The complete plastid genomes of Ophrys iricolor and O. sphegodes (Orchidaceae) and comparative analyses with other orchids.
Sexually deceptive orchids of the genus Ophrys may rapidly evolve by adaptation to pollinators. However, understanding of the genetic basis of potential changes and patterns of relationships is hampered by a lack of genomic information. We report the complete plastid genome sequences of Ophrys iricolor and O. sphegodes, representing the two most species-rich lineages of the genus Ophrys. Both plastomes are circular DNA molecules (146754 bp for O. sphegodes and 150177 bp for O. iricolor) with the typical quadripartite structure of plastid genomes and within the average size of photosynthetic orchids. 213 Simple Sequence Repeats (SSRs) (31.5% polymorphic between O. iricolor and O. sphegodes) were identified, with homopolymers and dipolymers as the most common repeat types. SSRs were mainly located in intergenic regions but SSRs located in coding regions were also found, mainly in ycf1 and rpoC2 genes. The Ophrys plastome is predicted to encode 107 distinct genes, 17 of which are completely duplicated in the Inverted Repeat regions. 83 and 87 putative RNA editing sites were detected in 25 plastid genes of the two Ophrys species, all occurring in the first or second codon position. Comparing the rate of nonsynonymous (dN) and synonymous (dS) substitutions, 24 genes (including rbcL and ycf1) display signature consistent with positive selection. When compared with other members of the orchid family, the Ophrys plastome has a complete set of 11 functional ndh plastid genes, with the exception of O. sphegodes that has a truncated ndhF gene. Comparative analysis showed a large co-linearity with other related Orchidinae. However, in contrast to O. iricolor and other Orchidinae, O. sphegodes has a shift of the junction between the Inverted Repeat and Small Single Copy regions associated with the loss of the partial duplicated gene ycf1 and the truncation of the ndhF gene. Data on relative genomic coverage and validation by PCR indicate the presence, with a different ratio, of the two plastome types (i.e. with and without ndhF deletion) in both Ophrys species, with a predominance of the deleted type in O. sphegodes. A search for this deleted plastid region in O. sphegodes nuclear genome shows that the deleted region is inserted in a retrotransposon nuclear sequence. The present study provides useful genomic tools for studying conservation and patterns of relationships of this rapidly radiating orchid genus.


Highly flexible silica/chitosan hybrid scaffolds with oriented pores for tissue regeneration.
Inorganic/organic sol-gel hybrids have nanoscale co-networks of organic and inorganic components that give them the unique potential of tailored mechanical properties and controlled biodegradation in tissue engineering applications. Here, silica/chitosan hybrid scaffolds with oriented structures were fabricated through the sol-gel method with a unidirectional freeze casting process. 3-Glycidoxypropyl trimethoxysilane (GPTMS) was used to obtain covalent inorganic/organic coupling. Process variables were investigated such as cooling rate, GPTMS and inorganic content, which can be used to tailor the mechanical properties and hybrid chemical coupling. Structural characterization and dissolution tests confirmed the covalent cross-linking of the chitosan and the silica network in hybrids. The scaffolds had a directional lamellar structure along the freezing direction and a cellular morphology perpendicular to the freezing direction. Compression testing showed that the scaffolds with 60 wt% organic were flexible and elastomeric perpendicular to the freezing direction whilst behaving in an elastic-brittle fashion parallel to the freezing direction. The compressive strengths are about one order of magnitude higher in the latter direction reaching values of the order of 160 kPa. This behaviour provides potential for clinicians to be able to squeeze the materials to fit tissue defect sites while providing some mechanical support from the other direction.


Automated analysis of single cells using Laser Tweezers Raman Spectroscopy.
In recent years, significant progress has been made into the label-free detection and discrimination of individual cancer cells using Laser Tweezers Raman Spectroscopy (LTRS). However, the majority of examples reported have involved manual trapping of cells, which is time consuming and may lead to different cell lines being analysed in discrete batches. A simple, low-cost microfluidic flow chamber is introduced which allows single cells to be optically trapped and analysed in an automated fashion, greatly reducing the level of operator input required. Two implementations of the flow chamber are discussed here; a basic single-channel device in which the fluid velocity is controlled manually, and a dual-channel device which permits the automated capture and analysis of multiple cell lines with no operator input. Results are presented for the discrimination of live epithelial prostate cells and lymphocytes, together with a consideration of the consequences of traditional 'batch analysis' typically used for LTRS of live cells.


Sensing and capture of toxic and hazardous gases and vapors by metal-organic frameworks.
Toxic and hazardous chemical species are ubiquitous, predominantly emitted by anthropogenic activities, and pose serious risks to human health and the environment. Thus, the sensing and subsequent capture of these chemicals, especially in the gas or vapor phase, are of extreme importance. To this end, metal-organic frameworks have attracted significant interest, as their high porosity and wide tunability make them ideal for both applications. These tailorable framework materials are particularly promising for the specific sensing and capture of targeted chemicals, as they can be designed to fit a diverse range of required conditions. This review will discuss the advantages of metal-organic frameworks in the sensing and capture of harmful gases and vapors, as well as principles and strategies guiding the design of these materials. Recent progress in the luminescent detection of aromatic and aliphatic volatile organic compounds, toxic gases, and chemical warfare agents will be summarized, and the adsorptive removal of fluorocarbons/chlorofluorocarbons, volatile radioactive species, toxic industrial gases and chemical warfare agents will be discussed.


Combined conjugated esterified estrogen plus methyltestosterone supplementation and risk of breast cancer in postmenopausal women.
Testosterone supplementation is being prescribed increasingly to treat symptoms of hormone deficiency in pre- and postmenopausal women; however, studies of the association of testosterone therapy, alone or in combination with estrogen, with risk of breast cancer are limited. The current study assessed the association of combination conjugated esterified estrogen and methyltestosterone (CEE+MT) use and breast cancer risk in postmenopausal women in the Women's Health Initiative (WHI). At Year 3 of follow-up, women in the WHI observational study (N=71,964) provided information on CEE+MT use in the past two years, duration of use, and the brand name of the product. In addition, in each of years 4-8, women were asked whether they had used CEE+MT in the previous year. After 10 years of follow-up, 2832 incident breast cancer cases were identified. Cox proportional hazards models were used to estimate hazard ratios (HR) and 95% confidence intervals (95% CI) for the association of CEE+MT use (irrespective of use of other hormones) and of exclusive CEE+MT use in relation to breast cancer risk. Neither CEE+MT use nor exclusive use of CEE+MT was associated with risk: multivariable-adjusted HR 1.06, 95% CI 0.82-1.36 and HR 1.22, 95% CI 0.78-1.92, respectively. Among women with a natural menopause, the HR for exclusive use was 1.32 (95% CI 0.68-2.55). There was no indication of an association when repeated measures of CEE+MT use were included in a time-dependent covariates analysis. The present study, the largest prospective study to date, did not show a significant association of CEE+MT supplementation and risk of breast cancer.


Specialty bias in treatment recommendations and quality of life among radiation oncologists and urologists for localized prostate cancer.
Given the importance of physician attitudes about different treatments and the quality of life (QOL) in prostate cancer, we performed a national survey of specialists to assess treatment recommendations and perceptions of treatment-related survival and QOL. We mailed a self-administered survey instrument to a random sample of 1366 specialists in the U.S. Respondents were asked for treatment recommendations and survival that varied by PSA levels and Gleason scores and estimate QOL outcomes. Pearson's chi-square and multivariable regression models were used to test for differences in each outcome. Response rates were similar for radiation oncologists (52.6%) and urologists (52.3%; P=0.92). Across all risk strata, urologists were more likely to recommend surgery than were radiation oncologists, for conditions ranging from PSA>20 and Gleason score 8-10 (35.2 vs. 0.2%; P<0.001) to PSA 4-10 and Gleason score 7 (87.5 vs. 20.9%; P<0.001). Radiation oncologists were also more likely to recommend radiation therapy relative to urologists (all P<0.001). From low- to high-risk prostate cancer, radiation oncologists and urologists perceived their treatment as being better for improving survival (all P<0.001). Each specialty also viewed their treatment as having less urinary incontinence (all P<0.001). Radiation oncologists and urologists both prefer the treatment modalities they offer, perceive them to be more effective and to lead to a better QOL. Patients may be receiving biased information, and a truly informed consent process with shared decision-making may be possible only if they are evaluated by both specialties before deciding upon a treatment course.


Emotional distress and compassionate responses in palliative care decision-making consultations.
Seriously ill hospitalized patients and their loved ones are frequently faced with complex treatment decisions laden with expressions of emotional distress during palliative care (PC) consultations. Little is known about these emotional expressions or the compassionate responses providers make and how common these are in PC decision-making conversations. To describe the types and frequency of emotional distress that patients and loved ones express and how providers respond to these emotions during PC decision-making consultations with seriously ill hospitalized patients. We used a quantitative descriptive approach to analyze 71 audio-recorded inpatient PC decision-making consultations for emotional distress and clinicians' responses to those emotions using reliable and established methods. A total of 69% of conversations contained at least one expression of emotional distress. The per-conversation frequency of expressions of emotional distress ranged from 1 to 10. Anxiety/fear were the most frequently encountered emotions (48.4%) followed by sadness (35.5%) and anger/frustration (16.1%). More than half of the emotions related to the patient's feelings (53.6%) and 41.9% were related to the loved ones' own emotions. The majority of emotions were moderate in intensity (65.8%) followed by strong (20.7%) and mild (13.5%). Clinicians responded to a majority of emotions with a compassionate response (75.7%) followed by those with medical content (21.9%) and very few were ignored (1.3%). Expressions of emotional distress are common during PC consultations and are usually met with compassionate responses by the clinician.


Modeling Potential-Dependent Electrochemical Activation Barriers: Revisiting the Alkaline Hydrogen Evolution Reaction.
Accurate theoretical simulation of electrochemical activation barriers is key to understanding electrocatalysis and guides the design of more efficient catalysts. Providing a detailed picture of proton transfer processes encounters several challenges: the constant potential requirement during charge transfer, the different time scales involved in the processes, and the thermal fluctuation of the solvent. Hence, it is prohibitively expensive computationally to apply density functional theory (DFT) calculations in modeling the potential-dependent activation barrier at the electrode-solvent interface, and the results are dubious. To address these challenges, we have developed an analytical approach based on charge conservation and decoupled potential energy surfaces to compute charge transfer barriers. The method makes it possible to simulate an electrochemical process at different potentials and explicitly include thermal fluctuations of the solvent at the electrode-solvent interface. We use the Pt-catalyzed alkaline hydrogen evolution reaction (HER) as our benchmark reaction, and we model the microkinetics of HER with consideration of the spatial fluctuations between the metal surface and the first solvent layer at room temperature. The distribution of water-metal distances has a large effect on the barriers of the charge transfer processes, and an accurate account of the statistical fluctuation in the reaction network leads to a several orders of magnitude increase in HER current as compared to transfer from a static solvent. The trends of the different reaction mechanisms in HER were successfully simulated with our model, and the theoretical I-V curves obtained are in good qualitative agreement with experimental results.


Hybrid Nitric Oxide Donor and its Carrier for the Treatment of Peripheral Arterial Diseases.
Nitric oxide (NO) has been known to promote physiological angiogenesis to treat peripheral arterial diseases (PAD) by increasing the vascular endothelial growth factor (VEGF) level in endothelial cells (ECs) and preventing platelet adherence and leukocyte chemotaxis. However, the ongoing ischemic event during peripheral ischemia produces superoxide and diminishes the NO bioavailability by forming toxic peroxynitrite anion. Here we disclose an efficacious hybrid molecule 4-(5-Amino-1,2,3-oxadiazol-3-yl)-2,2,6,6-tetramethyl-1-piperidinol (SA-2) containing both antioxidant and NO donor functionalities that provide a therapeutic level of NO necessary to promote angiogenesis and to protect ECs against hydrogen peroxide-induced oxidative stress. Compound SA-2 scavenged reactive oxygen species, inhibited proliferation and migration of smooth muscle cells (SMCs) and promoted the tube formation from ECs. Copolymer poly(lactic-co-glycolic acid) (PLGA) nanoparticles loaded with SA-2 provided a sustained release of NO over days, improved aqueous stability in serum, protected ECs against oxidative stress, and enhanced angiogenesis under stress conditions as compared to that of the control in the in vitro matrigel tube formation assay. These results indicated the potential use of SA-2 nanoparticles as an alternative therapy to treat PAD.


Computer-assisted instruction as a learning resource for applied anatomy and kinesiology in the occupational therapy curriculum.
The purpose of these studies was to examine the learning outcomes of a computer-assisted instruction (CAI) tutorial in applied anatomy and kinesiology for occupational therapy students and to determine its applicability for use in two university settings. Two separate pilot studies were conducted at two universities. In each study, the learning outcomes of an experimental group of occupational therapy students using a CAI program and a control group using books to study the same material were compared. Learning outcomes were assessed with post-test achievement test scores on an applied anatomy and kinesiology test and responses to an attitude questionnaire with Likert-scale items and open-ended questions. There was no significant difference in the means on achievement test scores for the experimental and control groups in the first pilot study. In the second study, the CAI group scored significantly higher on the achievement test than the control group. In both pilot studies, subjects displayed significantly more positive attitudes toward the CAI program as a learning tool than they did toward traditional self-study with books. A CAI program in applied anatomy and kinesiology can be an effective supplemental resource for occupational therapy students and can offer a learning experience that students value and perceive as helpful. Establishment of clear learning objectives, use of a theoretical base to design instruction, and development and testing in different educational settings can help improve the quality of CAI programs and ensure their relevance to other curricula.


Passive Immunotherapy: Assessment of Convalescent Serum Against Ebola Virus Makona Infection in Nonhuman Primates.
Convalescent serum and blood were used to treat patients during outbreaks of Zaire ebolavirus (ZEBOV) infection in 1976 and 1995, with inconclusive results. During the recent 2013-2016 West African epidemic, serum/plasma from survivors of ZEBOV infection was used to treat patients in the affected countries and several repatriated patients. The effectiveness of this strategy remains unknown. Nine rhesus monkeys were experimentally infected with ZEBOV-Makona. Beginning on day 3 after exposure (at the onset of viremia), 4 animals were treated with homologous ZEBOV-Makona convalescent macaque sera, 3 animals were treated in parallel with heterologous Sudan ebolavirus (SEBOV) convalescent macaque sera, and 2 animals served as positive controls and were not treated. Surviving animals received additional treatments on days 6 and 9. Both untreated control animals died on postinfection day 9. All 4 ZEBOV-Makona-infected macaques treated with homologous ZEBOV-Makona convalescent sera died on days 8-9. One macaque treated with heterologous SEBOV convalescent sera survived, while the other animals treated with the heterologous SEBOV sera died on days 7 and 9. The findings suggest that convalescent sera alone is not sufficient for providing 100% protection against lethal ZEBOV infection when administered at the onset of viremia.


Abnormal breath tests to lactose, fructose and sorbitol in irritable bowel syndrome may be explained by small intestinal bacterial overgrowth.
Small intestinal bacterial overgrowth and sugar malabsorption (lactose, fructose, sorbitol) may play a role in irritable bowel syndrome. The lactulose breath test is a reliable and non-invasive test for the diagnosis of small intestinal bacterial overgrowth. The lactose, fructose and sorbitol hydrogen breath tests are widely used to detect specific sugar malabsorption. To assess the extent to which small intestinal bacterial overgrowth may influence the results of hydrogen sugar breath tests in irritable bowel syndrome patients. We enrolled 98 consecutive irritable bowel syndrome patients. All subjects underwent hydrogen lactulose, lactose, fructose and sorbitol hydrogen breath tests. Small intestinal bacterial overgrowth patients were treated with 1-week course of antibiotics. All tests were repeated 1 month after the end of therapy. A positive lactulose breath test was found in 64 of 98 (65%) subjects; these small intestinal bacterial overgrowth patients showed a significantly higher prevalence of positivity to the lactose breath test (P < 0.05), fructose breath test (P < 0.01) and sorbitol breath test (P < 0.01) when compared with the small intestinal bacterial overgrowth-negatives. Small intestinal bacterial overgrowth eradication, as confirmed by negative lactulose breath test, caused a significant reduction in lactose, fructose and sorbitol breath tests positivity (17% vs. 100%, 3% vs. 62%, and 10% vs. 71% respectively: P < 0.0001). In irritable bowel syndrome patients with small intestinal bacterial overgrowth, sugar breath tests may be falsely abnormal. Eradication of small intestinal bacterial overgrowth normalizes sugar breath tests in the majority of patients. Testing for small intestinal bacterial overgrowth should be performed before other sugar breath tests tests to avoid sugar malabsorption misdiagnosis.


Increase in interstitial interleukin-6 of human skeletal muscle with repetitive low-force exercise.
Interleukin (IL)-6, which is released from muscle tissue during intense exercise, possesses important metabolic and probably anti-inflammatory properties. To evaluate the IL-6 response to low-intensity exercise, we conducted two studies: 1) a control study with insertion of microdialysis catheters in muscle and determination of interstitial muscle IL-6 response over 2 h of rest and 2) an exercise study to investigate the IL-6 response to 20 min of repetitive low-force exercise. In both studies, a microdialysis catheter (cutoff: 3,000 kDa) was inserted into the upper trapezius muscle of six male subjects, and the catheters were perfused with Ringer-acetate at 5 microl/min. Venous plasma samples were taken in the exercise study. The insertion of microdialysis catheters into muscle resulted in an increase in IL-6 from 8 +/- 0 to 359 +/- 171 and 484 +/- 202 pg/ml after 65 and 110 min, respectively (P < 0.001). Similarly, in the exercise study, IL-6 increased to 289 +/- 128 pg/ml after a 55-min rest (P < 0.001). During the subsequent repetitive low-force exercise, muscle IL-6 further increased to 1,246 +/- 461 pg/ml and reached 2,132 +/- 477 pg/ml after a 30-min recovery (all P < 0.001). In contrast to this, plasma IL-6 did not significantly change in response to exercise. We conclude that upper extremity, low-intensity exercise results in a substantial increase in IL-6 in the interstitium of the stabilizing trapezius muscle, whereas no change is seen for plasma IL-6.


Paradoxical relationship between frequency selectivity and threshold sensitivity during auditory-nerve fiber development.
The acquisition of adult-like frequency selectivity is generally assumed to be the tightly coupled to improvements in threshold sensitivity during cochlear development. In this study, frequency versus threshold (tuning) curves obtained from 1108 auditory-nerve fibers were used to investigate the relationship between tuning and threshold at characteristic frequency (CF) during postnatal development in kittens. At the earliest ages included in this study, sharpness was within the adult range, but thresholds were significantly higher than adult values. Tuning and thresholds improved along different exponential time courses that varied with CF. For units with CFs below 1 kHz, tuning curve slopes below CF matured earliest, followed by CF threshold, and then by slopes above CF. In contrast, for CFs above 1 kHz, the high-frequency slopes matured first, followed by threshold and then by slope below CF. One interpretation of these results is that tuning and thresholds are not tightly coupled in immature animals. Paradoxically, however, high-frequency slopes were correlated with threshold for individual units at all ages, suggesting that the relationship between tuning and threshold is maintained during development. This contradiction can be resolved by a developmental model that features a functional separation between cochlear nonlinearities and mechanical/electrical conversion.


Results of a phase 1 study of AME-133v (LY2469298), an Fc-engineered humanized monoclonal anti-CD20 antibody, in FcγRIIIa-genotyped patients with previously treated follicular lymphoma.
AME-133v is a humanized monoclonal antibody engineered to have increased affinity to CD20 and mediate antibody-dependent cell-mediated cytotoxicity (ADCC) better than rituximab. Safety, pharmacokinetics, and efficacy were assessed in a phase 1/2 trial in patients with previously treated follicular lymphoma (FL). AME-133v was characterized in vitro by ADCC and cell binding assays. A phase 1 study was conducted in which 23 previously treated patients with FL were assigned sequentially to one of five dose-escalation cohorts of AME-133v at 2, 7.5, 30, 100, or 375 mg/m(2) weekly × 4 doses. AME-133v showed a 13- to 20-fold greater binding affinity for CD20 and was 5- to 7-fold more potent than rituximab in ADCC assays. Cell binding assays showed AME-133v and rituximab competed for an overlapping epitope on the CD20 antigen, and AME-133v inhibited binding of biotinylated rituximab to CD20 in a concentration-dependent manner. AME-133v was well tolerated by patients and common related adverse events included chills and fatigue. One patient experienced a dose-limiting toxicity of neutropenia. AME-133v showed nonlinear pharmocokinetics with properties similar to rituximab. Selective reduction of B cells during and after AME-133v treatment was shown by flow cytometry of peripheral blood. A partial or complete response was observed in 5 of 23 (22%) patients and the median progression-free survival was 25.4 weeks. AME-133v was safe and well tolerated at the doses tested. AME-133v showed encouraging results as an anti-CD20 therapy in heavily pretreated FL patients with the less favorable FcγRIIIa F-carrier genotype.


Carrier detection of Duchenne muscular dystrophy through analysis of DNA from deciduous teeth of a dead affected child.
The sister of a child affected by Duchenne muscular dystrophy (DMD) was referred for genetic counselling to assess the risk of her being a carrier. Her brother had died 15 years previously at the age of 8. There were no other affected males in the family. There were no methods for DNA investigation at the time of the child's death and the family had never been studied for linkage with polymorphic probes on the chromosomal region Xp21. The only tissue from which an assessment of the risk could be made by DNA linkage analysis was two of the child's deciduous teeth that the parents had kept. DNA was extracted using a protocol described for the recovery of ancient DNA from museum specimens and archaeological finds. Multiplex amplification did not reveal deletions in 19 exons spanning the hot-spot regions for deletions within the dystrophin gene in Xp21. Linkage analysis using three highly polymorphic microsatellites demonstrated that the sister had not received the X chromosome borne by her brother. These results show that DNA extracted from teeth is a reliable source for molecular diagnosis.


Increased leukotriene B4 release from ileal pouch mucosa in ulcerative colitis compared with familial adenomatous polyposis.
Pouchitis may complicate the construction of an ileal pouch after colectomy for ulcerative colitis (UC) but not familial adenomatous polyposis (FAP). To examine whether differences in eicosanoid metabolism might explain why pouchitis is largely confined to UC patients, this study compared arachidonic acid stimulated release of immunoreactive leukotriene B4 (LTB4) and prostaglandin E2 (PGE2) from macroscopically uninflamed pouch mucosal biopsy specimens incubated in vitro from patients with UC and FAP. The study also compared eicosanoid release from inflamed and uninflamed pouches in patients with UC. In uninflamed pouches, median LTB4 release was nearly twice as high in UC as in FAP (p = 0.001), but there was no significant difference in PGE2 production. In UC, stimulated eicosanoid release from uninflamed functioning pouch mucosa was not significantly different from that from either ileostomy or defunctioned pouch mucosa. LTB4 and PGE2 release were significantly greater from inflamed than uninflamed pouch mucosa in UC (p = 0.001 and 0.01, respectively). Leukotriene synthesis inhibition or receptor antagonism, or both merit therapeutic evaluation in pouchitis. Increased release of LTB4 from endoscopically normal pouch mucosa suggests increased 5-lipoxygenase activity in patients with UC and could contribute to their predisposition to pouchitis.


Depth profiles of hydrogen bound water molecule types and their relation to lipid and protein interaction in the human stratum corneum in vivo.
Confocal Raman microscopy has been used to measure depth-dependent profiles of human SC in vivo in the high wavenumber (HWN) region. In order to keep the linearity of HWN region boundaries and to not remove an informative signal from Raman spectra, a new baseline subtraction procedure has been introduced. After baseline subtraction, the HWN spectrum was deconvoluted using 10 Gaussian functions with individual chemical meanings. The results show that the hydrogen bound water molecule types contributed differently to the water diffusion process in the SC. The most concentrated double donor-double acceptor (DDAA) and single donor-single acceptor (DA) water molecule types in the SC represent more than 90% of the SC's water and mostly contribute to the water flux in the skin. Single donor-double acceptor (DAA) and weakly-bound water molecule types represent less than 10% of the SC's water content. The most tightly hydrogen bound water molecule type, DAA, reaches its maximum concentration near the skin surface and does not take part in the water diffusion process via the SC. The results show that the hydrogen bonding state of water (DA/DDAA water molecule type ratio) reaches its maximum at the depth of approx. 30% of the SC thickness, which correlates well with the maximum lateral packing order of intercellular lipids (ICL) and the natural moisturizing factor (NMF), and does not coincide with the folding/unfolding state of keratin. The NMF's contribution to the bonding of water in the SC is supposed to dominate over that of ICL.


Are there predictors that can determine neoadjuvant treatment responses in rectal cancer?
This study aimed to determine a predictive bioindicator that would detect the treatment response of patients diagnosed with rectal cancer and treated with neoadjuvant chemoradiotherapy (nCRT). The data collected from 37 patients receiving nCRT were retrospectively evaluated. The p53 score and gene instability in MLH1 and MSH2, which are among the DNA mismatch repair (MMR) genes, were evaluated using immunohistochemical methods. The neutrophils-leukocytes ratio (NLR), carcinoembryonic antigen (CEA), and carbohydrate antigen (CA) 19-9 values were obtained as hematological parameters from computer records. The pathologic analysis of the therapy response after nCRT was classified according to the modified grading system by Ryan et al. Results: The changes in the NLR, CEA, and CA19-9 values before and after treatment were statistically significant (p<0.001 and p=0.005). A near significant effect of the decrease of the CEA value in the 5th week after treatment was detected on the pathological response score (p=0.075). The p53 mutation score in those patients with any residue was higher than the total response. Overall, 89.2% of the patients exhibited MMR positivity (stability), and 10.8% of the cases with MRM negativity (instability) had a macroscopic residue. Cases with pathological total response were MRM positive. Consequently, in most of the patients treated with nCRT, the treatment caused tumor and nodal remission. In the prediction of this therapy response, hematological and genetic parameters, such as NLR, P53, MLH1, and MSH2, play a predictive role.


Persistence of butterfly populations in fragmented habitats along urban density gradients: motility helps.
In a simulation study of genotypes conducted over 100 generations for more than 1600 butterfly's individuals, we evaluate how the increase of anthropogenic fragmentation and reduction of habitat size along urbanisation gradients (from 7 to 59% of impervious land cover) influences genetic diversity and population persistence in butterfly species. We show that in areas characterised by a high urbanisation rate (>56% impervious land cover), a large decrease of both genetic diversity (loss of 60-80% of initial observed heterozygosity) and population size (loss of 70-90% of individuals) is observed over time. This is confirmed by empirical data available for the mobile butterfly species Pieris rapae in a subpart of the study area. Comparing simulated data for P. rapae with its normal dispersal ability and with a reduced dispersal ability, we also show that a higher dispersal ability can be an advantage to survive in an urban or highly fragmented environment. The results obtained here suggest that it is of high importance to account for population persistence, and confirm that it is crucial to maintain habitat size and connectivity in the context of land-use planning.


SMARCAL1 maintains telomere integrity during DNA replication.
The SMARCAL1 (SWI/SNF related, matrix-associated, actin-dependent, regulator of chromatin, subfamily A-like 1) DNA translocase is one of several related enzymes, including ZRANB3 (zinc finger, RAN-binding domain containing 3) and HLTF (helicase-like transcription factor), that are recruited to stalled replication forks to promote repair and restart replication. These enzymes can perform similar biochemical reactions such as fork reversal; however, genetic studies indicate they must have unique cellular activities. Here, we present data showing that SMARCAL1 has an important function at telomeres, which present an endogenous source of replication stress. SMARCAL1-deficient cells accumulate telomere-associated DNA damage and have greatly elevated levels of extrachromosomal telomere DNA (C-circles). Although these telomere phenotypes are often found in tumor cells using the alternative lengthening of telomeres (ALT) pathway for telomere elongation, SMARCAL1 deficiency does not yield other ALT phenotypes such as elevated telomere recombination. The activity of SMARCAL1 at telomeres can be separated from its genome-maintenance activity in bulk chromosomal replication because it does not require interaction with replication protein A. Finally, this telomere-maintenance function is not shared by ZRANB3 or HLTF. Our results provide the first identification, to our knowledge, of an endogenous source of replication stress that requires SMARCAL1 for resolution and define differences between members of this class of replication fork-repair enzymes.


salvador Promotes both cell cycle exit and apoptosis in Drosophila and is mutated in human cancer cell lines.
The number of cells in an organism is determined by regulating both cell proliferation and cell death. Relatively few mechanisms have been identified that can modulate both of these processes. In a screen for Drosophila mutations that result in tissue overgrowth, we identified salvador (sav), a gene that promotes both cell cycle exit and cell death. Elevated Cyclin E and DIAP1 levels are found in mutant cells, resulting in delayed cell cycle exit and impaired apoptosis. Salvador contains two WW domains and binds to the Warts (or LATS) protein kinase. The human ortholog of salvador (hWW45) is mutated in three cancer cell lines. Thus, salvador restricts cell numbers in vivo by functioning as a dual regulator of cell proliferation and apoptosis.


Current management of hilar cholangiocarcinoma.
The goal in the treatment of hilar cholangiocarcinoma is the relief of biliary obstruction with the quality survival. The choice of therapy for an individual patient requires complete preoperative staging to determine the resectability. This is best accomplished in a multidisciplinary setting with radiologists, gastroenterologists and surgeons participating in the diagnostic and therapeutic strategies. Operative removal of the tumor can be performed with low mortality and, when possible, provides the longest palliation and offers potential for cure. For tumors found at operation to be unresectable, biliary enteric bypass provides a durable method of palliation. The relative merits of the newer methods of radiologic stenting are encouraging, but further study will require prospective trials in comparison to operative biliary enteric bypass.


Using Predictive Analytics for Early Identification of Short-Term Disability Claimants Who Exhaust Their Benefits.
Purpose Early interventions can help short-term disability insurance (STDI) claimants return to work following onset of an off-the-job medical condition. Accurately targeting such interventions involves identifying claimants who would, without intervention, exhaust STDI benefits and transition to longer-term support. We identify factors that predict STDI exhaustion and transfer to long-term disability insurance (LTDI). We also explore whether waiting for some claims to resolve without intervention improves targeting efficiency. Methods We use a large database of STDI claims from private employer-sponsored disability insurance programs in the United States to predict which claims will exhaust STDI or transition to LTDI. We use a split sample approach, conducting logistic regressions on half of our data and generating predictions for the other half. We assess predictive accuracy using ROC curve analysis, repeating on successive subsamples, omitting claims that resolve within 2, 4, and 6 weeks. Results Age, primary diagnosis, and employer industry were associated with the two outcomes. Rapid attrition of short-duration claims from the sample means that waiting can substantially increase the efficiency of targeting efforts. Overall accuracy of classification increases from 63.2% at week 0 to 82.9% at week 6 for exhausting STDI benefits, and from 63.7 to 83.0% for LTDI transfer. Conclusions Waiting even a few weeks can substantially increase the accuracy of early intervention targeting by allowing claims that will resolve without further intervention to do so. Predictive modeling further narrows the target population based on claim characteristics, reducing intervention costs. Before adopting a waiting strategy, however, it is important to consider potential trade-offs involved in delaying the start of any intervention.


Comparison of two simulation systems to support robotic-assisted surgical training: a pilot study (Swine model).
To compare the efficacy of simulation-based training between the Mimic dV- Trainer and traditional dry lab da Vinci robot training. A prospective randomized study analyzing the performance of 20 robotics-naive participants. Participants were enrolled in an online da Vinci Intuitive Surgical didactic training module, followed by training in use of the da Vinci standard surgical robot. Spatial ability tests were performed as well. Participants were randomly assigned to 1 of 2 training conditions: performance of 3 Fundamentals of Laparoscopic Surgery dry lab tasks using the da Vinci or performance of 4 dV-Trainer tasks. Participants in both groups performed all tasks to empirically establish proficiency criterion. Participants then performed the transfer task, a cystotomy closure using the daVinci robot on a live animal (swine) model. The performance of robotic tasks was blindly assessed by a panel of experienced surgeons using objective tracking data and using the validated Global Evaluative Assessment of Robotic Surgery (GEARS), a structured assessment tool. No statistically significant difference in surgeon performance was found between the 2 training conditions, dV-Trainer and da Vinci robot. Analysis of a 95% confidence interval for the difference in means (-0.803 to 0.543) indicated that the 2 methods are unlikely to differ to an extent that would be clinically meaningful. Based on the results of this study, a curriculum on the dV- Trainer was shown to be comparable to traditional da Vinci robot training. Therefore, we have identified that training on a virtual reality system may be an alternative to live animal training for future robotic surgeons.


Static and dynamic selection thresholds governing the accumulation of information in genetic algorithms using ranked populations.
Mutation applied indiscriminately across a population has, on average, a detrimental effect on the accumulation of solution alleles within the population and is usually beneficial only when targeted at individuals with few solution alleles. Many common selection techniques can delete individuals with more solution alleles than are easily recovered by mutation. The paper identifies static and dynamic selection thresholds governing accumulation of information in a genetic algorithm (GA). When individuals are ranked by fitness, there exists a dynamic threshold defined by the solution density of surviving individuals and a lower static threshold defined by the solution density of the information source used for mutation. Replacing individuals ranked below the static threshold with randomly generated individuals avoids the need for mutation while maintaining diversity in the population with a consequent improvement in population fitness. By replacing individuals ranked between the thresholds with randomly selected individuals from above the dynamic threshold, population fitness improves dramatically. We model the dynamic behavior of GAs using these thresholds and demonstrate their effectiveness by simulation and benchmark problems.


Cytometry by time-of-flight shows combinatorial cytokine expression and virus-specific cell niches within a continuum of CD8+ T cell phenotypes.
Cytotoxic CD8(+) T lymphocytes directly kill infected or aberrant cells and secrete proinflammatory cytokines. By using metal-labeled probes and mass spectrometric analysis (cytometry by time-of-flight, or CyTOF) of human CD8(+) T cells, we analyzed the expression of many more proteins than previously possible with fluorescent labels, including surface markers, cytokines, and antigen specificity with modified peptide-MHC tetramers. With 3-dimensional principal component analysis (3D-PCA) to display phenotypic diversity, we observed a relatively uniform pattern of variation in all subjects tested, highlighting the interrelatedness of previously described subsets and the continuous nature of CD8(+) T cell differentiation. These data also showed much greater complexity in the CD8(+) T cell compartment than previously appreciated, including a nearly combinatorial pattern of cytokine expression, with distinct niches occupied by virus-specific cells. This large degree of functional diversity even between cells with the same specificity gives CD8(+) T cells a remarkable degree of flexibility in responding to pathogens.


Effect of prosocial public health messages for population behaviour change in relation to respiratory infections: a systematic review protocol.
The COVID-19 pandemic represents a major societal challenge that requires large-scale behaviour change, widespread collective action and cooperation to reduce viral transmission. Existing literature indicates that several messaging approaches may be effective, including emphasising the benefits to the recipient, aligning with the recipient's moral values and focusing on protecting others. Current research suggests that prosocial public health messages that highlight behaviours linked to societal benefits (eg, protecting 'each other'), rather than focusing on behaviours that protect oneself (eg, protecting 'yourself'), may be a more effective method for communicating strategies related to infectious disease. To investigate this we will conduct a systematic review that will identify what messages and behaviour change techniques have the potential to optimise the effect on population behaviour in relation to reducing transmission of respiratory infections. A systematic literature search of published and unpublished studies (including grey literature) in electronic databases will be conducted to identify those that meet our inclusion criteria. The search will be run in four electronic databases: MEDLINE, EMBASE, PsycINFO and Scopus. We will also conduct supplementary searches in databases of 'grey' literature such as PsycEXTRA, Social Science Research Network and OSF PREPRINTS, and use the Google Scholar search engine. A systematic approach to searching, screening, reviewing and data extraction will be applied based on Preferred Reporting Items for Systematic Reviews and Meta-Analyses. Titles, abstracts and full texts for eligibility will be examined independently by researchers. The quality of the included studies will be assessed using the Cochrane Risk of Bias Tool and the Risk of Bias in Non-randomized Studies-of Interventions tool. Disagreements will be resolved by a consensus procedure. This protocol has been registered with PROSPERO. No ethical approval is required, as there will be no collection of primary data. The synthesised findings will be disseminated through peer-reviewed publication. CRD42020198874.


Conjugated Polymer-Small Molecule Alloy Leads to High Efficient Ternary Organic Solar Cells.
Ternary organic solar cells are promising candidates for bulk heterojunction solar cells; however, improving the power conversion efficiency (PCE) is quite challenging because the ternary system is complicated on phase separation behavior. In this study, a ternary organic solar cell (OSC) with two donors, including one polymer (PTB7-Th), one small molecule (p-DTS(FBTTH2)2), and one acceptor (PC71BM), is fabricated. We propose the two donors in the ternary blend forms an alloy. A notable averaged PCE of 10.5% for ternary OSC is obtained due to the improvement of the fill factor (FF) and the short-circuit current density (J(sc)), and the open-circuit voltage (V(oc)) does not pin to the smaller V(oc) of the corresponding binary blends. A highly ordered face-on orientation of polymer molecules is obtained due to the formation of an alloy structure, which facilitates the enhancement of charge separation and transport and the reduction of charge recombination. This work indicates that a high crystallinity and the face-on orientation of polymers could be obtained by forming alloy with two miscible donors, thus paving a way to largely enhance the PCE of OSCs by using the ternary blend strategy.


Structure-activity relationship of a broad-spectrum insect odorant receptor agonist.
Agonism of insect odorant receptor (OR) cation channels may represent a new strategy for the manipulation of destructive insect olfactory-driven behaviors. We have explored the chemical space around VUAA1, the first in class agonist of the obligate OR co-receptor ion channel (Orco), and describe novel compound analogues with increased potency across insect taxa. Functional analyses reveal several of these VUAA1 structural analogues display significantly greater potency as compared to the activity of the previously described active compounds in mobility-based behavioral assays on mosquito larvae.


Comparative evaluation of conventional and microwave hydrothermal carbonization of human biowaste for value recovery.
This paper compares conventional and microwave hydrothermal carbonization (HTC) of human biowaste (HBW) at 160 °C, 180 °C and 200 °C as a potential technology to recover valuable carbonaceous solid fuel char and organic-rich liquor. Also discussed are the influence of HTC heating methods and temperature on HBW processing conversion into solid fuel char, i.e. yield and post-HTC management, dewaterability rates, particle size distribution and the carbon and energy properties of solid fuel char. While HTC temperatures influenced all parameters investigated, especially yield and properties of end products recovered, heating source effects were noticeable on dewatering rates, char particle sizes and HBW processing/end product recovery rate and, by extension, energy consumed. The microwave process was found to be more efficient for dewatering processed HBW and for char recovery, consuming half the energy used by the conventional HTC method despite the similarity in yields, carbon and energy properties of the recovered char. However, both processes reliably overcame the heterogeneity of HBW, converting them into non-foul end products, which were easily dewatered at <3 seconds/g total solids (TS) (c.f. 50.3 seconds/g TS for a raw sample) to recover energy-densified chars of ≈17 MJ/kg calorific value and up to 1.4 g/l of ammonia concentration in recovered liquor.


All-cause mortality in HIV-positive adults starting combination antiretroviral therapy: correcting for loss to follow-up.
To estimate mortality in HIV-positive patients starting combination antiretroviral therapy (ART) and to discuss different approaches to calculating correction factors to account for loss to follow-up. A total of 222 096 adult HIV-positive patients who started ART 2009-2014 in clinics participating in the International epidemiology Databases to Evaluate AIDS collaboration in 43 countries in sub-Saharan Africa, Asia Pacific, Latin America, and North America were included. To allow for underascertainment of deaths due to loss to follow-up, two correction factors (one for the period 0-6 months on ART and one for later periods) or 168 correction factors (combinations of two sexes, three time periods after ART initiation, four age groups, and seven CD4 groups) based on tracing patients lost in Kenya and data linkages in South Africa were applied. Corrected mortality rates were compared with a worst case scenario assuming all patients lost to follow-up had died. Loss to follow-up differed between regions; rates were lowest in central Africa and highest in east Africa. Compared with using two correction factors (1.64 for the initial ART period and 2.19 for later), applying 168 correction factors (range 1.03-4.75) more often resulted in implausible mortality rates that exceeded the worst case scenario. Corrected mortality rates varied widely, ranging from 0.2 per 100 person-years to 54 per 100 person-years depending on region and covariates. Implausible rates were less common with the simpler approach based on two correction factors. The corrected mortality rates will be useful to international agencies, national programmes, and modellers.


Social and environmental predictors of plasma HIV RNA rebound among injection drug users treated with antiretroviral therapy.
Evidence is needed to improve HIV treatment outcomes for individuals who use injection drugs (IDU). Although studies have suggested higher rates of plasma viral load (PVL) rebound among IDU on antiretroviral therapy (ART), risk factors for rebound have not been thoroughly investigated. We used data from a long-running community-recruited prospective cohort of IDU in Vancouver, Canada, linked to comprehensive ART and clinical monitoring records. Using proportional hazards methods, we modeled the time to confirmed PVL rebound above 1000 copies per milliliter among IDU on ART with sustained viral suppression, defined as 2 consecutive undetectable PVL measures. Between 1996 and 2009, 277 individuals had sustained viral suppression. Over a median follow-up of 32 months, 125 participants (45.1%) experienced at least 1 episode of virologic failure for an incidence of 12.6 [95% confidence interval (CI): 10.5 to 15.0] per 100 person-years. In a multivariate model, PVL rebound was independently associated with sex-trade involvement [adjusted hazard ratio (AHR) = 1.40, 95% CI: 1.08 to 1.82) and recent incarceration (AHR = 1.83, 95% CI: 1.33 to 2.52). Methadone maintenance therapy (AHR = 0.79, 95% CI: 0.66 to 0.94) was protective. No measure of illicit drug use was predictive. In this setting of free ART, several social and environmental factors predicted higher risks of viral rebound among IDU, including sex-trade involvement and incarceration. These findings should help inform efforts to identify individuals at risk of viral rebound and targeted interventions to treat and retain individuals in effective ART.


Mechanisms of divergence and convergence of the human immunoglobulin alpha 1 and alpha 2 constant region gene sequences.
Nucleotide sequences of the human alpha 1 and two allelic alpha 2 immunoglobulin heavy chain constant region genes are presented. The genes contain three exons, each encoding a single constant region protein domain. The protein hinge region is encoded at the 5' end of the second exon, and the rapid evolutionary changes in length of the hinge correspond to duplications or deletions within the hinge-coding region, probably facilitated by repeats in the DNA sequence. Alignment of the alpha 1 and alpha 2 gene sequences reveals an unusual coupled deletion-duplication in the 5'-flanking region, which can be explained in terms of a slipped-strand mispairing model. Comparison of nucleotide sequences of the alpha 1 gene and two alleles of the alpha 2 gene indicates a localized transfer of genetic information from the 3' end of the alpha 1 gene to one of the alpha 2 alleles, probably by a gene conversion. At one end of the region within which conversion apparently occurred, there is a 40 bp sequence of the type that can form Z-DNA.


Targeted cytotoxic somatostatin analogue AN-238 inhibits somatostatin receptor-positive experimental colon cancers independently of their p53 status.
The resistance of advanced colorectal cancers to therapy is often related to mutations in the p53 tumor suppressor gene. Because somatostatin (SRIF) receptors (ssts) are present in colorectal carcinomas, the treatment with targeted cytotoxic SRIF analogue AN-238, consisting of 2-pyrrolinodoxorubicin (AN-201) linked to octapeptide SRIF carrier RC-121, may overcome this resistance by producing a higher concentration of the cytotoxic agent in the tumors. Four colon cancer cell lines, HCT-116 and LoVo expressing wild-type p53, and HCT-15 and HT-29 with mutated p53, were investigated. HCT-116, HCT-15, and HT-29, but not LoVo possess functional ssts. We analyzed changes in p53, p21, and proliferating cell nuclear antigen (PCNA) concentrations in these cells in vitro by immunoblotting after exposure to AN-238, its radical AN-201, or doxorubicin (DOX). Equitoxic doses of AN-238, AN-201, or DOX affected p53, p21, and PCNA differently. Analysis of the p21:p53 ratios revealed that DOX increased p53 levels, but most of p53 was mutated and inactive, whereas AN-238 produced smaller changes in p53 concentrations but enhanced its activity. In HCT-15 cells, PCNA:p21 ratios, which are indicators of proliferation and repair processes, remained unchanged after exposure to AN-238 but were increased by DOX. In vivo studies in nude mice demonstrated that AN-238, AN-201, and DOX were equally effective on HCT-116 tumors that express wild-type p53. However, AN-238 also inhibited the growth of HCT-15 and HT-29 cancers that express mutant p53, whereas AN-201 and DOX showed no effect. None of the compounds could suppress the proliferation of LoVo tumors that lack functional ssts. In conclusion, cytotoxic SRIF analogue AN-238 inhibits the growth of experimental colon cancers that express ssts, regardless of their p53 status.


Hydrogen sulfide and nitric oxide are mutually dependent in the regulation of angiogenesis and endothelium-dependent vasorelaxation.
Hydrogen sulfide (H(2)S) is a unique gasotransmitter, with regulatory roles in the cardiovascular, nervous, and immune systems. Some of the vascular actions of H(2)S (stimulation of angiogenesis, relaxation of vascular smooth muscle) resemble those of nitric oxide (NO). Although it was generally assumed that H(2)S and NO exert their effects via separate pathways, the results of the current study show that H(2)S and NO are mutually required to elicit angiogenesis and vasodilatation. Exposure of endothelial cells to H(2)S increases intracellular cyclic guanosine 5'-monophosphate (cGMP) in a NO-dependent manner, and activated protein kinase G (PKG) and its downstream effector, the vasodilator-stimulated phosphoprotein (VASP). Inhibition of endothelial isoform of NO synthase (eNOS) or PKG-I abolishes the H(2)S-stimulated angiogenic response, and attenuated H(2)S-stimulated vasorelaxation, demonstrating the requirement of NO in vascular H(2)S signaling. Conversely, silencing of the H(2)S-producing enzyme cystathionine-γ-lyase abolishes NO-stimulated cGMP accumulation and angiogenesis and attenuates the acetylcholine-induced vasorelaxation, indicating a partial requirement of H(2)S in the vascular activity of NO. The actions of H(2)S and NO converge at cGMP; though H(2)S does not directly activate soluble guanylyl cyclase, it maintains a tonic inhibitory effect on PDE5, thereby delaying the degradation of cGMP. H(2)S also activates PI3K/Akt, and increases eNOS phosphorylation at its activating site S1177. The cooperative action of the two gasotransmitters on increasing and maintaining intracellular cGMP is essential for PKG activation and angiogenesis and vasorelaxation. H(2)S-induced wound healing and microvessel growth in matrigel plugs is suppressed by pharmacological inhibition or genetic ablation of eNOS. Thus, NO and H(2)S are mutually required for the physiological control of vascular function.


MTAP deletion confers enhanced dependency on the PRMT5 arginine methyltransferase in cancer cells.
The discovery of cancer dependencies has the potential to inform therapeutic strategies and to identify putative drug targets. Integrating data from comprehensive genomic profiling of cancer cell lines and from functional characterization of cancer cell dependencies, we discovered that loss of the enzyme methylthioadenosine phosphorylase (MTAP) confers a selective dependence on protein arginine methyltransferase 5 (PRMT5) and its binding partner WDR77. MTAP is frequently lost due to its proximity to the commonly deleted tumor suppressor gene, CDKN2A. We observed increased intracellular concentrations of methylthioadenosine (MTA, the metabolite cleaved by MTAP) in cells harboring MTAP deletions. Furthermore, MTA specifically inhibited PRMT5 enzymatic activity. Administration of either MTA or a small-molecule PRMT5 inhibitor showed a modest preferential impairment of cell viability for MTAP-null cancer cell lines compared with isogenic MTAP-expressing counterparts. Together, our findings reveal PRMT5 as a potential vulnerability across multiple cancer lineages augmented by a common "passenger" genomic alteration.


Downregulation of the evolutionary capacitor Hsp90 is mediated by social cues.
The relationship between robustness and evolvability is a long-standing question in evolution. Heat shock protein 90 (HSP90), a molecular chaperone, has been identified as a potential capacitor for evolution, since it allows for the accumulation and release of cryptic genetic variation, and also for the regulation of novel genetic variation through transposon activity. However, to date, it is unknown whether Hsp90 expression is regulated upon demand (i.e. when the release of cryptic genetic variation is most needed). Here, we show that Hsp90 has reduced transcription under conditions where the mobilization of genetic variation could be advantageous. We designed a situation that indicates a stressful environment but avoids the direct effects of stress, by placing untreated (focal) red flour beetles, Tribolium castaneum, into groups together with wounded conspecifics, and found a consistent reduction in expression of two Hsp90 genes (Hsp83 and Hsp90) in focal beetles. We moreover observed a social transfer of immunity in this non-eusocial insect: there was increased activity of the phenoloxidase enzyme and downregulation of the immune regulator, imd. Our study poses the exciting question of whether evolvability might be regulated through the use of information derived from the social environment.


Determination of gallbladder bile lithogenicity in patients with primary biliary cirrhosis.
Patients with primary biliary cirrhosis have a higher prevalence of gallstone disease. Aim of this study was to determine whether gallbladder bile of these patients is lithogenic. We studied 11 patients with early stage primary biliary cirrhosis, and compared them with 16 control subjects. We combined a cholescintigraphic method with nasoduodenal bile sampling to measure the mass of lipids within the gallbladder. Cholesterol saturation index, as measured by standard techniques, was similar in patients with primary biliary cirrhosis and controls (medians: 0.85 vs 0.90). Primary biliary cirrhosis patients showed a significant reduction in the masses of cholesterol, phospholipids and bile acids, as well as in percent biliary deoxycholic acid, as measured by high pressure liquid chromatography (medians 8.6% vs 17.4% in controls; p < 0.05). Percent deoxycholic acid directly correlated with cholesterol mass in all subjects (r = 0.48; p < 0.05). Biliary lipid coupling were similar in the two groups. We conclude that, in patients with early stage primary biliary cirrhosis, gallbladder bile is not lithogenic and biliary lipid coupling is normal, due to a parallel reduction in the masses of cholesterol, phospholipids and bile acids. The significant reduction in percent deoxycholic acid, characteristic of cholestasis, may help explain this biliary lipid mass pattern, that differs from that of cholesterol gallstone patients.


COPD Guidelines: A Review of the 2018 GOLD Report.
Global Strategy for the Diagnosis, Management, and Prevention of COPD 2018 is a consensus report published periodically since 2001 by an international panel of health professionals from respiratory medicine, socioeconomics, public health, and education comprising the Global Initiative for Chronic Obstructive Lung Disease (GOLD). The GOLD documents endeavor to incorporate latest evidence and expert consensus and are intended for use as "strategy documents" for implementation of effective care for chronic obstructive lung disease (COPD) on a global level. The GOLD 2018 report defines COPD as a "common, preventable and treatable disease that is characterized by persistent respiratory symptoms and airflow limitation that is due to airway and/or alveolar abnormalities, usually caused by significant exposure to noxious particles or gases," with the criteria of "persistent respiratory symptoms" being a new and controversial inclusion since 2017. With the availability of newer pharmacotherapy options, treatment recommendations are made on the basis of a review of the latest literature and directed by symptom burden and health care utilization. Apart from the change in definition, a major shift in the recommendations is the exclusion of severity of airflow limitation as one of the major factors in guiding therapy. We review the salient features of the GOLD 2018 document and provide commentary on features that merit further discussion based on our clinical experience and practice as well as literature review current as of February 2018.


HIV Tat-mediated transcriptional regulation of proteasome protein cleavage specificity.
The major antigen-adapted immune response protecting a vertebrate against virus infection is that mediated by CTLs (cytotoxic T-lymphocytes). CTLs destroy virus-infected cells, thereby containing the infection. They are activated by recognition of peptide antigens or epitopes, presented to them in the context of MHC I proteins. These epitopes are derived from proteolytic degradation of endogenously synthesized proteins, which is mediated by the proteasome. Augmentation of epitope presentation by MHC I is thought to be effected by the immunoproteasome, induced in response to IFN-gamma (interferon-gamma) in some cells, and constitutively expressed in others. In this issue of the Biochemical Journal, Remoli and colleagues describe the manipulation of the immunoproteasome by the Tat (transcriptional activation) protein of HIV. The authors show that Tat deregulates the balance of the three proteins, LMP2 (low-molecular-mass polypeptide 2), LMP7 and MECL1 (multicatalytic endopeptidase complex-like 1), which distinguish the immunoproteasome from the proteasome, and they provide a molecular explanation. Intracellular Tat sequesters IRF-1 (interferon-regulatory factor-1) from its cognate promoter element, where normally it associates with STAT1 (signal transducer and activator of transcription 1) to activate basal transcription of the LMP2 gene. LMP2 expression is inhibited as a consequence, skewing the stoichiometry of the immunoproteasome and changing its enzymatic activity. These findings provide a molecular account of an immunomodulatory activity of HIV: changing the peptide antigen profile of cells expressing or exposed to Tat. They may also provide an avenue for manipulating vaccine efficacy and specificity with Tat-based adjuvants.


Vestibular nerve section versus intratympanic gentamicin for Meniere's disease.
Vestibular nerve section and transtympanic gentamicin administration are procedures with proven efficacy in the treatment of vertigo associated with Meniere's disease refractory to medical management. Hearing loss is a known complication of each of these procedures; however, there has not been a report of hearing results of both treatments from a single institution. Retrospective review. Review was made of 25 patients undergoing gentamicin injection and 39 patients undergoing vestibular nerve section for Meniere's disease. Rate of vertigo control and pretreatment and post-treatment pure-tone average values and speech discrimination scores were reported. The mean preoperative pure-tone average for patients having vestibular nerve section was 47.2 dB, with a speech discrimination score of 75.4%. In these patients, the postoperative pure-tone average was 49.1 dB and the speech discrimination score was 75%. Patients undergoing gentamicin injection had a mean pretreatment pure-tone average of 55.9 dB and a speech discrimination score of 62%. The post-treatment pure-tone average and speech discrimination score for the gentamicin group were 68.8 dB and 49.3%, respectively. Five of 25 patients (20%) in the gentamicin treatment group and 1 of 39 (3%) in the vestibular nerve section treatment group had an increase in bone-conduction threshold greater than 30 dB. The amount of postprocedure hearing loss was significantly greater in the gentamicin treatment group (P =.006). Control of vertigo was good to excellent in 95% of the patients treated with vestibular nerve section and in 80% of the patients treated with gentamicin. Although vestibular nerve section and transtympanic gentamicin are both acceptable treatment options for vertigo associated with Meniere's disease, gentamicin causes a higher level of hearing loss related to treatment and vestibular nerve section has higher vertigo control rates.


An evidence-based review of Dopamine receptor agonists in the treatment of Parkinson`s disease.
Apomorphine and certain ergot alkaloids (bromocriptine, lisuride and pergolide) have been available for several decades; for the last few years, they were joined by newer dopamine agonists (cabergoline, pramipexole and ropinirole) most of them are non-ergolines. Each of these dopamine agonists has its own pharmacological characteristics and occupies a place in the pharmacotherapy of Parkinsons disease. In this evidence-based review, emphasis is put on the clinical efficacy of dopamine agonists in early and advanced Parkinsons disease, and where possible comparative evidence regarding their efficacy and safety is provided. In addition, their clinical pharmacokinetics, adverse effect profiles and most relevant interactions will be summarized.


Effect of Motives for Food Choice on Oral Health among Primary School Children in Mangalore: An Analytical Survey.
Parents influence children's eating behaviours by making some foods available than others and by acting as models of eating behaviour. Food selected by parents influence general and oral health of their children. Aim of this study was to assess oral health parameters among primary school children and motives for food choice among their parents in Mangalore. A total of 759 primary school children aged 5-10 years, and their parents participated in this study. Motives for food choice among parents of children were evaluated by using Food Choice Questionnaire (FCQ). Oral health status of students was assessed by using World Health Organisation (WHO) Basic Oral Health Assessment Form. Data pertaining to dietary habits and demographics was also collected. Descriptive and inferential statistics along with Pearson's correlation and Binary logistic regression were executed for the present study and level of significance was fixed at p<0.05. Caries prevalence was 10.8% and 68.9% in permanent and primary dentitions, respectively. Mean Decayed, Missing And Filled Tooth (DMFT) index scores among study subjects were 0.21±0.72 and 3.08±3.11 for permanent and primary dentitions, respectively. Increase in caries prevalence was significantly associated with decrease in total FCQ scores. FCQ domains 'health', 'natural content' and 'weight control' were significantly correlated with dental caries experience. FCQ also showed significant correlation with socio-economic variables. Overall, dental caries experience was found to be high. Parental food choice motives positively influenced dietary patterns and caries experience of their children. Caries experience was less in children whose parents reported higher scores on FCQ. Understanding the barriers, identification of risk factors for poor food choices and targeting interventions might formulate ways by which the desired behaviour can be achieved.


Spontaneous calcification of arteries and cartilage in mice lacking matrix GLA protein.
Calcification of the extracellular matrix (ECM) can be physiological or pathological. Physiological calcification occurs in bone when the soft ECM is converted into a rigid material capable of sustaining mechanical force; pathological calcification can occur in arteries and cartilage and other soft tissues. No molecular determinant regulating ECM calcification has yet been identified. A candidate molecule is matrix GLA protein (Mgp), a mineral-binding ECM protein synthesized by vascular smooth-muscle cells and chondrocytes, two cell types that produce an uncalcified ECM. Mice that lack Mgp develop to term but die within two months as a result of arterial calcification which leads to blood-vessel rupture. Chondrocytes that elaborate a typical cartilage matrix can be seen in the affected arteries. Mgp-deficient mice additionally exhibit inappropriate calcification of various cartilages, including the growth plate, which eventually leads to short stature, osteopenia and fractures. These results indicate that ECM calcification must be actively inhibited in soft tissues. To our knowledge, Mgp is the first inhibitor of calcification of arteries and cartilage to be characterized in vivo.


An introduction to Ayurveda.
Ayurveda is a Sanskrit word derived from two roots: ayur, which means life, and veda, knowledge. Knowledge arranged systematically with logic becomes science. During the due course of time, Ayurveda became the science of life. It has its root in ancient vedic literature and encompasses our entire life, the body, mind, and spirit.


Internal fixation of femoral shaft fractures in children by intramedullary Kirschner wires (a prospective study): its significance for developing countries.
To evaluate internal fixation by intramedullary Kirschner wires as a surgical technique in the treatment of femoral shaft fractures in children by a prospective study. 17 femoral shaft fractures at various levels in 16 children aged 2-15 years were treated by closed intramedullary Kirschner wiring under image intensifier control between May 2000 and October 2003. No external splint was used. Fracture union was achieved in 6-14 weeks. Non-weight bearing crutch walking was started 2-3 days after surgery. Full weight bearing started 6-14 weeks. Average operative time was 40 min (range 20-72 min). Wires were removed after 8-22 weeks. There were no infections, no limb length disparity. One child had pin track ulceration. A big child of 14 years had angulation of the fracture. Intramedullary nailing of femoral shaft fractures in children by stainless steel Kirschner wires is an effective method, which compares well with other studies. It is a simple procedure, which can be easily reproduced. Blood loss is minimal, and the operative time short. There is no need pre-bend the wires in a C or S curve. Stainless steel Kirschner wires are cheap, universally available, and can be manufactured locally. The cost of Image intensifiers is affordable in most of the cities of the developing countries. The hospital does not have to maintain a costly inventory. Provides early mobility, return to home and, school. Gives a predictable clinical pathway and reduces occupancy of hospital beds. The technique was successfully applied for internal fixation of other diaphyseal fractures in children and some selected diaphyseal fractures in adults. Based on my experience and a review of the literature, I recommend this technique as a modality for treatment of femoral shaft fractures in children aged 2 to 14 years.


Aggrecanases and cartilage matrix degradation.
The loss of extracellular matrix macromolecules from the cartilage results in serious impairment of joint function. Metalloproteinases called 'aggrecanases' that cleave the Glu373-Ala374 bond of the aggrecan core protein play a key role in the early stages of cartilage destruction in rheumatoid arthritis and in osteoarthritis. Three members of the ADAMTS family of proteinases, ADAMTS-1, ADAMTS-4 and ADAMTS-5, have been identified as aggrecanases. Matrix metalloproteinases, which are also found in arthritic joints, cleave aggrecans, but at a distinct site from the aggrecanases (i.e. Asn341-Phe342). The present review discuss the enzymatic properties of the three known aggrecanases, the regulation of their activities, and their role in cartilage matrix breakdown during the development of arthritis in relation to the action of matrix metalloproteinases.


Tracing the psychopathology of bipolar disorder to the functional architecture of intrinsic brain activity and its neurotransmitter modulation: a three-dimensional model.
Bipolar disorder (BD) shows complex alterations in psychomotor, affective, and thought dimensions, as described by Kraepelin in his fundamental model of manic-depressive illness. In turn, the expression of behavioral/phenomenological dimensions is traceable to intrinsic brain activity. We reported a data overview on intrinsic brain functioning and its changes in BD. Accordingly, we proposed a three-dimensional model of the relationship between brain functioning and behavioral/phenomenological patterns, along with its application to BD. In this model, intrinsic brain activity is organized in distinct units in accordance to connectivity patterns and related setting of input/output processing, underlying the different behavioral/phenomenological dimensions. An external unit (mainly involving the sensorimotor network) is connected with the external environment and sets the exteroceptive input/somatomotor output processing, underlying the psychomotor dimension. An internal unit (mainly involving the salience network) is connected to the internal/body environment and sets the interoceptive input/visceromotor output processing, underlying the affective dimension. Finally, an associative unit (mainly involving the default-mode network) is not connected with the environment and sets the processing of associative inputs/outputs, underlying the thought dimension. In each unit, neurotransmitter signaling couples the subcortical-cortical loop, which modulates the network activity levels, in turn setting input/output processing and related expression levels of the behavioral/phenomenological dimension. Different combinations in neurotransmitter signaling favor network balancing into distinct functional brain states, which manifest in different combinations of excitation or inhibition in psychomotricity, affectivity, and thought, resulting in the manic, depressive, and mixed states of BD. Our working model might provide a coherent framework for tracing the complex BD psychopathology to core functional brain alterations.


Sequences responsible for intracellular localization of beta-actin messenger RNA also affect cell phenotype.
We have characterized the structure and function of RNA sequences that direct beta-cytoplasmic actin mRNA to the cell periphery were mapped to two segments of 3'-untranslated region by expression of LacZ/beta-actin chimeric mRNAs in chicken embryo fibroblasts (CEFs). A 54-nt segment, the "RNA zipcode," and a homologous but less active 43-nt segment each localized beta-galactosidase activity to the leading lamellae. This zipcode contains the full activity, and mutations or deletions within it reduce, but do not eliminate, its activity, indicating that several motifs contribute to the activity. Two of these motifs, when multimerized, can regenerate almost full activity. These sequences are highly conserved in evolution, since the human beta-actin zipcode, positioned identically in the 3'UTR localizes equally well in chicken cells. Complementary phosphorothioate oligonucleotides against the zipcode delocalized endogenous beta-actin mRNA, whereas those complementary to the region just outside the zipcode, or sense oligonucleotides, did not. Actin mRNA or protein levels were unaffected by the antisense treatments, but a dramatic change in lamellipodia structure, and actin stress fiber organization was observed using the same antizipcode oligonucleotides which delocalized the mRNA. Hence, discrete 3'UTR sequences direct beta-actin isoform synthesis to the leading lamellae and affect cell morphology, presumably through the actin cytoskeleton.


Validation and noninvasive kinetic modeling of [11C]UCB-J PET imaging in mice.
Synaptic pathology is associated with several brain disorders, thus positron emission tomography (PET) imaging of synaptic vesicle glycoprotein 2A (SV2A) using the radioligand [11C]UCB-J may provide a tool to measure synaptic alterations. Given the pivotal role of mouse models in understanding neuropsychiatric and neurodegenerative disorders, this study aims to validate and characterize [11C]UCB-J in mice. We performed a blocking study to verify the specificity of the radiotracer to SV2A, examined kinetic models using an image-derived input function (IDIF) for quantification of the radiotracer, and investigated the in vivo metabolism. Regional TACs during baseline showed rapid uptake of [11C]UCB-J into the brain. Pretreatment with levetiracetam confirmed target engagement in a dose-dependent manner. VT (IDIF) values estimated with one- and two-tissue compartmental models (1TCM and 2TCM) were highly comparable (r=0.999, p < 0.0001), with 1TCM performing better than 2TCM for K1 (IDIF). A scan duration of 60 min was sufficient for reliable VT (IDIF) and K1 (IDIF) estimations. In vivo metabolism of [11C]UCB-J was relatively rapid, with a parent fraction of 22.5 ± 4.2% at 15 min p.i. In conclusion, our findings show that [11C]UCB-J selectively binds to SV2A with optimal kinetics in the mouse representing a promising tool to noninvasively quantify synaptic density in comparative or therapeutic studies in neuropsychiatric and neurodegenerative disorder models.


In silico characterization and prediction of global protein-mRNA interactions in yeast.
Post-transcriptional gene regulation is mediated through complex networks of protein-RNA interactions. The targets of only a few RNA binding proteins (RBPs) are known, even in the well-characterized budding yeast. In silico prediction of protein-RNA interactions is therefore useful to guide experiments and to provide insight into regulatory networks. Computational approaches have identified RBP targets based on sequence binding preferences. We investigate here to what extent RBP-RNA interactions can be predicted based on RBP and mRNA features other than sequence motifs. We analyze global relationships between gene and protein properties in general and between selected RBPs and known mRNA targets in particular. Highly translated RBPs tend to bind to shorter transcripts, and transcripts bound by the same RBP show high expression correlation across different biological conditions. Surprisingly, a given RBP preferentially binds to mRNAs that encode interaction partners for this RBP, suggesting coordinated post-transcriptional auto-regulation of protein complexes. We apply a machine-learning approach to predict specific RBP targets in yeast. Although this approach performs well for RBPs with known targets, predictions for uncharacterized RBPs remain challenging due to limiting experimental data. We also predict targets of fission yeast RBPs, indicating that the suggested framework could be applied to other species once more experimental data are available.


Regional needs assessment for emergency physician audit and feedback.
Audit and feedback is widely used to improve physician performance. Many data metrics are being provided to physicians, yet most of these are driven by the regulatory environment. We sought to conduct a needs assessment of audit and feedback metrics that were most useful to clinicians within our health care region. We conducted a Web-based survey of five clinical practice sites in our region and asked that physicians rank 49 clinical practice metrics. In addition, we assessed their readiness for audit and feedback and their preferences for data confidentiality. We collected data on duration of training, gender, and site of practice (academic v. community) allowing for comparison between groups. A total of 104 emergency medicine physicians participated in the survey (52.3% response rate). There was a significant readiness for participation in audit and feedback activities. Top ranked metrics were emergency department return rates and colleague's assessment of collegiality and quality of care, which were common across all sites. Small yet significant differences were noted between genders and academic v. community practitioners. This study represents the first regional analysis of physician preferences for audit and feedback activities and implementation. It demonstrates that physicians are interested in audit and feedback activities and provides a roadmap for the development of a regional audit and feedback structure. It will also be used as a guiding document for regional change management.


Leukocyte analysis from WHIM syndrome patients reveals a pivotal role for GRK3 in CXCR4 signaling.
Leukocytes from individuals with warts, hypogammaglobulinemia, infections, and myelokathexis (WHIM) syndrome, a rare immunodeficiency, and bearing a wild-type CXCR4 ORF (WHIM(WT)) display impaired CXCR4 internalization and desensitization upon exposure to CXCL12. The resulting enhanced CXCR4-dependent responses, including chemotaxis, probably impair leukocyte trafficking and account for the immunohematologic clinical manifestations of WHIM syndrome. We provided here evidence that GPCR kinase-3 (GRK3) specifically regulates CXCL12-promoted internalization and desensitization of CXCR4. GRK3-silenced control cells displayed altered CXCR4 attenuation and enhanced chemotaxis, as did WHIM(WT) cells. These findings identified GRK3 as a negative regulator of CXCL12-induced chemotaxis and as a candidate responsible for CXCR4 dysfunction in WHIM(WT) leukocytes. Consistent with this, we showed that GRK3 overexpression in both leukocytes and skin fibroblasts from 2 unrelated WHIM(WT) patients restored CXCL12-induced internalization and desensitization of CXCR4 and normalized chemotaxis. Moreover, we found in cells derived from one patient a profound and selective decrease in GRK3 products that probably resulted from defective mRNA synthesis. Taken together, these results have revealed a pivotal role for GRK3 in regulating CXCR4 attenuation and have provided a mechanistic link between the GRK3 pathway and the CXCR4-related WHIM(WT) disorder.


Resitting a high-stakes postgraduate medical examination on multiple occasions: nonlinear multilevel modelling of performance in the MRCP(UK) examinations.
Failure rates in postgraduate examinations are often high and many candidates therefore retake examinations on several or even many times. Little, however, is known about how candidates perform across those multiple attempts. A key theoretical question to be resolved is whether candidates pass at a resit because they have got better, having acquired more knowledge or skills, or whether they have got lucky, chance helping them to get over the pass mark. In the UK, the issue of resits has become of particular interest since the General Medical Council issued a consultation and is considering limiting the number of attempts candidates may make at examinations. Since 1999 the examination for Membership of the Royal Colleges of Physicians of the United Kingdom (MRCP(UK)) has imposed no limit on the number of attempts candidates can make at its Part 1, Part 2 or PACES (Clinical) examination. The present study examined the performance of candidates on the examinations from 2002/2003 to 2010, during which time the examination structure has been stable. Data were available for 70,856 attempts at Part 1 by 39,335 candidates, 37,654 attempts at Part 2 by 23,637 candidates and 40,303 attempts at PACES by 21,270 candidates, with the maximum number of attempts being 26, 21 and 14, respectively. The results were analyzed using multilevel modelling, fitting negative exponential growth curves to individual candidate performance. The number of candidates taking the assessment falls exponentially at each attempt. Performance improves across attempts, with evidence in the Part 1 examination that candidates are still improving up to the tenth attempt, with a similar improvement up to the fourth attempt in Part 2 and the sixth attempt at PACES. Random effects modelling shows that candidates begin at a starting level, with performance increasing by a smaller amount at each attempt, with evidence of a maximum, asymptotic level for candidates, and candidates showing variation in starting level, rate of improvement and maximum level. Modelling longitudinal performance across the three diets (sittings) shows that the starting level at Part 1 predicts starting level at both Part 2 and PACES, and the rate of improvement at Part 1 also predicts the starting level at Part 2 and PACES. Candidates continue to show evidence of true improvement in performance up to at least the tenth attempt at MRCP(UK) Part 1, although there are individual differences in the starting level, the rate of improvement and the maximum level that can be achieved. Such findings provide little support for arguments that candidates should only be allowed a fixed number of attempts at an examination. However, unlimited numbers of attempts are also difficult to justify because of the inevitable and ever increasing role that luck must play with increasing numbers of resits, so that the issue of multiple attempts might be better addressed by tackling the difficult question of how a pass mark should increase with each attempt at an exam.


Deep water masses and sediments are main compartments for polychlorinated biphenyls in the Arctic Ocean.
There is a wealth of studies of polychlorinated biphenyls (PCB) in surface water and biota of the Arctic Ocean. Still, there are no observation-based assessments of PCB distribution and inventories in and between the major Arctic Ocean compartments. Here, the first water column distribution of PCBs in the central Arctic Ocean basins (Nansen, Amundsen, and Makarov) is presented, demonstrating nutrient-like vertical profiles with 5-10 times higher concentrations in the intermediate and deep water masses than in surface waters. The consistent vertical profiles in all three Arctic Ocean basins likely reflect buildup of PCBs transported from the shelf seas and from dissolution and/or mineralization of settling particles. Combined with measurement data on PCBs in other Arctic Ocean compartments collected over the past decade, the total Arctic Ocean inventory of ∑7PCB was estimated to 182 ± 40 t (±1 standard error of the mean), with sediments (144 ± 40 t), intermediate (5 ± 1 t) and deep water masses (30 ± 2 t) storing 98% of the PCBs in the Arctic Ocean. Further, we used hydrographic and carbon cycle parametrizations to assess the main pathways of PCBs into and out of the Arctic Ocean during the 20th century. River discharge appeared to be the major pathway for PCBs into the Arctic Ocean with 115 ± 11 t, followed by ocean currents (52 ± 17 t) and net atmospheric deposition (30 ± 28 t). Ocean currents provided the only important pathway out of the Arctic Ocean, with an estimated cumulative flux of 22 ± 10 t. The observation-based inventory of ∑7PCB of 182 ± 40 t is consistent with the contemporary inventory based on cumulative fluxes for ∑7PCB of 173 ± 36 t. Information on the concentration and distribution of PCBs in the deeper compartments of the Arctic Ocean improves our understanding of the large-scale fate of POPs in the Arctic and may also provide a means to test and improve models used to assess the fate of organic pollutants in the Arctic.


Deaths from varicella in infants.
Onset of maternal varicella up to 5 days before delivery is associated with in utero exposure to varicella and may result in severe infection in the newborn 5 to 10 days after delivery. Since up to 31% of these newborns may die, routine administration of varicella-zoster immunoglobulin to these infants is recommended. Little is known, however, about the risk of death in other infants with postnatal infection. Available epidemiologic data indicate an estimated death/case ratio for children less than 1 year of age 4 times that for 1- to 14-year-olds (8 in 100,000 vs. 2 in 100,000). Since the actual ages for the infant deaths were lacking, it has been impossible to know how many deaths were possibly related to maternal varicella contracted within the 5 days before delivery. Using National Center for Health Statistics data, we analyzed 92 deaths due to varicella in children less than 1 year old reported between 1968 and 1978 (median age, 5.5 months). Only five deaths occurred in newborns (ages 8 hours to 19 days). These data indicate that intrauterine infection accounts for few varicella deaths in infants. Since postnatal infection accounts for the observed increased risk of death in this age group, the need for preventing postnatal varicella in all infants merits further study. However, based on the small number of deaths occurring annually and the low relative risk compared to other high risk groups, routine postexposure administration of varicella-zoster immunoglobulin to all children less than 1 year of age does not seem warranted at this time.


The genomic signature of human rhinoviruses A, B and C.
Human rhinoviruses are single stranded positive sense RNA viruses that are presented in more than 50% of acute upper respiratory tract infections. Despite extensive studies on the genetic diversity of the virus, little is known about the forces driving it. In order to explain this diversity, many research groups have focused on protein sequence requirements for viable, functional and transmissible virus but have missed out an important aspect of viral evolution such as the genomic ontology of the virus. This study presents for the first time the genomic signature of 111 fully sequenced HRV strains from all three groups HRV-A, HRV-B and HRV-C. We observed an HRV genome tendency to eliminate CpG and UpA dinucleotides, coupling with over-representation of UpG and CpA. We propose a specific mechanism which describes how rapid changes in the HRV genomic sequence can take place under the strict control of conservation of the polypeptide backbone. Moreover, the distribution of the observed under- and over-represented dinucleotides along the HRV genome is presented. Distance matrice tables based on CpG and UpA odds ratios were constructed and viewed as heatmaps and distance trees. None of the suppressions can be attributed to codon usage or in RNA secondary structure requirements. Since viral recognition is dependent on RNA motifs rich in CpG and UpA, it is possible that the overall described genome evolution mechanism acts in order to protect the virus from host recognition.


Delivery of the autofluorescent protein R-phycoerythrin by calcium phosphate nanoparticles into four different eukaryotic cell lines (HeLa, HEK293T, MG-63, MC3T3): Highly efficient, but leading to endolysosomal proteolysis in HeLa and MC3T3 cells.
Nanoparticles can be used as carriers to transport biomolecules like proteins and synthetic molecules across the cell membrane because many molecules are not able to cross the cell membrane on their own. The uptake of nanoparticles together with their cargo typically occurs via endocytosis, raising concerns about the possible degradation of the cargo in the endolysosomal system. As the tracking of a dye-labelled protein during cellular uptake and processing is not indicative of the presence of the protein itself but only for the fluorescent label, a label-free tracking was performed with the red-fluorescing model protein R-phycoerythrin (R-PE). Four different eukaryotic cell lines were investigated: HeLa, HEK293T, MG-63, and MC3T3. Alone, the protein was not taken up by any cell line; only with the help of calcium phosphate nanoparticles, an efficient uptake occurred. After the uptake into HeLa cells, the protein was found in early endosomes (shown by the marker EEA1) and lysosomes (shown by the marker Lamp1). There, it was still intact and functional (i.e. properly folded) as its red fluorescence was detected. However, a few hours after the uptake, proteolysis started as indicated by the decreasing red fluorescence intensity in the case of HeLa and MC3T3 cells. 12 h after the uptake, the protein was almost completely degraded in HeLa cells and MC3T3 cells. In HEK293T cells and MG-63 cells, no degradation of the protein was observed. In the presence of Bafilomycin A1, an inhibitor of acidification and protein degradation in lysosomes, the fluorescence of R-PE remained intact over the whole observation period in the four cell lines. These results indicate that despite an efficient nanoparticle-mediated uptake of proteins by cells, a rapid endolysosomal degradation may prevent the desired (e.g. therapeutic) effect of a protein inside a cell.


Comparative genomics guided discovery of two missing archaeal enzyme families involved in the biosynthesis of the pterin moiety of tetrahydromethanopterin and tetrahydrofolate.
C-1 carriers are essential cofactors in all domains of life, and in Archaea, these can be derivatives of tetrahydromethanopterin (H(4)-MPT) or tetrahydrofolate (H(4)-folate). Their synthesis requires 6-hydroxymethyl-7,8-dihydropterin diphosphate (6-HMDP) as the precursor, but the nature of pathways that lead to its formation were unknown until the recent discovery of the GTP cyclohydrolase IB/MptA family that catalyzes the first step, the conversion of GTP to dihydroneopterin 2',3'-cyclic phosphate or 7,8-dihydroneopterin triphosphate [El Yacoubi, B.; et al. (2006) J. Biol. Chem., 281, 37586-37593 and Grochowski, L. L.; et al. (2007) Biochemistry46, 6658-6667]. Using a combination of comparative genomics analyses, heterologous complementation tests, and in vitro assays, we show that the archaeal protein families COG2098 and COG1634 specify two of the missing 6-HMDP synthesis enzymes. Members of the COG2098 family catalyze the formation of 6-hydroxymethyl-7,8-dihydropterin from 7,8-dihydroneopterin, while members of the COG1634 family catalyze the formation of 6-HMDP from 6-hydroxymethyl-7,8-dihydropterin. The discovery of these missing genes solves a long-standing mystery and provides novel examples of convergent evolutions where proteins of dissimilar architectures perform the same biochemical function.


Lucidumol D, a new lanostane-type triterpene from fruiting bodies of Reishi (Ganoderma lingzhi).
A new lanostane-type triterpenoid, lucidumol D (1) was isolated from the fruiting bodies of Ganoderma lingzhi. Its structure was elucidated on the basis of extensive 1D- and 2D-NMR studies as well as mass spectrometry. The cytotoxicity of lucidumol D against proliferation of several cancer cells were assayed by using MTT method and the obtained result suggested selective anti-proliferative and cytotoxic effects against MCF-7, HepG2, HeLa, Caco-2, and HCT-116. In comparison to lucidumol C (2) isolated previously by our group, the structure-activity relationship indicated that carbonyl function at C-11 is necessary to enhance the cytotoxicity.


Risk of Surgery and Mortality in Elderly-onset Inflammatory Bowel Disease: A Population-based Cohort Study.
As the prevalence of inflammatory bowel disease (IBD) increases in the elderly population, we sought to characterize IBD-related outcomes in this population. We identified incident IBD cases in Ontario, Canada between 1999 and 2008 and categorized subjects by age at diagnosis as young adults (18-40 yr); middle-age adults (41-64 yr); and elderly (≥65 yr) from within population-based health administrative data. We determined the risk of IBD-related surgery and mortality in those with elderly-onset IBD compared with other age groups. Of 21,218 persons with IBD, there were 1749 cases of elderly-onset ulcerative colitis (UC) and 725 cases elderly-onset Crohn's disease (CD). Elderly UC had higher rates of IBD-related surgery than those with young-adult UC (adjusted hazard ratio, 1.34; 95% CI, 1.16-1.55), although there was no difference in surgical rates between age groups in CD. IBD-specific mortality was higher in elderly-onset CD (33.1/10,000 person-year) compared with that in middle-age CD (5.6/10,000 person-year, P < 0.0001) and young adult CD (1.0/10,000 person-year) but was not different by age in UC. The leading cause of death in elderly UC and CD was solid malignancies accounting for 22.9% and 26.4% of deaths, respectively, whereas IBD was third most frequent cause of death accounting for 6.3% and 9.1% of deaths, respectively. Elderly-onset patients with UC were more likely to undergo surgery while elderly-onset patients with CD exhibited higher IBD-specific mortality than those with younger-onset disease. These findings should prompt more optimized disease management in elderly IBD.