has an appreciable effect on the intrahospital mortality rate in these patients.


Plk4-dependent phosphorylation of STIL is required for centriole duplication.
Duplication of centrioles, namely the formation of a procentriole next to the parental centriole, is regulated by the polo-like kinase Plk4. Only a few other proteins, including STIL (SCL/TAL1 interrupting locus, SIL) and Sas-6, are required for the early step of centriole biogenesis. Following Plk4 activation, STIL and Sas-6 accumulate at the cartwheel structure at the initial stage of the centriole assembly process. Here, we show that STIL interacts with Plk4 in vivo. A STIL fragment harboring both the coiled-coil domain and the STAN motif shows the strongest binding affinity to Plk4. Furthermore, we find that STIL is phosphorylated by Plk4. We identified Plk4-specific phosphorylation sites within the C-terminal domain of STIL and show that phosphorylation of STIL by Plk4 is required to trigger centriole duplication.


Mouse mesenchymal stem cells inhibit high endothelial cell activation and lymphocyte homing to lymph nodes by releasing TIMP-1.
Mesenchymal stem cells (MSC) represent a promising therapeutic approach in many diseases in view of their potent immunomodulatory properties, which are only partially understood. Here, we show that the endothelium is a specific and key target of MSC during immunity and inflammation. In mice, MSC inhibit activation and proliferation of endothelial cells in remote inflamed lymph nodes (LNs), affect elongation and arborization of high endothelial venules (HEVs) and inhibit T-cell homing. The proteomic analysis of the MSC secretome identified the tissue inhibitor of metalloproteinase-1 (TIMP-1) as a potential effector molecule responsible for the anti-angiogenic properties of MSC. Both in vitro and in vivo, TIMP-1 activity is responsible for the anti-angiogenic effects of MSC, and increasing TIMP-1 concentrations delivered by an Adeno Associated Virus (AAV) vector recapitulates the effects of MSC transplantation on draining LNs. Thus, this study discovers a new and highly efficient general mechanism through which MSC tune down immunity and inflammation, identifies TIMP-1 as a novel biomarker of MSC-based therapy and opens the gate to new therapeutic approaches of inflammatory diseases.


The effects of IDRA 21, a positive modulator of the AMPA receptor, on delayed matching performance by young and aged rhesus monkeys.
IDRA 21, a positive allosteric modulator of the glutamate AMPA receptor, produced a concentration-dependent inhibition of glutamate-induced inactivation of membrane currents in recombinant HEK 293 (human embryonic kidney) cells stably transfected with human GluR1/2 flip receptors. IDRA 21 doubled the charge transfer at a concentration of 70 microM, suggesting that this compound can facilitate excitatory neurotransmission via GluR 1/2 receptors. We next sought to exploit this mechanism of action by examining the drug as a potential cognition-enhancing agent in non-human primates. Oral administration of IDRA 21 produced a highly significant improvement in the performance of a delayed matching-to-sample (DMTS) task by young adult rhesus monkeys. The pattern of task improvement over the dose range 0.15-10 mg/kg was maintained to 48 hr after the single dose administration. For sessions run after administration of the individualized Best Dose of IDRA 21, task accuracy for Long delay (most difficult) trials was increased by 34% of vehicle. Animals were randomly assigned fixed doses of IDRA 21 to determine whether the positive mnemonic response could be maintained. The repeated doses were separated by 3 days, thus allowing for potential cumulative effects. IDRA 21 produced a gradual increase in task accuracy that was maintained on average above vehicle performance levels over an intermittent dosing schedule during a total period of 3 weeks. A separate group of aged monkeys (>20 y) were, as a group, impaired (during vehicle testing) in DMTS performance efficiency relative to the young cohort. IDRA 21 also improved task accuracy by aged rhesus monkeys over the same dose range, but the responses were not as robust as those exhibited by young animals. Aged subjects also appeared to be more individually sensitive to drug dose, and they exhibited shorter task latencies than did the young group. Despite these differences, when the individualized Best Doses were considered, IDRA 21 produced a robust increase in DMTS accuracy of up to 18% of vehicle for trials associated with Medium delay intervals. For both study groups, no obvious untoward effects of IDRA 21 were noted. These findings support the use of AMPA modulators like IDRA 21 in the treatment of cognitive/memory disorders, including those associated with aging. They also indicate that the drug is associated with long-term effects that could limit dosing regimens to one dose every two or three days. The nature of the protracted mnemonic effects produced by the compound remains to be elucidated.


Paradoxical effect of capecitabine in 5-fluorouracil-induced cardiotoxicity: A case vignette and literature review.
5-fluorouracil is a chemotherapeutic agent that plays an important role in the treatment of various cancers including head and neck and gastrointestinal malignancies. Therapy with 5-fluorouracil is rarely associated with cardiotoxic effects including angina, heart failure, myocardial infarction and cardiac arrest, resulting in discontinuation at the expense of sub-optimal treatment of the targeted malignancy. In this article, we review the literature reported on 5-fluorouracil-associated cardiotoxicity and present a case of a patient who experienced chest pain on 5-fluorouracil. The cardiac symptoms subsided after initiation of capecitabine, the oral formulation of 5-fluorouracil. To our knowledge, this is only the second reported case where 5-fluorouracil was successfully replaced by capecitabine without recurrence of cardiac symptoms. Capecitabine may be a viable option for patients who develop 5-fluorouracil-induced chest pain. However, large clinical trials are warranted to confirm these findings. Currently, there is insufficient evidence to recommend an optimal approach for safe and effective alternative treatment for patients who experience 5-fluorouracil-induced cardiac adverse events.


Calorie restriction attenuates LPS-induced sickness behavior and shifts hypothalamic signaling pathways to an anti-inflammatory bias.
Calorie restriction (CR) has been demonstrated to alter cytokine levels; however, its potential to modify sickness behavior (fever, anorexia, cachexia) has not. The effect of CR on sickness behavior was examined in male C57BL/6J mice fed ad libitum or restricted 25% (CR25%) or restricted 50% (CR50%) in food intake for 28 days and injected with 50 μg/kg of LPS on day 29. Changes in body temperature, locomotor activity, body weight, and food intake were determined. A separate cohort of mice were fed ad libitum or CR50% for 28 days, and hypothalamic mRNA expression of inhibitory factor κB-α (IκB-α), cyclooxygenase-2 (COX-2), prostaglandin E(2) (PGE(2)), suppressor of cytokine signaling 3 (SOCS3), IL-10, neuropeptide Y (NPY), leptin, proopiomelanocortin (POMC), and corticotrophin-releasing hormone (CRH) were determined at 0, 2, and 4 h post-LPS. CR50% mice did not develop fevers, whereas the CR25% mice displayed a fever shorter in duration but with the same peak as the controls. Both CR25% and CR50% mice showed no sign of anorexia and reduced cachexia after LPS administration. Hypothalamic mRNA expression of NPY and CRH were both increased by severalfold in CR50% animals preinjection compared with controls. The CR50% mice did not demonstrate the expected rise in hypothalamic mRNA expression of COX-2, microsomal prostaglandin E synthase-1, POMC, or CRH 2 h post-LPS, and leptin expression was decreased at this time point. Increases in SOCS3, IL-10, and IκB-α expression in CR50% animals were enhanced compared with ad libitum-fed controls at 4 h post-LPS. CR results in a suppression of sickness behavior in a dose-dependent manner, which may be due to CR attenuating proinflammatory pathways and enhancing anti-inflammatory pathways.


Cognitive-behavioral stress management intervention decreases the prevalence of depression and enhances benefit finding among women under treatment for early-stage breast cancer.
The authors tested effects of a 10-week group cognitive-behavioral stress management intervention among 100 women newly treated for Stage 0-II breast cancer. The intervention reduced prevalence of moderate depression (which remained relatively stable in the control condition) but did not affect other measures of emotional distress. The intervention also increased participants' reports that having breast cancer had made positive contributions to their lives, and it increased generalized optimism. Both remained significantly elevated at a 3-month follow-up of the intervention. Further analysis revealed that the intervention had its greatest impact on these 2 variables among women who were lowest in optimism at baseline. Discussion centers on the importance of examining positive responses to traumatic events--growth, appreciation of life, shift in priorities, and positive affect-as well as negative responses.


Age at onset of necrotizing enterocolitis. Risk factors in small infants.
We compared the features of 45 infants with early-onset necrotizing enterocolitis (NEC) (in the first ten days of life) and 41 infants with late-onset NEC (after the tenth day of life); all weighed 1,500 g or less at birth. Infants with late-onset NEC were smaller at birth and had younger gestational ages than infants with early-onset illness. No significant differences between the two groups, however, were found in sex or race distribution, mean one- and five-minute Apgar scores, or in the relative frequency of occurrence of 30 maternal and neonatal risk factors. Intravenous alimentation was administered significantly more frequently to infants with late-onset NEC. Birth weight-specific weekly attack rates for NEC declined dramatically when infants reached the equivalent of 35 to 36 weeks' gestational age. We hypothesize that the timing of noxious events in relation to gut maturity may explain these observations.


Dilated cardiomyopathy in young adult Africans: a sequel to infections?
Over a period of 6 years, only twelve cases of dilated cardiomyopathy were clinically diagnosed in Nigerians between the ages of 11-30 years at the University College Hospital, Ibadan, Nigeria. Eleven presented with heart failure, while the twelfth patient presented with a cerebrovascular accident. Two other patients also had a cerebrovascular accident. A history of febrile illness was obtained in seven, but in only three was fever unresponsive to antimalarials, documented on admission. Antistreptolysin-O titre was normal and erythrocyte sedimentation rates elevated in each of the patients. Leucocytosis was present in six, three had a four-fold rise or fall in antibody titres against Coxsackie-B viruses and one, a four-fold rise or fall against Toxoplasma gondii. Histological evidences of myopericarditis were found in three of the six patients who died. It is concluded that dilated cardiomyopathy is rare in young adult Nigerians, and that constitutional upset is common, as in children, but prognosis is poorer. Infections by Coxsackie-B viruses, T. gondii and possibly other viruses appear to be of major aetiological factors


How will the greening of the Arctic affect an important prey species and disturbance agent? Vegetation effects on arctic ground squirrels.
Increases in terrestrial primary productivity across the Arctic and northern alpine ecosystems are leading to altered vegetation composition and stature. Changes in vegetation stature may affect predator-prey interactions via changes in the prey's ability to detect predators, changes in predation pressure, predator identity and predator foraging strategy. Changes in productivity and vegetation composition may also affect herbivores via effects on forage availability and quality. We investigated if height-dependent effects of forage and non-forage vegetation determine burrowing extent and activity of arctic ground squirrels (Urocitellus parryii). We collected data on burrow networks and activity of arctic ground squirrels across long-term vegetation monitoring sites in Denali National Park and Preserve, Alaska. The implications of height-specific cover of potential forage and non-forage vegetation on burrowing behaviour and habitat suitability for arctic ground squirrels were investigated using hierarchical Bayesian modelling. Increased cover of forbs was associated with more burrows and burrow systems, and higher activity of systems, for all forb heights. No other potential forage functional group was related to burrow distribution and activity. In contrast, height-dependent negative effects of non-forage vegetation were observed, with cover over 50-cm height negatively affecting the number of burrows, systems and system activity. Our results demonstrate that increases in vegetation productivity have dual, potentially counteracting effects on arctic ground squirrels via changes in forage and vegetation stature. Importantly, increases in tall-growing woody vegetation (shrubs and trees) have clear negative effects, whereas increases in forb should benefit arctic ground squirrels.


A multimedia-based histology laboratory course: elimination of the traditional microscope laboratory.
We have developed a multimedia-based laboratory course which has enabled us to eliminate the microscope and traditional microscope laboratory that have been mainstays of our histology course and histology courses at almost all institutions where histology is taught. The multimedia laboratory uses a library of histology images (approximately 24,000) stored on videodisc ( A Photographic Atlas, by S. Downing) as its microscope slide collection and accesses those images through barcode and computer interfaces. The laboratory workstations consist of a videodisc player, videodisc monitor, computer, and computer monitor. One workstation is available for every 4-students, and our students are encouraged to work together in groups of four or five. In our current set-up, the students are introduced to and instructed in the basic principles of histology using a computer program that interfaces with the videodisc images. The computer program is divided into 19 chapters (the chapters are typical of the chapters found in a normal histology textbook) and has: (1) a laboratory component that covers the material traditionally covered in the microscope laboratory, and (2) a lecture component that enables the students to evaluate their understanding of the lecture material in a non-punishing way. The laboratory section of each chapter is divided into a "MicroLab" section, an "InFo Time" section, and a "Quiz Time" section. Each of these sections interfaces with histological images stored on the videodisc. The students are encouraged to work through the "MicroLab" section of each chapter before moving on to the "InFo Time" and "Quiz Time" sections. The "MicroLab" sections introduce the students to the various tissues and organs of the body and is interfaced with the videodisc player and the histology images stored on the videodisc. These sections describe the basic histological features of the various tissues and organs and give the students access to multiple examples of what they are studying. The "InFo Time" sections bring up specific images and ask the students to think about the images. Information about the images being observed is available if the students want it and the students can flag those images that they found difficult. The Quiz Time section of the program is also interfaced with the videodisc player and provides access to a large number of histology images stored on the videodisc. The "Quiz Time" sections provide non-punishing review questions that the students can study after she has worked her way through the "MicroLab" and "InFo Time" sections. In addition to the use of a computer program to access the histology images stored on videodisc, we use barcodes that address specific images on the histology videodisc in a variety of ways to augment the students' laboratory and lecture experience. The benefits of using multimedia in place of the traditional microscope and microscope slide collection are numerous and include the speed at which specific histological images can be accessed and reviewed (when compared to finding a structure on a glass slide), a significant reduction in the amount of laboratory time needed by the student to learn the same amount of information, the ease of tutoring on a large monitor screen (when compared to trying to discuss a histological structure with a student through the eyepiece of a microscope), the encouragement of group study (which is difficult to do when a student is working 1-on-with a microscope), and the reduction of the number of faculty necessary to cover a typical histology laboratory session. The use of barcodes that address specific videodisc histology images has greatly changed our examination procedures and has significantly expanded the usefulness of the traditional lecture note handouts given to our students.


Charge microheterogeneity of the major capsid protein of polyoma virus.
The behavior in isoelectric focusing of the major capsid polypeptide VPI of several strains of polyoma virus was studied. Two previously recognized phenomena were reexamined, namely, (i) the separation of the VP1 polypeptide into multiple subspecies differing only slightly from each other in apparent isoelectric point and (ii) strain differences in the overall apparent net charge of the family of VP1 subspecies. It was found that the pattern of subspecies was reproducible when focusing was initiated from either the basic or acidic region of the gel, keeping the ampholyte mixture constant. However, individual subspecies were unstable, and labeled polypeptide could be shifted dramatically by either refocusing of separated subspecies or by altering the concentration of ampholytes. These findings suggest that protein-protein and protein-ampholyte interactions play an important role in the generation of this charge heterogeneity. The basis for the overall charge difference between the VP1 of 3049 virus and several other strains (lpD, lpS, ts59, and A2) was studied, using recombinant viruses constructed of specific sequences derived from 3049 and lpD genomes. The portion of the VP1 polypeptide carrying the altered charge could be mapped to the body of the molecule 3' to the HindIII site at 45.0 map units (3,918 base pairs). This clearly segregates the VP1 charge phenotype from the cyc phenotype of 3049 in which capsid proteins are overproduced and accumulate in the cytoplasm of infected cells.


Monitoring Arthropods in maize and pasture fields in São Miguel and São Jorge Islands: IPM-Popillia Project.
The dataset presented here is an achievement of the H2020 European project "Integrated Pest Management of the Invasive Japanese Beetle, Popilliajaponica (IPM-Popillia)". This project addresses the challenge of a new risk to plant health in Europe, the invasion of the Japanese beetle, Popilliajaponica (Newman, 1838) (Coleoptera, Rutelidae) and provides an environmentally friendly IPM Toolbox to control the expanding pest populations across Europe. This study aims to present the records of terrestrial arthropod diversity with a special focus on four groups belonging to Carabids and Staphylinid beetles (Coleoptera), Opiliones and Anisolabididae (Dermaptera), collected with the potential to be used as biocontrol agents against P.japonica in future Integrated Pest Management programmes. A thorough sampling programme was conducted in maize and pasture fields in two Islands of the Azores (São Miguel and São Jorge) in the summer of 2022. We provided an inventory of the arthropods recorded in two Azorean agroecosystems (maize and pasture fields) from São Miguel and São Jorge Islands. A total of ten maize and ten pasture fields were sampled and a total of 360 pitfall traps were installed, 216 in São Miguel and 144 in São Jorge, for seven consecutive days in August and September of 2022.We collected 18559 specimens belonging to the phylum Arthropoda, four classes, twelve orders, twenty-six families and forty morphospecies (two identified at the family level as carabid and Staphylinid larvae and 38 identified at the species level). We identified 38 taxa at the species level (n = 18281). Of the 38 identified taxa, 18 species were predators, 15 were plant feeders and five were omnivores. The 18 predators belong to the following families: 10 species were Carabidae, two Staphylinidae, one Anisolabididae, one Chrysopidae, one Leiobunidae, one Nabidae, one Phalangiidae and one Scathophagidae. Concerning the origin of the predators, we recorded five native species: two Carabidae, one Leiobunidae, one Scathophagidae and one Nabidae. The other 13 predator species were introduced or indeterminate.


Insula-based networks in professional musicians: Evidence for increased functional connectivity during resting state fMRI.
Despite considerable research on experience-dependent neuroplasticity in professional musicians, detailed understanding of an involvement of the insula is only now beginning to emerge. We investigated the effects of musical training on intrinsic insula-based connectivity in professional classical musicians relative to nonmusicians using resting-state functional MRI. Following a tripartite scheme of insula subdivisions, coactivation profiles were analyzed for the posterior, ventral anterior, and dorsal anterior insula in both hemispheres. While whole-brain connectivity across all participants confirmed previously reported patterns, between-group comparisons revealed increased insular connectivity in musicians relative to nonmusicians. Coactivated regions encompassed constituents of large-scale networks involved in salience detection (e.g., anterior and middle cingulate cortex), affective processing (e.g., orbitofrontal cortex and temporal pole), and higher order cognition (e.g., dorsolateral prefrontal cortex and the temporoparietal junction), whereas no differences were found for the reversed group contrast. Importantly, these connectivity patterns were stronger in musicians who experienced more years of musical practice, including also sensorimotor regions involved in music performance (M1 hand area, S1, A1, and SMA). We conclude that musical training triggers significant reorganization in insula-based networks, potentially facilitating high-level cognitive and affective functions associated with the fast integration of multisensory information in the context of music performance. Hum Brain Mapp 38:4834-4849, 2017. © 2017 Wiley Periodicals, Inc.


Multi-seasonal water-stress memory versus temperature-driven dynamic structural changes in grapevine.
Perennial plants perpetually adapt to environmental changes in complex and yet insufficiently understood manner. We aimed to separate the intra-seasonal temperature effects on structure and function from perennial and annual water stress effects. This study focused on grapevine (Vitis vinifera L. 'Cabernet Sauvignon') petioles, which being a continuously produced organ, represent the current status of the plant. Field-grown mature plants subjected to multi-annual irrigation treatments (severe water stress, mild water stress and non-stressed) throughout the growing season were compared with greenhouse-grown plants under three temperature regimes (22, 28 and 34 °C). Physiological and functional anatomy parameters were measured. A generalized additive model (GAM) based on meteorological and lysimeter-based field data was applied to determine the relative influence of various meteorological parameters on evapotranspiration (ETc) during the growing season in the field experiment. At the beginning of the growing season, in May, petioles in the severe stress treatment showed a stress-related structure (decreased length, safer hydraulic structure and increased lignification), though having high values of stem water potential (SWP). As the season progressed and temperatures increased, all water availability treatments petioles showed similar changes, and at the end of season, in August, were structurally very similar. Those changes were independent of SWP and were comparable to high temperature-induced changes in the greenhouse. In contrast, stems hydraulic structure was strongly influenced by water availability. Regression analyses indicated a relationship between petioles xylem structure and stomatal conductance (gs), whereas gs (but not SWP) was temperature-dependent. The GAM showed that ETc was mainly dependent on temperature. Our results indicate a perennial water-stress memory response, influencing the petiole structure at the beginning of the following season. Intra-seasonally, the petiole's structure becomes independent of water status, whereas temperature drives the structural changes. Thus, ongoing climate change might disrupt plant performance by purely temperature-induced effects.


Heparin-induced thrombocytopenia after ICD-lead flushing.
Heparin-induced thrombocytopenia (HIT) is a potentially life-threatening prothrombotic complication following heparin administration. We describe a patient, known with idiopathic dilating cardiomyopathy, presenting nine days after a biventricular ICD implantation with dyspnoea and thrombocytopenia. Thirteen days after administration of a single heparin flush during ICD implantation, the patient developed venous thrombosis in two extremities and pulmonary embolism caused by HIT. HIT is the development of thrombocytopenia, caused by IgG antibodies against complexes of platelet factor 4 and heparin, leading to platelet aggregation. HIT may be accompanied by thrombosis in 20-50% of patients and untreated mortality rates are high. Once HIT is suspected, heparin should be replaced by an alternative anti-factor Xa or anti-factor II therapy. Regardless of the low incidence of HIT, because of the widespread use of heparin and the potentially life-threatening course of HIT, all physicians should be aware of it.


Intestinal Pseudo-Obstruction with Life-Threatening Hypokalaemia in a Patient with Adult-Onset Still's Disease.
Adult-onset Still's disease (AOSD) is a rare autoinflammatory disorder that can lead to a cytokine storm, causing a range of symptoms. Acute intestinal pseudo-obstruction is another rare condition that results in intestinal obstruction without anatomical cause. Although the two conditions are rarely reported together, we present the case of a 62-year-old male who developed acute intestinal pseudo-obstruction in the context of an AOSD flare. This led to severe hypokalaemia and a critical condition. Other symptoms included a high-spiking fever lasting for weeks, polyarthralgias and a typical salmon-coloured rash. After ruling out other potential causes, the patient was diagnosed with AOSD. Our findings suggest that the cytokine storm associated with this disease triggered the acute intestinal pseudo-obstruction and life-threatening hypokalaemia, establishing a causal relationship. Only four other cases of AOSD complicated by intestinal pseudo-obstruction have been reported, and this is the first to present with life-threatening hypokalaemia. This case serves as a crucial reminder that, despite being a diagnosis of exclusion, Still's disease should be considered as a potential cause of intestinal pseudo-obstruction, as prompt recognition and treatment of the underlying cause is crucial in managing this potentially life-threatening condition. Acute intestinal pseudo-obstruction is one of the systemic complications that can occur in autoinflammatory diseases such as AOSD, although it has rarely been described.If AOSD presents with abdominal presentation, such as acute intestinal pseudo-obstruction, then amyloidosis should always be carefully considered.


Sustained and transient cells in monkey lateral geniculate nucleus: conduction velocites and response properties.
The receptive-field properties of units in the macaque LGN were studied with extracellular microelectrodes. Analysis of responses of opponent and nonopponent cells to stationary spots, moving spots, and grating revealed transient and sustained varieties of both spectral groups as well as cells with mixed properties. The cell's temporal discharge pattern was not correlated with its color sensitivity. Cells that ere classified as sustained responders did so under all stimulus conditions tested. Most transient cells responded in a transient fashion to each stimulus tested. In a few cells, a sustained response could be shortened into a transient response by activating the cell's surroun mechanism. The conduction velocities of sustained and transient cells were determined by shocking optic chiasm and striate cortex. Among nonopponent cells, transient types conducted significantly faster than the sustained type. No such difference was found among opponent cells. Among all cells, the fastest conducting were transient nonopponent (14 m/s), while the slowest were +B-Y opponent cells (4 m/s). Intermediate were sustained nonopponent (11 m/s) and the remainder of opponent cells types (about 7 m/s). There was a significant positive correlation between a cell's shock-evoked spike latency and its visually evoked latency. These results suggest that there are at least three independent information channels in monkey geniculostriate pathway. Further characterization of these channels is in progress.


Resistance exercise and nutrition to counteract muscle wasting.
Loss of muscle mass is an unfavourable consequence of aging and many chronic diseases. The debilitating effects of muscle loss include declines in physical function and quality of life and increases in morbidity and mortality. Loss of muscle mass is the result of a decrease in muscle protein synthesis, an increase in muscle protein degradation, or a combination of both. Much research on muscle wasting has tended to focus on preventing muscle protein breakdown, and less attention has been paid to providing adequate stimulation to increase muscle protein synthesis. In this review, we present evidence to suggest that interventions aimed at increasing muscle protein synthesis represent the most effective countermeasure for preventing, delaying, or reversing the loss of skeletal muscle mass experienced in various muscle wasting conditions. Based on results from acute and chronic studies in humans in a wide variety of wasting conditions, we propose that resistance exercise training combined with appropriately timed protein (likely leucine-rich) ingestion represents a highly effective means to promote muscle hypertrophy, and may represent a highly effective treatment strategy to counteract the muscle wasting tassociated with aging and chronic disease.


Why states expand Medicaid: party, resources, and history.
After the passage of the Patient Protection and Affordable Care Act in March 2010 and the affirmation of its constitutionality by the Supreme Court in 2012, key decisions about the implementation of health care reform are now in the hands of states. But our understanding of these decisions is hampered by simplistic sortings of state directions into two or three simple, rigid categories. This article takes a different approach--it tracks the variations in relative state progress in implementing Medicaid expansion across a continuum of activities and steps in the decision-making process. This new measure reveals wide variation not only among states that have adopted Medicaid expansion but also among those that have rejected it but have also made progress. We use this new measure to spotlight cross-pressured Republican states that have adopted Medicaid expansion or have prepared to move forward and to explore possible explanations for implementation that extend beyond a simple focus on party control.


Death and Dying in a Digitalized World.
The impact of digitalization on the topic of death and dying seems to be accelerated in recent years. This study aimed to explore the online ways people used to overcome grief and used the COVID-19 restrictions as an example. Thirty-two bereaved participants were interviewed and the data were analyzed using the constructive grounded theory method. Three main themes were extracted from the data: 1) an online way to remember; 2) digitalization of social support, and 3) continuing the bonds. Findings highlighted the important and inevitable role of the digital world in the grief process when there is a restriction in holding usual ceremonies.


Endotoxin protein: a B-cell mitogen and polyclonal activator of C3H/HeJ lymphocytes.
A cell wall protein that is ordinarily complexed to the lipopolysaccharide endotoxin in gram-negative bacteria has been separated by the use of aqueous phenol. The protein is active as a B-cell mitogen and polyclonal activator of murine lymphocytes including the C3H/HeJ strain which is a nonresponder to lipoplysaccharide or lipid A.


[Method of determining the arterial pressure in the collaterals of the extremities by using rheography].
The suggested method permits to record objectively pressure indices in the collaterals of the extremities in case of occlusion of main vessels. The method helps to get more clear idea on the condition of local hemodynamics in patients suffering from vascular diseases and injuries, which is essential for the choice of treatment.


Efficacy and Safety of Denosumab Therapy for Osteogenesis Imperfecta Patients with Osteoporosis-Case Series.
Osteogenesis imperfecta (OI) is a connective tissue disorder that is characterized by low bone density leading to recurrent fractures. The efficacy of the anti-resorption drug denosumab for OI with osteoporosis is still largely unknown. We herein describe the clinical outcomes of eight osteoporotic cases of OI to examine the effects and safety of denosumab. This retrospective, consecutive case series included eight patients respectively aged 42, 40, 14, 22, 3, 51, 37, and 9 years. We measured the bone mineral density (BMD) of the lumbar 1⁻4 spine (L-BMD) and bilateral hips (H-BMD), bone-specific alkaline phosphatase, urinary type I collagen amino-terminal telopeptide, and tartrate-resistant acid phosphatase 5b before and during denosumab therapy. Despite multiple pretreatment fractures in the cohort, no fractures or severe side effects, such as hypocalcemia, were observed during the observational period apart from a fracture in a young pediatric girl. Both L-BMD and H-BMD were increased by denosumab in seven of eight cases. Bone turnover markers were inhibited in most cases by denosumab therapy. Denosumab treatment could generally raise BMD without any adverse effects. The agent therefore represents a good therapeutic option for OI with osteoporosis.


Enhanced susceptibility to end-organ disease in the lupus-facilitating NZW mouse strain.
Although the NZW mouse strain is phenotypically normal, fulminant lupus glomerulonephritis (GN) develops when NZW mice are bred to several other strains, such as NZB, BXSB, B6.Sle1, and B6.Yaa. Based on the observation that aging NZW mice exhibit histologic evidence of GN, we sought to test our hypothesis that NZW mice may be more susceptible to immune-mediated renal damage. NZW mice, as well as C57BL/6 (B6) and BALB/c control mice, were challenged with rabbit anti-glomerular basement membrane nephrotoxic sera (NTS), to induce renal disease. The different mouse strains were monitored for the degree of clinical disease, renal pathology, chemokine profiles, and cellular infiltrates. Although the NZW and control strains showed similar glomerular deposits of rabbit Ig and exhibited similar levels of anti-rabbit xenogeneic immune response, the NZW mice had significantly worse pathologic changes and disease. Compared with the control strains, the NTS-injected NZW mice demonstrated significantly increased proteinuria, elevated blood urea nitrogen levels, more severe histologic GN and tubulointerstitial nephritis, increased glomerular crescent formation with macrophage and neutrophil infiltrates, elevated expression of CC and CXC chemokines (monocyte chemoattractant protein 1, RANTES, KC), and significantly accelerated mortality. Importantly, these changes occurred within a few days after NTS administration. Finally, (B6 x NZW)F(1) mice were as susceptible as the NZW parents, which indicates dominant NZW contributions. Collectively, these findings support the notion that a lupus-facilitating genome may contribute to disease susceptibility by modulating the degree of immune-mediated end-organ damage. The availability of B6-based congenic strains bearing individual NZW-derived lupus susceptibility loci will permit future genetic dissection of end-organ susceptibility in murine lupus.


Effects of vascular-endothelial protein tyrosine phosphatase inhibition on breast cancer vasculature and metastatic progression.
The solid tumor microvasculature is characterized by structural and functional abnormality and mediates several deleterious aspects of tumor behavior. Here we determine the role of vascular endothelial protein tyrosine phosphatase (VE-PTP), which deactivates endothelial cell (EC) Tie-2 receptor tyrosine kinase, thereby impairing maturation of tumor vessels. AKB-9778 is a first-in-class VE-PTP inhibitor. We examined its effects on ECs in vitro and on embryonic angiogenesis in vivo using zebrafish assays. We studied the impact of AKB-9778 therapy on the tumor vasculature, tumor growth, and metastatic progression using orthotopic models of murine mammary carcinoma as well as spontaneous and experimental metastasis models. Finally, we used endothelial nitric oxide synthase (eNOS)-deficient mice to establish the role of eNOS in mediating the effects of VE-PTP inhibition. All statistical tests were two-sided. AKB-9778 induced ligand-independent Tie-2 activation in ECs and impaired embryonic zebrafish angiogenesis. AKB-9778 delayed the early phase of mammary tumor growth by maintaining vascular maturity (P < .01, t test); slowed growth of micrometastases (P < .01, χ(2) test) by preventing extravasation of tumor cells (P < 0.01, Fisher exact test), resulting in a trend toward prolonged survival (27.0 vs 36.5 days; hazard ratio of death = 0.33, 95% confidence interval = 0.11 to 1.03; P = .05, Mantel-Cox test); and stabilized established primary tumor blood vessels, enhancing tumor perfusion (P = .03 for 4T1 tumor model and 0.05 for E0771 tumor model, by two-sided t tests) and, hence, radiation response (P < .01, analysis of variance; n = 7 mice per group). The effects of AKB-9778 on tumor vessels were mediated in part by endothelial nitric oxide synthase activation. Our results demonstrate that pharmacological VE-PTP inhibition can normalize the structure and function of tumor vessels through Tie-2 activation, which delays tumor growth, slows metastatic progression, and enhances response to concomitant cytotoxic treatments.


Thickness of Saturn's Rings Inferred from Voyager 1 Observations of Microwave Scatter.
Earth-based telescopic observations indicate that Saturn's rings are about 1 kilometer thick, while spacecraft measurements and theoretical considerations give an upper bound of about 100 meters. Analysis of a shielding effect present in radio occultation provides a sensitive new measure of the ring thickness. On the basis of this effect, Voyager 1 microwave measurements of near-forward scatter imply a thickness ranging from less than 10 meters in ring C to about 20 and 50 meters in the Cassini division and ring A, respectively. Monolayer models do not fit the observations in the latter two regions. The discrepancy between the Earth-based and spacecraft measurements may be due to warps in the ring plane or effects of tenuous material outside the primary ring system.


"The importance of awareness, support and inner strength to balance everyday life" - a qualitative study about women's experiences of a workplace health promotion program in human service organizations in Sweden.
In many European countries, women have a higher sickness absence rate than men. Women also report higher levels of work-life conflict, which has a negative impact on women's self-perceived health. Interventions studies on work-life balance literature are scarce. This research adds knowledge about work-life balance by examining female employees' experiences of a newly developed intervention program, the BELE program (Balance in Everyday Life Empowerment program), aiming to enhance the work-life balance and wellbeing of female employees. All participants in the BELE program were included in the sample (n = 55) of the study. The methods used for gathering data were focus group interviews (n = 8) and semi-structured individual interviews (n = 8). A qualitative content analysis was used for the analyses. The results showed that a majority of the participants experienced a process of change for enhancement of wellbeing and balance in their everyday life when taking part in the BELE program. In the analyses, three main themes emerged. Reflecting and strengthening inner resource, Trying to change everyday patterns and habits and Experiencing more balance and well-being. The results point out the importance of awareness and reflection on everyday life to promote work-life balance and wellbeing. The BELE program was described as a wakeup call to one's own life and as an enhancer of empowerment processes and equality in the homes. Moreover, the results showed the need for health education in the workplace focusing on both work and private life to enhance balance and wellbeing among female employees'. The findings indicate that it is important to work at individual and group levels in work-life balance interventions and not merely at the organizational level or in the workplace setting.


Induction of apoptosis and inhibition of small cell lung cancer growth by the quinoxaline tyrphostins.
Coexpression of the Kit receptor tyrosine kinase and its ligand, stem cell factor (SCF), occurs in a high proportion of small cell lung cancers (SCLCs) and drives an autocrine loop that enhances proliferation. To determine whether this autocrine loop affects apoptosis, SCLC cells expressing only SCF or both SCF and Kit were deprived of growth factors for 72 h and the relative number of cells undergoing apoptosis was assessed using nuclear DNA content and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assays. Coexpression of SCF and Kit inhibited apoptosis; apoptosis could, in turn, be enhanced by the addition of the quinoxaline tyrosine kinase inhibitors, which are specific antagonists of the platelet-derived growth factor receptor and Kit. Treatment of the H526 cell line, which is growth-stimulated by soluble SCF, with AG1296 resulted in a marked decrease in growth and an increase in apoptosis in a dose-dependent fashion. Growth inhibition correlated well with the inhibition of Kit tyrosine phosphorylation. The AG1296 compound at its maximum soluble concentration inhibited the growth of 5 of 6 SCLC cell lines in complete medium by an average of 50%. These data suggest that optimized pharmacological inhibitors of Kit activity may be a new class of compounds potentially useful in the treatment of SCLC.


Spatiotemporal Trends Spanning Three Decades Show Toxic Levels of Chemical Contaminants in Marine Mammals.
Despite their ban and restriction under the 2001 Stockholm Convention, persistent organic pollutants (POPs) are still widespread and pervasive in the environment. Releases of these toxic and bioaccumulative chemicals are ongoing, and their contribution to population declines of marine mammals is of global concern. To safeguard their survival, it is of paramount importance to understand the effectiveness of mitigation measures. Using one of the world's largest marine mammals strandings data sets, we combine published and unpublished data to examine pollutant concentrations in 11 species that stranded along the coast of Great Britain to quantify spatiotemporal trends over three decades and identify species and regions where pollutants pose the greatest threat. We find that although levels of pollutants have decreased overall, there is significant spatial and taxonomic heterogeneity such that pollutants remain a threat to biodiversity in several species and regions. Of individuals sampled within the most recent five years (2014-2018), 48% of individuals exhibited a concentration known to exceed toxic thresholds. Notably, pollutant concentrations are highest in long-lived, apex odontocetes (e.g., killer whales (Orcinus orca), bottlenose dolphins (Tursiops truncatus), and white-beaked dolphins (Lagenorhynchus albirostris)) and were significantly higher in animals that stranded on more industrialized coastlines. At the present concentrations, POPs are likely to be significantly impacting marine mammal health. We conclude that more effective international elimination and mitigation strategies are urgently needed to address this critical issue for the global ocean health.


Circulating mediators of inflammation and immune activation in AIDS-related non-hodgkin lymphoma.
Non-Hodgkin lymphoma (NHL) is the most common AIDS-related malignancy in developed countries. An elevated risk of developing NHL persists among HIV-infected individuals in comparison to the general population despite the advent of effective antiretroviral therapy. The mechanisms underlying the development of AIDS-related NHL (A-NHL) are not fully understood, but likely involve persistent B-cell activation and inflammation. This was a nested case-control study within the ongoing prospective Multicenter AIDS Cohort Study (MACS). Cases included 47 HIV-positive male subjects diagnosed with high-grade B-cell NHL. Controls were matched to each case from among participating HIV-positive males who did not develop any malignancy. Matching criteria included time HIV+ or since AIDS diagnosis, age, race and CD4+ cell count. Sera were tested for 161 serum biomarkers using multiplexed bead-based immunoassays. A subset of 17 biomarkers, including cytokines, chemokines, acute phase proteins, tissue remodeling agents and bone metabolic mediators was identified to be significantly altered in A-NHL cases in comparison to controls. Many of the biomarkers included in this subset were positively correlated with HIV viral load. A pathway analysis of our results revealed an extensive network of interactions between current and previously identified biomarkers. These findings support the current hypothesis that A-NHL develops in the context of persistent immune stimulation and inflammation. Further analysis of the biomarkers identified in this report should enhance our ability to diagnose, monitor and treat this disease.


Developmental changes of chicken liver AMP deaminase.
The AMP deaminase activity measured in crude chicken liver extract did not change significantly during development. The livers of 10- and 14-day chick embryos, 1-day, 5-, 10- and 16-week-old chickens and adult hens were examined for the existence of multiple forms of AMP deaminase. Phosphocellulose column chromatography revealed the existence of two peaks of enzyme activity in the liver of 10- and 16-week-old chickens and adult hens. Kinetic studies with the preparations of AMP deaminase revealed sigmoid-shaped substrate-saturation curves at all developmental stages and hyperbolic-shaped saturation curves for the enzyme form appearing in 10-week-old chickens. All AMP deaminases investigated were susceptible to activation by ATP and inhibition by Pi. Kinetic and regulatory properties as well as pH optima of all the enzyme preparations tested indicate that AMP deaminase isolated from the embryos and from 1-day-old chicks was similar to the form I isolated from adult hens and differed significantly from the form II of this enzyme.


Thermodynamic studies on the solvent effects in chromatography on molecularly imprinted polymers. 1. Nature of the organic modifier.
Molecularly imprinted polymers (MIPs) are used as highly enantioselective stationary phases in liquid chromatography. To optimize the binding performance of MIPs, different types of polar modifiers are frequently used. Previous studies have shown that the hydrogen-bonding donor parameter (HBD) of the modifier has a large influence on the binding performance of MIPs in chiral separations. This possibility is addressed in a detailed thermodynamic study of a Fmoc-L-tryptophan (Fmoc-L-Trp) imprinted polymer, eluted with four different polar modifiers, i.e., THF, propan-2-ol, methanol, and acetic acid, which have different HBDs (0.00, 0.33, 0.43, and 0.61, respectively). Adsorption isotherm data for each enantiomer in each of these organic modifiers were acquired by frontal analysis over a 20 000 dynamic concentration range. Nonlinear regression of the isotherm data, along with independent calculation of the affinity energy distributions, identified four different types of binding sites coexisting for the enantiomers on the MIP. The exception was acetic acid, which has the highest HBD. In this case, three types of binding sites only coexist on the MIP. The isotherm parameters obtained from these data show the following: (1) The association energies of the two enantiomers with a given type of sites have a similar magnitude; however, the density of the sites is higher for the template than for its antipode. (2) The nature of the organic modifier has a larger influence on the density of high-energy sites than on the association constant of these sites. (3) The molecular size of the organic modifier has a larger influence on the site density (especially for Fmoc-D-Trp) than does HBD. (4) Using an organic modifier with a higher HBD reduces the enantioselectivity on each site. (5) High-energy sites are more enantioselective than low-energy ones. (6) Using an organic modifier with a high HBD causes a larger reduction in the density of high-energy sites approached by the template molecules.


JAM-B regulates maintenance of hematopoietic stem cells in the bone marrow.
In adult mammals, hematopoietic stem cells (HSCs) reside in the bone marrow (BM) and are maintained in a quiescent and undifferentiated state through adhesive interactions with specialized microenvironmental niches. Although junctional adhesion molecule-C (JAM-C) is expressed by HSCs, its function in adult hematopoiesis remains elusive. Here, we show that HSCs adhere to JAM-B expressed by BM stromal cells in a JAM-C dependent manner. The interaction regulates the interplay between HSCs and BM stromal cells as illustrated by the decreased pool of quiescent HSCs observed in jam-b deficient mice. We further show that this is probably because of alterations of BM stromal compartments and changes in SDF-1α BM content in jam-b(-/-) mice, suggesting that JAM-B is an active player in the maintenance of the BM stromal microenvironment.


Biophysical characterization of KV3.1 potassium channel activating compounds.
The effect of two positive modulators, RE1 and EX15, on the voltage-gated K(+) channel Kv3.1 was investigated using the whole-cell patch-clamp technique on HEK293 cells expressing Kv3.1a. RE1 and EX15 increased the Kv3.1 currents in a concentration-dependent manner with an EC50 value of 4.5 and 1.3µM, respectively. However, high compound concentrations caused an inhibition of the Kv3.1 current. The compound-induced activation of Kv3.1 channels showed a profound hyperpolarized shift in activation kinetics. 30µM RE1 shifted V1/2 from 5.63±0.31mV to -9.71±1.00mV and 10µM EX15 induced a shift from 10.77±0.32mV to -15.11±1.57mV. The activation time constant (Tauact) was reduced for both RE1 and EX15, with RE1 being the fastest activator. The deactivation time constant (Taudeact) was also markedly reduced for both RE1 and EX15, with EX15 inducing the most prominent effect. Furthermore, subjected to depolarizing pulses at 30Hz, both compounds were showing a use-dependent effect resulting in a reduction of the compound-mediated effect. However, during these conditions, RE1- and EX15-modified current amplitudes still exceeded the control condition amplitudes by up to 200%. In summary, the present study introduces the first detailed biophysical characterization of two new Kv3.1 channel modifying compounds with different modulating properties.


The hemibiotrophic cacao pathogen Moniliophthora perniciosa depends on a mitochondrial alternative oxidase for biotrophic development.
The tropical pathogen Moniliophthora perniciosa causes witches' broom disease in cacao. As a hemibiotrophic fungus, it initially colonizes the living host tissues (biotrophic phase), and later grows over the dead plant (necrotrophic phase). Little is known about the mechanisms that promote these distinct fungal phases or mediate the transition between them. An alternative oxidase gene (Mp-aox) was identified in the M. perniciosa genome and its expression was analyzed througout the fungal life cycle. In addition, the effects of inhibitors of the cytochrome-dependent respiratory chain (CRC) and alternative oxidase (AOX) were evaluated on the in vitro development of M. perniciosa. Larger numbers of Mp-aox transcripts were observed in the biotrophic hyphae, which accordingly showed elevated sensitivity to AOX inhibitors. More importantly, the inhibition of CRC prevented the transition from the biotrophic to the necrotrophic phase, and the combined use of a CRC and AOX inhibitor completely halted fungal growth. On the basis of these results, a novel mechanism is presented in which AOX plays a role in the biotrophic development of M. perniciosa and regulates the transition to its necrotrophic stage. Strikingly, this model correlates well with the infection strategy of animal pathogens, particularly Trypanosoma brucei, which uses AOX as a strategy for pathogenicity.


A randomized effectiveness trial of cognitive-behavioral therapy and medication for primary care panic disorder.
Panic disorder is a prevalent, often disabling condition among patients in the primary care setting. Although numerous studies have assessed the effectiveness of treatments for depression in primary care, few such studies have been conducted for panic disorder. To implement and test the effectiveness of a combined pharmacotherapy and cognitive-behavioral intervention for panic disorder tailored to the primary care setting. Randomized, controlled study comparing intervention to treatment as usual. Six primary care clinics associated with 3 university medical schools, serving an ethnically and socioeconomically diverse patient population. Two hundred thirty-two primary care patients meeting DSM-IV criteria for panic disorder. Comorbid mental and physical disorders were permitted, provided these did not contraindicate the treatment to be provided and were not acutely life threatening. Patients were randomized to receive either treatment as usual or an intervention consisting of a combination of up to 6 sessions (across 12 weeks) of cognitive-behavioral therapy (CBT) modified for the primary care setting, with up to 6 follow-up telephone contacts during the next 9 months, and algorithm-based pharmacotherapy provided by the primary care physician with guidance from a psychiatrist. Behavioral health specialists, the majority inexperienced in CBT for panic disorder, were trained to deliver the CBT and coordinated overall care, including pharmacotherapy. Proportion of subjects remitted (no panic attacks in the past month, minimal anticipatory anxiety, and agoraphobia subscale score <10 on Fear Questionnaire) and responding (Anxiety Sensitivity Index score <20) and change over time in World Health Organization Disability Scale and short form 12 scores. The combined cognitive-behavioral and pharmacotherapeutic intervention resulted in sustained and gradually increasing improvement relative to treatment as usual, with significantly higher rates at all points of both the proportion of subjects remitted (3 months, 20% vs 12%; 12 months, 29% vs 16%) and responding (3 months, 46% vs 27%; 12 months, 63% vs 38%) and significantly greater improvements in World Health Organization Disability Scale (all points) and short form 12 mental health functioning (3 and 6 months) scores. These effects were obtained in spite of similar rates of delivery of guideline-concordant pharmacotherapy to the 2 groups. Delivery of evidence-based CBT and medication using the collaborative care model and a CBT-naive, midlevel behavioral health specialist is feasible and significantly more effective than usual care for primary care panic disorder.


Restless legs syndrome as side effect of second generation antidepressants.
Although of clinical interest, the question is still not fully answered whether antidepressants (AD) can cause or exacerbate restless legs syndrome (RLS). The literature provides contradictory information. This study addresses this problem for the class of second generation AD. In four neurological offices, all patients treated for the first time with an AD were prospectively observed with regard to the question of whether RLS occurred or pre-existing RLS worsened as a result of the medication. Because initial treatment in the participating offices is mainly executed with "modern" selective AD, the study was restricted to these drugs (fluoxetine, paroxetine, citalopram, sertraline, escitalopram, venlafaxine, duloxetine, reboxetine, and mirtazapine). In 9% of patients, RLS was recorded as a side effect related to the administration of AD. The frequency of this side effect varied among the drugs. The problem is most pronounced with mirtazapine provoking or deteriorating RLS in 28% of patients. By contrast, no case occurred during use of reboxetine. As for the other AD, the rate of newly occurred and deteriorated RLS, ranged from 5% to 10%. Typically, RLS occurred during the initial days of treatment.


The lightness of water vapor helps to stabilize tropical climate.
Moist air is lighter than dry air at the same temperature, pressure, and volume because the molecular weight of water is less than that of dry air. We call this the vapor buoyancy effect. Although this effect is well documented, its impact on Earth's climate has been overlooked. Here, we show that the lightness of water vapor helps to stabilize tropical climate by increasing the outgoing longwave radiation (OLR). In the tropical atmosphere, buoyancy is horizontally uniform. Then, the vapor buoyancy in the moist regions must be balanced by warmer temperatures in the dry regions of the tropical atmosphere. These higher temperatures increase tropical OLR. This radiative effect increases with warming, leading to a negative climate feedback. At a near present-day surface temperature, vapor buoyancy is responsible for a radiative effect of 1 W/m2 and a negative climate feedback of about 0.15 W/m2 per kelvin.


Increased protein synthesis after T cell activation in presence of cyclosporin A.
The immunosuppressive drug, cyclosporin A (CsA), blocks immune responses by inhibiting the calcineurin-dependent dephosphorylation of the nuclear factor of activated T cells (NFAT). We have previously reported that T cells activated in presence of CsA exhibit particular properties. In our study, we have tested the hypothesis that T cells activated in presence of CsA display a differential pattern of gene expression. T lymphocytes were activated in vitro by Concanavalin A with or without CsA. The cells were: (1) pulsed with 35S-methionine to label the newly synthesized proteins that in turn were revealed by 2D-gel electrophoresis; (2) analyzed by flow cytometry for activation markers expression; and (3) examined by gel electrophoresis for early tyrosine phosphorylation events. The proteomic patterns of T lymphocytes activated by Concanavalin A, with or without CsA, were compared. In keeping with the well-known effect of the immunosuppressor, many polypeptides were not found in its presence. Remarkably, several newly synthesized polypeptides were detected only when activation was carried out in presence of CsA. In addition, immunologically relevant proteins, such as CD44 and CD69, escape CsA-inhibitory action. Furthermore, CsA did not modify the early protein tyrosine phosphorylation events resulting from T cell triggering. The present data show that the effect of CsA on protein synthesis is more complex than anticipated. Signaling provided by T cell activation and the blockade of the calcineurin-dependent pathway by CsA results in an altered program of gene expression.


[Nosoareas of clonorchiasis in the Primorye Territory].
The mammalian liver parasite (Clonorchis sinensis flukes) has spread in the Primorye Territory. The areas of the first intermediate hosts for the parasite mollusks of the genus Parafossarulus have been found to recently become wider south-western predominantly due to lotus acclimatization, resulting in the expansion of a parasite habitation area. The intensity of newly formed foci of clonorchiasis is an order of magnitude greater than that of natural ones and they are a higher hazard since they have formed in the recreation areas used by the population for rest and fishing. The carp is the second intermediate host of trematodes. Patients' medical history data show that European and crucian carps (less frequently skygazer, rudd, gudgeon, minnow) have served as the main sources of human infection with trematodes.


So you want to express your protein in Escherichia coli?
Recombinant proteins have been extensively employed as therapeutics for the treatment of various critical and life-threatening diseases and as industrial enzymes in high-value industrial processes. Advances in genetic engineering and synthetic biology have broadened the horizon of heterologous protein production using multiple expression platforms. Selection of a suitable expression system depends on a variety of factors ranging from the physicochemical properties of the target protein to economic considerations. For more than 40 years, Escherichia coli has been an established organism of choice for protein production. This review aims to provide a stepwise approach for any researcher embarking on the journey of recombinant protein production in E. coli. We present an overview of the challenges associated with heterologous protein expression, fundamental considerations connected to the protein of interest (POI) and designing expression constructs, as well as insights into recently developed technologies that have contributed to this ever-growing field.


The proton pump interactor (Ppi) gene family of Arabidopsis thaliana: expression pattern of Ppi1 and characterisation of knockout mutants for Ppi1 and 2.
Plant plasma membrane H+-ATPases (PM H+-ATPase) are essential for establishing a proton electrochemical gradient across the cell plasma membrane. Their regulation is poorly understood, except for the role of 14-3-3 proteins, which relieve autoinhibition from the C-terminal domain. A novel protein interacting with this domain was recently identified in Arabidopsis and named PPI1 (Proton Pump Interactor 1). PPI1 stimulates PM H+-ATPase activity in vitro. Here, we analyse the expression pattern of Ppi1 using beta-glucuronidase as a reporter. Expression is strong in root and shoot vascular systems, particularly in meristematic and sink tissues, as well as in pollen, stigmas and siliques, but not in developing embryos. Removal of the first intron decreased GUS expression 45-fold. We also analysed the transcription of Ppi2, another gene in the family, and demonstrated that Ppi2 is expressed in seedlings, cultured cells and flowers. We reassessed Ppi2 gene structure based on RT-PCR amplifications, cDNA data and similarity to other Ppi genes. Insertional mutants for both Ppi1 and Ppi2 were isolated. Two different mutants of Ppi1 showed aberrant mRNAs and lacked any detectable protein and are therefore true knockouts. Interestingly, one mutation inhibited the splicing of one intron at a considerable distance (>700 bp) from the T-DNA insertion site, but not the splicing of a proximal intron (29 bp) or of any other intron. At the plant level, neither of the single mutants nor the double ppi1ppi2 mutant showed an altered phenotype in standard growth conditions under acid load or salt stress.


Endoscopic ultrasound appearance of pancreatic serotonin-staining neuroendocrine neoplasms.
The pancreatic localization of serotonin-staining neuroendocrine neoplasms is extremely rare. This is a retrospective study aimed at analyzing the endoscopic ultrasound appearance of pancreatic serotoninoma. Between 2010 and 2016, all consecutive patients with histologically proven pancreatic serotoninoma who had undergone endoscopic ultrasound were enrolled. Eight patients (six F, median age 68.5 years) had a diagnosis of pancreatic serotoninoma and underwent endoscopic ultrasound examinations. Median diameter of the lesion was ten mm. The nodule echotexture was hypoechoic in seven out of eight cases. The most frequent localization was the pancreatic neck (four); in three cases, the tumor was located in the pancreatic head and in one in the body. In seven cases the tumor caused a main pancreatic duct dilation; in three cases also the secondary ducts were dilated. In one case a dilation of the common bile duct was observed. At contrast-enhanced endoscopic ultrasound no one showed the typical contrast-enhancement. Elastography (available in two patients) showed a rigid pattern of the lesion. From this case series a specific endoscopic ultrasound appearance resulted for pancreatic serotoninoma, different from other types of pancreatic neuroendocrine neoplasm, but it is difficult to differentiate it from a pancreatic adenocarcinoma or an intraductal papillary mucinous neoplasm.


The dimeric nature of bonding in gallium: from small clusters to the α-gallium phase.
We consider the structural similarity of small gallium clusters to the bulk structure of α-gallium, which has been previously described as a molecular metal, via density functional theory-based computations. Previous calculations have shown that the tetramer, the hexamer, and the octamer of gallium are all structurally similar to the α-phase. We perform an analysis of the bonding in these clusters in terms of the molecular orbitals and atoms in molecules description in order to assess whether we can see similarities at these sizes to the bonding pattern, which is ascribed to the co-existence of covalent and metallic bonding in the bulk. The singlet Ga4 and Ga8 clusters can be constructed in a singlet ground state from the Ga-dimers in the first excited triplet state of the Ga2-molecule, the (3)Σg(-) state. Molecular orbital (MO) analysis confirms that the dimer is an essential building block of these small clusters. Comparison of the AIM characteristics of the bonds within the clusters to the bonds in the bulk α-phase supports the identification of the covalent bond in the bulk as related to the (3)Σg(-) state of the dimer.


Safety profile of ultrasmall superparamagnetic iron oxide ferumoxtran-10: phase II clinical trial data.
The safety data from the phase II clinical trial of ferumoxtran-10, an ultrasmall superparamagnetic iron oxide contrast agent, are presented. One hundred and four patients with focal liver or spleen pathologies underwent ferumoxtran-10-enhanced magnetic resonance (MR) imaging at doses of 0.8, 1.1, and 1.7 mg Fe/kg. Overall, 15% patients reported a total of 33 adverse events, regardless of causality. The adverse events most frequently seen were dyspnea (3.8%), chest pain (2.9%), and rash (2.9%). No serious adverse events were reported during the 48 hour observation period. There were no clinically significant effects on vital signs, physical examination, and laboratory results. Ferumoxtran-10 is a safe and well tolerated MR contrast agent.


Direct effects of hepatitis C virus on the lymphoid cells.
It has been reported that the direct binding of hepatitis C virus (HCV) and/or the replication of HCV in the extrahepatic organs and, especially, lymphoid cells, might affect the pathogenesis of extrahepatic diseases with HCV infection. More than one decade ago, several reports described the existence of HCV-RNA in peripheral blood mononuclear cells. Moreover, many reports describing the existence of HCV in B lymphocytes and B cell lymphoma have been published. In addition to B lymphocytes, it was reported that HCV replication could be detected in T lymphocytes and T cell lines. Among the extrahepatic diseases with HCV infection, mixed cryoglobulinemia-related diseases and autoimmune-related diseases are important for understanding the immunopathogensis of HCV persistent infection. Moreover, HCV persistent infection can cause malignant lymphoma. The biological significance of lymphotropic HCV has not yet become clear. However, several candidates have been considered for a long time. One is that lymphotropic HCV is an HCV reservoir that might contribute to the recurrence of HCV infection and difficult-to-treat disease status. The other important issue is the carcinogenesis of the lymphoid cells and disturbances of the immune responses. Therefore, the extrahepatic diseases might be induced by direct interaction between HCV and lymphoid cells. In this article, we summarize various studies showing the direct effect of HCV on lymphoid cells and discuss the biological significance of lymphotropic HCV.


Cutting Edge: TFII-I controls B cell proliferation via regulating NF-kappaB.
The multifunctional transcription factor TFII-I physically and functionally interacts with Bruton's tyrosine kinase in murine B cells. However, the downstream functions of TFII-I in B cells are unknown. Toward achieving this goal, we established stable posttranscriptional silencing of TFII-I in WEHI-231 immature murine B cells, which undergoes growth arrest and apoptosis either upon anti-IgM or TGF-beta signaling. In this study, we show that TFII-I promotes growth arrest of cells in a signal-dependent manner. Unlike control cells, B cells exhibiting loss of TFII-I function fail to undergo arrest upon signaling due to up-regulation of c-Myc expression and concomitant down-regulation of both p21 and p27. Loss of TFII-I is also associated with simultaneous increase in nuclear c-rel and decrease in p50 homodimer binding. Thus, besides controlling c-myc transcription, TFII-I controls B cell proliferation by regulating both nuclear translocation of c-rel and DNA-binding activity of p50 NF-kappaB.


Muscarinic M1 and M4 receptor agonists for schizophrenia: promising candidates for the therapeutic arsenal.
Successful phase 3 trials of KarXT in people with schizophrenia herald a new era of treating the disorder with drugs that do not target the dopamine D2 receptor. The active component of KarXT is xanomeline, a muscarinic (CHRM) M1 and M4 agonist, making muscarinic receptors a viable target for treating schizophrenia. This review covers the process of taking drugs that activate the muscarinic M1 and M4 receptors from conceptualization to the clinic and details the mechanisms by which activating the CHRM1 and 4 can affect the broad spectrum of symptoms experienced by people with schizophrenia. Schizophrenia is a syndrome which means drugs that activate muscarinic M1 and M4 receptors, as was the case for antipsychotic drugs acting on the dopamine D2 receptor, will not give optimal outcomes in everyone within the syndrome. Thus, it would be ideal to identify people who are responsive to drugs activating the CHRM1 and 4. Given knowledge of the actions of these receptors, it is possible treatment non-response could be restricted to sub-groups within the syndrome who have deficits in cortical CHRM1 or those with one of the cognitive endophenotypes that may be identifiable by changes in the blood transcriptome.


National British Orthodontic Society (BOS) Orthognathic Audit 2017-2018.
To carry out a UK national clinical audit of orthognathic acceptance criteria and information provided to orthognathic patients before treatment. National clinical audit. Data collected using Bristol Online Surveys. Sixty-nine UK hospital orthodontic departments submitted data. Data were collected at two time points using Bristol Online Surveys over a period of 12 months. These were before treatment at the first multidisciplinary clinic (MDT) and immediately after surgery. The data collected included: Index of Orthognathic Functional Treatment Need (IOFTN); Index of Orthodontic Treatment Need (IOTN); age; previous orthodontic treatment; attendance at an MDT; treatment times; and information provision. Eighty-five units agreed to take part in the audit with 69 submitting data, giving a response rate of 81%. The data from 3404 patients were uploaded, 2263 before treatment and 1141 immediately after surgery. Of patients, 91.07% had an IOFTN score of 4 or 5 and 88.73% had an IOTN score of 4 or 5. The mean age at the first MDT was 22 years in the first cohort and 21 years and 4 months in the second immediate post-surgery cohort. Of patients, 37.93% had undergone some form of previous orthodontic treatment, but only 0.28% had undergone previous orthognathic treatment; 96.93% had an MDT confirm that orthodontic treatment by itself was insufficient to adequately correct their functional symptoms. The average treatment time from bond up to surgery was 2 years and 6 months. With respect to information provision, patients received information from a number of sources, principally the British Orthodontic Society (BOS) patient information leaflets and the BOS website Your Jaw Surgery. In the UK, the majority of orthognathic cases fulfil the criteria for acceptance for NHS-funded orthognathic treatment, as outlined by the Chief Dental Officer's interim guidance on orthognathic treatment. This suggests any prior approval process would not be a good use of NHS resources in the commissioning of orthognathic treatment.


Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts.
The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. When expressed in Chinese hamster ovary cells this was glycosylated with a combination of high mannose and complex chains. The latter chains were shown by chromatographic separation and mass spectrometric analysis of tryptic digests to be clustered in the EGF-binding domain. Treatment with the endoglycosidase, peptide-N-glycosidase F (PNGase F), reduced the molecular weight from 110 kDa to 75 kDa. Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. The data were consistent with the complex chains being trisialylated tetra-antennary oligosaccharides fucosylated on the reducing terminal GlcNAc. The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4.


Presenting features of idiopathic versus secondary restless legs syndrome in pregnancy.
Restless legs syndrome (RLS) is the most common movement disorder in pregnancy, which can be idiopathic or secondary. There are limited comparative data regarding these two forms of RLS. The aim of this study was to compare clinical features of idiopathic and secondary RLS in pregnant women. Over a period of 3 months, 443 women who admitted for delivery in two clinical centers were screened for RLS using four diagnostic criteria of the international RLS study group. A total of 79 subjects diagnosed with RLS were consecutively enrolled in the present study. All of them were interviewed for medical history and complaints during pregnancy and responded to self-administer international RLS rating scale. Ten subjects (12.9%) out of 79 pregnant women with RLS had idiopathic form, and their mean age was significantly higher than patients with secondary RLS (30.6 ± 7.3 years vs. 26.4 ± 4.6 years, P = 0.0260). Compared with women with secondary RLS, sleep duration in pregnancy was significantly decreased in idiopathic RLS group (P = 0.0460), whereas RLS severity score was similar in both groups. No significant difference was observed between the two groups in terms of other sleep complaints, the positive family history of RLS, parity, duration of pregnancy, or frequency of cesarean section (P > 0.0500). Idiopathic and secondary RLS have relatively similar courses and features during pregnancy. However, the idiopathic form may have more negative impact on sleep in pregnancy. Careful screening and effective treatment of idiopathic RLS before pregnancy is recommended to limit these disturbances.


The risk of hydroquinone and sunscreen over-absorption via photodamaged skin is not greater in senescent skin as compared to young skin: nude mouse as an animal model.
Intrinsic aging and photoaging modify skin structure and components, which subsequently change percutaneous absorption of topically applied permeants. The purpose of this study was to systematically evaluate drug/sunscreen permeation via young and senescent skin irradiated by ultraviolet (UV) light. Both young and senescent nude mice were subjected to UVA (10 J/cm(2)) and/or UVB radiation (175 mJ/cm(2)). Physiological parameters, immunohistology, and immunoblotting were employed to examine the aged skin. Hydroquinone and sunscreen permeation was determined by in vitro Franz cell. In vivo skin absorption was documented using a hydrophilic dye, rhodamine 123 (log P=-0.4), as a permeant. UVA exposure induced cyclooxygenase (COX)-2 and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) upregulation. Epidermal tight junction (TJ) were degraded by UVA. UVB increased transepidermal water loss (TEWL) from 13 to 24 g/m(2)/h. Hyperplasia and inflammation, but not loss of TJ, were also observed in UVB-treated skin. UVA+UVB- and UVA-irradiated skin demonstrated similar changes in histology and biomarkers. UVA+UVB or UVA exposure increased hydroquinone flux five-fold. A negligible alteration of hydroquinone permeation was shown with UVB exposure. Hydroquinone exhibited a lower penetration through senescent skin than young skin. Both UVA and UVB produced enhancement of oxybenzone flux and skin uptake. However, the amount of increase was less than that of hydroquinone delivery. Photoaging did not augment skin absorption of sunscreens with higher lipophilicity, including avobenzone and ZnO. Exposure to UVA generally increased follicular entrance of these permeants, which showed two- to three-fold greater follicular uptake compared to the untreated group. Photoaging had less impact on drug/sunscreen absorption with more lipophilic permeants. Percutaneous absorption did not increase in skin subjected to both intrinsic and extrinsic aging.


Cardiac Evaluation of Children With a Family History of Sudden Death.
After sudden death occurs in the young, first-degree family members should undergo clinical screening for occult cardiac disease, but the diagnostic yield from screening is not well-defined in the United States. The purpose of this study was to determine the clinical predictors of cardiac diagnosis in children referred for evaluation following a sudden death in the family. Patients referred for a family history of sudden death were evaluated in a retrospective review from a tertiary pediatric referral center. Among 419 pediatric relatives of 256 decedents, 27% of patients were diagnosed with a disease or had a clinical finding of uncertain significance. Patients were diagnosed with heritable cardiac disease in 39 cases (9.3%). Nonheritable cardiac disease was diagnosed in another 5.5% of patients. Clinical findings of uncertain significance were present in 52 patients (12.4%), including abnormal electrophysiological test results (41 of 52) or imaging test results (11 of 52). Among patients diagnosed with a heritable cardiac disease, the nearest affected relative was almost always a first-degree relative (37 of 39, 95%). The strongest predictors for a successful diagnosis in the patient were an abnormal electrocardiogram and a first-degree relationship to the nearest affected relative (odds ratios: 24.2 and 18.8, respectively). Children referred for a family history of sudden death receive cardiac disease diagnoses (14%), but clinical findings of uncertain significance increase the challenge of clinical management. The importance of a diagnosis in first-degree affected relatives supports the clinical practice of testing intervening family members first when patients are second- or higher-degree relatives to the decedent.